Lymph directing prodrugs

Abstract

To provide compounds in the form of prodrugs that promote transport of a pharmaceutical agent to the lymphatic system and subsequently enhance release of a parent drug.SOLUTION: The invention provides a compound represented by the formula (I) or a pharmaceutically acceptable salt thereof. In the formula: R1 and R2 independently represent H or a residue of a fatty acid; X is selected from among -O-, -NH-, and -S-; -L-A represents a residue of a pharmaceutical agent; -L- is -OC(O)- or -X'-; -Y- represents an -alkyl C(O)OCH2-, -alkenyl C(O)OCH2-, or -alkynyl C(O)OCH2- group when -L- is -OC(O)-, or -Y- represents an -alkyl C(O)R3-, -alkenyl C(O)R3-, -alkynyl C(O)R3- group when -L- is -X'-; R3 is a self-immolative group; X' is O, S, N(R4), or N(H)S(O)2; and R4 is H or alkyl.SELECTED DRAWING: None

Description

Detailed Description of the Invention

[0001] FIELD OF THE INVENTION The present invention relates to prodrug forms of compounds, particularly compounds that facilitate the transport of drugs into the lymphatic system and subsequently enhance the release of the parent drug.

[0002] Background of the Invention The lymphatic system consists of a specialized network of ducts, nodes, and lymphoid tissues distributed throughout the body in close proximity to the vascular system. The lymphatic system plays many important roles in immune responses, fluid balance, nutrient absorption, lipid homeostasis, and tumor metastasis. Due to the unique anatomical and physiological properties of the lymphatic system, targeted delivery of drugs to and through the lymphatic system has been proposed as a means to improve both pharmacokinetic and pharmacodynamic profiles. Lymphatic drug transport promises to enhance oral bioavailability through bypass of first-pass metabolism, alter systemic drug disposition, and enhance efficacy against lymphatic or lymphocyte-mediated pathologies, such as lymphoma, leukemia, lymphoid tumor metastasis, autoimmune diseases, lymph-resident infections, and graft rejection.

[0003] For drugs to access the intestinal lymph, they must first associate with intestinal lymphatic lipoproteins, which are recruited to intestinal absorptive cells (enterocytes) in response to lipid absorption. Association with these lipoproteins facilitates subsequent lymphatic transport. This is because the size of lipoproteins precludes rapid diffusion across the vascular endothelium, which lines the capillaries through which the contents of the small intestine flow. Instead, these large colloidal structures enter lymphatic capillaries, where the lymphatic endothelium is significantly more permeable than the vascular endothelium. Historically, drugs with high lymphatic transport have been highly lipophilic (typically, but not limited to, a logD > 5 and a solubility in long-chain triglycerides > 50 mg / g) to facilitate physical association with lipoproteins. Therefore, highly lipophilic drug analogs have been envisioned as a way to enhance lymphatic transport of drugs. However, chemical modification of the parent drug can result in reduced efficacy, and a significant increase in lipophilicity is often correlated with increased toxicity.

[0004] Lipophilic prodrug forms of compounds provide a means to temporarily increase the lipophilicity and lipoprotein affinity of pharmaceutical compounds, thereby increasing lymphatic targeting. Upon transport through the lymphatic system, the prodrug ultimately reverts to the parent drug so that it becomes active at its target site.

[0005] Several studies have investigated the potential of simple aliphatic esters of drugs as lymphatic-directed prodrugs. Testosterone undecanoate is an example of a commercially available compound that has adopted this approach. After oral administration, testosterone is almost completely metabolized on its first pass through the liver, resulting in minimal bioavailability. Testosterone undecanoate redirects a small percentage of the absorbed dose to the lymphatic system, thereby avoiding first-pass metabolism in the liver and increasing the oral bioavailability of testosterone. However, this process is still very inefficient, and the bioavailability of testosterone after oral administration of the undecanoate ester is believed to be less than 5%.

[0006] Another mechanism for enhancing lymphatic transport of drugs is to employ prodrugs that are incorporated into endogenous pathways associated with the absorption, transport, and disposition of dietary lipids. An example of a dietary lipid that can be utilized is triglyceride. Examples of drug-lipid conjugates in which the parent drug contains an available carboxylic acid group and is directly conjugated to the glyceride backbone have been described in many prior publications (Paris, G. et al., J. Med. Chem. 1979, 22, (6), 683-687; Garzon Aburbeh, A. et al., J. Med. Chem. 1983, 26, (8), 1200-1203; Deverre, J. R. et al., J. Pharm. Pharmacol. 1989, 41, (3), 191-193; Mergen, F. et al., J. Pharm. Pharmacol. 1991, 43, (11), 815-816; Garzon Aburbeh, A. et al., J. Med. Chem. 1986, 29, (5), 687-69; and Han, S. et al. al.J.Control.Release 2014,177,1-10).

[0007] In another example, when a drug does not contain an available carboxylic acid, a short linker has been used to facilitate drug-triglyceride conjugation (Scriba, GKE, Arch. Pharm. (Weinheim). 1995, 328, (3), 271-276, and Scriba, GKE et al., J. Pharm. Pharmacol. 1995, 47, (11), 945-948). These drug-lipid conjugates employ succinic acid to facilitate conjugation to available hydroxyl functional groups. However, the literature teaches that this structure is not at all useful. For example, Scriba investigated the in vitro hydrolysis of testosterone-succinic acid-glyceride lipid conjugates and concluded that "testosterone was only very slowly released from the prodrug in this study due to chemical hydrolysis catalyzed by plasma esterases and mediated by lipases. Thus, testosterone conjugates appear to be poor prodrugs for steroid delivery."

[0008] Others have employed ether bonds to the glyceride and ester bonds to the drug (Sugihara, J. et al., J. Pharmacobiodyn. 1988, 11, (5), 369-376, and Sugihara, J. et al., J. Pharmacobiodyn. 1988, 11, (8), 555-562). The authors of these papers clearly state that the ether bond between the glycerol and the n-alkyl chain and the ester bond between the n-alkyl chain and the drug appear to be necessary for chemical modification of the drug. However, the present inventors have found that the ether bond is actually unproductive and does not allow significant lymphatic transport.

[0009] Therefore, there is a need to develop novel lipid-drug conjugates that facilitate stable transport of drugs to the intestinal lymph and that are readily reverted to the active parent drug.

[0010] Summary of the Invention It has now been found that the use of a self-immolative group and certain linkers to link the drug to the triglyceride unit provides an optimal pharmacokinetic profile for the resulting lipid-drug conjugate.

[0011] Thus, in one aspect, the present invention provides a compound of formula (I): [ka] [In the formula, R 1 and R 2 are independently H or C2-C 28 represents a residue of a fatty acid, -X- is selected from -O-, -NH-, and -S-; [ka] represents the residue of a drug, -L- is -OC(O)- or -X'-; When -L- is -OC(O)-, -Y- is an optionally substituted -C1 to C 20 Alkyl C(O)OCH2-, -C2~C 20 Alkenyl C(O)OCH2- or -C2-C 20 represents an alkynyl C(O)OCH2- group, in which one or more of the carbon atoms of the alkyl, alkenyl, or alkynyl group may be replaced by NH, S, O, a C5-C8 aromatic or aliphatic cyclic group, or a C5-C8 aromatic or aliphatic heterocyclic group, provided that the alkyl, alkenyl, or alkynyl group is not a straight-chain C 20 provided that the length does not exceed the length of the alkyl group; or When -L- is -X'-, -Y- is an optionally substituted -C1-C2 alkylC(O)R 3 - group, or -C2 alkenyl C(O)R 3 - or -C2 alkynylC(O)R 3represents a - group, R 3 is a self-immolative group, X' is O, S, N(R 4 ), or N(H)S(O)2, R 4 is H or C1-C4 alkyl; or Pharmaceutically acceptable salts thereof are provided.

[0012] In a further aspect, the present invention provides a compound of formula (I) represented by formula (V): [ka] [In the formula, R 1 , R 2 and -X- are as defined for formula (I), R 5 and R 6 are independently selected from hydrogen and C1-C4 alkyl; R 3 is a self-immolative group], or Pharmaceutically acceptable salts thereof are provided.

[0013] In another aspect, the present invention provides a method for treating or preventing a disease or disorder in which elevated testosterone levels are beneficial, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of formula (V) The present invention provides a method comprising administering a compound of formula (I) to a subject.

[0014] In a further aspect, the present invention provides the use of a compound of formula (V) in the manufacture of a medicament for treating or preventing a disease or disorder in which elevated testosterone levels are beneficial.

[0015] In another aspect, the present invention provides a compound of formula (V) for use in the treatment or prevention of a disease or disorder in which elevated testosterone levels would be beneficial.

[0016] In another aspect, the present invention provides a method for enhancing lymphatic transport and systemic release of a drug, comprising providing the drug with a prodrug moiety of formula (VI): [ka] [In the formula, R 1 and R 2 are independently H or C2-C 28 represents a residue of a fatty acid, -X- is selected from -O-, -NH-, and -S-; When -L- is -X'-, -Y- is an optionally substituted -C1-C2 alkylC(O)R 3 - group, or -C2 alkenyl C(O)R 3 - or -C2 alkynylC(O)R 3 represents a - group, R 3 is a self-immolative group, [ka] indicates the point at which the linker is conjugated to the pharmaceutically active agent], or and conjugating a pharmaceutically acceptable salt thereof.

[0017] These and other aspects of the present invention will become more apparent to those skilled in the art upon reading the following detailed description in conjunction with the accompanying examples and claims. [Brief explanation of the drawings]

[0018] [Figure 1] 1 is a graphical representation of the cumulative lymphatic transport (% of dose) of total testosterone-related derivatives versus time in anesthetized, mesenteric lymphatic cannulated female SD rats following intraduodenal infusion of testosterone, testosterone undecanoate (TU), Compound 21, Compound 12, Compound 13 / 14, Compound 15, Compound 17, and Compound 19. [Figure 2]FIG. 1 is a graphical representation of dose-normalized testosterone plasma concentrations following oral gavage administration of testosterone, testosterone undecanoate (TU), Compound 12, Compound 15, Compound 16, Compound 17, Compound 18, Compound 20, and Compound 21 to conscious, carotid-cannulated female SD rats. [Figure 3] FIG. 1 is a graphical representation of dose-normalized testosterone plasma concentrations following oral gavage administration of testosterone, testosterone undecanoate (TU), Compound 12, Compound 13, Compound 13 / 14, Compound 18, and Compound 19 to conscious, carotid-cannulated female SD rats. [Figure 4] 1 is a graphical representation of cumulative lymphatic transport (% of dose) of total SER-related derivatives over time in anesthetized, mesenteric lymphatic cannulated male SD rats following intraduodenal infusion of sertraline hydrochloride (SER.HCl) and Compound 3. [Figure 5] 1 is a graphical representation of dose-normalized SER plasma concentrations following oral gavage administration of sertraline hydrochloride (SER.HCl), Compound 1, Compound 2, and Compound 3. [Figure 6] 1 is a graphical representation of the cumulative lymphatic transport (% of dose) of all buprenorphine (BUP)-related derivatives over time in anesthetized, mesenteric lymphatic cannulated male SD rats following intraduodenal infusion of BUP, Compound 6, and Compound 7. [Figure 7] 1 is a graphical representation of dose-normalized buprenorphine (BUP) plasma concentrations following oral gavage administration of BUP, Compound 5, Compound 6, and Compound 7. [Figure 8] 1 is a graphical representation of the cumulative lymphatic transport (% of dose) of all mycophenolic acid (MPA)-related derivatives over time in anesthetized, mesenteric lymphatic cannulated male SD rats following intraduodenal infusion of MPA, Compound 10, and Compound 11. [Figure 9]1 is a graphical representation of the stability profiles of the monoglyceride forms of Compound 12, Compound 13 / 14, Compound 15, Compound 16, Compound 17, Compound 20, and Compound 21 during in vitro incubation with porcine pancreatic lipase. [Figure 10] 1 is a graphical representation of the release of MPA from 1,3-dipalmitoylglycerol mycophenolate (MPA-TG), Compound 10, and Compound 11.

[0019] Detailed Description of the Invention When prodrug strategies are employed in the field of drug development to improve pharmacokinetic properties, the prodrug is usually expected to revert to the parent compound via nonspecific degradation or enzyme-mediated biotransformation before exhibiting biological activity. The present invention discloses modified glyceride-based compounds that can facilitate lymphatic transport of drugs and improve the reversion of the compound to the active drug.

[0020] Dietary lipids, such as triglycerides, use a unique metabolic pathway to gain access to the lymph (and ultimately the systemic circulation) that is completely separate from other nutrients, such as proteins and carbohydrates. After ingestion, dietary triglycerides are hydrolyzed by luminal lipases to release one monoglyceride and two fatty acids for each triglyceride molecule. The monoglyceride and two fatty acids are then absorbed into enterocytes, where they are re-esterified to triglycerides.

[0021] Re-synthesized triglycerides are assembled into intestinal lipoproteins (primarily chylomicrons), which gain preferential access to the intestinal lymphatics after exocytosis from enterocytes. Within the lymphatics, lipids in the form of chylomicrons exit a series of capillaries, nodes, and ducts, all of which empty into the systemic circulation at the junction of the left subclavian and internal jugular veins. After entering the circulation, triglycerides as chylomicrons are preferentially and efficiently taken up by tissues with high expression of lipoprotein lipase, such as adipose tissue, the liver, and potentially certain types of tumor tissue.

[0022] Lipid mimetic compounds behave similarly to natural triglycerides and are expected to be transported to and through the lymphatic system before reaching the systemic circulation. In this way, the pharmacokinetic and pharmacodynamic profiles of the parent drug can be manipulated to enhance access to lymph and lymphoid tissues, thereby circumventing first-pass metabolism (and potentially intestinal outflow), thereby promoting oral bioavailability. Lipid mimetic compounds also promote delivery to sites within lymph, lymph nodes, and lymphoid tissues, as well as to sites of high lipid utilization and lipoprotein lipase expression, such as adipose tissue, liver, and some tumors. Drug targeting can also be facilitated.

[0023] Lipidated prodrugs, which are readily converted to the parent drug after transport through the systemic circulation, can reduce free drug concentrations in the gastrointestinal (GI) tract, thereby providing benefits in reducing gastrointestinal irritation, taste masking, facilitating drug solubilization as intestinal bile salt micelles (due to similarity to endogenous monoglycerides), and enhancing passive membrane permeability (by increasing lipophilicity). Lipidated prodrugs also enhance solubility in lipid vehicles, including lipids alone or mixtures of lipids with surfactants and / or cosolvents, allowing larger doses of the drug to be administered in solution than would be possible with the parent drug.

[0024] The present inventors have surprisingly found that the portion of a drug-glyceride conjugate that links the drug to the glyceride unit can be modified to improve the stability of the drug-glyceride conjugate in the GI tract, promote transport to the intestinal lymph, and ultimately promote the release of the drug from the drug-glyceride prodrug. Thus, by altering the "linker" that links the drug to the glyceride unit, an optimal pharmacokinetic profile can be achieved for the resulting compound.

[0025] The present inventors have found that incorporating a self-immolative group into the linker between the drug and the glyceride unit results in improved systemic release and exposure of the drug, even when the linker is a short-chain linker (i.e., succinic acid as previously reported). The incorporation of a self-immolative group enhances the release of the drug in the systemic circulation. Although the self-immolative group may reduce lymphatic transport of the drug, it has been found for the first time that even if lymphatic transport is reduced, systemic drug exposure is enhanced, likely due to enhanced drug release in the systemic circulation. Furthermore, it has been found that employing methyl substitutions on the carbon atoms of the linker between the self-immolative group and the glyceride unit can enhance stability in the GI tract, enhance lymphatic transport, and still maintain good conversion in the systemic circulation.

[0026] A number of terms are used herein that are well known to those of ordinary skill in the art. Nevertheless, for the sake of clarity, a number of terms are defined.

[0027] As used herein, unless otherwise defined, the term "optionally substituted" is intended to mean that a group may be unsubstituted or further substituted with one or more groups selected from hydroxyl, alkyl, alkoxy, alkoxycarbonyl, alkenyl, alkenyloxy, alkynyl, alkynyloxy, amino, aminoacyl, thio, arylalkyl, arylalkoxy, aryl, aryloxy, acylamino, carboxy, cyano, halogen, nitro, sulfo, phosphono, phosphorylamino, phosphinyl, heteroaryl, heteroaryloxy, heterocyclyl, heterocyclooxy, trihalomethyl, pentafluoroethyl, trifluoromethoxy, difluoromethoxy, trifluoromethanethio, trifluoroethenyl, mono- and di-alkylamino, mono- and di-(substituted alkyl)amino, mono- and di-arylamino, mono- and di-heteroarylamino, mono- and di-heterocyclyl, amino, and asymmetric disubstituted amines having different substituents selected from alkyl, aryl, heteroaryl, and heterocyclyl.

[0028] As used herein, the term "alkyl" used alone or in compound words refers to a straight chain alkyl or a branched chain alkyl. 20 " is used to indicate the number of carbon atoms in the alkyl group (in this case, 2 to 20). Examples of straight and branched chain alkyls are methyl, ethyl, n-propyl, isopropyl, and methyl. Propyl, n-butyl, sec-butyl, t-butyl, n-pentyl, hexyl, heptyl, 5-methylheptyl, 5-methylhexyl, octyl, nonyl, decyl, undecyl, dodecyl, and docosyl (C 22 ) are mentioned.

[0029] As used herein, the term "alkenyl," used alone or in compound terms, refers to a straight-chain or branched-chain hydrocarbon residue containing at least one carbon-to-carbon double bond, including ethylenically mono-, di-, or polyunsaturated alkyl, as defined above. Preferably, the alkenyl group is a straight-chain alkenyl group. "C2-C 20 " is used to indicate the number of carbon atoms in the alkenyl group (in this case, 2 to 20). Examples of alkenyl include vinyl, allyl, 1-methylvinyl, butenyl, iso-butenyl, 3-methyl-2-butenyl, 1-pentenyl, 1-hexenyl, 3-hexenyl, 1-heptenyl, 3-heptenyl, 1-octenyl, 1-nonenyl, 2-nonenyl, 3-nonenyl, 1-decenyl, 3-decenyl, 1,3-butadienyl, 1,4-pentadienyl, 1,3-hexadienyl, 1,4-hexadienyl, and 5-docosenyl (C 22 ) are mentioned.

[0030] As used herein, the term "alkynyl," used alone or in compound words, refers to a straight-chain or branched-chain hydrocarbon residue containing at least one carbon-carbon triple bond. Preferably, the alkynyl group is a straight-chain alkynyl group. "C2-C 20 " is used to indicate the number of carbon atoms in the alkenyl group (in this case, 2 to 20).

[0031] As used herein, terms such as "heterocycle" or "heterocyclic group," used alone or in combination, refer to a saturated, partially unsaturated, or fully unsaturated monocyclic, bicyclic, or fused polycyclic ring system containing at least one heteroatom selected from the group consisting of nitrogen, sulfur, and oxygen. Prefixes such as "C5-C8" are used to indicate the number of carbon atoms (in this case, 5 to 8) in the cyclic portion of the group. Examples of suitable heterocyclic substituents include, but are not limited to, pyrrole, furan, benzofuran, benzothiazole, imidazole, benzimidazole, imidazoline, pyrazole, pyrazoline, triazole, oxazole, oxazoline, isoxazole, isoxazoline, furazan, oxadiazole, piperidine, pyridine, pyrimidine, pyridazine, and pyrazine, each of which may be further substituted with 1 to 3 substituents.

[0032] As used herein, terms such as "aryl" or "aromatic cyclic group" refer to any mononuclear or polynuclear linked or fused residue of an aromatic hydrocarbon ring system. Prefixes such as "C5-C8" are used to indicate the number of carbon atoms (in this case, 5 to 8) in the cyclic portion of the aryl group. Examples of aryl include phenyl (mononuclear), naphthyl (fused polynuclear), biphenyl (linked polynuclear), and tetrahydronaphthyl (fused polynuclear).

[0033] As used herein, the term "linker" refers to the moiety that bridges "X" to "L" and connects the drug to the glyceride unit in a compound of formula (I) as described herein.

[0034] As used herein, the term "self-immolative group" defines a chemical moiety that forms a severable bond with a linker and a stable bond with a drug, where the bond to the drug becomes unstable upon cleavage of the linker. Examples of self-immolative groups include, but are not limited to, acetal self-immolative groups, carboxyacetal self-immolative groups, carboxy(methyl acetal) self-immolative groups, para-hydroxybenzylcarbonyl self-immolative groups, flipped ester self-immolative groups, and trimethyllock self-immolative groups. Many other suitable groups are also suitable. Suitable self-immolative groups are known in the art, for example as described in CA Blencowe et al., Polym. Chem. 2011, 2, 773-790 and F. Kratz et al., ChemMedChem. 2008, 3(1), 20-53.

[0035] As used herein, the term "drug" refers to any pharmaceutically active agent or imaging agent (contrast agent) that would benefit from being transported through the intestinal lymphatic system, e.g., to avoid first-pass metabolism or for targeted delivery within the lymphatic system.

[0036] Examples of suitable pharmaceutically active agents include, but are not limited to, testosterone, mycophenolic acid (MPA), buprenorphine, estrogen (estrogens), opiates such as morphine, tetrahydrocannabinol (THC), cannabidiol, metoprolol, raloxifene, alphaxolone, statins such as atorvastatin, pentazocine, propranolol, L-DOPA, lidocaine, chlorpromazine, sertraline, amitriptyline, nortriptyline, pentazocine, glyceryl trinitrate, oxprenolol, laprolol, lanolin, lanolin, lanolin-100, lanolin-200, lanolin-300, lanolin-400, lanolin-500, lanolin-600, lanolin-700, lanolin-800, lanolin-900, lanolin-1000, lanolin-1100, lanolin-1200, lanolin-1300, lanolin-1400, lanolin-1500, lanolin-1600, lanolin-1700, lanolin-1800, lanolin-1900, lanolin-2000, lanolin-2100, lanolin-2200, lanolin-2300, lanolin-2400, lanolin-2500, lanolin-2600, lanolin-2700, lanolin-2800, lanolin-3000, lanolin-4000, lanolin-5000, lanolin-1900, lanolin-1900, lanolin-2900, lanolin-2900 Betalol, salbutamol, epitiostanol, melphalan, lovastatin, nonsteroidal anti-inflammatory drugs (NSAIDS, e.g., aspirin, ibuprofen, naproxen), COX-2 inhibitors (e.g., celecoxib), corticosteroid anti-inflammatory drugs (e.g., prednisolone, prednisone, dexamethasone), antimalarials (e.g., hydroxychloroquine), nitrosoureas, methotrexate, dactinomycin, anthracyclines (e.g., daunorubicin), mitomycin C, bleomycin, mithramycin, immunophilins Drugs that may be used for this purpose (e.g., cyclosporine, tacrolimus, sirolimus), sulfasalazine, leflunomide, mycophenolate, opioids, fingolimod, myriocin, chlorambucil, doxorubicin, nelarabine, cortisone, dexamethasone, prednisone, pralatrexate, vinblastine, bortezomib, nelarabine, daunorubicin hydrochloride, clofarabine, cytarabine, dasatinib, imatinib mesylate, ponatinib hydrochloride, vincristine sulfate, bendamustine hydrochloride, fludarabine phosphate, bosutinib, nilotinib, omasetake Simmepesuccinate, capecitabine, paclitaxel, gemcitabine, fulvestrant, tamoxifen, lapatinib, toremifene, ixabepilone, eribulin, albendazole, ivermectin, diethylcarbamazine, albendazole, doxycycline, closantel, maraviroc, enfuvirtide, deoxythymidine, zidovudine, stavudine, didanosine, zalcitabine, abacavir, lamivudine, emtricitabine, tenofovir, delavirdine, rilpivirine, raltegravir, elvitegravir, lopinavir, indinavir,These include nelfinavir, amprenavir, ritonavir, acyclovir, and pharmaceutically active peptides.

[0037] Examples of suitable imaging agents include, but are not limited to, fluorophores such as the Alexa Fluor series of optical imaging probes for fluorescence microscopy or for in vivo imaging where the fluorophore has an emission spectrum in the infrared range; gamma emitters that can be used for positron emission tomography (PET), such as fluorodeoxyglucose, or chelating agents for chelating magnetic resonance imaging probes, such as gadolinium or iron.

[0038] For the avoidance of any doubt, "Linear C 20 Reference to a "length equivalent to an alkyl group" refers to a theoretical length of 20 singly bonded carbon atoms.

[0039] In some preferred embodiments of the present invention, and with reference to general formula (I) or (II), one or more of the following definitions apply: a)R 1 and R 2 are independently H or C2-C 28 Represents a fatty acid residue. b)R 1 represents H, and R 2 is C2~C 28 Represents a fatty acid residue. c)R 2 represents H, and R 1 is C2~C 28 Represents a fatty acid residue. d)R 1 and R 2 and each represent palmitic acid. e) -X- is -O-. f) -X- is -NH-. g) -X- is -S-. h) -L- is -OC(O)-. i) -Y- is optionally substituted -C1 to C1- when -L- is -OC(O)-. 20 Alkyl C(O)OCH2-, -C2~C 20 Alkenyl C(O)OCH2- or -C2-C 20 represents an alkynyl C(O)OCH2- group, in which one or more of the carbon atoms of the alkyl, alkenyl, or alkynyl group may be replaced by NH, S, O, a C5-C8 aromatic or aliphatic cyclic group, or a C5-C8 aromatic or aliphatic heterocyclic group (provided that the alkyl, alkenyl, or alkynyl group is not a straight-chain C 20 provided that the length does not exceed that of the alkyl group. j) -Y- is -C1 to C1-C ... 20 Alkyl C(O)OCH2-, -C2~C 20 Alkenyl C(O)OCH2- or -C2-C 20 It represents an alkynyl C(O)OCH2- group. k) -Y- is -C1- ... 20 Alkyl C(O)OCH2-, -C2~C 20 Alkenyl C(O)OCH2- or -C2-C 20 It represents an alkynyl C(O)OCH2- group. l) -L- is -X'-. m) -Y- is an optionally substituted -C1-C2 alkylC(O)R 3 - group, or -C2 alkenyl C(O)R 3 - or -C2 alkynylC(O)R 3 - represents a group. n) -Y- is -C1-C2 alkylC(O)R optionally substituted with alkyl 3 - represents a group. o) -Y- is -C1-C2 alkylC(O)R optionally substituted by methyl 3 - represents a group. p)R 3is a self-immolative group selected from an acetal self-immolative group, a carboxyacetal self-immolative group, a carboxy(methylacetal) self-immolative group, a trimethyllock self-immolative group, a p-hydroxybenzylcarbonyl self-immolative group, or an inverted ester self-immolative group. n) X' is O. o) X' is S. p)X' is N(R 4 ) q) X' is N(H)S(O)2. r)R 4 is H. s)R 4 is C1-C4 alkyl. t)R 4 is methyl.

[0040] In one embodiment, L is X′ and —Y— is an optionally substituted —C1 alkylC(O)R 3 - represents.

[0041] Thus, in another embodiment, the present invention provides a compound of formula (I) represented by formula (II): [ka] [In the formula, R 1 , R 2 and -X- are as defined for formula (I), R 3 is a self-immolative group, [ka] represents the residue of a drug, -L- is -X'-; X' is O, S, N(R 4 ), or N(H)S(O)2, R 4 is H or C1-C4 alkyl, R 5 is selected from hydrogen and C1-C4 alkyl; or Pharmaceutically acceptable salts thereof are provided.

[0042] In another embodiment, L is X′ and —Y— is an optionally substituted —C2 alkylC(O)R 3 - represents.

[0043] Thus, in a further embodiment, the present invention provides a compound of formula (I) represented by formula (III): [ka] [In the formula, R 1 , R 2 and -X- are as defined for formula (I), R 3 is a self-immolative group, [ka] represents the residue of a drug, -L- is -X'-; X' is O, S, N(R 4 ), or N(H)S(O)2, R 4 is H or C1-C4 alkyl, R 5 and R 6 are independently selected from hydrogen and C1-C4 alkyl; or Pharmaceutically acceptable salts thereof are provided.

[0044] In another embodiment, the compound of formula (III) is selected from the compounds as listed in Table 1.

[0045] [Table 1]

[0046] In another embodiment, L is -OC(O)- and -Y- is an optionally substituted -C1-C 20 Alkyl C(O)OCH2-, -C2~C 20 Alkenyl C(O)OCH2- or -C2-C 20 represents an alkynyl C(O)OCH2- group, in which one or more of the carbon atoms of the alkyl, alkenyl, or alkynyl group may be replaced by NH, S, O, a C5-C8 aromatic or aliphatic cyclic group, or a C5-C8 aromatic or aliphatic heterocyclic group (provided that the alkyl, alkenyl, or alkynyl group is not a straight-chain C 20 provided that the length does not exceed that of the alkyl group.

[0047] Thus, in another embodiment, the present invention provides a compound of formula (I) represented by formula (IV): [ka] [In the formula, R 1 , R 2 and -X- are as defined for formula (I), [ka] represents the residue of a drug, R 5 and R 6 are independently selected from hydrogen and C1-C4 alkyl; R 7 is hydrogen or C1-C4 alkyl, n is 0 to 18; or Pharmaceutically acceptable salts thereof are provided.

[0048] In another embodiment, the compound of formula (IV) is selected from the compounds as listed in Table 2.

[0049] [Table 2]

[0050] In one embodiment, the agent is testosterone, or a derivative or analog thereof. Testosterone replacement therapy (TRT) is commonly used in patients with hypogonadism (a disorder characterized by abnormally low serum levels of testosterone) to restore the patient's serum testosterone levels to the normal range, thereby alleviating many of the symptoms of hypogonadism, such as mood disorders and sexual dysfunction.

[0051] Thus, in one embodiment, the present invention provides a compound of formula (I) represented by formula (V): [ka] [In the formula, R 1 , R 2 and -X- are as defined for formula (I), R 5 and R 6 are independently selected from hydrogen and C1-C4 alkyl; R 3 is a self-immolative group], or Pharmaceutically acceptable salts thereof are provided.

[0052] In another embodiment, the compound of formula (V) is selected from the compounds as listed in Table 3.

[0053] [Table 3]

[0054] In another embodiment, the present invention provides a method for treating or preventing a disease or disorder in which elevated testosterone levels are beneficial, comprising administering to a subject in need thereof a therapeutically effective The method includes administering an amount of a compound according to formula (V).

[0055] In a further embodiment, the present invention provides the use of a compound according to formula (V) in the manufacture of a medicament for treating or preventing a disease or disorder in which elevated testosterone levels are beneficial.

[0056] In yet another embodiment, the present invention provides a compound of formula (V) for use in the treatment or prevention of a disease or disorder in which elevated testosterone levels would be beneficial.

[0057] Diseases or disorders that may benefit from increased testosterone levels include, but are not limited to, hypogonadism, anemia due to bone marrow failure, anemia due to renal failure, chronic respiratory failure, chronic heart failure, steroid-dependent autoimmune disorders, AIDS wasting, hereditary angioedema or urticaria, end-stage breast cancer, or menopause.

[0058] In another embodiment, the present invention provides a method for enhancing lymphatic transport and systemic release of a drug, comprising: providing a pharmaceutical compound with a prodrug moiety of formula (VI): [ka] [In the formula, R 1 and R 2 are independently H or C2-C 28 represents a residue of a fatty acid, -X- is selected from -O-, -NH-, and -S-; -Y- is an optionally substituted -C1-C2 alkylC(O)R 3 - group, or -C2 alkenyl C(O)R 3 - or -C2 alkynylC(O)R 3 represents a - group, R 3 is a self-immolative group, [ka] indicates the point at which the linker is conjugated to the pharmaceutically active agent], or and conjugating a pharmaceutically acceptable salt thereof.

[0059] In one embodiment, with respect to the compounds as defined above, "Y" and "R 3 " will be selected to facilitate stable transport of the drug to the intestinal lymph. In another embodiment, Y and R 3 In yet another embodiment, Y and R will be selected to facilitate release of the drug in lymph, lymphocytes, lymphoid tissue, tissues with high lipase activity such as adipose tissue, certain cancers, the liver, or the systemic circulation. 3 is selected to facilitate stable transport of the drug to the intestinal lymph and release of the drug in lymph, lymphocytes, lymphoid tissue, tissues with high lipase activity such as adipose tissue, certain cancers, the liver, or systemic circulation.

[0060] The compounds of the present invention are useful for the stable transport of drugs to the intestinal lymph and the release of drugs in lymph, lymphocytes, lymphoid tissues, tissues with high lipase activity such as adipose tissue, certain cancers, the liver, or the systemic circulation. The compounds of the present invention are particularly useful for the transport and release of drugs for which avoidance of first-pass metabolism is beneficial, for example, compounds that exhibit a first-pass metabolic rate of greater than 50%. and release. In one embodiment, it is envisioned that the agent will exhibit a first-pass metabolic rate of greater than 60%. In another embodiment, the agent will exhibit a first-pass metabolic rate of greater than 70%. In a further embodiment, the agent will exhibit a first-pass metabolic rate of greater than 80%. In yet another embodiment, the agent will exhibit a first-pass metabolic rate of greater than 90%.

[0061] Drugs that may benefit from stable transport to the intestinal lymph and release in lymph, lymphocytes, lymphoid tissues, tissues with high lipase activity such as adipose tissue, certain cancers, the liver, or the systemic circulation include, but are not limited to, testosterone, mycophenolic acid, estrogen, morphine, tetrahydrocannabinol, cannabidiol, metoprolol, raloxifene, alphaxolone, statins such as atorvastatin, pentazocine, propranolol, L-DOPA, buprenorphine, midazolam, lidocaine, chlorpromazine, amitriptyline, nortriptyline, pentazocine, isosorbide dinitrate, glyceryl trinitrate, oxprenolol, labetalol, verapamil, salbutamol, epitiostanol, melphalan, lovastatin, and pharmaceutically active peptides.

[0062] The compounds of the invention are also useful for targeted release of drugs within the lymphatic system, e.g., lymph, lymphocyte, and lysogenic tissues, as well as tissues with high lipase activity such as adipose tissue, certain cancers, or the liver.

[0063] Drugs that may benefit from targeted release within the lymphatic system or in adipose tissue include, but are not limited to, nonsteroidal anti-inflammatory drugs (NSAIDS, e.g., aspirin, ibuprofen, naproxen), COX-2 inhibitors (e.g., celecoxib), corticosteroid anti-inflammatory drugs (e.g., prednisolone, dexamethasone), antimalarials (e.g., hydroxychloroquine), cyclophosphamide, PPAR agonists (e.g., fibrates), nitrosoureas, platinum, methotrexate, azathioprine, and the like. oprin, mercaptopurine, fluorouracil, dactinomycin, anthracyclines, mitomycin C, bleomycin, mithramycin, drugs acting on immunophilins (e.g., cyclosporine, tacrolimus, sirolimus), sulfasalazine, leflunomide, mycophenolate, opioids, fingolimod, myriocin, chlorambucil, doxorubicin, nelarabine, cortisone, dexamethasone, prednisone, pralatrexate, vinblastine, bortezomib, thiotepa, nerara vincristine sulfate, bendamustine hydrochloride, fludarabine phosphate, bosutinib, nilotinib, omacetaxine, mepesuccinate, anastrozole, capecitabine, letrozole, paclitaxel, gemcitabine, fulvestrant, tamoxifen, lapatinib, toremifene, ixabepilone, eribulin, albendazole, ivermectin, diethylcarbamazine, doxorubicin, daunorubicin hydrochloride, dasatinib, imatinib mesylate, ponatinib hydrochloride, vincristine sulfate, bendamustine hydrochloride, fludarabine phosphate, bosutinib, nilotinib, omacetaxine, mepesuccinate, anastrozole, capecitabine, letrozole, paclitaxel, gemcitabine, fulvestrant, tamoxifen, lapatinib, toremifene, ixabepilone, eribulin, albendazole, ivermectin, diethylcarbamazine, doxorubicin, These include cycycline, closantel, maraviroc, enfuvirtide, deoxythymidine, zidovudine, stavudine, didanosine, zalcitabine, abacavir, lamivudine, emtricitabine, tenofovir, nevirapine, delavirdine, efavirenz, rilpivirine, raltegravir, elvitegravir, lopinavir, indinavir, nelfinavir, amprenavir, ritonavir, acyclovir, and immunosuppressants such as mycophenolate, cyclosporine, tacrolimus, and sirolimus.

[0064] As a general strategy, the compounds of the present invention can be synthesized via one of the following routes.

[0065] [ka] Scheme 1. Synthesis of compounds of general formula (II).

[0066] Diacid chloride i, which is readily available from the corresponding malonic acid, can be reacted with diglyceride ii in the presence of pyridine to give acid-triglyceride (acid-TG) iii (see Scheme 1).

[0067] [ka] Scheme 2. Synthesis of compounds of general formula (III).

[0068] If acid anhydride iv is available, acid-TGiii can be generated by ring-opening with diglyceride ii in the presence of pyridine (Scheme 2). This method involves the R 5 and R 6 It works best when R 5 and R 6 are different from each other, a regioisomeric mixture of the acid-TGiii will result. Consequently, in this situation, other methods should be adopted, such as the method outlined in Scheme 3.

[0069] [ka] Scheme 3.R 5 =Me, R 6 Synthesis of compounds of formula (III) where =H.

[0070] R 5 =Me and R 6In the specific example where ═H, the known carboxylic acid v (Lienard, BM Ret al., Org. Biomol. Chem. 2008, 6, (13), 2282-2292) can be used as a starting point to obtain the acid-TGiii as a single regioisomer (see Scheme 3). Coupling of acid v with 1,3-DGii under standard conditions produces the TBDPS-protected triglyceride vi, which can be treated with TBAF and AcOH to afford the alcohol vii. A two-step oxidation process (PCC followed by KMnO) can then be used to convert the alcohol vii to the desired acid-TGiii via the intermediate aldehyde viii.

[0071] [ka] Scheme 4.R 3 Synthesis of compounds of general formula (III) where X'=O and X'=O is an acetal self-immolative (ASI) group.

[0072] When synthesizing compounds containing an acetal self-immolative (ASI) group between the drug and the alkyl spacer, the alcohol-bearing parent molecule must be functionalized and activated before conjugation with the acid-triglyceride iii, as outlined above in Scheme 4. Treatment of the alcohol with DMSO in a mixture of acetic anhydride and acetic acid forms the (methylthio)methyl (MTM) ether ix. Activation of the MTM ether ix with sulfuryl chloride forms the putative sulfoxide species, which can be reacted with the carboxylate of the acid-triglyceride iii to afford the target compound x.

[0073] [ka] Scheme 5.R 3is a carboxyacetal self-immolative (CASI) or carboxy(methyl acetal) self-immolative (CMSI) group, and X' is O or N(R 4 ) Synthesis of a compound of formula (III)

[0074] When a drug contains an alcohol, phenol, or amine (primary or secondary) functional group, a modified version of the acetal self-immolative group containing an additional carboxy group can be used. Reaction of the parent drug with a chloroalkyl chloroformate gives the chloroalkyl carbonate or carbamate xi (see Scheme 5). Displacement of the halogen leaving group is then achieved by treatment with the carboxylate derived from acid-TGiii in refluxing toluene to give the target compound xii.

[0075] [ka] Scheme 6.R 3 is a trimethyllock (TML) self-immolative group, and X'=O, N(R 4 ), or S(O)NH.

[0076] To synthesize prodrugs containing a trimethyl lock (TML) self-immolative group (Levine, MN; Raines, RTChem. Sci. 2012, 3, 2412-2420) between the drug and the alkyl spacer to facilitate systemic release of the parent molecule, acid-triglyceride iii must be functionalized with a TML moiety before conjugation with the drug, as outlined in Scheme 6. Coupling of acid-TGiii with TML phenol xiii under standard conditions affords triglyceride xiv, which can be deprotected under acidic conditions (10-camphorsulfonic acid) to afford alcohol xv. Sequential oxidation of alcohol xv to aldehyde xvi and then acid xvii can then be coupled to alcohol-, amine-, or sulfonamide-containing drugs under standard conditions to afford target compound xviii.

[0077] [ka] Scheme 7.R 3 is a p-hydroxybenzylcarbonyl (PHB) self-immolative group, and X'=O, S, or NR 4 Synthesis of a compound of formula (III)

[0078] To synthesize compounds containing a p-hydroxybenzyl (PHB) carbonyl self-immolative group, the primary hydroxyl group of p-hydroxybenzyl alcohol (xix) is first protected as a silyl ether, and the free phenolic hydroxyl group is coupled with the acid -TGiii to give PHB triglyceride xxi (see Scheme 7). After removal of the silicon protecting group, primary alcohol xxii can be activated by treatment with p-nitrophenyl (PNP) chloroformate to give PNP carbonate xxiii. Displacement of the PNP group is then achieved by reaction with an agent (A-X'H) under basic conditions to give the desired compound xxiv.

[0079] [ka] Scheme 8.R 3 is an inverted ester self-immolative (FSI) group, and X'=O, S, or NR 4 Synthesis of a compound of formula (III)

[0080] The inverted ester self-immolating (FSI) group is designed to liberate the free drug via a cyclization mechanism, resulting in the loss of a 4-carbon lactone (FSI-4) or 5-carbon lactone (FSI-5). FSI prodrugs can be synthesized by coupling the drug (A-X'H) with 4-bromobutyric acid (m=1) or 5-bromovaleric acid (m=2) (xxv) to give bromide xxvi (see Scheme 8). Displacement of bromide xxvi with the carboxylate derived from acid-TGiii generates the desired ester bond in target compound xxvii.

[0081] [ka] Scheme 9. Synthesis of compounds of formula (V).

[0082] If the agent contains a carboxylic acid, the acid -TGiii must be converted to the corresponding acid chloride, which is then reacted with trioxane (R 7 =H) or paraldehyde (R 7 =Me) affords the chloroalkyl ester xxviii (see Scheme 9). Displacement of the halide can then be achieved by reaction with a carboxylate derived from the parent drug to afford the target compound xxix.

[0083] If the compounds of the invention require purification, techniques such as recrystallization and chromatographic techniques, including high performance liquid chromatography (HPLC) and normal or reverse phase silica gel chromatography, can be used. Compounds can be characterized by nuclear magnetic resonance (NMR), mass spectrometry, and / or other suitable methods.

[0084] It will be understood that compounds of the present invention may exist in one or more stereoisomeric forms (e.g., diastereomers). The present invention includes within its scope all of these stereoisomeric forms, either isolated (e.g., enantiomerically isolated) or in combination (including racemic and diastereomeric mixtures).

[0085] Accordingly, the present invention also relates to substantially pure stereoisomeric forms of the compounds, e.g., in greater than about 90% diastereomeric excess, e.g., about 95%-97% diastereomeric excess, or greater than 99% diastereomeric excess, as well as mixtures, including racemic mixtures, thereof. Such diastereomers may be prepared, for example, by asymmetric synthesis using chiral intermediates, or mixtures may be resolved by conventional methods, e.g., chromatography or by use of a resolving agent.

[0086] When a compound contains one or more functional groups that may be protonated or deprotonated (for example, at physiological pH), the compound can be prepared and / or isolated as a pharmaceutically acceptable salt.It will be understood that the compound may be a zwitterion at a given pH.As used herein, the term "pharmaceutically acceptable salt" refers to a salt of a given compound that is suitable for pharmaceutical administration.Such salts can be formed by the reaction of an acid or a base with an amine group or a carboxylic acid group, respectively.

[0087] Pharmaceutically acceptable acid addition salts can be prepared from inorganic and organic acids. Examples of inorganic acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc. Examples of organic acids include acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, etc.

[0088] Pharmaceutically acceptable base addition salts can be prepared from inorganic and organic bases.The corresponding counterions derived from inorganic bases include sodium, potassium, lithium, ammonium, calcium, and magnesium salts.Organic bases include primary amines, secondary amines, and tertiary amines, substituted amines such as naturally occurring substituted amines, and cyclic amines such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-dimethylaminoethanol, tromethamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, N-alkylglucamine, theobromine, purine, piperazine, piperidine, and N-ethylpiperidine.

[0089] Acid / base addition salts tend to be more soluble in aqueous solvents than the corresponding free acid / base forms.

[0090] The compounds of the present invention may be in crystalline form or may be solvates (e.g., hydrates), and both forms are intended to be within the scope of the present invention. The term "solvate" refers to a complex of variable stoichiometry formed by a solute and a solvent. Suitable solvents should not interfere with the biological activity of the solute. Solvents may be, for example, water, ethanol, or acetic acid. Methods of solvation are generally known in the art.

[0091] The administration route of the compound of the present invention is intended to include oral administration and enteral administration.Therefore, the active compound can be formulated with an inert diluent or an absorbable edible carrier, or the active compound can be encapsulated in a hard or soft shell gelatin capsule, or the active compound can be compressed into a tablet, or the active compound can be directly incorporated into dietary food.For oral therapeutic administration, the active compound can be combined with an excipient and used in the form of ingestible tablets, buccal or sublingual tablets, lozenges, capsules, elixirs, suspensions, syrups, wafers, etc.The amount of the active compound in such therapeutically useful compositions is such that a suitable dosage is obtained.

[0092] Tablets, troches, pills, capsules, and the like may also contain the following components: binders, such as gum, acacia, corn starch, or gelatin; excipients, such as dicalcium phosphate; disintegrants, such as corn starch, potato starch, alginic acid, and the like; lubricants, such as magnesium stearate; and sweeteners, such as sucrose, lactose, or saccharin, or flavoring agents, such as peppermint, wintergreen oil, or cherry flavoring, may be added. When the unit dosage form is a capsule, it may contain a liquid carrier in addition to materials of the above type. Various other materials may be present as coatings or to otherwise modify the physical form of the dosage unit. For example, tablets, pills, or capsules may be coated with shellac, sugar, or both. A syrup or elixir may contain the active compound, sucrose as a sweetener, methylparaben and propylparaben as preservatives, a dye, and a flavoring such as cherry or orange flavor. Of course, any material used to prepare any unit dosage form should be pharmaceutically pure and substantially non-toxic in the amounts employed. In addition, the compounds of the present invention may be incorporated into sustained-release preparations and formulations, such as those that allow specific delivery of the drug to a specific region of the digestive tract. Liquid formulations can also be administered enterally, via the stomach or esophageal tube.

[0093] In one embodiment, the compounds of the invention will be administered orally with food to promote transport to the intestinal lymph.

[0094] In another embodiment, the compounds of the present invention will be co-administered orally with a lipid-based formulation to promote transport to the intestinal lymphatics, with or without the co-administration of food.

[0095] Lipid-based formulations for oral delivery are known in the art, and can include, for example, the substantially non-aqueous vehicle that typically contains one or more lipid components.Lipid vehicle and resulting lipid formulations can be usefully classified according to the common characteristics they share according to lipid formulation classification system (LFCS), as described below (Pouton, CW, Eur.J.Pharm.Sci.11(Supp 2),S93-S98,2000; Pouton, CW, Eur.J.Pharm.Sci.29,278-287,2006).

[0096] Thus, the lipid vehicle and resulting lipid formulation may contain oils / lipids and / or surfactants, optionally with a cosolvent. Type I formulations include mono-, di- and Type II formulations contain oils or lipids that require digestion, such as fatty acids and triglycerides, and combinations thereof. Type II formulations are water-insoluble self-emulsifying drug delivery systems (SEDDS) that contain the lipids and oils used in Type I formulations, along with an additional water-insoluble surfactant. Type III formulations are SEDDSs or self-microemulsifying drug delivery systems (SMEDDSs) that contain the lipids and oils used in Type I formulations, along with additional water-soluble surfactants and / or cosolvents (Type IIIa) or a larger proportion of water-soluble components (Type IIIb). Type IV formulations contain primarily hydrophilic surfactants and cosolvents (e.g., PEG, propylene glycol, and diethylene glycol monoethyl ether) and are useful for drugs that are poorly water-soluble but not lipophilic. Any such lipid formulations (Types I-IV) are contemplated in the present invention.

[0097] In some embodiments, the lipid vehicle contains one or more oils or lipids without additional surfactants, co-surfactants, co-emulsifiers, or co-solvents, i.e., essentially consists of one or more oils or lipids.In some further embodiments, the lipid vehicle contains one or more oils or lipids together with one or more water-insoluble surfactants, and optionally with one or more co-solvents.In some further embodiments, the lipid vehicle contains one or more oils or lipids together with one or more water-soluble surfactants, and optionally with one or more co-solvents.In some embodiments, the lipid vehicle contains a mixture of oil / lipid, surfactant, and co-solvent.In some embodiments, the lipid vehicle essentially consists of one or more surfactants / co-surfactants / co-emulsifiers, and / or solvents / co-solvents.

[0098] Examples of oils or lipids that can be used in the present invention include almond oil, babassu oil, blackcurrant seed oil, borage oil, canola oil, castor oil, coconut oil, cod liver oil, corn oil, cottonseed oil, evening primrose oil, fish oil, grape seed oil, mustard seed oil, olive oil, palm kernel oil, palm oil, peanut oil, rapeseed oil, safflower oil, sesame oil, shark liver oil, soybean oil, sunflower oil, walnut oil, wheat germ oil, avocado oil, rice bran oil, hydrogenated castor oil, hydrogenated coconut oil, hydrogenated cottonseed oil, hydrogenated palm oil, hydrogenated soybean oil, partially hydrogenated soybean oil, hydrogenated vegetable oil, caprylic / capric glycerides, fractionated triglycerides, triglycerides, sorbitan stearate, sorbitol ... Glyceryl caprate, glyceryl tricaproate, glyceryl tricaprylate, glyceryl tricaprylate / caprate, glyceryl tricaprylate / caprate, glyceryl tricaprylate / caprate / laurate, glyceryl tricaprylate / caprate / linoleate, glyceryl tricaprylate / caprate / stearate, glyceryl trilaurate, glyceryl monolaurate, glyceryl behenate, glyceryl monolinoleate, glyceryl trilinolenate, glyceryl trioleate, glyceryl triundecanoate, glyceryl tristearate linoleate, saturated polyglycolized glycerides, C8-C 12 Synthetic medium-chain triglycerides containing primarily fatty acid chains, C8-C 12 Medium chain triglycerides containing primarily fatty acid chains, >C 12 Examples of suitable triglycerides include long chain triglycerides containing primarily fatty acid chains of the formula (I), modified triglycerides, fractionated triglycerides, and mixtures thereof.

[0099] Examples of mono- and diglycerides that can be used in the present invention include glycerol monoesters and glycerol diesters having fatty acid chains of 8 to 40 carbon atoms, including hydrolyzed coconut oil (e.g., Capmul® MCM) and hydrolyzed corn oil (e.g., Maisine™ 35-1). In some embodiments, the mono- and diglycerides are mono- or di-saturated fatty acid esters of glycerol having fatty acid chains of 8 to 18 carbon atoms in length (e.g., glyceryl monostearate, glyceryl distearate, glyceryl monocaprylate, glyceryl dicaprylate, glyceryl monocaprate, and glyceryl dicaprate).

[0100] Suitable surfactants for use in lipid formulations include C8-C6 surfactants such as, but not limited to, propylene glycol monocaprylate, propylene glycol dicaprylate, and propylene glycol monolaurate, sold under trade names such as Capryol® 90, Labrafac® PG, and Lauroglycol® FCC. 22 Propylene glycol monoesters and diesters of fatty acids, including, but not limited to, sugar fatty acid esters such as sucrose palmitate, sucrose laurate, and sucrose stearate; sorbitan fatty acid esters such as, but not limited to, sorbitan laurate, sorbitan palmitate, and sorbitan oleate; polyoxyethylene sorbitan fatty acid esters such as, but not limited to, polysorbate 20, polysorbate 40, polysorbate 60, and polysorbate 80 and polysorbate 85; polyoxyethylene mono- and di-fatty acid esters such as, but not limited to, polyoxyl 40 stearate and polyoxyl 40 oleate; C8-C8 fatty acid esters, such as those sold under the trade names Labrasol®, Gelucire® 44 / 14, Gelucire® 50 / 13, and Labrafil®. 22 Polyoxyethylene monoesters and diesters of fatty acids and C8-C 22Mixtures of glyceryl monoesters, diesters, and triesters of fatty acids; polyoxyethylene castor oil compounds such as, but not limited to, Polyoxyl 35 castor oil, Polyoxyl 40 hydrogenated castor oil, and Polyoxyl 60 hydrogenated castor oil, as sold under the trade names Cremophor® / Kolliphor® EL, Cremophor® / Kolliphor® RH40, Cremophor® / Kolliphor® RH60, and the like; polyoxyethylene alkyl ethers such as, but not limited to, Polyoxyl 20 cetostearyl ether and Polyoxyl 10 oleyl ether; DL-alpha-tocopheryl polyethylene glycol succinate, as may be sold under the trade names; glyceryl monoesters, diesters, and triesters; C8-C 22 Glyceryl monoesters, diesters, and triesters of fatty acids; sucrose monoesters, diesters, and triesters; dioctyl sodium sulfosuccinate; polyoxyethylene-polyoxypropylene copolymers such as, but not limited to, Poloxamer 124, Poloxamer 188, and Poloxamer 407; C8-C10 alkyl acrylates such as, but not limited to, polyoxyethylene lauryl alcohol, polyoxyethylene cetyl alcohol, polyoxyethylene stearyl alcohol, and polyoxyethylene oleyl alcohol, sold under trade names such as Brij® 35, Brij® 58, Brij® 78, and Brij® 98. 22 and polyoxyethylene ethers of fatty alcohols, or a mixture of any two or more thereof.

[0101] Co-emulsifier or co-surfactant may be used in the formulation.Suitable co-emulsifier or co-surfactant may be phosphoglyceride; phospholipid such as lecithin, or free fatty acid that is liquid at room temperature, such as iso-stearic acid, oleic acid, linoleic acid, linolenic acid, palmitic acid, stearic acid, lauric acid, capric acid, caprylic acid, and caproic acid.

[0102] Suitable solvents / co-solvents include ethanol, propylene glycol, polyethylene glycol, diethylene glycol monoethyl ether, and glycerol.

[0103] Polymers can also be used in the formulation to prevent drug precipitation or to modify the drug release rate. A range of polymers have been shown to impart these properties and are well known to those skilled in the art. Suitable polymers include hydroxypropyl methylcellulose, hydroxypropyl methylcellulose acetylsuccinate, methyl other cellulose-derived polymers such as cellulose; poly(meth)acrylates such as the Eudragit series of polymers such as Eudragit E100, polyvinylpyrrolidone, or other polymers such as those described in, for example, Warren et al. Mol. Pharmaceuticals 2013, 10, 2823-2848.

[0104] Formulations can be selected to provide sustained release of the active substance in the gastrointestinal (GI) tract, particularly to control the rate of absorption. Many different approaches can be used to achieve these goals, such as the use of high-melting lipids that disperse / erode slowly in the GI tract, or polymers that form a slowly eroding matrix. These formulations may take the form of large monolithic dosage forms or may be present as micro- or nanoparticulate matrices, as described, for example, in Mishra, Handbook of Encapsulation and Controlled Release, CRC Press, Boca Raton, (2016) ISBN 978-1-4822-3234-9, Wilson and Crowley Controlled Release in Oral Drug Delivery, Springer, NY, ISBN 978-1-4614-1004-1 (2011), or Wise, Handbook of Pharmaceutical Controlled Release Technology, Marcel Dekker, NY, ISBN 0-82467-0369-3 (2000).

[0105] The formulation may also contain materials commonly known to those skilled in the art to be included in liquid-based formulations, such as antioxidants, e.g., butylated hydroxyanisole (BHA) or butylated hydroxytoluene (BHT), and solidifying agents such as microporous silica, e.g., magnesium aluminometasilicate (Neusilin).

[0106] In another embodiment, the compound may be orally co-administered with an enzyme inhibitor to increase the stability of the prodrug in the gastrointestinal tract or intestinal cells. In certain embodiments, the enzyme inhibitor is expected to inhibit pancreatic lipase, examples of which include, but are not limited to, Alli and Orlistat. In other embodiments, the enzyme inhibitor is expected to inhibit cellular lipase enzymes, such as monoacylglycerol lipase, examples of which include, but are not limited to, JZL184 (4-nitrophenyl-4-[bis(1,3-benzodioxol-5-yl)(hydroxy)methyl]piperidine-1-carboxylate).

[0107] While the compound or a pharmaceutically acceptable salt thereof as described above may be the only active ingredient administered to a subject, it is also within the scope of the present invention to administer other active ingredients together with this compound. In one or more embodiments, it is contemplated that a combination of two or more compounds of the present invention is administered to a subject.

[0108] The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of a compound as defined above, or a pharmaceutically acceptable salt thereof, together with at least one pharmaceutically acceptable carrier or diluent.

[0109] The term "composition" is also intended to include formulations in which the active ingredient is encapsulated in a carrier material, resulting in a capsule in which the active ingredient is surrounded by the carrier (with or without other carriers).

[0110] As one skilled in the art would readily understand, the nature of the pharmaceutically acceptable carrier will depend on the condition being treated and the nature of the mammal. The selection of a particular carrier or delivery system would be readily determined by one skilled in the art. Any formulation containing an active compound can be prepared using a suitable carrier. When preparing a compound, care should be taken to ensure that the activity of the compound is not destroyed in the process and that the compound can reach its site of action undestroyed. In some circumstances, it may be necessary to protect the compound by means known in the art, such as, for example, microencapsulation.

[0111] Those skilled in the art can easily determine the appropriate formulation for the compound of the present invention by using conventional approach.Identifying preferred pH range and suitable excipient, such as antioxidant, is common in the art.To provide desired pH range, buffer system is commonly used, including carboxylic acid buffer, such as acetate, citrate, lactate and succinate.A variety of antioxidants can be used for this formulation, including phenolic compounds such as BHT or vitamin E, reducing agents such as methionine or sulfite, and metal chelating agents such as EDTA.

[0112] Pharmaceutically acceptable vehicles and / or diluents include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic agents, and absorption delaying agents.The use of such media and agents for pharmaceutically active substances is well known in the art.Except where conventional media or agents are incompatible with active ingredients, their use in therapeutic compositions is contemplated.A supplementary active ingredient can also be incorporated into the composition.

[0113] The compound may also be administered in combination with one or more additional therapeutic agents. The combination may allow the compound as described above and other active ingredients to be administered separately, sequentially, or simultaneously. The combination may be provided in the form of a pharmaceutical composition.

[0114] The term "combination" as used herein refers to a composition or kit of parts, in which the combination partners as defined above may be administered dependently or independently, or by using different fixed combinations with different amounts of the combination partners, i.e., administered simultaneously or at different times.The combination partners may then be administered, for example, simultaneously or chronologically staggered, i.e., at different times, with equal or different time intervals for any part of the kit of parts.For example, the ratio of the total amounts of the combination partners administered in combination may vary to address the needs of a subpopulation of patients to be treated or the needs of a single patient.These different needs may be due to the age, sex, weight, etc. of the patient.

[0115] For ease of administration and uniformity of dosage, it is particularly advantageous to formulate compositions in unit dosage form.Unit dosage form, as used herein, refers to a physically separate unit that is suitable as a single dosage for the mammalian subject to be treated.Each unit contains a predetermined amount of active substance calculated to produce desired therapeutic effect, together with the necessary pharmaceutically acceptable vehicle.The specifications for the novel unit dosage form of the present invention are determined by and directly depend on (a) the unique characteristics of the active substance and the specific therapeutic effect to be achieved, and (b) the constraints inherent in the art of compounding active substance for the treatment of disease in living subjects with disease states that impair physical health as detailed herein disclosed.

[0116] As mentioned above, the main active ingredient can be formulated in a therapeutically effective amount in a unit dosage form with a suitable pharmaceutically acceptable vehicle for convenient and effective administration.The unit dosage form can contain the main active compound in an amount ranging from, for example, 0.25 μg to about 2000 mg.Expressed as a ratio, the active compound can be present in an amount of about 0.25 μg to about 2000 mg per mL of carrier.When the composition contains a supplementary active ingredient, the dosage is determined based on the usual dosage and the administration mode of the ingredient.

[0117] As used herein, the term "effective amount" refers to an amount of a compound that, when administered according to a desired dosing regimen, results in a desired therapeutic activity. Dosing may be administered once, at intervals of minutes or hours, or continuously over any one of these time periods. Suitable dosages may range from about 0.1 ng / kg body weight to 1 g / kg body weight per dose. Typical dosages are in the range of 1 μg / kg to 1 g / kg body weight per dose, e.g., 1 mg / kg to 1 g / kg body weight per dose. In one embodiment, dosages may range from 1 mg / kg to 500 mg / kg body weight per dose. In another embodiment, dosages may range from 1 mg / kg to 250 mg / kg body weight per dose. In yet another embodiment, dosages may range from 1 mg / kg to 100 mg / kg body weight per dose, e.g., up to 50 mg / kg body weight per dose.

[0118] The terms "treatment" and "treating," as used herein, encompass any treatment of a condition or disease in an animal, preferably a mammal, more preferably a human, including treatment of any disease or disorder in which elevated testosterone levels would be beneficial. The terms "prevention" and "preventing," as used herein, encompass the prevention or prophylaxis of a condition or disease in an animal, preferably a mammal, more preferably a human, including the prevention of any disease or disorder in which elevated testosterone levels would be beneficial.

[0119] Throughout this specification and the claims that follow, unless the context otherwise requires, the word "comprise" and variations such as "comprises" or "comprising" will be understood as implying the inclusion of a stated integer or group of integers or steps, but not the exclusion of any other integer or group of integers.

[0120] Reference herein to any prior publication (or information derived therefrom) or to any matter which is publicly known is not, and should not be construed as, an acknowledgment or understanding, or any form of suggestion, that the prior publication (or information derived therefrom) or publicly known matter forms part of the common general knowledge in the field to which this specification pertains. [Example]

[0121] The invention will now be described with reference to the following non-limiting examples, which are representative of general formula (I) and provide detailed methods for preparing exemplary compounds of the invention.

[0122] Example 1. Synthesis of acid-triglycerides from acid anhydrides. 4-((1,3-bis(palmitoyloxy)propan-2-yl)oxy)-4-oxobutanoic acid (iii) [ka]

[0123] To a solution of diglyceride ii (72.2 mg, 0.127 mmol) in pyridine (0.5 mL), CHCl (0.5 mL), and THF (0.5 mL) was added succinic anhydride (iv) (25.4 mg, 0.254 mmol) and DMAP (15.5 mg, 0.127 mmol), and the mixture was stirred at room temperature for 17 h. TLC analysis at this point indicated the presence of unreacted diglyceride, so additional succinic anhydride (25.4 mg, 0.254 mmol) and DMAP (15.5 mg, 0.127 mmol) were added, and the reaction was heated at 40° C. for an additional 22 h. The mixture was cooled to room temperature, diluted with ethyl acetate (20 mL), washed with 1 M HCl (10 mL) and brine (2×30 mL), dried (MgSO), and concentrated under reduced pressure to give the crude product. Purification by silica gel chromatography (15%-20%-25% ethyl acetate / hexanes) afforded acid-TGiii (77.0 mg, 91%) as a colorless solid.

[0124] 1 H NMR (400 MHz, CDCl3) δ 5.27 (m, 1H), 4.30 (dd, J = 12.0, 4.3 Hz, 2H), 4.15 (dd, J = 12.0, 5.8 Hz, 2H), 2.72 - 2.61 (m, 4H), 2.31 (t, J = 7.6Hz, 4H), 1.66 - 1.54 (m, 4H), 1.35 - 1.19 (m, 48H), 0.87 (t, J = 6.9 Hz, 6H).

[0125] 4-((1,3-bis(palmitoyloxy)propan-2-yl)oxy)-3 / 2-methyl-4-oxobutanoic acid (iii) [ka]

[0126] To a solution of diglyceride ii (200 mg, 0.351 mmol) and methylsuccinic anhydride (iv) (101 mg, 0.882 mmol) in pyridine / THF / CHCl (2.5 mL each) was added 4-(dimethylamino)pyridine (64.6 mg, 0.527 mmol), and the mixture was stirred at room temperature for 22 hours. The reaction was diluted with ethyl acetate (40 mL), washed with 1 M HCl (30 mL) and brine (3 × 30 mL), dried (MgSO), and concentrated under reduced pressure to give crude acid-TGiii (approximately 1:1 mixture of regioisomers, 240 mg, quantitative) as a colorless solid, which was used in the next reaction without purification.

[0127] 1 H NMR (400 MHz, CDCl3) δ 5.31 - 5.24 (m, 1H), 4.34 - 4.27 (m, 2H), 4.21 - 4.10 (m, 2H), 3.00 - 2.90 (m, 1H), 2.82 - 2.72 (m, 1H), 2.52 - 2.43 (m, 1H), 2.37 - 2.28 (m, 4H), 1.67 - 1.55 (m, 4H), 1.46 (d, J = 7.0 Hz, 3H), 1.36 - 1.17 (m, 48H), 0.88 (t, J = 6.8 Hz, 6H). Note: Integrals and multiplicities reflect the presence of a mixture of regioisomers.

[0128] Example 2.R 5 Synthesis of acid-triglycerides in which is a methyl group a) 2-((4-((tert-butyldiphenylsilyl)oxy)-2-methylbutanoyl)oxy)propane-1,3-diyldipalmitate (vi) [ka]

[0129] To a solution of acid v (175 mg, 0.491 mmol) and 1,3-DGii (293 mg, 0.515 mmol) in CHCl (8 mL) was added 4-(dimethylamino)pyridine (DMAP, 59.9 mg, 0.491 mmol) and EDC·HCl (235 mg, 1.23 mmol), and the mixture was stirred at room temperature for 16 h. The reaction was diluted with CHCl (20 mL), silica gel was added, and the mixture was concentrated under reduced pressure. Purification by silica gel chromatography (3%–10% ethyl acetate / hexanes) afforded triglyceride vi (406 mg, 91%) as a colorless oil.

[0130] 1 H NMR (400 MHz, CDCl3) δ 7.67 - 7.63 (m, 4H), 7.45 - 7.34 (m, 6H), 5.25 (m, 1H), 4.30 - 4.22 (m, 2H), 4.15 - 4.08 (m, 2H), 3.69 (t, J = 6.3 Hz, 2H), 2.75 (m, 1H), 2.27 (t, J = 7.2 Hz, 2H), 2.25 (t, J = 7.2 Hz, 2H), 1.99 (m, 1H), 1.64 - 1.55 (m, 5H), 1.32 - 1.20 (m, 48H), 1.14 (d, J = 7.2 Hz, 3H), 1.04 (s, 9H), 0.88 (t, J = 7.0 Hz, 6H).

[0131] b) 2-((4-hydroxy-2-methylbutanoyl)oxy)propane-1,3-diyldipalmitate (vii) [ka]

[0132] To a solution of TBDPS ether vi (406 mg, 0.447 μmol) in THF (20 mL) was added tetrabutylammonium fluoride (TBAF, 1.0 M in THF, 810 μL, 0.810 μmol) and acetic acid (46.1 mL, 0.810 mmol) at 0 °C, and the mixture was stirred at room temperature for 3 h. The reaction was diluted with water (30 mL) and extracted with ethyl acetate (3 × 20 mL). The organic extract was washed with brine (30 mL), dried (MgSO), and concentrated under reduced pressure to give the crude product. Purification by silica gel chromatography (0% to 25% ethyl acetate / hexanes) afforded alcohol vii (137 mg, 46%) as a colorless solid.

[0133] 1 H NMR (400 MHz, CDCl3) δ 5.27 (m, 1H), 4.39 (dd, J = 12.0, 4.1 Hz, 1H), 4.32 (dd, J = 11.9, 4.3 Hz, 1H), 4.16 (dd, J = 12.0, 6.0 Hz, 1H), 4.12 (dd, J = 12.0, 5.6 Hz, 1H), 3.74 - 3.63 (m, 2H), 2.66 (m, 1H), 2.316 (t, J = 7.6 Hz, 2H), 2.309 (t, J = 7.4 Hz, 2H), 1.91 (m, 1H), 1.70 (m, 1H), 1.63 - 1.58 (m, 4H), 1.34 - 1.25 (m, 48H), 1.19 (d, J = 7.1 Hz, 3H), 0.88 (t, J = 6.9Hz, 6H).

[0134] c) 2-((2-methyl-4-oxobutanoyl)oxy)propane-1,3-diyldipalmitate (viii) [ka]

[0135] To a suspension of alcohol vii (137 mg, 0.205 mmol) and Celite (90 mg) in CHCl (12 mL) was added pyridinium chlorochromate (PCC, 89.1 mg, 0.410 mmol) at 0 °C, and the mixture was stirred at room temperature for 2.5 h. The reaction was filtered through a short pad of silica gel, eluting with 50% ethyl acetate / hexanes, and the filtrate was concentrated under reduced pressure to give crude aldehyde viii (134 mg, quantitative) as a yellow oil, which was used without purification.

[0136] 1 H NMR (400 MHz, CDCl3) δ 9.75 (s, 1H), 5.27 (m, 1H), 4.30 (dd, J = 12.0, 4.2 Hz, 2H), 4.16 - 4.09 (m, 2H), 2.99 (m, 1H), 2.89 (ddd, J = 18.1, 7.8, 0.8 Hz, 1H), 2.55 (ddd, J = 18.0, 5.5, 0.9 Hz, 1H), 2.32 (t, J = 7.4 Hz, 2H), 2.30 (t, J = 7.4 Hz, 2H), 1.64 - 1.58 (m, 4H), 1.33 - 1.25 (m, 48H), 1.22 (d, J = 7.1 Hz, 3H), 0.88 (t, J = 7.0 Hz, 6H).

[0137] d) 4-((1,3-bis(palmitoyloxy)propan-2-yl)oxy)-3-methyl-4-oxobutanoic acid (iii) [ka]

[0138] To aldehyde viii (134 mg, 0.205 μmol) in acetone (7.5 mL) and water (2.5 mL) was added potassium permanganate (65.4 mg, 0.410 μmol), and the mixture was stirred at room temperature for 19 hours. The reaction was diluted with water (25 mL), acidified to pH 2 using 1 M HCl, and the aqueous layer was extracted with CHCl (3 × 20 mL). The combined organic extracts were washed with brine (30 mL), dried (MgSO), and concentrated under reduced pressure to give the crude product. Purification by silica gel chromatography (10% to 25% ethyl acetate / hexanes) gave acid iii (79.6 mg, 58%) as a colorless solid.

[0139] 1 H NMR (400 MHz, CDCl3) δ 5.27 (m, 1H), 4.32 - 4.27 (m, 2H), 4.18 - 4.12 (m, 2H), 2.92 (m, 1H), 2.78 (dd, J = 16.9, 8.0 Hz, 1H), 2.46 (dd, J = 16.9, 6.0 Hz, 1H), 2.304 (t, J = 7.6 Hz, 2H), 2.297 (t, J = 7.6 Hz, 2H), 1.62 - 1.56 (m, 4H), 1.31 - 1.19 (m, 51H), 0.88 (t, J = 6.8 Hz, 6H).

[0140] Example 3.R 3 Synthesis of compounds of general formula (III) where is an acetal self-immolative (ASI) group. 1,3-Bis(palmitoyloxy)propan-2-yl ((((8R,9S,10R,13S,14S,17S)-10,13-dimethyl-3-oxo-2,3,6,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclohexyl) Ropenta[a]phenanthren-17-yl)oxy)methyl)succinate (12) [ka]

[0141] To a solution of MTM ether ix (46.9 mg, 0.135 mmol) in CHCl (2 mL) was added a solution of sulfuryl chloride (13.3 μL, 0.164 mmol) in CHCl (1 mL) at 0 °C, and the mixture was stirred at 0 °C for 30 min, then at room temperature for an additional 1 h. The reaction was concentrated under a stream of N, coevaporated from toluene (2 × 5 mL), and dried under reduced pressure. The crude residue was then redissolved in toluene (1.5 mL) and added to a solution of acid iii (50.0 mg, 0.0747 mmol) and DBU (16.8 μL, 0.112 mmol) in toluene (1.5 mL) that had been previously stirred for 1 h, and the mixture was stirred at room temperature for 2 h. The reaction was diluted with CHCl (20 mL), and the organic phase was washed with saturated aqueous NaHCO (20 mL) and brine (20 mL), dried (MgSO), and concentrated under reduced pressure to give the crude product. Purification by silica gel chromatography (10%-25% ethyl acetate / hexanes with 0.5% EtN) gave compound 12 (8.4 mg, 12%) as a pale yellow oil.

[0142] 1 H NMR (400 MHz, CDCl3) δ 5.73 (s, 1H), 5.34 - 5.23 (m, 3H), 4.30 (dd, J = 11.9, 4.4 Hz, 2H), 4.15 (dd, J = 11.9, 5.8 Hz, 2H), 3.54 (dd, J = 8.3, 8.3 Hz, 1H), 2.65 (s, 4H), 2.48 - 2.24 (m, 8H), 2.09 - 1.99 (m, 2H), 1.92 - 1.80 (m, 2H), 1.74 - 1.36 (m, 9H), 1.35 - 1.21 (m, 49H), 1.19 (s, 3H), 1.17 - 0.91 (m, 5H), 0.88 (t, J = 6.9 Hz, 6H), 0.80 (s, 3H). ESI-HRMS: C 59 H 101 O 10 [M + H + ] calculated value 969.7389; measured value 969.7409.

[0143] 1-(1,3-bis(palmitoyloxy)propan-2-yl) 4-((((8R,9S,10R,13S,14S,17S)-10,13-dimethyl-3-oxo-2,3,6,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl)oxy)methyl) 2 / 3-methylsuccinate (13 / 14) [ka]

[0144] To a solution of MTM ether ix (76.3 mg, 0.219 mmol) in CHCl (3 mL) was added a solution of sulfuryl chloride (21.4 μL, 0.264 mmol) in CHCl (1.5 mL) at 0 °C and stirred at 0 °C for 30 min and then at room temperature for an additional 45 min. The reaction was concentrated under a stream of N2, coevaporated from toluene (2 × 5 mL), and dried under reduced pressure. The crude residue was then redissolved in toluene (2.5 mL) and added to a solution of acid iii (99.4 mg, 0.146 mmol) and DBU (32.8 μL, 0.220 mmol) in toluene (2.5 mL) that had been stirred for 45 min. The mixture was stirred at room temperature for 3 h. The reaction was diluted with ethyl acetate (20 mL), and the organic phase was washed with saturated aqueous NaHCO3 (2 × 20 mL) and brine (2 × 20 mL), dried (MgSO4), and concentrated under reduced pressure to give the crude product. Purification by silica gel chromatography (5%–15% ethyl acetate / hexanes with 0.5% Et3N) afforded compounds 13 and 14 (74.2 mg, 52%) as a pale yellow oil (1:1 mixture of regioisomers).

[0145] 1H NMR (400 MHz, CDCl3) δ 5.73 (s, 1H), 5.38 - 5.21 (m, 3H), 4.32 - 4.25 (m, 2H), 4.20 - 4.10 (m, 2H), 3.58 - 3.50 (m, 1H), 2.98 - 2.87 (m, 1H), 2.81 - 2.71 (m, 1H), 2.47 - 2.24 (m, 9H), 2.08 - 1.98 (m, 2H), 1.92 - 1.80 (m, 2H), 1.75 - 1.51 (m, 8H), 1.50 - 1.20 (m, 53H), 1.19 (s, 3H), 1.17 - 0.89 (m, 5H), 0.88 (t, J = 7.0 Hz, 6H), 0.80 (s, 1.5H), 0.79 (s, 1.5H). Note: The integrals and multiplicities are This reflects the presence of a 1:1 mixture of regioisomers in addition to the corresponding diastereoisomers.

[0146] 1-(1,3-bis(palmitoyloxy)propan-2-yl) 4-((((8R,9S,10R,13S,14S,17S)-10,13-dimethyl-3-oxo-2,3,6,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl)oxy)methyl) 2-methylsuccinate (13) [ka]

[0147] Prepared as above from the α-methyl regioisomer of acid-TGiii. 1 H NMR (400 MHz, CDCl3) δ 5.72 (s, 1H), 5.32 - 5.22 (m, 3H), 4.29 / 4.27 (each dd, J = 12.0, 4.2 Hz, 2H), 4.17 / 4.14 (each dd, J = 11.9, 6.0 Hz, 2H), 3.529 / 3.524 ( t, J = 8.3 Hz, 1H), 2.93 (m, 1H), 2.75 (dd, J = 16.8, 7.9 Hz, 1H), 2.49 - 2.23 (m, 9H), 2.08 - 1.96 (m, 2H), 1.91 - 1.79 (m, 2H), respectively. 1.71 (m, 1H), 1.67 - 1.51 (m, 8H), 1.49 - 1.07 (m, 51H), 1.22 (d, J = 7.2 Hz, 3H), 1.18 (s, 3H), 1.06 - 0.90 (m, 3H), 0.87 (t, J = 6.8 Hz, 6H), 0.79 (s, 3H). Note: Double signals (e.g., 4.29 / 4.27) reflects the presence of a mixture of diastereoisomers. ESI-HRMS: C 60 H 102 O 10 Na [M + Na + ] calculated value 1005.7365; measured value 1005.7370.

[0148] Example 4.R 3 Synthesis of compounds of general formula (III) where is a carboxy-acetal or carboxy-methylacetal self-immolative (CASI or CMSI) group. a) Chloromethyl ((1S,4S)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydronaphthalen-1-yl)(methyl)carbamate (xi) [ka]

[0149] To sertraline hydrochloride (50.0 mg, 0.146 mmol) in CHCl (4 mL) was added chloromethyl chloroformate (25.9 μL, 0.292 mmol) and pyridine (41.3 μL, 0.511 mmol) at 0 °C, and the mixture was stirred at 0 °C for 15 min and then at room temperature for 1 h. The reaction was diluted with CHCl (20 mL), and the organic phase was washed with saturated aqueous NaHCO (20 mL) and brine (20 mL), dried (MgSO), and concentrated under reduced pressure to give chloromethyl carbamate xi (58.2 mg, quantitative) as a pale yellow oil, which was used without purification.

[0150] 1 H NMR (400 MHz, CDCl3) δ 7.34 (d, J = 8.3 Hz, 1H), 7.31 - 7.25 (m, 1H), 7.24 - 7.16 (m, 1H), 7.11 - 7.06 (m, 1H), 6.97 (dd, J = 7.0, 2.0 Hz, 1H), 6.84 - 6.79 (m, 1H), 5.91 - 5.83 (m, 1H), 5.51 (dd, J = 10.4, 6.5 Hz, 0.6H), 5.32 (dd, J = 14.5, 6.1 Hz, 0.4H), 4.20 (dd, J = 5.2, 2.9 Hz, 1H), 2.78 (s, 1.2H), 2.72 (s, 1.8H), 2.36 - 2.22 (m, 1H), 2.07 - 1.99 (m, 1H), 1.87 - 1.70 (m, 2H). Note: The fractional integral reflects the presence of a mixture of rotamers.

[0151] b) 1,3-bis(palmitoyloxy)propan-2-yl(((((1S,4S)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydronaphthalen-1-yl)(methyl)carbamoyl)oxy)methyl)succinate (1) [ka]

[0152] To a suspension of acid-TGiii (25.1 mg, 37.5 μmol), chloromethyl carbamate xi (13.0 mg, 32.6 μmol), and tetrabutylammonium iodide (TBAI, 3.6 mg, 9.8 μmol) in toluene (1.2 mL) was added 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) (7.8 μL, 52.2 μmol), and the mixture was heated under reflux for 1 h. The reaction was cooled to room temperature, diluted with ethyl acetate (20 mL), and the organic phase was washed with water and brine (20 mL each), dried (MgSO), and concentrated under reduced pressure to give the crude product. Silica gel chromatography (15% ethyl acetate / hexanes) afforded compound 1 (29.0 mg, 86%) as a colorless oil.

[0153] 1 H NMR (400 MHz, CDCl3) δ 7.33 (d, J = 8.3 Hz, 1H), 7.30 - 7.24 (m, 1H), 7.22 - 7.17 (m, 2H), 7.09 (d, J = 2.0 Hz, 1H), 6.96 (d, J = 7.3 Hz, 1H), 6.84 - 6.78 (m, 1H), 5.89 - 5.83 (m, 2H), 5.49 (dd, J = 10.1, 6.6 Hz, 0.6H), 5.36 - 5.20 (m, 1.4H), 4.33 - 4.25 (m, 2H), 4.21 - 4.10 (m, 3H), 2.78 - 2.61 (m, 7H), 2.38 - 2.23 (m, 5H), 2.06 - 1.96 (m, 1H), 1.87 - 1.69 (m, 2H), 1.67 - 1.53 (m, 4H), 1.38 - 1.19 (m, 48H), 0.88 (t, J = 6.9 Hz, 6H). Note: The fractional integrals are based on the presence of a mixture of rotamers. It reflects the presence of ESI-HRMS: C 58 H 89 Cl2NO 10 Na [M + Na +] calculated value 1052.5756; measured value 1052.5774.

[0154] a2) 1-chloroethyl ((1S,4S)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydronaphthalen-1-yl)(methyl)carbamate (xi) [ka]

[0155] To sertraline hydrochloride (50.0 mg, 0.146 mmol) in CHCl (5 mL) was added 1-chloroethyl chloroformate (25.2 μL, 0.233 mmol) and pyridine (35.4 μL, 0.438 mmol) at 0 °C, and the mixture was stirred at 0 °C for 30 min and then at room temperature for 19 h. The reaction was diluted with CHCl (25 mL), and the organic phase was washed with saturated aqueous NaHCO (20 mL) and brine (20 mL), dried (MgSO), and concentrated under reduced pressure to give the crude product. Purification by silica gel chromatography (15% ethyl acetate / hexanes with 0.5% EtN) gave chloroethyl carbamate XI (50.4 mg, 84%) as a colorless solid.

[0156] 1 H NMR (400 MHz, CDCl3) δ 7.34 (d, J = 8.3 Hz, 1H), 7.31 - 7.24 (m, 1H), 7.24 - 7.16 (m, 2H), 7.13 - 7.04 (m, 1H), 7.00 - 6.94 (m, 1H), 6.85 - 6.77 (m, 1H), 6.75 - 6.64 (m, 1H), 5.54 - 5.46 (m, 0.6H), 5.41 - 5.33 (m, 0.4H), 4.20 (br s, 1H), 2.74 / 2.71 / 2.70 (respectively s, 3H), 2.36 - 2.24 (m, 1H), 2.09 - 1.97 (m, 1H), 1.90 - 1.71 (m, 5H). Note: Fractional integrals and doublets reflect the presence of both rotamers and diastereoisomers.

[0157] b2) 1,3-bis(palmitoyloxy)propan-2-yl(1-((((1S,4S)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydronaphthalen-1-yl)(methyl)carbamoyl)oxy)ethyl)succinate (2) [ka]

[0158] To a suspension of acid-TGiii (25.0 mg, 37.4 μmol), 1-chloroethyl carbamate xi (13.4 mg, 32.5 μmol), and tetrabutylammonium iodide (TBAI, 3.6 mg, 9.7 μmol) in toluene (1.2 mL) was added 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) (7.8 μL, 52.0 μmol). (μmol) was added and the mixture was heated under reflux for 1 h. The reaction was cooled to room temperature, diluted with ethyl acetate (30 mL), and the organic phase was washed with water and brine (20 mL each), dried (MgSO4), and concentrated under reduced pressure to give the crude product. Silica gel chromatography (4% to 8% ethyl acetate / toluene) gave compound 2 (19.7 mg, 58%) as a colorless oil.

[0159] 1H NMR (400 MHz, CDCl3) δ 7.33 (d, J = 8.3 Hz, 1H), 7.31 - 7.16 (m, 3H), 7.12 - 7.06 (m, 1H), 6.98 - 6.87 (m, 2H), 6.85 - 6.77 (m, 1H), 5.51 - 5.43 (m, 0.6H), 5.36 - 5.16 (m, 1.4H), 4.35 - 4.24 (m, 2H), 4.21 - 4.11 (m, 3H), 2.77 - 2.56 (m, 7H), 2.37 - 2.24 (m, 5H), 2.06 - 1.95 (m, 1H), 1.85 - 1.71 (m, 2H), 1.65 - 1.49 (m, 7H), 1.37 - 1.19 (m, 48H), 0.88 (t, J = 6.8 Hz, 6H).Note: Fractional integrals are This reflects the presence of both isomers and diastereoisomers. ESI-HRMS: C 59 H 91 Cl2NO 10 Na [M + Na + ] calculated value 1066.5912; measured value 1066.5957.

[0160] The following carboxy-acetal or carboxy-methyl acetal self-immolative containing compounds were prepared according to the above method: a3) Chloromethyl ((4aS,6R,7R,7aR,12bS)-3-(cyclopropylmethyl)-6-((S)-2-hydroxy-3,3-dimethylbutan-2-yl)-7-methoxy-1,2,3,4,5,6,7,7a-octahydro-4a,7-ethano-4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)carbonate (xi) [ka]

[0161] 1H NMR (400 MHz, CDCl3): δ 6.88 (d, J = 8.2 Hz, 1H), 6.61 (d, J = 8.2 Hz, 1H), 5.88 (s, 1H), 5.82 (d, J = 6.3 Hz, 1H), 5.72 (d, J = 6.3 Hz, 1H), 4.47 (d, J = 1.7 Hz, 1H), 3.48 (s, 3H), 3.06 - 2.98 (m, 2H), 2.89 (m, 1H), 2.63 (dd, J = 11.9, 5.0 Hz, 1H), 2.40 - 2.23 (m, 4H), 2.12 (t, J = 9.9 Hz, 1H), 2.03 - 1.79 (m, 3H), 1.71 (dd, J = 12.9, 2.5 Hz, 1H), 1.35 (s, 3H), 1.28 (m, 2H), 1.06 (m, 1H), 1.03 (s, 9H), 0.81 (m, 1H), 0.66 (m, 1H), 0.57 - 0.41 (m, 2H), 0.16 - 0.08 (m, 2H).

[0162] b3) 1,3-bis(palmitoyloxy)propan-2-yl((((((4aS,6R,7R,7aR,12bS)-3-(cyclopropylmethyl)-6-((S)-2-hydroxy-3,3-dimethylbutan-2-yl)-7-methoxy-1,2,3,4,5,6,7,7a-octahydro-4a,7-ethano-4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)carbonyl)oxy)methyl)succinate (4) [ka]

[0163] Compound 4 was prepared using an alternative method to that used above for compounds 1 and 2, as described below. Silver carbonate (3.1 mg, 11.2 μmol) was added to acid-TGiii (12.9 mg, 19.3 μmol) in DMF (0.6 mL), and the mixture was stirred at room temperature for 1 h. The reaction was concentrated under reduced pressure to give a gray residue. To this was added chloromethyl carbonate xi (9.0 mg, 16.1 μmol) in toluene (0.6 mL) and TBAI (1.8 mg, 4.8 μmol), and the mixture was heated under reflux for 1.5 h. The reaction was cooled to room temperature and then diluted with ethyl acetate (30 mL). The organic phase was washed with water (25 mL) and brine (25 mL), dried (MgSO), and concentrated under reduced pressure to give the crude product. Silica gel chromatography (5% to 15% ethyl acetate / hexanes) afforded compound 4 (3.7 mg, 19%) as a colorless solid.

[0164] 1 H NMR (400 MHz, CDCl3) δ 6.88 (d, J = 8.2 Hz, 1H), 6.60 (d, J = 8.1 Hz, 1H), 5.86 (s, 1H), 5.82 (s, 2H), 5.27 (m, 1H), 4.46 (s, 1H), 4.30 (dd, J = 12.0, 4.3 Hz, 2H), 4.15 (dd, J = 12.0, 5.9 Hz, 2H), 3.49 (s, 3H), 3.06 - 2.98 (m, 2H), 2.89 (m, 1H), 2.73 - 2.60 (m, 5H), 2.39 - 2.22 (m, 8H), 2.12 (t, J = 9.2 Hz, 1H), 2.01 (m, 1H), 1.93 - 1.80 (m, 2H), 1.75 - 1.49 (m, 5H), 1.35 (s, 3H), 1.34 - 1.20 (m, 49H), 1.05 (m, 1H), 1.03 (s, 9H), 0.88 (t, J = 6.9 Hz, 6H), 0.80 (m, 1H), 0.66 (m, 1H), 0.56 - 0.44 (m, 2H), 0.16 - 0.09 (m, 2H). ESI-HRMS: C 70 H 114 NO 14 [M + H + ] calculated value 1192.8234; measured value 1192.8244.

[0165] 1,3-Bis(palmitoyloxy)propan-2-yl(1-(((((4aS,6R,7R,7aR,12bS)-3-(cyclopropylmethyl)-6-((S)-2-hydroxy-3,3-dimethylbutan-2-yl)-7-methoxy-1,2,3,4,5,6,7,7a-octahydro-4a,7-ethano-4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)carbonyl)oxy)ethyl)succinate (5) [ka]

[0166] 1 H NMR (400 MHz, CDCl3) δ 6.88 / 6.87 (respectively d, J = 8.2 Hz, 1H), 6.77 (m, 1H), 6.59 (d, J = 8.2 Hz, 1H), 5.89 / 5.88 (respectively s, 1H), 5.25 (m, 1H), 4.46 (br s, 1H), 4.33 - 4.26 (m, 2H), 4.19 - 4.11 (m, 2H), 3.49 / 3.48 (s, 3H each), 3.06 - 2.98 (m, 2H), 2.89 (m, 1H), 2.74 - 2.57 (m, 5H), 2.38 - 2.20 (m, 8H), 2.12 (t, J = 10.0 Hz, 1H), 1.98 (td, J = 12.5, 5.5 Hz, 1H), 1.91 - 1.77 (m, 2H), 1.70 (dd, J = 13.4, 2.9 Hz, 1H), 1.64 - 1.52 (m, 7H), 1.35 (s, 3H), 1.33 - 1.18 (m, 49H), 1.06 (m, 1H), 1.03 (s, 5H), 0.88 (t, J = 6.9 Hz, 6H), 0.81 (m, 1H), 0.66 (m, 1H), 0.55 - 0.43 (m, 2H), 0.15 - 0.07 (m, 2H). Note: The double signal reflects the presence of a mixture of diastereoisomers. ESI-HRMS:C 71 H 116 NO 14 [M+H + ] calculated value 1206.8390, actual value 1206.8401.

[0167] 1,3-Bis(palmitoyloxy)propan-2-yl(1-(((((8R,9S,10R,13S,14S,17S)-10,13-dimethyl-3-oxo-2,3,6,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl)oxy)carbonyl)oxy)ethyl)succinate (20) [ka]

[0168] 1H NMR (400 MHz, CDCl3) δ 6.75 (m, 1H), 5.73 (s, 1H), 5.25 (m, 1H), 4.53 (m, 1H), 4.34 - 4.26 (m, 2H), 4.18 - 4.11 (m, 2H), 2.73 - 2.58 (m, 4H), 2.48 - 2.15 (m, 9H), 2.02 (m, 1H), 1.91 - 1.82 (m, 2H), 1.78 - 1.55 (m, 8H), 1.521 (d, J = 5.4 Hz, 1.5H), 1.517 (d, J = 5.4 Hz, 1.5H), 1.47 - 1.20 (m, 52H), 1.19 (s, 3H), 1.11 - 0.90 (m, 3H), 0.88 (t, J = 7.5 Hz, 6H), 0.86 (s, 3H). Note: The double signal is This reflects the presence of a mixture of diastereoisomers. ESI-HRMS: C 61 H 102 O 12 Na [M + Na + ] calculated value 1049.7263; measured value 1049.7273.

[0169] Example 5.R 3 Synthesis of compounds of general formula (III) where is a trimethyl lock (TML) self-immolative group. a) 1,3-bis(palmitoyloxy)propan-2-yl(2-(4-((tert-butyldimethylsilyl)oxy)-2-methylbutan-2-yl)-3,5-dimethylphenyl)succinate (xiv) [ka]

[0170] To a solution of acid-TGiii (100 mg, 0.149 mmol) and phenol xiii (53.0 mg, 0.164 mmol) in CHCl (4 mL) was added 4-(dimethylamino)pyridine (DMAP, 18.3 mg, 0.149 mmol) and EDC·HCl (71.6 mg, 0.374 mmol), and the mixture was stirred at room temperature for 19 h. The reaction was diluted with CHCl (10 mL), silica gel was added, and the mixture was concentrated under reduced pressure. Purification by silica gel chromatography (3%–7.5% ethyl acetate / hexanes) afforded TML triglyceride xiv (84.6 mg, 58%) as a colorless oil.

[0171] 1 H NMR (400 MHz, CDCl3) δ 6.80 (d, J = 2.0 Hz, 1H), 6.55 (d, J = 1.9 Hz, 1H), 5.29 (m, 1H), 4.31 (dd, J = 11.9, 4.4 Hz, 2H), 4.16 (dd, J = 12.0, 5.8 Hz, 2H), 3.51 - 3.44 (m, 2H), 2.85 (t, J = 6.9 Hz, 2H), 2.75 (t, J = 6.9 Hz, 2H), 2.51 (s, 3H), 2.30 (t, J = 7.6 Hz, 4H), 2.22 (s, 3H), 2.06 - 1.99 (m, 2H), 1.65 - 1.56 (m, 4H), 1.46 (s, 6H), 1.37 - 1.20 (m, 48H), 0.88 (t, J = 6.9 Hz, 6H), 0.84 (s, 9H), -0.03 (s, 6H).

[0172] b) 1,3-bis(palmitoyloxy)propan-2-yl (2-(4-hydroxy-2-methylbutan-2-yl)-3,5-dimethylphenyl) succinate (xv) [ka]

[0173] To the TBS ether xiv (83.7 mg, 86.0 μmol) in CHCl (1 mL) and MeOH (1 mL) was added 10-camphorsulfonic acid (3.0 mg, 12.9 μmol), and the mixture was stirred at room temperature for 1 h. The reaction was diluted with water (10 mL), and the aqueous layer was extracted with CHCl (3 × 10 mL). The combined organic extracts were washed with saturated aqueous NaHCO and brine (15 mL each), dried (MgSO), and concentrated under reduced pressure to give the crude product. Purification by silica gel chromatography (15%–25% ethyl acetate / hexanes) afforded alcohol xv (59.9 mg, 81%) as a colorless oil.

[0174] 1 H NMR (400 MHz, CDCl3) δ 6.81 (d, J = 2.0 Hz, 1H), 6.56 (d, J = 1.4 Hz, 1H), 5.28 (m, 1H), 4.30 (dd, J = 12.0, 4.4 Hz, 2H), 4.17 (dd, J = 12.0, 5.8Hz, 2H), 3 .51 (t, J = 6.8 Hz, 2H), 2.88 (t, J = 6.6 Hz, 2H), 2.75 (t, J = 6.6 Hz, 2H), 2.52 (s, 3H), 2.29 (t, J = 7.6 Hz, 4H), 2.22 (s, 3H), 2.05 (t, J = 7.4 Hz, 2H), 1.65 - 1.57 (m, 4H), 1.50 (s, 6H), 1.37 - 1.20 (m, 48H), 0.88 (t, J = 6.9 Hz, 6H).

[0175] c) 1,3-bis(palmitoyloxy)propan-2-yl(3,5-dimethyl-2-(2-methyl-4-oxobutan-2-yl)phenyl)succinate (xvi) [ka]

[0176] To a suspension of alcohol xv (59.9 mg, 0.0697 mmol) and Celite (30 mg) in CHCl (3 mL) was added pyridinium chlorochromate (PCC, 30.1 mg, 0.139 mmol) at 0 °C, and the mixture was stirred at room temperature for 2 h. The reaction was filtered through a short pad of silica gel, eluting with 50% ethyl acetate / hexane, and the filtrate was concentrated under reduced pressure to give crude aldehyde xvi (59.8 mg, quantitative) as a yellow oil, which was used without purification.

[0177] 1 H NMR (400 MHz, CDCl3) δ 9.54 (t, J = 2.6 Hz, 1H), 6.84 (d, J = 2.0 Hz, 1H), 6.60 (d, J = 1.4 Hz, 1H), 5.28 (m, 1H), 4.30 (dd, J = 12.0, 4.3 Hz, 2H), 4.16 (dd, J = 12.0, 5.8 Hz, 2H), 2.86 (t, J = 6.7 Hz, 2H), 2.83 (d, J = 2.6 Hz, 2H), 2.75 (t, J = 6.3 Hz, 2H), 2.53 (s, 3H), 2.30 (t, J = 7.6 Hz, 4H), 2.23 (s, 3H), 1.64 - 1.58 (m, 4H), 1.56 (s, 3H), 1.55 (s, 3H), 1.32 - 1.22 (m, 48H), 0.88 (t, J = 6.9 Hz, 6H).

[0178] d) 1,3-(2-((4-((1,3-bis(palmitoyloxy)propan-2-yl)oxy)-4-oxobutanoyl)oxy)-4,6-dimethylphenyl)-3-methylbutanoic acid (xvii) [ka]

[0179] To aldehyde xvi (59.8 mg, 69.7 μmol) in acetone (2.4 mL) and water (0.8 mL) was added potassium permanganate (12.2 mg, 76.7 μmol), and the mixture was stirred at room temperature for 17 h. The reaction was diluted with water (10 mL), acidified to pH 2 using 1 M HCl, and the aqueous layer was extracted with CHCl (3 × 15 mL). The combined organic extracts were washed with brine (30 mL), dried (MgSO), and concentrated under reduced pressure to give the crude product. Purification by silica gel chromatography (10%–25% ethyl acetate / hexanes) afforded acid xvii (30.4 mg, 50%) as a colorless solid.

[0180] 1 H NMR (400 MHz, CDCl3) δ 6.81 (d, J = 1.6 Hz, 1H), 6.58 (d, J = 1.4 Hz, 1H), 5.28 (m, 1H), 4.30 (dd, J = 11.9, 4.4 Hz, 2H), 4.16 (dd, J = 12.0, 5.8 Hz, 2H), 2.88 (t, J = 6.6 Hz, 2H), 2.84 (s, 2H), 2.75 (t, J = 6.6 Hz, 2H), 2.53 (s, 3H), 2.29 (t, J = 7.6 Hz, 4H), 2.22 (s, 3H), 1.64 - 1.58 (m, 4H), 1.57 (s, 6H), 1.34 - 1.20 (m, 48H), 0.88 (t, J = 6.8 Hz, 6H).

[0181] e) 1,3-bis(palmitoyloxy)propan-2-yl (2-(4-(((8R,9S,10R,13S,14S,17S)-10,13-dimethyl-3-oxo-2,3,6,7,8,9,10,11,12,13-14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl)oxy)-2-methyl-4-oxobutan-2-yl)-3,5-dimethylphenyl)succinate (15) [ka]

[0182] To a solution of acid xvii (29.0 mg, 33.2 μmol) in CHCl (1.2 mL), 4-(dimethylamino)pyridine (DMAP, 4.1 mg, 33.2 μmol), EDC·HCl (16.0 mg, 83.0 μmol), and testosterone (17.2 mg, 60.0 μmol) were added and the mixture was stirred at room temperature for 19 h. The reaction was diluted with CHCl (5 mL), silica gel was added, and the mixture was concentrated under reduced pressure. Purification by silica gel chromatography (12.5% to 20% ethyl acetate / hexanes) afforded compound 15 (15.0 mg, 40%) as a colorless solid.

[0183] 1 H NMR (400 MHz, CDCl3) δ 6.80 (d, J = 2.0 Hz, 1H), 6.57 (d, J = 1.9 Hz, 1H), 5.72 (s, 1H), 5.28 (m, 1H), 4.46 (dd, J = 9.1, 7.3 Hz, 1H), 4.31 (dd, J = 11.9, 4.4 Hz, 2H), 4.16 (dd, J = 11.9, 5.8 Hz, 2H), 2.88 (t, J = 6.7 Hz, 2H), 2.81 (d, J = 11.4 Hz, 2H), 2.76 (t, J = 6.9 Hz, 2H), 2.54 (s, 3H), 2.50 - 2.23 (m, 8H), 2.21 (s, 3H), 2.14 - 1.96 (m, 2H), 1.81 (m, 1H), 1.69 (m, 1H), 1.65 - 1.47 (m, 14H), 1.40 - 1.20 (m, 51H), 1.17 (s, 3H), 1.10 - 0.92 (m, 4H), 0.88 (t, J = 6.9 Hz, 6H), 0.66 (s, 3H). ESI-HRMS: C 71 H 115 O 11 [M + H +] calculated value 1143.8434; measured value 1143.8443.

[0184] The following trimethyl lock self-immolative group-containing compounds were prepared according to the above method: 1,3-Bis(palmitoyloxy)propan-2-yl(2-(4-(((1S,4S)-4-(3,4-dichlorophenyl)-1,2,3,4-tetra-hydronaphthalen-1-yl)(methyl)amino)-2-methyl-4-oxobutan-2-yl)-3,5-dimethylphenyl)succinate (3) [ka]

[0185] To a solution of sertraline hydrochloride (10.2 mg, 29.8 μmol) and acid xvii (20.0 mg, 22.9 μmol) in CHCl (1 mL), 4-(dimethylamino)pyridine (DMAP, 2.8 mg, 22.9 μmol), EDC·HCl (11.0 mg, 57.3 μmol), and triethylamine (8.0 μL, 57.3 μmol) were added and the mixture was stirred at room temperature for 16 h. The reaction was diluted with CHCl (5 mL), silica gel was added, and the mixture was concentrated under reduced pressure. Purification by silica gel chromatography (15%–25% ethyl acetate / hexanes) afforded compound 3 (22.2 mg, 83%) as a colorless solid.

[0186] 1H NMR (400 MHz, CDCl3) δ 7.32 (d, J = 8.3 Hz, 0.7H), 7.31 (d, J = 8.3 Hz, 0.3H), 7.25 - 7.11 (m, 2H), 7.08 - 7.02 (m, 1.3H), 6.97 - 6.85 (m, 1.7H), 6.83 - 6.79 (m, 1.7H), 6.73 (dd, J = 8.3, 2.0 Hz, 0.3H), 6.60 (d, J = 1.8 Hz, 0.7H), 6.57 (d, J = 1.7 Hz, 0.3H), 5.88 (dd, J = 10.5, 6.3 Hz, 0.7H), 5.24 (m, 1H), 4.96 (dd, J = 10.9, 5.7 Hz, 0.3H), 4.32 - 4.25 (m, 2H), 4.20 - 4.10 (m, 3H), 3.09 (d, J = 15.5 Hz, 0.7H), 3.01 (d, J = 8.2 Hz, 0.3H), 2.92 - 2.73 (m, 4.3H), 2.70 - 2.64 (m, 3H), 2.60 - 2.56 (m, 3.7H), 2.29 (t, J = 7.5 Hz, 4H), 2.23 (s, 2.1H), 2.21 (s, 0.9H), 2.21 (m, 1H), 1.95 (m, 1H), 1.69 (s, 2.1H), 1.66 (s, 0.9H), 1.62 (s, 3H), 1.70 - 1.52 (m, 6H), 1.34 - 1.20 (m, 48H), 0.88 (t, J = 6.9 Hz, 6H). Note: Minutes The existence of a mixture of heterosexual bodies is reflected in the numerical integration. ESI-HRMS: C 69 H 103 Cl2NO9Na [M + Na + ]The calculated value is 1182.6902; the measured value is 1182.6904.

[0187] 1,3-Bis(palmitoyloxy)propan-2-yl(2-(4-(((4aS,6R,7R,7aR,12bS)-3-(cyclopropyl-methyl)-6-((S)-2-hydroxy-3,3-dimethylbutan-2-yl)-7-methoxy-1,2,3,4,5,6,7,7a-octahydro-4a,7-ethano-4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)-2-methyl-4-oxobutan-2-yl)-3,5-dimethylphenyl)succinate (6) [ka]

[0188] 1 H NMR (400 MHz, CDCl3) δ 6.80 (d, J = 1.9 Hz, 1H), 6.61 - 6.57 (m, 2H), 6.53 (d, J = 8.1 Hz, 1H), 5.90 (s, 1H), 5.28 (m, 1H), 4.39 (s, 1H), 4.30 (dd, J = 11.9 , 4.4 Hz, 2H), 4.16 (dd, J = 11.9, 5.8 Hz, 2H), 3.37 (s, 3H), 3.06 (ABq, 2H), 3.01 - 2.81 (m, 5H), 2.76 (t, J = 6.7 Hz, 2H), 2.60 (dd, J = 11.6, 4.8 Hz, 1H), 2.55 (s, 3H), 2.29 (t, J = 7.6 Hz, 4H), 2.37 - 2.18 (m, 4H), 2.21 (s, 3H), 2.10 (t, J = 9.8 Hz, 1H), 1.95 (td, J = 12.6, 5.4 Hz, 1H), 1.89 - 1.74 (m, 2H), 1.71 - 1.51 (m, 11H), 1.33 (s, 3H), 1.45 - 1.14 (m, 49H), 1.02 (s, 9H), 0.88 (t, J = 6.9 Hz, 6H), 0.83 - 0.61 (m, 2H), 0.54 - 0.42 (m, 2H). 0.14 - 0.07 (m, 2H). ESI-HRMS: C 81 H 128 NO 13 [M + H + ] calculated value 1322.9380; measured value 1322.9404.

[0189] Example 6.R 3 Synthesis of compounds of general formula (III) where is a p-hydroxybenzyl (PHB) carbonyl self-immolative group. a) 1,3-bis(palmitoyloxy)propan-2-yl(4-(((tert-butyldimethylsilyl)oxy)methyl)-phenyl)succinate (xxi) [ka]

[0190] To a solution of acid-TGiii (200 mg, 0.300 mmol) and phenol xx (93.1 mg, 0.391 mmol) in CHCl (15 mL) was added 4-(dimethylamino)pyridine (DMAP, 48.0 mg, 0.393 mmol) and EDC·HCl (123 mg, 0.639 mmol), and the mixture was stirred at room temperature for 21 h. The reaction was diluted with CHCl (15 mL), silica gel was added, and the mixture was concentrated under reduced pressure. Purification by silica gel chromatography (5%–15% ethyl acetate / hexanes) afforded PHB triglyceride xxi (202 mg, 76%) as a colorless oil.

[0191] 1 H NMR (400 MHz, CDCl3) δ 7.34 - 7.29 (m, 2H), 7.07 - 7.02 (m, 2H), 5.30 (m, 1H), 4.72 (s, 2H), 4.31 (dd, J = 12.0, 4.3 Hz, 2H), 4.16 (dd, J = 12.0, 5.9 Hz, 2H), 2.88 (t, J = 6.9 Hz, 2H), 2.76 (t, J = 6.5 Hz, 2H), 2.29 (t, J = 7.6 Hz, 4H), 1.63 - 1.55 (m, 4H), 1.33 - 1.20 (m, 48H), 0.94 (s, 9H), 0.88 (t, J = 6.9 Hz, 6H), 0.09 (s, 6H).

[0192] b) 1,3-bis(palmitoyloxy)propan-2-yl(4-(hydroxymethyl)phenyl)succinate (xxii) [ka]

[0193] To the TBS ether xxi (181 mg, 0.203 mmol) in CHCl (2.5 mL) and MeOH (2.5 mL) was added 10-camphorsulfonic acid (8.0 mg, 0.0344 mmol), and the mixture was stirred at room temperature for 3.5 h. The reaction was diluted with CHCl (30 mL), and the organic phase was washed with saturated aqueous NaHCO (2 × 20 mL) and brine (20 mL), dried (MgSO), and concentrated under reduced pressure to give the crude product. Purification by silica gel chromatography (10% to 25% ethyl acetate / hexanes) afforded alcohol xxii (128 mg, 81%) as a colorless solid.

[0194] 1 H NMR (400 MHz, CDCl3) δ 7.40 - 7.35 (m, 2H), 7.11 - 7.06 (m, 2H), 5.30 (m, 1H), 4.69 (d, J = 5.9 Hz, 2H), 4.31 (dd, J = 12.0, 4.3 Hz, 2H), 4.16 (dd, J = 12.0, 5.9 Hz, 2H), 2.92 - 2.86 (m, 2H), 2.79 - 2.73 (m, 2H), 2.29 (t, J = 7.6 Hz, 4H), 1.64 - 1.55 (m, 4H), 1.34 - 1.19 (m, 48H), 0.88 (t, J = 6.9 Hz, 6H).

[0195] c) 1,3-bis(palmitoyloxy)propan-2-yl(4-((((4-nitrophenoxy)carbonyl)oxy)methyl)-phenyl)succinate (xxiii) [ka] To alcohol xxii (20.0 mg, 25.8 μmol) in CHCl (2 mL) was added 4-nitrophenyl chloroformate (8.4 mg, 41.6 μmol) and pyridine (3.8 μL, 47.0 μmol) at 0 °C, and the mixture was stirred at 0 °C for 30 min and then at room temperature for 4.5 h. The reaction was diluted with CHCl (30 mL), and the organic phase was washed with saturated aqueous NaHCO and brine (3 × 25 mL each), dried (MgSO), and concentrated under reduced pressure to give the crude product. Purification by silica gel chromatography (10%–20% ethyl acetate / hexanes) gave PNP carbonate xxiii (15.7 mg, 65%) as a colorless solid.

[0196] 1 H NMR (400 MHz, CDCl3) δ 8.31 - 8.25 (m, 2H), 7.48 - 7.43 (m, 2H), 7.41 - 7.36 (m, 2H), 7.17 - 7.11 (m, 2H), 5.29 (m, 1H), 5.28 (s, 2H), 4.32 (dd, J = 12.0, 4.3 Hz, 2H), 4.17 (dd, J = 12.0, 5.8 Hz, 2H), 2.93 - 2.87 (m, 2H), 2.79 - 2.73 (m, 2H), 2.30 (t, J = 7.6 Hz, 4H), 1.63 - 1.51 (m, 4H), 1.34 - 1.18 (m, 48H), 0.88 (t, J = 6.9 Hz, 6H).

[0197] d) 1,3-bis(palmitoyloxy)propan-2-yl(4-((((((8R,9S,10R,13S,14S,17S)-10,13-dimethyl-3-oxo-2,3,6,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl)oxy)carbonyl)oxy)methyl)phenyl)succinate (16) [ka]

[0198] Testosterone (48.0 mg, 0.166 mmol) in CH2Cl2 (10 mL) and To a solution of 13 and PNP carbonate xxiii (127 mg, 0.135 mmol), 4-(dimethylamino)pyridine (DMAP, 21.3 mg, 0.174 mmol) and DIPEA (7.1 μL, 0.0406 mmol) were added, and the mixture was stirred at room temperature for 5 days. The reaction was diluted with CHCl (20 mL), washed with 1 M HCl, water, and brine (20 mL each), dried (MgSO), and concentrated under reduced pressure to give the crude product. Purification by silica gel chromatography (5% ethyl acetate / toluene) gave compound 16 (20.4 mg, 14%) as a colorless solid.

[0199] 1 H NMR (400 MHz, CDCl3) δ 7.42 - 7.37 (m, 2H), 7.12 - 7.05 (m, 2H), 5.73 (s, 1H), 5.29 (m, 1H), 5.12 (s, 2H), 4.52 (t, J = 8.4 Hz, 1H), 4.31 (dd, J = 12.0, 4.3 Hz, 2H), 4.16 (dd, J = 12.0, 5.9 Hz, 2H), 2.89 (t, J = 6.7 Hz, 2H), 2.76 (t, J = 6.7 Hz, 2H), 2.47 - 2.34 (m, 3H), 2.29 (t, J = 7.6 Hz, 4H), 2.34 - 2.16 (m, 2H), 2.02 (m, 1H), 1.89 - 1.80 (m, 2H), 1.74 - 1.53 (m, 8H), 1.47 - 1.19 (m, 51H), 1.18 (s, 3H), 1.10 - 0.92 (m, 4H), 0.88 (t, J = 6.9 Hz, 6H), 0.85 (s, 3H). ESI-HRMS: C 66 H 105 O 12 [M + H + ] calculated value 1089.7601; measured value 1089.7617.

[0200] Example 7.R 3 Synthesis of compounds of general formula (III) where is an inverted ester self-immolative (FSI) group. a) (8R,9S,10R,13S,14S,17S)-10,13-dimethyl-3-oxo-2,3,6,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl 4-bromobutanoate (xxvi) [ka]

[0201] To a solution of testosterone (29.9 mg, 0.100 mmol) and 4-bromobutyric acid (xxv) (21.0 mg, 0.130 mmol) in CHCl (3 mL) was added 4-(dimethylamino)pyridine (DMAP, 15.5 mg, 0.130 mmol) and DCC (43.8 mg, 0.210 mmol), and the mixture was stirred at room temperature for 24 h. Another 0.6 equivalents of acid, 1 equivalent of DCC, and 0.6 equivalents of DMAP were added, and the mixture was stirred at room temperature for an additional 2 days. The reaction was diluted with CHCl (10 mL), silica gel was added, and the mixture was concentrated under reduced pressure. Purification by silica gel chromatography (25% ethyl acetate / hexanes) afforded bromide xxvi (26.7 mg, 59%) as a colorless solid.

[0202] 1 H NMR (400 MHz, CDCl3) δ 5.73 (s, 1H), 4.62 (dd, J = 9.1, 7.9 Hz, 1H), 3.47 (t, J = 6.5 Hz, 2H), 2.50 (td, J = 7.1, 1.0 Hz, 2H), 2.47 - 2.23 (m, 4H), 2.22 - 2.13 (m, 3H), 2.06 - 1.99 (m, 1H), 1.85 (m, 1H), 1.78 (m, 1H), 1.74 - 1.63 (m, 2H), 1.61 - 1.53 (m, 2H), 1.52 - 1.32 (m, 3H), 1.23 - 1.15 (m, 1H), 1.19 (s, 3H) 1.11 - 0.91 (m, 3H), 0.83 (s, 3H).

[0203] b) 1,3-bis(palmitoyloxy)propan-2-yl (4-(((8R,9S,10R,13S,14S,17S)-10,13-dimethyl-3-oxo-2,3,6,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1 H-Cyclopenta[a]phenanthren-17-yl)oxy)-4-oxobutyl)succinate (17) [ka]

[0204] To a suspension of acid-TGiii (30.9 mg, 46.2 μmol) and bromide xxvi (18.3 mg, 41.8 μmol) in toluene (1.5 mL), 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) (9.9 μL, 66.2 μmol) was added, and the mixture was heated under reflux for 21 h. The reaction was cooled to room temperature and then diluted with ethyl acetate (20 mL). The organic phase was washed with water (10 mL) and brine (10 mL), dried (MgSO), and concentrated under reduced pressure to give the crude product. Silica gel chromatography (15%-25% ethyl acetate / hexanes) afforded compound 17 (21.6 mg, 50%) as a colorless solid.

[0205] 1 H NMR (400 MHz, CDCl3) δ 5.73 (s, 1H), 5.26 (m, 1H), 4.62 (dd, J = 9.1, 7.9 Hz, 1H), 4.30 (dd, J = 11.9, 4.4 Hz, 2H), 4.15 (dd, J = 11.9, 5.8 Hz, 2H), 4.13 (t, J = 6.5 Hz, 2H), 2.69 - 2.58 (m, 4H), 2.47 - 2.25 (m, 9H), 2.19 (m, 1H), 2.07 - 1.91 (m, 3H), 1.85 (m, 1H), 1.78 (m, 1H), 1.75 - 1.45 (m, 9H), 1.44 - 1.21 (m, 59H), 1.19 (s, 3H), 1.16 - 0.91 (m, 5H), 0.88 (t, J = 6.9 Hz, 6H), 0.83 (s, 3H). ESI-HRMS: C 62 H 104 O 11 Na [M + Na + ] calculated value 1047.7471; measured value 1047.7460.

[0206] The following inverted ester self-immolative group-containing compounds were prepared according to the above method: 1,3-Bis(palmitoyloxy)propan-2-yl(5-(((8R,9S,10R,13S,14S,17S)-10,13-dimethyl-3-oxo-2,3,6,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl)oxy)-5-oxopentyl)succinate (18) [ka]

[0207] 1 H NMR (400 MHz, CDCl3) δ 5.73 (s, 1H), 5.26 (m, 1H), 4.61 (dd, J = 9.1, 7.9 Hz, 1H), 4.29 (dd, J = 11.9, 4.4 Hz, 2H), 4.15 (dd, J = 11.9, 5.8 Hz, 2H), 4.10 (t, J = 6.0 Hz, 2H), 2.68 - 2.58 (m, 4H), 2.47 - 2.25 (m, 10H), 2.18 (m, 1H), 2.02 (ddd, J = 13.3, 4.9, 3.3 Hz, 1H), 1.85 (m, 1H), 1.77 (m, 1H), 1.74 - 1.45 (m, 13H), 1.44 - 1.21 (m, 50H), 1.19 (s, 3H), 1.18 - 0.91 (m, 4H), 0.88 (t, J = 6.9H z, 6H), 0.83 (s, 3H). ESI-HRMS: C 63 H 107 O 11 Na [M + Na + ] calculated value 1061.7627; observed value 1061.7654.

[0208] 1-(1,3-bis(palmitoyloxy)propan-2-yl) 4-(5-(((8R,9S,10R,13S,14S,17S)-10,13-dimethyl-3-oxo-2,3,6,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl)oxy)-5-oxopentyl) 2-methylsuccinate (19) [ka]

[0209] 1 H NMR (400 MHz, CDCl3) δ 5.73 (s, 1H), 5.26 (m, 1H), 4.61 (dd, J = 9.1, 7.9 Hz, 1H), 4.33 - 4.24 (m, 2H), 4.21 - 4.04 (m, 4H), 2.91 (m, 1H), 2.76 / 2.72 (each dd, J = 13.9, 7.9 Hz, 1H), 2.47 - 2.26 (m, 11H), 2.18 (m, 1H), 2.02 (m, 1H), 1.84 (m, 1H), 1.77 (m, 1H), 1.74 - 1.44 (m, 13H), 1.43 - 1.14 (m, 51H), 1.223 / 1.214 (respectively d, J = 7.2 Hz, 3H), 1.19 (s, 3H), 1.11 - 0.92 (m, 3H), 0.88 (t, J = 6.9 Hz, 6H), 0.83 (s, 3H). NOTE: The double signal reflects the presence of a mixture of diastereoisomers. ESI-HRMS: C 64 H 109 O 11 [M + H + ] calculated value 1053.7964; observed value 1053.7984.

[0210] 1,3-Bis(palmitoyloxy)propan-2-yl(5-(((4aS,6R,7R,7aR,12bS)-3-(cyclopropylmethyl)-6-((S)-2-hydroxy-3,3-dimethylbutan-2-yl)-7-methoxy-1,2,3,4,5,6,7,7a-octahydro-4a,7-ethano-4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)-5-oxopentyl)succinate (7) [ka]

[0211] 1 H NMR (400 MHz, CDCl3) δ 6.77 (d, J = 8.1 Hz, 1H), 6.59 (d, J = 8.1 Hz, 1H), 5.88 (s, 1H), 5.26 (m, 1H), 4.42 (d, J = 1.6 Hz, 1H), 4.29 (dd, J = 11.9, 4.4Hz, 2H), 4.15 (dd, J = 12.0, 5.8 Hz, 2H), 4.11 (t, J = 6.0 Hz, 2H), 3.45 (s, 3H), 3.02 (d, J = 13.5 Hz, 1H), 2.99 (s, 1H), 2.88 (m, 1H), 2.67 - 2.54 (m, 7H), 2.38 - 2.22 (m, 8H), 2.11 (t, J = 10.1 Hz, 1H), 1.97 (td, J = 12.7, 5.6 Hz, 1H), 1.91 - 1.84 (m, 2H), 1.82 - 1.68 (m, 5H), 1.65 - 1.57 (m, 4H), 1.35 (s, 3H), 1.35 - 1.19 (m, 49H), 1.05 (m, 1H), 1.03 (s, 9H), 0.88 (t, J = 6.9 Hz, 6H), 0.84 - 0.76 (m, 1H), 0.75 - 0.63 (m, 1H), 0.55 - 0.43 (m, 2H), 0.17 - 0.08 (m, 2H). ESI-HRMS: C 73 H 120 NO 13 [M + H + ] calculated value 1218.8754; measured value 1218.8775.

[0212] 1-(1,3-bis(palmitoyloxy)propan-2-yl) 4-(5-(((4aS,6R,7R,7aR,12bS)-3-(cyclopropylmethyl)-6-((S)-2-hydroxy-3,3-dimethylbutan-2-yl)-7-methoxy-1,2,3,4,5,6,7,7a-octahydro-4a,7-ethano-4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)-5-oxopentyl) 2-methylsuccinate (8) [ka]

[0213] 1 H NMR (400 MHz, CDCl3) δ 6.77 (d, J = 8.1 Hz, 1H), 6.59 (d, J = 8.1 Hz, 1H), 5.88 (s, 1H), 5.26 (m, 1H), 4.42 (d, J = 1.5 Hz, 1H), 4.32 - 4.24 (m, 2H), 4.20 - 4.06 (m, 4H), 3.45 (s, 3H), 3.02 (d, J = 13.3 Hz, 1H), 2.99 (s, 1H), 2.96 - 2.84 (m, 2H), 2.74 (ddd, J = 16.6, 13.4, 8.0 Hz, 1H), 2.66 - 2.53 (m, 3H), 2.46 - 2.21 (m, 9H), 2.11 (t, J = 9.9 Hz, 1H), 1.97 (td, J = 12.5, 5.6 Hz, 1H), 1.92 - 1.67 (m, 7H), 1.65 - 1.56 (m, 4H), 1.35 (s, 3H), 1.34 - 1.17 (m, 49H), 1.222 (d, J = 7.2 Hz, 1.5H), 1.116 (d, J = 7.2 Hz, 1.5H), 1.05 (m, 1H), 1.03 (s, 9H), 0.88 (t, J = 6.8 Hz, 6H), 0.80 (m, 1H), 0.67 (m, 1H), 0.55 - 0.42 (m, 2H), 0.15 - 0.08 (m, 2H). ESI-HRMS: C 74 H 122 NO 13 [M + H + ] calculated value 1232.8911; measured value 1232.8925.

[0214] 1,3-Bis(palmitoyloxy)propan-2-yl(5-((1-(isopropylamino)-3-(4-(2-methoxyethyl)-phenoxy)propan-2-yl)oxy)-5-oxopentyl)succinate (9) [ka]

[0215] To the Boc-protected prodrug xxvii (9.2 mg, 8.2 μmol) in CHCl (1.2 mL) was added trifluoroacetic acid (TFA, 6.1 μL, 82.2 μmol) at 0°C, and the mixture was stirred at room temperature for 21 h. The reaction was concentrated under a stream of N gas to give the crude product. Silica gel chromatography (1% to 3.5% methanol / CHCl) afforded the crude product. 2Cl2) to give compound 9 (6.8 mg, 81%) as a pale yellow oil.

[0216] 1 H NMR (400 MHz, CDCl3) δ 7.13 (d, J = 8.6 Hz, 2H), 6.84 - 6.77 (m, 2H), 5.38 (m, 1H), 5.24 (m, 1H), 4.28 (dd, J = 11.9, 4.5 Hz, 2H), 4.18 - 4.10 (m, 4H), 4.06 (t, J = 5.7 Hz, 2H), 3.55 (t, J = 7.0 Hz, 2H), 3.34 (s, 3H), 3.38 - 3.24 (m, 3H), 2.81 (t, J = 7.0 Hz, 2H), 2.66 - 2.56 (m, 4H), 2.48 - 2.35 (m, 2H), 2.31 (t, J = 7.6 Hz, 4H), 1.69 - 1.54 (m, 8H), 1.32 (d, J = 6.4 Hz, 6H), 1.37 - 1.19 (m, 48H), 0.88 (t, J = 6.9 Hz, 6H). ESI-HRMS: C 59 H 104 NO 12 [M + H + ] calculated value 1018.7553; measured value 1018.7568.

[0217] Example 8. Synthesis of a compound of general formula (IV) where the drug is mycophenolic acid (MPA). a) 1-(1,3-bis(palmitoyloxy)propan-2-yl) 5-(chloromethyl) 3-methylpentanedioate (xviii) [ka]

[0218] A mixture of acid-TGiii (75.0 mg, 0.108 mmol), N,N-dimethylformamide (DMF, 1 drop), and SOCl (78.0 μL, 1.08 mmol) was heated under reflux for 45 min and then cooled to room temperature. The reaction was concentrated under reduced pressure and then coevaporated three times from toluene (3 mL each) and dried under reduced pressure. The resulting acid chloride was redissolved in CHCl (1 mL) and added dropwise to anhydrous ZrCl (25.1 mg, 0.108 mmol) in CHCl (0.5 mL). The mixture was stirred at room temperature for 15 min and then cooled to 0 °C. 1,3,5-Trioxane (9.7 mg, 0.108 mmol) was added, and the mixture was stirred at room temperature for 20 h. The reaction was diluted with CHCl (15 mL), and the organic phase was washed with water and brine (15 mL each), dried (MgSO), and concentrated under reduced pressure to give the crude product. Purification by silica gel chromatography (5% to 12.5% ethyl acetate / hexanes) gave the chloromethyl ester xxviii (19.2 mg, 24%) as a yellow oil.

[0219] 1 H NMR (400 MHz, CDCl3) δ 5.72 - 5.67 (m, 2H), 5.27 (m, 1H), 4.31 (dd, J = 11.9, 4.2 Hz, 2H), 4.13 (dd, J = 12.0, 6.1 Hz, 2H), 2.53 - 2.34 (m, 5H), 2.31 (t, J = 7.6 Hz, 4H), 1.67 - 1.53 (m, 4H), 1.37 - 1.19 (m, 48H), 1.05 (d, J = 6.5 Hz, 3H), 0.88 (t, J = 6.9 Hz, 6H).

[0220] b) (E)-1-(((6-(4-(allyloxy)-6-methoxy-7-methyl-3-oxo-1,3-dihydroisobenzofuran-5-yl)-4-methylhex-4-enoyl)oxy)methyl) 5-(1,3-bis(palmitoyloxy)propan-2-yl) 3-methylpentanedioate (xxxi) [ka]

[0221] To a suspension of MPA(OAll)xxx (12.9 mg, 35.7 μmol), chloromethyl ester xxviii (19.0 mg, 25.5 μmol), and TBAI (4.7 mg, 12.7 μmol) in toluene (0.8 mL) was added 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) (6.9 μL, 45.9 μmol), and the mixture was heated at 80 °C for 2 h. The reaction was cooled to room temperature and then diluted with ethyl acetate (20 mL). The organic phase was washed with water and brine (20 mL each), dried (MgSO), and concentrated under reduced pressure to give the crude product. Silica gel chromatography (15%-20% ethyl acetate / hexanes) afforded the protected prodrug xxxi (16.7 mg, 61%) as a colorless solid.

[0222] 1 H NMR (400 MHz, CDCl3) δ 6.09 (m, 1H), 5.69 (s, 2H), 5.36 (m, 1H), 5.31 - 5.22 (m, 2H), 5.18 (td, J = 6.7, 1.2 Hz, 1H), 5.13 (s, 2H), 4.78 (dt, J = 5.9, 1.2 Hz, 2H), 4.292 / 4.288 (dd, J = 11.9, 4.3 Hz, 2H, respectively), 4.13 (dd, J = 11.9, 5.9 Hz, 2H), 3.76 (s, 3H), 3.41 (d, J = 6.7 Hz, 2H), 2.50 - 2.37 (m, 5H), 2.34 - 2.23 (m, 8H), 2.18 (s, 3H), 1.77 (s, 3H), 1.64 - 1.56 (m, 4H), 1.35 - 1.18 (m, 48H), 1.02 (d, J = 6.4 Hz, 3H), 0.87 (t, J = 6.9 Hz, 6H). Note: The double signal is due to the diaste This reflects the presence of a mixture of rheoisomers.

[0223] c) (E)-1-(1,3-bis(palmitoyloxy)propan-2-yl) 5-(((6-(4-hydroxy-6-methoxy-7-methyl-3-oxo-1,3-dihydroisobenzofuran-5-yl)-4-methylhex-4-enoyl)oxy)methyl) 3-methyl-pentanedioate (10) [ka]

[0224] To the allyl ether xxxi (16.7 mg, 15.6 μmol) in CHCl (0.5 mL) was added 1,3-dimethylbarbituric acid (4.9 mg, 31.2 μmol) and Pd(PPh) (5.4 mg, 4.7 μmol), and the mixture was stirred at room temperature for 2 h. The reaction mixture was applied directly to a short pad of silica gel and eluted with 50% ethyl acetate / hexane. The eluent was concentrated under reduced pressure to give the crude product, which was purified by silica gel chromatography (5%-10% ethyl acetate / toluene) to give compound 10 (10.1 mg, 63%) as a colorless solid.

[0225] 1 H NMR (400 MHz, CDCl3) δ 7.68 (s, 1H), 5.70 (s, 2H), 5.30 - 5.21 (m, 2H), 5.20 (s, 2H), 4.296 / 4.291 (dd, J = 11.9, 4.3 Hz, 2H, respectively), 4.13 (dd, J = 11.9, 6.0 Hz, 2H), 3.76 (s, 3H), 3.38 (d, J = 6.9 Hz, 2H), 2.49 - 2.38 (m, 5H), 2.34 - 2.25 (m, 8H), 2.15 (s, 3H), 1.79 (s, 3H), 1.66 - 1.54 (m, 4H), 1.35 - 1.19 (m, 48H), 1.02 (d, J = 6.4 Hz, 3H), 0.88 (t, J = 6.9 Hz, 6H). Note: Double signals indicate This reflects the presence of a mixture of stereoisomers. ESI-HRMS: C 59 H 97 O 14 [M + H + ] calculated value 1029.6873; measured value 1029.6890.

[0226] The following compounds of general formula (IV) were prepared according to the above method: a2) 1-(1,3-bis(palmitoyloxy)propan-2-yl) 5-(1-chloroethyl) 3-methylpentanedioate (xxviii) [ka]

[0227] A mixture of acid-TGiii (120 mg, 0.172 mmol), N,N-dimethylformamide (DMF, 1 drop), and SOCl (125 μL, 1.72 mmol) was heated under reflux for 1.25 h and then cooled to room temperature. The reaction was concentrated under reduced pressure and then coevaporated three times from toluene (3 mL each) and dried under reduced pressure. The resulting acid chloride was redissolved in CHCl (1.5 mL) and added dropwise to anhydrous ZnCl (23.5 mg, 0.172 mmol) in CHCl (0.5 mL) at 0 °C and stirred for 5 min at 0 °C. Paraldehyde (45.6 μL, 0.344 mmol) was added, and the mixture was stirred at 0 °C for 10 min and at room temperature for 1 h. The reaction was diluted with CHCl (20 mL), and the organic phase was washed with water and brine (20 mL each), dried (MgSO), and concentrated under reduced pressure to give the crude product. Purification by silica gel chromatography (5% to 15% ethyl acetate / hexanes) gave 1-chloroethyl ester xxviii (14.9 mg, 11%) as a yellow oil.

[0228] b2) (E)-1-(1,3-bis(palmitoyloxy)propan-2-yl) 5-(1-((6-(4-hydroxy-6-methoxy-7-methyl-3-oxo-1,3-dihydroisobenzofuran-5-yl)-4-methylhex-4-enoyl)oxy)ethyl) 3-methylpentanedioate (11) [ka]

[0229] 1H NMR (401 MHz, CDCl3) δ 7.68 (s, 1H), 6.80 (q, J = 5.4 Hz, 1H), 5.29 - 5.21 (m, 2H), 5.20 (s, 2H), 4.32 - 4.26 (m, 2H), 4.132 / 4.127 (respectively dd, J = 11.9, 6.0 Hz, 2H), 3.76 (s, 3H), 3.38 (d, J = 6.9 Hz, 2H), 2.48 - 2.35 (m, 5H), 2.32 - 2.18 (m, 8H), 2.15 (s, 3H), 1.79 (s, 3H), 1.64 - 1.54 (m, 4H), 1.41 (d, J = 5.4 Hz, 3H), 1.36 - 1.18 (m, 48H), 1.01 (d, J = 6.4 Hz, 3H), 0.88 (t, J = 6.9 Hz, 6H).Note : The double signal reflects the presence of a mixture of diastereoisomers. ESI-HRMS: C 60 H 99 O 14 Na [M + Na + ] calculated value 1065.6849; measured value 1065.6879.

[0230] Example 9. Lymphatic transport studies in rats In order to evaluate the lymphatic transport of the compound of the present invention in rats, cannula is inserted into the mesenteric lymphatic vessel of rats, so that mesenteric lymph can be continuously collected.Then, the lipid formulation containing the compound of interest is administered to animals.After collecting lymph, the drug concentration in lymph is quantified.

[0231] Lipid-based formulations of the compounds of the present invention or control compounds were prepared as previously described (Trevaskis, NLet et al., Pharmaceutical Research, 2005, 22(11), 1863-1870). Briefly, approximately 2 mg of compound (1.5 mg for compound 19 and 0.05 mg for compounds 6 and 7), 40 mg of oleic acid, and 25 mg of Tween 80 were mixed in a glass vial until equilibrated (gentle heating (less than 50 ° C) may be applied for a short time). Subsequently, an aqueous phase consisting of 5.6 mL of phosphate-buffered saline (PBS, pH 7.4) was added to the lipid phase, and the formulation was emulsified by sonication using an ultrasonicator equipped with a 3.2 mm microprobe tip, operating at an amplitude of 240 μm and a frequency of 20 kHz for 2 minutes at room temperature. The compound concentration in all formulations was verified using HPLC-MS-MS.

[0232] Male Sprague-Dawley (SD) rats were selected for lymphatic transport studies in which the drug was mycophenolic acid (MPA), sertraline (SER), or buprenorphine (BUP). Female SD rats were selected for studies in which the drug was testosterone. This was to eliminate the possibility that the relatively high and fluctuating endogenous testosterone levels in male rats might interfere with the quantification of exogenously administered testosterone. Rats (220–320 g) were maintained on standard chow and fasted overnight prior to the experiment, with water available ad libitum. Anesthetized rats were placed on a heating pad at 37°C, and the duodenum (for administration of formulations and hydration), mesenteric lymphatic duct (for lymph collection), and carotid artery (for blood collection) were cannulated as previously described (Edwards et al. Advanced Drug Delivery Reviews 2001, 50(1), 45–60). After surgery, rats were rehydrated for 0.5 hours by intraduodenally infusing normal saline at 2.8 mL / hour. The lipid formulation was infused intraduodenally at 2.8 mL / hour for 2 hours, followed by normal saline at 2.8 mL / hour for the remainder of the experiment. Lymph was continuously collected for up to 8 hours into preweighed Eppendorf tubes containing 10 μL of 1,000 IU / mL heparin. Collection tubes were changed every hour, and lymph flow was measured gravimetrically. Aliquots of hourly lymph samples were stored at -80°C before assay.

[0233] Drug concentrations in lymph are expressed as total drug and include free drug and drug associated with different glycerides, which are assayed by hydrolyzing the lymph (to liberate the drug from any re-esterified glycerides) before assessing free drug.

[0234] Transport of compound into lymph during each hourly collection period was calculated from the product of the collected lymph volume and the measured lymph concentration.

[0235] As shown in Figure 1 and Table 4, the lymphatic transport of compound 12, which has an acetal self-immolative group (ASI), compound 15, which has a trimethyl lock (TML) self-immolative group, and compound 17, which has a (4-carbon) inverted ester self-immolative group (FSI-4), was 1.9%, 3.2%, and 5.2% (of the administered dose), respectively. This is lower than the corresponding linear testosterone-succinate-TG (compound 21, 13.4%), and compound 12 is reduced to the level of the currently marketed testosterone prodrug, testosterone undecanoate (TU). The reduced lymphatic transport of the prodrugs containing the self-immolative group is likely due to poor stability of the monoglyceride form of the prodrug in the gastrointestinal tract (as described in Example 11 below and shown in Figure 9), or potentially due to reduced efficiency of re-esterification of the monoglyceride form in enterocytes. The order of stability of the monoglyceride forms of Compound 21, Compound 12, and Compound 17 (i.e., Compound 21-monoglyceride > Compound 17-monoglyceride > Compound 12-monoglyceride) is consistent with the order of lymphatic transport of these three prodrugs (i.e., Compound 21 > Compound 17 > Compound 12).

[0236] The inclusion of a methyl group at the alpha or beta carbon relative to the glyceride unit enhances the stability of the monoglyceride intermediate of the prodrug. For example, to address stability issues with compound 12, a methyl protecting group was included to form compounds 13 and 14 (these compounds were used as a mixture of compounds 13 and 14). As evident from Figure 1 and Table 4, the inclusion of a methyl protecting group significantly enhanced the lymphatic transport of compounds 13 and 14 compared to compound 12. This is consistent with their enhanced stability under GI digestion conditions (see Figure 9). The lymphatic transport of compound 19 (containing a 5-carbon inverted ester self-immolative group [FSI-5] with a methyl branch) was 9.6% (of the administered dose). This is also higher than the lymphatic transport of a similar compound, compound 17 (containing an FSI-4 self-immolative group), which lacks a methyl group.

[0237] [Table 4]

[0238] As shown in Figure 4 and Table 5, the lymphatic transport of compound 3, which contains a TML self-immolative group, was 21.5% (of the administered dose). In contrast, the lymphatic transport of the parent drug sertraline (SER) after administration of the parent drug SER.HCl was only 0.05% (of the administered dose).

[0239] [Table 5]

[0240] As shown in Figure 6 and Table 4, the lymphatic transport of compound 6 with a TML self-immolative group and compound 7 with a (5-carbon) inverted ester self-immolative group (FSI-5) was 10.8% and 24.5% (of the administered dose). In contrast, after administration of buprenorphine (BUP), the lymphatic transport of the parent drug BUP was extremely low, at only 0.01% (of the administered dose).

[0241] [Table 6]

[0242] As shown in Figure 8 and Table 7, the lymphatic transport of compound 10, which has an acetal self-immolative (ASI) group, and compound 11, which has a methyl acetal self-immolative (MASI) group, was 0.35% and 1.22% (of the administered dose). In contrast, the lymphatic transport of the parent drug, mycophenolic acid (MPA), after administration of MPA was only 0.17% (of the administered dose).

[0243] [Table 7]

[0244] Example 10. Pharmacokinetic (PK) studies in rats To assess the oral bioavailability of the compounds of the present invention, pharmacokinetic studies were performed using the following procedure. The day before drug administration, female (for testosterone-related studies) and male (for SER and BUP-related studies) Sprague-Dawley rats (220–320 g) were anesthetized and their carotid arteries were cannulated. The rats were then allowed to regain consciousness and fasted overnight prior to the start of the experiment, with free access to water. The following morning, formulations containing the parent compound or prodrug were administered via oral gavage. Blood samples were collected from the carotid artery cannula from 5 minutes before dosing until 24 hours after dosing, and plasma was separated by centrifugation at 5000 rpm for 5 minutes. Between blood sample collection times, rats had free access to water but remained fasted for an additional 8 hours after drug administration. Plasma samples were stored at -80°C before assay by HPLC-MS-MS. In this case, samples were assayed for free drug (i.e., drug not associated with glycerides) and were not hydrolyzed prior to assay (as was the case for lymph samples). Thus, the data reflect drug released from re-esterified drug-glyceride complexes in the systemic circulation after transport to the lymph.

[0245] As previously explained, triglyceride prodrugs employing short linkers (e.g., succinic acid, compound 21) between the drug testosterone and the glyceride unit are limited by poor drug release in the systemic circulation (see Scriba, GKE, Arch. Pharm. (Weinheim). 1995, 328, (3), 271-276 and Scriba, GKE et al., J. Pharm. Pharmacol. 1995, 47, (11), 945-948). As shown in Figure 2 and Table 8 below, the addition of a self-immolative group to the linker increases the systemic exposure of the drug, even when combined with the previously rejected succinic acid linker, thereby bypassing first-pass metabolism of the drug and increasing its oral bioavailability.

[0246] Figure 2 shows dose-normalized testosterone plasma concentrations after oral gavage of testosterone formulations to conscious, carotid-cannulated female SD rats. The formulations contained approximately 2 mg of a compound of the invention containing 1 mg of TU or TST, or testosterone, dispersed in 40 mg of oleic acid, 25 mg of Tween 80, and 2 ml of PBS. Doses are normalized to a 2 mg / kg equivalent dose of testosterone. Data are shown as mean ± SEM. The inset shows the dose-normalized plasma AUC of testosterone in bar graph format. 0-24h (nmol x hr / L).

[0247] Table 8 shows the pharmacokinetic parameters of parent testosterone after oral administration of compounds 12–20 or 21. In all cases where the short-chain linker did not contain any methyl groups (compounds 12, 15–18, or 20), the systemic exposure of testosterone was greater than that of compound 21 (approximately 1.4–18-fold increase) or the commercial product TU (approximately 4–54-fold increase). Although the lymphatic transport of compounds 12 and 17 was relatively low (Figure 1 and Table 4), the systemic exposure of parent testosterone was still much higher after oral administration of both prodrugs. This suggests that the self-immolative group facilitates the conversion of the prodrugs to the parent drug in the systemic circulation.

[0248] [Table 8]

[0249] The utility of prodrugs can be further enhanced by enhancing their lymphatic transport. The inclusion of a methyl group at either end of the short-chain linker, either alpha or beta to the ester, can enhance the stability of the monoglyceride intermediate of the prodrug and thus enhance lymphatic transport. The in vitro digestion and lymphatic transport results of compound 12 relative to compounds 13 / 14 (see Figure 9, Figure 1, and Table 4) support this suggestion, demonstrating significantly enhanced stability of compound 13 / 14 under simulated intestinal conditions. Consistent with the increased lymphatic transport of compound 13 / 14 and the increased potential of the self-immolative group to promote systemic release of testosterone from the prodrug, the systemic exposure of testosterone following administration of compound 13 / 14 or compound 13 alone was approximately 13-fold higher than compound 12 and 95-97-fold higher than TU (Table 8). Additionally, in the case of the FSI prodrug, the inclusion of a methyl group in the short-chain linker also enhanced the bioavailability of testosterone. Thus, the systemic exposure of testosterone after administration of compound 19 was 1.9-fold higher than that of compound 18 and 1.9-fold higher than that of TU. is also 105 times higher (Table 8).

[0250] Figure 5 shows dose-normalized SER plasma concentrations after oral gavage of formulations to conscious, carotid-cannulated male SD rats. In the SER parent drug control group, the formulation contained 0.7 mg of SER.HCl dissolved in 2 ml of water. The prodrug formulation contained 2 mg of a compound of the invention containing SER dispersed in 40 mg oleic acid, 25 mg Tween 80, and 2 ml of PBS. Doses are normalized to a 2 mg / kg equivalent dose of SER. Data are shown as mean ± SEM. The inset shows the dose-normalized plasma AUC of SER in bar graph form. 0-24h (nmol x hr / L).

[0251] The pharmacokinetic parameters of SER after administration of SER.HCl, compounds 1, 2, and 3 are shown in Table 9. In all cases, the systemic exposure of SER after administration of the prodrug was greater (2- to 3-fold increase) than that of SER.HCl. This suggests that the prodrug is lymphatically transported (as exemplified by compound 3 in Figure 4 and Table 5) and that the self-immolative group facilitates conversion of the prodrug to the parent drug in the systemic circulation.

[0252] [Table 9]

[0253] Figure 7 shows the dose-normalized BUP plasma concentrations after oral gavage of formulations to conscious, carotid-cannulated male SD rats. In the BUP parent drug control group, the formulation contained 0.02 mg of BUP dissolved in 2 ml of 0.1% aqueous acetic acid. The prodrug formulation contained 40 mg of oleic acid, 25 mg of Tween 100, and 100 mg of ethanol. The samples contained 0.05 mg of a compound of the invention containing BUP dispersed in 80 ml of PBS and 2 ml of PBS. Doses are normalized to an equivalent dose of 0.06 mg / kg of BUP. Data are shown as mean ± SEM. The inset shows the dose-normalized plasma AUC of BUP in bar graph form. 0-6h (nmol x hr / L).

[0254] The pharmacokinetic parameters of BUP after administration of BUP, compounds 5, 6, and 7 are shown in Table 10. In all cases, the systemic exposure of BUP after administration of the compounds of the invention was greater than that of BUP (7- to 14-fold increase). This suggests that the prodrugs are lymphatically transported (as exemplified by compounds 6 and 7 in Figure 6 and Table 6) and that the self-immolative group facilitates the conversion of the prodrug to the parent drug in the systemic circulation. This suggests that:

[0255] [Table 10]

[0256] Example 11. In vitro hydrolysis of compounds with porcine pancreatic lipase In vitro hydrolysis of testosterone prodrugs was carried out via incubation with porcine pancreatic lipase. Briefly, prior to the hydrolysis experiment, a pancreatic lipase solution was prepared by dispersing 1 g of porcine pancreatin in 5 ml of lipolysis buffer. The suspension was mixed well and centrifuged at 3500 rpm for 15 minutes at 5°C to obtain a supernatant. A volume of 1000 ml of lipolysis buffer was prepared using 0.474 g of Tris-maleic acid (2 mM), 0.206 g of CaCl2.HO (1.4 mM), and 8.775 g of NaCl (150 mM), adjusted to pH 6.5 with NaOH. To evaluate the potential for prodrug hydrolysis in the intestine, 20 μl of prodrug solution (1 mg / ml, dissolved in acetonitrile), 900 μl of simulated intestinal micelle solution (prepared with 0.783 g NaTDC (3 mM) and 0.291 g phosphatidylcholine (0.75 mM) in 500 ml of lipolysis buffer), and 100 μl of enzyme solution were incubated at 37°C. 20 μl samples of the incubation solution were taken after 0, 5, 10, 15, 30, 60, 90, 120, and 180 min of incubation and added to 180 μl of ACN to stop lipolysis. The mixture was vortexed and centrifuged at 5000 rpm for 5 min to precipitate proteins before analysis. The supernatant was analyzed by HPLC-MS for residual compound concentration and potential products of compound hydrolysis.

[0257] Upon incubation with digestive enzymes, the monoglyceride forms of the prodrugs are formed very rapidly. Therefore, the stability in simulated intestinal conditions is better assessed by the stability of the monoglyceride form generated by the initial digestive process. The monoglyceride form must remain intact to be absorbed and re-esterified in the enterocytes before entering the lymphatics. Figure 9 shows the stability profiles of the monoglyceride forms of Compound 12, Compounds 15-17, and Compound 20 during in vitro incubation with freshly prepared porcine pancreatic lipase (n=2-3 for each group). ) is compared to that of the non-self-immolative containing compound 21 (n=3). This data shows that the inclusion of an acetal (ASI), inverted ester (FSI), carboxy(methyl acetal) (CMSI), or p-hydroxybenzylcarbonyl (PHB) self-immolative group results in significantly reduced luminal stability of the MG form of the prodrug. However, the trimethyl-locked self-immolative group does not appear to affect luminal stability, as the monoglyceride form of compound 15 is as stable as the monoglyceride form of compound 21.

[0258] Figure 9 also provides evidence of the ability of methyl substitution to improve the luminal stability of testosterone prodrugs containing acetal self-immolative linkers (acetal groups normally reduce luminal stability). As shown in the figure, the monoglyceride form of compound 12 was unstable in an in vitro lipolysis assay. In contrast, the mixture of alpha- and beta-methyl-substituted compounds 13 / 14 was much more stable. The enhanced stability of the methyl-substituted ASI prodrugs most likely accounts for the significant increase in in vitro lymphatic transport and the increase in testosterone exposure in the systemic circulation after oral administration.

[0259] Example 12. In vitro release of MPA from prodrug in lymph supplemented with lipoprotein lipase To investigate the release of free MPA from TG prodrugs in lymphatic vessels (the active site of MPA is located in lymphocytes, which are abundant in the lymphatic system), MPA prodrugs were incubated with rat lymph supplemented with lipoprotein lipase (LPL, 200 units / ml). LPL is a key enzyme required for the hydrolysis of lipoprotein-associated TG under normal physiological conditions and is therefore expected to be an important contributor to the lipolysis of re-esterified drug-TG constructs in plasma, largely due to the liberation of FAs at the sn-1 and sn-3 positions of the TG-mimetic prior to drug release from the 2' position via esterase hydrolysis. LPL is tethered to lymphocytes or lymphatic / vascular endothelial cells under physiological conditions. Therefore, in this in vitro study, rat lymph was supplemented with LPL to better reflect the in vitro situation. To initiate hydrolysis, 10 μl of LPL solution (10,000 units / ml) was added to a mixture of 10 μl of prodrug solution (1 mg / ml, dissolved in acetonitrile) and 500 μl of blank Sprague-Dawley rat lymph. The solution was incubated at 37°C. Samples (20 μl) of the incubation solution were taken at 0, 5, 10, 15, 30, 60, 90, 120, and 180 minutes after incubation and added to 980 μl of 9:1 (v / v) ACN-water to stop lipolysis. The mixture was vortexed and centrifuged at 4500 g for 5 minutes to precipitate proteins before analysis. The supernatant was analyzed for MPA concentration by HPLC-MS / MS.

[0260] As shown in Figure 10 (n=1 for each prodrug), the release of pharmacologically active MPA from compounds 10 and 11 was very rapid when incubated with rat lymph supplemented with LPL. The release rates of parent MPA from these two SI-group-containing prodrugs were much higher than that from MPA-TG, a prodrug lacking an SI group. This data indicates that the inclusion of an acetal (ASI) or methyl acetal (MASI) self-immolative group results in significantly enhanced release of the parent compound, thus providing an opportunity for targeted delivery of therapeutic agents to active sites in the lymphatic system.

Claims

1. Compounds of formula (I): 【Chemical 1】 [In the formula, R １ and R ２ are independently H or C ２ ~C ２８ represents a residue of a fatty acid, -X- is selected from -O-, -NH-, and -S-; 【Chemistry 2】 represents the residue of a drug, -L- is —OC(O)— or —X′—; When -L- is -OC(O)-, -Y- is an optionally substituted -C １ ~C ２０ AlkylC(O)OCH ２ -, -C ２ ~C ２０ Alkenyl C(O)OCH ２ - or -C ２ ~C ２０ Alkynyl C(O)OCH ２ - group, wherein one or more of the carbon atoms of the alkyl, alkenyl, or alkynyl group is selected from the group consisting of NH, S, O, C ５ ~C ８ an aromatic or aliphatic cyclic group, or C ５ ~C ８ may be substituted with an aromatic or aliphatic heterocyclic group (provided that the alkyl, alkenyl, or alkynyl group is not a straight-chain C ２０ provided that the length does not exceed the length of the alkyl group; or When -L- is -X'-, -Y- is an optionally substituted -C １ ~C ２ Alkyl C(O)R ３ - group, or -C ２ Alkenyl C(O)R ３ - or -C ２ Alkynyl C(O)R ３ represents a - group, R ３ is a self-immolative group, X' is O, S, N(R ４ ), or N(H)S(O) ２ and R ４ is H or C １ ~C ４ alkyl, or Pharmaceutically acceptable salts thereof.

2. R ３ teeth, 【Chemistry 3】 [In the formula, 【Chemistry 4】 indicates the point of attachment to the drug residue and the -Y- group.

2. The compound of claim 1 selected from:

3. The compound of claim 1 represented by formula (II): 【Chemistry 5】 [In the formula, R １ , R ２ and -X- are as defined in claim 1; R ３ is a self-immolative group, 【Chemistry 6】 represents the residue of a drug, -L- is -X'-; X' is O, S, N(R ４ ), or N(H)S(O) ２ and R ４ is H or C １ ~C ４ is alkyl, R ５ is hydrogen and C １ ~C ４ alkyl; or Pharmaceutically acceptable salts thereof.

4. The compound of claim 1 represented by formula (III): 【Chemistry 7】 [In the formula, R １ , R ２ and -X- is as defined in claim 3; R ３ is a self-immolative group, 【Chemistry 8】 represents the residue of a drug, -L- is -X'-; X' is O, S, N(R ４ ), or N(H)S(O) ２ and R ４ is H or C １ ~C ４ is alkyl, R ５ and R ６ is hydrogen and C １ ~C ４ alkyl; or Pharmaceutically acceptable salts thereof.

5. The compound of claim 1 represented by formula (IV): 【Chemistry 9】 [In the formula, R １ , R ２ and -X- is as defined in claim 3; 【Chemistry 10】 represents the residue of a drug, R ５ and R ６ is hydrogen and C １ ~C ４ alkyl; R ７ is hydrogen or C １ ~C ４ is alkyl, n is 0 to 18; or Pharmaceutically acceptable salts thereof.

6. 6. The compound of any one of claims 1 to 5, wherein the agent exhibits a first-pass metabolism rate of greater than 50% or has a highly variable first-pass metabolism rate after oral administration.

7. 7. The compound of any one of claims 1 to 6, wherein the agent is selected from testosterone, mycophenolic acid, estrogen (estrogens), opiates such as morphine, tetrahydrocannabinol, cannabidiol, metoprolol, raloxifene, alphaxolone, statins such as atorvastatin, buprenorphine, pentazocine, propranolol, L-DOPA, lidocaine, chlorpromazine, amitriptyline, nortriptyline, oxprenolol, labetalol, salbutamol, epitiostanol, melphalan, or lovastatin.

8. 8. The compound of claim 7, wherein the agent is testosterone and the compound is represented by formula (V): 【Chemistry 11】 [In the formula, R １ , R ２ and -X- are as defined in claim 1; R ５ and R ６ is hydrogen and C １ ~C ４ alkyl; R ３ is a self-immolative group; or Pharmaceutically acceptable salts thereof.

9. R ５ is methyl, and R ６ 9. The compound of claim 4, wherein is hydrogen.

10. R ５ is hydrogen, and R ６ 9. The compound of claim 4, wherein is methyl.

11. 11. The compound of claim 1, wherein X is oxygen.

12. R １ and R ２ is a residue of palmitic acid.

13. A method for treating or preventing a disease or disorder in which increased testosterone levels are beneficial, comprising administering to a subject in need thereof a therapeutically effective amount of a compound described in any one of claims 8 to 12.

14. 14. The method of claim 13, wherein the disease or disorder is hypogonadism, anemia due to bone marrow failure, anemia due to renal failure, chronic respiratory failure, chronic heart failure, a steroid-dependent autoimmune disorder, AIDS wasting, hereditary angioedema or urticaria, end-stage breast cancer, or menopause.

15. A method for enhancing lymphatic transport and systemic release of a drug, comprising: adding to a pharmaceutical compound a prodrug moiety of formula (VI): 【Chemistry 12】 [In the formula, R １ and R ２ are independently H or C ２ ~C ２８ represents a residue of a fatty acid, -X- is selected from -O-, -NH-, and -S-; -Y- is an optionally substituted -C １ ~C ２ Alkyl C(O)R ３ - group, or -C ２ Alkenyl C(O)R ３ - or -C ２ Alkynyl C(O)R ３ represents a - group, R ３ is a self-immolative group, 【Chemistry 13】 indicates the point at which the linker is conjugated to the pharmaceutically active agent; or conjugating a pharmaceutically acceptable salt thereof.

16. R ３ teeth, 【Chemistry 14】 [In the formula, 【Chemistry 15】 indicates the point of attachment to the drug residue and the -Y- group.

16. The method of claim 15, wherein the compound is selected from the group consisting of:

17. 17. The method of any one of claims 13 to 16, wherein the compound is administered orally with food to promote transport to the intestinal lymph.

18. 18. The method of any one of claims 13 to 17, wherein the compound is orally co-administered with a lipid-based formulation to facilitate transport to the intestinal lymph.

19. 17. The method of any one of claims 13 to 16, wherein the compound is orally co-administered with an enzyme inhibitor.

20. 6. The compound of any one of claims 1 to 5, selected to facilitate targeted delivery of the agent within the lymphatic system.

21. Such drugs include nonsteroidal anti-inflammatory drugs (NSAIDS, e.g., aspirin, ibuprofen, naproxen), COX-2 inhibitors (e.g., celecoxib), corticosteroid anti-inflammatory drugs (e.g., prednisolone, dexamethasone), antimalarials (e.g., hydroxychloroquine), nitrosoureas, methotrexate, dactinomycin, anthracyclines (e.g., daunorubicin), mitomycin C, bleomycin, mithramycin, drugs acting on immunophilins (e.g., cyclosporine, tacrolimus, sirolimus), sulfasalazine, leflunomide, mycophenolate, opioids, fingolimod, myriocin, chlorambucil, doxorubicin, nelarabine, cortisone, dexamethasone, prednisone, pralatrexate, vinblastine, bortezomib, nelarabine, daunorubicin hydrochloride, clofarabine, cytarabine, dasatinib, imatinib Mesylate, ponatinib hydrochloride, vincristine sulfate, bendamustine hydrochloride, fludarabine phosphate, bosutinib, nilotinib, omacetaxine mepesuccinate, capecitabine, paclitaxel, gemcitabine, fulvestrant, tamoxifen, lapatinib, toremifene, ixabepilone, eribulin, albendazole, ivermectin, diethylcarbamazine, albendazole, doxycycline, closantel, mala 21. The compound of claim 20, selected from viroc, enfuvirtide, deoxythymidine, zidovudine, stavudine, didanosine, zalcitabine, abacavir, lamivudine, emtricitabine, tenofovir, delavirdine, rilpivirine, raltegravir, elvitegravir, lopinavir, indinavir, nelfinavir, amprenavir, ritonavir, acyclovir, immunosuppressants such as mycophenolic acid, and pharmaceutically active peptides.

22. 22. A pharmaceutical composition comprising a therapeutically effective amount of a compound of any one of claims 1 to 12, 20, or 21, or a pharmaceutically acceptable salt thereof, together with at least one pharmaceutically acceptable carrier or diluent.

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