Rapamycin analogs and uses thereof

Abstract

The present invention provides compounds, compositions thereof, and methods of using same.

Description

[Technical Field]

[0001] (CROSS-REFERENCE TO RELATED APPLICATIONS) This application claims the benefit under 35 U.S.C. §119(e) to U.S. Provisional Patent Application No. 63 / 140,523, filed January 22, 2021, and U.S. Provisional Patent Application No. 63 / 202,524, filed June 15, 2021, the disclosures of each of which are incorporated herein by reference in their entireties.

[0002] FIELD OF THE INVENTION The present invention relates to compounds and methods useful for modulating mTORC1 activity. The present invention also provides pharmaceutically acceptable compositions comprising the provided compounds of the present invention, and methods of using such compositions in the treatment of various disorders. [Background technology]

[0003] mTOR complex 1 (mTORC1) positively regulates cell growth and proliferation by promoting many anabolic processes, including the biosynthesis of proteins, lipids, and organelles, and by limiting catabolic processes such as autophagy. Much of our knowledge of mTORC1 function comes from the use of the bacterial macrolide rapamycin. Upon entering the cell, rapamycin binds to the 12-kDa FK506-binding protein of 12 (FKBP12), interacting with the FKBP12-rapamycin binding domain (FRB) of mTOR, thus inhibiting mTORC1 function (Guertin, DA & Sabatini, DM Cancer Cell 12(1):9-22 (2007)). In contrast to its effects on mTORC1, FKBP12-rapamycin cannot physically interact with or acutely inhibit mTOR complex 2 (mTORC2) (Janinto, E. et al., Nat. Cell Bio., 6(11):1122-8(2004); Sarbassov, D.D. et al., Curr. Biol. 14(14):1296-302(2004)). Based on these observations, mTORC1 and mTORC2 have been characterized as rapamycin-sensitive and rapamycin-insensitive complexes, respectively. However, this paradigm may not be entirely accurate, as chronic rapamycin treatment can, in some cases, inhibit the activity of mTORC2 by blocking its assembly (Sarbassov, D.D. et al., Mol. Cell, 22(2):159-68(2006)).In addition, recent reports suggest that key functions of mTORC1 are resistant to inhibition by rapamycin (Choo, AY et al., Proc. Natl. Acad. Sci., 105(45):17414-9 (2008); Feldman, ME et al., PLoS Biol., 7(2):e38 (2009); Garcia-Martinez, J Met al., Biochem J., 421(1):29-42 (2009); Thoreen, CC et al., J. Biol. Chem., 284(12):8023-32 (2009)). Therefore, selective inhibition of mTORC1 may enable the treatment of diseases involving dysregulated protein synthesis and cellular metabolism. Furthermore, this detailed understanding of the regulation of mTORC1 activation pathways may enable the discovery of new strategies for modulating abnormal disease processes by modulating mTORC1 activity across the spectrum of its functions.

[0004] Many diseases are associated with abnormal cellular responses triggered by such events, including, but not limited to, autoimmune diseases, inflammatory diseases, bone diseases, metabolic diseases, neurological and neurodegenerative diseases, cancer, cardiovascular diseases, allergies and asthma, Alzheimer's disease, and hormone-related diseases.

[0005] The mechanistic target of rapamycin complex 1 (mTORC1) is a master growth regulator that senses diverse environmental factors, such as growth factors, cellular stress, and nutrient and energy levels. Upon activation, mTORC1 phosphorylates substrates to enhance anabolic processes, such as mRNA translation and lipid synthesis, and limit catabolic processes, such as autophagy. Dysregulation of mTORC1 occurs in a wide range of diseases, including diabetes, epilepsy, neurodegeneration, immune responses, suppressed skeletal muscle growth, and, particularly, cancer (Howell, JJ et al., Biochem. Soc. Trans., 41:906-12 (2013); Kim, S. Get al., Molecular and Cells, 35(6):463-73 (2013); Laplante, M. & Sabatini, DM, Cell, 149(2):274-93 (2012)).

[0006] Rapamycin was first discovered as an antifungal metabolite produced by Streptomyces hygroscopicus in soil samples from Easter Island. It was subsequently found to have immunosuppressive and antiproliferative properties in mammalian cells, sparking interest in identifying its mode of action. Rapamycin was shown to be a potent inhibitor of S6K1 phosphorylation. Concurrently, the target of rapamycin (TOR) was identified in yeast and mammalian cells. Rapamycin forms a gain-of-function complex with the 12 kDa FK506-binding protein (FKBP12), which specifically binds to and acts as an allosteric inhibitor of mammalian TOR (mTOR, also known as mechanistic TOR) complex 1 (mTORC1).

[0007] Biochemical and genetic analyses have demonstrated that mTOR exists in two functionally distinct complexes. The core components of mTORC1 consist of mTOR, mammalian lethal with sec-13 protein 8 (mLST8), and regulatory-associated protein of TOR (Raptor). Additional components include DEP-domain-containing mTOR-interacting protein (DEPTOR) and 40 kDa proline-rich Akt substrate 40 (PRAS40).

[0008] The mTOR complex 2 (mTORC2) core consists of mTOR, the rapamycin-insensitive companion of mTOR (Rictor), stress-activated protein kinase-interacting protein 1 (mSIN1), and mLST8. The proteins observed together with rictor1 / 2 (protor1 / 2) and DEPTOR are additional regulatory components. S6 kinase 1 (S6K1) and eukaryotic inhibitor of eIF4E-binding protein 1 (4E-BP1) are two well-characterized substrates of mTORC1, while AKT is a well-characterized substrate of mTORC2 (Li, J. et al., Cell Met., 19(3):373-9 (2014)).

[0009] Because FKBP12-rapamycin does not bind to mTORC2, rapamycin was initially thought to inhibit only mTORC1 (Sarbassov, DD et al., Curr. Biol., 14(14):1296-302 (2004)). However, in 2006, it was shown that rapamycin suppresses the assembly and function of mTORC2 and inhibits pAkt (Sarbassov, DD et al., Molecular Cell, 22(2):159-68 (2006)). The effects of rapamycin on the phosphorylation of S473 of Akt (an mTORC2 substrate) and T389 of S6K1 (an mTORC1 substrate) were compared in multiple cell lines. In PC3, HEK-293T, HeLa, and H460 cells, treatment with rapamycin for 1 or 24 hours inhibited S6K1 phosphorylation, consistent with mTORC1 inhibition. Selective inhibition of S6K1 by rapamycin should result in increased Akt phosphorylation, which was indeed reported in HeLa cells. However, in PC3 cells, the drug strongly reduced Akt phosphorylation, suggesting that rapamycin is not selective in this cell line. Partial inhibition of pAKT was observed in HEK-293T cells. In approximately one-third of cell lines, rapamycin caused strong or partial inhibition of Akt phosphorylation, whereas in other cell lines, the drug did not affect or increase Akt phosphorylation. Inhibition of pAKT after 24 hours has also been observed in primary and non-transformed cell lines, including endothelial and muscle cells. Furthermore, mice treated daily with rapamycin for 1 week showed decreased Akt phosphorylation in the thymus, adipose tissue, heart, and lung, indicating that the drug inhibits pAkt in vivo. These findings indicate that rapamycin-mediated inhibition of Akt phosphorylation is general and occurs in normal and cancer cell lines, as well as in vivo.

[0010] concluded that rapamycin and its analogs (CCI779, RAD001 (also known as everolimus), and AP23573) can inhibit mTORC2 function in certain cell lines and tissues. Rapamycin-mediated Akt inhibition may help explain the drug's side effects. For example, rapamycin potently inhibits Akt phosphorylation in adipose tissue, a tissue type in which insulin-stimulated Akt activity plays a key role in suppressing lipolysis. Rapamycin-mediated Akt inhibition in adipocytes may maintain high lipolysis even in the presence of insulin, leading to the accumulation of plasma free fatty acids (which can be used by the liver to generate triglycerides), providing a molecular mechanism for the hyperlipidemia commonly seen in patients treated with rapamycin.

[0011] Pereira et al. (Mol Cell Endocrinol., 355(1):96-105(2012)) investigated the effects of rapamycin on glucose uptake and insulin signaling proteins in adipocytes obtained from fat biopsies of human donors. At therapeutic concentrations (0.01 μM), rapamycin reduced the phosphorylation of Ser473 of AKT (PKB) and reduced glucose uptake in human adipocytes through impaired insulin signaling.

[0012] Lamming et al. (Science., 335(6076):1638-1643(2012)) showed that rapamycin disrupts mTORC2 in vivo and that mTORC2 is required for insulin-mediated suppression of hepatic gluconeogenesis.

[0013] Similar results have been shown in humans. Di Paolo et al. published similar findings in humans (JASN, 17(8):2236-2244(2006)). The primary goal of their study was to determine the effect of chronic exposure to rapamycin on AKT activation, given its important role in regulating cell proliferation and survival and in cellular responses to nutrients and growth factors. They found that mTOR inhibition was associated with a significant downregulation of basal and insulin-induced AKT phosphorylation. AKT is primarily involved in many of the metabolic effects of insulin, and therefore, they concluded that suppression of AKT activation significantly correlates with increased insulin resistance in renal transplant recipients.

[0014] Kennedy et al. recently reviewed the role of mTORC1 and mTORC2 in metabolism and aging (Cell Metab., 23(6):990-1003(2016)). Summary of the Invention

[0015] Therefore, there remains a need to provide potent mTORC1-specific inhibitors with improved safety and tolerability due to the lack of direct mTORC2 inhibition.

[0016] It has now been found that compounds of the present invention and pharmaceutically acceptable compositions thereof are effective as inhibitors (mTORC1 inhibitors). Such compounds have the general formula I:

[0017] [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 , R 3 , R 3’ , R 4 , R 5 , R 5’ , R 6 , L 1 , L 2 , X, X 1 , X 2 , and X 3Each of is as defined and described herein.

[0018] The compounds of the present invention and pharmaceutically acceptable compositions thereof are useful for treating a variety of diseases, disorders, or conditions associated with mTORC1, including those described herein. [Brief explanation of the drawings]

[0019] [Figure 1] A comparison of Western blots performed after treating PC3 cells with rapamycin, I-59, I-57, and I-55 for 24 hours is shown. [Figure 2] A comparison of Western blots performed after treating PC3 cells with rapamycin, I-69, I-66, I-64, and I-62 for 24 hours is shown. [Figure 3] A comparison of Western blots performed after treating PC3 cells with rapamycin, I-85, I-97, and I-83 for 24 hours is shown. [Figure 4] A comparison of Western blots performed after treating PC3 cells with rapamycin, I-34, I-49, and I-31 for 24 hours is shown. [Figure 5] A comparison of Western blots performed after treating PC3 cells with rapamycin, I-37, I-43, and I-40 for 24 hours is shown. [Figure 6] A comparison of Western blots performed after treating PC3 cells with rapamycin and I-14 for 24 hours is shown. [Figure 7] A comparison of Western blots performed after treating PC3 cells with rapamycin, I-4, I-27, and I-47 for 24 hours is shown. [Figure 8] A comparison of Western blots performed after treating PC3 cells with rapamycin, I-9, and I-21 for 24 hours is shown. [Figure 9] A comparison of Western blots performed after treating PC3 cells with rapamycin, I-18, and I-45 for 24 hours is shown. DETAILED DESCRIPTION OF THE INVENTION

[0020] 1. General Description of Certain Embodiments of the Invention: Surprisingly, the provided compounds have been found to inhibit mTORC1 but not mTORC2 (as measured by their effects on pAKT) over extended periods (e.g., 8 hours, 24 hours, 30 hours, and 48 hours). This novel activity is due to the presence of a sufficiently large group at the C-7 position of rapamycin and its analogs. Small substituents at this position, such as OMe (as found in rapamycin), OEt, and OBn, do not confer selectivity for mTORC2 at 24 hours. Medium-length groups, such as OCH2CH2OH or OCH2CH2CH2OH, show partial selectivity for mTORC2 at 24 hours, but still exhibit some inhibition. In comparison, larger groups, such as those of the present invention, provide significant selectivity for mTORC2 as measured by effects on pAKT.

[0021] The position of this substitution is also important to the observed selectivity: the introduction of a larger substituent at the C-43 position, for example, does not result in the unique selectivity profile claimed in this application.

[0022] For clarity, the structure of rapamycin is reproduced below with the C-7 and C-43 positions indicated.

[0023] [ka]

[0024] In some embodiments, the present invention provides novel rapamycin analogs that are potent inhibitors of mTORC1 as measured by pS6K. Unlike rapamycin and everolimus, these compounds do not inhibit pAKT at longer time points (e.g., 24 hours and 48 hours). These compounds also exhibit improved solubility and improved pharmacokinetics compared to rapamycin.

[0025] The activity of the compounds used in the present invention as inhibitors of mTORC1 can be assayed in vitro, in vivo, or in cell lines. In vitro assays include assays that determine the inhibition of mTORC1. Detailed conditions for assaying the compounds used in the present invention as inhibitors of mTORC1 are well known to those skilled in the art. Such methods are described in detail by Liu et al., Cancer Research, 73(8):2574-86 (2013) and Liu et al., J. Biological Chemistry 287(13):9742-52 (2012).

[0026] In certain embodiments, the present invention provides a compound of formula I,

[0027] [ka] During the ceremony, X and X 3 are independently a covalent bond, -CR2-, -NR-, -NRCO-, -NRCO2-, -NRCONR-, -NRSO2-, -O-, -S-, or -SO2NR-; L 1 is a covalent bond or C 1~30 a divalent linear or branched saturated or unsaturated hydrocarbon chain in which 1 to 10 methylene units are independently and optionally replaced by -Cy1-, -O-, -S-, -S(O)2-, -C(O)-, -C(S)-, -C(R)2-, -CH(R)-, -CF2-, -P(O)(R)-, -Si(R)2-, -Si(OR)(R)-, or -NR-; each -Cy1- is independently an optionally substituted bivalent ring selected from phenylene, 4- to 7-membered saturated or partially unsaturated heterocyclylene having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and 5- to 6-membered heteroarylene having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; Each R is independently hydrogen or C 1~6 an optionally substituted group selected from aliphatic, 3- to 8-membered saturated or partially unsaturated monocyclic carbocycle, phenyl, 4- to 7-membered saturated or partially unsaturated heterocycle having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and 5- to 6-membered heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; or two R groups on the same atom, taken together with their intervening atoms, form a 4-7 membered saturated, partially unsaturated, or aryl ring having, in addition to the same atom to which they are attached, 0-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; L 2 optionally replaced by C 1~6 a divalent linear or branched saturated or unsaturated hydrocarbon chain, wherein one to two methylene units of said chain are independently and optionally replaced with -Cy1-; R 1 and R 2 are independently hydrogen, halogen, -OR, -CN, -NO2, -NR2, -NR(C 1~6 haloalkyl), -NRCO, -NRCO2R, -NRCONR2, -NRSO2R, -SR, -SO2NR2, or C 1~6an optionally substituted group selected from an aliphatic, a 3- to 8-membered saturated or partially unsaturated monocyclic carbocycle, a phenyl, an 8- to 10-membered bicyclic aromatic carbocycle, a 4- to 8-membered saturated or partially unsaturated monocyclic heterocycle having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, and sulfur, a 7- to 10-membered saturated or partially unsaturated bicyclic heterocycle having 1 to 3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, a 5- to 6-membered monocyclic heteroaromatic ring having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and an 8- to 10-membered bicyclic heteroaromatic ring having 1 to 5 heteroatoms independently selected from nitrogen, oxygen, and sulfur; R 3 is hydrogen, halogen, -OR, or -OSiR3; R 3’ is hydrogen, halogen, -OR, or -OSiR3; or R 3 and R 3’ together to form =O or =S, R 4 and R 6 are independently hydrogen, —OR, —NR2, —NRCOR, —NRCO2R, —NRCONR2, —NRSO2R, —SR, —SO2NR2, or optionally substituted C 1~6 is aliphatic, R 5 and R 5’ are each hydrogen or together form =O or =NOR, X 1 and X 2 are each independently -CR2, -S-, or -S(O)-; X 1 and X 2 wherein at least one of the following is -CR2-; or a pharmaceutically acceptable salt thereof.

[0028] 2. Compounds and Definitions: The compounds of the present invention generally include those described herein and are further exemplified by the classes, subclasses, and species disclosed herein. As used herein, the following definitions shall apply unless otherwise indicated. For purposes of this invention, chemical elements are defined as defined in the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75 th In addition, general principles of organic chemistry are identified in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999 and "March's Advanced Organic Chemistry", 5 th Ed., Ed.: Smith, MB and March, J., John Wiley & Sons, New York: 2001, the entire contents of which are incorporated herein by reference.

[0029] As used herein, the terms "aliphatic" or "aliphatic group" mean a straight-chain (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is fully saturated or contains one or more units of unsaturation, or a monocyclic or bicyclic hydrocarbon that is fully saturated or contains one or more units of unsaturation, but is not aromatic, having a single point of attachment to the rest of the molecule (also referred to herein as "carbocyclic," "alicyclic," or "cycloalkyl"). Unless otherwise specified, an aliphatic group contains 1-6 aliphatic carbon atoms. In some embodiments, an aliphatic group contains 1-5 aliphatic carbon atoms. In other embodiments, an aliphatic group contains 1-4 aliphatic carbon atoms. In yet other embodiments, an aliphatic group contains 1-3 aliphatic carbon atoms, and in yet other embodiments, an aliphatic group contains 1-2 aliphatic carbon atoms. In some embodiments, "alicyclic" (or "carbocycle" or "cycloalkyl") refers to a monocyclic C3-C6 hydrocarbon that is fully saturated or contains one or more units of unsaturation, but is not aromatic, having a single point of attachment to the rest of the molecule. Suitable aliphatic groups include, but are not limited to, straight-chain or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl groups, and hybrids thereof, such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl, or (cycloalkyl)alkenyl.

[0030] The term "heteroatom" means one or more of oxygen, sulfur, nitrogen, phosphorus, or silicon (including any oxidized form of nitrogen, sulfur, phosphorus, or silicon, the quaternized form of any basic nitrogen, or a substitutable nitrogen of a heterocycle, e.g., N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or NR + (including as in N-substituted pyrrolidinyl).

[0031] As used herein, the term "unsaturated" means that a moiety has one or more units of unsaturation.

[0032] As used herein, "divalent C 1~8 (or C 1~6 The term "saturated or unsaturated, straight or branched hydrocarbon chain" refers to divalent alkylene, alkenylene, and alkynylene chains that are straight or branched as defined herein.

[0033] The term "alkylene" refers to a divalent alkyl group. An "alkylene chain" is a polymethylene group, i.e., -(CH2) n -, where n is a positive integer, preferably 1 to 6, 1 to 4, 1 to 3, 1 to 2, or 2 to 3. A substituted alkylene chain is a polymethylene group in which one or more methylene hydrogen atoms are replaced with a substituent. Suitable substituents include those described below for substituted aliphatic groups.

[0034] The term "alkenylene" refers to a divalent alkenyl group. A substituted alkenylene chain is a polymethylene group containing at least one double bond in which one or more hydrogen atoms are replaced with a substituent. Suitable substituents include those described below for substituted aliphatic groups.

[0035] The term "halogen" means F, Cl, Br, or I.

[0036] The term "aryl," used alone or as part of a larger moiety such as "aralkyl," "aralkoxy," or "aryloxyalkyl," refers to a monocyclic or bicyclic ring system having a total of 5 to 14 ring members, in which at least one ring of the system is aromatic and each ring of the system contains 3 to 7 ring members. The term "aryl" may be used interchangeably with the term "aryl ring." In certain embodiments of the present invention, "aryl" refers to an aromatic ring system, including, but not limited to, phenyl, biphenyl, naphthyl, anthracene, and the like, which may bear one or more substituents. Also included within the scope of the term "aryl," as used herein, are groups in which an aromatic ring is fused to one or more non-aromatic rings, such as indanyl, phthalimidyl, naphthymidyl, phenanthridinyl, or tetrahydronaphthyl.

[0037] The terms "heteroaryl" and "heteroari-," used alone or as part of a larger moiety, such as "heteroaralkyl" or "heteroalkoxy," refer to groups having 5 to 10 ring atoms, preferably 5, 6, or 9 ring atoms, with 6, 10, or 14 π electrons shared in the cyclic array, and having 1 to 5 heteroatoms in addition to carbon atoms. The term "heteroatom" refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur and any quaternized form of a basic nitrogen. Heteroaryl groups include, but are not limited to, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, and pteridinyl. As used herein, the terms "heteroaryl" and "heteroali-" also include groups in which a heteroaromatic ring is fused (the radical or point of attachment is on the heteroaromatic ring) with one or more aryl, alicyclic, or heterocyclyl rings. Non-limiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and pyrido[2,3-b]-1,4-oxazin-3(4H)-one. Heteroaryl groups can be monocyclic or bicyclic. The term "heteroaryl" may be used interchangeably with the terms "heteroaryl ring," "heteroaryl group," or "heteroaromatic," any of which terms include rings that are optionally substituted. The term "heteroaralkyl" refers to an alkyl group substituted by a heteroaryl, where the alkyl and heteroaryl portions independently are optionally substituted.

[0038] As used herein, the terms "heterocycle," "heterocyclyl," "heterocyclic radical," and "heterocyclic ring" are used interchangeably and refer to a stable 5- to 7-membered monocyclic moiety or a 7- to 10-membered bicyclic heterocyclic moiety that is either saturated or partially unsaturated and that, in addition to carbon atoms, has one or more (preferably 1-4) heteroatoms, as defined above. When used in reference to a ring atom of a heterocycle, the term "nitrogen" includes substituted nitrogen. By way of example, in a saturated or partially unsaturated ring having 0-3 heteroatoms selected from oxygen, sulfur, or nitrogen, the nitrogen can be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or + It can be NR (as in N-substituted pyrrolidinyl).

[0039] A heterocycle can be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure, and any of the ring atoms can be optionally substituted. Examples of such saturated or partially unsaturated heterocycle radicals include, but are not limited to, tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl. The terms "heterocycle," "heterocyclyl," "heterocyclyl ring," "heterocyclic group," "heterocyclic moiety," and "heterocyclic radical" are used interchangeably herein and include groups in which a heterocyclyl ring is fused to one or more aryl, heteroaryl, or alicyclic rings (e.g., indolinyl, 3H-indolyl, chromanyl, phenatridinyl, or tetrahydroquinolinyl). Heterocyclyl groups can be monocyclic or bicyclic (e.g., bridged bicyclic or spirocyclic). The term "heterocyclyl" refers to an alkyl group substituted by a heterocyclyl, wherein the alkyl and heterocyclyl portions independently are optionally substituted.

[0040] As used herein, the term "partially unsaturated" refers to a ring moiety that contains at least one double or triple bond. The term "partially unsaturated" is intended to encompass rings with multiple sites of unsaturation, but is not intended to include aryl or heteroaryl moieties as defined herein.

[0041] As described herein, compounds of the invention may contain "optionally substituted" moieties. Generally, the term "substituted" means that one or more hydrogens of the specified moiety are replaced with a suitable substituent. Unless otherwise specified, an "optionally substituted" group may have a suitable substituent at each substitutable position of the group, and if more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituents may be the same or different at all positions. Combinations of substituents envisioned by the present invention are preferably those that result in the formation of stable or chemically feasible compounds. As used herein, the term "stable" refers to compounds that are substantially unchanged when subjected to conditions that permit their production, detection, and, in certain embodiments, their recovery, purification, and use for one or more of the purposes disclosed herein.

[0042] Suitable monovalent substituents on a substitutable carbon atom of an "optionally substituted" group are independently halogen, -(CH) 0-4 R ○ , -(CH2) 0-4 OR ○ , -O(CH2) 0-4 R ○ , -O-(CH2) 0-4 C(O)OR ○ , -(CH2) 0-4 CH(OR ○ )2, -(CH2) 0-4 SR ○ , -(CH2) 0-4 Ph(R ○ (which may be substituted with -(CH2) 0-4 O(CH2) 0-1 Ph(R ○-CH=CHPh(R ○ (which may be substituted with -(CH2) 0-4 O(CH2) 0-1 -pyridyl (R ○ -(CH) 0-4 N(R ○ )2, -(CH2) 0-4 N(R ○ )C(O)R ○ , -N(R ○ )C(S)R ○ , -(CH2) 0-4 N(R ○ )C(O)NR ○ 2, -N(R ○ )C(S)NR ○ 2, -(CH2) 0-4 N(R ○ )C(O)OR ○ , -N(R ○ )N(R ○ )C(O)R ○ , -N(R ○ )N(R ○ )C(O)NR ○ 2, -N(R ○ )N(R ○ )C(O)OR ○ , -(CH2) 0-4 C(O)R ○ , -C(S)R ○ , -(CH2) 0-4 C(O)OR ○ , -(CH2) 0-4 C(O)SR ○ , -(CH2) 0-4 C(O)OSiR ○ 3. -(CH2) 0-4 OC(O)R ○ , -OC(O)(CH2) 0-4 SR-, SC(S)SR ○ , -(CH2) 0-4 SC(O)R ○ , -(CH2) 0-4 C(O)NR ○ 2. -C(S)NR ○ 2. -C(S)SR ○ , -SC(S)SR ○ , -(CH2) 0-4OC(O)NR ○ 2. -C(O)N(OR ○ )R ○ , -C(O)C(O)R ○ , -C(O)CHC(O)R ○ , -C(NOR ○ )R ○ , -(CH2) 0-4 SSR ○ , -(CH2) 0-4 S(O)2R ○ , -(CH2) 0-4 S(O)2OR ○ , -(CH2) 0-4 OS(O)2R ○ , -S(O)NR ○ 2, -(CH2) 0-4 S(O)R ○ , -N(R ○ )S(O)NR ○ 2, -N(R ○ )S(O)2R ○ , -N(OR ○ )R ○ , -C(NH)NR ○ 2. -P(O)2R ○ , -P(O)R ○ 2. -OP(O)R ○ 2. -OP(O)(OR ○ )2, SiR ○ 3, -(C 1~4 Linear or branched alkylene)ON(R ○ )2, or -(C 1~4 Linear or branched alkylene)C(O)ON(R ○ )2, where each R ○ may be substituted as defined below and independently represent hydrogen, C 1~6 Aliphatic, -CH2Ph, -O(CH2) 0-1 Ph, -CH2- (5- to 6-membered heteroaryl ring), or a 5- to 6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur; or, regardless of the above definitions, R ○two independent occurrences of together with their intervening atoms form a 3-12 membered saturated, partially unsaturated, or aryl monocyclic or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0043] R ○ The above suitable monovalent substituents (or R ○ The ring formed by two independent occurrences of -(CH2) together with their intervening atoms is independently a halogen, -(CH2) 0-2 R ● ,-(Halo R ● ), -(CH2) 0-2 OH, -(CH2) 0-2 OR ● , -(CH2) 0-2 CH(OR ● )2, -O(HaloR ● ), -CN, -N3, -(CH2) 0-2 C(O)R ● , -(CH2) 0-2 C(O)OH, -(CH2) 0-2 C(O)OR ● , -(CH2) 0-2 SR ● , -(CH2) 0-2 SH, -(CH2) 0-2 NH2, -(CH2) 0-2 NHR ● , -(CH2) 0-2 NR ● 2, -NO2, -SiR ● 3. -OSiR ● 3. -C(O)SR ● , -(C 1~4 Linear or branched alkylene)C(O)OR ● , or -SSR ● where each R ● is unsubstituted or, if preceded by "halo", is substituted with only one or more halogens, and independently represents C 1~4 Aliphatic, -CH2Ph, -O(CH2) 0-1 Ph, or a 5-6 membered saturated ring, partially unsaturated ring, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.○ Suitable divalent substituents on a saturated carbon atom of include ═O and ═S.

[0044] Suitable divalent substituents on a saturated carbon atom of an "optionally substituted" group include ═O, ═S, ═NNR * 2, =NNHC(O)R * , =NNHC(O)OR * , =NNHS(O)2R * , =NR * , =NOR * , -O(C(R * 2)) 2-3 O- or S(C(R * 2)) 2-3 Contains S-, R * Each independent occurrence of is hydrogen, C 1~6 The "optionally substituted" group is selected from an aliphatic (which may be substituted as defined below) or unsubstituted 5-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents attached to adjacent substitutable carbon atoms of the "optionally substituted" group include -O(CR * 2) 2-3 O-, wherein R * Each independent occurrence of is hydrogen, C 1~6 It is selected from an aliphatic (which may be substituted as defined below) or unsubstituted 5-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0045] R * Suitable substituents for the aliphatic group are independently halogen, -R ● ,-(Halo R ● ), -OH, -OR ● , -O(HaloR ● ), -CN, -C(O)OH, -C(O)OR ● , -NH2, -NHR ● , -NR ● 2, or -NO2, where each R ●is unsubstituted or, if preceded by "halo", is substituted with only one or more halogens, and independently represents C 1~4 Aliphatic, -CH2Ph, -O(CH2) 0-1 Ph, or a 5- to 6-membered saturated, partially unsaturated, or aryl ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0046] Suitable substituents on a substitutable nitrogen of an "optionally substituted" group include -R † , -NR † 2. -C(O)R † , -C(O)OR † , -C(O)C(O)R † , -C(O)CHC(O)R † , -S(O)2R † , -S(O)NR † 2. -C(S)NR † 2. -C(NH)NR † 2, or -N(R † )S(O)2R † wherein each R † are independently hydrogen, C 1~6 aliphatic (which may be substituted as defined below), unsubstituted -OPh, or an unsubstituted 5-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur; or, regardless of the above definitions, R † two independent occurrences of together with their intervening atoms form a 3-12 membered saturated, partially unsaturated, or aryl monocyclic or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0047] R † Suitable substituents for the aliphatic group are independently halogen, -R ● ,-(Halo R ● ), -OH, -OR ● , -O(HaloR ● ), -CN, -C(O)OH, -C(O)OR ● , -NH2, -NHR ● , -NR● 2, or -NO2, where each R ● is unsubstituted or, if preceded by "halo", is substituted with only one or more halogens, and independently represents C 1~4 Aliphatic, -CH2Ph, -O(CH2) 0-1 Ph, or a 5- to 6-membered saturated, partially unsaturated, or aryl ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0048] As used herein, the term "pharmaceutically acceptable salt" refers to a salt that is within the scope of sound medical common sense, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic reaction, etc., and commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, S.M. Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19 (incorporated herein by reference). Pharmaceutically acceptable salts of the compounds of the present invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are salts of amino groups formed with inorganic acids (e.g., hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid) or organic acids (e.g., acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid), or by using other methods used in the art, such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, and 2-hydroxy-ethanesulfonate. Examples of salts that can be used include benzoate, lactobionate, lactate, laurate, lauryl sulfate, malate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, palmitate, pamoate, pectinate, 3-phenylpropionate, phosphate, pivalate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, and valerate.

[0049] Salts derived from appropriate bases include alkali metal salts, alkaline earth metal salts, ammonium salts, and N + (C 1~4Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc. Further pharmaceutically acceptable salts include non-toxic ammonium, quaternary ammonium, and amine cations, formed where appropriate using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl sulfonates, and aryl sulfonates.

[0050] Unless otherwise indicated, compounds having the structures depicted herein are also meant to include any pharmaceutically acceptable salts, stereoisomers, tautomers, solvates, hydrates, and polymorphs thereof.

[0051] Unless otherwise stated, compounds having a structure depicted herein are also meant to include all isomeric (e.g., enantiomeric, diastereomeric, and geometric (or conformational) isomeric) forms of that structure, such as the R and S configurations for each asymmetric center, Z and E double bond isomers, and Z and E conformational isomers. Accordingly, single stereochemical isomers as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of the present compounds are within the scope of the invention. Unless otherwise stated, all tautomeric forms of the compounds of the invention are within the scope of the invention. Additionally, unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, the replacement of hydrogen by deuterium or tritium, or 13 C or 14 Compounds having this structure including the replacement of a carbon with a C-enriched carbon are within the scope of this invention. Such compounds are useful, for example, as analytical tools, probes in biological assays, or as therapeutic agents according to the present invention.

[0052] As used herein, the terms "measurable affinity" and "measurably inhibit" refer to a measurable change in mTORC1 activity between a sample containing a compound of the present invention or a composition thereof and mTORC1 and an equivalent sample containing mTORC1 in the absence of the compound or composition thereof.

[0053] 3. Description of Exemplary Embodiments: As noted above, in certain embodiments, the present invention provides a compound of formula I,

[0054] [ka] During the ceremony, X and X 3 are independently a covalent bond, -CR2-, -NR-, -NRCO-, -NRCO2-, -NRCONR-, -NRSO2-, -O-, -S-, or -SO2NR-; L 1 is a covalent bond or C 1~30 a divalent linear or branched saturated or unsaturated hydrocarbon chain in which 1 to 10 methylene units are independently and optionally replaced by -Cy1-, -O-, -S-, -SO2-, -C(O)-, -C(S)-, -CR2-, -CF2-, -P(O)(R)-, -SiR2-, -Si(OR)(R)-, or -NR-; each -Cy1- is independently an optionally substituted bivalent ring selected from phenylene, 4- to 7-membered saturated or partially unsaturated heterocyclylene having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and 5- to 6-membered heteroarylene having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; Each R is independently hydrogen or C 1~6 an optionally substituted group selected from aliphatic, 3- to 8-membered saturated or partially unsaturated monocyclic carbocycle, phenyl, 4- to 7-membered saturated or partially unsaturated heterocycle having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and 5- to 6-membered heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; or two R groups on the same atom, taken together with their intervening atoms, form a 4-7 membered saturated, partially unsaturated, or aryl ring having, in addition to the same atom to which they are attached, 0-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; L 2 optionally replaced by C 1~6 a divalent linear or branched saturated or unsaturated hydrocarbon chain, wherein one to two methylene units of said chain are independently and optionally replaced with -Cy1-; R 1 and R 2 are independently hydrogen, halogen, -OR, -CN, -NO2, -NR2, -NR(C 1~6 haloalkyl), -NRCO, -NRCO2R, -NRCONR2, -NRSO2R, -SR, -SO2NR2, or C 1~6 an optionally substituted group selected from an aliphatic, a 3- to 8-membered saturated or partially unsaturated monocyclic carbocycle, a phenyl, an 8- to 10-membered bicyclic aromatic carbocycle, a 4- to 8-membered saturated or partially unsaturated monocyclic heterocycle having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, and sulfur, a 7- to 10-membered saturated or partially unsaturated bicyclic heterocycle having 1 to 3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, a 5- to 6-membered monocyclic heteroaromatic ring having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and an 8- to 10-membered bicyclic heteroaromatic ring having 1 to 5 heteroatoms independently selected from nitrogen, oxygen, and sulfur; R 3 is hydrogen, halogen, -OR, or -OSiR3; R 3’ is hydrogen, halogen, -OR, or -OSiR3; or R 3 and R 3’ together to form =O or =S, R 4 and R 6 are independently hydrogen, —OR, —NR2, —NRCOR, —NRCO2R, —NRCONR2, —NRSO2R, —SR, —SO2NR2, or optionally substituted C 1~6 is aliphatic, R 5 and R 5’ are each hydrogen or together form =O or =NOR, X 1 and X 2 are each independently -CR2, -S-, or -S(O)-; X 1 and X 2 wherein at least one of the following is -CR2-; or a pharmaceutically acceptable salt thereof.

[0055] In certain embodiments, the present invention provides a compound of formula I':

[0056] [ka] During the ceremony, X and X 3 are independently a covalent bond, -CR2-, -NR-, -NRCO-, -NRCO2-, -NRCONR-, -NRSO2-, -O-, -S-, or -SO2NR-; L 1 is a covalent bond or C 1~30 a divalent linear or branched saturated or unsaturated hydrocarbon chain in which 1 to 10 methylene units are independently and optionally replaced by -Cy1-, -O-, -S-, -SO2-, -C(O)-, -C(S)-, -CR2-, -CF2-, -P(O)(R)-, -SiR2-, -Si(OR)(R)-, or -NR-; each -Cy1- is independently an optionally substituted bivalent ring selected from phenylene, 4- to 7-membered saturated or partially unsaturated heterocyclylene having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and 5- to 6-membered heteroarylene having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; Each R is independently hydrogen or C 1~6an optionally substituted group selected from aliphatic, 3- to 8-membered saturated or partially unsaturated monocyclic carbocycle, phenyl, 4- to 7-membered saturated or partially unsaturated heterocycle having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and 5- to 6-membered heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; or two R groups on the same atom, taken together with their intervening atoms, form a 4-7 membered saturated, partially unsaturated, or aryl ring having, in addition to the same atom to which they are attached, 0-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; L 2 optionally replaced by C 1~6 a divalent linear or branched saturated or unsaturated hydrocarbon chain, wherein one to two methylene units of said chain are independently and optionally replaced with -Cy1-; R 1 and R 2 are independently hydrogen, halogen, -OR, -CN, -(CR2) 1-4 NR2, -COR, -CONR2, -CONR(CR2) 1-4 NR2, -NO2, -NR2, -NR(C 1~6 haloalkyl), -NRCO, -NRCO2R, -NRCONR2, -NRSO2R, -SR, -SO2NR2, -P(O)R2, or C 1~6 an optionally substituted group selected from an aliphatic, a 3- to 8-membered saturated or partially unsaturated monocyclic carbocycle, a phenyl, an 8- to 10-membered bicyclic aromatic carbocycle, a 4- to 8-membered saturated or partially unsaturated monocyclic heterocycle having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, and sulfur, a 7- to 10-membered saturated or partially unsaturated bicyclic heterocycle having 1 to 3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, a 5- to 6-membered monocyclic heteroaromatic ring having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and an 8- to 10-membered bicyclic heteroaromatic ring having 1 to 5 heteroatoms independently selected from nitrogen, oxygen, and sulfur; R 3 is hydrogen, halogen, -OR, or -OSiR3; R 3’is hydrogen, halogen, -OR, or -OSiR3; or R 3 and R 3’ together to form =O or =S, R 4 and R 6 are independently hydrogen, —OR, —NR2, —NRCOR, —NRCO2R, —NRCONR2, —NRSO2R, —SR, —SO2NR2, or optionally substituted C 1~6 is aliphatic, R 5 and R 5’ are each hydrogen or together form =O or =NOR, X 1 and X 2 are each independently -CR2, -S-, or -S(O)-; X 1 and X 2 wherein at least one of the following is -CR2-; or a pharmaceutically acceptable salt thereof.

[0057] It will be understood that the term "rapamycin" and its structures recited throughout this specification are intended to encompass rapamycin and its analogs.

[0058] The analogs of rapamycin (ie, rapalogs) listed herein are for illustrative purposes only and are not intended to limit the invention.

[0059] X and X as defined above and described herein 3 are independently a covalent bond, -CR2-, -NR-, -NRCO-, -NRCO2-, -NRCONR-, -NRSO2-, -O-, -S-, or -SO2NR-.

[0060] In some embodiments, X is a covalent bond. In some embodiments, X is -CR2-. In some embodiments, X is -NR-. In some embodiments, X is -NRCO-. In some embodiments, X is -NRCO2-. In some embodiments, X is -NRCONR-. In some embodiments, X is -NRSO2-. In some embodiments, X is -O-. In some embodiments, X is -S-. In some embodiments, X is -SO2NR-.

[0061] In some embodiments, X 3 is a covalent bond. In some embodiments, X 3 is -CR-. In some embodiments, X 3 is -NR-. In some embodiments, X 3 is -NRCO-. In some embodiments, X 3 is -NRCO-. In some embodiments, X 3 is -NRCONR-. In some embodiments, X 3 is -NRSO2-. In some embodiments, X 3 is —O—. In some embodiments, X 3 is -S-. In some embodiments, X 3 is -SO2NR-.

[0062] In some embodiments where X is an unsymmetrical group, e.g., —NRCO—, —NRCO2—, —NRSO2—, or —SO2NR—, X is —NRCOL 1 -, -NRCO2L 1 -, -NRSO2L 1 -, and -SO2NRL 1 -L 1 Combine with.

[0063] X 3 In some embodiments, X is an asymmetric group, e.g., —NRCO—, —NRCO—, —NRSO—, or —SO—NR—. 3-NRCOR 2 , -NRCO2R 2 , -NRSO2R 2 , and -SO2NRR 2 as R 2 Combine with.

[0064] In some embodiments, X and X 3 is selected from those shown in the compounds of Table 1.

[0065] As defined above and described herein, L 1 is a covalent bond or C 1~30 A divalent straight or branched saturated or unsaturated hydrocarbon chain in which 1 to 10 methylene units are independently and optionally replaced with -Cy1-, -O-, -S-, -SO2-, -C(O)-, -C(S)-, -CR2-, -CF2-, -P(O)(R)-, -SiR2-, -Si(OR)(R)-, or -NR-.

[0066] In some embodiments, L 1 is a covalent bond. In some embodiments, L 1 is a covalent bond or C 1~30 A divalent straight or branched saturated or unsaturated hydrocarbon chain in which 1 to 10 methylene units are independently and optionally replaced with -Cy1-, -O-, -S-, -SO2-, -C(O)-, -C(S)-, -CR2-, -CF2-, -P(O)(R)-, -SiR2-, -Si(OR)(R)-, or -NR-.

[0067] In some embodiments, L 1 is -CH-. In some embodiments, L 1 is -CHCH-. In some embodiments, L 1 is —(CH)—. In some embodiments, L 1 is —(CH)—. In some embodiments, L 1 is —(CH)—. In some embodiments, L 1is —CH2CH2O—. In some embodiments, L 1 is -(CH2CH2O)2-. In some embodiments, L 1 is -(CH2CH2O)3-. In some embodiments, L 1 is -CH2CH2OCH2CH2-. In some embodiments, L 1 is —CH2CH2SO2CH2CH2O—. In some embodiments, L 1 is -CH2CH2OCH2CH2OCH2CH2-. In some embodiments, L 1 is -CH2CH2OCH2CH2-.

[0068] In some embodiments, L 1 is selected from those shown in the compounds of Table 1.

[0069] As defined above and described herein, each -Cy1- is independently an optionally substituted bivalent ring selected from phenylene, 4- to 7-membered saturated or partially unsaturated heterocyclylene having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and 5- to 6-membered heteroarylene having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0070] In some embodiments, -Cy1- is an optionally substituted bivalent ring selected from phenylene. In some embodiments, -Cy1- is an optionally substituted 4- to 7-membered saturated or partially unsaturated heterocyclylene having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, -Cy1- is an optionally substituted 5- to 6-membered heteroarylene having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0071] In some embodiments, -Cy1- is

[0072] [ka] is.

[0073] In some embodiments, -Cy1- is selected from those shown in the compounds of Table 1.

[0074] As defined above and described herein, each R is independently hydrogen or C 1~6 an optionally substituted group selected from an aliphatic, 3- to 8-membered saturated or partially unsaturated monocyclic carbocycle, phenyl, a 4- to 7-membered saturated or partially unsaturated heterocycle having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and a 5- to 6-membered heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; or two R groups on the same atom together with their intervening atoms form a 4- to 7-membered saturated, partially unsaturated, or aryl ring having, in addition to the same atom to which they are attached, 0-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0075] In some embodiments, R is hydrogen. In some embodiments, R is optionally substituted C 1~6 In some embodiments, R is C 1~6 In some embodiments, R is haloalkyl. In some embodiments, R is an optionally substituted 3-8 membered saturated or partially unsaturated monocyclic carbocycle. In some embodiments, R is an optionally substituted phenyl. In some embodiments, R is an optionally substituted 4-7 membered saturated or partially unsaturated heterocycle having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, R is an optionally substituted 5-6 membered heteroaryl ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, two R groups on the same atom, taken together with their intervening atoms, form an optionally substituted 4-7 membered saturated, partially unsaturated, or aryl ring having, in addition to the same atom to which they are attached, 0-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0076] In some embodiments, R is selected from those shown in the compounds of Table 1.

[0077] As defined above and described herein, L 2 is an optionally substituted C 1~6 A divalent straight or branched saturated or unsaturated hydrocarbon chain in which one to two methylene units are independently and optionally replaced with -Cyl-.

[0078] In some embodiments, L 2 is an optionally substituted C 1~6 A divalent straight or branched saturated or unsaturated hydrocarbon chain in which one to two methylene units are independently and optionally replaced with -Cyl-.

[0079] In some embodiments, L 2 teeth,

[0080] [ka] is.

[0081] In some embodiments, L 2 is selected from those shown in the compounds of Table 1.

[0082] R, as defined above and described herein, 1 and R 2 are independently hydrogen, halogen, -OR, -CN, -(CR2) 1-4 NR2, -COR, -CONR2, -CONR(CR2) 1-4 NR2, -NO2, -NR2, -NR(C 1~6 haloalkyl), -NRCO, -NRCO2R, -NRCONR2, -NRSO2R, -SR, -SO2NR2, -P(O)R2, or C 1~6an optionally substituted group selected from aliphatic, 3- to 8-membered saturated or partially unsaturated monocyclic carbocycle, phenyl, 8- to 10-membered bicyclic aromatic carbocycle, 4- to 8-membered saturated or partially unsaturated monocyclic heterocycle having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur, 7- to 10-membered saturated or partially unsaturated bicyclic heterocycle having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, 5- to 6-membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and 8- to 10-membered bicyclic heteroaromatic ring having 1-5 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0083] In some embodiments, R 1 is hydrogen. In some embodiments, R 1 is halogen. In some embodiments, R 1 is -OR. In some embodiments, R 1 is -CN. In some embodiments, R 1 is —NO. In some embodiments, R 1 is -NR2. In some embodiments, R 1 is -NR(C 1~6 haloalkyl). In some embodiments, R 1 is -NRCOR. In some embodiments, R 1 is -NRCO2R. In some embodiments, R 1 is -NRCONR2. In some embodiments, R 1 is -NRSO2R. In some embodiments, R 1 is -SR. In some embodiments, R 1 is -SO2NR2. In some embodiments, R 1 is an optionally substituted C 1~6 In some embodiments, R 1 is an optionally substituted 3-8 membered saturated or partially unsaturated monocyclic carbocyclic ring. In some embodiments, R 1is optionally substituted phenyl. In some embodiments, R 1 is an optionally substituted 8-10 membered bicyclic aromatic carbocycle. In some embodiments, R 1 is an optionally substituted 4-8 membered saturated or partially unsaturated monocyclic heterocycle having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, R 1 is an optionally substituted 7-10 membered saturated or partially unsaturated bicyclic heterocycle having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, R 1 is an optionally substituted 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, R 1 is an optionally substituted 8-10 membered bicyclic heteroaromatic ring having 1-5 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0084] In some embodiments, R 1 is methyl. In some embodiments, R 1 is -NH. In some embodiments, R 1 is -NHMe. In some embodiments, R 1 is -NMe. In some embodiments, R 1 is —CH 2 CF 3 . In some embodiments, R 1 is —SO—NH. In some embodiments, R 1 is -CONH. In some embodiments, R 1 is -CONMe. In some embodiments, R 1 is -OCONHMe. In some embodiments, R 1 is —COH. In some embodiments, R 1 teeth,

[0085] [ka] In some embodiments, R 1 teeth,

[0086] [ka] In some embodiments, R 1 teeth,

[0087] [ka] In some embodiments, R 1 teeth,

[0088] [ka] In some embodiments, R 1 teeth,

[0089] [ka] In some embodiments, R 1 teeth,

[0090] [ka] In some embodiments, R 1 teeth,

[0091] [ka] In some embodiments, R 1 teeth,

[0092] [ka] In some embodiments, R 1 teeth,

[0093] [ka] In some embodiments, R 1 teeth,

[0094] [ka] In some embodiments, R 1 teeth,

[0095] [ka] In some embodiments, R 1 teeth,

[0096] [ka] In some embodiments, R 1 teeth,

[0097] [ka] In some embodiments, R 1 teeth,

[0098] [ka] In some embodiments, R 1 teeth,

[0099] [ka] In some embodiments, R 1 teeth,

[0100] [ka] In some embodiments, R 1 teeth,

[0101] [ka] In some embodiments, R 1 teeth,

[0102] [ka] In some embodiments, R 1 teeth,

[0103] [ka] In some embodiments, R 1 teeth,

[0104] [ka] In some embodiments, R 1 teeth,

[0105] [ka] In some embodiments, R 1 teeth,

[0106] [ka] In some embodiments, R 1 teeth,

[0107] [ka] In some embodiments, R 1 teeth,

[0108] [ka] In some embodiments, R 1 teeth,

[0109] [ka] In some embodiments, R 1 teeth,

[0110] [ka] In some embodiments, R 1 teeth,

[0111] [ka] In some embodiments, R 1 teeth,

[0112] [ka] In some embodiments, R 1 teeth,

[0113] [ka] is.

[0114] In some embodiments, R 2 is hydrogen. In some embodiments, R 2 is halogen. In some embodiments, R 12 is -OR. In some embodiments, R 2 is -CN. In some embodiments, R 2 is -(CR2) 1-4 In some embodiments, R 2 is -COR. In some embodiments, R 2 is -CONR2. In some embodiments, R 2 -CONR(CR2) 1-4 In some embodiments, R 2 is —NO. In some embodiments, R 2 is -NR2. In some embodiments, R2 is -NR(C 1~6 haloalkyl). In some embodiments, R 2 is -NRCOR. In some embodiments, R 2 is -NRCO2R. In some embodiments, R 2 is -NRCONR2. In some embodiments, R 2 is -NRSO2R. In some embodiments, R 2 is -SR. In some embodiments, R 2 is -SO2NR2. In some embodiments, R 2 is -P(O)R. In some embodiments, R 2 is an optionally substituted C 1~6 In some embodiments, R 2 is an optionally substituted 3-8 membered saturated or partially unsaturated monocyclic carbocyclic ring. In some embodiments, R 2 is optionally substituted phenyl. In some embodiments, R 2 is an optionally substituted 8-10 membered bicyclic aromatic carbocycle. In some embodiments, R 2 is an optionally substituted 4-8 membered saturated or partially unsaturated monocyclic heterocycle having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, R 2 is an optionally substituted 7-10 membered saturated or partially unsaturated bicyclic heterocycle having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, R 2 is an optionally substituted 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, R 2 is an optionally substituted 8-10 membered bicyclic heteroaromatic ring having 1-5 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0115] In some embodiments, R 2 is methyl. In some embodiments, R2 is -CHF. In some embodiments, R 2 is -CF3. In some embodiments, R 2 teeth,

[0116] [ka] In some embodiments, R 2 teeth,

[0117] [ka] In some embodiments, R 2 teeth,

[0118] [ka] In some embodiments, R 2 teeth,

[0119] [ka] In some embodiments, R 2 teeth,

[0120] [ka] In some embodiments, R 2 teeth,

[0121] [ka] In some embodiments, R 2 teeth,

[0122] [ka] In some embodiments, R 2 teeth,

[0123] [ka] In some embodiments, R 2 teeth,

[0124] [ka] In some embodiments, R 2 teeth,

[0125] [ka] In some embodiments, R 2 teeth,

[0126] [ka] In some embodiments, R 2 teeth,

[0127] [ka] is.

[0128] In some embodiments, R 2 teeth,

[0129] [ka] In some embodiments, R 2 teeth,

[0130] [ka] In some embodiments, R 2 teeth,

[0131] [ka] In some embodiments, R 2 teeth,

[0132] [ka] In some embodiments, R 2 teeth,

[0133] [ka] is.

[0134] In some embodiments, R 2 teeth,

[0135] [ka] In some embodiments, R 2 teeth,

[0136] [ka] In some embodiments, R 2 teeth,

[0137] [ka] is.

[0138] In some embodiments, R 1 and R 2 is selected from those shown in the compounds of Table 1.

[0139] R, as defined above and described herein, 3 is hydrogen, halogen, —OR, or —OSiR3.

[0140] In some embodiments, R 3 is hydrogen. In some embodiments, R 3 is halogen. In some embodiments, R3 is -OR. In some embodiments, R 3 is -OSiR3.

[0141] In some embodiments, R 3 is -OMe.

[0142] R, as defined above and described herein, 3’ is hydrogen, halogen, —OR, or —OSiR3.

[0143] In some embodiments, R 3 is hydrogen. In some embodiments, R 3 is halogen. In some embodiments, R 3 is -OR. In some embodiments, R 3 is -OSiR3.

[0144] As defined above and described herein, in some embodiments, R 3 and R 3’ together to form =O or =S.

[0145] In some embodiments, R 3 and R 3’ together form =O. In some embodiments, R 3 and R 3’ together form =S.

[0146] In some embodiments, R 3 and R 3’ is selected from those shown in the compounds of Table 1.

[0147] R, as defined above and described herein, 4 and R 6 are independently hydrogen, -OR, -NR2, -NRCOR, -NRCO2R, -NRCONR2, -NRSO2R, -SR, -SO2NR2, or optionally substituted C 1~6 It is aliphatic.

[0148] In some embodiments, R 4 is hydrogen. In some embodiments, R 4 is -OR. In some embodiments, R 4 is -NR2. In some embodiments, R 4 is -NRCOR. In some embodiments, R 4 is -NRCO2R. In some embodiments, R 4 is -NRCONR2. In some embodiments, R 4 is -NRSO2R. In some embodiments, R 4 is -SR. In some embodiments, R 4 is -SO2NR2. In some embodiments, R 4 is an optionally substituted C 1~6 It is aliphatic.

[0149] In some embodiments, R 4 is —OH. In some embodiments, R 4 is -OMe.

[0150] In some embodiments, R 6 is hydrogen. In some embodiments, R 6 is -OR. In some embodiments, R 6 is -NR2. In some embodiments, R 6 is -NRCOR. In some embodiments, R 6 is -NRCO2R. In some embodiments, R 6 is -NRCONR2. In some embodiments, R 6 is -NRSO2R. In some embodiments, R 6 is -SR. In some embodiments, R 6 is -SO2NR2. In some embodiments, R 6 is an optionally substituted C 1~6 It is aliphatic.

[0151] In some embodiments, R 6 is -OMe.

[0152] In some embodiments, R 4 and R 6 is selected from those shown in the compounds of Table 1.

[0153] R, as defined above and described herein, 5 and R 5’ are each hydrogen or together form =O or =NOR.

[0154] In some embodiments, R 5 is hydrogen. In some embodiments, R 5’ is hydrogen. In some embodiments, R 5 and R 5’ together form =O. In some embodiments, R 5 and R 5’ together form =NOR.

[0155] In some embodiments, R 5 and R 5’ is selected from those shown in the compounds of Table 1.

[0156] X, as defined above and described herein 1 and X 2 are each independently -CR2-, -S-, or -S(O)-; X 1 and X 2 At least one of the groups is -CR2-.

[0157] In some embodiments, X 1 is -CR-. In some embodiments, X 1 is -S-. In some embodiments, X 1 is —S(O)—. In some embodiments, X 2 is -CR-. In some embodiments, X 2is -S-. In some embodiments, X 2 is -S(O)-.

[0158] In some embodiments, X 1 and X 2 is selected from those shown in the compounds of Table 1.

[0159] In certain embodiments, the present invention provides a compound of Formula I or Formula I', wherein X 1 is -CH2- and X 2 is -CH2- and L 2 but

[0160] [ka] thereby providing a compound of formula Ia-1,

[0161] [ka] During the ceremony, X is -CR2-, -NRCO-, -NRCO2-, -NRCONR-, -NRSO2-, or -SO2NR-; X 3 , R 1 , R 2 , R 3 , R 3’ , R 4 , R 5 , R 5’ , R 6 , and L 1 each of which, both individually and in combination, is as defined and described herein; or a pharmaceutically acceptable salt thereof.

[0162] In certain embodiments, the present invention provides a compound of Formula I or Formula I', wherein X 1 is -CH2- and X 2 is -CH2- and L 2 but

[0163] [ka] thereby providing a compound of formula Ia-2,

[0164] [ka] During the ceremony, X 3 is -CR2-, -NRCO-, -NRCO2-, -NRCONR-, -NRSO2-, or -SO2NR-, X, R 1 , R 2 , R 3 , R 3’ , R 4 , R 5 , R 5’ , R 6 , and L 1 each of which, both individually and in combination, is as defined and described herein; or a pharmaceutically acceptable salt thereof.

[0165] In certain embodiments, the present invention provides a compound of Formula I or Formula I', wherein X 1 is -CH2- and X 2 is -CH2- and L 2 but

[0166] [ka] thereby providing a compound of formula Ia-3,

[0167] [ka] During the ceremony, R 4 is -NR2, -NRRCOR, -NRCO2R, -NRCONR2, -NRSO2R, -SR, -SO2NR2, or an optionally substituted C 1~6 is aliphatic, X, X 3 , R 1 , R 2 , R 3 , R 3’ , R 5 , R 5’ , R 6 , and L 1 each of which, both individually and in combination, is as defined and described herein; or a pharmaceutically acceptable salt thereof.

[0168] In certain embodiments, the present invention provides a compound of Formula I or Formula I', wherein X 1 is -CH2- and X 2 is -CH2- and L 2 but

[0169] [ka] whereby a compound of formula Ia-4 is provided,

[0170] [ka] During the ceremony, R 6 is -NR2, -NRRCOR, -NRCO2R, -NRCONR2, -NRSO2R, -SR, -SO2NR2, or an optionally substituted C 1~6 is aliphatic, X, X 3 , R 1 , R 2 , R 3 , R 3’ , R 4 , R 5 , R 5’ , and L 1 each of which, both individually and in combination, is as defined and described herein; or a pharmaceutically acceptable salt thereof.

[0171] In certain embodiments, the present invention provides a compound of Formula I or Formula I', wherein X 1 is -CH2- and X 2 is -CH2- and L 2 but

[0172] [ka] and X 3 is a covalent bond and R 2 is hydrogen, thereby providing a compound of formula Ib-1,

[0173] [ka] During the ceremony, X, R 1 , R 3 , R 3’ , R 4 , R 5 , R 5’ , R 6 , and L 1 each of which, both individually and in combination, is as defined and described herein; or a pharmaceutically acceptable salt thereof.

[0174] In certain embodiments, the present invention provides a compound of Formula I or Formula I', wherein X 1 is -CH2- and X 2 is -CH2- and L 2 but

[0175] [ka] and R 3 is -OMe and R 3’ is hydrogen, thereby providing a compound of formula Ib-2,

[0176] [ka] During the ceremony, X, X 3 , R 1 , R 2 , R 4 , R 5 , R 5’ , R 6 , and L 1 each of which, both individually and in combination, is as defined and described herein; or a pharmaceutically acceptable salt thereof.

[0177] In certain embodiments, the present invention provides a compound of Formula I or Formula I', wherein X 1 is -CH2- and X 2 is -CH2- and L 2 but

[0178] [ka] and X 3 is -O- and R 2 but

[0179] [ka] thereby providing a compound of formula Ib-3,

[0180] [ka] During the ceremony, X, R 1 , R 3 , R 3’ , R 4 , R 5 , R 5’ , R 6 , and L 1 each of which, both individually and in combination, is as defined and described herein; or a pharmaceutically acceptable salt thereof.

[0181] Rapamycin is commercially available under the brand name Rapamune® (generic name: sirolimus) and is well known for its antiproliferative and immunosuppressive activities. Rapamycin is FDA-approved for the prevention of transplant rejection and for coating stents to prevent restenosis. Apart from its documented benefits, rapamycin is well known to be associated with many serious side effects. These include diabetes-like symptoms of decreased glucose tolerance and decreased insulin sensitivity. Additionally, rapamycin has been reported to increase patients' risk of cancer by activating the Akt signaling pathway (including activation of Akt and ERK).

[0182] As used herein, the phrase "rapamycin alone" is intended to compare the compounds of the present invention to rapamycin or an analog thereof (eg, everolimus as an alternative).

[0183] In some embodiments, provided compounds of Formula I or Formula I' are more effective than rapamycin alone.

[0184] In some embodiments, provided compounds of Formula Ia-1 through Formula Ia-5 are more effective than rapamycin alone.

[0185] In some embodiments, provided compounds of Formula Ib-1 through Formula Ib-3 are more effective than rapamycin alone.

[0186] In some embodiments, provided compounds of Formula I or Formula I', when administered to a patient, result in fewer and / or less severe side effects than when rapamycin is administered.

[0187] In some embodiments, provided compounds of Formula Ia-1 through Formula Ia-5, when administered to a patient, result in fewer and / or less severe side effects than when rapamycin is administered.

[0188] In some embodiments, provided compounds of Formula Ib-1 through Formula Ib-3, when administered to a patient, result in fewer and / or less severe side effects than when rapamycin is administered.

[0189] Exemplary compounds of the present invention are set forth in Table 1 below.

[0190] [Table 1-1]

[0191] [Table 1-2]

[0192] [Table 1-3]

[0193] [Table 1-4]

[0194] [Table 1-5]

[0195] [Table 1-6]

[0196] [Table 1-7]

[0197] [Table 1-8]

[0198] [Table 1-9]

[0199]

Table 1-10

[0200]

Table 1-11

[0201]

Table 1-12

[0202]

Table 1-13

[0203]

Table 1-14

[0204]

Table 1-15

[0205]

Table 1-16

[0206]

Table 1-17

[0207]

Table 1-18

[0208]

Table 1-19

[0209]

Table 1-20

[0210]

Table 1-21

[0211]

Table 1-22

[0212]

Table 1-23

[0213]

Table 1-24

[0214]

Table 1-25

[0215]

Table 1-26

[0216]

Table 1-27

[0217]

Table 1-28

[0218]

Table 1-29

[0219]

Table 1-30

[0220]

Table 1-31

[0221]

Table 1-32

[0222]

Table 1-33

[0223]

Table 1-34

[0224]

Table 1-35

[0225]

Table 1-36

[0226] In some embodiments, the present invention provides compounds set forth in Table 1 above, or pharmaceutically acceptable salts thereof. In some embodiments, the present invention provides compounds set forth in Table 1 above, or pharmaceutically acceptable salts, stereoisomers, tautomers, and polymorphs thereof. In some embodiments, the present invention provides compounds set forth in Table 1 above, comprising the replacement of one or more hydrogens with deuterium. It will also be understood that the present invention provides compounds set forth in Table 1 above, or pharmaceutically acceptable salts thereof, as a racemic mixture at the C7 position. It will further be understood that compounds set forth in Table 1 above, as a racemic mixture at the C7 hydroxyl position, can be separated into diastereomers by various methods (e.g., chiral chromatography).

[0227] In some embodiments, the present invention provides a compound of Formula I or Formula I', or a pharmaceutically acceptable salt thereof, wherein: L 2 but

[0228] [ka] and R 3 and R 3’ is hydrogen, R 5 and R 5’ together form =O, and X 1 and X 2 If both are -CH2-, -XL 1 -R 1 , -X 3 -R 2 , R 4 , and R 6 are present in combinations other than those presented in each row of Table 1A below.

[0229] [Table 2-1]

[0230]

Table 2-2

[0231]

Table 2-3

[0232]

Table 2-4

[0233]

Table 2-5

[0234]

Table 2-6

[0235]

Table 2-7

[0236]

Table 2-8

[0237]

Table 2-9

[0238]

Table 2-10

[0239]

Table 2-11

[0240] In some embodiments, the invention provides compounds of Formula I or Formula I' that are not one or more of the compounds of Table 1A. In some embodiments, the invention provides compounds of Formula I or Formula I', excluding compounds of Table 1A, or pharmaceutically acceptable salts, stereoisomers, tautomers, and polymorphs thereof, in which one or more hydrogens are replaced by deuterium.

[0241] In some embodiments, the present invention provides a compound of Formula I or Formula I', or a pharmaceutically acceptable salt, stereoisomer, tautomer, or polymorph thereof, wherein: L 2 but

[0242] [ka] and R 3 and R 3’ is hydrogen, R 5 and R 5’ together form =O, and X 1 and X 2 If both are -CH2-, -XL 1 -R 1 , -X 3 -R 2 , R 4 , and R 6 are present in combinations other than those presented in each row of Table 1A below.

[0243] 4. Use, Formulation, and Administration Pharmaceutically acceptable compositions According to another embodiment, the present invention provides a composition, comprising a compound of the present invention or a pharmaceutically acceptable derivative thereof and a pharmaceutically acceptable carrier, adjuvant, or vehicle. The amount of the compound in the composition of the present invention is such that it is effective to measurably inhibit mTORC1 in a biological sample or a patient. In certain embodiments, the amount of the compound in the composition of the present invention is such that it is effective to measurably inhibit mTORC1 in a biological sample or a patient. In certain embodiments, the composition of the present invention is formulated for administration to a subject (e.g., a patient) in need of such a composition. In some embodiments, the composition of the present invention is formulated for oral administration to a patient.

[0244] The term "patient", as used herein, means an animal, preferably a mammal, and most preferably a human.

[0245] The term "pharmaceutically acceptable carrier, adjuvant, or vehicle" refers to a non-toxic carrier, adjuvant, or vehicle that does not destroy the pharmacological activity of the compound with which it is formulated. Pharmaceutically acceptable carriers, adjuvants, or vehicles that can be used in the compositions of the present invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins (e.g., human serum albumin), buffer substances (e.g., phosphoric acid), glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes (e.g., protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol, and wool fat.

[0246] The compositions of the present invention may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, bucally, intravaginally, or via an implanted reservoir. As used herein, the term "parenteral" includes subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional, and intracranial injection or infusion techniques. Preferably, the compositions are administered orally, intraperitoneally, or intravenously. Sterile injectable forms of the compositions of the present invention may be aqueous or oily suspensions. These suspensions can be formulated using suitable dispersing or wetting agents and suspending agents according to techniques known in the art. Sterile injectable preparations may also be sterile injectable solutions or suspensions in non-toxic parenterally acceptable diluents or solvents, such as solutions in 1,3-butanediol. Among acceptable vehicles and solvents that may be used are water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally used as solvents or suspending media.

[0247] For this purpose, any bland fixed oil can be used, including synthetic monoglycerides or diglycerides. Fatty acids such as oleic acid and its glyceride derivatives are useful for preparing injectables, as are natural pharmaceutically acceptable oils, such as olive oil or castor oil, especially their polyoxyethylated versions. These oil solutions or suspensions can also contain long-chain alcohol diluents or dispersants, such as carboxymethylcellulose, or similar dispersants commonly used in the formulation of pharmaceutically acceptable dosage forms, including emulsions and suspensions. Other commonly used surfactants, such as Tween, Span, and other emulsifiers or bioavailability enhancers (which are commonly used in the production of pharmaceutically acceptable solid, liquid, or other dosage forms), can also be used for formulation purposes.

[0248] The pharmaceutically acceptable composition of the present invention can be orally administered in any orally acceptable dosage form, including but not limited to capsules, tablets, pills, aqueous suspensions or aqueous solutions.For tablets for oral use, commonly used carriers include lactose and corn starch.Lubricants such as magnesium stearate are also typically added.For oral administration in capsule form, useful diluents include lactose and dried corn starch.When aqueous suspensions are required for oral use, active ingredients are combined with emulsifiers and suspending agents.If desired, certain sweeteners, flavorings or colorings can also be added.

[0249] Alternatively, the pharmaceutically acceptable compositions of the present invention can be administered in the form of suppositories for rectal administration. These can be prepared by mixing the drug with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature, and therefore melts in the rectum to release the drug. Such materials include cocoa butter, beeswax, and polyethylene glycol.

[0250] The pharmaceutically acceptable compositions of this invention may also be administered topically, particularly when the target of treatment includes areas or organs readily accessible by topical application, including diseases of the eye, the skin, or the lower intestinal tract. Suitable topical formulations are readily prepared for each of these areas or organs.

[0251] Topical application for the lower intestinal tract can be effected in a rectal suppository formulation (see above) or in a suitable enema formulation. Topical-transdermal patches can also be used.

[0252] For topical application, the provided pharmaceutically acceptable composition can be formulated into a suitable ointment containing the active ingredient suspended or dissolved in one or more carriers.Carriers for topical administration of the compounds of the present invention include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compounds, emulsifying wax, and water.Alternatively, the provided pharmaceutically acceptable composition can be formulated into a suitable lotion or cream containing the active ingredient suspended or dissolved in one or more pharmaceutically acceptable carriers.Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl ester wax, cetearyl alcohol, 2 octyldodecanol, benzyl alcohol, and water.

[0253] For ophthalmic use, the provided pharmaceutically acceptable compositions can be formulated as a micronized suspension in isotonic, pH-adjusted, sterile saline, or, preferably, as a solution in isotonic, pH-adjusted, sterile saline, either with or without a preservative, such as benzylalkonium chloride. Alternatively, for ophthalmic use, the pharmaceutically acceptable compositions can be formulated into an ointment, such as petrolatum.

[0254] The pharmaceutically acceptable compositions of this invention may also be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption enhancers to enhance bioavailability, fluorocarbons, and / or other conventional solubilizing or dispersing agents.

[0255] Most preferably, the pharmaceutically acceptable compositions of the present invention are formulated for oral administration. Such formulations can be administered with or without food. In some embodiments, the pharmaceutically acceptable compositions of the present invention are administered without food. In other embodiments, the pharmaceutically acceptable compositions of the present invention are administered with food.

[0256] The amount of the compounds of the present invention that can be combined with the carrier materials to produce a composition in a single dosage form will vary depending on the host being treated and the particular mode of administration. Preferably, the compositions provided should be formulated so that a dosage of 0.01 to 100 mg / kg body weight / day of the inhibitor can be administered to a patient receiving these compositions.

[0257] It is understood that the specific dosage and treatment regimen for any particular patient will depend on a variety of factors, including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, and the judgment of the treating physician, as well as the severity of the particular disease being treated. The amount of a compound of the invention in a composition will also depend on the particular compound in the composition.

[0258] Uses of the Compounds and Pharmaceutically Acceptable Compositions As used herein, the terms "treatment," "treat," and "treating," as described herein, refer to reversing, alleviating, slowing, or inhibiting the progression of a disease or disorder, or one or more symptoms thereof. In some embodiments, treatment may be administered after one or more symptoms have developed. In other embodiments, treatment may be administered in the absence of symptoms. For example, treatment may be administered to a susceptible individual prior to the onset of symptoms (e.g., in light of a history of the symptoms and / or genetic or other susceptibility factors). Treatment may also be continued after symptoms have resolved, e.g., to prevent or delay their recurrence.

[0259] The provided compounds are inhibitors of mTORC1 and are therefore useful for treating one or more disorders associated with the activity of mTORC1. Accordingly, in certain embodiments, the present invention provides a method for treating an mTORC1-mediated disorder, comprising administering to a patient in need thereof a compound of the present invention, or a pharmaceutically acceptable composition thereof.

[0260] As used herein, the term "mTORC1-mediated" disorder, disease, and / or condition refers to any disease or other deleterious condition in which mTORC1 is known to play a role. Accordingly, another embodiment of the present invention relates to treating or lessening the severity of one or more diseases in which mTORC1 is known to play a role. In certain embodiments, the mTORC1-mediated disorder, disease, and / or condition is selected from those described by Matt Kaeberlin, Scientifica, vol. 2013, Article ID 849186.

[0261] The methods described herein include methods for treating cancer in a subject. As used in this context, "treating" means alleviating or improving at least one symptom or clinical parameter of cancer. For example, treatment may result in a decrease in tumor size or growth rate. Treatment does not necessarily result in a 100% cure or remission of cancer in all subjects.

[0262] As used herein, the term "cancer" refers to cells capable of autonomous growth (i.e., an abnormal state or condition characterized by rapidly proliferating cell proliferation). The term "cancer" is meant to include all types of cancerous growths or oncogenic processes, metastatic tissues, or malignantly transformed cells, tissues, or organs, regardless of histopathological type or stage of invasiveness. The term "tumor," as used herein, refers to a population of cancerous cells, e.g., cancer cells.

[0263] Cancers that can be treated or diagnosed using the methods described herein include malignancies of various organ systems (e.g., those affecting the lung, breast, thyroid, lymphatic system, gastrointestinal, and genitourinary tract), as well as adenocarcinomas (which include malignancies such as most colon cancers, renal cell carcinoma, prostate cancer, and / or testicular tumors, non-small cell lung cancer, small intestine cancer, and esophageal cancer).

[0264] In some embodiments, the methods described herein are used to treat or diagnose carcinoma in a subject. The term "carcinoma" is art-recognized and refers to a malignant tumor of epithelial or endocrine tissue, including respiratory system carcinoma, gastrointestinal system carcinoma, genitourinary system carcinoma, testicular cancer, breast cancer, prostate carcinoma, endocrine system carcinoma, and melanoma. In some embodiments, the cancer is renal carcinoma and melanoma. Exemplary carcinomas include those forming from tissue of the cervix, lung, prostate, breast, head and neck, colon, and ovary. The term also includes carcinosarcomas, which include, for example, malignant tumors composed of carcinomatous and sarcomatous tissue. "Adenocarcinoma" refers to a carcinoma derived from glandular tissue or in which the tumor cells form recognizable glandular structures.

[0265] The term "sarcoma" is art-recognized and refers to malignant tumors of mesenchymal derivation.

[0266] In some embodiments, the cancer treated by the methods described herein is a cancer that has an increased level of mTORC1 or increased expression or activity of mTORC1 compared to normal tissue or other cancers of the same tissue, and these cancers can be identified using methods known in the art and described herein. In some embodiments, the method includes obtaining a sample containing cancer cells, determining mTORC1 activity in the sample, and administering a treatment described herein (e.g., a provided mTORC1 inhibitor). In some embodiments, the cancer is a cancer shown herein to have an increased level of mTORC1 activity.

[0267] In some embodiments, the present invention provides methods for treating one or more disorders, diseases, and / or conditions, including but not limited to, cell proliferative disorders.

[0268] Cell proliferative disorders The present invention features methods and compositions for the diagnosis and prognosis of cell proliferative disorders (e.g., cancer) and the treatment of these disorders by inhibiting mTORC1 activity. Cell proliferative disorders described herein include, for example, cancer, obesity, and proliferation-dependent diseases. Such disorders can be diagnosed using methods known in the art.

[0269] cancer Cancers include, but are not limited to, leukemias (e.g., acute leukemia, acute lymphocytic leukemia, acute myeloid leukemia, acute myeloblastic leukemia, acute promyelocytic leukemia, acute myelomonocytic leukemia, acute monocytic leukemia, acute erythroleukemia, chronic leukemia, chronic myelogenous leukemia, chronic lymphocytic leukemia), polycythemia vera, lymphomas (e.g., Hodgkin's disease, non-Hodgkin's disease), Waldenstrom's hypergammaglobulinemia, multiple myeloma, heavy chain disease, and solid tumors, such as sarcomas and carcinomas (e.g., fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endothelial sarcoma, lymphangiosarcoma, lymphangioendothelial sarcoma, synovial sarcoma, and sarcoma). tumor, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon cancer, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinoma, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatocellular carcinoma, cholangiocarcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilms' tumor, cervical cancer, uterine cancer, testicular cancer, lung cancer, small cell lung cancer, bladder cancer, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, schwannoma, meningioma, melanoma, neuroblastoma, and retinoblastoma). In some embodiments, the cancer is melanoma or breast cancer.

[0270] fibrotic disorders Idiopathic pulmonary fibrosis (IPF). The PI3K pathway is activated in fibrotic lesions, the primary lesion of IPF. The mTOR kinase inhibitor GSK2126458 reduces PI3K pathway signaling and functional responses in IPF-derived lung fibroblasts, and in a patient model of IPF, mTOR inhibition reduces collagen expression. In a bleomycin model of pulmonary fibrosis, rapamycin treatment is antifibrotic, and rapamycin also reduces alpha-smooth muscle actin and fibronectin expression by fibroblasts in vitro.

[0271] In some embodiments, methods of inhibiting mTORC1 activity are used to treat idiopathic pulmonary fibrosis (IPF) (see Mercer, PF et al., Thorax., 71(8):701-11 (2016); Patel, AS, et al., PLoS One, 7(7):e41394 (2012)). Accordingly, in some embodiments, the present invention provides methods of treating idiopathic pulmonary fibrosis (IPF) in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0272] Renal fibrosis. mTORC1 is activated in myofibroblasts, the major pathogenic cell type in renal fibrosis. Inhibition of mTOR with rapamycin in a mouse model of renal fibrosis (UUO) attenuates the expression of markers of fibrosis and tubulointerstitial injury.

[0273] In some embodiments, methods of inhibiting mTORC1 activity are used to treat renal fibrosis (see Jiang, L., et al., J Am Soc Nephrol, 24(7):1114-26(2013); Wu, MJ et al., Kidney International, 69(11):2029-36(2006); Chen, G. et al., PLoS One, 7(3):e33626(2012); Liu, C F et al., Clin Invest Med, 37(34):E142-53(2014)). Accordingly, in some embodiments, the present invention provides methods of treating renal fibrosis in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0274] In some embodiments, methods of inhibiting mTORC1 activity are used to treat scleroderma (see Mitra, A., et al., J Invest Dermatol. 135(11):2873-6 (2015)). Accordingly, in some embodiments, the present invention provides a method of treating scleroderma in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0275] In some embodiments, methods of inhibiting mTORC1 activity are used to treat hypertrophic scars and keloid disease (see Syed, F., et al., Am J Pathol. 181(5):1642-58 (2012)). Accordingly, in some embodiments, the present invention provides a method of treating hypertrophic scars and keloid disease in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0276] In some embodiments, methods of inhibiting mTORC1 activity are used to treat cardiac fibrosis (see Yano, T., et al., J Mol Cell Cardiol. 91:6-9 (2016)). Accordingly, in some embodiments, the present invention provides methods of treating cardiac fibrosis in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0277] Other proliferative disorders Other proliferative diseases include, for example, obesity, benign prostatic hyperplasia, psoriasis, abnormal keratinization, lymphoproliferative disorders (e.g., disorders in which there is abnormal proliferation of cells of the lymphatic system), rheumatoid arthritis, arteriosclerosis, restenosis, and diabetic retinopathy. Proliferative diseases include those described in U.S. Patent Nos. 5,639,600 and 7,087,648 (incorporated herein by reference).

[0278] Other disorders Other disorders include lysosomal storage diseases (including, but not limited to, Pompe disease, Gaucher disease, mucopolysaccharidoses, multiple sulfatase deficiency), neurodegenerative diseases (e.g., Parkinson's disease, Alzheimer's disease, Huntington's disease, alpha-1 antitrypsin deficiency, and spinal-bulbar muscular atrophy).

[0279] In some embodiments, methods of inhibiting mTORC1 activity are used to treat asthma (see Hua, W., et al., Respirology, 20(7):1055-65 (2015)). Accordingly, in some embodiments, the present invention provides methods of treating asthma in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0280] In some embodiments, methods of inhibiting mTORC1 activity are used to treat lysosomal storage diseases (see Sardiello, M., Annals of the New York Academy of Sciences, 1371(1):3-14 (2016); Awad, O., et al., Hum Mol Genet. 24(20):5775-88 (2015); Spampanato, C., et al., EMBO Mol Med., 5(5):691-706 (2013); Medina, DL, et al., Dev Cell., 21(3):421-30 (2011)). Accordingly, in some embodiments, the present invention provides methods of treating lysosomal storage diseases in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0281] In some embodiments, methods of inhibiting mTORC1 activity are used to treat Parkinson's disease (see Decressac, M., et al., Proc Natl Acad Sci U S A., 110(19):E1817-26 (2013)). Accordingly, in some embodiments, the present invention provides methods of treating Parkinson's disease in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0282] In some embodiments, methods of inhibiting mTORC1 activity are used to treat Alzheimer's disease (see Polito, VA, et al., EMBO Mol Med. 6(9):1142-60 (2014)). Accordingly, in some embodiments, the present invention provides methods of treating Alzheimer's disease in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0283] In some embodiments, methods of inhibiting mTORC1 activity are used to treat Huntington's disease (see Tsunemi, T., et al., Sci Transl Med., 4(142):142ra97 (2012)). Accordingly, in some embodiments, the present invention provides methods of treating Huntington's disease in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0284] In some embodiments, methods of inhibiting mTORC1 activity are used to treat alpha 1 antitrypsin deficiency (see Pastore, N. et al., EMBO Mol Med., 5(3):397-412 (2013)). Accordingly, in some embodiments, the present invention provides methods of treating alpha 1 antitrypsin deficiency in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0285] In some embodiments, methods of inhibiting mTORC1 activity are used to treat spinal-bulbar muscular atrophy (see Cortes, CJ, et al., Nat Neurosci., 17(9):1180-9 (2014)). Accordingly, in some embodiments, the present invention provides a method of treating spinal-bulbar muscular atrophy in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0286] In some embodiments, the method of inhibiting mTORC1 activity is used to treat Fragile X syndrome (FXS), amyotrophic lateral sclerosis (ALS), epilepsy, focal cortical dysplasia (FCD), hemimegalencephaly (HME), familial focal epilepsy with variable foci (FFEV), temporal lobe epilepsy (TLE), seizures, neurodegenerative diseases, Down syndrome, Rett syndrome (RTS), or diseases associated with activation or overactivation of mTOR signaling in the brain.

[0287] In some embodiments, the present invention provides a method of treating Fragile X Syndrome (FXS) in a patient in need thereof, comprising administering a compound of the present invention or a pharmaceutically acceptable salt thereof.

[0288] In some embodiments, the present invention provides a method of treating amyotrophic lateral sclerosis (ALS) in a patient in need thereof, comprising administering a compound of the present invention or a pharmaceutically acceptable salt thereof.

[0289] In some embodiments, the present invention provides a method of treating epilepsy in a patient in need thereof, comprising administering a compound of the present invention or a pharmaceutically acceptable salt thereof.

[0290] In some embodiments, the present invention provides a method of treating focal cortical dysplasia (FCD) in a patient in need thereof, comprising administering a compound of the present invention or a pharmaceutically acceptable salt thereof.

[0291] In some embodiments, the present invention provides a method of treating hemimegalencephaly (HME) in a patient in need thereof, comprising administering a compound of the present invention or a pharmaceutically acceptable salt thereof.

[0292] In some embodiments, the present invention provides a method of treating familial focal epilepsy with variable foci (FFEV) in a patient in need thereof, comprising administering a compound of the present invention or a pharmaceutically acceptable salt thereof.

[0293] In some embodiments, the present invention provides a method of treating temporal lobe epilepsy (TLE) in a patient in need thereof, comprising administering a compound of the present invention or a pharmaceutically acceptable salt thereof.

[0294] In some embodiments, the present invention provides a method of treating seizures in a patient in need thereof, comprising administering a compound of the present invention or a pharmaceutically acceptable salt thereof.

[0295] In some embodiments, the present invention provides a method of treating a neurodegenerative disease in a patient in need thereof, comprising administering a compound of the present invention or a pharmaceutically acceptable salt thereof.

[0296] In some embodiments, the present invention provides a method of treating Down's syndrome in a patient in need thereof, comprising administering a compound of the present invention or a pharmaceutically acceptable salt thereof.

[0297] In some embodiments, the present invention provides a method of treating Rett Syndrome (RTS) in a patient in need thereof, comprising administering a compound of the present invention or a pharmaceutically acceptable salt thereof.

[0298] In some embodiments, the present invention provides a method of treating a disease associated with activation or overactivation of mTOR signaling in the brain in a patient in need thereof, comprising administering a compound of the present invention or a pharmaceutically acceptable salt thereof.

[0299] In some embodiments, the compounds of the present invention bind to FKBP12 to form a complex. In some embodiments, the complex between the compounds of the present invention and FKBP12 interacts with the FK506-rapamycin binding domain of mTOR.

[0300] In some embodiments, the compounds of the invention bind to FKBP12 and disrupt the protein-protein interaction between FRAP and FKBP12. 1 The group interacts with both FRAP and FKBP12.

[0301] The present invention provides compounds that are inhibitors of mTORC1 activity and have been shown to selectively inhibit mTORC1 over mTORC2 as measured by pS6K inhibition (a measure of mTORC1 activity) and pAKT activation (a measure of mTORC2 activity). In some embodiments, provided compounds selectively inhibit mTORC1 over mTORC2. In some embodiments, provided compounds do not measurably inhibit mTORC2. In some embodiments, provided compounds exhibit an IC of pAKT activation. 50is greater than 10 μM. In some embodiments, provided compounds inhibit mTORC1 with greater than 10-fold selectivity over mTORC2. In some embodiments, provided compounds inhibit mTORC1 with greater than 20-fold selectivity over mTORC2. In some embodiments, provided compounds inhibit mTORC1 with greater than 50-fold selectivity over mTORC2. In some embodiments, provided compounds inhibit mTORC1 with greater than 100-fold selectivity over mTORC2. In some embodiments, provided compounds inhibit mTORC1 with greater than 150-fold selectivity over mTORC2. In some embodiments, provided compounds inhibit mTORC1 with greater than 200-fold selectivity over mTORC2. In some embodiments, provided compounds inhibit mTORC1 with greater than 500-fold selectivity over mTORC2. In some embodiments, provided compounds inhibit mTORC1 with greater than 1000-fold selectivity over mTORC2.

[0302] In some embodiments, provided compounds selectively inhibit mTORC1 over mTORC2 after chronic treatment or exposure. In some embodiments, provided compounds selectively inhibit mTORC1 over mTORC2 about 24 hours after treatment or exposure. In some embodiments, provided compounds selectively inhibit mTORC1 over mTORC2 about 36 hours after treatment or exposure. In some embodiments, provided compounds selectively inhibit mTORC1 over mTORC2 about 48 hours after treatment or exposure. In some embodiments, provided compounds selectively inhibit mTORC1 over mTORC2 about 72 hours after treatment or exposure. In some embodiments, provided compounds selectively inhibit mTORC1 over mTORC2 about 96 hours after treatment or exposure. In some embodiments, provided compounds selectively inhibit mTORC1 over mTORC2 about 120 hours after treatment or exposure. In some embodiments, provided compounds selectively inhibit mTORC1 over mTORC2 about 144 hours after treatment or exposure. In some embodiments, provided compounds selectively inhibit mTORC1 over mTORC2 after about 1 week of treatment or exposure. In some embodiments, provided compounds selectively inhibit mTORC1 over mTORC2 after more than about 1 week of treatment or exposure.

[0303] In some embodiments, provided compounds are less immunosuppressive than existing rapalogs. In some embodiments, provided compounds are less immunosuppressive than rapamycin. In some embodiments, provided compounds are less immunosuppressive than everolimus. In some embodiments, provided compounds are less immunosuppressive than temsirolimus. In some embodiments, provided compounds are less immunosuppressive than ridaforolimus. In some embodiments, provided compounds are less immunosuppressive than umirolimus.

[0304] In some embodiments, provided compounds suppress interferon gamma (IFN-γ) production to a lesser extent than rapalogs. In some embodiments, provided compounds suppress IFN-γ production to a lesser extent than rapamycin. In some embodiments, provided compounds suppress IFN-γ production to a lesser extent than everolimus. In some embodiments, provided compounds suppress IFN-γ production to a lesser extent than temsirolimus. In some embodiments, provided compounds suppress IFN-γ production to a lesser extent than ridaforolimus. In some embodiments, provided compounds suppress IFN-γ production to a lesser extent than umirolimus.

[0305] In some embodiments, provided compounds reduce the expression of fibrosis biomarkers in injured tissue. In some embodiments, provided compounds reduce the expression of collagen I (COL1A2) in injured tissue. In some embodiments, provided compounds reduce the expression of collagen III (COL3A1) in injured tissue. In some embodiments, provided compounds reduce the expression of fibronectin (FN1) in injured tissue.

[0306] In some embodiments, provided compounds reduce the tendency of immune cells to infiltrate damaged tissue, hi some embodiments, provided compounds reduce the tendency of macrophage cells to infiltrate damaged tissue.

[0307] In some embodiments, provided compounds induce lower glucose tolerance than rapalogs. In some embodiments, provided compounds induce lower glucose tolerance than rapamycin. In some embodiments, provided compounds induce lower glucose tolerance than everolimus. In some embodiments, provided compounds induce lower glucose tolerance than temsirolimus. In some embodiments, provided compounds induce lower glucose tolerance than ridaforolimus. In some embodiments, provided compounds induce lower glucose tolerance than umirolimus. In some embodiments, provided compounds do not induce significantly higher glucose tolerance than placebo or vehicle alone.

[0308] Thus, in some embodiments, the present invention provides methods of treating disorders associated with mTORC1, comprising administering to a patient a compound that inhibits mTORC1, where the compound does not inhibit mTORC2. Such compounds may be used in indications for which rapamycin and rapalogs have shown benefit in either animal models or human disease settings. Such indications include: Treatment of metabolic diseases (obesity and insulin resistance in type 2 diabetes). Inhibition of the mTORC1 pathway leads to lifespan extension in yeast, flies, and mice, and calorie restriction improves lifespan and insulin sensitivity. The underlying mechanism is proposed to function through modulation of mTORC1 activation. Rapamycin-induced insulin resistance has been shown to be mediated by inhibition of mTORC2, and selective mTORC1 inhibitors are predicted to improve insulin sensitivity and glucose homeostasis.

[0309] In some embodiments, methods of inhibiting mTORC1 activity are used to treat metabolic diseases (obesity and insulin resistance in type 2 diabetes) (see Yu, Z., et al., J Gerontol A Biol Sci Med Sci, 70(4), 410-20 (2015); Fok, WC, et al., Aging Cell 13(2):311-9 (2014); Shum, M., et al., Diabetologia, 59(3):592-603 (2016); Lamming, DW, et al., Science 335(6076):1638-43 (2012)). Accordingly, in some embodiments, the present invention provides methods of treating metabolic diseases (obesity and insulin resistance in type 2 diabetes) in a patient in need of such treatment, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0310] Neurofibromatosis. Neurofibromatosis type 1 (NF1) is caused by mutations in the NF1 gene. Its protein product, neurofibromin, functions as a tumor suppressor, ultimately leading to constitutive upregulation of mTOR. mTOR inhibitors have been shown to reduce tumor size and induce antiproliferative effects in NF1-associated plexiform neurofibromas.

[0311] In some embodiments, methods of inhibiting mTORC1 activity are used to treat neurofibromatosis (see Franz, DN, et al., Curr Neurol Neurosci Rep., 12(3):294-301 (2012); Varin, J., et al., Oncotarget., 7:35753-67 (2016)). Accordingly, in some embodiments, the present invention provides methods of treating neurofibromatosis in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0312] A model of cardiomyopathy and skeletal muscle dystrophy, Emery-Dreifuss muscular dystrophy (LMNA) - / - Mutations in LMNA result in several human diseases, including limb-girdle muscular dystrophy (LGMD1B), Emery-Dreifuss muscular dystrophy (EDMD2 / 3), dilated cardiomyopathy (DCM), and conduction system disease (CMD1A), lipodystrophy, Charcot-Marie-Tooth disease, and Hutchinson-Gilford progeria syndrome (HGPS). - / - Mice have elevated mTORC1 activity and Lmna - / - Short-term treatment with rapamycin in mice reduces mTORC1 signaling, improves cardiac and skeletal muscle function, and increases survival by approximately 50%.

[0313] In some embodiments, methods of inhibiting mTORC1 activity are used to treat cardiomyopathy and skeletal muscular dystrophy (see Ramos, F., et al., Sci Transl Med., 4(144):144ral03 (2012); Bonne, G. & Quijano-Roy, S., Handb Clin Neurol., 113:1367-76 (2013)). Accordingly, in some embodiments, the present invention provides methods of treating cardiomyopathy and skeletal muscular dystrophy in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0314] Leigh syndrome. Ndufs4 knockout (KO) mice are used as a model of Leigh syndrome and exhibit mTORC1 hyperactivation and metabolic disorders. Treatment of Ndufs4 KO mice with rapamycin extends lifespan and ameliorates the metabolic and neurological disorders associated with the disease.

[0315] In some embodiments, methods of inhibiting mTORC1 activity are used to treat Leigh syndrome (see Johnson, SC, et al., Science, 342(6165):1524-8(2013)). Accordingly, in some embodiments, the present invention provides a method of treating Leigh syndrome in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0316] Oncology. Inhibition of mTOR by rapalogs has been shown to have antitumor activity in mouse cancer models and cancer patients. Examples of susceptible cancer types include, but are not limited to, hepatocellular carcinoma, breast cancer, mantle cell lymphoma, lung cancer, tuberous sclerosis complex, and lymphangioleiomyomatosis.

[0317] In some embodiments, methods of inhibiting mTORC1 activity are used to treat cancer and oncological disorders (see Ilagan, E. & manning, BD, Trends Cancer, 2(5):241-51 (2016)). Accordingly, in some embodiments, the present invention provides methods of treating cancer and oncological disorders in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0318] Non-alcoholic steatohepatitis (NASH) The present invention provides an inhibitor that induces autophagy, which removes degraded cytoplasmic proteins. NASH disease is characterized by lipid deposition, inflammation, and fibrosis in the liver. Inhibition of the mTORC1 pathway induces autophagy and downregulates SREBP-1, reducing lipid biosynthesis and decreasing lipid storage.

[0319] In some embodiments, methods of inhibiting mTORC1 activity are used to treat non-alcoholic steatohepatitis (NASH) (see Puri, P. & Chandra, A., J Clin Exp Hepatol, 4(1):51-9 (2014)). Accordingly, in some embodiments, the present invention provides methods of treating non-alcoholic steatohepatitis (NASH) in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0320] Tuberous sclerosis (TSC) and lymphangioleiomyomatosis (LAM). Dysregulation of mTOR is critical to the pathogenesis of the genetic disorder tuberous sclerosis complex (TSC) and the related lung disease lymphangioleiomyomatosis (LAM). Both diseases are caused by mutations in TSC1 or TSC2, leading to inappropriate activation of signaling downstream of mTORC1. Patients with TSC develop nonmalignant tumors in many organs, including the brain, while patients with LAM (predominantly women) accumulate abnormal muscle-like cells in specific organs or tissues, particularly the lungs, lymph nodes, and kidneys. The rapalogs everolimus and sirolimus are currently approved by the U.S. FDA for the treatment of TSC and LAM, respectively.

[0321] In some embodiments, methods of inhibiting mTORC1 activity are used to treat tuberous sclerosis and lymphangioleiomyomatosis (see Wander, SA, et al., J. Clin. Invest., 121(4):1231-41 (2011); Taveira-DaSilva, AM & Moss, J., J. Clin Epidemiol., 7:249-57 (2015)). Accordingly, in some embodiments, the present invention provides methods of treating tuberous sclerosis and lymphangioleiomyomatosis in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0322] Senescence and age-related diseases. Rapamycin inhibits the mammalian TORC1 complex, which regulates translation, and extends lifespan in various species, including mice. Rapamycin has been shown to inhibit the proinflammatory phenotype of senescent cells. As senescent cells accumulate with age, the senescence-associated secretory phenotype (SASP) may disrupt tissues and contribute to age-related pathologies, including cancer. Inhibition of mTOR suppressed the secretion of proinflammatory cytokines by senescent cells. Rapamycin reduced cytokine levels, including IL6, and suppressed the translation of the membrane-bound cytokine IL1A. Reduction of IL1A reduces the transcriptional activity of NF-κB and regulates SASP. Therefore, mTORC1 inhibitors may ameliorate age-related pathologies, including cancer, in later life by suppressing senescence-associated inflammation.

[0323] In some embodiments, methods of inhibiting mTORC1 activity are used to treat aging and age-related diseases (see Laberge, RM, et al., Nature Cell Biology, 17(8):1049-61 (2015); Nacarelli, T., et al., Free Radic Biol Med., 95:133-54 (2016)). Accordingly, in some embodiments, the present invention provides methods of treating aging and age-related diseases in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0324] Diabetic Nephropathy and Kidney-Related Complications in Type 1 and Type 2 Diabetes. Diabetic nephropathy is a kidney complication in type 1 and type 2 diabetes, affecting up to approximately 40% of diabetic patients. High levels of glucose in the kidneys overwork them to filter blood, resulting in kidney damage. Research suggests that the mTOR pathway is highly activated in patients with diabetic neuropathy and may be involved in the pathological changes and renal dysfunction caused by chronic high glucose. Furthermore, mTOR inhibition can attenuate hyperinsulinemia.

[0325] In some embodiments, methods of inhibiting mTORC1 activity are used to treat diabetic nephropathy or kidney-related complications in type 1 and type 2 diabetes (see Mori, H., et al., Biochem. Res. Commun. 384(4):471-5 (2009)). Accordingly, in some embodiments, the present invention provides methods of treating diabetic nephropathy or kidney-related complications in type 1 and type 2 diabetes in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0326] Polycystic kidney disease (PKD) is characterized by the development and accumulation of destructive kidney cysts, which ultimately lead to kidney failure. PKD can be autosomal dominant (ADPKD) or recessive (ARPKD). Dysfunction of the mTOR signaling pathway has been observed in ADPKD and ARPKD. Therefore, normalizing the mTORC1 pathway can ameliorate cyst development and disease progression.

[0327] In some embodiments, methods of inhibiting mTORC1 activity are used to treat PKD (see Torres, VE, et al., Clin. J. Am. Soc. Nephrol. 5(7):1312-29 (2010)). Accordingly, in some embodiments, the present invention provides methods of treating PKD in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof. In some embodiments, the PKD is autosomal dominant. In some embodiments, the PKD is autosomal recessive.

[0328] Focal segmental glomerulosclerosis (FSGS) and other diseases associated with nephrosclerosis. FSGS is the most common primary glomerular disorder leading to end-stage renal disease (ESRD) in the United States. As the disease progresses, there is a discrepancy between the surface area of ​​podocytes in Bowman's capsule and the glomerular basement membrane they cover. Studies have shown that podocyte size is regulated by mTOR and that mTOR activation contributes to disease progression. Furthermore, constitutive activation of mTORC1 has been shown to induce FSGS-like lesions in mouse knockdown experiments. Therefore, inhibiting mTORC1 may ameliorate FSGS or other diseases associated with nephrosclerosis by normalizing or increasing autophagy activity.

[0329] In some embodiments, methods of inhibiting mTORC1 activity are used to treat FSGS or other diseases associated with nephrosclerosis (see Zschiedrich, S. et al., J. Am. Soc. Nephrol. 28(7):2144-57 (2017)). Accordingly, in some embodiments, the present invention provides methods of treating FSGS or other diseases associated with nephrosclerosis in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0330] Age-related macular degeneration. Age-related macular degeneration (AMD) is a leading cause of blindness characterized by the death of photoreceptors in the macula. Possible mechanisms of AMD progression include oxidative stress, which leads to protein deposition and organelle dysfunction, leading to hypertrophy, dedifferentiation, and eventual atrophy of the retinal pigment epithelium. mTOR is involved in the dedifferentiation of the retinal pigment epithelium. Therefore, inhibition of mTORC1 can ameliorate AMD by blocking hypertrophy and dedifferentiation.

[0331] In some embodiments, methods of inhibiting mTORC1 activity are used to treat age-related macular degeneration (see Kolosova, N.G., et al., Am. J. Path. 181(2):472-7 (2012) and Zhen, C. & Vollrath, D., Aging 3(4):346-47 (2011)). Accordingly, in some embodiments, the present invention provides methods of treating age-related macular degeneration in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0332] Diabetic macular edema (DME) is the leading cause of blindness in diabetic patients, affecting approximately 35% of diabetic patients. Research suggests that the pathogenesis of DME is an inflammatory disease involving various cytokines and chemokines. Chronic inflammatory and oxidative stress may contribute to the progression of DME. Therefore, inhibition of mTORC1 may improve the symptoms and progression of DME by reducing the inflammatory response.

[0333] In some embodiments, methods of inhibiting mTORC1 activity are used to treat DME (see Okamoto, T., et al., PLOS ONE, (11)(1):e0146517, https: / / doi.org / 10.1371 / journal.pone.0146517(2016)). Accordingly, in some embodiments, the present invention provides methods of treating DME in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0334] Diabetic retinopathy. Diabetic retinopathy (DR) is a common eye disease that accounts for approximately 5% of blindness in adults and is associated with chronic hyperglycemia and defects in the insulin signaling pathway. DR patients suffer from persistent damage to retinal blood vessels and neurons due to inflammation, reactive oxygen species, and endoplasmic reticulum stress caused by chronic hyperglycemia. Importantly, rapamycin has been shown to be beneficial in blocking the action of insulin-induced hypoxia-inducible factor-1 (HIF-1) and retinal cell senescence, inducing autophagy, promoting apoptosis of new blood vessels, and preventing angiogenesis. Therefore, inhibition of mTORC1 may improve the symptoms and progression of DR by reducing inflammation and inhibiting pathogenic signaling pathways.

[0335] In some embodiments, methods of inhibiting mTORC1 activity are used to treat DR (see Di Rosa, M., et al., Curr. Neuropharmacol. 14(8):810-25 (2016)). Accordingly, in some embodiments, the present invention provides methods of treating DR in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0336] Glaucoma. Glaucoma is a common optic nerve disorder associated with aging and elevated intraocular pressure and is a leading cause of irreversible blindness. Research suggests that mTOR-dependent dysregulation of autophagy may contribute to disease progression. Therefore, inhibiting mTORC1 can slow the progression or ameliorate glaucoma by normalizing or increasing autophagy.

[0337] In some embodiments, methods of inhibiting mTORC1 activity are used to treat glaucoma (see Porter, K., et al., Biochim. Biophys. Acta. 1852(3):379-85 (2014)). Accordingly, in some embodiments, the present invention provides methods of treating glaucoma in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0338] Restoration of immune function. Inhibition of mTORC1 inhibits CD4 + and CD8 + It has been shown that mTORC1 reduces the expression of the programmed death-1 (PD-1) receptor and promotes T cell signaling in T lymphocytes. Thus, inhibition of mTORC1 can restore immune function by improving adaptive immune responses.

[0339] In some embodiments, methods of inhibiting mTORC1 activity are used to restore immune function (see Mannick, JB, et al., Sci. Trans. Med. 6(268):ppra179(2014)). Accordingly, in some embodiments, the present invention provides methods of restoring immune function in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0340] Treatment of respiratory and / or urinary tract infections. Inhibition of mTORC1 may reduce infections by upregulating antiviral gene expression and responses. Therefore, inhibition of mTORC1 can enhance the ability of a patient's immune system to defend against respiratory tract infections and / or urinary tract infections.

[0341] In some embodiments, methods of inhibiting mTORC1 activity are used to treat respiratory tract infections and / or urinary tract infections. (See Mannick, JB, et al., Sci. Trans. Med. 10(449):eaaq1564(2018)). Accordingly, in some embodiments, the present invention provides methods of restoring immune function in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0342] Heart failure. mTORC1 activity is essential for cardiac hypertrophy in response to stress, but can also cause cardiac derangement as a result of cardiac remodeling after infarction. Inhibition of mTORC1 reduces cardiac remodeling and heart failure in response to pressure overload. Therefore, inhibition of mTORC1 may reduce heart failure in patients with myocardial damage.

[0343] In some embodiments, methods of inhibiting mTORC1 activity are used to treat heart failure (see Sciarretta, S. et al., Circ. Res. 122(3):489-505 (2018)). Accordingly, in some embodiments, the present invention provides methods of treating heart failure in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0344] Osteoarthritis. Osteoarthritis (OA) is a chronic degenerative disease that leads to cartilage loss and arthritis. mTOR plays an important role in collagen homeostasis and turnover and cartilage remodeling. Therefore, inhibiting mTORC1 can slow or improve the progression of osteoarthritis symptoms by normalizing cartilage turnover.

[0345] In some embodiments, methods of inhibiting mTORC1 activity are used to treat osteoarthritis (see Pal, B., et al., Drugs R&D, 15(1):27-36 (2017)). Accordingly, in some embodiments, the present invention provides methods of treating osteoarthritis in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0346] Pulmonary arterial hypertension (PAH) is a progressive, fatal disease associated with elevated pulmonary vascular resistance. Smooth muscle cell proliferation and migration in the pulmonary artery contribute to the progression of arterial wall thickening and exacerbate vasoconstriction. Therefore, mTORC1 inhibition can alleviate PAH by reducing vascular remodeling.

[0347] In some embodiments, methods of inhibiting mTORC1 activity are used to treat PAH (see Ma, X., et al., Interact. Cardiovasc. Thorac. Surg. 25(2):206-11 (2017)). Accordingly, in some embodiments, the present invention provides methods of treating PAH in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0348] Chronic obstructive pulmonary disease. Reduced autophagy leads to the accumulation of proteins and other cellular substances that promote cellular senescence in patients with chronic obstructive pulmonary disease (COPD). Therefore, inhibiting mTORC1 can slow the progression or improve COPD symptoms by normalizing or increasing autophagy.

[0349] In some embodiments, methods of inhibiting mTORC1 activity are used to treat COPD (see Fujii, S., et al., Oncoimmunology 1(5):630-41(2012)). Accordingly, in some embodiments, the present invention provides methods of treating COPD in a patient in need thereof, comprising administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0350] Additional therapeutic indications in which mTORC inhibition may be beneficial include cardiovascular disease (acute coronary syndrome), coronary artery occlusion due to eluting stents, polycystic kidney disease, and cyst formation. These include: kidney diseases associated with cyst formation or cystogenesis, neurofibromatosis, epilepsy associated with TSC1 / TSC2 mutations, polycystic liver disease, pachyonychia congenita, fragile X syndrome, Friedreich's ataxia, Peutz-Jeghers syndrome, eye diseases (including neovascular age-related macular degeneration), uveitis, diabetic macular edema, fibroblast proliferation (including pulmonary fibrosis), renal failure / fibrosis, metabolic syndrome, diseases of the immune system (including immunosenescence), lupus nephritis, chronic immune thrombocytopenia, multiple sclerosis, cancer (including lymphoma), tumors associated with TSC1 / 2 mutations, angiomyolipoma associated with TSC1 / 2 mutations, breast cancer, hepatocellular carcinoma, leukemia, glioma, adenoid cystic carcinoma, aging, autism, and vascular rheumatoid arthritis.

[0351] In some embodiments, methods of inhibiting mTORC1 activity are used to treat cardiovascular disease (acute coronary syndrome), coronary artery occlusion due to an eluting stent, polycystic kidney disease, neurofibromatosis, epilepsy associated with TSC1 / TSC2 mutations, polycystic liver disease, pachyonychia congenita, fragile X syndrome, Friedreich's ataxia, Peutz-Jeghers syndrome, eye diseases (including age-related macular degeneration with neovascularization), uveitis, diabetic macular edema, fibroblast proliferation (including pulmonary fibrosis), renal failure / fibrosis, metabolic syndrome, diseases of the immune system (including immunosenescence), lupus nephritis, chronic immune thrombocytopenia, multiple sclerosis, cancer (including lymphoma), tumors associated with TSC1 / 2 mutations, angiomyolipoma associated with TSC1 / 2 mutations, breast cancer, hepatocellular carcinoma, leukemia, glioma, adenoid cystic carcinoma, aging, autism, and rheumatoid arthritis.

[0352] Thus, in some embodiments, the present invention provides a method for treating cardiovascular disease (acute coronary syndrome), coronary artery occlusion due to eluting stents, polycystic kidney disease, neurofibromatosis, epilepsy associated with TSC1 / TSC2 mutations, polycystic liver disease, pachyonychia congenita, fragile X syndrome, Friedreich's ataxia, Peutz-Jeghers syndrome, eye diseases (including neovascular age-related macular degeneration), uveitis, diabetic macular edema, fibroblast proliferation (including pulmonary fibrosis), renal failure / fibrosis, metabolic disease. Also provided are methods of treating TSC1 / 2 mutation-associated syndromes, diseases of the immune system (including immunosenescence), lupus nephritis, chronic immune thrombocytopenia, multiple sclerosis, cancer (including lymphoma), tumors associated with TSC1 / 2 mutations, angiomyolipoma associated with TSC1 / 2 mutations, breast cancer, hepatocellular carcinoma, leukemia, glioma, adenoid cystic carcinoma, aging, autism, and vascular rheumatoid arthritis in a patient in need of such treatment, comprising the step of administering to the patient a provided compound or a pharmaceutically acceptable salt thereof.

[0353] The pharmaceutically acceptable compositions of the present invention may be administered to humans and other animals orally, rectally, parenterally, intracisternally, intravaginally, intraperitoneally, topically (by powder, ointment, or drops), buccally, or via oral or nasal spray, etc., depending on the severity of the infection being treated. In certain embodiments, the compounds of the present invention may be administered orally or parenterally, once or twice or more times daily, at dosage levels of about 0.01 mg / kg to about 50 mg / kg, preferably about 1 mg / kg to about 25 mg / kg of subject body weight per day, to achieve the desired therapeutic effect.

[0354] Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs.In addition to the active compound, liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilizers and emulsifiers (e.g., ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide), oils (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitan, and mixtures thereof.In addition to inert diluents, oral compositions may also contain adjuvants (e.g., wetting agents, emulsifiers, and suspending agents), and suspending agents, sweeteners, flavoring agents, and fragrances.

[0355] Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions, can be formulated according to known techniques using suitable dispersing or wetting agents and suspending agents. Sterile injectable preparations may also be sterile injectable solutions, suspensions, or emulsions in non-toxic parenterally acceptable diluents or solvents, for example, solutions in 1,3-butanediol. Acceptable vehicles and solvents that can be used include water, Ringer's solution, USP standard, and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally used as solvents or suspending media. For this purpose, any non-irritating fixed oil can be used, including synthetic monoglycerides or diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables.

[0356] Injectable preparations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium before use.

[0357] In order to prolong the effect of a compound of the present invention, it is often desirable to slow the absorption of the compound from subcutaneous or intramuscular injection. This can be accomplished by using a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the compound then depends on its dissolution rate, which may in turn depend on crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered compound form can be achieved by dissolving or suspending the compound in an oil vehicle. Injectable depot forms are made by forming microencapsule matrices of the compound in biodegradable polymers such as polylactide-polyglycolide. The release rate of the compound can be controlled depending on the ratio of compound to polymer and the nature of the particular polymer used. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the compound in liposomes or microemulsions that are compatible with body tissues.

[0358] Compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing a compound of the present invention with a suitable non-irritating excipient or carrier, such as cocoa butter, polyethylene glycol, or a suppository wax which is solid at ambient temperature but liquid at body temperature and therefore melts in the rectum or vaginal cavity to release the active compound.

[0359] Oral solid dosage forms include capsules, tablets, pills, powders, and granules.In such solid dosage forms, the active compound is mixed with at least one inert pharmaceutically acceptable excipient or carrier, such as sodium citrate or dicalcium phosphate, and / or a) fillers or extenders (such as starch, lactose, sucrose, glucose, mannitol, and silicic acid), b) binders (such as carboxymethylcellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and acacia), c) humectants (such as glycerol), d) disintegrants (such as agar, calcium carbonate, , potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate), e) solution retarders (e.g., paraffin), f) absorption accelerators (e.g., quaternary ammonium compounds), g) wetting agents (e.g., acetyl alcohol and glycerol monostearate), h) absorbents (e.g., kaolin and bentonite clay), and i) lubricants (e.g., talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate), and mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may also contain buffering agents.

[0360] Solid compositions of a similar type may also be used as fillers in soft and hard-filled gelatin capsules using excipients such as lactose or milk sugar, as well as high molecular weight polyethylene glycols. Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical formulation art. These may optionally contain opacifying agents and may also be of a composition that releases the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. Solid compositions of a similar type may also be used as fillers in soft and hard-filled gelatin capsules using excipients such as lactose or milk sugar, as well as high molecular weight polyethylene glycols.

[0361] The active compound may also be in microencapsulated form with one or more excipients, as described above. Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells, such as enteric coatings, release-controlling coatings, and other coatings well known in the pharmaceutical formulation art. In such solid dosage forms, the active compound may be mixed with at least one inert diluent, such as sucrose, lactose, or starch. Such dosage forms may also contain, as is customary, additional substances other than inert diluents, such as tableting lubricants and other tableting aids, such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets, and pills, the dosage forms may also contain buffering agents. These may optionally contain opacifying agents, and may also be of a composition that releases the active ingredient only, or preferentially, in a certain part of the intestinal tract, optionally in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes.

[0362] Dosage forms for topical or transdermal administration of the compounds of the present invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, or patches. The active ingredient is mixed under sterile conditions with a pharmaceutically acceptable carrier and any necessary preservatives or buffers, as needed. Ophthalmic formulations, ear drops, and eye drops are also contemplated within the scope of the present invention. In addition, the present invention contemplates the use of transdermal patches, which have the added advantage of providing controlled delivery of the compound into the body. Such dosage forms can be made by dissolving or dispensing the compound in a suitable medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate can be controlled by providing a rate-controlling membrane or by dispersing the compound in a polymer matrix or gel.

[0363] The term "biological sample," as used herein, includes, but is not limited to, cell cultures or extracts thereof; biopsies obtained from mammals or extracts thereof; and blood, saliva, urine, feces, semen, tears, or other bodily fluids or extracts thereof.

[0364] In another embodiment, the present invention provides a method for treating an mTORC1-mediated disorder in a patient in need thereof, comprising administering to said patient a compound according to the present invention or a pharmaceutically acceptable composition thereof. These disorders are described in detail herein.

[0365] Depending on the particular condition, or disease, being treated, additional therapeutic agents, which are normally administered to treat that condition, may also be present in the compositions of this invention. As used herein, additional therapeutic agents that are normally administered to treat a particular disease, or condition, are known as "appropriate for the disease, or condition, being treated."

[0366] The compounds of the present invention may also be advantageously used in combination with other antiproliferative compounds, including, but not limited to, aromatase inhibitors, antiestrogens, topoisomerase I inhibitors, topoisomerase II inhibitors, microtubule active compounds, alkylating compounds, histone deacetylase inhibitors, compounds that induce cell differentiation processes, cyclooxygenase inhibitors, MMP inhibitors, mTOR inhibitors, antitumor antimetabolites, platinum compounds, compounds that target / reduce protein or lipid kinase activity, and further antiangiogenic compounds, compounds that target, reduce or inhibit the activity of protein or lipid phosphatases, gonadorelin agonists, antiandrogens, and the like. anti-cancer drugs, methionine aminopeptidase inhibitors, matrix metalloproteinase inhibitors, bisphosphonates, biological response modifiers, anti-proliferative antibodies, heparanase inhibitors, inhibitors of Ras oncogenic isoforms, telomerase inhibitors, proteasome inhibitors, compounds used in the treatment of hematological malignancies, compounds that target, reduce, or inhibit the activity of Flt-3, Hsp90 inhibitors (e.g., 17-AAG (17-allylaminogeldanamycin, NSC330507), 17-DMAG (17-dimethylaminoethylamino-17-demethoxy-geldanamycin, NSC707545), Conforma Therapeutics), IPI-504, CNF1010, CNF2024, CNF1010), temozolomide (Temodal®), kinesin spindle protein inhibitors (such as SB715992 or SB743921 from GlaxoSmithKline, or pentamidine / chlorpromazine from CombinatoRx), MEK inhibitors (such as ARRY142886 from Array BioPharma, AZD6244 from AstraZeneca, PD181461 from Pfizer, and leucovorin). The term "aromatase inhibitor" as used herein relates to a compound that inhibits estrogen production (e.g., converting the substrates androstenedione and testosterone to estrone and estradiol, respectively).The term includes, but is not limited to, steroids, particularly atamestane, exemestane, and formestane, and nonsteroidal agents, particularly aminoglutethimide, rogletimide, pyridoglutethimide, trilostane, testolactone, ketoconazole, vorozole, fadrozole, anastrozole, and letrozole. Exemestane is commercially available under the trade name Aromasin™. Formestane is commercially available under the trade name Lentaron™. Fadrozole is commercially available under the trade name Afema™. Anastrozole is commercially available under the trade name Arimidex™. Letrozole is commercially available under the trade name Femara™ or Femar™. Aminoglutethimide is commercially available under the trade name Orimeten™. Combinations of the present invention that include a chemotherapeutic agent that is an aromatase inhibitor are particularly useful in the treatment of hormone receptor-positive tumors, such as breast tumors.

[0367] The term "antiestrogen" as used herein refers to a compound that antagonizes the effects of estrogen at the estrogen receptor level. This term includes, but is not limited to, tamoxifen, fulvestrant, raloxifene, and raloxifene hydrochloride. Tamoxifen is commercially available under the trade name Nolvadex™. Raloxifene hydrochloride is commercially available under the trade name Evista™. Fulvestrant can be administered under the trade name Faslodex™. The combination of the present invention, which includes a chemotherapeutic agent that is an antiestrogen, is particularly useful for treating estrogen receptor-positive tumors, such as breast tumors.

[0368] The term "antiandrogen" as used herein refers to any substance capable of inhibiting the biological effects of androgen hormones, including, but not limited to, bicalutamide (Casodex™). The term "gonadorelin agonist" as used herein includes, but is not limited to, abarelix, goserelin, and goserelin acetate. Goserelin can be administered under the trade name Zoladex™.

[0369] The term "topoisomerase I inhibitor" as used herein includes, but is not limited to, topotecan, dimatecan, irinotecan, camptothecin, and its analogs 9-nitrocamptothecin and PNU-166148, a polymeric camptothecin conjugate. Irinotecan can be administered in a form such as that marketed under the trade name Camptosar™. Topotecan is marketed under the trade name Hycamptin™.

[0370] The term "topoisomerase II inhibitors" as used herein includes, but is not limited to, anthracyclines (e.g., doxorubicin (including liposomal formulations such as Caelyx™), daunorubicin, epirubicin, idarubicin, and nemorubicin), anthraquinones (mitoxantrone) and losoxantrone), and podophyllotoxins (etoposide and teniposide). Etoposide is commercially available under the trade name Etopophos™. Teniposide is commercially available under the trade name VM26-Bristol. Doxorubicin is commercially available under the trade name Acriblastin™ or Adriamycin™. Epirubicin is commercially available under the trade name Farmorubicin™. Idarubicin is commercially available under the trade name Zavedos™. Mitoxantrone is commercially available under the trade name Novantron.

[0371] The term "microtubule active agent" refers to microtubule-stabilizing compounds, microtubule-destabilizing compounds, and microtubule polymerization inhibitors, including, but not limited to, taxanes (e.g., paclitaxel and docetaxel), vinca alkaloids (e.g., vinblastine or vinblastine sulfate, vincristine or vincristine sulfate, and vinorelbine), discodermolide, cohesin, and epothilones, and their derivatives. Paclitaxel is commercially available under the trade name Taxol™. Docetaxel is commercially available under the trade name Taxotere™. Vinblastine sulfate is commercially available under the trade name Vinblastin RP™. Vincristine sulfate is commercially available under the trade name Farmistin™.

[0372] The term "alkylating agent" as used herein includes, but is not limited to, cyclophosphamide, ifosfamide, melphalan, or nitrosourea (BCNU or Gliadel). Cyclophosphamide is commercially available under the trade name Cyclostin™. Ifosfamide is commercially available under the trade name Holoxan™.

[0373] The term "histone deacetylase inhibitors" or "HDAC inhibitors" relates to compounds which inhibit histone deacetylase and which possess antiproliferative activity, including, but not limited to, suberoylanilide hydroxamic acid (SAHA).

[0374] The term "antineoplastic antimetabolite" includes, but is not limited to, 5-fluorouracil or 5-FU, capecitabine, gemcitabine, DNA demethylating compounds (e.g., 5-azacytidine and decitabine), methotrexate, and edatrexate, and folate antagonists (e.g., pemetrexed). Capecitabine is commercially available under the trade name Xeloda™. Gemcitabine is commercially available under the trade name Gemzar™.

[0375] The term "platin compound" as used herein includes, but is not limited to, carboplatin, cisplatin, cisplatinum, and oxaliplatin. Carboplatin can be administered, e.g., in the form as it is marketed, e.g., under the trademark Carboplat™. Oxaliplatin can be administered, e.g., in the form as it is marketed, e.g., under the trademark Eloxatin™.

[0376] The term "compounds that target / reduce protein or lipid kinase activity, or protein or lipid phosphatase activity, or further anti-angiogenic compounds" as used herein includes, but is not limited to, protein tyrosine kinase and / or serine and / or threonine kinase inhibitors, or lipid kinase inhibitors, such as a) compounds that target, reduce, or inhibit platelet-derived growth factor-receptor (PDGFR) (e.g., compounds that target or inhibit the activity of PDGFR, in particular compounds that inhibit PDGF receptors, such as N-phenyl-2-pyrimidine-amine derivatives (e.g., imatinib, SU101, SU6668, and GFB-111)), b) compounds that target, reduce, or inhibit fibroblast growth factor receptors (fibroblast growth factor receptors), or c) compounds that target, reduce, or inhibit the activity of PDGFR, in particular compounds that inhibit PDGF receptors, such as N-phenyl-2-pyrimidine-amine derivatives (e.g., imatinib, SU101, SU6668, and GFB-111), c) compounds that target, reduce or inhibit the activity of insulin-like growth factor receptor I (IGF-IR), for example compounds that target, reduce or inhibit the activity of IGF-IR, in particular compounds that inhibit the kinase activity of the IGF-I receptor or antibodies that target the extracellular domain of the IGF-I receptor or its growth factors; d) compounds that target, reduce or inhibit the activity of the Trk receptor tyrosine kinase family or ephrin B4 inhibitors; e) compounds that target, reduce or inhibit the activity of the AxI receptor tyrosine kinase family; f) compounds that target, reduce or inhibit the activity of the Ret receptor tyrosine kinase. g) compounds that target, reduce or inhibit the activity of Kit / SCFR receptor tyrosine kinases (e.g., imatinib); h) compounds that target, reduce or inhibit the activity of C-kit receptor tyrosine kinases (which are part of the PDGFR family), for example, compounds that target, reduce or inhibit the activity of the c-Kit receptor tyrosine kinase family, in particular compounds that inhibit the c-Kit receptor (e.g., imatinib); i) compounds that target, reduce or inhibit the activity of members of the c-Abl family, their gene fusion products (e.g., BCR-Abl kinase), and mutants, for example, compounds that target, reduce or inhibit the activity of c-Abl family members and their gene fusion products;or compounds that inhibit, for example, N-phenyl-2-pyrimidine-amine derivatives (e.g., imatinib or nilotinib (AMN107), PD180970, AG957, NSC 680410, PD173955 from ParkeDavis, or dasatinib (BMS-354825); j) protein kinase C (PKC) and members of the Raf family serine / threonine kinases, MEK, SRC, JAK / pan-JAK, FAK, PDK1, PKB / Akt, Ras / MAPK, PI3K, SYK, TYK2, BTK and members of the TEC family, and / or cyclin-dependent kinases k) compounds which target, reduce or inhibit the activity of members of the protein tyrosine kinase (CDK) family (including staurosporine derivatives such as midostaurin), further examples of compounds include UCN-01, safingol, BAY43-9006, bryostatin 1, perifosine, irmofosine, RO318220 and RO320432, GO6976, lsis3521, LY333531 / LY379196, isoquinoline compounds, FTI, PD184352 or QAN697 (P13K inhibitors) or AT7519 (CDK inhibitors), k) compounds which target, reduce or inhibit the activity of protein tyrosine kinase inhibitors, for example compounds which target, reduce or inhibit the activity of protein tyrosine kinase inhibitors (e.g. mesylate Imatinib acid (Gleevec™) or tyrphostins (e.g., Tyrphostin A23 / RG-50810), AG99, Tyrphostin AG213, Tyrphostin AG1748, Tyrphostin AG490, Tyrphostin B44, Tyrphostin B44 (+) enantiomer, Tyrphostin AG555, AG494, Tyrphostin AG556, AG957, and Adaphostin (4-{[(2,5-dihydroxyphenyl)methyl]amino}-benzoic acid adamantyl ester, NSC680410, Adaphostin), l) compounds which target, decrease or inhibit the activity of the epidermal growth factor family of receptor tyrosine kinases (EGFR1, ErbB2, ErbB3, ErbB4 as homodimers or heterodimers) and their mutants, such as, for exampleCompounds that target, decrease, or inhibit the activity of the epidermal growth factor receptor family, in particular compounds, proteins, or antibodies that inhibit members of the EGF receptor tyrosine kinase family, such as EGF receptor, ErbB2, ErbB3, and ErbB4, or that bind to EGF or an EGF-related ligand (e.g., CP358774, ZD1839, ZM105180), trastuzumab (Herceptin™), cetuximab (Erbitux™), Iressa, Tarceva, OSI-774, Cl-1033, EKB-569, GW-2016, E1.1, E2.4, E2.5, E6.2, E6.4, E2.11, E6.3, or E7.6.3, and 7H-pyrrolo-[2,3-d]pyrimidinone. m) compounds that target, reduce, or inhibit the activity of c-Met, for example, compounds that target, reduce, or inhibit the activity of c-Met, particularly compounds that inhibit the kinase activity of the c-Met receptor, or antibodies that target the extracellular domain of c-Met or bind to HGF; n) compounds that target, reduce, or inhibit the kinase activity of one or more JAK family members (JAK1 / JAK2 / JAK3 / TYK2 and / or pan-JAK) (including, but not limited to, PRT-062070, SB-1578, baricitinib, pacritinib, momelotinib, VX-509, AZD-1480, TG-101348, tofacitinib, and ruxolitinib); o) PI3 kinase (PI3 q) compounds that target, reduce, or inhibit the kinase activity of PI3K (including, but not limited to, ATU-027, SF-1126, DS-7423, PBI-05204, GSK-2126458, ZSTK-474, bupalisib, pictorelisib, PF-4691502, BYL-719, dactolisib, XL-147, XL-765, and idelalisib); q) compounds that target, reduce, or inhibit the signaling effects of the hedgehog protein (Hh) or smoothened receptor (SMO) pathway (including, but not limited to, cyclopamine, vismodegib, itraconazole, erismodegib, and IPI-926 (salidegib));

[0377] As used herein, the term "PI3K inhibitor" includes, but is not limited to, compounds that have inhibitory activity against one or more enzymes in the phosphatidylinositol-3-kinase family (e.g., including, but not limited to, PI3Kα, PI3Kγ, PI3Kδ, PI3Kβ, PI3K-C2α, PI3K-C2β, PI3K-C2γ, Vps34, p110-α, p110-β, p110-γ, p110-δ, p85-α, p85-β, p55-γ, p150, p101, and p87). Examples of PI3K inhibitors useful in the present invention include, but are not limited to, ATU-027, SF-1126, DS-7423, PBI-05204, GSK-2126458, ZSTK-474, bupallisib, pictorelisib, PF-4691502, BYL-719, dactolisib, XL-147, XL-765, and idelalisib.

[0378] The term "Bcl-2 inhibitors" as used herein includes, but is not limited to, compounds having inhibitory activity against B-cell lymphoma 2 protein (Bcl-2) (ABT-199, ABT-731, ABT-737, apogossypol, Ascenta pan-Bcl-2 inhibitors, curcumin (and analogs thereof), dual Bcl-2 / Bcl-xL inhibitors (Infinity Pharmaceuticals / Novartis Pharmaceuticals), Genasense (G3139), HA14-1 (and analogs thereof, see WO 2008 / 118802), navitoclax (and analogs thereof, see U.S. Pat. No. 7,390,799), NH-1 (Shenayng Pharmaceutical University), obatoclax (and analogs thereof, see WO 2004 / 106328), S-001 (Gloria In some embodiments, the Bcl-2 inhibitor is a small molecule therapeutic agent. In some embodiments, the Bcl-2 inhibitor is a peptidomimetic.

[0379] As used herein, the term "BTK inhibitor" includes, but is not limited to, compounds that have inhibitory activity against Bruton's tyrosine kinase (BTK), including, but not limited to, AVL-292 and ibrutinib.

[0380] As used herein, the term "SYK inhibitor" includes, but is not limited to, compounds that have inhibitory activity against spleen tyrosine kinase (SYK) (including, but not limited to, PRT-062070, R-343, R-333, Excellair, PRT-062607, and fostamatinib).

[0381] Further examples of BTK inhibitor compounds and conditions treatable by such compounds in combination with the compounds of the invention can be found in WO 2008 / 039218 and WO 2011 / 090760, which are incorporated by reference in their entireties.

[0382] Further examples of SYK inhibitor compounds and conditions treatable by such compounds in combination with the compounds of the present invention can be found in WO 2003 / 063794, WO 2005 / 007623, and WO 2006 / 078846, each of which is incorporated by reference in its entirety.

[0383] Further examples of PI3K inhibitor compounds and conditions treatable by such compounds in combination with the compounds of the invention can be found in WO 2004 / 019973, WO 2004 / 089925, WO 2007 / 016176, U.S. Pat. No. 8,138,347, WO 2002 / 088112, WO 2007 / 084786, WO 2007 / 129161, WO 2006 / 122806, WO 2005 / 113554, and WO 2007 / 044729, each of which is incorporated by reference in its entirety.

[0384] Further examples of JAK inhibitor compounds and conditions treatable by such compounds in combination with the compounds of the present invention can be found in WO 2009 / 114512, WO 2008 / 109943, WO 2007 / 053452, WO 2000 / 142246, and WO 2007 / 070514, the entireties of which are incorporated herein by reference.

[0385] Additional anti-angiogenic compounds include compounds that have another mechanism of their activity, for example, independent of protein or lipid kinase inhibition, such as thalidomide (Thalomid™) and TNP-470.

[0386] Examples of proteasome inhibitors useful in combination with the compounds of the invention include, but are not limited to, bortezomib, disulfaram, epigallocatechin-3-gallate (EGCG), salinosporamide A, carfilzomib, ONX-0912, CEP-18770, and MLN9708.

[0387] Compounds which target, decrease or inhibit the activity of a protein or lipid phosphatase are eg inhibitors of phosphatase 1, phosphatase 2A or CDC25 (eg okadaic acid or a derivative thereof).

[0388] Compounds that induce cell differentiation processes include, but are not limited to, retinoic acid, α-, γ-, or δ-tocopherol, or α-, γ-, or δ-tocotrienol.

[0389] As used herein, the term cyclooxygenase inhibitor includes, but is not limited to, Cox-2 inhibitors, 5-alkyl substituted 2-arylaminophenylacetic acids and derivatives, such as celecoxib (Celebrex™), rofecoxib (Vioxx™), etoricoxib, valdecoxib, or 5-alkyl-2-arylaminophenylacetic acid (e.g., 5-methyl-2-(2'-chloro-6'-fluoroanilino)phenylacetic acid), lumiracoxib.

[0390] As used herein, the term "bisphosphonate" includes, but is not limited to, etridonic acid, clodronic acid, tiludronic acid, pamidronic acid, alendronic acid, ibandronic acid, risedronic acid, and zoledronic acid. Etridonic acid is commercially available under the trade name Didronel™. Clodronic acid is commercially available under the trade name Bonefos™. Tiludronic acid is commercially available under the trade name Skelid™. Pamidronic acid is commercially available under the trade name Aredia™. Alendronic acid is commercially available under the trade name Fosamax™. Ibandronic acid is commercially available under the trade name Bondranat™. Risedronic acid is commercially available under the trade name Actonel™. Zoledronic acid is commercially available under the trade name Zometa™. The term "mTOR inhibitor" relates to compounds which inhibit the mammalian target of rapamycin (mTOR) and have antiproliferative activity (e.g., sirolimus (Rapamune®), everolimus (Certican™), CCI-779, and ABT578).

[0391] As used herein, the term "heparanase inhibitor" refers to a compound that targets, reduces, or inhibits the degradation of heparin sulfate. This term includes, but is not limited to, PI-88. As used herein, the term "biological response modifier" refers to a lymphokine or interferon.

[0392] As used herein, the term "inhibitors of oncogenic isoforms of Ras," such as H-Ras, K-Ras, or N-Ras, refers to compounds that target, reduce, or inhibit the oncogenic activity of Ras (e.g., "farnesyltransferase inhibitors" such as L-744832, DK8G557, or R115777 (Zarnestra™)). As used herein, the term "telomerase inhibitors" refers to compounds that target, reduce, or inhibit the activity of telomerase. Compounds that target, reduce, or inhibit the activity of telomerase are compounds that inhibit the telomerase receptor (e.g., telomestatin).

[0393] The term "methionine aminopeptidase inhibitor" as used herein refers to a compound that targets, decreases, or inhibits the activity of methionine aminopeptidase. Compounds that target, decrease, or inhibit the activity of methionine aminopeptidase include, but are not limited to, bengamide or a derivative thereof.

[0394] As used herein, the term "proteasome inhibitor" refers to a compound that targets, reduces, or inhibits the activity of the proteasome. Compounds that target, reduce, or inhibit the activity of the proteasome include, but are not limited to, bortezomib (Velcade™) and MLN341.

[0395] As used herein, the term "matrix metalloproteinase inhibitors" or ("MMP" inhibitors) includes, but is not limited to, collagen peptidomimetic and non-peptidomimetic inhibitors, tetracycline derivatives (e.g., the hydroxamate peptidomimetic inhibitor batimastat), and its orally bioavailable analogs marimastat (BB-2516), prinomastat (AG3340), metastat (NSC683551), BMS-279251, BAY12-9566, TAA211, MMI270B, or AAJ996.

[0396] As used herein, the term "compounds used in the treatment of hematological malignancies" includes, but is not limited to, FMS-like tyrosine kinase inhibitors, which are compounds that target, decrease, or inhibit the activity of FMS-like tyrosine kinase receptor (Flt-3R). Interferon, 1-β-D-arabinofuransylcytosine (ara-c), and bisulfan, and ALK inhibitors are compounds that target, decrease, or inhibit anaplastic lymphoma kinase.

[0397] Compounds that target, decrease, or inhibit the activity of FLT-3R are, in particular, compounds, proteins, or antibodies that inhibit members of the Flt-3R receptor kinase family (e.g., PKC412, midostaurin, staurosporine derivatives, SU11248, and MLN518).

[0398] The term "HSP90 inhibitor," as used herein, includes, but is not limited to, compounds that target, reduce, or inhibit the intrinsic ATPase activity of HSP90 and target, reduce, or inhibit HSP90 client proteins for degradation via the ubiquitin proteosome pathway. Compounds that target, reduce, or inhibit the intrinsic ATPase activity of HSP90 are, in particular, compounds, proteins, or antibodies that inhibit the ATPase activity of HSP90 (e.g., 17-allylamino, 17-demethoxygeldanamycin (17AAG), geldanmycin derivatives, other geldanmycin-related compounds, radicicol, and HDAC inhibitors).

[0399] The term "antiproliferative antibody" as used herein includes, but is not limited to, trastuzumab (Herceptin™), trastuzumab-DM1, Erbitux, bevacizumab (Avastin™), rituximab (Rituxan®), PRO64553 (anti-CD40), and 2C4 antibodies. Antibody refers to intact monoclonal antibodies, polyclonal antibodies, multispecific antibodies formed from at least two intact antibodies, and antibody fragments, so long as they exhibit the desired biological activity.

[0400] For the treatment of acute myeloid leukemia (AML), the compounds of the present invention can be used in combination with standard leukemia therapies (especially in combination with therapeutic agents used to treat AML). In particular, the compounds of the present invention can be administered in combination with, for example, farnesyltransferase inhibitors and / or other drugs useful in the treatment of AML (e.g., daunorubicin, adrimycin, Ara-C, VP-16, teniposide, mitoxantrone, idarubicin, carboplatinum, and PKC412).

[0401] Other anti-leukemia compounds include, for example, Ara-C, pyrimidine analogs, which are 2'-α-hydroxyribose (arabinoside) derivatives of deoxycytidine. Also included are the purine analogs hypoxanthine, 6-mercaptopurine (6-MP), and fludarabine phosphate. Compounds that target, decrease, or inhibit the activity of histone deacetylase (HDAC) inhibitors, such as sodium butyrate and suberoylanilide hydroxamic acid (SAHA), inhibit the activity of enzymes known as histone deacetylases. Specific HDAC inhibitors include MS275, SAHA, FK228 (formerly FR901228), trichostatin A, and compounds disclosed in U.S. Pat. No. 6,552,065 (including, but not limited to, N-hydroxy-3-[4-[[2-(2-methyl-1}H-indol-3-yl)-ethyl]-amino]methyl]phenyl]-2E-2-propenamide, or a pharmaceutically acceptable salt thereof, and N-hydroxy-3-[4-[(2-hydroxyethyl){2-(1H-indol-3-yl)-ethyl]-amino]methyl]phenyl]-2E-2-propenamide, or a pharmaceutically acceptable salt thereof (particularly the lactate salt)). As used herein, somatostatin receptor antagonist refers to a compound that targets or inhibits somatostatin receptors (e.g., octreotide and SOM230). Approaches that damage tumor cells refer to approaches such as ionizing radiation. The term "ionizing radiation" referred to above and below means ionizing radiation that occurs as electromagnetic rays (e.g., X-rays and gamma rays) or particles (e.g., alpha particles and beta particles). Ionizing radiation is provided in radiation therapy and is known in the art, but is not limited to this (see Hellman, Principles of Radiation Therapy, Cancer, in Principles and Practice of Oncology, Devita et al., Eds., 4th Edition, Vol. 1, pp. 248-275 (1993)).

[0402] Also included are EDG binders and ribonucleotide reductase inhibitors. As used herein, the term "EDG binder" refers to a class of immunosuppressants that regulate lymphocyte recirculation (e.g., FTY720). The term "ribonucleotide reductase inhibitor" refers to a pyrimidine or purine nucleoside analog, including, but not limited to, fludarabine and / or cytosine arabinoside (ara-C), 6-thioguanine, 5-fluorouracil, cladribine, 6-mercaptopurine (especially in combination with ara-C for ALL), and / or pentostatin. Ribonucleotide reductase inhibitors are, in particular, hydroxyurea or 2-hydroxy-1H-isoindole-1,3-dione derivatives.

[0403] Also included are, inter alia, VEGF compounds, proteins, or monoclonal antibodies, such as 1-(4-chloroanilino)-4-(4-pyridylmethyl)phthalazine, or a pharmaceutically acceptable salt thereof, 1-(4-chloroanilino)-4-(4-pyridylmethyl)phthalazine succinate; Angiostatin™; Endostatin™; anthranilic acid amide; ZD4190; ZD6474; SU5416; SU6668; bevacizumab; or anti-VEGF antibodies or anti-VEGF receptor antibodies such as rhuMAb and RHUFab, VEGF aptamers (e.g., Macugon); FLT-4 inhibitors, FLT-3 inhibitors, VEGFR-2 IgGI antibodies, Angiozyme (RPI4610), and bevacizumab (Avastin™).

[0404] As used herein, photodynamic therapy refers to therapy that uses certain chemicals known as photosensitizing compounds to treat or prevent cancer. Examples of photodynamic therapy include treatment with compounds such as Visudyne™ and porfimer sodium.

[0405] As used herein, angiogenic antisteroids refer to compounds that block or inhibit angiogenesis (e.g., anecortave, triamcinolone, hydrocortisone, 11-α-epihydrocortisol, cortexolone, 17α-hydroxyprogesterone, corticosterone, desoxycorticosterone, testosterone, estrone, and dexamethasone).

[0406] Corticosteroid-containing implants refer to compounds such as fluocinolone and dexamethasone.

[0407] Other chemotherapeutic compounds include, but are not limited to, plant alkaloids, hormonal compounds and hormone antagonists, biological response modifiers (preferably lymphokines or interferons), antisense oligonucleotides or oligonucleotide derivatives, shRNA or siRNA, or other compounds, or compounds with other or unknown mechanisms of action.

[0408] The structures of active compounds identified by code number, generic name or trade name can be obtained from the actual edition of the standard compendium "The Merck Index" or from databases such as Patents International (e.g. IMS World Publications).

[0409] The compounds of the present invention can also be used in combination with known therapeutic processes (e.g., administration of hormones or radiation). In certain embodiments, provided compounds are used as radiosensitizers, particularly for treating tumors that are poorly sensitive to radiation therapy.

[0410] The compounds of the present invention can be administered alone or in combination with one or more other therapeutic compounds, and possible combination therapies can be in the form of a fixed combination, or in the form of a compound of the present invention and one or more other therapeutic compounds administered alternately or independently, or in the form of a fixed combination administered in combination with one or more other therapeutic compounds.The compounds of the present invention can also be administered in combination with chemotherapy, radiotherapy, immunotherapy, phototherapy, surgical intervention, or a combination thereof, particularly for tumor therapy.As mentioned above, long-term therapy is also possible, as well as adjuvant therapy in conjunction with other treatment strategies.Other possible therapies are therapy to maintain the patient's condition after tumor regression, or even, for example, chemoprevention therapy in at-risk patients.

[0411] These additional agents can be administered separately from the composition containing the compound of the present invention as part of a multiple dose regimen. Alternatively, these agents can be part of a single dosage form, mixed together with the compound of the present invention in a single composition. When administered as part of a multiple dose regimen, the two active agents can be provided simultaneously, sequentially, or within a period of each other, usually within 5 hours of each other.

[0412] As used herein, the terms "combination," "combined," and related terms refer to simultaneous or sequential administration of therapeutic agents according to the invention. For example, a compound of the invention may be administered with another therapeutic agent, either simultaneously or sequentially, in separate unit dosage forms, or together in a single unit dosage form. Thus, the invention provides a single unit dosage form comprising a compound of the invention, an additional therapeutic agent, and a pharmaceutically acceptable carrier, adjuvant, or vehicle.

[0413] The amounts of both the compounds of the present invention and additional therapeutic agents (in compositions containing such additional therapeutic agents) that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated and the particular mode of administration. Preferably, the compositions of the present invention should be formulated so that a dosage of 0.01 to 100 mg / kg body weight / day of the compound of the present invention can be administered.

[0414] In those compositions that include an additional therapeutic agent, the additional therapeutic agent and the compound of the invention may act synergistically. Thus, the amount of additional therapeutic agent in such compositions will be less than the amount required in a monotherapy utilizing only that therapeutic agent. In such compositions, dosages of 0.01 to 1,000 μg / kg body weight / day of the additional therapeutic agent can be administered.

[0415] The amount of additional therapeutic agent present in the compositions of the invention will be less than the amount that would normally be administered in a composition comprising that agent as the only active agent. Preferably, the amount of additional therapeutic agent in the compositions of the present disclosure will be in the range of about 50% to 100% of the amount that would normally be present in a composition comprising that agent as the only therapeutically active agent.

[0416] In some embodiments, the additional therapeutic agent administered in combination with the compound of the present invention is another mTOR inhibitor. In some embodiments, the additional mTOR inhibitor inhibits mTOR by binding to the catalytically active site of mTOR. Examples of such additional mTOR inhibitors include dactolisib, 8-(6-methoxy-pyridin-3-yl)-3-methyl-1-(4-piperazin-1-yl-3-trifluoromethyl-phenyl)-1,3-dihydro-imidazo[4,5-c]quinolin-2-one (WO 2006 / 122806), bistusertib (AZD2014; WO 2009 / 153597), AZD8055. (WO 2009 / 153597, XL388 (U.S. Patent Application Publication No. 2010 / 0305093), sapanisertib (MLN0128; INK128, WO 2015 / 051043), DS3078, apitolisib (GDC0980; WO 2008 / 070740), omipalisib (GSK-2126458; WO 2008 / 14446), NVP-BGT226 (Chang, KY, et al., Clin. Cancer Res.17(22):7116-26(2011)), voxtalisib (XL765; SAR245409, International Publication No. 2007 / 044813), PF04691502 (International Publication No. 2008 / 032162), jedatolicib (PF05212384; PKI-587, International Publication No. 2009 / 143313), SF1126 (International Publication No. Publication No. 2004 / 089925), GSK1059615 (International Publication No. 2007 / 136940), BI-860585, OSI027 (International Publication No. 2007 / 061737), VS5584 (International Publication No. 2010 / 114484), CC-223 (International Publication No. 2010 / 062571), DCBCI-0901 (Lee, YE, et al. al., Mol. Canc. Thera. 12(11Suppl):Abstract nr C270(2013)):), LY3023414 (WO 2012 / 097039), P529 (WO 2007 / 133249), panulisib (P7170; WO 2012 / 007926), DS-7423 (Kashiyama, T., et al.,PLoS One9(2):e87220(2014)), PWT33567 mesylate (VCD-597; International Publication No. 2010 / 110685), ME-344 (NV-128; Navarro, P., et al.,Cell Rep.15(12):2705-18(2016)), ABTL0812(International Publication No. 2010 / 106211), WYE-132, EXEL-3885(Eur J Cancer Suppl.6(12):Abst 322(2008)), EXEL-4431(Eur J Cancer Suppl.6(12):Abst 322(2008)), AR-mTOR-26(101st Annu Meet Am Assoc Cancer Res(AACR)(April 17-21, Washington, DC) 2010, Abst 4484), NV-128 (AB Alvero et al., Mol Cancer Ther. 10(8):1385-93(2011)), salinomycin (VS-507; Gupta, P.B., et al., Cell 138(4):645-59(2009)), BN-107, BN-108, WAY-600, WYE-687, WYE-354 (Yu, K., et al., Cancer Res. 69(15):6232-40(2009)), Ku-063794 (Garcia-Martinez, J.M., et al.,Biochem.J.421(1):29-42(2009)), Torquinib (PP242;Apsel,B.,et al., Nat.Chem.Biol.4(11):691-99(2008)), PP30, CZ415(REF), INK1069, EXEL-2044, EXEL-7518, SB2158, SB2280, AR-mTOR-1(Wallace,EM,et al.,Mol.Canc.Thera.8(12 Suppl):Abst.B267(2009)). .

[0417] Reference herein to any specific additional mTOR inhibitor also includes any pharmaceutically acceptable salts, stereoisomers, tautomers, solvates, hydrates, and polymorphs thereof.

[0418] The compounds of the present invention, or pharmaceutical compositions thereof, can also be incorporated into compositions for coating implantable medical devices, such as prostheses, artificial valves, vascular grafts, stents, and catheters. For example, vascular stents are used to overcome restenosis (re-narrowing of the blood vessel wall after injury). However, patients using stents or other implantable devices are at risk of clot formation or platelet activation. These undesirable effects can be prevented or mitigated by pre-coating the device with a pharmaceutically acceptable composition containing a kinase inhibitor. An implantable device coated with the compounds of the present invention is another embodiment of the present invention.

[0419] Example As shown in the examples below, in certain exemplary embodiments, compounds are prepared according to the following general procedures. The general methods represent the synthesis of specific compounds of the present invention, the following general methods, and other methods known to those skilled in the art, but are understood to be applicable to all compounds, and each subclass and species of these compounds, as described herein (see Luengo, JI et al., Chem. Biol., 2(7): 471-81 (1995); Grinfeld, AA et al., Tet. Lett., 35(37): 6835-38 (1994); International Application No. PCT / US2019 / 037507 and International Application No. PCT / US2020 / 063351, which are incorporated herein by reference in their entirety).

[0420] The following examples are based on LC / MS, 1 H NMR, 19 Where only analytical measurements such as F NMR are listed (rather than details of reaction steps), it is understood that the title compound was prepared according to the procedures set forth in the synthetic schemes and examples herein, selecting and substituting suitable reagents and reactants as would be readily recognized by one of ordinary skill in the art.

[0421] Unless otherwise indicated in the examples, all temperatures are in degrees Celsius (°C). All reactions were carried out under an inert atmosphere at ambient temperature unless otherwise noted. Unless otherwise noted, reaction solutions were stirred at room temperature under an N2(g) or Ar(g) atmosphere. Reagents used without synthetic details are commercially available or prepared according to known methods, e.g., literature procedures. When solutions are "concentrated to dryness," they are concentrated using a rotary evaporator under reduced pressure, and when solutions are dried, they are typically dried over a desiccant such as MgSO4 or Na2SO4. When a synthetic product is described as being isolated as a residue, the term "residue" is not intended to limit the physical state from which the product is isolated and may include, for example, a solid, oil, foam, gum, syrup, etc., as will be understood by those skilled in the art.

[0422] In obtaining the compounds described in the Examples below and the corresponding analytical data, the following experimental and analytical protocols were followed unless otherwise indicated.

[0423] LC-MS: Unless otherwise indicated, the analytical LC-MS system used consisted of a Shimadzu LCMS-2020 with electrospray ionization (ESI) in positive ion detection mode, equipped with a 20ADXR pump, a SIL-20ACXR autosampler, a CTO-20AC column oven, an M20A PDA detector, and an LCMS 2020 MS detector. The column was a HALO C18 30 *The column diameter was 5.0 mm, 2.7 μm. Mobile phase A was water containing 0.05% TFA, and mobile phase B was acetonitrile containing 0.05% TFA. The gradient was 5% to 100% (95%) mobile phase B over 2.0 min, held for 0.7 min, then back to 5% mobile phase B over 0.05 min, held for 0.25 min. The column oven (CTO-20AC) was operated at 40.0 °C. The flow rate was 1.5 mL / min, and the injection volume was 1 μL. PDA (SPD-M20A) detection was in the range of 190–400 nm. The MS detector was configured for electrospray ionization as the ionization source, acquisition mode: scan, nebulization gas flow rate: 1.5 L / min, drying gas flow rate: 15 L / min, detector voltage: tuning voltage ±0.2 kV, DL temperature: 250 °C, heat block temperature: 250 °C, scan range: 90.00 to 900.00 m / z. ELSD (Alltech 3300) detector parameters: drift tube temperature: 60 ±5 °C; N2 flow rate: 1.8 ±0.2 L / min. The mobile phase gradient was optimized for each compound. Calculated masses correspond to exact masses.

[0424] Preparative HPLC: Unless otherwise noted, preparative HPLC purifications were performed using a Waters Auto Purification System (2545-2767) equipped with a 2489 UV detector. The column was selected from one of the following: Waters C18, 19 x 150 mm, 5 μm; XBridge Prep OBD C18 column, 30 x 150 mm, 5 μm; XSelect CSH Prep C18 OBD column, 5 μm, 19 * The columns were: 150 mm, XBridge Shield RP18 OBD column, 30 x 150 mm, 5 μm; Xselect CSH Fluorophenyl, 30 x 150 mm, 5 μm; or YMC-Actus Triart C18, 30 x 150 mm, 5 μm. The mobile phase consisted of a mixture of acetonitrile (5–95%) in water containing 0.1% FA or 10 mmol / L NH4HCO3. The flow rate was maintained at 25 mL / min, the injection volume was 1200 μL, and two UV detector channels were used: 254 nm and 220 nm. The mobile phase gradient was optimized for each compound.

[0425] Normal-phase flash chromatography: Unless otherwise noted, normal-phase flash column chromatography (FCC) was performed on silica gel in prepackaged silica gel columns (such as RediSep®) using ethyl acetate (EtOAc) / hexane, ethyl acetate (EtOAc) / petroleum ether (bp 60-90°C), CHCl / MeOH, or 10% 2N NH in CHCl / MeOH as the eluent.

[0426] 1 H NMR: unless otherwise noted 1 H NMR spectra were acquired using a 400 MHz spectrometer (or a 500 MHz spectrometer) in DMSO-d6 or CDCl3 solutions. Nuclear magnetic resonance (NMR) spectral characteristics refer to chemical shifts (δ) and are expressed in parts per million (ppm). Tetramethylsilane (TMS) was used as an internal standard in DMSO-d6 solutions. Coupling constants (J) are reported in hertz (Hz). The nature of the shifts relative to multiplicity is reported as s (singlet), d (doublet), t (triplet), q (quartet), dd (double doublet), dt (double triplet), m (multiplet), or br (broad).

[0427] List of abbreviations used in the experimental section. CH3CN:Acetonitrile DCM: dichloromethane DMAP: dimethylaminopyridine DMF: N,N-dimethylformamide DMSO: dimethyl sulfoxide EDCl: 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide ESI: electrospray ionization EtOAc: ethyl acetate Et2O: Diethyl ether EtOH: ethanol h: time HCl: Hydrogen chloride HF: Hydrogen fluoride HND-8: Acidic ion exchange resin (e.g., Amberlyst) H2O: Water HPLC: High-Performance Liquid Chromatography MeOH: Methanol min:minutes MgSO4: Magnesium sulfate mL: milliliter mM: millimolar mmol: millimolar MS: Mass spectrometry N2: Nitrogen gas NaHCO3: Sodium bicarbonate NaOH: Sodium hydroxide Na2SO4: Sodium sulfate NH3: Ammonia NH4Cl: Ammonium chloride NMR: nuclear magnetic resonance ℃: degrees (Celsius) prep-HPLC: Preparative High-Performance Liquid Chromatography PE: Petroleum ether p-TsOH: para-toluenesulfonic acid rt: room temperature TASF: Tris(dimethylamino)sulfonium difluorotrimethylsilicic acid TEA: Triethylamine TFA: Trifluoroacetic acid THF: tetrahydrofuran

[0428] Synthesis Example: Intermediates Synthesis of intermediate I

[0429] [ka]

[0430] To a solution of rapamycin (0.2 g, 0.22 mmol) in toluene (5 mL) was added proton sponge (0.94 g, 4.38 mmol) at room temperature, followed by methyl trifluoromethanesulfonate (0.54 g, 3.28 mmol). The mixture was stirred at 50°C for 6 h, then cooled and purified by silica gel chromatography and reverse-phase chromatography (85% CH3CN in water) to give (24E, 26E, 28E, 29E, 31R, 32S, 33R, 34R, 36S, 38S, 40S, 41S, 42R, 43R, 52R)-41-[(1R)-2-[(1S,3R,4R)-3,4-dimethoxycyclohexane]. [Hexyl]-1-methyl-ethyl]-52-hydroxy-40,42,43-trimethoxy-31,32,33,34,44,45-hexamethyl-60,61-dioxa-53-azatricyclohexatriaconta-24,26,28(44),29(45)-tetraene-46,47,48,49,50-pentone (50 mg, 24% yield) was obtained as a white solid. ESI-MS (EI + , m / z): 964.2 [M + Na] + . 1 H NMR(400MHz,CDCl3)δ 6.50-5.80(m,4H),5.62(ddd,J=22.9,14.5,7.9Hz,1H),5.32(dt,J=11.6,7.7Hz,2H),5.1 8-5.03(m,1H),4.68(s,1H),3.95-3.54(m,5H),3.50-3.33(m,7H),3.32-3.21(m,3H),3.18 -2.92(m,8H),2.83-2.48(m,3H),2.25(dd,J=30.1,10.7Hz,2H),2.02(ddd,J=34.0,26.3, 9.6Hz, 4H), 1.88-1.56 (m, 14H), 1.51-1.16 (m, 9H), 1.15-0.82 (m, 18H), 0.79-0.68 (m, 1H).

[0431] Synthesis of Intermediate II

[0432] [ka]

[0433] Step 1: 3-iodopropyl trifluoromethanesulfonate. A mixture of 3-iodopropan-1-ol (4 g, 21.51 mmol) and 2,6-lutidine (4.61 g, 43 mmol) in DCM (40 mL) was cooled to 0 °C under N2, and trifluoromethylsulfonyl trifluoromethanesulfonate (6.67 g, 23.66 mmol) was added dropwise. The resulting solution was stirred at 0 °C for 2 h, then quenched with 10% EtOAc in PE and filtered through a short silica gel column. The filtrate was concentrated in vacuo to give 3-iodopropyl trifluoromethanesulfonate (6.72 g, 98% yield) as a pale yellow liquid.

[0434] Step 2: (22E,24E,26E,27E,32R,33S,34R,35R,37S,39S,41S,42S,43R,44R,53R)-43,53-dihydroxy-42-[(1R)-2-[(1S,3R,4R)-4-(3-iodopropoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-41,44-dimethoxy-32,33,34,35,45,46-hexamethyl-62,63-dioxa-54-azatricyclohexatriaconta-22,24,26(45),27(46)-tetraene-47,48,49,50,51-pentone (Intermediate II). A mixture of rapamycin (2 g, 2.19 mmol) and N-ethyl-N-isopropyl-propan-2-amine (5.72 mL, 32.82 mmol) in toluene (40 mL) was stirred at 50 °C for 16 hours. The mixture was poured into ice-cold saturated aqueous NaHCO (50 mL), washed with ice water (60 mL × 2), brine (50 mL), dried over anhydrous NaSO, filtered, and concentrated. The residue was purified by silica gel chromatography (PE:EA = 3:1) to give intermediate II (1.45 g, 60% yield) as a pale yellow solid. ESI-MS (EI+, m / z): 1104.5 [M+Na] + .

[0435] Synthesis of Intermediates III and VI

[0436] [ka]

[0437] Step 1: 2-Methoxyethyl trifluoromethanesulfonate. To a solution of 2-methoxyethanol (4.5 g, 59.14 mmol) and DIEA (11.46 g, 88.71 mmol) in DCM (50 mL) was added trifluoromethylsulfonyl trifluoromethanesulfonate (18.35 g, 65.05 mmol) dropwise at 0° C. under N. This was stirred at 0° C. for 2 hours, then diluted with DCM (50 mL) and washed with saturated NaHCO (50 mL), water (50 mL), and brine (50 mL). The organic layer was then dried over NaSO, filtered, and concentrated in vacuo to give 2-methoxyethyl trifluoromethanesulfonate (12.3 g, 99% yield) as a brown oil, which was used without further purification. 1 H NMR (400MHz, CDCl3): δ 4.62 (t, J = 4.4 Hz, 2H), 3.71 (t, J = 4.6 Hz, 2H), 3.42 (s, 3H).

[0438] Step 2: (3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-9,27-dihydroxy-10,21-dimethoxy-3-((R)-1-((1S,3R,4R)-3-methoxy-4-(2-methoxyethoxy)cyclohexyl)propan-2-yl)-6,8,12, 14,20,26-Hexamethyl-9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-hexadecahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohentriacontin-1,5,11,28,29(4H,6H,31H)-pentane (Intermediate III). Rapamycin (3 g, 3.28 mmol) and N-ethyl-N-isopropyl-propan-2-amine (8.48 g, 65.63 mmol) in toluene (30 mL) were stirred at 50 °C for 3 h. The reaction mixture was poured into cold NaHCO (50 mL), washed with ice water (2 × 60 mL), brine (50 mL), dried over anhydrous NaSO, filtered, and concentrated. The residue was purified by silica gel chromatography (PE:EA = 1:2) to give intermediate III (1.2 g, 38% yield) as a white solid. 1 H NMR(400MHz,CDCl3):δ 5.95-6.42(m,4H),5.14-5.58(m,4H),4.41-4.81(m,1H),4.17-4.28(m,1H),3.84 -4.00(m,1H),3.63-3.79(m,4H),3.49-3.59(m,2H),3.31-3.46(m,10H),3.07-3.2 2(m,5H),2.55-2.76(m,2H),2.31-2.35(m,1H),1.91-2.10(m,3H),1.61-1.88(m,1 9H), 1.41-1.55(m, 4H), 1.15-1.36(m, 7H), 0.83-1.11(m, 16H), 0.69-0.76(m, 1H).

[0439] Step 3: (3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-27-hydroxy-9,10,21-trimethoxy-3-((R)-1-((1S,3R,4R)-3-methoxy-4-(2-methoxyethoxy)cyclohexyl)propan-2-yl)-6,8,12, 14,20,26-Hexamethyl-9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-hexadecahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohentriacontin-1,5,11,28,29(4H,6H,31H)-pentaone (intermediate VI). (23E,25E,27E,28E,32R,33S,34R,35R,37S,39S,41S,42S,43R,44R,53R)-43,53-dihydroxy-41,44-dimethoxy-42-[(1R)-2-[(1S,3R,4R)-3-methoxy-4-(2-methoxyethoxy)cyclohexyl]-1-methyl-ethyl]-32,33,34,35,45,46-hexamethoxy in toluene (10 mL) To a suspension of ethyl-62,63-dioxa-54-azatricyclohexatriaconta-23,25,27(45),28(46)-tetraene-47,48,49,50,51-pentone (0.5 g, 0.51 mmol) and 1,8-bis(dimethylamino)naphthalene (1.65 g, 7.71 mmol) was added methyl trifluoromethanesulfonate (1.01 g, 6.17 mmol) dropwise at room temperature under N. The reaction was heated to 50 °C for 3 h, then filtered, diluted with EA (60 mL), and washed with saturated aqueous NH.sub.4Cl (60 mL × 10), water (60 mL), and brine (60 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel chromatography (PE:EA=1:1) to give intermediate VI (92 mg, 18% yield) as a white solid. 1H NMR (400MHz, CDCl3): δ 6.03-6.42(m,4H),5.08-5.60(m,4H),4.10-4.74(m,1H),3.73-3.93(m,4H),3.49 -3.71(m,5H),3.44-3.46(m,3H),3.34-3.41(m,4H),3.24-3.31(m,3H),3.07-3.18 (m,7H),2.48-2.82(m,2H),1.95-2.35(m,5H),1.53-1.83(m,18H),1.42-1.52(m, 3H),1.22-1.37(m,6H),1.04-1.15(m,10H),0.86-0.97(m,7H),0.69-0.79(m,1H).

[0440] Synthesis of Intermediate IV

[0441]

change

[0442] Step 1: (3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-3-((R)-1-((1S,3R,4R)-4-((tert-butyldimethylsilyl)oxy)-3-methoxycyclohexyl)propan-2-yl)-9,27-dihydroxy-10,21-dimethoxy To a solution of rapamycin (5 g, 5.47 mmol) in DMF (60 mL) was added imidazole (1.49 g, 21.88 mmol) and tert-butyl-chloro-dimethyl-silane (2.47 g, 16.41 mmol, 3.05 mL). The reaction was stirred at 50 °C for 6 h and then poured into ice-cold saturated NH4Cl solution (40 mL) and Et2O:PE (60 mL, 2:1). The organic layer was washed with saturated NH4Cl solution (20 mL), water, and brine (20 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel chromatography (10% to 50% EtOAc in PE) to give (3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-3-((R)-1-((1S,3R,4R)-4-((tert-butyldimethylsilyl)oxy)-3-methoxycyclohexyl)propan-2-yl)-9,27-dihydroxy-10 ,21-Dimethoxy-6,8,12,14,20,26-hexamethyl-9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-hexadecahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohentriacontin-1,5,11,28,29(4H,6H,31H)-pentane (4 g, 71% yield) was obtained as a white solid. ESI-MS (EI + , m / z): 1050.5 [M + Na] + .

[0443] Step 2: (3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-3-((R)-1-((1S,3R,4R)-4-((tert-butyldimethylsilyl)oxy)-3-methoxycyclohexyl)propan-2-yl)-27-hydroxy-9,10,21-trimethoxysilane Ci-6,8,12,14,20,26-hexamethyl-9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-hexadecahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohentriacontin-1,5,11,28,29(4H,6H,31H)-pentaone. (3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-3-((R)-1-((1S,3R,4R)-4-((tert-butyldimethylsilyl)oxy)-3-methoxycyclohexyl)propan-2-yl)-9,27-dihydroxy-10,21-dimethoxy-6,8,12,14,20,26-hexamethyl-9,10,12,13,14,21,22,23,24,25 To a suspension of 5,26,27,32,33,34,34a-hexadecahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohentriacontin-1,5,11,28,29(4H,6H,31H)-pentaone (1 g, 0.97 mmol) and 1,8-bis(dimethylamino)naphthalene (2.5 g, 11.67 mmol), methyl trifluoromethanesulfonate (2.39 g, 14.59 mmol, 1.60 mL) was added dropwise at room temperature under N. The reaction was then heated to 50 °C for 6 h, cooled, and filtered.The filtrate was concentrated and purified by silica gel chromatography to give (28E,30E,32E,33E,35R,36S,37R,38R,40S,42S,44S,45S,46R,47R,56R)-45-[(1R)-2-[(1S,3R,4R)-4-[tert-butyl(dimethyl)silyl]oxy-3-methoxy-cyclohexyl]-1-methyl [N-ethyl]-56-hydroxy-44,46,47-trimethoxy-35,36,37,38,48,49-hexamethyl-65,66-dioxa-58-azatricyclohexatriaconta-28,30,32(48),33(49)-tetraene-50,51,52,53,54-pentone (0.45 g, 44% yield) was obtained as a white solid. ESI-MS (EI). + , m / z): 1064.6 [M + Na] + .

[0444] Step 3: (3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-27-hydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propan-2-yl)-9,10,21-trimethoxy-6,8,12, 14,20,26-Hexamethyl-9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-hexadecahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohentriacontin-1,5,11,28,29(4H,6H,31H)-pentaone. (3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-3-((R)-1-((1S,3R,4R)-4-((tert-butyldimethylsilyl)oxy)-3-methoxycyclohexyl)propan-2-yl)-27-hydroxy-9,10,21-trimethoxy-6,8,12,14,20,26-hexamethyl-9,10,12, To a solution of 13,14,21,22,23,24,25,26,27,32,33,34,34a-hexadecahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohentriacontin-1,5,11,28,29(4H,6H,31H)-pentaone (0.4 g, 0.38 mmol) was added pyridine hydrofluoride (3.8 g, 38.37 mmol) at 0 °C, and the mixture was stirred at 45 °C for 5 h. The mixture was diluted with DCM and saturated NaHCO, washed with water and brine, dried over NaSO, filtered, and concentrated.The residue was purified by reverse phase chromatography to give (3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-27-hydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propan-2-yl)-9,10,21-trimethoxy-6,8,12,14, 20,26-Hexamethyl-9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-hexadecahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohentriacontin-1,5,11,28,29(4H,6H,31H)-pentaone (0.16 g, 45% yield) was obtained as a white solid. ESI-MS (EI). + , m / z): 949.9 [M + Na] + .

[0445] Step 4: (1R,2R,4S)-4-((R)-2-((3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-27-hydroxy-9,10,21-trimethoxy-6,8,12,14,20,26-hexamethyl-1,5,11,28,29-pentaoxo-1,4,5,6 ,9,10,11,12,13,14,21,22,23,24,25,26,27,28,29,31,32,33,34,34a-Tetracosahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohentriacontin-3-yl)propyl)-2-methoxycyclohexyldimethylphosphinate (Intermediate IV). (3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-27-hydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propan-2-yl)-9,10,21-trimethoxy-6,8,12,14,20,26-hexamethyl-9,10,12,13,14,21,22,23,24,25,26,27,3 To a solution of 2,33,34,34a-hexadecahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohentriacontin-1,5,11,28,29(4H,6H,31H)-pentaone (0.26 g, 0.28 mmol) in DCM (1 mL) was added 2,6-di-tert-butyl-4-methylpyridine (0.17 g, 0.84 mmol) and dimethylphosphinic chloride (0.315 g, 2.80 mmol) at 0° C. The reaction was stirred at 0° C. for 5 h, then diluted with EtOAc, washed with saturated NaHCO solution, ice-cold 0.5 N HCl, water, brine, dried over NaSO, filtered, and concentrated. The residue was purified by silica gel chromatography (DCM:MeOH=40:1) to give intermediate IV (100 mg, 36% yield) as a white solid. + , m / z): 1025.8 [M + Na] + .

[0446] Synthesis of intermediate V / IX

[0447] [ka]

[0448] Step 1: 2-((tert-butyldiphenylsilyl)oxy)ethan-1-ol. tert-Butylchlorodiphenylsilane (26.61 g, 96.83 mmol) was added to a solution of ethylene glycol (49.28 g, 793.97 mmol) in pyridine (44 mL) at 0 °C. The resulting solution was stirred at room temperature for 1 h, then poured into water (500 mL) and extracted with EtOAc (200 mL × 3). The organic layer was dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel chromatography (EA:PE = 1:8) to give 2-((tert-butyldiphenylsilyl)oxy)ethan-1-ol (25 g, 86% yield) as a colorless solid. ESI-MS (EI + , m / z): 323.1 [M + Na] + .

[0449] Step 2: 2-((tert-Butyldiphenylsilyl)oxy)ethyl trifluoromethanesulfonate. To a solution of 2-((tert-butyldiphenylsilyl)oxy)ethan-1-ol (17.13 g, 57 mmol) and DIEA (11.05 g, 85.52 mmol) in DCM (120 mL) was added trifluoromethylsulfonyl trifluoromethanesulfonate (17.69 g, 62.71 mmol) at 0° C. under N. The reaction was stirred at 0° C. for 2 h, then diluted with DCM (200 mL) and washed with saturated NaHCO (100 mL × 3), water (100 mL × 2), and brine (100 mL). The organic layer was dried over NaSO, filtered, and concentrated to give 2-((tert-butyldiphenylsilyl)oxy)ethyl trifluoromethanesulfonate (24.5 g, 99% yield) as a brown oil, which was used without further purification.

[0450] Step 3: (35E,37E,39E,40E,46R,47S,48R,49R,51S,53S,55S,56S,57R,58R,67R)-56-[(1R)-2-[(1S,3R,4R)-4-[2-[tert-butyl(diphenyl)silyl]oxyethoxy]-3-methoxy-cyclohexyl]-1-methyl-ethyl]-57,67-dihydroxy-55,58-dimethoxy-46,47,48,49,59,60-hexamethyl-77,78-dioxa-69-azatricyclohexatriaconta-35,37,39(59),40(60)-tetraene-61,62,63,64,65-pentone. To a solution of rapamycin (5 g, 5.47 mmol) and 2-((tert-butyldiphenylsilyl)oxy)ethyl trifluoromethanesulfonate (23.66 g, 54.69 mmol) in toluene (100 mL) was added DIEA (8.48 g, 65.63 mmol). The mixture was stirred at 58 °C for 16 h, then poured into ice-cold NaHCO solution (200 mL) and extracted with EtOAc (100 mL × 3). The organic layer was washed with water (100 mL × 3), brine (100 mL), dried over anhydrous NaSO, filtered, and concentrated. The residue was purified by silica gel chromatography (PE:EA=3:1) to give (35E,37E,39E,40E,46R,47S,48R,49R,51S,53S,55S,56S,57R,58R,67R)-56-[(1R)-2-[(1S,3R,4R)-4-[2-[tert-butyl(diphenyl)silyl]oxyethoxy]-3-methoxy-cyclo [Hexyl]-1-methyl-ethyl-57,67-dihydroxy-55,58-dimethoxy-46,47,48,49,59,60-hexamethyl-77,78-dioxa-69-azatricyclohexatriaconta-35,37,39(59),40(60)-tetraene-61,62,63,64,65-pentone (4.7 g, 72% yield) was obtained as a yellow solid. ESI-MS (EI + , m / z): 1219.5 [M + Na] + .

[0451] Step 4: (36E,38E,40E,41E,47R,48S,49R,50R,52S,54S,56S,57S,58R,59R,68R)-57-[(1R)-2-[(1S,3R,4R)-4-[2-[tert-butyl(diphenyl)silyl]oxyethoxy]-3-methoxy-cyclohexyl]-1-methyl-ethyl]-68-hydroxy-56,58,59-trimethoxy-47,48,49,50,60,61-hexamethyl-77,78-dioxa-70-azatricyclohexatriaconta-36,38,40(60),41(61)-tetraene-62,63,64,65,66-pentone. (35E,37E,39E,40E,46R,47S,48R,49R,51S,53S,55S,56S,57R,58R,67R)-56-[(1R)-2-[(1S,3R,4R)-4-[2-[tert-butyl(diphenyl)silyl]oxyethoxy]-3-methoxy-cyclohexyl]-1-methyl-ethyl]-57,67-dihydroxy-55,58-dimethoxy-46,47,48,49 in toluene (40 mL) To a solution of 59,60-hexamethyl-77,78-dioxa-69-azatricyclohexatriaconta-35,37,39(59),40(60)-tetraene-61,62,63,64,65-pentone (2 g, 1.67 mmol) and 1,8-bis(dimethylamino)naphthalene (3.94 g, 18.39 mmol) was added methyl trifluoromethanesulfonate (2.19 g, 13.37 mmol) dropwise at room temperature under N. The mixture was then heated to 50 °C for 5 h, filtered, diluted with EA (60 mL), and washed with saturated NH4Cl(aq) (60 mL × 3), water (60 mL), and brine (60 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated.The residue was purified by silica gel chromatography (PE:EA=3:1) to give (36E,38E,40E,41E,47R,48S,49R,50R,52S,54S,56S,57S,58R,59R,68R)-57-[(1R)-2-[(1S,3R,4R)-4-[2-[tert-butyl(diphenyl)silyl]oxyethoxy]-3-methoxy-cyclohexane [xyl]-1-methyl-ethyl]-68-dihydroxy-56,58,59-trimethoxy-47,48,49,50,60,61-hexamethyl-77,78-dioxa-70-azatricyclohexatriaconta-36,38,40(60),41(61)-tetraene-62,63,64,65,66-pentone (700 mg, 35% yield) was obtained as a yellow solid. ESI-MS (EI). + , m / z): 1232.7 [M + Na] +

[0452] Step 5: (23E,25E,27E,28E,32R,33S,34R,35R,37S,39S,41S,42S,43R,44R,53R)-53-hydroxy-42-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-41,43,44-trimethoxy-32,33,34,35,45,46-hexamethyl-62,63-dioxa-54-azatricyclohexatriaconta-23,25,27(45),28(46)-tetraene-47,48,49,50,51-pentone (Intermediate V). (36E,38E,40E,41E,47R,48S,49R,50R,52S,54S,56S,57S,58R,59R,68R)-57-[(1R)-2-[(1S,3R,4R)-4-[2-[tert-butyl(diphenyl)silyl]oxyethoxy]-3-methoxy-cyclohexyl]-1-methyl-ethyl]-68-hydroxy-56 To a solution of 58,59-trimethoxy-47,48,49,50,60,61-hexamethyl-77,78-dioxa-70-azatricyclohexatriaconta-36,38,40(60),41(61)-tetraene-62,63,64,65,66-pentone (0.6 g, 0.495 mmol) was added pyridine HF (0.39 g, 4.96 mmol) at 0 °C. The mixture was stirred at 30 °C for 3 h, then quenched with saturated NaHCO solution (20 mL) and extracted with EA (30 mL) at 0 °C. The organic layer was washed with water (20 mL) and brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel chromatography (PE:acetone=3:1) to give intermediate V (430 mg, 89% yield) as a pale yellow solid. + , m / z): 994.7 [M + Na] + .

[0453] Synthesis of Intermediate VII

[0454] [ka]

[0455] Step 1: (27E,29E,31E,32E,35R,36S,37R,38R,40S,42S,44S,45S,46R,47R,57R)-46,57-dihydroxy-44,47-dimethoxy-45-[(1R)-2-[(1S,3R,4R)-3-methoxy-4-phenoxycarbothioyloxy-cyclohexyl]-1-methyl-ethyl]-35,36,37,38,48,49-hexamethyl-66,67-dioxa-58-azatricyclohexatriaconta-27,29,31(48),32(49)-tetraene-50,51,52,53,54-pentone. (22E,24E,26E,27E,29R,30S,31R,32R,34S,36S,38S,39S,40R,41R,50R)-40,50-dihydroxy-39-[(1R)-2-[(1S,3R,4R)-4-hydroxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-38,41-dimethoxy-29,30,31,32,42,43 in DCM (8 mL) To a solution of 1,3-hexamethyl-60,61-dioxa-51-azatricyclohexatriaconta-22,24,26(42),27(43)-tetraene-44,45,46,47,48-pentone (300 mg, 0.328 mmol) was added pyridine (208 mg, 2.63 mmol) and O-phenylcarbonochloridothioate (283 mg, 1.64 mmol) at 0 °C. The resulting solution was stirred at 0 °C for 2 h, then diluted with DCM, washed with NH Cl, water, and brine, dried over Na SO , filtered, and concentrated. The residue was purified by reverse-phase chromatography (0% to 100% CH CN in water) to give the title compound (150 mg, 44% yield) as a white solid. ESI-MS (EI + , m / z): 1072.3 [M + Na] + . 1H NMR(500MHz,CDCl3)δ 7.41(t,J=7.9Hz,2H),7.29(d,J=7.4Hz,1H),7.12(d,J=7.8Hz,2H),6.44-6.09(m,3H),5.96(d,J=10.4Hz,1H),5.61-5.38(m,2H),5.29(d,J=5.2Hz,1H),5.22-5.06(m,2H),4.79(s,1H),4.20(dd,J=16.6,6.0Hz,1H),3.93-3.52(m,4H),3.51-3.28(m,10H),3.14(s,3H),2.91-2.55(m,3H),2.25(dd,J=91.2,12.9Hz,4H),1.97(d,J=4.8Hz,2H),1.90-1.69(m,9H),1.60(t,J=22.2Hz,11H),1.54-1.38(m,7H),1.37-1.19(m,5H),1.11(ddd,J=25.6,13.0,7.6Hz,10H),1.01-0.84(m,10H)。

[0456] Step 2: (23E,25E,27E,28E,30R,31S,32R,33R,35S,37S,38S,39S,40R,41R,50R)-40,50-dihydroxy-38,41-dimethoxy-39-[(1R)-2-[(1S,3S)-3-methoxycyclohexyl]-1-methyl-ethyl]-30,31,32,33,42,43-hexamethyl-59,60-dioxa-51-azatricyclohexatriaconta-23,25,27(42),28(43)-tetraene-44,45,46,47,48-pentone. (27E,29E,31E,32E,35R,36S,37R,38R,40S,42S,44S,45S,46R,47R,57R)-46,57-dihydroxy-44,47-dimethoxy-45-[(1R)-2-[(1S,3R,4R)-3-methoxy-4-phenoxycarbothioyloxy-cyclohexyl]-1-methyl-ethyl]-35,36,37,38,48, To a solution of 49-hexamethyl-66,67-dioxa-58-azatricyclohexatriaconta-27,29,31(48),32(49)-tetraene-50,51,52,53,54-pentone (1.4 g, 1.33 mmol) was added triethylborane (157 mg, 1.60 mmol) and bis(trimethylsilyl)silyl-trimethyl-silane (994 mg, 4 mmol, 1 M in THF). The resulting solution was stirred at 100 °C for 1 h, then concentrated and purified by silica gel chromatography (0% to 50% EtOAc in PE) to give the title compound (0.6 g, 50% yield) as a pale yellow solid. ESI-MS (EI + , m / z): 920.0 [M + Na] + . 1H NMR(400MHz,CDCl3)δ 6.59-5.85(m,4H),5.68-5.06(m,4H),4.68(dd,J=48.1,31.4Hz,1H),4.49-3.99(m,2H),3.99-3.51(m,4H),3.52-3.27(m,7H),3.29-3(m,5H),2.88-2.53(m,3H),2.20(ddd,J=80.2,58.5,14.9Hz,6H),1.80(dd,J=34.0,5.5Hz,7H),1.63(d,J=16.1Hz,12H),1.52-1.19(m,10H),1.21-0.78(m,19H),0.70(dd,J=16.1,9.9Hz,2H)。

[0457] Step 3: (24E,26E,28E,29E,31R,32S,33R,34R,36S,38S,39S,40S,41R,42R,51R)-51-hydroxy-39,41,42-trimethoxy-40-[(1R)-2-[(1S,3S)-3-methoxycyclohexyl]-1-methyl-ethyl]-31,32,33,34,43,44-hexamethyl-59,60-dioxa-52-azatricyclohexatriaconta-24,26,28(43),29(44)-tetraene-45,46,47,48,49-pentone (Intermediate VI). (23E,25E,27E,28E,30R,31S,32R,33R,35S,37S,38S,39S,40R,41R,50R)-40,50-dihydroxy-38,41-dimethoxy-39-[(1R)-2-[(1S,3S)-3-methoxycyclohexyl]-1-methyl-ethyl]-30,31,32,33,42,43-hexamethyl-59,60-dioxa To a solution of sa-51-azatricyclohexatriaconta-23,25,27(42),28(43)-tetraene-44,45,46,47,48-pentone (200 mg, 0.222 mmol) was added N1,N1,N8,N8-tetramethylnaphthalene-1,8-diamine (668 mg, 3.12 mmol) and methyl trifluoromethanesulfonate (365 mg, 2.23 mmol). The resulting solution was stirred at 50 °C for 1 h, then cooled, filtered, and concentrated. The residue was purified by silica gel chromatography (45% EtOAc in PE) to give intermediate VI (50 mg, 12% yield) as a white solid. ESI-MS (EI + , m / z): 934.2 [M + Na] + . 1H NMR(400MHz,CDCl3)δ 6.54-5.81(m,4H),5.78-5.02(m,5H),4.52(dd,J=105.2,28.6Hz,1H),4.38-3.94(m,1H),3.93-3.53(m,4H),3.54-3.01(m,12 H),3.03-2.46(m,3H),2.45-1.88(m,6H),1.90-1.54(m,16H),1.54-1.19(m,9H),1.19-0.76(m,16H),0.70(d,J=11.0Hz,2H).

[0458] Compound synthesis examples Example 1: Synthesis of (24E,26E,28E,29E,35R,36S,37R,38R,40S,42S,44S,45R,46R,55R)-55-hydroxy-45,46-dimethoxy-44-[(1R)-2-[(1S,3S)-3-methoxycyclohexyl]-1-methyl-ethyl]-43-[2-(2-methoxyethoxy)ethoxy]-35,36,37,38,47,48-hexamethyl-63,64-dioxa-56-azatricyclohexatriaconta-24,26,28(47),29(48)-tetraene-49,50,51,52,53-pentone (I-1)

[0459] [ka]

[0460] To a solution of intermediate VII (150 mg, 0.164 mmol) and 2-(2-methoxyethoxy)ethanol (395 mg, 3.29 mmol) in THF (5 mL) was added HND-8 (25 mg) under Ar at 50 °C. The resulting solution was stirred at 50 °C for 2 h, then filtered and concentrated. The residue was purified by reverse-phase chromatography (85% CHCN in water) to give the title compound (I-1: 105 mg, 64% yield) as a white solid. ESI-MS (EI + , m / z): 1022.0 [M + Na] + . 1H NMR(500MHz,CDCl3)δ 6.51-5.83(m,4H),5.72-5.08(m,4H),4.41(ddd,J=101.7,68.7,23.6Hz,2H),4.01-3.03(m,22H), 2.93-2.50(m,5H),2.42-1.70(m,17H),1.52-1.21(m,16H),1.20-0.78(m,18H),0.77-0.65(m,1H).

[0461] Example 2: (3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-27-hydroxy-9,10-dimethoxy-3-((R)-1-((1S,3S)-3-methoxycyclohexyl)propan-2-yl)-21-(2-((2-methoxyethyl)sulfonyl)ethoxy)-6,8, Synthesis of 12,14,20,26-hexamethyl-9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-hexadecahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohentriacontin-1,5,11,28,29(4H,6H,31H)-pentaone (I-2):

[0462] [ka]

[0463] Step 1: 2-[2-[tert-Butyl(diphenyl)silyl]oxyethylsulfonyl]ethanol. To a solution of 2-(2-hydroxyethylsulfonyl)ethanol (5.01 g, 32.47 mmol) in pyridine (20 mL) was added tert-butyl-chloro-diphenyl-silane (2.22 g, 8.08 mmol) at 0 °C. The reaction was stirred at 15 °C for 3 h, then diluted with water (200 mL) and extracted with EtOAc (100 mL × 3). The combined organic layers were concentrated and purified by silica gel chromatography (EtOAc:PE = 1:2) to give 2-(2-(tert-butyl(diphenyl)silyloxy]oxyethylsulfonyl)ethanol (2.25 g, 71% yield) as a white solid. 1 H NMR(400MHz,CDCl3):δ 7.65-7.67(m,4H),7.42-7.47(m,6H),4.09-4.14(m,4H),3.44-3.46(m,2H),3.25-3.27(m,2H),2.57-2.60(m,1H),1.06(s,9H).

[0464] Step 2: tert-Butyl-[2-(2-methoxyethylsulfonyl)ethoxy]-diphenyl-silane. To a solution of 2-[2-[tert-butyl(diphenyl)silyl]oxyethylsulfonyl]ethanol (8.6 g, 21.91 mmol) and N1,N1,N8,N8-tetramethylnaphthalene-1,8-diamine (14.08 g, 65.72 mmol) in toluene (20 mL) was added methyl trifluoromethanesulfonate (10.78 g, 65.72 mmol) at 0° C. The mixture was stirred at 50° C. for 18 hours, then concentrated, treated with water (200 mL), and extracted with EtOAc (150 mL × 2). The combined organic layers were concentrated and purified by silica gel column chromatography (PE: EtOAc = 3:1) to give tert-butyl-[2-(2-methoxyethylsulfonyl)ethoxy]-diphenyl-silane (7.9 g, 89% yield) as a white solid. ESI-MS (EI + , m / z): 429.0 [M + Na] + . 1H NMR(400MHz,CDCl3):δ 7.67-7.69(m,4H),7.39-7.45(m,6H),4.07-4.10(m,2H),3.82-3.84(m ,2H),3.40-3.43(m,2H),3.37(s,3H),3.29-3.31(m,2H),1.06(s,9H).

[0465] Step 3: 2-(2-Methoxyethylsulfonyl)ethanol. To a solution of tert-butyl-[2-(2-methoxyethylsulfonyl)ethoxy]-diphenyl-silane (8.6 g, 21.15 mmol) in THF (10 mL) was added Py·HF (31.44 g, 317.26 mmol). The mixture was stirred at 20 °C for 18 h and then treated with EtOAc (100 mL). NaHCO3 (aq. 150 mL) was added, and the mixture was stirred at room temperature for 1 h, then filtered and washed with EtOAc (20 mL). The combined organic layers were concentrated and purified by reverse-phase chromatography (water) to give 2-(2-methoxyethylsulfonyl)ethanol (3.55 g, 99% yield) as a yellow oil. ESI-MS (EI + , m / z): 169.0 [M + H] + . 1 H NMR (400MHz, CDCl3): δ 4.09-4.13(m,2H),3.83-3.86(m,2H),3.40(s,3H),3.31-3.37(m,4H),2.68-2.71(m,1H).

[0466] Step 4: (24E,26E,28E,29E,35R,36S,37R,38R,40S,42S,44S,45R,46R,55R)-55-hydroxy-45,46-dimethoxy-44-[(1R)-2-[(1S,3S)-3-methoxycyclohexyl]-1-methyl-ethyl]-43-[2-(2-methoxyethylsulfonyl)ethoxy]-35,36,37,38,47,48-hexamethyl-65,66-dioxa-56-azatricyclohexatriaconta-24,26,28(47),29(48)-tetraene-49,50,51,52,53-pentone (I-2). To a solution of Intermediate VII (200 mg, 0.22 mmol) and 2-(2-methoxyethylsulfonyl)ethanol (369 mg, 2.19 mmol) in THF (3 mL) was added HND-8 (80 mg) at 50° C. The mixture was stirred at 50° C. for 6 h, then quenched with saturated NaHCO (aq) (20 mL) and extracted with EtOAc (30 mL). The organic layer was washed with water (20 mL) and brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by reverse-phase chromatography (0-100% CHCN in water) to give the title compound (I-2: 25 mg, 11% yield). ESI-MS (EI + , m / z): 1069.7 [M + Na] + . 1 H NMR (400 MHz, CDCl): δ 6.05-6.43(m,4H),5.10-5.59(m,4H),4.41-4.44(m,1H),3.53-3.90(m,7H),3 .24-3.46(m,15H),3.03-3.17(m,4H),2.89-2.95(m,1H),2.70-2.78(m,1H),2 .51-2.69(m,2H),2.17-2.34(m,4H),1.94-2.15(m,4H),1.54-1.89(m,25H),1 .22-1.53(m,12H),1.01-1.20(m,12H),0.84-0.96(m,8H),0.69-0.82(m,1H).

[0467] Example 3: (22E,24E,26E,27E,33R,34S,35R,36R,38S,40S,43S,44R,45R,55R)-43-[(1R)-2-[(1S,3R,4R)-4-difluoromethoxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-44,55-hydroxy-45-methoxy-42-[2-(2-methoxyethoxy)ethoxy]-33,34,35,36,46,47-hexamethyl-64,65-dioxa-56- Azatricyclohexatriaconta-22,24,26(46),27(47)-tetraene-48,49,50,51,52-pentone (I-3), (22E,24E,26E,27E,33R,34S,35R,36R,38S,40S,42S,43S,44R,45R,55R)-43-[(1R)-2-[(1S,3R,4R)-4-(difluoromethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-44, 55-hydroxy-45- Methoxy-42-[2-(2-methoxyethoxy)ethoxy]-33,34,35,36,46,47-hexamethyl-64,65-dioxa-56-azatricyclohexatriaconta-22,24,26(46),27(47)-tetraene-48,49,50,51,52-pentone (I-4), and (22E,24E,26E,27E,33R,34S,35R,36R,38S,40S,42R,43S,44R,45R,55R)-43-[(1R)- Synthesis of 2-[(1S,3R,4R)-4-(difluoromethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-44,55-dihydroxy-45-methoxy-42-[2-(2-methoxyethoxy)ethoxy]-33,34,35,36,46,47-hexamethyl-64,65-dioxa-56-azatricyclohexatriaconta-22,24,26(46),27(47)-tetraene-48,49,50,51,52-pentone (I-5)

[0468] [ka]

[0469] Step 1: (22E,24E,26E,27E,29R,30S,31R,32R,34S,36S,38S,39S,40R,41R,51R)-39-[(1R)-2-[(1S,3R,4R)-4-(difluoromethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-40,51-dihydroxy-38,41-dimethoxy-29,30,31,32,42,43-hexamethyl-60,61-dioxa-52-azatricyclohexatriaconta-22,24,26(42),27(43)-tetraene-44,45,46,47,48-pentone (Intermediate VIII). (22E,24E,26E,27E,29R,30S,31R,32R,34S,36S,38S,39S,40R,41R,50R)-40,50-dihydroxy-39-[(1R)-2-[(1S,3R,4R)-4-hydroxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-38,41-dimethoxy-29,30,31,32,42,43-hexamethyl- in DCM (15 mL) To a solution of 60,61-dioxa-51-azatricyclohexatriaconta-22,24,26(42),27(43)-tetraene-44,45,46,47,48-pentone (1.0 g, 1.09 mmol) in water (15 mL) was added potassium bifluoride (1.28 g, 16.41 mmol) and bromodifluoro(trimethylsilyl)methane (2.22 g, 10.94 mmol) at room temperature. The reaction was stirred at 25 °C for 18 h, then diluted with DCM, washed with saturated aqueous NH4Cl, water, and brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel chromatography (EtOAc:PE = 1:1.2) to give the title compound (110 mg, 10% yield) as a white solid. ESI-MS (EI + , m / z): 985.8 [M + Na] + . 1H NMR(400MHz,CDCl3)δ 6.72-5.83(m,5H),5.62(ddd,J=22.9,14.6,7.9Hz,1H),5.49-5.01(m,3H),4.67(s,1H),3.98-3.54(m,6H),3.52-3.05(m,15H),2.88-2.52(m,3H),2.41-1.68(m,16H),1.56-1.19(m,10H),1.17-0.86(m,17H),0.76(dd,J=24.3,12.0Hz,2H)。

[0470] Step 2: (22E,24E,26E,27E,33R,34S,35R,36R,38S,40S,43S,44R,45R,55R)-43-[(1R)-2-[(1S,3R,4R)-4-(difluoromethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-44,55-dihydroxy-45-methoxy-42-[2-(2-methoxyethoxy)ethoxy]-33,34,35,36,46,47-hexamethyl-64,65-dioxa-56-azatricyclohexatriaconta-22,24,26(46),27(47)-tetraene-48,49,50,51,52-pentone (CP-NAV-067-1410). (22E,24E,26E,27E,29R,30S,31R,32R,34S,36S,38S,39S,40R,41R,51R)-39-[(1R)-2-[(1S,3R,4R)-4-(difluoromethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-40,51-dihydroxy-38,41-dimethoxy-29,30,31,32,42,43-hexamethoxazole in THF (5 mL) To a solution of ethyl-60,61-dioxa-52-azatricyclohexatriaconta-22,24,26(42),27(43)-tetraene-44,45,46,47,48-pentone (200 mg, 0.21 mmol) and 2-(2-methoxyethoxy)ethanol (498 mg, 4.15 mmol) was added 4-methylbenzenesulfonic acid hydrate (197 mg, 1.04 mmol) under N at 0 °C. The reaction was stirred at this temperature for 2 h, then diluted with ice-cold aqueous NaHCO, extracted with EtOAc, washed with brine, dried over NaSO, filtered, and concentrated. The residue was purified by reverse-phase chromatography (76% CHCN in water) to give the title compound (I-3: 40 mg, 18% yield) as a white solid.

[0471] Step 3: (22E,24E,26E,27E,33R,34S,35R,36R,38S,40S,42S,43S,44R,45R,55R)-43-[(1R)-2-[(1S,3R,4R)-4-(difluoromethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-44,55-dihydroxy-45- Methoxy-42-[2-(2-methoxyethoxy)ethoxy]-33,34,35,36,46,47-hexamethyl-64,65-dioxa-56-azatricyclohexatriaconta-22,24,26(46),27(47)-tetraene-48,49,50,51,52-pentone (CP-NAV-067-1429-P1 ), and (22E,24E,26E,27E,33R,34S,35R,36R,38S,40S,42R,43S,44R,45R,55R)-43-[(1R)-2-[(1S,3R,4R)-4-(difluoromethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-44,55-dihydroxy-45-meth 100 mg of the mixture was separated by chiral HPLC and then purified by silica gel chromatography (PE:DCM:EtOAc:MeOH=3:3:1:0.3) to give the title compounds (I-4: 28 mg, 28% yield) and (I-5: 15 mg, 15% yield) as white solids.

[0472] Chiral separation methods: Column: CHIRALPAK IC Column size: 5.0 cm inner diameter x 25 cm length, 10 μm Sample solution: 1mg / mL in mobile phase Injection: 5mL Mobile phase: hexane / EtOH = 70 / 30 (V / V) Flow rate: 30mL / min Wavelength: UV254nm Temperature: 38℃ I-4:ESI-MS(EI + ,m / z):1073.7[M+Na] + 。 1 H NMR(500MHz,CDCl3)δ 6.60-6.05(m,4H),5.91(dd,J=41.4,11.1Hz,1H),5.58-5.07(m,4H),4.74(s,1H),4.19(dd,J=14.0,6.0Hz,1H),3.95-3.26(m,24H),3.12(dd,J=16.8,7.9Hz,1H),2.92-2.51(m,3H),2.40-1.86(m,8H),1.84-1.64(m,11H),1.54-1.16(m,10H),1.16-0.83(m,18H),0.78-0.65(m,1H)。 I-5:ESI-MS(EI + ,m / z):1073.7[M+Na] + 。 1 H NMR(500MHz,CDCl3)δ 6.64-5.88(m,5H),5.75-5.08(m,5H),4.28(s,1H),4.03-3.02(m,26H),2.98-1.90(m,9H),1.86-1.63(m,16H),1.50-1.17(m,6H),1.16-0.81(m,18H),0.78-0.61(m,1H)。

[0473] Example 4: (24E,26E,28E,29E,32R,33S,34R,35R,37S,39S,42S,44R,45R,55R)-42-[(1R)-2-[(1S,3R,4R)-4-(difluoromethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-41-(1,4-dioxan-2-ylmethoxy)-44,55-dihydroxy-45-methoxy-32,33,34,35,46,47-hexamethyl-66,67-dioxa-5 6-Azatricyclohexatriaconta-24,26,28(46),29(47)-tetraene-48,49,50,51,52-pentone (I-6), (24E,26E,28E,29E,32R,33S,34R,35R,37S,39S,41S,42S,44R,45R,55R)-42-[(1R)-2-[(1S,3R,4R)-4-(difluoromethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-41(1,4-dioxane- 2-ylmethoxy)-44,55-dihydroxy-45-methoxy-32,33,34,35,46,47-hexamethyl-66,67-dioxa-56-azatricyclohexatriaconta-24,26,28(46),29(47)-tetraene-48,49,50,51,52-pentone (I-9), and (24E,26E,28E,29E,32R,33S,34R,35R,37S,39S,41R,42S,44R,45R,55R)-42-[(1R )-2-[(1S,3R,4R)-4-(difluoromethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-41-(1,4-dioxan-2-ylmethoxy)-44,55-dihydroxy-45-methoxy-32,33,34,35,46,47-hexamethyl-66,67-dioxa-56-azatricyclohexatriaconta-24,26,28(46),29(47)-tetraene-48,49,50,51,52-pentone (I-10).

[0474] [ka]

[0475] Step 1: (24E,26E,28E,29E,32R,33S,34R,35R,37S,39S,42S,44R,45R,55R)-42-[(1R)-2-[(1S,3R,4R)-4-(difluoromethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-41-(1,4-dioxan-2-ylmethoxy)-44,55-dihydroxy-45-methoxy-32,33,34,35,46,47-hexamethyl-66,67-dioxa-56-azatricyclohexatriaconta-24,26,28(46),29(47)-tetraene-48,49,50,51,52-pentone (I-6). (22E,24E,26E,27E,29R,30S,31R,32R,34S,36S,38S,39S,40R,41R,51R)-39-[(1R)-2-[(1S,3R,4R)-4-(difluoromethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-40,51-dihydroxy-38,41-dimethoxy-29,30,31,32,42, in THF (10 mL). To a solution of 43-hexamethyl-60,61-dioxa-52-azatricyclohexatriaconta-22,24,26(42),27(43)-tetraene-44,45,46,47,48-pentone (0.1 g, 0.1 mmol, from Example 3) and 2-(oxetan-3-yloxy)ethanol (245 mg, 2.07 mmol) was added HND-8 (50 mg) under N at 50 °C. The reaction mixture was stirred at 50 °C for 20 h, then cooled, filtered, poured into saturated aqueous NaHCO (2 mL) at 0 °C, and extracted with EtOAc (20 mL). The organic layer was washed with water (20 mL) and brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by reverse-phase chromatography on silica gel (4:1 EtOAc:PE) eluting with 60% CH3CN in water to give the title compound (30 mg, 28% yield) as a white solid. ESI-MS (EI + , m / z): 1072.5 [M + Na] +. 1H NMR(400MHz,CDCl3)δ 6.58-5.92(m,5H),5.53-4.75(m,5H),4.27-4.09(m,2H),3.84-3.67(m,9H),3.63-3.54(m,2H),3.45-3.28(m,10H),3.25-3.07(m,3H),2 .84-2.55(m,3H),2.35-2.20(m,2H),2.13-1.86(m,6H),1.46-1.77(m,37H),1.43-1.17(m,14H),1.11-0.82(m,22H),0.79-0.69(m,1H).

[0476] Step 2: (24E,26E,28E,29E,32R,33S,34R,35R,37S,39S,41S,42S,44R,45R,55R)-42-[(1R)-2-[(1S,3R,4R)-4-(difluoromethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-41-(1,4-dioxan-2-ylmethoxy)-44,55-dihydroxy-45-methoxy-32,33,34,35,46,47-hexamethyl-66,67-dioxa-56-azatricyclohexatriaconta-24,26,28(46),29(47)-tetraene-48,49,50,51,52-pentone (I-9 ), and (24E,26E,28E,29E,32R,33S,34R,35R,37S,39S,41R,42S,44R,45R,55R)-42-[(1R)-2-[(1S,3R,4R)-4-(difluoromethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-41-(1,4-dioxan-2-ylmethoxy)-44,55-dihydroxy-45-methoxy-32,33,34,35,46,47-hexamethyl-66,67-dioxa-56-azatricyclohexatriaconta-24,26,28(46),29(47)-tetraene-48,49,50,51,52-pentone (I-10). The 115 mg mixture was separated by chiral HPLC and then purified by silica gel chromatography (PE:DCM:EtOAc:MeOH=3:3:1:0.3) to give the title compounds (I-9: 35 mg, 30% yield) and (I-10: 12 mg, 10% yield) as white solids.

[0477] Kirara separation method: カラム:CHIRALPAK IC カラムサイズ: 5.0cm inner diameter × 25cm length, 10μm Sample solution: 0.2 mg / mL in mobile phase Injection: 5mL Mobile phase: Hexane / EtOH=60 / 40 (V / V) Flow rate: 30mL / min Wavelength: UV266nm Temperature: 35℃ I-9:ESI-MS(EI + ,m / z):1072.5[M+Na] + . 1 H NMR (400 MHz, CDCl3) δ 6.59-6.10(m,4H),5.91(dd,J=28.2,10.4Hz,1H),5.58-5.06(m,4H),4.75(dd ,J=16.6,9.8Hz,1H),4.69-4.53(m,1H),4.17(d,J=5.7Hz,1H),3.91-3.54(m, 12H),3.48-3.01(m,13H),2.91-2.53(m,3H),2.38-1.81(m,7H),1.83-1.64(m ,9H),1.52-1.19(m,10H),1.16-0.81(m,18H),0.74(dd,J=24.3,12.0Hz,1H). I-10:ESI-MS(EI + ,m / z):1072.5[M+Na] + . 1 H NMR(400MHz, CDCl3)δ 6.67-5.84(m,5H),5.70-5.07(m,4H),4.37-4.07(m,3H),3.98(t,J=4.3Hz,1H),3.87-3.57(m,8H),3.56-3.0 5(m,13H),2.93-1.97(m,10H),1.94-1.64(m,15H),1.54-1.20(m,7H),1.18-0.83(m,18H),0.77-0.61(m,1H).

[0478] Example 5: Synthesis of (24E,26E,28E,29E,35R,36S,37R,38R,40S,42S,45S,47R,48R,57R)-47,57-dihydroxy-48-methoxy-44-[2-(2-methoxyethoxy)ethoxy]-45-[(1R)-2-[(1S,3R)-3-methoxy-4-(oxetan-3-yloxy)cyclohexyl]-1-methyl-ethyl]-35,36,37,38,49,50-hexamethyl-67,68-dioxa-58-azatricyclohexatriaconta-24,26,28(49),29(50)-tetraene-51,52,53,54,55-pentone (I-7).

[0479] [ka]

[0480] Step 1: [(37S,39R,41R)-4-[(2R)-(22E,24E,26E,27E,33R,34S,35R,36R,38S,40S,43S,44R,45R,54R)-44,54-dihydroxy-45-methoxy-42-[(2-(2 methoxyethoxy)ethoxy]-33,34,35,36,46,47-hexamethyl-48,49,50,51,52-pentaoxo-66, 67-Dioxa-56-azatricyclohexatriaconta-22,24,26(46),27(47)-tetra-43-yl]propyl]-41-methoxy-39-cyclohexyl]trifluoromethanesulfonate. (22E,24E,26E,27E,33R,34S,35R,36R,38S,40S,43S,44R,45R,54R)-44,54-dihydroxy-43- [(1R)-2-[(1S,3R,4R)-4-hydroxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-45-methoxy-42-[2-(2-methoxyethoxy)ethoxy]-33,34,35,36,46,47-hexamethyl-64,65-dioxa-55-azatricyclohexatriaconta-22,24,26(46),27(47)-tetraene-48,49,50,51,52 To a solution of -pentone (compound A prepared according to U.S. Pat. No. 10,980,784, 1 g, 1 mmol) and 2,6-dimethylpyridine (1.07 g, 10 mmol) was added trifluoromethanesulfonic anhydride (1.41 g, 4.99 mmol) (dissolved in 1 mL of DCM) dropwise under N at 0° C. The reaction was stirred at 0° C. for 0.5 h. The reaction mixture was used directly in the next step without further purification.

[0481] Step 2: (24E,26E,28E,29E,35R,36S,37R,38R,40S,42S,45S,47R,48R,57R)-47,57-dihydroxy-48-methoxy-44-[2-(2-methoxyethoxy)ethoxy]-45-[(1R)-2-[(1S,3R)-3-methoxy-4-(oxetan-3-yloxy)cyclohexyl]-1-methyl-ethyl]-35,36,37,38,49,50-hexamethyl-67,68-dioxa-58-azatricyclohexatriaconta-24,26,28(49),29(50)-tetraene-51,52,53,54,55-pentone (I-7). The reaction solution from Step 1 was cooled to 0 °C under N2, and DIPEA (1.29 g, 9.96 mmol) and oxetan-3-ol (0.74 g, 9.96 mmol) were added. The reaction was allowed to warm to room temperature and stirred for 20 h, then concentrated and purified by silica gel chromatography (80% EtOAc in PE) and reverse-phase chromatography (eluting with 60% CH3CN in water) to give the title compound (0.055 g, 5% yield) as a white solid. ESI-MS (EI + , m / z): 1079.9 [M + Na] + . 1H NMR(400MHz,CDCl3)δ 6.39-5.94(m,4H),5.54-5.12(m,4H),4.79-4.49(m,4H),4.27-3.98(m,2H),3.91-3.74(m,3H),3.63-3.52(m,9H),3. 50-3.12(m,13H),2.81-2.49(m,3H),2.26-1.97(m,4H),1.91-1.49(m,29H),1.53-1.12(m,12H),1.14-0.84(m,15H).

[0482] Example 6: Synthesis of (25E,27E,29E,30E,33R,34S,35R,36R,38S,40S,43S,45R,46R,56R)-43-[(1R)-2-[(1S,3R,4R)-4-(difluoromethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-42-(1,4-dioxan-2-ylmethoxy)-56-hydroxy-45,46-dimethoxy-33,34,35,36,47,48-hexamethyl-66,67-dioxa-57-azatricyclohexatriaconta-25,27,29(47),30(48)-tetraene-49,50,51,52,53-pentone (I-8).

[0483] [ka]

[0484] Step 1: 3-(2-benzyloxyethoxy)oxetane. To a solution of oxetan-3-ol (8 g, 108 mmol) and 2-bromoethoxymethylbenzene (34.84 g, 162 mmol) in DMF (20 mL) was added sodium hydride (5.18 g, 216 mmol) batchwise. The resulting solution was stirred at 0 °C for 2 h and at room temperature for 16 h. The reaction was then quenched with 50 mL of NH4Cl (saturated aqueous solution) and then extracted with EtOAc (50 mL × 2). The organic layers were combined and concentrated. The residue was purified by silica gel chromatography eluting with PE:EtOAc (8:1) to give 3-(2-benzyloxyethoxy)oxetane (12.4 g, 55% yield). 1 H NMR(400MHz,CDCl3)δ 7.32(s,4H),7.31-7.26(m,1H),4.72(dd,J=6.3,5.7Hz,2H),4.64-4.60(m, 2H),4.58(dd,J=8.6,3.0Hz,1H),4.54(s,2H),3.57(dt,J=5.6,2.7Hz,5H).

[0485] Step 2: 2-(oxetan-3-yloxy)ethanol. To a solution of 3-(2-benzyloxyethoxy)oxetane (8 g, 38.41 mmol) in MeOH (20 mL) was added Pd / C (4.09 g, 38.41 mmol, 10%) batchwise. The resulting solution was stirred at 60° C. for 16 h, then filtered and concentrated. The residue was purified by silica gel chromatography (PE:EtOAc=1:5) to give 2-(oxetan-3-yloxy)ethanol (2.96 g, 65% yield). 1 H NMR(400MHz, CDCl3)δ 4.79(dd,J=8.3,4.3Hz,2H),4.62(dt,J=10.1,4.9Hz,3H),3.75(d,J=3.9Hz,2H),3.54-3.45(m,2H),2.44(d,J=5.9Hz,1H).

[0486] Step 3: (22E,24E,26E,27E,29R,30S,31R,32R,34S,36S,38S,39S,40R,41R,51R)-39-[(1R)-2-[(1S,3R,4R)-4-(difluoromethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-40,51-dihydroxy-38,41-dimethoxy-29,30,31,32,42,43-hexyl samethyl-60,61-dioxa-52-azatricyclohexatriaconta-22,24,26(42),27(43)-tetraene-44,45,46,47,48-pentone: To a solution of rapamycin (1 g, 1.09 mmol) in DCM (15 mL) was added bromodifluoro(trimethylsilyl)methane (2.22 g, 10.94 mmol) dissolved in 30 mL of water at room temperature. The reaction was stirred at room temperature for 16 h and then poured into ice-cold saturated aqueous NaHCO3 solution (10 mL). The organic layer was washed with water (10 mL x 3) and brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel chromatography (EtOAc:PE = 1:1) to give the title compound (200 mg, 19% yield) as a white solid. ESI-MS (EI + , m / z): 986.5 [M + Na] + ,T=2.428 minutes.

[0487] Step 4: (23E,25E,27E,28E,30R,31S,32R,33R,35S,37S,39S,40S,41R,42R,52R)-40-[(1R)-2-[(1S,3R,4R)-4-(difluoromethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-52-hydroxy-39,41,42-trimethoxy-30 ,31,32,33,43,44-Hexamethyl-60,61-dioxa-53-azatricyclohexatriaconta-23,25,27(43),28(44)-tetraene-45,46,47,48,49-pentone: (22E,24E,26E,27E,29R,30S,31R,32R,34S,36S,38S,39S) in toluene (6 mL) ,40R,41R,51R)-39-[(1R)-2-[(1S,3R,4R)-4-(difluoromethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-40,51-dihydroxy-38,41-dimethoxy-29,30,31,32,42,43-hexamethyl-60,61-dioxa-52-azatricyclohexatriaconta To a solution of 22,24,26(42),27(43)-tetraene-44,45,46,47,48-pentone (300 mg, 0.31 mmol), N1,N1,N8,N8-tetramethylnaphthalene-1,8-diamine (867 mg, 4.04 mmol) and methyl trifluoromethanesulfonate (0.51 g, 3.11 mmol) were added dropwise at room temperature. The reaction was stirred at 50 °C for 2 h, then filtered, concentrated, and purified by silica gel chromatography (1:1.5 EtOAc:PE) and reverse-phase chromatography (85% CH3CN in water) to give the title compound (100 mg, 33% yield) as a white solid. ESI-MS (EI + , m / z): 1000.5 [M + Na] + .

[0488] Step 5: (25E,27E,29E,30E,33R,34S,35R,36R,38S,40S,43S,45R,46R,56R)-43-[(1R)-2-[(1S,3R,4R)-4-(difluoromethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-42-(1,4-dioxan-2-ylmethoxy)-56-hydroxy-45,46-dimethoxy-33,34,35,36,47,48-hexamethyl-66,67-dioxa-57-azatricyclohexatriaconta-25,27,29(47),30(48)-tetraene-49,50,51,52,53-pentone (I-8). (23E,25E,27E,28E,30R,31S,32R,33R,35S,37S,39S,40S,41R,42R,52R)-40-[(1R)-2-[(1S,3R,4R)-4-(difluoromethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-52-hydroxy-39,41,42-trimethoxy-30,31,32,33,4 To a solution of 3,44-hexamethyl-60,61-dioxa-53-azatricyclohexatriaconta-23,25,27(43),28(44)-tetraene-45,46,47,48,49-pentone (50 mg, 0.05 mmol) and 2-(oxetan-3-yloxy)ethanol (121 mg, 1.02 mmol) was added HND-8 (25 mg) under N at 50 °C. The reaction mixture was stirred at 50 °C for 20 h, then cooled, concentrated, and purified by silica gel chromatography (EtOAc:PE = 1:1) to give the title compound (7 mg, 13% yield) as a white solid. ESI-MS (EI + , m / z): 1086.6 [M + Na] + ,T=2.479 minutes. 1H NMR(400MHz,CDCl3)δ 6.51-5.88(m,4H),5.34(d,J=3.5Hz,4H),4.86-4.10(m,2H),3.94-3.51(m,8H),3.51-2.99(m,13H),2.85-2. 45(m,3H),2.46-1.97(m,6H),1.97-1.54(m,20H),1.55-1.21(m,11H),1.21-0.81(m,17H),0.83-0.65(m,2H).

[0489] Example 7: (3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-27-hydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propan-2-yl)-9,10-ditrimethoxy-6,8,12,14,20,26-hexamethyl-21-(2 Synthesis of (oxetan-3-yloxy)ethoxy)-9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-hexadecahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohentriacontin-1,5,11,28,29(4H,6H,31H)-pentaone (I-11):

[0490] [ka]

[0491] Step 1: (3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-3-((R)-1-((1S,3R,4R)-4-((tert-butyldimethylsilyl)oxy)-3-methoxycyclohexyl)propan-2-yl)-9,27-dihydroxy-10,21-dimethoxy To a solution of rapamycin (5 g, 5.47 mmol) in DMF (60 mL) was added imidazole (1.49 g, 21.88 mmol) at room temperature, followed immediately by tert-butyl-chloro-dimethyl-silane (2.47 g, 16.41 mmol). The mixture was stirred at 50° C. for 6 h and then poured into a mixture of ice-cold saturated aqueous NH4Cl (40 mL) and Et2O:petroleum ether (60 mL, 2:1). The organic layer was washed with saturated aqueous NH4Cl (20 mL), water and brine (20 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel chromatography (10% to 50% EtOAc in petroleum ether) to give the title compound (4 g, 71% yield) as a white solid. ESI-MS (EI + , m / z): 1050.5 [M + Na] + .

[0492] Step 2: (3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-3-((R)-1-((1S,3R,4R)-4-((tert-butyldimethylsilyl)oxy)-3-methoxycyclohexyl)propan-2-yl)-27-hydroxy-9,10,21-trimethoxysilane Ci-6,8,12,14,20,26-hexamethyl-9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-hexadecahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohentriacontin-1,5,11,28,29(4H,6H,31H)-pentaone. (3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-3-((R)-1-((1S,3R,4R)-4-((tert-butyldimethylsilyl)oxy)-3-methoxycyclohexyl)propan-2-yl)-9,27-dihydroxy-10,21-dimethoxy-6,8,12,14,20,26-hexamethyl-9,10,12,13,14,21,22,23,2 To a suspension of 4,25,26,27,32,33,34,34a-hexadecahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohentriacontin-1,5,11,28,29(4H,6H,31H)-pentaone (1 g, 0.97 mmol) and 1,8-bis(dimethylamino)naphthalene (2.5 g, 11.67 mmol), methyl trifluoromethanesulfonate (1.6 mL, 14.59 mmol) was added dropwise at room temperature under N2. The mixture was then heated to 50 °C for 6 h, then cooled, filtered, and concentrated. The residue was purified by silica gel chromatography to give the title compound (0.45 g, 0.43 mmol) as a white solid. ESI-MS (EI + , m / z): 1064.6 [M + Na] + .

[0493] Step 3: (3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-27-hydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propan-2-yl)-9,10,21-trimethoxy-6,8,12, 14,20,26-Hexamethyl-9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-hexadecahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohentriacontin-1,5,11,28,29(4H,6H,31H)-pentaone. (3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-3-((R)-1-((1S,3R,4R)-4-((tert-butyldimethylsilyl)oxy)-3-methoxycyclohexyl)propan-2-yl)-27-hydroxy-9,10,21-trimethoxy-6,8,12,14,20,26-hexamethyl-9, To a solution of 10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-hexadecahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohentriacontin-1,5,11,28,29(4H,6H,31H)-pentaone (0.4 g, 0.38 mmol) was added pyridine hydrofluoride (3.34 mL, 38.37 mmol) at 0 °C. The reaction was stirred at 45 °C for 5 h, then poured into a mixture of DCM and aqueous NaHCO, washed with water and brine, dried over NaSO, filtered, and concentrated. The residue was purified by silica gel chromatography to give the title compound (160 mg, 45% yield) as a white solid. ESI-MS (EI + , m / z): 949.9 [M + Na] + .

[0494] Step 4: (3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-27-hydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propan-2-yl)-9,10-ditrimethoxy-6,8,12,14,20,26-hexamethyl-21- (2-(oxetan-3-yloxy)ethoxy)-9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-hexadecahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohentriacontin-1,5,11,28,29(4H,6H,31H)-pentaone (I-11). (3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-27-hydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propan-2-yl)-9,10,21-trimethoxy-6,8,12,14,20,26-hexamethyl-9,10,12, To a solution of 13,14,21,22,23,24,25,26,27,32,33,34,34a-hexadecahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohentriacontin-1,5,11,28,29(4H,6H,31H)-pentaone (500 mg, 538.68 μmol), TFA (1.66 mL, 21.55 mmol) was added at −50° C. The mixture was stirred at the same temperature for 10 min, and then 2-(oxetan-3-yloxy)ethanol (1.91 g, 16.16 mmol) dissolved in DCM (0.2 mL) was added, and the mixture was stirred at −10° C. for 5 h. The reaction was diluted with a mixture of DCM and aqueous NaHCO, washed with water and brine, dried over NaSO, filtered, and concentrated. The residue was then purified by reverse-phase chromatography (70% CHCN in water) to give I-11. ESI-MS (EI + , m / z): 1036.3 [M + Na] + . 1H NMR(400MHz,CDCl3)δ 6.51-5.80(m,4H),5.75-5.03(m,4H),4.83-4.09(m,4H),3.99-3.53(m,7H),3.52-3.02(m,15H),3.01-2.44(m,5H),2.11(d dd,J=99.8,49.8,39.7Hz,7H),1.83-1.61(m,13H),1.52-1.20(m,10H),1.18-0.80(m,17H),0.69(dd,J=23.8,11.9Hz,1H).

[0495] Example 8: Synthesis of (28E,30E,32E,33E,39R,40S,41R,42R,44S,46S,49S,51R,52R,61R)-48-(1,4-dioxan-2-ylmethoxy)-51,61-dihydroxy-52-methoxy-49-[(1R)-2-[(1S,3R,4R)-3-4-(3-morpholinopropoxy)cyclohexyl]-1-methyl-ethyl]-39,40,41,42,53,54-hexamethyl-74,75-dioxa-63-azatricyclohexatriaconta-28,30,32(53),33(54)-tetraene-55,56,57,58,59-pentone (I-12):

[0496] [ka]

[0497] Step 1: 1,4-Dioxan-2-ylmethanol. A mixture of 2-(oxetan-3-yloxy)ethanol (7.77 g, 65.77 mmol) and HND-8 (2.33 g, 65.77 mmol) in THF (120 mL) was stirred at 50° C. for 3 h. The mixture was filtered and concentrated to give 1,4-dioxan-2-ylmethanol (6.97 g, 90% yield) as a colorless oil. 1 H NMR (400MHz, CDCl3) δ 3.87-3.39(m, 9H), 2.37-2.11(m, 1H).

[0498] Step 2: 3-Iodopropyl trifluoromethanesulfonate. A mixture of 3-iodopropan-1-ol (4 g, 21.51 mmol) and 2,6-lutidine (4.61 g, 43.01 mmol) in DCM (40 mL) was cooled to 0 °C under N2, and trifluoromethylsulfonyl trifluoromethanesulfonate (6.67 g, 23.66 mmol) was added dropwise. The resulting solution was stirred at 0 °C for 2 h, then quenched with 10% EtOAc in petroleum ether, passed through a short silica gel column, filtered, and concentrated to give 3-iodopropyl trifluoromethanesulfonate (6.72 g, 98% yield).

[0499] Step 3: (22E,24E,26E,27E,32R,33S,34R,35R,37S,39S,41S,42S,43R,44R,53R)-43,53-dihydroxy-42-[(1R)-2-[(1S,3R,4R)-4-(3-iodopropoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-41,44-dimethoxy-32,33,34,35,45,46-hexamethyl-62,63-dioxa-54-azatricyclohexatriaconta-22,24,26(45),27(46)-tetraene-47,48,49,50,51-pentone. A mixture of rapamycin (2 g, 2.19 mmol) and N-ethyl-N-isopropyl-propan-2-amine (4.24 g, 32.82 mmol) in toluene (40 mL) was stirred at 50 °C for 16 h. The mixture was poured into ice-cold saturated NaHCO (50 mL), washed twice with ice-water (60 mL) and brine (50 mL), then dried over anhydrous NaSO, filtered, and concentrated. The residue was purified by silica gel chromatography (petroleum ether:EA = 3:1) to give the title compound (1.45 g, 61% yield) as a pale yellow solid. ESI-MS (EI + , m / z): 1104.5 [M + Na] + .

[0500] Step 4: (26E,28E,30E,31E,36R,37S,38R,39R,41S,43S,45S,46S,47R,48R,57R)-47,57-dihydroxy-45,48-dimethoxy-46-[(1R)-2-[(1S,3R,4R)-3-methoxy-4-(3-morpholinopropoxy)cyclohexyl]-1-methyl-ethyl]-36,37,38,39,49,50-hexamethyl-68,69-dioxa-59-azatricyclohexatriaconta-26,28,30(49),31(50)-tetraene-51,52,53,54,55-pentone. (22E,24E,26E,27E,32R,33S,34R,35R,37S,39S,41S,42S,43R,44R,53R)-43,53-hydroxy-42-[(1R)-2-[(1S,3R,4R)-4-(3-iodopropoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-41,44-dimethoxy-32,33,3 A mixture of 4,35,45,46-hexamethyl-62,63-dioxa-54-azatricyclohexatriaconta-22,24,26(45),27(46)-tetraene-47,48,49,50,51-pentone (1.35 g, 1.25 mmol) and N-ethyl-N-isopropyl-propan-2-amine (0.48 g, 3.74 mmol) was stirred at room temperature for 16 hours. The reaction mixture was diluted with DCM and acidified to pH 5 with 1 N HCl. The organic phase was washed with H2O, passed through a phase separator, dried over anhydrous Na2SO4, and concentrated. The residue was purified by silica gel chromatography (EA: 5% NH3 / MeOH) to give the title compound (0.5 g, 37% yield) as a pale yellow solid. ESI-MS (EI+, m / z): 1042.0 [M+Na] + .

[0501] Step 5: (28E,30E,32E,33E,39R,40S,41R,42R,44S,46S,49S,51R,52R,61R)-48-(1,4-dioxan-2-ylmethoxy)-51,61-dihydroxy-52-methoxy-49-[(1R)-2-[(1S,3R,4R)-3-methoxy-4-(3-morpholinopropoxy)cyclohexyl]-1-methyl-ethyl]-39,40,41,42,53,54-hexamethyl-74,75-dioxa-63-azatricyclohexatriaconta-28,30,32(53),33(54)-tetraene-55,56,57,58,59-pentone (I-12). (26E,28E,30E,31E,36R,37S,38R,39R,41S,43S,45S,46S,47R,48R,57R)-47,57-dihydroxy-45,48-dimethoxy-46-[(1R)-2-[(1S,3R,4R)-3-methoxy-4-(3-morpholinopropoxy)cyclohexyl]-1-methyl-ethyl]-36,37,38,39,49,50-hexyl in DCM (16 mL) To a solution of dimethyl-68,69-dioxa-59-azatricyclohexatriaconta-26,28,30(49),31(50)-tetraene-51,52,53,54,55-pentone (0.4 g, 0.38 mmol) and 1,4-dioxan-2-ylmethanol (1.36 g, 11.52 mmol) was added 2,2,2-trifluoroacetic acid (1.75 g, 15.36 mmol) under N at 0 °C. The reaction mixture was stirred at -10 °C for 20 h, then washed with ice-saturated aqueous NaHCO (10 mL), water (10 mL × 3), and brine (10 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated. The reaction mixture was purified by reverse-phase chromatography eluting with 50% CHCN in water to give I-12 (156 mg, 34% yield). ESI-MS(EI + , m / z): 1150.7 [M + Na] + . 1H NMR(400MHz,CDCl3)δ 6.51-6.01(m,4H),5.47(d,J=43.7Hz,3H),5.33-5.10(m,2H),4.22(d,J=31.5Hz,2H),3.73(dd,J=48.2,40.9Hz,12H),3.39(d d,J=28.6,10.3Hz,10H),3.02(d,J=10.8Hz,3H),2.71(d,J=16.9Hz,9H),2.32(s,2H),2.12-1.37(m,31H),1.35-0.75(m,20H).

[0502] Example 9: (27E,29E,31E,32E,39R,40S,41R,42R,44S,46S,48S,49S,51R,52R,61R)-51,61-dihydroxy-52-methoxy-49-[(1R)-2-[(1S,3R,4R)-3-methoxy-4-(3-morpholinopropoxy)cyclohexyl]-1 -methyl-ethyl]-39,40,41,42,53,54-hexamethyl-48-[2-(oxetan-3-yloxy)ethoxy]-73,74-dioxa-63-azatricyclohexatriaconta-27,29,31(53),32(54)-tetraene-55,56,57,58,59-pentone (I-14), and (27E,29E,31E,32E,39R,40S,41R,42R,44S,46S,48R,49S,51R,52R,61R)-51,61-dihydroxy-52-methoxy-49-[(1R)-2-[(1S,3R,4R)-3-methoxy-4-(3-morpholinopropoxy)cyclohexyl]-1-methyl Synthesis of [3-ethyl]-39,40,41,42,53,54-hexamethyl-48-[2-(oxetan-3-yloxy)ethoxy]-73,74-dioxa-63-azatricyclohexatriaconta-27,29,31(53),32(54)-tetraene-55,56,57,58,59-pentone (I-13).

[0503] [ka]

[0504] Step 1: 3-(2-benzyloxyethoxy)oxetane. To a solution of oxetan-3-ol (10 g, 135 mmol) in DMF (160 mL) was added sodium hydride (3.24 g, 135 mmol) at 0 °C. The resulting solution was stirred at this temperature for 30 minutes, and then 2-bromoethoxymethylbenzene (43.55 g, 202.49 mmol) was added. The resulting solution was stirred at 0 °C for 2 hours and then at room temperature for 16 hours. The reaction was quenched by the addition of 800 mL of NH4Cl (saturated aqueous solution) and then extracted with 2 × 120 mL of ethyl acetate. The organic layers were combined and concentrated. The residue was purified by silica gel chromatography eluting with petroleum ether / EA (8:1) to give the title compound (16.4 g, 78.75 mmol) as a colorless liquid. ESI-MS (EI + , m / z): 231 [M + Na] + . 1 H NMR (400MHz, CDCl3) δ 7.41-7.23 (m, 6H), 4.79-4.70 (m, 2H), 4.68-4.52 (m, 6H), 3.62-3.53 (m, 4H).

[0505] Step 2: 2-(oxetan-3-yloxy)ethanol. To a solution of 3-(2-benzyloxyethoxy)oxetane (4 g, 19.21 mmol) in MeOH (20 mL) under N was added Pd / C (2.04 g, 19.21 mmol), and the resulting solution was stirred under H at 40 °C overnight, then filtered and concentrated. The residue was purified by silica gel chromatography eluting with petroleum ether:EA = 1:5 to give the title compound (2.1 g, 93% yield) as a colorless liquid. 1 H NMR(400MHz, CDCl3)δ 4.79(td,J=5.8,2.1Hz,2H),4.62(dt,J=10.2,4.9Hz,3H),3.80-3.69(m,2H),3.52-3.44(m,2H),2.36(s,1H).

[0506] Step 3: 3-Iodopropyl trifluoromethanesulfonate. To a mixture of 3-iodopropan-1-ol (4 g, 21.5 mmol) and 2,6-lutidine (4.61 g, 43 mmol) in DCM (40 mL) under N was added trifluoromethylsulfonyl trifluoromethanesulfonate (6.67 g, 23.66 mmol) dropwise at 0° C. under N. The resulting solution was stirred at 0° C. for 2 hours, then quenched with 10% EtOAc in petroleum ether and filtered through a short silica gel column. The filtrate was concentrated in vacuo to give the title compound (6.72 g, 98% yield) as a pale yellow liquid.

[0507] Step 4: (22E,24E,26E,27E,32R,33S,34R,35R,37S,39S,41S,42S,43R,44R,53R)-43,53-dihydroxy-42-[(1R)-2-[(1S,3R,4R)-4-(3-iodopropoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-41,44-dimethoxy-32,33,34,35,45,46-hexamethyl-62,63-dioxa-54-azatricyclohexatriaconta-22,24,26(45),27(46)-tetraene-47,48,49,50,51-pentone. A mixture of rapamycin (2 g, 2.19 mmol) and N-ethyl-N-isopropyl-propan-2-amine (5.72 mL, 32.82 mmol) in toluene (40 mL) was stirred at 50 °C for 16 h. The mixture was then poured into ice-cold saturated NaHCO (50 mL), washed with ice water (60 mL × 2), brine (50 mL), dried over anhydrous NaSO, filtered, and concentrated. The residue was purified by silica gel chromatography (petroleum ether:EA = 3:1) to give the title compound (1.45 g, 61% yield) as a pale yellow solid. ESI-MS (EI + , m / z): 1104.5 [M + Na] + .

[0508] Step 5: (26E,28E,30E,31E,36R,37S,38R,39R,41S,43S,45S,46S,47R,48R,57R)-47,57-dihydroxy-45,48-dimethoxy-46-[(1R)-2-[(1S,3R,4R)-3-methoxy-4-(3-morpholinopropoxy)cyclohexyl]-1-methyl-ethyl]-36,37,38,39,49,50-hexamethyl-68,69-dioxa-59-azatricyclohexatriaconta-26,28,30(49),31(50)-tetraene-51,52,53,54,55-pentone. (22E,24E,26E,27E,32R,33S,34R,35R,37S,39S,41S,42S,43R,44R,53R)-43,53-hydroxy-42-[(1R)-2-[(1S,3R,4R)-4-(3-iodopropoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-41,44-dimethoxy-32,33,34,3 A mixture of 5,45,46-hexamethyl-62,63-dioxa-54-azatricyclohexatriaconta-22,24,26(45),27(46)-tetraene-47,48,49,50,51-pentone (Intermediate II, 1.35 g, 1.25 mmol) and N-ethyl-N-isopropyl-propan-2-amine (0.65 mL, 3.74 mmol) was stirred at room temperature for 16 hours. The reaction mixture was diluted with DCM and acidified to pH = 5 with 1N aqueous HCl. The organic phase was washed with H2O, filtered through a phase separator, then dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified by silica gel chromatography (EA: 5% 7M NH3 in MeOH = 4:1) to give the title compound (498 mg, 37% yield) as a pale yellow solid. ESI-MS (EI + , m / z): 1042.0 [M + Na] + .

[0509] Step 6: (27E,29E,31E,32E,39R,40S,41R,42R,44S,46S,49S,51R,52R,61R)-51,61-dihydroxy-52-methoxy-49-[(1R)-2-[(1S,3R,4R)-3-methoxy-4-((3-morpholinopropoxy)cyclohexyl]-1-methyl-ethyl]-39,40,41,42,53,54-hexamethyl-48-[2-(oxetan-3-yloxy)ethoxy]-73,74- Dioxa-63-azatricyclohexatriaconta-27,29,31(53),32(54)-tetraene-55,56,57,58,59-pentone. (26E,28E,30E,31E,37R,38S,39R,40R,42S,44S,46S,47S,48R,49S,58R)-48-ethyl-49,58-dihydroxy-46-methoxy-47-[(1R)-2-[(1S,3R,4R)-3-methoxy-4-(3-morpholine)]] in DCM (30 mL). To a solution of [(trifluoropropyl)cyclohexyl]-1-methyl-ethyl]-37,38,39,40,50,51-hexamethyl-69,70-dioxa-60-azatricyclohexatriaconta-26,28,30(50),31(51)-tetraene-52,53,54,55,56-pentone (200 mg, 0.19 mmol) was added 2,2,2-trifluoroacetic acid (0.59 mL, 7.70 mmol) dropwise under N at −50° C. After addition, the reaction mixture was cooled to −50° C. The mixture was stirred at -10°C for 10 minutes, and then 2-(oxetan-3-yloxy)ethanol (682 mg, 5.77 mmol, dissolved in DCM) was added to the reaction mixture at the same temperature. The reaction was stirred at -10°C for 2 hours, then poured into saturated aqueous NaHCO3 solution (15 mL) at 0°C and extracted with DCM (20 mL). The organic layer was washed with water (50 mL) and brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by reverse-phase chromatography (eluted with 50% CH3CN in water) to give the title compound (40 mg, 18% yield) as a white solid. ESI-MS (EI + , m / z): 1126.69 [M+H] + . 1H NMR(400MHz,CDCl3)δ 6.39-5.98(m,4H),5.55-5.03(m,5H),4.78-4.43(m,4H),4.15(d,J=40.7Hz,2H),3.71(t,J=21.3Hz,6H),3.60-3.45(m,3H),3.46-3.14(m,10H),2.96(d,J=11.0Hz,3H),2.56(d,J=54.3Hz,8H),2.26(s,2H),2.17-2.03(m,2H),1.94(s,4H),1.80-1.32(m,15H),1.28-1.10(m,11H),1.06-0.56(m,19H)。

[0510] Step 7: (27E,29E,31E,32E,39R,40S,41R,42R,44S,46S,48S,49S,51R,52R,61R)-51,61-dihydroxy-52-methoxy-49-[(1R)-2-[(1S,3R,4R)-3-methoxy-4-(3-morpholinopropoxy)cyclohexyl]-1-methyl-ethyl]-39,40,41,42,53,54-hexamethyl-48-[2-(oxetan-3-yloxy)ethoxy]-73,74-dioxa-63-azatricyclohexatriaconta-27,29,31(53),32(54)-tetraene-55,56,57,58,59-pentone (I-14 ), and (27E,29E,31E,32E,39R,40S,41R,42R,44S,46S,48R,49S,51R,52R,61R)-51,61-dihydroxy-52-methoxy-49-[(1R)-2-[(1S,3R,4R)-3-methoxy-4-(3-morpholinopropoxy)cyclohexyl]-1- Methyl-ethyl]-39,40,41,42,53,54-hexamethyl-48-[2-(oxetan-3-yloxy)ethoxy]-73,74-dioxa-63-azatricyclohexatriaconta-27,29,31(53),32(54)-tetraene-55,56,57,58,59-pentone (I-13). 120 mg of (27E,29E,31E,32E,39R,40S,41R,42R,44S,46S,49S,51R,52R,61R)-51,61-dihydroxy-52-methoxy-49-[(1R)-2-[(1S,3R,4R)-3-methoxy-4-(3-morpholinopropoxy)cyclohexyl]-1-methyl-ethyl]-39,40,41,42,53,54-hexamethyl-48-[2-(oxetan-3-yloxy)ethoxy]-7 3,74-Dioxa-63-azatricyclohexatriaconta-27,29,31(53),32(54)-tetraene-55,56,57,58,59-pentone was separated by chiral preparative HPLC and then purified by silica gel chromatography (17% MeOH:DCM:EA:3:3:1 in petroleum ether) to give I-14 (7.2 mg, 6% yield) as a white solid and I-13 (14.8 mg, 12% yield) as a white solid.

[0511] Kirara separation method: カラム:CHIRALPAK IC カラムサイズ: 2.5cm inner diameter × 25cm length, 10μm Sample solution: 1.3 mg / mL in mobile phase Injection: 8mL Mobile phase: Hexane / EtOH=50 / 50 (V / V) Flow rate: 20mL / min Wavelength: UV254nm Temperature: 38℃ I-14:ESI-MS(EI + ,m / z):1126.8[M+H] + ;1148.9[M+Na] + . 1 H NMR(500MHz,CDCl3)δ 6.41-5.74(m,4H),5.51-5.01(m,4H),4.80-4.41(m,5H),4.10(d,J=5.4Hz,1H), 3.88-2.88(m,26H),2.72-2.13(m,12H),2.00-1.27(m,26H),1.10-0.59(m,20H). I-13:ESI-MS(EI + ,m / z):1126.8[M+H] + ;1148.9[M+Na] + . 1 H NMR(500MHz,CDCl3)δ 6.41-5.80(m,4H),5.63-5.01(m,4H),4.76-4.45(m,5H),4.25-3.92(m,2H),3.87-2.88(m,25H),2.71-2.04(m ,12H),1.91(d,J=28.3Hz,5H),1.62(ddt,J=39.3,32.9,10.5Hz,14H),1.47-1.29(m,7H),1.08-0.55(m,20H).

[0512] Example 10: (28E,30E,32E,33E,40R,41S,42R,43R,45S,47S,50S,52R,53R,62R)-52,62-dihydroxy-53-methoxy-50-[(1R)-2-[(1S,3R,4R)-3-methoxy-4-[3-(4-methylpiperazin-1-yl)propoxy]cyclohexyl Synthesis of ]-1-methyl-ethyl]-40,41,42,43,54,55-hexamethyl-49-[2-(oxetan-3-yloxy)ethoxy]-74,75-dioxa-65-azatricyclohexatriaconta-28,30,32(54),33(55)-tetraene-56,57,58,59,60-pentone (I-15):

[0513] [ka]

[0514] Step 1: (27E,29E,31E,32E,37R,38S,39R,40R,42S,44S,46S,47S,48R,49R,58R)-48,58-dihydroxy-46,49-dimethoxy-47-[(1R)-2-[(1S,3R,4R)-3-methoxy-4-[3-(4-methylpiperazin-1-yl)propoxy]cyclohexyl]-1-methyl-ethyl]-37,38,39,40,50,51-hexamethyl-69,70-dioxa-61-azatricyclohexatriaconta-27,29,31(50),32(51)-tetraene-52,53,54,55,56-pentone. (22E,24E,26E,27E,32R,33S,34R,35R,37S,39S,41S,42S,43R,44R,53R)-43,53-hydroxy-42-[(1R)-2-[(1S,3R,4R)-4-(3-iodopropoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-41,44-dimethoxy-32,33,34,35,45,46-hexamethyl-62,63-dioxa-54 in DCM (30 mL) A mixture of -azatricyclohexatriaconta-22,24,26(45),27(46)-tetraene-47,48,49,50,51-pentone (Intermediate II, 1.45 g, 1.34 mmol), 1-methylpiperazine (0.16 g, 1.61 mmol), and N-ethyl-N-isopropyl-propan-2-amine (0.52 g, 4.02 mmol) was stirred at room temperature for 16 h, then diluted with DCM and acidified to pH 5 with 1 N HCl. The organic phase was washed with HO, passed through a phase separator, dried over anhydrous NaSO, and concentrated. The residue was purified by silica gel chromatography (EA: 5% NH / MeOH) to give the title compound (0.96 g, 68% yield) as a pale yellow solid. 1H NMR(400MHz,CDCl3):δ 5.95-6.39(m,4H),5.16-5.55(m,4H),4.10-4.22(m,2H),3.55-3.87(m,6H),3.30-3.43(m,8H),2.98-3.17(m,6H),2.67-2.86(m,9H),2.50-2.64(m,3H),2.46(S,2H),2.27-2.35(m,2H),1.95-2.05(m,7H),1.79-1.86(m,3H),1.74-1.76(m,3H),1.58-1.71(m,8H),1.46-1.54(m,3H),1.31-1.35(m,2H),0.86-1.35(m,23H)。

[0515] Step 2: (28E,30E,32E,33E,40R,41S,42R,43R,45S,47S,50S,52R,53R,62R)-52,62-dihydroxy-53-methoxy-50-[(1R)-2-[(1S,3R,4R)-3-methoxy-4-[3-(4-methylpiperazin-1-yl)propoxy]cyclohexyl] [yl]-1-methyl-ethyl]-40,41,42,43,54,55-hexamethyl-49-[2-(oxetan-3-yloxy)ethoxy]-74,75-dioxa-65-azatricyclohexatriaconta-28,30,32(54),33(55)-tetraene-56,57,58,59,60-pentone (I-15). (27E,29E,31E,32E,37R,38S,39R,40R,42S,44S,46S,47S,48R,49R,58R)-48,58-dihydroxy-46,49-dimethoxy-47-[(1R)-2-[(1S,3R,4R)-3-methoxy-4-[3-(4-methylpiperazin-1-yl)propoxy]cyclohexyl]-1-methyl- To a solution of [ethyl]-37,38,39,40,50,51-hexamethyl-69,70-dioxa-61-azatricyclohexatriaconta-27,29,31(50),32(51)-tetraene-52,53,54,55,56-pentone (0.78 g, 0.74 mmol), 2,2,2-trifluoroacetic acid (3.38 g, 29.65 mmol) was added dropwise under N at −40° C. After the addition, the reaction mixture was stirred at −40° C. for 10 min, and then 2-(oxetan-3-yloxy)ethanol (2.63 g, 22.23 mmol, in DCM) was added to the reaction mixture at the same temperature. The reaction was stirred at -20 °C for 2 h, then poured into saturated aqueous NaHCO (25 mL) at 0 °C and extracted with DCM (25 mL). The organic layer was washed with water (25 mL) and brine (25 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo. The residue was purified by reverse-phase chromatography eluting with 50% CHCN in 0.01% HCOOH in water to afford I-15 (120 mg, 14% yield) as a white solid. ESI-MS (EI + , m / z): 1140.8 [M + Na] + . 1H NMR(400MHz,CDCl3):δ 5.97-6.36(m,4H),5.15-5.49(m,4H),4.57-4.79(m,5H),4.04-4.27(m,2H),3.73-3.86(m,2H),3.54-3.64(m,3H),3.33-3.52(m,11H),2.64-3.09(m,15H),2.48-2.63(m,4H),2.28-2.35(m,2H),1.86-2.11(m,8H),1.61-1.79(m,11H),1.14-1.56(m,12H),0.82-1.09(m,18H)。

[0516] Example 11: [(40S,42R,44R)-4-[(2R)-2-[(27E,29E,31E,32E,36R,37S,38R,39R,41S,43S,45S,46S,47R,48R,58R)-47,58-dihydroxy-45,48-dimethoxy-36,37,38,39,49,50-hexamethyl-51,52,53,54,55-pentaoxo-71,72-dioxa-60-azatricyclohexatriaconta-27,29,31(49),32(50)-tetraene-4 6-yl]propyl]-44-methoxy-42-cyclohexyl]N-methyl-N-(2-morpholinoethyl)carbamate (I-16), [(43S,45R,47R)-4-[(2R)-2-[(28E,30E,32E,33E,39R,40S,41R,42R,44S,46S,48S,49S,51R,52R,62R)-51,62-dihydroxy-52-methoxy-39,40,41,42,53,54-hexamethyl-48-[2-(oxetan-3-yloxy)ethoxy]-5 5,56,57,58,59-pentaoxo-76,77-dioxa-64-azatricyclohexatriaconta-28,30,32(53),33(54)-tetraen-49-yl]propyl]-47-methoxy-45-cyclohexyl]N-methyl-N-(2-morpholinoethyl)carbamate (I-18), and [(43S,45R,47R)-4-[(2R)-2-[(28E,30E,32E,33E,39R,40S,41R,42R,44S,46S,48R,49S,51R,52 Synthesis of (R,62R)-51,62-dihydroxy-52-methoxy-39,40,41,42,53,54-hexamethyl-48-[2-(oxetan-3-yloxy)ethoxy]-55,56,57,58,59-pentaoxo-76,77-dioxa-64-azatricyclohexatriaconta-28,30,32(53),33(54)-tetraen-49-yl]propyl]-47-methoxy-45-cyclohexyl]N-methyl-N-(2-morpholinoethyl)carbamate (I-17)

[0517] [ka]

[0518] Step 1: tert-Butyl N-(2-morpholinoethyl)carbamate. To a solution of 2-morpholinoethanamine (10 g, 76.81 mmol) in DCM (5 mL) was added triethylamine (5.35 mL, 38.41 mmol) and tert-butoxycarbonyl tert-butyl carbonate (18.44 g, 84.5 mmol) at 0 °C, and the resulting solution was stirred at 25 °C overnight. The reaction mixture was diluted with 200 mL of dichloromethane and then washed with 30 mL of 10% sodium bicarbonate and 30 mL of brine. The organic layer was dried over magnesium sulfate, filtered, and concentrated to give the title compound (17 g, 96% yield) as an off-white solid. ESI-MS (EI + , m / z): 231.3 [M + H] + . 1 H NMR(400MHz, CDCl3)δ 3.78-3.62(m,4H),3.24(d,J=5.5Hz,2H),2.45(dd,J=8.0,3.9Hz,6H),1.49-1.42(m,9H).

[0519] Step 2: tert-Butyl N-methyl-N-(2-morpholinoethyl)carbamate. tert-Butyl N-(2-morpholinoethyl)carbamate (18 g, 78.16 mmol) was dissolved in DMF (240 mL) cooled to 0 °C, and NaH (9.38 g, 234.47 mmol, 60% purity) was added. The reaction was stirred at room temperature for 20 minutes, then cooled to 0 °C. Iodomethane (12.2 g, 85.97 mmol) was added, and the mixture was stirred for an additional 3 hours. The reaction was then diluted with ethyl acetate (500 mL) and washed sequentially with saturated aqueous ammonium chloride (300 mL) and brine (300 mL × 5). The organic layer was dried over sodium sulfate, filtered, and evaporated under reduced pressure to give the title compound (14 g, 73% yield) as a white solid. ESI-MS (EI + , m / z): 245.3 [M + H] + . 1H NMR(400MHz, CDCl3)δ 3.74-3.64(m,4H),3.34(s,2H),2.93-2.81(m,3H),2.48(d,J=4.8Hz,6H),1.46(s,10H).

[0520] Step 3: N-methyl-2-morpholino-ethanamine. To hydrochloric acid (4 M, 143.25 mL) at 0 °C, tert-butyl N-methyl-N-(2-morpholinoethyl)carbamate (14 g, 57.3 mmol) was added, and the mixture was stirred at room temperature for 50 minutes. The reaction mixture was concentrated, and the residue was treated with NH3 (7 M, 81.86 mL), stirred for 1 hour, and then concentrated. The residue was purified by silica gel chromatography (DCM:MeOH:TEA = 90:10:0.1) to give the title compound (7.4 g, 90% yield) as a yellow solid. ESI-MS (EI + , m / z): 145.1 [M + H] + . 1 H NMR (400MHz, DMSO-d6) δ 9.03 (s, 2H), 3.80 (s, 4H), 3.26 (dd, J=44.9, 20.4Hz, 8H), 2.63 (s, 3H).

[0521] Step 4: [(43S,45R,47R)-4-[(2R)-2-[(30E,32E,34E,35E,39R,40S,41R,42R,44S,46S,48S,49S,50R,51R,61R)-61-hydroxy-48,51-dimethoxy-39,40,41,42,52,53-hexamethyl-54,55,56,57,58-pentaoxo-50-trimethylsilyloxy-73,74-dioxa-63-azatricyclohexatriaconta-30,32,34(52),35(53)-tetraen-49-yl]propyl]-47-methoxy-45-cyclohexyl]N-methyl-N-(2-morpholinoethyl)carbamate. (25E,27E,29E,30E,32R,33S,34R,35R,37S,39S,41S,42S,43R,44R,53R)-53-hydroxy-42-[(1R)-2-[(1S,3R,4R)-4-hydroxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-41,44-dimethoxy-32,33,34,35,45,46-hexamethyl-43-trimethylsilyloxy] To a solution of si-62,63-dioxa-54-azatricyclohexatriaconta-25,27,29(45),30(46)-tetraene-47,48,49,50,51-pentone (0.5 g, 0.507 mmol) and pyridine (160.4 mg, 2.03 mmol, 164 μL) at 0 °C under argon, triphosgene (150.43 mg, 0.507 mmol) in THF (20 mL) was added dropwise via syringe. The reaction mixture was stirred at 0° C. for 1 h, then triethylamine (0.41 g, 4.06 mmol) and N-methyl-2-morpholino-ethanamine (1.46 g, 10.14 mmol) were added, and the resulting solution was stirred at 0° C. for an additional 1 h, diluted with DCM, washed with aqueous NH4Cl, water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel chromatography (8% MeOH in DCM) to give the title compound (386 mg, 66% yield) as a pale yellow solid. ESI-MS (EI + , m / z): 1156.4 [M + H] + . 1H NMR(400MHz,CDCl3)δ 6.57-5.93(m,4H),5.73-5.47(m,1H),5.27-4.98(m,2H),4.72(s,1H),4.56(s,1H),4.36-3.54(m,12H),3.54-3.05(m,12H),2.93(s,4H),2.40(dt,J=34.4,23.8Hz,11H),2.04(s,5H),1.88-1.52(m,12H),1.52-1.17(m,10H),1.20-0.73(m,17H),0.10--0.14(m,9H)。

[0522] Step 5: [(40S,42R,44R)-4-[(2R)-2-[(27E,29E,31E,32E,36R,37S,38R,39R,41S,43S,45S,46S,47R,48R,58R)-47,58-dihydroxy-45,48-dimethoxy-36,37,38,39,49,50-hexamethyl-51,52,53,54,55-pentaoxo-71,72-dioxa-60-azatricyclohexatriaconta-27,29,31(49),32(50)-tetraen-46-yl]propyl]-44-methoxy-42-cyclohexyl]N-methyl-N-(2-morpholinoethyl)carbamate. [(43S,45R,47R)-4-[(2R)-2-[(30E,32E,34E,35E,39R,40S,41R,42R,44S,46S,48S,49S,50R,51R,61R)-61-hydroxy-48,51-dimethoxy-39,40,41,42,52,53-hexamethyl-54,55,56,57,58-pentaoxo-50-trimethylsilyloxy-73, To a solution of 74-dioxa-63-azatricyclohexatriaconta-30,32,34(52),35(53)-tetraen-49-yl]propyl]-47-methoxy-45-cyclohexyl]N-methyl-N-(2-morpholinoethyl)carbamate (1.8 g, 1.56 mmol) in acetone (5 mL) and water (5 mL) was added 0.5 N sulfuric acid (4.67 mL) at 0 °C. The resulting solution was stirred at 0 °C for 2 h and then poured into a mixture of 100 mL of EtOAc and 100 mL of saturated aqueous NaHCO3. The organic layer was washed with water and brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel chromatography (5% MeOH in DCM) to give the title compound (1.4 g, 83% yield) as a pale yellow solid. 1H NMR(400MHz,CDCl3)δ 6.47-5.84(m,4H),5.60-5.05(m,4H),4.77(s,1H),4.55(s,1H),4.34-4.10(m,1H),3.92-3.52(m,7H),3.52-3.23(m,10H),3.13(d,J=2.7Hz,4H),2.92(s,3H),2.78-2.39(m,8H),2.40-2.00(m,5H),2.03-1.53(m,18H),1.53-1.11(m,12H),1.11-0.87(m,13H),0.83(d,J=6.5Hz,2H)。

[0523] Step 6: [(43S,45R,47R)-4-[(2R)-2-[(28E,30E,32E,33E,39R,40S,41R,42R,44S,46S,49S,51R,52R,62R)-51,62-dihydroxy-52-methoxy-39,40,41,42,53,54-hexamethyl-48-[2-(oxetan-3-yl)methyl] Oxy)ethoxy]-55,56,57,58,59-pentaoxo-76,77-dioxa-64-azatricyclohexatriaconta-28,30,32(53),33(54)-tetraen-49-yl]propyl]-47-methoxy-45-cyclohexyl]N-methyl-N-(2-morpholinoethyl)carbamate (I-16). [(40S,42R,44R)-4-[(2R)-2-[(27E,29E,31E,32E,36R,37S,38R,39R,41S,43S,45S,46S,47R,48R,58R)-47,58-hydroxy-45,48-dimethoxy-36,37,38,39,49,50-hexamethyl-51,52,53,54,55-pentaoxo-71,72-dioxa-60-azatricyclohexatriaconta-27,29,31(49)] in DCM (5 mL) under nitrogen. To a solution of [32(50)-tetraen-46-yl]propyl]-44-methoxy-42-cyclohexyl]N-methyl-N-(2-morpholinoethyl)carbamate (0.2 g, 0.18 mmol) was added TFA (426 μL, 5.53 mmol) at −10° C., followed by 2-(oxetan-3-yloxy)ethanol (0.436 g, 3.69 mmol). The mixture was stirred at −10° C. for 2 h, then washed with aqueous NaHCO, water, and brine, filtered, and concentrated. The residue was purified by reverse-phase chromatography (65% CHCN in water) to give I-16 (63 mg, 29% yield) as a white solid. ESI-MS (EI + , m / z): 1170.8 [M + H] + . 1H NMR(400MHz,CDCl3)δ 6.46-6.01(m,4H),5.56-5.15(m,4H),4.75(s,2H),4.60(s,3H),4.18(s,2H),3.72(s,6H),3.64-3.03 (m,13H),2.94(s,3H),2.80-2.28(m,9H),2.13-1.87(m,4H),1.84-1.38(m,24H),1.85-0.75(m,25H).

[0524] Step 7: [(43S,45R,47R)-4-[(2R)-2-[(28E,30E,32E,33E,39R,40S,41R,42R,44S,46S,48S,49S,51R,52R,62R)-51,62-dihydroxy-52-methoxy-39,40,41,42,53,54-hexamethyl-48-[2-(oxetane-3 -yloxy)ethoxy]-55,56,57,58,59-pentaoxo-76,77-dioxa-64-azatricyclohexatriaconta-28,30,32(53),33(54)-tetraen-49-yl]propyl]-47-methoxy-45-cyclohexyl]N-methyl-N-(2-morpholinoethyl)carbamate (I-18 ), and [(43S,45R,47R)-4-[(2R)-2-[(28E,30E,32E,33E,39R,40S,41R,42R,44S,46S,48R,49S,51R,52R,62R)-51,62-dihydroxy-52-methoxy-39,40,41,42,53,54-hexamethyl-48-[2-(oxetan-3-yl)methyl]

[0047] 120 mg of I-16 was separated by chiral preparative HPLC to give I-18 (27.5 mg, 23% yield) as a white solid and I-17 (16.1 mg, 13% yield) as a white solid.

[0525] Chiral analysis methods: カラム:CHIRALPAK IC(IC00CE-UF123) カラムサイズ: 0.46cm inner diameter × 25cm length Injection: 20 μL Mobile phase: EtOH = 100% Flow rate: 1.0mL / min Wavelength: UV254nm Temperature: 35℃ I-18:ESI-MS(EI + ,m / z):1170.8[M+H] + . 1 H NMR(400MHz, CDCl3)δ 6.40-5.78(m,4H),5.62-5.03(m,4H),4.76-4.39(m,6H),4.12(d,J=5.9Hz,1H),3.88-3.59(m,6H),3.56-3.00(m,17H) ,2.86(s,3H),2.79-2.18(m,11H),2.15-1.81(m,5H),1.59(t,J=15.2Hz,13H),1.49-1.15(m,11H),1.10-0.66(m,18H). I-17:ESI-MS(EI + ,m / z):1170.7[M+H] + . 1 H NMR(400MHz, CDCl3)δ 6.49-5.81(m,4H),5.64-5.06(m,4H),4.66(d,J=65.7Hz,3H),4.23(d,J=26.2Hz,2H),3.94-3 .03(m,28H),2.98-2.22(m,15H),2.21-1.69(m,11H),1.54-1.18(m,13H),1.15-0.69(m,19H).

[0526] Example 12: [(42S,44R,46R)-4-[(2R)-2-[(27E,29E,31E,32E,38R,39S,40R,41R,43S,45S,48S,50R,51R,61R)-50,61-dihydroxy-51-methoxy-38,39,40,41,52,53-hexamethyl-47-[2-(oxetan-3-yloxy)ethoxy]-54,55,56,57,58-pentaoxo-75,76-dioxa-64-azatricyclohexatriaconta-27,29,31 (52),32(53)-tetraen-48-yl]propyl]-46-methoxy-44-cyclohexyl]N-(2-morpholinoethyl)carbamate (I-19), [(42S,44R,46R)-4-[(2R)-2-[(27E,29E,31E,32E,38R,39S,40R,41R,43S,45S,47S,48S,50R,51R,61R)-50,61-dihydroxy-51-methoxy-38,39,40,41,52,53-hexamethyl-47-[2-(oxetane-3 -yloxy)ethoxy]-54,55,56,57,58-pentaoxo-75,76-dioxa-64-azatricyclohexatriaconta-27,29,31(52),32(53)-tetraen-48-yl]propyl]-46-methoxy-44-cyclohexyl]N-(2-morpholinoethyl)carbamate (I-21), and [(42S,44R,46R)-4-[(2R)-2-[(27E,29E,31E,32E,38R,39S,40R,41R,43S,45S,47R,4 8S,50R,51R,61R)-50,61-dihydroxy-51-methoxy-38,39,40,41,52,53-hexamethyl-47-[2-(oxetan-3-yloxy)ethoxy]-54,55,56,57,58-pentaoxo-75,76-dioxa-64-azatricyclohexatriaconta-27,29,31(52),32(53)-tetraen-48-yl]propyl]-46-methoxy-44-cyclohexyl]N-(2-morpholinoethyl)carbamate (I-20)

[0527] [ka]

[0528] Step 1: (25E,27E,29E,30E,32R,33S,34R,35R,37S,39S,41S,42S,43R,44R,53R)-53-hydroxy-42-[(1R)-2-[(1S,3R,4R)-4-hydroxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-41,44-dimethoxy-32,33,34,35,45,46-hexamethyl-43-trimethylsilyloxy-62,63-dioxa-54-azatricyclohexatriaconta-25,27,29(45),30(46)-tetraene-47,48,49,50,51-pentone. To a solution of rapamycin (5.5 g, 6.02 mmol) and imidazole (3.2 g, 48 mmol) in EtOAc (35 mL) was added TMSCl (5.2 g, 48 mmol) dropwise at 0 °C. After the addition, the resulting solution was stirred at room temperature for 1 h, and then 0.5 N H2SO4 solution (24 mL) was added at 0 °C. After the addition, the resulting solution was stirred at 0 °C for 1.5 h, and then diluted with EtOAc (100 mL) and brine (50 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified by silica gel chromatography to give the title compound (4.33 g, 73% for two steps) as a white solid. ESI-MS (EI + , m / z): 1008.2 [M + Na] + . 1 H-NMR(400MHz,CDCl3)δ 6.50-5.80(m,4H),5.61(ddd,J=23.0,14.1,7.6Hz,1H),5.37-5.19(m,2H),5.07(ddd,J=11.3,9. 1,5.2Hz,1H),4.71(d,J=1.4Hz,1H),3.89-3.56(m,4H),3.50-3.30(m,6H),3.29-3.18(m,3H),3. 18-3.04(m,3H),2.97-2.86(m,1H),2.84-2.45(m,3H),2.43-2.05(m,4H),1.97(dd,J=10.0,5.4H z,2H),1.86-1.50(m,19H),1.49-0.81(m,23H),0.68(dd,J=23.6,11.9Hz,1H),0.05-0.07(m,9H).

[0529] Step 2: [(42S,44R,46R)-4-[(2R)-2-[(29E,31E,33E,34E,38R,39S,40R,41R,43S,45S,47S,48S,49R,50R,60R)-60-hydroxy-47,50-dimethoxy-38,39,40,41,51,52-hexamethyl-53,54,55,56,57-pentaoxo-49-trimethylsilyloxy-72,73-dioxa-63-azatricyclohexatriaconta-29,31,33(51),34(52)-tetraen-48-yl]propyl]-46-methoxy-44-cyclohexyl]N-(2-morpholinoethyl)carbamate. (25E,27E,29E,30E,32R,33S,34R,35R,37S,39S,41S,42S,43R,44R,53R)-53-hydroxy-42-[(1R)-2-[(1S,3R,4R)-4-hydroxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-41,44-dimethoxy-32,33,34,35,45,46-hexamethyl-43-thiazolinone in DCM (40 mL) To a solution of 2,63-trimethylsilyloxy-62,63-dioxa-54-azatricyclohexatriaconta-25,27,29(45),30(46)-tetraene-47,48,49,50,51-pentone (2 g, 2.03 mmol) and pyridine (0.64 g, 8.11 mmol) was added triphosgene (0.6 g, 2.03 mmol) in 10 mL of DCM dropwise via syringe at 0° C. under argon. The reaction mixture was stirred at 0° C. for 1 h, and then triethylaminoethanol (TEA) (0.62 g, 6.08 mmol) and 2-morpholinoethanamine (0.53 g, 4.06 mmol) were added to the mixture. The resulting solution was stirred at 0° C. for an additional 1 h, then diluted with DCM, washed with aqueous NH4Cl, water, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel chromatography (8% MeOH in DCM) to give the title compound (2 g, 86% yield) as a pale yellow solid.

[0530] Step 3: [(39S,41R,43R)-4-[(2R)-2-[(26E,28E,30E,31E,35R,36S,37R,38R,40S,42S,44S,45S,46R,47R,57R)-46,57-dihydroxy-44,47-dimethoxy-35,36,37,38,48,49-hexamethyl-50,51,52,53,54-pentaoxo-70,71-dioxa-60-azatricyclohexatriaconta-26,28,30(48),31(49)-tetraen-45-yl]propyl]-43-methoxy-41-cyclohexyl]N-(2-morpholinoethyl)carbamate. [(42S,44R,46R)-4-[(2R)-2-[(29E,31E,33E,34E,38R,39S,40R,41R,43S,45S,47S,48S,49R,50R,60R)-60-hydroxy-47,50-dimethoxy-38,39,40,41,51,52-hexamethyl-53,54,55,56,57-pentaoxo- To a solution of 49-trimethylsilyloxy-72,73-dioxa-63-azatricyclohexatriaconta-29,31,33(51),34(52)-tetraen-48-yl]propyl]-46-methoxy-44-cyclohexyl]-N-(2-morpholinoethyl)carbamate (2 g, 1.75 mmol) was added 0.5 N H2SO4 (2.63 mmol, 5.3 mL) at 0 °C. The resulting solution was stirred at 0 °C for 8 h and then poured into a mixture of 100 mL of EtOAc and 100 mL of saturated aqueous NaHCO3. The organic layer was washed with water and brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel chromatography (5% MeOH in DCM) to give the title compound (1.5 g, 80% yield) as a pale yellow solid.

[0531] Step 4: [(42S,44R,46R)-4-[(2R)-2-[(27E,29E,31E,32E,38R,39S,40R,41R,43S,45S,48S,50R,51R,61R)-50,61-dihydroxy-51-methoxy-38,39,40,41,52,53-hexamethyl-47-[2-(oxetamethyl)-2-methyl-4-(2-hydroxyethyl)-2-methyl-4-(2-methyl-4-methyl- ...

[0033] N-(2-morpholinoethyl)carbamate. [(39S,41R,43R)-4-[(2R)-2-[(26E,28E,30E,31E,35R,36S,37R,38R,40S,42S,44S,45S,46R,47R,57R)-46,57-dihydroxy-44,47-dimethoxy-35,36,37,38,48,49-hexamethyl-50,51,52,53,54-pentamethyl-4-methyl-2-methyl-3-methyl-4-methyl-2 ... To a solution of oxo-70,71-dioxa-60-azatricyclohexatriaconta-26,28,30(48),31(49)-tetraen-45-yl]propyl]-43-methoxy-41-cyclohexyl]N-(2-morpholinoethyl)carbamate (0.2 g, 0.18 mmol) under nitrogen at −40° C., TFA (0.85 g, 7.47 mmol) was added. 2-(Oxetan-3-yloxy)ethanol (0.22 g, 1.87 mmol) was added, and the mixture was stirred at −30° C. for 2 hours. The mixture was poured into cold saturated aqueous NaHCO (30 mL), extracted with DCM (30 mL), washed with water (30 mL) and brine (30 mL), dried over anhydrous NaSO, filtered, and concentrated. The residue was purified by reverse-phase column eluting with 80% CH3CN in water to give I-19 (35 mg, 16% yield) as a white solid. ESI-MS (EI + , m / z): 1179.6 [M + Na] + . 1H NMR(400MHz,CDCl3)δ 6.41-5.90(m,4H),5.58-5.39(m,2H),5.30-5.15(m,2H),4.80-4.51(m,5H),4.32 -3.95(m,2H),3.92-3.66(m,7H),3.60-3.40(m,4H),3.39-3.20(m,11H),3.19-3.0 5(m,2H),2.79-2.62(m,2H),2.61-2.40(m,7H),2.37-2.20(m,2H),2.15-1.90(m,5 H),1.84-1.58(m,17H),1.54-1.16(m,7H),1.15-0.83(m,17H),0.82-0.75(m,1H).

[0532] Step 5: [(42S,44R,46R)-4-[(2R)-2-[(27E,29E,31E,32E,38R,39S,40R,41R,43S,45S,47S,48S,50R,51R,61R)-50,61-dihydroxy-51-methoxy-38,39,40,41,52,53-hexamethyl-47-[2-(oxomethyl)amino]methyl] 54,55,56,57,58-pentaoxo-75,76-dioxa-64-azatricyclohexatriaconta-27,29,31(52),32(53)-tetraen-48-yl]propyl]-46-methoxy-44-cyclohexyl]N-(2-morpholinoethyl)carbamate (I-2 1), and [(42S,44R,46R)-4-[(2R)-2-[(27E,29E,31E,32E,38R,39S,40R,41R,43S,45S,47R,48S,50R,51R,61R)-50,61-dihydroxy-51-methoxy-38,39,40,41,52,53-hexamethyl-47-[2-(oxetane -3-yloxy)ethoxy]-54,55,56,57,58-pentaoxo-75,76-dioxa-64-azatricyclohexatriaconta-27,29,31(52),32(53)-tetraen-48-yl]propyl]-46-methoxy-44-cyclohexyl]N-(2-morpholinoethyl)carbamate (I-20)

[0533] 115 mg of [(42S,44R,46R)-4-[(2R)-2-[(27E,29E,31E,32E,38R,39S,40R,41R,43S,45S,48S,50R,51R,61R)]-50,61-dihydroxy-51-methoxy-38,39,40,41,52,53-hexamethyl-47-[2-(oxetan-3-yloxy)ethoxy]-54,55,56,57,58-pentaoxo-75,76-dioxa-64-azatricyclohexyl The resulting tetraconta-27,29,31(52),32(53)-tetraen-48-yl]propyl]-46-methoxy-44-cyclohexyl]N-(2-morpholinoethyl)carbamate was separated by chiral preparative HPLC and then purified by silica gel chromatography (12% MeOH in petroleum ether:DCM:EA=3:3:1) to give I-21 (23 mg, 20% yield) as a white solid and I-20 (10 mg, 8.7% yield) as a white solid.

[0534] Chiral separation methods: Column: CHIRALPAK IC Column size: 2.5cm inner diameter x 25cm length, 10μm Sample solution: 3mg / mL in mobile phase Injection: 10mL Mobile phase: EtOH=100% Flow rate: 10mL / min Wavelength: UV254nm Temperature: 35℃ I-21: ESI-MS (EI + , m / z): 1156.9 [M + H] + ,1178.8[M+Na] + . 1 H NMR(400MHz,CDCl3)δ 6.51-5.78(m,4H),5.74-5.01(m,5H),4.68(ddd,J=37.1,11.1,6.1Hz,6H),4.32-4.13( m, 1H), 3.96-3.06 (m, 23H), 2.79-2.24 (m, 10H), 2.17-1.18 (m, 29H), 1.17-0.76 (m, 19H). I-20: ESI-MS (EI +, m / z): 1156.9 [M + H] + ,1178.8[M+Na] + . 1 H NMR(400MHz,CDCl3)δ 6.58-5.81(m,4H),5.67-5.02(m,5H),4.44(dd,J=176.8,44.7Hz,6H),4.03 -3.05(m,26H),2.83-2.29(m,10H),2.17-1.19(m,27H),1.16-0.69(m,19H).

[0535] Example 13: (22E,24E,26E,27E,35R,36S,37R,38R,40S,42S,45S,46R,47R,56R)-44-[2-(2-aminoethoxy)ethoxy]-56-hydroxy-45-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-46,47-dimethoxy-35,36,37,38,48,49-hexamethyl-66,67-dioxa-58-azatricyclohexatriaconta-22,24,26(48),27(49)-tetraene-50,51,52,53,54-pentone (I-23)

[0536] [ka]

[0537] Step 1: (36E,38E,40E,41E,47R,48S,49R,50R,52S,54S,56S,57S,58R,59R,68R)-57-[(1R)-2-[(1S,3R,4R)-4-[2-[tert-butyl(diphenyl)silyl]oxyethoxy]-3-methoxy-cyclohexyl]-1-methyl-ethyl]-68-hydroxy-56,58,59-trimethoxy-47,48,49,50,60,61-hexamethyl-77,78-dioxa-70-azatricyclohexatriaconta-36,38,40(60),41(61)-tetraene-62,63,64,65,66-pentone. (35E,37E,39E,40E,46R,47S,48R,49R,51S,53S,55S,56S,57R,58R,67R)-56-[(1R)-2-[(1S,3R,4R)-4-[2-[tert-butyl(diphenyl)silyl]oxyethoxy]-3-methoxy-cyclohexyl]-1-methyl-ethyl]-57,67-dihydroxy-55,58-dimethoxy-46,47,48,49,59,60-hexamethyl-4-(2-tert-butyl(diphenyl)silyl) ... To a solution of methyl-77,78-dioxa-69-azatricyclohexatriaconta-35,37,39(59),40(60)-tetraene-61,62,63,64,65-pentone (Intermediate IX prepared according to Example 22, 2 g, 1.67 mmol) and 1,8-bis(dimethylamino)naphthalene (3.94 g, 18.39 mmol), methyl trifluoromethanesulfonate (2.19 g, 13.37 mmol) was added dropwise at room temperature under N. After the addition, the mixture was heated to 50 °C for 5 h, then washed, filtered, diluted with EA (60 mL), and washed with saturated NH4Cl(aq) (60 mL × 3), water (60 mL), and brine (60 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel chromatography (petroleum ether:EA=3:1) to give the title compound (700 mg, 35% yield) as a yellow solid. + , m / z): 1232.7 [M + Na] + .

[0538] Step 2: (23E,25E,27E,28E,32R,33S,34R,35R,37S,39S,41S,42S,43R,44R,53R)-53-hydroxy-42-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-41,43,44-trimethoxy-32,33,34,35,45,46-hexamethyl-62,63-dioxa-54-azatricyclohexatriaconta-23,25,27(45),28(46)-tetraene-47,48,49,50,51-pentone. (36E,38E,40E,41E,47R,48S,49R,50R,52S,54S,56S,57S,58R,59R,68R)-57-[(1R)-2-[(1S,3R,4R)-4-[2-[tert-butyl(diphenyl)silyl]oxyethoxy]-3-methoxy-cyclohexyl]-1-methyl-ethyl]-68-hydroxy-56, To a solution of 58,59-trimethoxy-47,48,49,50,60,61-hexamethyl-77,78-dioxa-70-azatricyclohexatriaconta-36,38,40(60),41(61)-tetraene-62,63,64,65,66-pentone (600 mg, 0.496 mmol) was added pyridine HF (392 mg, 4.96 mmol) at 0 °C. The mixture was stirred at 30 °C for 3 h, then quenched by the addition of saturated aqueous NaHCO (20 mL) and extracted with EA (30 mL) at 0 °C. The organic layer was washed with water (20 mL) and brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel chromatography (petroleum ether:acetone=3:1) to give the title compound (430 mg, 89% yield) as a pale yellow solid. + , m / z): 994.7 [M + Na] + .

[0539] Step 3: (22E,24E,26E,27E,35R,36S,37R,38R,40S,42S,45S,46R,47R,56R)-44-[2-(2-azidoethoxy)ethoxy]-56-hydroxy-45-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-46,47-dimethoxy-35,36,37,38,48,49-hexamethyl-68,69-dioxa-60-azatricyclohexatriaconta-22,24,26(48),27(49)-tetraene-50,51,52,53,54-pentone. (23E,25E,27E,28E,32R,33S,34R,35R,37S,39S,41S,42S,43R,44R,53R)-53-hydroxy-42-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-41,43,44-trimethoxy-32,33,34,35, To a solution of 45,46-hexamethyl-62,63-dioxa-54-azatricyclohexatriaconta-23,25,27(45),28(46)-tetraene-47,48,49,50,51-pentone (450 mg, 462.85 μmol) and 2-(2-methoxyethoxy)ethanol (1.21 g, 9.26 mmol) was added HND-8 (100 mg) under N at 50 °C. The reaction mixture was stirred at 50 °C for 22 h, then cooled and filtered. The filtrate was poured into saturated aqueous NaHCO (10 mL) at 0 °C and extracted with EA (30 mL). The organic layer was washed with water (10 mL) and brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by reverse phase chromatography (70% CH3CN in water) to give the title compound (250 mg, 50% yield) as a pale yellow solid. ESI-MS (EI + , m / z): 1093.4 [M + Na] + .

[0540] Step 4: (22E,24E,26E,27E,35R,36S,37R,38R,40S,42S,45S,46R,47R,56R)-44-[2-(2-aminoethoxy)ethoxy]-56-hydroxy-45-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-46,47-dimethoxy-35,36,37,38,48,49-hexamethyl-66,67-dioxa-58-azatricyclohexatriaconta-22,24,26(48),27(49)-tetraene-50,51,52,53,54-pentone (I-23). (22E,24E,26E,27E,35R,36S,37R,38R,40S,42S,45S,46R,47R,56R) 44-[2-(2-azidoethoxy)ethoxy]-56-hydroxy-45-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-46 in THF (10 mL) To a solution of 1-47-dimethoxy-35,36,37,38,48,49-hexamethyl-68,69-dioxa-60-azatricyclohexatriaconta-22,24,26(48),27(49)-tetraene-50,51,52,53,54-pentone (0.6 g, 0.56 mmol), triphenylphosphine (0.44 g, 1.68 mmol) was slowly added. The resulting solution was stirred at 60 °C for 2 h, 0.05 mL of water was added, and the reaction was stirred at room temperature for 6 h and then concentrated. The residue was purified by reverse-phase chromatography (CHCN / HO with 0.025% TFA) to give I-23 (40 mg, 7% yield) as a white solid. ESI-MS (EI + , m / z): 1045.7 [M + H] + . 1H NMR(400MHz,CDCl3)δ 4.61-5.96(m,4H),5.69-5.07(m,4H),4.51-4.01(m,3H),3.82-3.52(m,7H),3.47-3.37(m,5H),3.31-3.04(m,13H),2.88-2.5 2(m,2H),2.38-1.97(m,7H),1.85-1.52(m,17H),1.38-1.13(m,7H),1.12-0.98(m,5H),0.98-0.77(m,17H),0.75-0.68(m,1H).

[0541] Example 14: (1R,2R,4S)-4-((2R)-2-((3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-21-((1,4-dioxan-2-yl)methoxy)-27-hydroxy-9,10-dimethoxy-6,8,12,14,20,26-hexamethyl-1,5 ,11,28,29-Pentaoxo-1,4,5,6,9,10,11,12,13,14,21,22,23,24,25,26,27,28,29,31,32,33,34,34a-tetracosahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohentriacontin-3-yl)propyl)-2-meth cyclohexyl dimethyl phosphinate (I-24), and (27E,29E,31E,32E,35R,36S,37R,38R,40S,42S,44S,45S,47R,48R,57R)-45-[(1R)-2-[(1S,3R,4R)-4-dimethylphosphoryloxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]- Synthesis of 44-(1,4-dioxan-2-ylmethoxy)-57-hydroxy-47,48-dimethoxy-35,36,37,38,49,50-hexamethyl-68,69-dioxa-58-azatricyclohexatriaconta-27,29,31(49),32(50)-tetraene-51,52,53,54,55-pentone (I-25).

[0542] [ka]

[0543] Step 1: (3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-3-((R)-1-((1S,3R,4R)-4-((tert-butyldimethylsilyl)oxy)-3-methoxycyclohexyl)propan-2-yl)-9,27-dihydroxy-10,21-dimethoxy To a solution of rapamycin (5 g, 5.47 mmol) in DMF (60 mL) at room temperature, imidazole (1.49 g, 21.88 mmol) and tert-butyl-chloro-dimethyl-silane (2.47 g, 16.41 mmol) were added. The reaction was stirred at 50 °C for 6 h and then poured into a mixture of ice-cold saturated aqueous NH4Cl (40 mL) and Et2O:petroleum ether (60 mL, 2:1). The organic layer was washed with saturated aqueous NH4Cl (20 mL), water and brine (20 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel chromatography (10% to 50% EtOAc in petroleum ether) to give the title compound (4 g, 71% yield) as a white solid. ESI-MS (EI + , m / z): 1050.5 [M + Na] + .

[0544] Step 2: (3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-3-((R)-1-((1S,3R,4R)-4-((tert-butyldimethylsilyl)oxy)-3-methoxycyclohexyl)propan-2-yl)-27-hydroxy-9,10,21-trimethoxysilane Ci-6,8,12,14,20,26-hexamethyl-9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-hexadecahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohentriacontin-1,5,11,28,29(4H,6H,31H)-pentaone. (3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-3-((R)-1-((1S,3R,4R)-4-((tert-butyldimethylsilyl)oxy)-3-methoxycyclohexyl)propan-2-yl)-9,27-dihydroxy-10,21-dimethoxy-6,8,12,14,20,26-hexamethyl-9,10,12,13,14,21,22,23,2 To a suspension of 4,25,26,27,32,33,34,34a-hexadecahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohentriacontin-1,5,11,28,29(4H,6H,31H)-pentaone (1 g, 0.97 mmol) and 1,8-bis(dimethylamino)naphthalene (2.5 g, 11.67 mmol), methyl trifluoromethanesulfonate (1.6 mL, 14.59 mmol) was added dropwise at room temperature under N. After the addition, the mixture was heated to 50 °C for 6 h, then cooled and filtered. The filtrate was purified by silica gel chromatography to give the title compound (0.45 g, 0.43 mmol) as a white solid. ESI-MS (EI+, m / z): 1064.6 [M+Na] + .

[0545] Step 3: (3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-27-hydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propan-2-yl)-9,10,21-trimethoxy-6,8,12, 14,20,26-Hexamethyl-9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-hexadecahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohentriacontin-1,5,11,28,29(4H,6H,31H)-pentaone. (3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-3-((R)-1-((1S,3R,4R)-4-((tert-butyldimethylsilyl)oxy)-3-methoxycyclohexyl)propan-2-yl)-27-hydroxy-9,10,21-trimethoxy-6,8,12,14,20,26-hexamethyl-9, To a solution of 10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-hexadecahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohentriacontin-1,5,11,28,29(4H,6H,31H)-pentaone (400 mg, 0.38 mmol) was added pyridine hydrofluoride (3.34 mL, 38.37 mmol) at 0 °C. The reaction was warmed to 45 °C and stirred for 5 h, then diluted with DCM and aqueous NaHCO, washed with water and brine, dried over NaSO, filtered, and concentrated. The residue was purified by silica gel chromatography to give the title compound (0.16 g, 45% yield) as a white solid. ESI-MS(EI + , m / z): 949.9 [M + Na] + .

[0546] Step 4: (1R,2R,4S)-4-((R)-2-((3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-27-hydroxy-9,10,21-trimethoxy-6,8,12,14,20,26-hexamethyl-1,5,11,28,29-pentaoxo-1,4 ,5,6,9,10,11,12,13,14,21,22,23,24,25,26,27,28,29,31,32,33,34,34a-Tetracosahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohentriacontin-3-yl)propyl)-2-methoxycyclohexyldimethylphosphinate. (3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-27-hydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propan-2-yl)-9,10,21-trimethoxy-6,8,12,14,20,26-hexamethyl-9,10,12,13,14,21,22,23,24,25,26,27,3 To a solution of 2,33,34,34a-hexadecahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohentriacontin-1,5,11,28,29(4H,6H,31H)-pentaone (0.26 g, 0.28 mmol) in DCM (1 mL) was added 2,6-di-tert-butyl-4-methylpyridine (0.173 g, 0.84 mmol) and dimethylphosphinic chloride (0.315 g, 2.8 mmol) at 0° C. The resulting solution was stirred at 0° C. for 5 h, then diluted with EtOAc, washed with NaHCO solution, ice-cold 0.5 N HCl, water, brine, dried over NaSO, filtered, and concentrated. The residue was purified by silica gel chromatography (DCM:MeOH=40:1) to give the title compound (0.1 g, 36% yield) as a white solid. + , m / z): 1025.8 [M + Na] + .

[0547] Step 5: (1R,2R,4S)-4-((2R)-2-((3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-21-((1,4-dioxan-2-yl)methoxy)-27-hydroxy-9,10-dimethoxy-6,8,12,14,20,26-hexamethyl-1,5,11,28,29-pentaenoic acid Xo-1,4,5,6,9,10,11,12,13,14,21,22,23,24,25,26,27,28,29,31,32,33,34,34a-tetracosahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohentriacontin-3-yl)propyl)-2-methoxycyclohexyldimethylphosphinate (I-24). (1R,2R,4S)-4-((R)-2-((3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-27-hydroxy-9,10,21-trimethoxy-6,8,12,14,20,26-hexamethyl-1,5,11,28,29-pentaoxo-1,4,5,6,9,10,11,12,13,14,21, To a solution of 22,23,24,25,26,27,28,29,31,32,33,34,34a-tetracosahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohentriacontin-3-yl)propyl)-2-methoxycyclohexyldimethylphosphinate (129 mg, 0.129 mmol) was added TFA (0.49 mL, 6.42 mmol) at -50 °C. The mixture was stirred at the same temperature for 10 min, and then 2-(oxetan-3-yloxy)ethanol (0.758 g, 6.42 mmol) dissolved in DCM (0.5 mL) was added, and the mixture was stirred at 0 °C for 6 h. The reaction was then diluted with DCM and aqueous NaHCO, washed with water and brine, dried over NaSO, filtered, and concentrated. The residue was purified by reverse phase chromatography (70% CHCN in water) to give I-24 (30 mg, 21% yield) as a white solid. + , m / z): 1112.8 [M + Na] + . 1H NMR(400MHz,CDCl3)δ 6.49(br,4H),5.63-5.06(m,4H),4.78-4.05(m,3H),3.66(ddd,J=23.6,18.4,8.5Hz,9H),3.49 -2.97(m,17H),2.82-2.48(m,2H),2.37-1.86(m,7H),1.56-1.23(m,22H),1.18-0.68(m,24H).

[0548] Step 6: (27E,29E,31E,32E,35R,36S,37R,38R,40S,42S,44S,45S,47R,48R,57R)-45-[(1R)-2-[(1S,3R,4R)-4-dimethylphosphoryloxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-44-(1,4-dioxan-2-ylmethoxy)-57-hydroxy-47,48-dimethoxy-35,36,37,38,49,50-hexamethyl-68,69-dioxa-58-azatricyclohexatriaconta-27,29,31(49),32(50)-tetraene-51,52,53,54,55-pentone (I-25). 100 mg of I-24 was separated by chiral preparative HPLC and then purified by silica gel chromatography (8% MeOH in petroleum ether:DCM:EA:3:3:1) to give I-25 (14 mg, 14% yield) as a white solid.

[0549] Chiral separation methods: Column: CHIRALPAK IC Column size: 2.5cm inner diameter x 25cm length, 10μm Sample solution: 9 mg / mL in mobile phase Injection: 15mL Mobile phase: Hexane / EtOH = 50 / 50 (V / V) Flow rate: 30mL / min Wavelength: UV254nm Temperature: 35℃ ESI-MS(EI + , m / z): 1112.6 [M + Na] + . 1H NMR(400MHz,CDCl3)δ 6.49-5.80(m,4H),5.67-5.15(m,4H),4.20-4.02(m,2H),3.98-3.55(m,12H),3.52-3.00(m,17H),2.60(ddd,J=39 .8,34.1,28.4Hz,6H),2.37-1.83(m,7H),1.66(dt,J=39.0,20.7Hz,12H),1.42-1.19(m,8H),1.18-0.66(m,20H).

[0550] Example 15: (25E,27E,29E,30E,33R,34S,35R,36R,38S,40S,43S,45R,46R,55R)-42-(1,4-dioxan-2-ylmethoxy)-55-hydroxy-43-[(1R)-2-[(1S,3R,4R)-4-hydroxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-45,46-dimethoxy-33,34,35,36,47,48-hexamethyl-66,67-dioxa-56-azatricyclohexatriaconta-25,27,29(47),30(48)-tetraene-49,50,51,52,53-pentone (I-26), and Synthesis of (25E,27E,29E,30E,33R,34S,35R,36R,38S,40S,42S,43S,45R,46R,55R)-42-(1,4-dioxan-2-ylmethoxy)-55-hydroxy-43-[(1R)-2-[(1S,3R,4R)-4-hydroxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-45,46-dimethoxy-33,34,35,36,47,48-hexamethyl-66,67-dioxa-56-azatricyclohexatriaconta-25,27,29(47),30(48)-tetraene-49,50,51,52,53-pentone (I-27)

[0551] [ka]

[0552] Step 1: (25E,27E,29E,30E,33R,34S,35R,36R,38S,40S,43S,45R,46R,55R)-42-(1,4-dioxan-2-ylmethoxy)-55-hydroxy-43-[(1R)-2-[(1S,3R,4R)-4-hydroxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-45,46-dimethoxy-33,34,35,36,47,48-hexamethyl-66,67-dioxa-56-azatricyclohexatriaconta-25,27,29(47),30(48)-tetraene-49,50,51,52,53-pentone (I-26). (23E,25E,27E,28E,30R,31S,32R,33R,35S,37S,39S,40S,41R,42R,51R)-51-hydroxy-40-[(1R)-2-[(1S,3R,4R)-4-hydroxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-39,41,42-trimethoxy-30,31,32,33,43,44-hexamethyl-6 in THF (15 mL) To a solution of 0,61-dioxa-52-azatricyclohexatriaconta-23,25,27(43),28(44)-tetraene-45,46,47,48,49-pentone (Intermediate X prepared according to Example 16, 0.312 g, 0.336 mmol), 2-(oxetan-3-yloxy)ethanol (0.397 g, 3.36 mmol) and HND-8 (624 mg) were added under nitrogen at 0 °C. The mixture was stirred at 50 °C for 5 h and then purified by reverse-phase chromatography eluting with 80% CHCN in water followed by preparative TLC (petroleum ether: EtOAc = 1:2) to give I-26 (30 mg, 9% yield) as a white solid. ESI-MS (EI + , m / z): 1035.8 [M + Na] + . 1 H NMR (500 MHz, CDCl): 1H NMR(500MHz,CDCl3)δ 6.57-5.90(m,3H),5.71-5.00(m,3H),4.72-4.10(m,1H),3.91-3.52(m,7H),3.38(dd,J=22.8,12.9Hz,5H),3.30-3.15(m,3H),3.16-3.02(m ,3H),3.00-2.46(m,4H),2.15(dd,J=97.2,37.0Hz,5H),1.85-1.53(m,23H),1.52-1.21(m,9H),1.19-0.82(m,14H),0.69(d,J=11.9Hz,1H).

[0553] Step 2: (25E,27E,29E,30E,33R,34S,35R,36R,38S,40S,42S,43S,45R,46R,55R)-42-(1,4-dioxan-2-ylmethoxy)-55-hydroxy-43-[(1R)-2-[(1S,3R,4R)-4-hydroxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-45,46-dimethoxy-33,34,35,36,47,48-hexamethyl-66,67-dioxa-56-azatricyclohexatriaconta-25,27,29(47),30(48)-tetraene-49,50,51,52,53-pentone (I-27). 85 mg of the epimeric mixture was purified by preparative chiral HPLC, followed by silica gel chromatography (hexane:DCM:EtOAc:MeOH=3:3:1:0.3) to afford I-27 (25 mg, 29% yield) as a white solid.

[0554] Chiral separation methods: Column: CHIRALPAK IC Column size: 5.0 cm inner diameter x 25 cm length, 10 μm Sample solution: 0.3mg / mL in mobile phase Injection: 3mL Mobile phase: hexane / EtOH = 70 / 30 (V / V) Flow rate: 25mL / min Wavelength: UV254nm Temperature: 35℃ ESI-MS(EI +, m / z): 1036.4 [M + Na] + . 1 H NMR(400MHz,CDCl3)δ 6.44-5.80(m,4H),5.65-5.01(m,4H),4.64(d,J=15.9Hz,1H),3.99-3.52(m,11H),3.47-3.02(m,16H),3.02-2 .46(m,5H),2.43-1.85(m,8H),1.83-1.64(m,9H),1.46-1.19(m,10H),1.16-0.83(m,18H),0.79-0.59(m,1H).

[0555] Example 16: Synthesis of (23E,25E,27E,28E,36R,37S,38R,39R,41S,43S,46S,47R,48R,57R)-57-hydroxy-46-[(1R)-2-[(1S,3R,4R)-4-hydroxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-47,48-dimethoxy-45-[2-[2-(2-methoxyethoxy)ethoxy]ethoxy]-36,37,38,39,49,50-hexamethyl-66,67-dioxa-58-azatricyclohexatriaconta-23,25,27(49),28(50)-tetraene-51,52,53,54,55-pentone (I-28):

[0556] [ka]

[0557] Step 1: (27E,29E,31E,32E,34R,35S,36R,37R,39S,41S,43S,44S,45R,46R,55R)-44-[(1R)-2-[(1S,3R,4R)-4-[tert-butyl(dimethyl)silyl]oxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-45,55-dihydroxy-43,46-dimethoxy-34,35,36,37,47,48-hexamethyl-65,66-dioxa-57-azatricyclohexatriaconta-27,29,31(47),32(48)-tetraene-49,50,51,52,53-pentone. (22E,24E,26E,27E,29R,30S,31R,32R,34S,36S,38S,39S,40R,41R,50R)-40,50-dihydroxy-39-[(1R)-2-[(1S,3R,4R)-4-hydroxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-38,41-dimethoxy-29,30,31,32,42,4 To a solution of 3-hexamethyl-60,61-dioxa-51-azatricyclohexatriaconta-22,24,26(42),27(43)-tetraene-44,45,46,47,48-pentone (2 g, 2.19 mmol), imidazole (0.596 g, 8.75 mmol) and tert-butylchlorodimethylsilane (0.989 g, 6.56 mmol) were added at room temperature. The mixture was stirred at 20 °C for 5 h and then poured into ice-cold saturated aqueous NH4Cl (40 mL) and Et2O:petroleum ether (60 mL, 2:1). The organic layer was washed with saturated aqueous NH4Cl (20 mL), water, and brine (20 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel chromatography (10% to 50% EtOAc in petroleum ether) to give the title compound (1.5 g, 67% yield) as a white solid. + , m / z): 1049.8 [M + Na] + .

[0558] Step 2: (28E,30E,32E,33E,35R,36S,37R,38R,40S,42S,44S,45S,46R,47R,56R)-45-[(1R)-2-[(1S,3R,4R)-4-[tert-butyl(dimethyl)silyl]oxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-56-hydroxy-44,46,47-trimethoxy-35,36,37,38,48,49-hexamethyl-65,66-dioxa-58-azatricyclohexatriaconta-28,30,32(48),33(49)-tetraene-50,51,52,53,54-pentone. (27E,29E,31E,32E,34R,35S,36R,37R,39S,41S,43S,44S,45R,46R,55R)-44-[(1R)-2-[(1S,3R,4R)-4-[tert-butyl(dimethyl)silyl]oxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-45,55-dihydroxy-43,46-dimethoxy-34,35,36,37,47, To a suspension of 48-hexamethyl-65,66-dioxa-57-azatricyclohexatriaconta-27,29,31(47),32(48)-tetraene-49,50,51,52,53-pentone (0.6 g, 0.58 mmol) and 1,8-bis(dimethylamino)naphthalene (1.5 g, 7 mmol) was added methyl trifluoromethanesulfonate (0.957 g, 5.83 mmol) dropwise at room temperature under N2. After the addition, the mixture was heated to 50 °C for 6 h, then cooled and filtered. The filtrate was purified by silica gel chromatography (EtOAc:petroleum ether = 4:1) to give the title compound (0.24 g, 39% yield) as a white solid. ESI-MS (EI + , m / z): 1063.8 [M + Na] + .

[0559] Step 3: (23E,25E,27E,28E,30R,31S,32R,33R,35S,37S,39S,40S,41R,42R,51R)-51-hydroxy-40-[(1R)-2-[(1S,3R,4R)-4-hydroxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-39,41,42-trimethoxy-30,31,32,33,43,44-hexamethyl-60,61-dioxa-52-azatricyclohexatriaconta-23,25,27(43),28(44)-tetraene-45,46,47,48,49-pentone. (28E,30E,32E,33E,35R,36S,37R,38R,40S,42S,44S,45S,46R,47R,56R)-45-[(1R)-2-[(1S,3R,4R)-4-[tert-butyl(dimethyl)silyl]oxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-56-hydroxy-44,46,47-trimethyl-4-(2-(tert-butyl(dimethyl)silyl)oxy) ... To a solution of methoxy-35,36,37,38,48,49-hexamethyl-65,66-dioxa-58-azatricyclohexatriaconta-28,30,32(48),33(49)-tetraene-50,51,52,53,54-pentone (0.24 g, 0.23 mmol) was added hydrogen fluoride pyridine (2.28 g, 23.02 mmol, 2 mL) at 0 °C. The reaction was stirred at 45 °C for 5 h, then diluted with DCM and saturated aqueous NaHCO, washed with water and brine, dried over NaSO, filtered, and concentrated. The residue was purified by reverse-phase chromatography (78% CHCN in water) to give the title compound (0.105 g, 49% yield) as a white solid. ESI-MS (EI + , m / z): 949.7 [M + Na] + .

[0560] Step 4: (23E,25E,27E,28E,36R,37S,38R,39R,41S,43S,46S,47R,48R,57R)-57-hydroxy-46-[(1R)-2-[(1S,3R,4R)-4-hydroxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-47,48-dimethoxy-45-[2-[2-(2-methoxyethoxy)ethoxy]ethoxy]-36,37,38,39,49,50-hexamethyl-66,67-dioxa-58-azatricyclohexatriaconta-23,25,27(49),28(50)-tetraene-51,52,53,54,55-pentone (I-28). (23E,25E,27E,28E,30R,31S,32R,33R,35S,37S,39S,40S,41R,42R,51R)-51-hydroxy-40-[(1R)-2-[(1S,3R,4R)-4-hydroxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-39,41,42-trimethoxy-30,31,32,33,43,44-hexamethyl-6 in THF (15 mL) To a solution of 0,61-dioxa-52-azatricyclohexatriaconta-23,25,27(43),28(44)-tetraene-45,46,47,48,49-pentone (0.15 g, 0.16 mmol) under nitrogen at 0°C, 2-(2-(2-methoxyethoxy)ethoxy)ethanol (0.265 g, 1.62 mmol) and HND-8 (0.3 g) were added, and the mixture was stirred at 50°C for 8 hours.The reaction mixture was purified by reverse phase chromatography eluting with 80% CH3CN in water followed by preparative TLC (petroleum ether:ethyl acetate=1:2) to give (23E,25E,27E,28E,36R,37S,38R,39R,41S,43S,46S,47R,48R,57R)-57-hydroxy-46-[(1R)-2-[(1S,3R,4R)-4-hydroxy-3-methoxy-cyclohexyl]

[0049] -1-methyl-ethyl]-47,48-dimethoxy-45-[2-[2-(2-methoxyethoxy)ethoxy]ethoxy]-36,37,38,39,49,50-hexamethyl-66,67-dioxa-58-azatricyclohexatriaconta-23,25,27(49),28(50)-tetraene-51,52,53,54,55-pentone (36.5 mg, 21% yield) was obtained as a white solid. ESI-MS (EI). + , m / z): 1035.8 [M + Na] + . 1 H NMR (500 MHz, CDCl): 1 H NMR(400MHz,CDCl3)δ 6.59-5.88(m,3H),5.85-4.93(m,4H),4.72-4.18(m,1H),4.15-3.76(m,2H),3.74-3.52(m,8H),3.50-3.30(m,8H),3.29-3.03(m,5H),3. 03-2.47(m,5H),2.45-1.89(m,6H),1.90-1.52(m,21H),1.32(ddd,J=28.1,22.9,5.8Hz,9H),1.19-0.78(m,14H),0.69(d,J=12.0Hz,1H).

[0561] Example 17: (24E,26E,28E,29E,35R,36S,37R,38R,40S,42S,45S,46R,47R,56R)-45-[(1R)-2-[(1S,3R,4R)-3,4-dimethoxycyclohexyl]-1-methyl-ethyl]-56-hydroxy-46,47-dimethoxy-44-[2-(2-methoxyethoxy)ethoxy]-35,36,37,38,48,49-hexamethyl-64,65-dioxa-57-a Zatricyclohexatriaconta-24,26,28(48),29(49)-tetraene-50,51,52,53,54-pentone (I-29), (24E,26E,28E,29E,35R,36S,37R,38R,40S,42S,44S,45S,46R,47R,56R)-45-[(1R)-2-[(1S,3R,4R)-3,4-dimethoxycyclohexyl]-1-methyl-ethyl]-56-hydroxy-46,47-dimethoxy -44-[2-(2-methoxyethoxy)ethoxy]-35,36,37,38,48,49-hexamethyl-64,65-dioxa-57-azatricyclohexatriaconta-24,26,28(48),29(49)-tetraene-50,51,52,53,54-pentone (I-31), and (24E,26E,28E,29E,35R,36S,37R,38R,40S,42S,44R,45S,46R,47R,56R)-45-[( Synthesis of 1R)-2-[(1S,3R,4R)-3,4-dimethoxycyclohexyl]-1-methyl-ethyl]-56-hydroxy-46,47-dimethoxy-44-[2-(2-methoxyethoxy)ethoxy]-35,36,37,38,48,49-hexamethyl-64,65-dioxa-57-azatricyclohexatriaconta-24,26,28(48),29(49)-tetraene-50,51,52,53,54-pentone (I-30)

[0562] [ka]

[0563] Step 1: (24E,26E,28E,29E,31R,32S,33R,34R,36S,38S,40S,41S,42R,43R,52R)-41-[(1R)-2-[(1S,3R,4R)-3,4-dimethoxycyclohexyl]-1-methyl-ethyl]-52-hydroxy-40,42,43-trimethoxy-31,32,33,34,44,45-hexamethyl-60,61-dioxa-53-azatricyclohexatriaconta-24,26,28(44),29(45)-tetraene-46,47,48,49,50-pentone. (22E,24E,26E,27E,29R,30S,31R,32R,34S,36S,38S,39S,40R,41R,50R)-40,50-dihydroxy-39-[(1R)-2-[(1S,3R,4R)-4-hydroxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-38,41-dimethoxy-29,30,31,32,42,43- in toluene (5 mL) To a solution of hexamethyl-60,61-dioxa-51-azatricyclohexatriaconta-22,24,26(42),27(43)-tetraene-44,45,46,47,48-pentone (0.2 g, 0.219 mmol) was added proton sponge (0.938 g, 4.38 mmol), followed by methyl trifluoromethanesulfonate (0.539 g, 3.28 mmol). The mixture was stirred at 50 °C for 6 h, cooled, and purified by silica gel chromatography followed by reverse-phase chromatography (85% CH3CN in water) to give the title compound (50 mg, 24% yield) as a white solid. ESI-MS (EI + , m / z): 964.2 [M + Na] + . 1H NMR(400MHz,CDCl3)δ 6.50-5.80(m,4H),5.62(ddd,J=22.9,14.5,7.9Hz,1H),5.32(dt,J=11.6,7.7Hz,2H),5.18-5.03(m,1H),4.68(s,1H),3.95-3.54(m,5H),3.50-3.33(m,7H),3.32-3.21(m,3H),3.18-2.92(m,8H),2.83-2.48(m,3H),2.25(dd,J=30.1,10.7Hz,2H),2.02(ddd,J=34.0,26.3,9.6Hz,4H),1.88-1.56(m,14H),1.51-1.16(m,9H),1.15-0.82(m,18H),0.79-0.68(m,1H)。

[0564] Step 2: (24E,26E,28E,29E,35R,36S,37R,38R,40S,42S,45S,46R,47R,56R)-45-[(1R)-2-[(1S,3R,4R)-3,4-dimethoxycyclohexyl]-1-methyl-ethyl]-56-hydroxy-46,47-dimethoxy-44-[2-(2-methoxyethoxy)ethoxy]-35,36,37,38,48,49-hexamethyl-64,65-dioxa-57-azatricyclohexatriaconta-24,26,28(48),29(49)-tetraene-50,51,52,53,54-pentone (I-29). (24E,26E,28E,29E,31R,32S,33R,34R,36S,38S,40S,41S,42R,43R,52R)-41-[(1R)-2-[(1S,3R,4R)-3,4-dimethoxycyclohexyl]-1-methyl-ethyl]-52-hydroxy-40,42,43-trimethoxy-31,32,33,34,44,45-hexyl] in sulfolane (5 mL) To a solution of 1-hexamethyl-60,61-dioxa-53-azatricyclohexatriaconta-24,26,28(44),29(45)-tetraene-46,47,48,49,50-pentone (0.17 g, 0.18 mmol) and 2-(2-methoxyethoxy)ethoxyethanol (0.43 g, 3.61 mmol) was added HND-8 (35 mg) under N at 50 °C. The resulting solution was stirred at 50 °C for 3 h, filtered, and the filtrate was applied to a C18 column and eluted with 85% CH3CN in water to give I-29 as a white solid (65 mg, 35% yield). ESI-MS (EI + , m / z): 1052.5 [M + Na] + . 1 H NMR(500MHz,CDCl3)δ 6.46-5.81(m,4H),5.74-5.03(m,4H),4.68-4.15(m,2H),3.99-3.52(m,11H),3.50-3.22(m,16H),3.21-2.98(m,6H),2.94 -2.44(m,3H),2.37-1.89(m,7H),1.86-1.69(m,7H),1.52-1.24(m,9H),1.22-0.84(m,21H),0.74(dd,J=22.3,10.9Hz,1H).

[0565] Step 3: (24E,26E,28E,29E,35R,36S,37R,38R,40S,42S,44S,45S,46R,47R,56R)-45-[(1R)-2-[(1S,3R,4R)-3,4-dimethoxycyclohexyl]-1-methyl-ethyl]-56-hydroxy-46,47-dimethoxy-44-[2-(2-methoxyethoxy)ethoxy]-35,36,37,38,48,49-hexamethyl-64,65-dioxa-57-azatricyclohexatriaconta-24,26,28(48),29(49)-tetraene-50,51,52,53,54-pentone (I-3 1) and (24E,26E,28E,29E,35R,36S,37R,38R,40S,42S,44R,45S,46R,47R,56R)-45-[(1R)-2-[(1S,3R,4R)-3,4-dimethoxycyclohexyl]-1-methyl-ethyl]-56-hydroxy-46,47-dimethoxy-44-[2-(2-methoxyethoxy)ethoxy]-35,36,37,38,48,49-hexamethyl-64,65-dioxa-57-azatricyclohexatriaconta-24,26,28(48),29(49)-tetraene-50,51,52,53,54-pentone (I-30). The 130 mg mixture was separated by chiral preparative HPLC and then purified by silica gel chromatography (hexane:DCM:EtOAc:MeOH 3:3:1:0 to 3:3:1:0.4) to afford I-31 (45 mg, 35% yield) and I-30 (40 mg, 31% yield) as white solids.

[0566] Chiral separation methods: Column: CHIRALPAK IC Column size: 5.0 cm inner diameter x 25 cm length, 10 μm Sample solution: 1.4mg / mL in mobile phase Injection: 15mL Mobile phase: Hexane / EtOH = 50 / 50 (V / V) Flow rate: 60mL / min Wavelength: UV254nm Temperature: 35℃ I-31: ESI-MS (EI+ ,m / z):1052.1[M+Na] + 。 1 H NMR(400MHz,CDCl3)δ 6.51-5.82(m,4H),5.76-5.03(m,4H),4.51(dd,J=56.0,27.4Hz,1H),4.35-4.06(m,1H),4.00-3.20(m,26H),3.19-2.98(m,5H),2.88-2.48(m,3H),2.40-1.85(m,7H),1.82-1.65(m,11H),1.38(ddd,J=37.8,31.6,21.3Hz,10H),1.21-0.83(m,18H),0.79-0.68(m,1H)。 I-30:ESI-MS(EI + ,m / z):1052.2[M+Na] + 。 1 H NMR(400MHz,CDCl3)δ 6.52-5.81(m,4H),5.77-5.04(m,5H),4.70-4.14(m,2H),4.01-2.97(m,31H),2.64(dd,J=50.7,36.3Hz,3H),2.42-1.68(m,16H),1.50-0.61(m,30H)。

[0567] Example 18: (25E,27E,29E,30E,36R,37S,38R,39R,41S,43S,46S,47R,48R,57R)-46-[(1R)-2-[(1S,3R,4R)-4-dimethylphosphoryloxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-57-hydroxy-47,48-dimethoxy-45-[2-(2-methoxyethoxy)ethoxy]-36,37,38,39,49,50-hexamethyl-66,67-dioxa-5 8-Azatricyclohexatriaconta-25,27,29(49),30(50)-tetraene-51,52,53,54,55-pentone (I-32), (25E,27E,29E,30E,36R,37S,38R,39R,41S,43S,45S,46S,47R,48R,57R)-46-[(1R)-2-[(1S,3R,4R)-4-dimethylphosphoryloxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-57-hydroxy-47,48 -dimethoxy-45-[2-(2-methoxyethoxy)ethoxy]-36,37,38,39,49,50-hexamethyl-66,67-dioxa-58-azatricyclohexatriaconta-25,27,29(49),30(50)-tetraene-51,52,53,54,55-pentone (I-34), and (25E,27E,29E,30E,36R,37S,38R,39R,41S,43S,45R,46S,47R,48R,57R)-46-[(1R Synthesis of 2-[(1S,3R,4R)-4-dimethylphosphoryloxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-57-hydroxy-47,48-dimethoxy-45-[2-(2-methoxyethoxy)ethoxy]-36,37,38,39,49,50-hexamethyl-66,67-dioxa-58-azatricyclohexatriaconta-25,27,29(49),30(50)-tetraene-51,52,53,54,55-pentone (I-33)

[0568] [ka]

[0569] Step 1: (27E,29E,31E,32E,34R,35S,36R,37R,39S,41S,43S,44S,45R,46R,55R)-44-[(1R)-2-[(1S,3R,4R)-4-[tert-butyl(dimethyl)silyl]oxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-45,55-dihydroxy-43,46-dimethoxy-34,35,36,37,47,48-hexamethyl-65,66-dioxa-57-azatricyclohexatriaconta-27,29,31(47),32(48)-tetraene-49,50,51,52,53-pentone. (22E,24E,26E,27E,29R,30S,31R,32R,34S,36S,38S,39S,40R,41R,50R)-40,50-dihydroxy-39-[(1R)-2-[(1S,3R,4R)-4-hydroxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-38,41-dimethoxy-29,30,31,32,42,4 To a solution of 3-hexamethyl-60,61-dioxa-51-azatricyclohexatriaconta-22,24,26(42),27(43)-tetraene-44,45,46,47,48-pentone (2 g, 2.19 mmol), imidazole (0.298 g, 4.38 mmol) and tert-butyl-chloro-dimethyl-silane (0.495 g, 3.28 mmol) were added. The mixture was stirred at 20 °C for 5 h and then poured into ice-cold saturated aqueous NH Cl (10 mL) and Et O:petroleum ether (300 mL, 2:1). The organic layer was washed with saturated aqueous NH Cl (100 mL), water, and brine (100 mL), dried over Na SO , filtered, and concentrated. The residue was purified by silica gel chromatography (10% to 50% EtOAc in petroleum ether) to give the title compound (1.85 g, 82% yield) as a white solid. + , m / z): 1050.2 [M + Na] + . 1H NMR(400MHz,CDCl3)δ 6.44-5.83(m,4H),5.60-5.07(m,4H),4.32-4.04(m,2H),3.79(d,J=13.4Hz,1H),3.70(d,J=6.1Hz,1H),3.65(dd,J=9.8,5.5Hz,1H),3.62-3.53(m,1H),3.43-3.28(m,8H),3.13(s,3H),2.94-2.81(m,1H),2.73(dd,J=16.8,5.9Hz,2H),2.63-2.47(m,1H),2.33(d,J=12.7Hz,2H),2.07-1.89(m,4H),1.89-1.40(m,19H),1.38-1.02(m,15H),1.02-0.76(m,18H),0.69(s,1H),0.05(dd,J=8.2,5.1Hz,6H)。

[0570] Step 2: (27E,29E,31E,32E,38R,39S,40R,41R,43S,45S,48S,49R,50R,59R)-48-[(1R)-2-[(1S,3R,4R)-4-[tert-butyl(dimethyl)silyl]oxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-49,59-dihydroxy-50-methoxy-47-[2-(2-methoxyethoxy)ethoxy]-38,39,40,41,51,52-hexamethyl-69,70-dioxa-61-azatricyclohexatriaconta-27,29,31(51),32(52)-tetraene-53,54,55,56,57-pentone. (27E,29E,31E,32E,34R,35S,36R,37R,39S,41S,43S,44S,45R,46R,55R)-44-[(1R)-2-[(1S,3R,4R)-4-[tert-butyl(dimethyl)silyl]oxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-45,55-dihydroxy-43,46-dimethoxy-34,35,36,37,47,48 in sulfolane (20 mL) To a solution of 1.7 g (1.65 mmol) of 1-hexamethyl-65,66-dioxa-57-azatricyclohexatriaconta-27,29,31(47),32(48)-tetraene-49,50,51,52,53-pentone and 2-(2-methoxyethoxy)ethanol (3.97 g, 33.06 mmol) was added HND-8 (255 mg) at 50 °C under N 2 , and the resulting solution was stirred at 50 °C for 2 h. The reaction mixture was poured into water, extracted with EtOAc, washed with water and brine, dried over Na 2 SO 4 , filtered, and concentrated. The residue was purified by silica gel chromatography (50% EtOAc in petroleum ether) followed by reverse-phase chromatography (85% CH 3 CN in water) to give the title compound (950 mg, 51% yield) as a white solid. ESI-MS(EI + , m / z): 1138.2 [M + Na] + .

[0571] Step 3: (28E,30E,32E,33E,39R,40S,41R,42R,44S,46S,49S,50R,51R,60R)-49-[(1R)-2-[(1S,3R,4R)-4-[tert-butyl(dimethyl)silyl]oxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-60-hydroxy-50,51-dimethoxy-48-[2-(2-methoxyethoxy)ethoxy]-39,40,41,42,52,53-hexamethyl-69,70-dioxa-62-azatricyclohexatriaconta-28,30,32(52),33(53)-tetraene-54,55,56,57,58-pentone. (27E,29E,31E,32E,38R,39S,40R,41R,43S,45S,48S,49R,50R,59R)-48-[(1R)-2-[(1S,3R,4R)-4-[tert-butyl(dimethyl)silyl]oxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-49,59-dihydroxy-50-methoxy-47-[2-(2-methoxyethoxy)ethoxy]-38,39,40,41, To a solution of 51,52-hexamethyl-69,70-dioxa-61-azatricyclohexatriaconta-27,29,31(51),32(52)-tetraene-53,54,55,56,57-pentone (0.5 g, 0.448 mmol), N1,N1,N8,N8-tetramethylnaphthalene-1,8-diamine (1.92 g, 8.96 mmol) and methyl trifluoromethanesulfonate (1.10 g, 6.72 mmol) were added at room temperature. The resulting solution was stirred at 50 °C for 3 h, then filtered and concentrated. The residue was purified by silica gel chromatography (eluting with 0% to 50% EtOAc in petroleum ether) followed by reverse-phase chromatography (0% to 100% CHCN in water) to give the title compound (160 mg, 32% yield) as a pale yellow solid. + , m / z): 1152.2 [M + Na] + .

[0572] Step 4: (23E,25E,27E,28E,34R,35S,36R,37R,39S,41S,44S,45R,46R,55R)-55-hydroxy-44-[(1R)-2-[(1S,3R,4R)-4-hydroxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-45,46-dimethoxy-43-[2-(2-methoxyethoxy)ethoxy]-34,35,36,37,47,48-hexamethyl-64,65-dioxa-56-azatricyclohexatriaconta-23,25,27(47),28(48)-tetraene-49,50,51,52,53-pentone. (28E,30E,32E,33E,39R,40S,41R,42R,44S,46S,49S,50R,51R,60R)-49-[(1R)-2-[(1S,3R,4R)-4-[tert-butyl(dimethyl)silyl]oxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-60-hydroxy-50,51-dimethoxy-48-[2-( A solution of [2-methoxyethoxy)ethoxy]-39,40,41,42,52,53-hexamethyl-69,70-dioxa-62-azatricyclohexatriaconta-28,30,32(52),33(53)-tetraene-54,55,56,57,58-pentone (0.58 g, 0.513 mmol) was added to Py·HF (2.54 g, 25.65 mmol) at 0 °C. The reaction was stirred at room temperature for 3 h, then diluted with DCM and aqueous NaHCO, washed with water and brine, dried over NaSO, filtered, and concentrated. The residue was purified by reverse-phase chromatography (75% CHCN in water) to give the title compound (200 mg, 38% yield) as a white solid. ESI-MS (EI + , m / z): 1038.1 [M + Na] + . 1 H NMR(400MHz,CDCl3)δ 6.49-5.79(m,4H),5.69-5.03(m,4H),4.62(d,J=13.2Hz,1H),4.00-3.07(m,28H),3.02-2. 47(m,6H),2.41-1.68(m,16H),1.54-1.21(m,11H),1.17-0.82(m,18H),0.79-0.55(m,1H).

[0573] Step 5: (25E,27E,29E,30E,36R,37S,38R,39R,41S,43S,46S,47R,48R,57R)-46-[(1R)-2-[(1S,3R,4R)-4-dimethylphosphoryloxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-57-hydroxy-47,48-dimethoxy-45-[2-(2-methoxyethoxy)ethoxy]-36,37,38,39,49,50-hexamethyl-66,67-dioxa-58-azatricyclohexatriaconta-25,27,29(49),30(50)-tetraene-51,52,53,54,55-pentone (I-32). (23E,25E,27E,28E,34R,35S,36R,37R,39S,41S,44S,45R,46R,55R)-55-hydroxy-44-[(1R)-2-[(1S,3R,4R)-4-hydroxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-45,46-dimethoxy-43-[2-(2-methoxyethoxy)ethoxy]-34,35,36,37,47,48-hexamethoxy] in DCM (3 mL) To a solution of 1-64,65-dioxa-56-azatricyclohexatriaconta-23,25,27(47),28(48)-tetraene-49,50,51,52,53-pentone (0.18 g, 0.177 mmol) was added 2,6-di-tert-butyl-4-methylpyridine (0.273 g, 1.33 mmol) and dimethylphosphine chloride (0.1 g, 0.89 mmol, dissolved in 0.5 mL of DCM) at 0 °C. The resulting solution was stirred at 0 °C for 3.5 h, then diluted with EtOAc, washed with saturated aqueous NaHCO, water, and brine, dried over NaSO, filtered, and concentrated. The residue was purified by reverse-phase chromatography (CHCN in water) to afford I-32 (90 mg, 47% yield) as a white solid. ESI-MS(EI + , m / z): 1114.1 [M + Na] + . 1H NMR(500MHz,CDCl3)δ 6.49-5.81(m,4H),5.74-4.96(m,4H),4.67-4.03(m,2H),4.00-3.01(m,29H),2.99-2.46 (m,4H),2.44-1.73(m,17H),1.59-1.22(m,15H),1.19-0.83(m,18H),0.82-0.59(m,1H).

[0574] Step 6: (25E,27E,29E,30E,36R,37S,38R,39R,41S,43S,45S,46S,47R,48R,57R)-46-[(1R)-2-[(1S,3R,4R)-4-dimethylphosphoryloxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-57-hydroxy-47,48-dimethoxy-45-[2-(2-methoxyethoxy)ethoxy]-36,37,38,39,49,50-hexamethyl-66,67-dioxa-58-azatricyclohexatriaconta-25,27,29(49),30(50)-tetraene-51,52,53,54,55-pentone (I-3 4) and (25E,27E,29E,30E,36R,37S,38R,39R,41S,43S,45R,46S,47R,48R,57R)-46-[(1R)-2-[(1S,3R,4R)-4-dimethylphosphoryloxy-3-methoxy-cyclohexyl]-1-methyl-ethyl]-57-hydroxy-47,48-dimethoxy-45-[2-(2-methoxyethoxy)ethoxy]-36,37,38,39,49,50-hexamethyl-66,67-dioxa-58-azatricyclohexatriaconta-25,27,29(49),30(50)-tetraene-51,52,53,54,55-pentone (I-33). 125 mg of the epimeric mixture was separated by chiral preparative HPLC and then purified by silica gel chromatography (hexane:DCM:EtOAc:MeOH 3:3:1:0 to 3:3:1:0.3) to give I-34 (25 mg, 20% yield) and I-33 (15 mg, 12% yield) as white solids.

[0575] Chiral separation methods: Column: CHIRALPAK IC カラムサイズ: 5.0cm inner diameter × 25cm length, 10μm Sample solution: 1.2 mg / mL in mobile phase Injection: 10mL Mobile phase: Hexane / EtOH=40 / 60 (V / V) Flow rate: 25mL / min Wavelength: UV254nm Temperature: 35℃ I-34:ESI-MS(EI + ,m / z):1114.1[M+Na] + . 1 H NMR(500MHz,CDCl3)δ 6.41-6.01(m,3H),5.87(dd,J=83.6,10.7Hz,1H),5.57-5.40(m,1H),5.38-4.97(m,3H),4.57(s,1H),4.02(d,J=20.9Hz,1H),3.92-3.62(m,3 H),3.61-2.94(m,26H),2.78-2.40(m,3H),2.29-1.79(m,9H),1.60-1. 38(m,15H),1.36-1.11(m,9H),1.08-0.76(m,18H),0.75-0.64(m,1H). I-33:ESI-MS(EI + ,m / z):1114.1[M+Na] + . 1 H NMR(500MHz,CDCl3)δ 6.48-5.79(m,4H),5.63-5.02(m,4H),4.56(d,J=62.6Hz,1H),3.99-3.09(m,28H),3.01-2. 49(m,5H),2.40-1.72(m,18H),1.54-1.19(m,14H),1.18-0.81(m,19H),0.78-0.59(m,1H).

[0576] Example 19: (23E,25E,27E,28E,36R,37S,38R,39R,41S,43S,46S,47R,48R,57R)-57-hydroxy-46-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-47,48-dimethoxy-45-[2-(2-methoxyethoxy)ethoxy]-36,37,38,39,49,50-hexamethyl-66,67-dioxa-5 8-Azatricyclohexatriaconta-23,25,27(49),28(50)-tetraene-51,52,53,54,55-pentone (I-35), (23E,25E,27E,28E,36R,37S,38R,39R,41S,43S,45S,46S,47R,48R,57R)-57-hydroxy-46-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-47,4 8-Dimethoxy-45-[2-(2-methoxyethoxy)ethoxy]-36,37,38,39,49,50-hexamethyl-66,67-dioxa-58-azatricyclohexatriaconta-23,25,27(49),28(50)-tetraene-51,52,53,54,55-pentone (I-37), and (23E,25E,27E,28E,36R,37S,38R,39R,41S,43S,45R,46S,47R,48R,57R)-57-hydroxybenzoate Synthesis of -46-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-47,48-dimethoxy-45-[2-(2-methoxyethoxy)ethoxy]-36,37,38,39,49,50-hexamethyl-66,67-dioxa-58-azatricyclohexatriaconta-23,25,27(49),28(50)-tetraene-51,52,53,54,55-pentone (I-36)

[0577] [ka]

[0578] Step 1: (36E,38E,40E,41E,47R,48S,49R,50R,52S,54S,56S,57S,58R,59R,68R)-57-[(1R)-2-[(1S,3R,4R)-4-[2-[tert-butyl(diphenyl)silyl]oxyethoxy]-3-methoxy-cyclohexyl]-1-methyl-ethyl]-68-hydroxy-56,58,59-trimethoxy-47,48,49,50,60,61-hexamethyl-77,78-dioxa-70-azatricyclohexatriaconta-36,38,40(60),41(61)-tetraene-62,63,64,65,66-pentone. (35E,37E,39E,40E,46R,47S,48R,49R,51S,53S,55S,56S,57R,58R,67R)-56-[(1R)-2-[(1S,3R,4R)-4-[2-[tert-butyl(diphenyl)silyl]oxyethoxy]-3-methoxy-cyclohexyl]-1-methyl-ethyl]-57,67-dihydroxy-55,58-dimethoxy-46,47,48,49,59,60-hexamethoxy] in toluene (40 mL) To a suspension of ethyl-77,78-dioxa-69-azatricyclohexatriaconta-35,37,39(59),40(60)-tetraene-61,62,63,64,65-pentone (Intermediate IX prepared according to Example 22, 1.8 g, 1.5 mmol) and 1,8-bis(dimethylamino)naphthalene (6.45 g, 30.08 mmol), methyl trifluoromethanesulfonate (3.70 g, 22.56 mmol) was added dropwise at room temperature under N. After the addition, the mixture was heated to 50 °C for 5 h. Then, the mixture was quenched by adding water (50 mL) and extracted with EtOAc (50 mL) at 0 °C. The organic layer was washed with water (50 mL × 3) and brine (50 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel chromatography (petroleum ether: EtOAc = 3:1) to give the title compound (700 mg, 38% yield) as a pale yellow solid. + , m / z): 1232.2 [M + Na] + . 1H NMR(400MHz,CDCl3)δ 7.70-7.68(m,4H),7.43-7.26(m,6H),6.40-5.87(m,4H),5.68-5.07(m,4H),4.67(s,1H),4.48-4.13(m,1H),3.81-3.57(m,7H),3.47-3.33(m,5H),3.20-3.08(m,7H),3.07-2.97(m,1H),2.71-2.50(m,2H),2.35-2.20(m,2H),2.09-1.97(m,3H),1.70-1.66(m,6H),1.61-1.58(m,11H),1.38-1.20(m,10H),1.15-1.10(m,5H),1.09-1.05(m,10H),0.98-0.73(m,13H),0.71-0.66(m,1H)。

[0579] Step 2: (23E,25E,27E,28E,32R,33S,34R,35R,37S,39S,41S,42S,43R,44R,53R)-53-hydroxy-42-[(1R)-2-[(1S,3R,4R)-4-(2-hydroxyethoxy)-3-methoxy-cyclohexyl]-1-methyl-ethyl]-41,43,44-trimethoxy-32,33,34,35,45,46-hexamethyl-62,63-dioxa-54-azatricyclohexatriaconta-23,25,27(45),28(46)-tetraene-47,48,49,50,51-pentone. (36E,38E,40E,41E,47R,48S,49R,50R,52S,54S,56S,57S,58R,59R,68R)-57-[(1R)-2-[(1S,3R,4R)-4-[2-[tert-butyl(diphenyl)silyl]oxyethoxy]-3-methoxy-cyclohexyl]-1-methyl-ethyl]-68-hydroxy-56 To a solution of 58,59-trimethoxy-47,48,49,50,60,...

Claims

1. A compound of formula I-a-1, formula I-a-2, or formula I-b-1, or a pharma- ceutically acceptable salt thereof: 【Chemistry 1】 [In the formula, X is -CR ２ -, -NRCO-, -NRCO ２ -, -NRCONR-, -NRSO ２ - or -SO ２ NR-; X 3 is a covalent bond, -CR ２ -, -NR-, -NRCO-, -NRCO ２ -, -NRCONR-, -NRSO ２ -, -O-, -S-, or -SO ２ NR-; L １ is a covalent bond or C １～３０ A divalent linear or branched saturated or unsaturated hydrocarbon chain, wherein 1 to 10 methylene units of said chain are independently selected from the group consisting of -Cy １ -, -O-, -S-, -SO ２ -, -C(O)-, -C(S)-, -CR ２ -, -CF ２ -, -P(O)(R)-, -SiR ２ substituted with -, -Si(OR)(R)-, or -NR-; Each -Cy １ - is an optionally substituted bivalent ring which is independently phenylene, a 4- to 7-membered saturated or partially unsaturated heterocyclylene having 1-2 heteroatoms which are independently nitrogen, oxygen, or sulfur, or a 5- to 6-membered heteroarylene having 1-4 heteroatoms which are independently nitrogen, oxygen, or sulfur; Each R is independently hydrogen or C １～６ an optionally substituted group which is an aliphatic, 3- to 8-membered saturated or partially unsaturated monocyclic carbocycle, phenyl, a 4- to 7-membered saturated or partially unsaturated heterocycle having 1-2 heteroatoms which are independently nitrogen, oxygen, or sulfur, or a 5- to 6-membered heteroaryl ring having 1-4 heteroatoms which are independently nitrogen, oxygen, or sulfur; two R groups on the same atom, taken together with their intervening atoms, form a 4-7 membered saturated, partially unsaturated, or aryl ring having, in addition to the same atom to which they are attached, 0-3 heteroatoms which are independently nitrogen, oxygen, or sulfur; R １ and R ２ are independently methyl, —CHF 2 , 【Chemistry 2】 and R ３ is -OR, where R is H or C 1-4 alkyl; R ３’ is hydrogen; R ４ is -OR, where R is C 1-4 alkyl; R 6 is -OR, where R is C 1-4 alkyl optionally substituted with halogen -NR 2 ; R ５ and R ５’ each taken together forms =O]; 【Chemistry 3】 [In the formula, X is a covalent bond, -CR 2 -, -NR-, -NRCO-, -NRCO 2 -, -NRCONR-, -NRSO 2 -, -O-, -S- or -SO 2 NR-; X 3 is —CR 2 —, —NRCO—, —NRCO 2 —, —NRCONR—, —NRSO 2 —, or —SO 2 NR—; L 1 is a covalent bond or a C 1-30 divalent linear or branched saturated or unsaturated hydrocarbon chain, wherein 1 to 10 methylene units of said chain are independently and optionally replaced with -Cy 1 -, -O-, -S-, -SO 2 -, -C(O)-, -C(S)-, -CR 2 -, -CF 2 -, -P(O)(R)-, -SiR 2 -, -Si(OR)(R)-, or -NR-; each -Cy 1 - is independently an optionally substituted bivalent ring which is phenylene, a 4- to 7-membered saturated or partially unsaturated heterocyclylene having 1 to 2 heteroatoms which are independently nitrogen, oxygen, or sulfur, or a 5- to 6-membered heteroarylene having 1 to 4 heteroatoms which are independently nitrogen, oxygen, or sulfur; each R is independently hydrogen or an optionally substituted group which is a C 1-6 aliphatic, 3- to 8-membered saturated or partially unsaturated monocyclic carbocycle, phenyl, 4- to 7-membered saturated or partially unsaturated heterocycle having 1-2 heteroatoms which are independently nitrogen, oxygen, or sulfur, or a 5- to 6-membered heteroaryl ring having 1-4 heteroatoms which are independently nitrogen, oxygen, or sulfur; two R groups on the same atom, taken together with their intervening atoms, form a 4-7 membered saturated, partially unsaturated, or aryl ring having, in addition to the same atom to which they are attached, 0-3 heteroatoms which are independently nitrogen, oxygen, or sulfur; R 1 and R 2 are independently methyl, —CHF 2 , 【Chemistry 4】 and R 3 is —OR, where R is H or C 1-4 alkyl; R 3′ is hydrogen; R 4 is —OR, where R is C 1-4 alkyl; R 6 is -OR, where R is C 1-4 alkyl optionally substituted with halogen -NR 2 ; R 5 and R 5' each together form =O; 【Chemistry 5】 [In the formula, X is a covalent bond, -CR 2 -, -NR-, -NRCO-, -NRCO 2 -, -NRCONR-, -NRSO 2 -, -O-, -S- or -SO 2 NR-; L 1 is a covalent bond or a C 1-30 divalent linear or branched saturated or unsaturated hydrocarbon chain, wherein 1 to 10 methylene units of said chain are independently and optionally replaced with -Cy 1 -, -O-, -S-, -SO 2 -, -C(O)-, -C(S)-, -CR 2 -, -CF 2 -, -P(O)(R)-, -SiR 2 -, -Si(OR)(R)-, or -NR-; each -Cy 1 - is independently an optionally substituted bivalent ring which is phenylene, a 4- to 7-membered saturated or partially unsaturated heterocyclylene having 1 to 2 heteroatoms which are independently nitrogen, oxygen, or sulfur, or a 5- to 6-membered heteroarylene having 1 to 4 heteroatoms which are independently nitrogen, oxygen, or sulfur; each R is independently hydrogen or an optionally substituted group which is a C 1-6 aliphatic, 3- to 8-membered saturated or partially unsaturated monocyclic carbocycle, phenyl, 4- to 7-membered saturated or partially unsaturated heterocycle having 1-2 heteroatoms which are independently nitrogen, oxygen, or sulfur, or a 5- to 6-membered heteroaryl ring having 1-4 heteroatoms which are independently nitrogen, oxygen, or sulfur; two R groups on the same atom, taken together with their intervening atoms, form a 4-7 membered saturated, partially unsaturated, or aryl ring having, in addition to the same atom to which they are attached, 0-3 heteroatoms which are independently nitrogen, oxygen, or sulfur; R 1 is methyl, —CHF 2 , 【Chemistry 6】 and R 3 is —OR, where R is H or C 1-4 alkyl; R 3′ is hydrogen; R 4 is —OR, where R is C 1-4 alkyl; R 6 is -OR, where R is C 1-4 alkyl optionally substituted with halogen -NR 2 ; R 5 and R 5' each together form =O.

2. L in the compound of formula I-a-1, formula I-a-2 or formula I-b-1 １ But, C １～１０ a divalent linear or branched saturated or unsaturated hydrocarbon chain, wherein 1 to 5 methylene units of said chain are independently and optionally -Cy １ -, -O-, -S-, -SO ２ -, -C(O)-, -C(S)-, -CR ２ -, -CF ２ 2. The compound of claim 1, or a pharma- ceutically acceptable salt thereof, which is substituted with -, -P(O)(R)-, or -NR-.

3. L in the compound of formula I-a-1, formula I-a-2 or formula I-b-1 １ But -CH ２ --, --CH ２ CH ２ -, -(CH ２ ) ３ -, -(CH ２ ) ４ -, -(CH ２ ) ５ --, --CH ２ CH ２ O-, -(CH ２ CH ２ O) ２ -, -(CH ２ CH ２ O) ３ --, --CH ２ CH ２ OCH ２ CH ２ --, --CH ２ CH ２ SO ２ CH ２ CH ２ O- or -CH ２ CH ２ OCH ２ CH ２ OCH ２ CH ２ 2. The compound of claim 1, wherein R is - or a pharma- ceutically acceptable salt thereof.

4. L in the compound of formula I-a-1, formula I-a-2 or formula I-b-1 １ The compound of claim 1 , or a pharma- ceutically acceptable salt thereof, wherein is a covalent bond.

5. In the compound of formula I-a-1, formula I-a-2 or formula I-b-1, each R is independently hydrogen, or C １～６ 2. The compound of claim 1, or a pharma- ceutically acceptable salt thereof, which is an optionally substituted group that is an aliphatic, a 3-8 membered saturated or partially unsaturated monocyclic carbocycle, a phenyl, a 4-7 membered saturated or partially unsaturated heterocycle having 1-2 heteroatoms which are independently nitrogen, oxygen, or sulfur, or a 5-6 membered heteroaryl ring having 1-4 heteroatoms which are independently nitrogen, oxygen, or sulfur.

6. 13. A pharma- ceutically acceptable composition comprising the compound of claim 1 or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable carrier, adjuvant, or vehicle.

7. The compound according to claim 1, wherein the compound represented by formula I-a-1 or formula I-a-2 is represented by formula I-b-2, or a pharma- ceutically acceptable salt thereof. 【Chemistry 7】 8. The compound according to claim 1, wherein the compound represented by formula I-a-1 or formula I-a-2 is represented by formula I-b-3, or a pharma- ceutically acceptable salt thereof. 【Chemistry 8】 9. The compound of claim 1, or a pharma- ceutically acceptable salt thereof, which is a compound of formula I-a-1, or a pharma- ceutically acceptable salt thereof:

10. The compound of claim 1, or a pharma- ceutically acceptable salt thereof, which is a compound of formula I-a-2, or a pharma- ceutically acceptable salt thereof:

11. The compound of claim 1, or a pharma- ceutically acceptable salt thereof, which is a compound of formula I-b-1, or a pharma- ceutically acceptable salt thereof:

12. The compound: 【Chemistry 9-1】 【Chemistry 9-2】 【Chemistry 9-3】 【Chemistry 9-4】 【Chemistry 9-5】 【Chemistry 9-6】 【Chemistry 9-7】 【Chemistry 9-8】 【Chemistry 9-9】 【Chemistry 9-10】 【9-11】 【Chemistry 9-12】 【Chemistry 9-13】 【Chemistry 9-14】 【Chemistry 9-15】 【Chemistry 9-16】 【Chemistry 9-17】 【Chemistry 9-18】

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