Selective rapamycin analogs and uses thereof

Abstract

Disclosed are compounds of Formula (I) and pharmaceutical compositions thereof for the treatment of disorders or diseases mediated by the mTOR pathway by more selective binding to FKBP12 in the FK506 binding protein (FKBP) group. Those compounds are useful for the treatment of age or aging related diseases, diabetes, cancer, and inflammation related disorders due to unusual pharmacokinetic profiles and enhanced pharmacodynamic selectivity in target FKBP.

Description

[0001] Cross-references to related applications

[0002] This application claims priority to U.S. Provisional Application No. 63 / 494,839, filed April 7, 2023; U.S. Provisional Application No. 63 / 515,184, filed July 24, 2023; and U.S. Provisional Application No. 63 / 624,875, filed January 25, 2024, all of which are hereby incorporated in their entirety by reference. Technical Field

[0003] This article provides FKBP12-selective rapamycin analogues, pharmaceutical compositions comprising said analogues, and methods for treating diseases, disorders, and conditions, said methods comprising administering said analogues or pharmaceutical compositions thereof. Background of the Invention

[0005] Rapamycin is a known macrolide antibiotic produced by *Streptomyces hygroscopicus*, see, for example, McAlpine, JB et al., *J. Antibiotics* (1991) 44:688; Schreiber, SL et al., *J. Am. Chem. Soc.* (1991) 113:7433; and U.S. Patent No. 3,929,992. The following numbering conventions for rapamycin and its derivatives as used herein are shown below:

[0006]

[0007] Rapamycin is a potent immunosuppressant used to prevent organ transplant rejection and to treat certain types of cancer. It has also been shown to be useful for the prevention or treatment of systemic lupus erythematosus, insulin-dependent diabetes mellitus, skin disorders such as psoriasis, smooth muscle cell proliferation and endothelial thickening following vascular injury, adult T-cell leukemia / lymphoma, malignancies, inflammatory cardiac diseases, anemia, and increased neurite growth. Furthermore, rapamycin analogues (so-called “rapalog”) have been shown to be effective against liver fibrosis. See, for example, Liver Int. (2014) 34(10):1513-21.

[0008] In eukaryotic cells, rapamycin and its analogues (rapalog) inhibit TOR (target of rapamycin) signaling. In mammalian cells, mTOR (mammalian target of rapamycin) exists as two distinct multiprotein complexes, described as the mTORC1 and mTORC2 complexes, both of which sense nutrient and energy availability and integrate inputs from growth factors and stress signaling. mTORC1 integrates signals from growth factors and nutrients and controls cell growth and metabolism. Laplante M. et al. Cell. (2012) 149(2):274-93. mTORC1 is a key regulator of protein translation and autophagy.

[0009] In animal models, rapamycin analogues extended lifespan and delayed the onset of age-related diseases. Aging, like other biological processes, is regulated by signaling pathways such as the TOR pathway (named "TOR" in this case to include the yeast and *C. elegans* systems) and the mTORC1 pathway in mammals. Regulation of TOR and mTORC1 signaling extends lifespan and delays the onset of age-related diseases in a wide range of organisms from flies to mammals. For example, inhibiting the TOR pathway through gene mutations has prolonged the lifespan of yeast, *C. elegans*, and *Drosophila*, and inhibiting the mTORC1 pathway has prolonged the lifespan of mice (Kaeberlein et al., Science (2005) 310:1193-1196; Kapahi et al., Curr Biol (2004) 14:885-890; Selman et al., Science (2009) 326:140-144; Velai et al., Nature (2003) 426:620). Furthermore, the mTORC1 inhibitor rapamycin has prolonged the lifespan of mice even when administered late in life (Harrison et al., Nature (2009) 460(7253):392-395). These data raise the possibility that drugs targeting the mammalian TOR (mTOR) pathway will have therapeutic effects on aging and age-related diseases in humans (M. Leslie, Science, 2013, 342). For example, J. Mannick et al. described in SciTransl Med. (2014) 6(268):268ra179 that mTOR inhibition improves immune function in older adults.

[0010] Mitochondrial myopathy (MM) is the most common manifestation of adult-onset mitochondrial disease and exhibits multifaceted tissue-specific stress responses: (1) transcriptional responses, including metabolic cytokines FGF21 and GDF15; (2) remodeling of one-carbon metabolism; and (3) mitochondrial unfolded protein responses (Khan et al., Cell Metabolism 26, 419-428, August 1, 2017). Rapamycin-induced mTORC1 inhibition downregulated all components of ISRmt (integrated mitochondrial stress responses), improved all MM markers, and reversed the progression of even late-stage MM without inducing mitochondrial biogenesis. Therefore, rapamycin and rapamycin analogues are considered to have potential value in addressing many unmet clinical needs.

[0011] Epilepsy caused by mutations in the Tsc1 / Tsc2 complex can be treated with rapamycin or rapamycin analogues. (Zeng et al., Ann. Neurology, April 2008; 63(4)444-453). It is advantageous to have rapamycin analogues that can suppress epilepsy without interfering with other tissues expressing mTORC1—therefore, in this context, more selective mTORC1 inhibitors would be desirable.

[0012] mTORC1 is a key regulator of protein translation and autophagy. The mTORC1 complex is sensitive to allosteric mTOR inhibitors such as rapamycin and rapamycin analogs. The mode of action of rapamycin and previously developed rapamycin analogs involves the formation of an intracellular complex with an FK506-binding protein, which may include FKBP12, FKBP12.6, FKBP13, FKBP25, FKBP51, or FKBP52 (these six FKBPs will be referred to as “FKBPs” or “FKBPs” in this document), followed by the binding of the FKBP-rapamycin analog complex to the FRB (FK506-rapamycin-binding) domain of mTOR (Marz A.M. et al. Mol Cell Biol. (2013) 33(7):1357-1367). This interaction between the FKBP-rapamycin analog complex and mTORC1 results in allosteric inhibition of the complex. Rapamycin and its analogues, such as RAD001 (everolimus), have gained clinical relevance by inhibiting the activity of mTORC1, which is associated with both benign and malignant proliferative disorders (Royce ME et al., Breast Cancer (Auckl). (2015) 9:73-79; Pleniceanu O. et al., Kidney Int Rep. (2018) 3(1):155-159). Various FKBPs are expressed to varying degrees in a wide range of cell types and organs in vivo.

[0013] Rapamycin treatment has also been reported to selectively target CD40-mediated B cell proliferation and differentiation (Atsuko Sakata et al., Immunology Letters. Vol. 68, No. 2-3, June 1, 1999, pp. 301-309). CD40 is a cell surface receptor and part of the tumor necrosis factor (TNF) receptor superfamily. CD40 is expressed on antigen-presenting cells such as B cells, macrophages, and dendritic cells, as well as some non-immune cells and tumors (Dakal et al., Immunobiology 2020, 225:151899). Activation of resting B cells requires initial triggering by the B cell antigen receptor (BCR) and secondary stimulation via various cytokine receptors and B cell activating molecules, including CD40.

[0014] The interaction between CD40 and its ligand CD40L provides a co-stimulatory signal that is essential for the survival of many cell types and is required for immune responses such as germinal center formation, antibody responses to T-dependent antigens, and “permitting” dendritic cells to mature and become effective in triggering T cell activation and differentiation (see, for example, Kawabe et al., Immunity 1994, 1:167-178; Elgueta et al., Immunol. Rev. 2009, 229:152-172).

[0015] CD40-CD40L signaling is involved in autoimmune conditions primarily driven by autoantibodies, such as systemic rheumatic diseases in which autoantibodies play a significant role in disease progression (e.g., multiple sclerosis, autoimmune nephritis, rheumatoid arthritis, Sjögren's syndrome, and systemic lupus erythematosus), as well as non-rheumatic conditions with autoantibody components (e.g., myasthenia gravis, Grave's disease, and neuromyelitis optica) (see, Karnell et al., Adv Drug Delivery Rev. 2019, 141:92-103). Furthermore, because CD40-CD40L signaling is essential for the activation of antigen-presenting cells, blocking antigen presentation by dendritic cells or B cells can affect CD8+ T cell responses in some diseases (e.g., Denic et al., Expert Opin Ther Targets 2013, 17:1053-1066). Altered CD40-CD40L signaling is also associated with other diseases and conditions such as cardiovascular disease and transplantation (see, for example, Dakal et al., Immunobiology 2020, 225:151899; Pamukcu et al., Ann. Med. 2011, 43:331; 340; Pinelli et al., Immunotherapy 2015, 7:399-410).

[0016] For example, CD40 co-stimulatory blockade combined with immunosuppression by rapamycin provided long-term islet and kidney allogeneic graft survival (90 days, 94 days, >120 days, >120 days, and >120 days), with only one recipient showing evidence of allogeneic graft rejection. The CD40 / rapamycin regimen was also tested in four individual kidney transplant recipients. All four recipients achieved long-term kidney allogeneic graft survival (100% at day 120), which was superior to kidney allogeneic graft survival using a triple immunosuppressive regimen (tacrolimus, mycophenolate mofetil, and steroids) (62.9% at day 120) (T. Oura 1, K. Hotta et al., American Journal of Transplantation. Vol. 17, No. 3, March 2017, pp. 646-656).

[0017] Because different FKBPs are expressed differentially in different tissues, and because it is necessary to avoid mTORC1 inhibition in specific cell types in some cases, rapamycin analogs selective for individual FKBPs will be needed to enhance the ability to target specific tissues. For example, rapamycin analogs selective for FKBP12 can target tissues with high FKBP12 expression but not tissues with low (or no) FKBP12 levels. Selective FKBP analogs will provide patients with unique and clinically beneficial advantages because they avoid adverse events caused by widespread mTORC1 inhibition.

[0018] There is also a need for rapamycin analogues that can be administered in combination with anti-CD40 antibodies to treat diseases in which autoantibodies play an important role in disease progression, to induce an immune response and / or prevent organ transplant rejection.

[0019] There is also a need for rapamycin analogues that can be combined with immunotherapeutic agents that target T cells to specific tumors, so that the rapamycin analogues do not inhibit or interfere with the immunotherapy. Invention Overview

[0021] In one aspect, this paper provides compounds of formula I:

[0022]

[0023] Or its pharmaceutically acceptable salt;

[0024] Where R 1 It is hydrogen, C 1-6 Alkyl, heterocyclic, aryl, heteroaryl, -C 0-6 Alkylene-SO2R 4 or -C 0-6 Alkylene-SO2R 5 ; where C 1-6 Alkyl, heterocyclic, aryl, and heteroaryl groups are optionally separated by one or two R groups. 1a Group substitution;

[0025] R 2 It is a heterocyclic group, aryl group, heteroaryl group, -C 0-6 Alkylene-SO2R 4 or -C 0-6 Alkylene-SO2R 5 ; wherein the heterocyclic group, aryl group, and heteroaryl group are optionally surrounded by one or two R groups. 2a Group substitution;

[0026] Or R 1 and R 2 Together with the attached nitrogen, it forms an optional structure with one or two R 1aN-linked heteroaryl groups or N-linked heterocyclic groups with substituent groups;

[0027] R 3 Choose from the following groups: - OR a 3-6 membered heterocyclic groups and 3-6 membered heteroaryl groups;

[0028] R a Choose from the following groups: H, -P(O)(R b )2、-C(O)R c -C(O)OR c C 1-6 Alkyl and C 1-6 Hydroxyalkyl;

[0029] Each R b Choose independently from the following groups: H and C 1-6 alkyl;

[0030] R c Choose from the following groups: H, C 1-6 Alkyl and C 1-6 Hydroxyalkyl;

[0031] R 4 It is C 2-6 Alkyl, C 3-6 cycloalkyl, C 1-6 hydroxyalkyl, heterocyclic, aryl, or heteroaryl; wherein the heterocyclic, aryl, and heteroaryl groups are optionally surrounded by one or two R groups. 4a Group substitution; and

[0032] R 5 It is a heterocyclic, aryl, or heteroaryl group; wherein the heterocyclic, aryl, or heteroaryl group is optionally surrounded by one or two R groups. 5a Group substitution;

[0033] One or two R 1a R 2a R 4a and R 5a Each of the groups, when present, is independently selected from C. 1-6 Alkyl, C 1-6 Hydroxyalkyl, hydroxyl, halogen, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 Haloalkoxy and cyano; or any two R 1a R 2a R 4a and R 5a When groups exist on the same carbon atom, they form oxo groups together.

[0034] Where R 1When it is hydrogen, R 2 Not -C 0-6 Alkylene-SO2R 4 .

[0035] In one embodiment, the compound of formula (I) is the compound of formula (Ia):

[0036]

[0037] Or its pharmaceutically acceptable salt;

[0038] Where R 1 Is it hydrogen or C? 1-6 alkyl;

[0039] R 2 It is optional to be controlled by one or two Rs 2a Heterocyclic, aryl, or heteroaryl groups substituted with functional groups;

[0040] R 3 Choose from the following groups: - OR a 3-6 membered heterocyclic groups and 3-6 membered heteroaryl groups;

[0041] R a Choose from the following groups: H, -P(O)(R b )2、-C(O)R c -C(O)OR c C 1-6 Alkyl and C 1-6 Hydroxyalkyl;

[0042] Each R b Choose independently from the following groups: H and C 1-6 alkyl;

[0043] R c Choose from the following groups: H, C 1-6 Alkyl and C 1-6 Hydroxyalkyl; and

[0044] One or two R 2a Each of the groups, when present, is independently selected from C. 1-6 Alkyl, C 1-6 Hydroxyalkyl, hydroxyl, halogen, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 Haloalkoxy and cyano; or any two R 2a When groups exist on the same carbon atom, they form oxo groups together.

[0045] In one embodiment, the compound of formula (I) is the compound of formula (Ib):

[0046]

[0047] Or its pharmaceutically acceptable salt;

[0048] Where R 1 and R 2 Together with the attached nitrogen, it forms an optional structure with one or two R 1a N-linked heteroaryl groups or N-linked heterocyclic groups with substituent groups;

[0049] R 3 Choose from the following groups: - OR a 3-6 membered heterocyclic groups and 3-6 membered heteroaryl groups;

[0050] R a Choose from the following groups: H, -P(O)(R b )2、-C(O)R c -C(O)OR c C 1-6 Alkyl and C 1-6 Hydroxyalkyl;

[0051] Each R b Choose independently from the following groups: H and C 1-6 alkyl;

[0052] R c Choose from the following groups: H, C 1-6 Alkyl and C 1-6 Hydroxyalkyl;

[0053] One or two R 1a Each of the groups, when present, is independently selected from C. 1-6 Alkyl, C 1-6 Hydroxyalkyl, hydroxyl, halogen, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy and cyano groups; or two R groups 1a When groups exist on the same carbon atom, they form oxo groups together.

[0054] In one embodiment, the compound of formula (I) is the compound of formula (Ic):

[0055]

[0056] Or its pharmaceutically acceptable salt;

[0057] Where R 1 Is it hydrogen or C? 1-6 alkyl;

[0058] R2 It is -C 0-6 Alkylene-SO2R 4 or -C 0-6 Alkylene-SO2R 5 ;

[0059] R 3 Choose from the following groups: - OR a 3-6 membered heterocyclic groups and 3-6 membered heteroaryl groups;

[0060] R a Choose from the following groups: H, -P(O)(R b )2、-C(O)R c -C(O)OR c C 1-6 Alkyl and C 1-6 Hydroxyalkyl;

[0061] Each R b Choose independently from the following groups: H and C 1-6 alkyl;

[0062] R c Choose from the following groups: H, C 1-6 Alkyl and C 1-6 Hydroxyalkyl;

[0063] R 4 It is C 2-6 Alkyl, C 3-6 cycloalkyl, C 1-6 hydroxyalkyl, heterocyclic, aryl, or heteroaryl; wherein the heterocyclic, aryl, and heteroaryl groups are optionally surrounded by one or two R groups. 4a Group substitution;

[0064] R 5 It is a heterocyclic, aryl, or heteroaryl group; wherein the heterocyclic, aryl, or heteroaryl group is optionally surrounded by one or two R groups. 5a Group substitution;

[0065] One or two R 4a and R 5a Each of the groups, when present, is independently selected from C. 1-6 Alkyl, C 1-6 Hydroxyalkyl, hydroxyl, halogen, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 Haloalkoxy and cyano; or any two R 4a and R 5a When groups exist on the same carbon atom, they form oxo groups together.

[0066] Where R 1When it is hydrogen, R 2 Not -C 0-6 Alkylene-SO2R 4 .

[0067] This disclosure provides at least the following implementation methods:

[0068] a) Compounds of formula (PI), formula (P-Ia), formula (P-Ib), formula (P-Ic), formula (P-2), formula (I), formula (Ia), formula (Ib), formula (Ic), or pharmaceutically acceptable salts thereof;

[0069] b) Compounds selected from compounds 3-30, 43-65, or 67-74, or pharmaceutically acceptable salts thereof;

[0070] c)(a) or (b) of the compounds, wherein the compounds selectively bind FKBP12 more than other FK506-binding proteins compared to everolimus;

[0071] d) A pharmaceutical composition comprising a compound of formula (PI), formula (P-Ia), formula (P-Ib), formula (P-Ic), formula (P-2), formula (I), formula (Ia), formula (Ib), formula (Ic), or formula (Ic) in a pharmaceutically acceptable excipient, diluent, or carrier, or a pharmaceutically acceptable salt thereof.

[0072] e) A pharmaceutical composition comprising a compound of (b) or (c) in a pharmaceutically acceptable excipient, diluent, or carrier;

[0073] f) A pharmaceutical composition comprising the compounds of (a)-(c) in a pharmaceutically acceptable excipient, diluent or carrier and at least one additional therapeutically active agent;

[0074] g) Combination therapy, wherein the combination therapy comprises a therapeutically effective amount of the compound of (a)-(c) or the pharmaceutical composition of (d)-(f) and an anti-CD40 antibody;

[0075] h) A method for treating a subject with a corresponding need for a disease or disorder mediated by the mTOR pathway, the method comprising administering a therapeutically effective amount of the compound of (a)-(c), the pharmaceutical composition of (d)-(f), or the combination therapy of (g).

[0076] The method i)(h) wherein the target tissue, target organ or target cell associated with the pathology of the disease or disorder has an FKBP12 level sufficient to inhibit mTORC1;

[0077] Methods j)(h) or (i), wherein a therapeutically effective amount of the compound of (a)-(c) or the pharmaceutical composition of (d)-(f) has a sufficiently high affinity for FKBP12 to inhibit mTORC1;

[0078] The method of k)(h)-(j), wherein the disease or disorder is selected from sarcopenia, skin atrophy, cherry angioma, seborrheic keratosis, cerebral atrophy, atherosclerosis, arteriosclerosis, emphysema, osteoporosis, osteoarthritis, hypertension, erectile dysfunction, cataract, macular degeneration, glaucoma, stroke, cerebrovascular disease (stroke), chronic kidney disease, diabetic nephropathy, impaired liver function, liver fibrosis, autoimmune hepatitis, endometrial hyperplasia, metabolic dysfunction, renal vascular disease, hearing loss, activity deficit, cognitive decline, tendon stiffness, and heart failure. It can impair conditions such as cardiac hypertrophy and / or systolic dysfunction and / or diastolic dysfunction and / or hypertension, cardiac dysfunction leading to decreased ejection fraction, immunosenescence, Parkinson's disease, Alzheimer's disease, cancer, immunosenescence leading to cancer due to decreased immune surveillance, infection due to decreased immune function, chronic obstructive pulmonary disease (COPD), obesity, loss of taste, loss of smell, arthritis, cancer in which tumors have elevated levels of mTORC1 signaling and / or sufficient levels of FKBP12 to allow inhibition of mTORC1, and type II diabetes.

[0079] The method of l)(h)-(j), wherein the disease or disorder is selected from the following age-related disorders or diseases: sarcopenia; skin atrophy; cherry angioma; seborrheic keratosis; cerebral atrophy; atherosclerosis; arteriosclerosis; emphysema; osteoporosis; osteoarthritis; hypertension; erectile dysfunction; cataract; macular degeneration; glaucoma; stroke; cerebrovascular disease (stroke); chronic kidney disease; diabetic nephropathy; impaired liver function; liver fibrosis; autoimmune hepatitis; endometrial hyperplasia; metabolic dysfunction; renal vascular disease; hearing loss; activity deficit; cognitive decline; muscle Tendon stiffness; cardiac dysfunction such as cardiac hypertrophy and / or systolic and / or diastolic dysfunction and / or hypertension; cardiac dysfunction leading to decreased ejection fraction; immunosenescence; Parkinson's disease; Alzheimer's disease; cancer; immunosenescence leading to cancer due to decreased immune surveillance; infections due to decreased immune function; chronic obstructive pulmonary disease (COPD); obesity; anosmia; anosmia; arthritis; cancers in which tumors have elevated levels of mTORC1 signaling and / or sufficient levels of FKBP12 to allow inhibition of mTORC1; and type II diabetes.

[0080] The method of m)(h)-(j), wherein the disease or disorder is cancer;

[0081] The method of n)(m), wherein the cancer is selected from renal cell carcinoma, renal cell carcinoma, colorectal cancer, uterine sarcoma, endometrial cancer, endometrial cancer, breast cancer, ovarian cancer, cervical cancer, gastric cancer, fibrosarcoma, pancreatic cancer, liver cancer, melanoma, leukemia, multiple myeloma, nasopharyngeal carcinoma, prostate cancer, lung cancer, glioblastoma, bladder cancer, mesothelioma, head cancer, rhabdomyosarcoma, sarcoma, lymphoma and cervical cancer;

[0082] The method of o)(k), wherein the disease is Alzheimer's disease,

[0083] The method of p)(h), wherein the disease is graft-versus-host disease (GvHD).

[0084] The method of q)(h), wherein the disease is facial angiofibroma associated with tuberous sclerosis syndrome.

[0085] The method of r)(h), wherein the disease is advanced unresectable or metastatic malignant perivascular epithelioid cell tumor;

[0086] s) A method for inducing immune tolerance and / or preventing transplant rejection in subjects with appropriate need, the method comprising administering a therapeutically effective amount of the compound of (a)-(c), the pharmaceutical composition of (d)-(f), or the combination therapy of (g).

[0087] t) The method according to any one of claims (h)-(s), wherein the compound is selected from the group consisting of compounds 16-19, 26-28, 45, 50, 63, 68-72 and 74.

[0088] The compounds of (a)-(c), the pharmaceutical compositions of (d)-(f), or the combination therapy of (g) are used for the treatment of diseases or disorders;

[0089] v)(a)-(c) compounds, (d)-(f) pharmaceutical compositions or (g) combination therapies, for use in treating mTOR pathway-mediated diseases or disorders in subjects with corresponding needs;

[0090] w)(u)-(v) compounds, wherein the compounds are selected from compounds 16-19, 26-28, 45, 50, 63, 68-72 and 74;

[0091] x) Use of therapeutically effective amounts of compounds (a)-(c), pharmaceutical compositions (d)-(f), or combination therapies (g) in the manufacture of a medicament for the treatment of a disease or disorder;

[0092] y) the use of therapeutically effective amounts of compounds (a)-(c), pharmaceutical compositions (d)-(f), or combination therapies (g) in the manufacture of a medicament for the treatment of a subject with an mTOR-mediated disease or disorder; and

[0093] Use of z)(x)-(y), wherein the compound is selected from compounds 16-19, 26-28, 45, 50, 63, 68-72 and 74.

[0094] In some embodiments, the compounds and pharmaceutical compositions disclosed herein for use in treating disorders or diseases mediated by the mTOR pathway are more selective in binding FKBP12 than other proteins in the FK506 binding protein (FKBP) group.

[0095] In some other embodiments, the compounds and pharmaceutical compositions disclosed herein for use in the treatment of disorders or diseases mediated by the mTOR pathway exhibit unusual and surprising pharmacokinetic profiles and enhanced pharmacodynamic selectivity in the target FKBP. These compounds and pharmaceutical compositions may be used to treat age-related diseases, diabetes-related complications, cancer, and inflammation-related disorders.

[0096] In some other embodiments, the compounds and pharmaceutical compositions disclosed herein for use in the treatment of disorders or diseases mediated by the mTOR pathway, wherein the compounds inhibit S6K1 phosphorylation in FKBP12 KO cells at least twice as efficiently as the rapamycin analog RAD001 compared to cells expressing FKBP12.

[0097] In some other embodiments, the compounds and pharmaceutical compositions disclosed herein for use in the treatment of disorders or diseases mediated by the mTOR pathway, wherein the compounds inhibit S6K1 phosphorylation in FKBP12 KO cells at least ten times less efficiently than the rapamycin analog RAD001 compared to cells expressing FKBP12.

[0098] In some other embodiments, the compounds and pharmaceutical compositions disclosed herein for use in the treatment of disorders or diseases mediated by the mTOR pathway, wherein the compounds inhibit S6K1 phosphorylation in FKBP12 KO cells at least 100-fold less efficiently than the rapamycin analog RAD001 compared to cells expressing FKBP12.

[0099] In some other embodiments, the compounds and pharmaceutical compositions disclosed herein for use in the treatment of disorders or diseases mediated by the mTOR pathway, wherein the compounds require approximately 10-fold higher concentrations in FKBP12 KO cells compared to cells expressing FKBP12, and compared to the rapamycin analog RAD001, to achieve 20% inhibition of S6K1 cell signaling.

[0100] In some other embodiments, the compounds and pharmaceutical compositions disclosed herein for use in the treatment of disorders or diseases mediated by the mTOR pathway, wherein the compounds require approximately 100-fold higher concentrations in FKBP12 KO cells compared to cells expressing FKBP12, and compared to the rapamycin analog RAD001, to achieve 20% inhibition of S6K1 cell signaling.

[0101] In some other embodiments, the compounds and pharmaceutical compositions disclosed herein for use in the treatment of disorders or diseases mediated by the mTOR pathway, wherein the compounds require approximately 500-fold higher concentrations in FKBP12 KO cells compared to cells expressing FKBP12, and compared to the rapamycin analog RAD001, to achieve 20% inhibition of S6K1 cell signaling.

[0102] In some other embodiments, the compounds and pharmaceutical compositions disclosed herein for use in the treatment of disorders or diseases mediated by the mTOR pathway, wherein the compounds require approximately 1000 times higher concentrations in FKBP12 KO cells compared to cells expressing FKBP12, and compared to the rapamycin analog RAD001, to achieve 20% inhibition of S6K1 cell signaling.

[0103] In some other embodiments, the compounds and pharmaceutical compositions disclosed herein for use in the treatment of disorders or diseases mediated by the mTOR pathway, wherein the compounds require approximately 10-fold higher concentrations in FKBP12 KO cells compared to cells expressing FKBP12, and compared to the rapamycin analog RAD001, to achieve 30% inhibition of S6K1 cell signaling.

[0104] In some other embodiments, the compounds and pharmaceutical compositions disclosed herein for use in the treatment of disorders or diseases mediated by the mTOR pathway, wherein the compounds require approximately 100-fold higher concentrations in FKBP12 KO cells compared to cells expressing FKBP12, and compared to the rapamycin analog RAD001, to achieve 30% inhibition of S6K1 cell signaling.

[0105] In some other embodiments, the compounds and pharmaceutical compositions disclosed herein for use in the treatment of disorders or diseases mediated by the mTOR pathway, wherein the compounds require approximately 500-fold higher concentrations in FKBP12 KO cells compared to cells expressing FKBP12, and compared to the rapamycin analog RAD001, to achieve 30% inhibition of S6K1 cell signaling.

[0106] In some other embodiments, the compounds and pharmaceutical compositions disclosed herein for use in the treatment of disorders or diseases mediated by the mTOR pathway, wherein the compounds require approximately 1000 times higher concentrations in FKBP12 KO cells compared to cells expressing FKBP12, and compared to the rapamycin analog RAD001, to achieve 30% inhibition of S6K1 cell signaling.

[0107] In some other embodiments, the “FKBP12 selective rapamycin analog” is a rapamycin analog that is about 10 to about 1000 times less potent than RAD001 in cell lines that do not express FKBP12 (e.g., FKBP12 knockout cells compared to cells expressing FKBP12). As used herein, “potency” can be expressed as the concentration of rapamycin analog required to achieve 20% inhibition of S6K1(Thr389) phosphorylation in a cell-based assay, such as the assays used in Example 3 disclosed herein and Figures 1-11. For example, if the IC20 of a rapamycin analog in an assay for phosphorylation inhibition of S6K1 (Thr389) using an FKBP12 knockout cell line is at least 20x greater than the IC20 of RAD001 in the same assay using the same cell line expressing normal FKBP12 (e.g., 30x, 50x, 100x, 200x, 500x, 1000x greater), then the rapamycin analog is considered to be FKBP12 selective.

[0108] In some other embodiments, the “FKBP12 selective rapamycin analog” is a rapamycin analog that is about 10 to about 1000 times less potent than RAD001 in cell lines that do not express FKBP12 (e.g., FKBP12 knockout cells compared to cells expressing FKBP12). As used herein, “potency” can be expressed as the concentration of rapamycin analog required to achieve 30% inhibition of S6K1(Thr389) phosphorylation in a cell-based assay, such as the assays used in Example 3 disclosed herein and Figures 1-11. For example, if the IC30 of a rapamycin analog in an assay for phosphorylation inhibition of S6K1 (Thr389) using an FKBP12 knockout cell line is at least 20x greater than the IC30 of RAD001 in the same assay using the same cell line expressing normal FKBP12 (e.g., 30x, 50x, 100x, 200x, 500x, 1000x greater), then the rapamycin analog is considered to be FKBP12 selective.

[0109] In the implementation scheme, for example, the compound of formula (I) or a pharmaceutically acceptable salt thereof has a higher affinity for FKBP12 than rapamycin or RAD001, which is sufficient to inhibit mTORC1 binding.

[0110] In the implementation scheme, compared with rapamycin or RAD001, compounds of (I), (P-1), (P-2), (Ia)-(Ic) or (P-Ia)-(P-Ic) or their pharmaceutically acceptable salts can be complexed with FKBP12 to more effectively bind to and inhibit mTORC1.

[0111] In the implementation scheme, the higher affinity binding to FKBP12 results in higher efficacy, for example, compared to rapamycin or RAD001.

[0112] In the implementation plan, the efficacy of the treatment is determined based on experience, for example, compared with rapamycin or RAD001.

[0113] In another aspect, this disclosure provides a method for treating a subject with or previously determined to have FKBP12 levels sufficient to inhibit mTORc1, the method comprising administering to the subject in need a therapeutically effective amount of a (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic) compound, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition or combination of pharmaceuticals described herein.

[0114] In another aspect, this disclosure provides a method for treating an age-related disease or disorder in a subject with a corresponding need, the method comprising administering to the subject a therapeutically effective amount of a (I), (P-1), (P-2), (Ia)-(Ic) or (P-Ia)-(P-Ic) compound, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition or combination of pharmaceuticals described herein.

[0115] In the implementation plan, the disease or disorder is selected from sarcopenia, skin atrophy, cherry angioma, seborrheic keratosis, brain atrophy (also known as dementia), atherosclerosis, arteriosclerosis, emphysema, osteoporosis, osteoarthritis, hypertension, erectile dysfunction, cataracts, macular degeneration, glaucoma, stroke, cerebrovascular disease (stroke), chronic kidney disease, diabetic nephropathy, impaired liver function, liver fibrosis, autoimmune hepatitis, endometrial hyperplasia, metabolic dysfunction, renal vascular disease, hearing loss, and activity deficits (e.g., frailty). Cognitive decline, tendon stiffness, cardiac dysfunction such as cardiomegaly and / or systolic and / or diastolic dysfunction and / or hypertension, cardiac dysfunction leading to decreased ejection fraction, immunosenescence, Parkinson's disease, Alzheimer's disease, cancer, immunosenescence leading to cancer due to decreased immune surveillance, infections due to decreased immune function, chronic obstructive pulmonary disease (COPD), obesity, loss of taste, loss of smell, arthritis, and type II diabetes (including complications arising from diabetes such as kidney failure, blindness, and neurological disorders).

[0116] In another aspect, this disclosure provides a method for treating a disease or disorder in a subject with a corresponding need, the method comprising administering to the subject a therapeutically effective amount of a (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic) compound, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition, or a combination of pharmaceuticals thereof, wherein the disorder or disease is selected from:

[0117] • Acute or chronic organ or tissue graft rejection;

[0118] • Transplant vascular disease;

[0119] • Smooth muscle cell proliferation and migration lead to thickening of the vascular intima, vascular occlusion, obstructive coronary atherosclerosis, and restenosis;

[0120] • Autoimmune diseases and inflammatory conditions;

[0121] ·asthma;

[0122] Multidrug resistance (MDR);

[0123] Fungal infection;

[0124] Inflammation;

[0125] ·Infect;

[0126] Age-related diseases;

[0127] Neurodegenerative diseases;

[0128] • Proliferative disorders, such as cancer;

[0129] • Seizures and seizure-related disorders; and

[0130] • Mitochondrial myopathy and mitochondrial stress.

[0131] In another aspect, this disclosure provides a method for treating cancer in a subject with a corresponding need, the method comprising administering to the subject a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition or combination of pharmaceuticals described herein.

[0132] In the implementation plan, the method also includes a PD-1 / PDL-1 inhibitor.

[0133] In the implementation plan, the cancers are selected from kidney cancer, renal cell carcinoma, colorectal cancer, uterine sarcoma, endometrial cancer, breast cancer, ovarian cancer, cervical cancer, gastric cancer, fibrosarcoma, pancreatic cancer, liver cancer, melanoma, leukemia, multiple myeloma, nasopharyngeal carcinoma, prostate cancer, lung cancer, glioblastoma, bladder cancer, mesothelioma, head cancer, rhabdomyosarcoma, sarcoma, lymphoma, and cervical cancer.

[0134] In the implementation plan, the disorder is a liver disorder, which includes processes of fibrosis and / or inflammation, such as liver fibrosis occurring in end-stage liver disease; cirrhosis; liver failure due to toxicity; non-alcoholic steatosis or NASH; and alcoholic steatosis.

[0135] In the implementation plan, the disorder is a kidney disorder, which includes processes of fibrosis or inflammation in the kidneys, such as kidney fibrosis that occurs due to acute kidney injury, leading to chronic kidney disease and diabetic nephropathy.

[0136] In the implementation scheme, the disorder is a cardiac dysfunction, such as myocardial infarction or cardiac hypertrophy. In the implementation scheme, the cardiac dysfunction is systolic dysfunction and / or diastolic dysfunction. In the implementation scheme, the cardiac dysfunction is hypertension. In the implementation scheme, the cardiac dysfunction results in a decreased ejection fraction.

[0137] In the implementation plan, the disorder is due to immune senescence in cancer caused by a decline in immune surveillance.

[0138] In the implementation scheme, the disorder is cancer, including tumors treated with immunotherapy and tumors previously treated with rapamycin, RAD001, or another rapamycin analogue. In the implementation scheme, cancer includes tumors exhibiting mTOR pathway activation, including environments with mutations in the Tsc1 gene, or environments where the tumor microenvironment has been appropriately treated with a rapamycin analogue.

[0139] This document sets forth details of one or more embodiments of the present disclosure. Other features, objects, and advantages of the present disclosure will become apparent from the accompanying drawings, detailed description, embodiments, and claims. Brief description of the attached diagram

[0141] Figure 1A This is a line graph showing the inhibition of S6K1 (Thr389) in wild-type 293T cells. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles) and compound 44 (solid line with squares). All treatments were performed in duplicate. The Y-axis represents the percentage inhibition of S6K1 (Thr389) levels relative to cells treated with culture medium supplemented with dimethyl sulfoxide (DMSO). The X-axis represents the concentrations of the two compounds.

[0142] Figure 1B This is a line graph showing the inhibition of S6K1 (Thr389) in FKBP12 knockout 293T cells. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles) and compound 44 (solid line with squares). All treatments were performed in duplicate. The Y-axis represents the percentage inhibition of S6K1 (Thr389) levels relative to cells treated with culture medium supplemented with dimethyl sulfoxide (DMSO). The X-axis represents the concentrations of the two compounds.

[0143] Figure 1C This is a line graph showing the inhibition of S6K1 (Thr389) in wild-type 293T cells. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles) and compound 45 (solid line with squares). All treatments were performed in duplicate. The Y-axis represents the percentage inhibition of S6K1 (Thr389) levels relative to cells treated with culture medium supplemented with dimethyl sulfoxide (DMSO). The X-axis represents the concentrations of the two compounds.

[0144] Figure 1DThis is a line graph showing the inhibition of S6K1 (Thr389) in FKBP12 knockout 293T cells. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles) and compound 45 (solid line with squares). All treatments were performed in duplicate. The Y-axis represents the percentage inhibition of S6K1 (Thr389) levels relative to cells treated with culture medium supplemented with dimethyl sulfoxide (DMSO). The X-axis represents the concentrations of the two compounds.

[0145] Figure 2A This is a line graph showing the inhibition of S6K1 (Thr389) in wild-type 293T cells. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles) and compound 18 (solid line with squares). All treatments were performed in duplicate. The Y-axis represents the percentage inhibition of S6K1 (Thr389) levels relative to cells treated with culture medium supplemented with dimethyl sulfoxide (DMSO). The X-axis represents the concentrations of the two compounds.

[0146] Figure 2B This is a line graph showing the inhibition of S6K1 (Thr389) in FKBP12 knockout 293T cells. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles) and compound 18 (solid line with squares). All treatments were performed in duplicate. The Y-axis represents the percentage inhibition of S6K1 (Thr389) levels relative to cells treated with culture medium supplemented with dimethyl sulfoxide (DMSO). The X-axis represents the concentrations of the two compounds.

[0147] Figure 3A This is a line graph showing the inhibition of S6K1 (Thr389) in wild-type 293T cells. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles) and compound 16 (solid line with squares). All treatments were performed in duplicate. The Y-axis represents the percentage inhibition of S6K1 (Thr389) levels relative to cells treated with culture medium supplemented with dimethyl sulfoxide (DMSO). The X-axis represents the concentrations of the two compounds.

[0148] Figure 3B This is a line graph showing the inhibition of S6K1 (Thr389) in FKBP12 knockout 293T cells. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles) and compound 16 (solid line with squares). All treatments were performed in duplicate. The Y-axis represents the percentage inhibition of S6K1 (Thr389) levels relative to cells treated with culture medium supplemented with dimethyl sulfoxide (DMSO). The X-axis represents the concentrations of the two compounds.

[0149] Figure 4A This is a line graph showing the inhibition of S6K1 (Thr389) in wild-type 293T cells. Cells were treated for 2 hours with the following compounds: RAD001 (everolimus; dashed line with circles), compound 26 (solid line with squares), and compound 28 (solid line with crosses). All treatments were performed in duplicate. The Y-axis represents the percentage inhibition of S6K1 (Thr389) levels relative to cells treated with culture medium supplemented with dimethyl sulfoxide (DMSO). The X-axis represents the concentrations of the two compounds.

[0150] Figure 4B This is a line graph showing the inhibition of S6K1 (Thr389) in FKBP12 knockout 293T cells. Cells were treated for 2 hours with the following compounds: RAD001 (everolimus; dashed line with circles), compound 26 (solid line with squares), and compound 28 (solid line with crosses). All treatments were performed in duplicate. The Y-axis represents the percentage inhibition of S6K1 (Thr389) levels relative to cells treated with culture medium supplemented with dimethyl sulfoxide (DMSO). The X-axis represents the concentrations of the two compounds.

[0151] Figure 5A This is a line graph showing the inhibition of S6K1 (Thr389) phosphorylation in wild-type (WT) 293T cells with increasing compound treatment concentrations. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles); and compound 47 (solid line with squares). All treatments were performed in duplicate. The Y-axis shows the S6K1 (Thr389) phosphorylation value, calculated as a percentage (%) of the value in control cells treated only with medium supplemented with dimethyl sulfoxide (DMSO). The S6K1 (Thr389) phosphorylation value in control DMSO-treated cells was set at 100%. The X-axis represents the Log10 molar (M) concentration of the compound.

[0152] Figure 5B This is a line graph showing the inhibition of S6K1 (Thr389) phosphorylation in FKBP12 knockout (KO)293T cells with increasing compound treatment concentration. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles); and compound 47 (solid line with squares). All treatments were performed in duplicate. The Y-axis shows the S6K1 (Thr389) phosphorylation value, calculated as a percentage (%) of the value in control cells treated only with medium supplemented with dimethyl sulfoxide (DMSO). The S6K1 (Thr389) phosphorylation value in control DMSO-treated cells was set at 100%. The X-axis represents the Log10 molar (M) concentration of the compound.

[0153] Figure 5CThis is a line graph showing the inhibition of S6K1 (Thr389) phosphorylation in wild-type (WT) 293T cells with increasing compound treatment concentrations. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles); and compound 48 (solid line with squares). All treatments were performed in duplicate. The Y-axis shows the S6K1 (Thr389) phosphorylation value, calculated as a percentage (%) of the value in control cells treated only with medium supplemented with dimethyl sulfoxide (DMSO). The S6K1 (Thr389) phosphorylation value in control DMSO-treated cells was set at 100%. The X-axis represents the Log10 molar (M) concentration of the compound.

[0154] Figure 5D This is a line graph showing the inhibition of S6K1 (Thr389) phosphorylation in FKBP12 knockout (KO)293T cells with increasing compound treatment concentration. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles); and compound 48 (solid line with squares). All treatments were performed in duplicate. The Y-axis shows the S6K1 (Thr389) phosphorylation value, calculated as a percentage (%) of the value in control cells treated only with medium supplemented with dimethyl sulfoxide (DMSO). The S6K1 (Thr389) phosphorylation value in control DMSO-treated cells was set at 100%. The X-axis represents the Log10 molar (M) concentration of the compound.

[0155] Figure 5E This is a line graph showing the inhibition of S6K1 (Thr389) phosphorylation in wild-type (WT) 293T cells with increasing compound treatment concentrations. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles); and compound 49 (solid line with squares). All treatments were performed in duplicate. The Y-axis shows the S6K1 (Thr389) phosphorylation value, calculated as a percentage (%) of the value in control cells treated only with medium supplemented with dimethyl sulfoxide (DMSO). The S6K1 (Thr389) phosphorylation value in control DMSO-treated cells was set at 100%. The X-axis represents the concentrations of the two compounds.

[0156] Figure 5FThis is a line graph showing the inhibition of S6K1 (Thr389) phosphorylation in FKBP12 knockout (KO)293T cells with increasing compound treatment concentration. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles); and compound 49 (solid line with squares). All treatments were performed in duplicate. The Y-axis shows the S6K1 (Thr389) phosphorylation value, calculated as a percentage (%) of the value in control cells treated only with medium supplemented with dimethyl sulfoxide (DMSO). The S6K1 (Thr389) phosphorylation value in control DMSO-treated cells was set at 100%. The X-axis represents the Log10 molar (M) concentration of the two compounds.

[0157] Figure 6A This is a line graph showing the inhibition of S6K1 (Thr389) phosphorylation in wild-type (WT) 293T cells with increasing compound treatment concentrations. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles); and compound 50 (solid line with squares). All treatments were performed in duplicate. The Y-axis shows the S6K1 (Thr389) phosphorylation value, calculated as a percentage (%) of the value in control cells treated only with medium supplemented with dimethyl sulfoxide (DMSO). The S6K1 (Thr389) phosphorylation value in control DMSO-treated cells was set at 100%. The X-axis represents the concentrations of the two compounds.

[0158] Figure 6B This is a line graph showing the inhibition of S6K1 (Thr389) phosphorylation in FKBP12 knockout (KO)293T cells with increasing compound treatment concentration. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles); and compound 50 (solid line with squares). All treatments were performed in duplicate. The Y-axis shows the S6K1 (Thr389) phosphorylation value, calculated as a percentage (%) of the value in control cells treated only with medium supplemented with dimethyl sulfoxide (DMSO). The S6K1 (Thr389) phosphorylation value in control DMSO-treated cells was set at 100%. The X-axis represents the concentration of the two compounds.

[0159] Figure 6CThis is a line graph showing the inhibition of S6K1 (Thr389) phosphorylation in wild-type (WT) 293T cells with increasing compound treatment concentrations. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles); and compound 51 (solid line with squares). All treatments were performed in duplicate. The Y-axis shows the S6K1 (Thr389) phosphorylation value, calculated as a percentage (%) of the value in control cells treated only with medium supplemented with dimethyl sulfoxide (DMSO). The S6K1 (Thr389) phosphorylation value in control DMSO-treated cells was set at 100%. The X-axis represents the concentrations of the two compounds.

[0160] Figure 6D This is a line graph showing the inhibition of S6K1 (Thr389) phosphorylation in FKBP12 knockout (KO)293T cells with increasing compound treatment concentration. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles); and compound 51 (solid line with squares). All treatments were performed in duplicate. The Y-axis shows the S6K1 (Thr389) phosphorylation value, calculated as a percentage (%) of the value in control cells treated only with medium supplemented with dimethyl sulfoxide (DMSO). The S6K1 (Thr389) phosphorylation value in control DMSO-treated cells was set at 100%. The X-axis represents the concentration of the two compounds.

[0161] Figure 6E This is a line graph showing the inhibition of S6K1 (Thr389) phosphorylation in wild-type (WT) 293T cells with increasing compound treatment concentrations. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles); and compound 52 (solid line with squares). All treatments were performed in duplicate. The Y-axis shows the S6K1 (Thr389) phosphorylation value, calculated as a percentage (%) of the value in control cells treated only with medium supplemented with dimethyl sulfoxide (DMSO). The S6K1 (Thr389) phosphorylation value in control DMSO-treated cells was set at 100%. The X-axis represents the concentrations of the two compounds.

[0162] Figure 6FThis is a line graph showing the inhibition of S6K1 (Thr389) phosphorylation in FKBP12 knockout (KO)293T cells with increasing compound treatment concentrations. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles); and compound 52 (solid line with squares). All treatments were performed in duplicate. The Y-axis shows the S6K1 (Thr389) phosphorylation value, calculated as a percentage (%) of the value in control cells treated only with medium supplemented with dimethyl sulfoxide (DMSO). The S6K1 (Thr389) phosphorylation value in control DMSO-treated cells was set at 100%. The X-axis represents the concentrations of the two compounds.

[0163] Figure 7A This is a line graph showing the inhibition of S6K1 (Thr389) phosphorylation in wild-type (WT) 293T cells with increasing compound treatment concentrations. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles); and compound 57 (solid line with squares). All treatments were performed in duplicate. The Y-axis shows the S6K1 (Thr389) phosphorylation value, calculated as a percentage (%) of the value in control cells treated only with medium supplemented with dimethyl sulfoxide (DMSO). The S6K1 (Thr389) phosphorylation value in control DMSO-treated cells was set at 100%. The X-axis represents the concentrations of the two compounds.

[0164] Figure 7B This is a line graph showing the inhibition of S6K1 (Thr389) phosphorylation in FKBP12 knockout (KO)293T cells with increasing compound treatment concentrations. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles); and compound 57 (solid line with squares). All treatments were performed in duplicate. The Y-axis shows the S6K1 (Thr389) phosphorylation value, calculated as a percentage (%) of the value in control cells treated only with medium supplemented with dimethyl sulfoxide (DMSO). The S6K1 (Thr389) phosphorylation value in control DMSO-treated cells was set at 100%. The X-axis represents the concentrations of the two compounds.

[0165] Figure 7CThis is a line graph showing the inhibition of S6K1 (Thr389) phosphorylation in wild-type (WT)293T cells with increasing compound treatment concentrations. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles); and compound 58 (solid line with squares). All treatments were performed in duplicate. The Y-axis shows the S6K1 (Thr389) phosphorylation value, calculated as a percentage (%) of the value in control cells treated only with medium supplemented with dimethyl sulfoxide (DMSO). The S6K1 (Thr389) phosphorylation value in control DMSO-treated cells was set at 100%. The X-axis represents the concentrations of the two compounds.

[0166] Figure 7D This is a line graph showing the inhibition of S6K1 (Thr389) phosphorylation in FKBP12 knockout (KO)293T cells with increasing compound treatment concentration. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles); and compound 58 (solid line with squares). All treatments were performed in duplicate. The Y-axis shows the S6K1 (Thr389) phosphorylation value, calculated as a percentage (%) of the value in control cells treated only with medium supplemented with dimethyl sulfoxide (DMSO). The S6K1 (Thr389) phosphorylation value in control DMSO-treated cells was set at 100%. The X-axis represents the concentration of the two compounds.

[0167] Figure 7E This is a line graph showing the inhibition of S6K1 (Thr389) phosphorylation in wild-type (WT)293T cells with increasing compound treatment concentrations. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles); and compound 59 (solid line with squares). All treatments were performed in duplicate. The Y-axis shows the S6K1 (Thr389) phosphorylation value, calculated as a percentage (%) of the value in control cells treated only with medium supplemented with dimethyl sulfoxide (DMSO). The S6K1 (Thr389) phosphorylation value in control DMSO-treated cells was set at 100%. The X-axis represents the concentrations of the two compounds.

[0168] Figure 7FThis is a line graph showing the inhibition of S6K1 (Thr389) phosphorylation in FKBP12 knockout (KO)293T cells with increasing compound treatment concentrations. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles); and compound 59 (solid line with squares). All treatments were performed in duplicate. The Y-axis shows the S6K1 (Thr389) phosphorylation value, calculated as a percentage (%) of the value in control cells treated only with medium supplemented with dimethyl sulfoxide (DMSO). The S6K1 (Thr389) phosphorylation value in control DMSO-treated cells was set at 100%. The X-axis represents the concentrations of the two compounds.

[0169] Figure 8A This is a line graph showing the inhibition of S6K1 (Thr389) phosphorylation in wild-type (WT) 293T cells with increasing compound treatment concentrations. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles); and compound 63 (solid line with squares). All treatments were performed in duplicate. The Y-axis shows the S6K1 (Thr389) phosphorylation value, calculated as a percentage (%) of the value in control cells treated only with medium supplemented with dimethyl sulfoxide (DMSO). The S6K1 (Thr389) phosphorylation value in control DMSO-treated cells was set at 100%. The X-axis represents the concentrations of the two compounds.

[0170] Figure 8B This is a line graph showing the inhibition of S6K1 (Thr389) phosphorylation in FKBP12 knockout (KO)293T cells with increasing compound treatment concentration. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles); and compound 63 (solid line with squares). All treatments were performed in duplicate. The Y-axis shows the S6K1 (Thr389) phosphorylation value, calculated as a percentage (%) of the value in control cells treated only with medium supplemented with dimethyl sulfoxide (DMSO). The S6K1 (Thr389) phosphorylation value in control DMSO-treated cells was set at 100%. The X-axis represents the concentration of the two compounds.

[0171] Figure 9AThis is a line graph showing the inhibition of S6K1 (Thr389) phosphorylation in wild-type (WT) 293T cells with increasing compound treatment concentrations. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles); and compound 69 (solid line with squares). All treatments were performed in duplicate. The Y-axis shows the S6K1 (Thr389) phosphorylation value, calculated as a percentage (%) of the value in control cells treated only with medium supplemented with dimethyl sulfoxide (DMSO). The S6K1 (Thr389) phosphorylation value in control DMSO-treated cells was set at 100%. The X-axis represents the concentrations of the two compounds.

[0172] Figure 9B This is a line graph showing the inhibition of S6K1 (Thr389) phosphorylation in FKBP12 knockout (KO)293T cells with increasing compound treatment concentration. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles); and compound 69 (solid line with squares). All treatments were performed in duplicate. The Y-axis shows the S6K1 (Thr389) phosphorylation value, calculated as a percentage (%) of the value in control cells treated only with medium supplemented with dimethyl sulfoxide (DMSO). The S6K1 (Thr389) phosphorylation value in control DMSO-treated cells was set at 100%. The X-axis represents the concentration of the two compounds.

[0173] Figure 10A This is a line graph showing the inhibition of S6K1 (Thr389) phosphorylation in wild-type (WT) 293T cells with increasing compound treatment concentrations. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles); and compound 72 (solid line with squares). All treatments were performed in duplicate. The Y-axis shows the S6K1 (Thr389) phosphorylation value, calculated as a percentage (%) of the value in control cells treated only with medium supplemented with dimethyl sulfoxide (DMSO). The S6K1 (Thr389) phosphorylation value in control DMSO-treated cells was set at 100%. The X-axis represents the concentrations of the two compounds.

[0174] Figure 10BThis is a line graph showing the inhibition of S6K1 (Thr389) phosphorylation in FKBP12 knockout (KO)293T cells with increasing compound treatment concentrations. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles); and compound 72 (solid line with squares). All treatments were performed in duplicate. The Y-axis shows the S6K1 (Thr389) phosphorylation value, calculated as a percentage (%) of the value in control cells treated only with medium supplemented with dimethyl sulfoxide (DMSO). The S6K1 (Thr389) phosphorylation value in control DMSO-treated cells was set at 100%. The X-axis represents the concentrations of the two compounds.

[0175] Figure 11A This is a line graph showing the inhibition of S6K1 (Thr389) phosphorylation in wild-type (WT) 293T cells with increasing compound treatment concentrations. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles); and compound 73 (solid line with squares). All treatments were performed in duplicate. The Y-axis shows the S6K1 (Thr389) phosphorylation value, calculated as a percentage (%) of the value in control cells treated only with medium supplemented with dimethyl sulfoxide (DMSO). The S6K1 (Thr389) phosphorylation value in control DMSO-treated cells was set at 100%. The X-axis represents the concentrations of the two compounds.

[0176] Figure 11B This is a line graph showing the inhibition of S6K1 (Thr389) phosphorylation in FKBP12 knockout (KO)293T cells with increasing compound treatment concentration. Cells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed line with circles); and compound 73 (solid line with squares). All treatments were performed in duplicate. The Y-axis shows the S6K1 (Thr389) phosphorylation value, calculated as a percentage (%) of the value in control cells treated only with medium supplemented with dimethyl sulfoxide (DMSO). The S6K1 (Thr389) phosphorylation value in control DMSO-treated cells was set at 100%. The X-axis represents the concentration of the two compounds.

[0177] Figure 12A This is a graph showing the plasma pharmacokinetic profile of compound RAD001 in mice after IV (2 mg / kg) and PO (10 mg / kg) administration. The Y-axis, set to a log10 scale, represents the concentration of the compound in plasma (ng / ml). The X-axis represents the time points (hours) of plasma collection after compound administration.

[0178] Figure 12BThis is a graph showing the plasma pharmacokinetic profile of compound 26 in mice after IV (2 mg / kg) and PO (20 mg / kg) administration. The Y-axis, set to a log10 scale, represents the plasma concentration of the compound (ng / ml). The X-axis represents the time points (hours) of plasma collection after compound administration.

[0179] Figure 12C This is a graph showing the pharmacokinetic profile of compound 45 in mouse plasma after IV (2 mg / kg) and PO (20 mg / kg) administration. The Y-axis, set to a log10 scale, represents the concentration of the compound in plasma, ng / ml. The X-axis represents the time points (hours) of plasma collection after compound administration.

[0180] Figure 12D This is a graph showing the pharmacokinetic profile of compound 69 in mouse plasma after IV (2 mg / kg) and PO (10 mg / kg) administration. The Y-axis, set to a log10 scale, represents the concentration of the compound in plasma, ng / ml. The X-axis represents the time points (hours) of plasma collection after compound administration.

[0181] Pharmacokinetic studies of the selected compounds in mice Figures 12A-12D The data described in the figure are further summarized in Table 4 shown in the following detailed description. Invention Details

[0183] Certainly

[0184] When referring to the compounds provided herein, the following terms have the following meanings unless otherwise indicated. Unless otherwise defined, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art. Where multiple definitions exist for terms provided herein, those definitions shall prevail unless otherwise stated.

[0185] As used herein, “alkyl” refers to a monovalent and saturated hydrocarbon group moiety. Alkyl groups are optionally substituted and can be straight-chain, branched, or cyclic (i.e., cycloalkyl). Alkyl groups include, but are not limited to, those groups having 1 to 20 carbon atoms, such as C… 1-20 Alkyl groups; those groups having 1 to 12 carbon atoms, for example, C 1-12 Alkyl groups; those groups having 1 to 8 carbon atoms, for example, C 1-8 Alkyl groups; those groups having 1 to 6 carbon atoms, for example, C 1-6 Alkyl groups; and those groups having 1 to 3 carbon atoms, for example, C 1-3Alkyl groups. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, isobutyl, pentyl, hexyl, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. The pentyl group includes, but is not limited to, n-pentyl and isopentyl. The hexyl group includes, but is not limited to, n-hexyl.

[0186] As used herein, “halogenated alkyl” means an alkyl group as defined herein, wherein the alkyl group includes at least one substituent selected from halogens (e.g., F, Cl, Br or I).

[0187] As used herein, “hydroxyalkyl” means an alkyl group as defined herein, wherein the alkyl group comprises at least one hydroxyl group.

[0188] As used herein, "alkylene" refers to a divalent alkyl group. Unless otherwise stated, alkylene groups include, but are not limited to, those with 1 to 20 carbon atoms. Alkylene groups are optionally substituted, as described herein with respect to alkyl groups. In some embodiments, the alkylene group is unsubstituted. Examples of alkylene moiety include -CH2-, -CH2CH2-, -CH2CH2CH2-, -CH2CH2CH2CH2-, etc.

[0189] Specifying an amino acid or amino acid residue without specifying a stereochemical intent covers the L-form, D-form, or racemic mixture of an amino acid or amino acid residue.

[0190] As used herein, “alkoxy” refers to a monovalent and saturated hydrocarbon group moiety, wherein the hydrocarbon comprises a single bond with an oxygen atom, and wherein the group is located on the oxygen atom, for example, CH3CH2-O- represents ethoxy. Alkoxy substituents are bonded to the compounds they substituted through this oxygen atom of the alkoxy substituent. Alkoxy groups are optionally substituted and can be straight-chain, branched, or cyclic, for example, cycloalkoxy groups. Alkoxy groups include, but are not limited to, those having 1 to 20 carbon atoms, for example, C 1-20 Alkoxy groups; those groups having 1 to 12 carbon atoms, for example, C 1-12 Alkoxy groups; those groups having 1 to 8 carbon atoms, for example, C 1-8 Alkoxy groups; those groups having 1 to 6 carbon atoms, for example, C 1-6 Alkoxy groups; and those groups having 1 to 3 carbon atoms, for example, C 1-3 Alkoxy groups. Examples of alkoxy moieties include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, isobutoxy, pentoxy moieties, hexoxy moieties, cyclopropoxy, cyclobutoxy, cyclopentoxy, and cyclohexoxy.

[0191] As used herein, “haloalkoxy” means an alkoxy group as defined herein, wherein the hydrocarbon is substituted with at least one halogen (e.g., F, Cl, Br or I).

[0192] As used in this article, "aryl" refers to a compound with a unit price of C5-C. 15 A carbocyclic system comprising at least one aromatic ring, wherein the aryl ring system is monocyclic, bicyclic, or tricyclic. The aryl group can be attached to the main structure via any of its rings (i.e., any aromatic or non-aromatic ring). In some or any embodiment, the aryl group can be a bridged (where chemically feasible) or non-bridged, spirocyclic (where chemically feasible) or non-spirocyclic, and / or fused or non-fused polycyclic group. In some or any embodiment, the aryl group is phenyl, naphthyl, bicyclic [4.2.0]octyl-1,3,5-trienyl, indanyl, fluorenyl, 6,7,8,9-tetrahydro-5H-benzo[7]arunyl, Or tetrahydronaphthyl. When the aryl group is substituted, it can be substituted on any ring, that is, on any aromatic or non-aromatic ring contained in the aryl group. In some or any embodiment, the aryl group is phenyl, naphthyl, tetrahydronaphthyl, fluorenyl, 6,7,8,9-tetrahydro-5H-benzo[7]arunenyl or indanyl.

[0193] As used herein, "heteroaryl" refers to a monocyclic aromatic ring system or a polycyclic aromatic ring system, wherein one or more ring atoms (in some or any embodiment, 1, 2, 3 or 4) are independently selected from O, S(O). 0-2 The heteroaryl group has heteroatoms of NH and N, and the remaining ring atoms are carbon atoms, wherein the rings can optionally be substituted as described herein. Where permitted by the valence rules, the heteroaryl group is bonded to the remainder of the molecule via any atom in the ring system. In some embodiments, each ring of the heteroaryl group may contain one or two O atoms, one or two S atoms, and / or one to four N atoms, or combinations thereof, provided that the total number of heteroatoms in each ring is four or fewer and each ring contains at least one carbon atom. In some embodiments, the heteroaryl group has from 5 to 20, from 5 to 15, or from 5 to 10 ring atoms. When the heteroaryl group is substituted, it can be substituted on any ring. In some embodiments, the heteroaryl group includes, but is not limited to, furanyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, thiophenyl, pyridyl, pyrimidinyl, pyridazinyl, indoleyl, quinolinyl, isoquinolinyl, oxazolyl, isoxazolyl, thienopyridyl, thienopyrimidinyl, and azaindolineyl. In some embodiments, the heteroaryl group is... in This indicates the attachment point of the heteroaryl group to the rest of the molecule. In some embodiments, when substituted with one or more hydroxyl groups, the heteroaryl group can be named or drawn as a ketone or enol tautomer. For example, 2,4(1H,3H)-dioxo-pyrimidinyl, 2,4-dihydroxy-pyrimidinyl, 2(1H)-oxo-4-hydroxypyrimidinyl, 4-hydroxy-2(3H)-oxo-pyrimidinyl, and 2-hydroxy-4(3H)-oxo-pyrimidinyl are within the range of heteroaryl groups when substituted with two hydroxyl groups. In some embodiments, "heteroaryl" is N-linked.

[0194] In some embodiments, the monocyclic heteroaryl group includes, but is not limited to, furanyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridinyl, pyrimidinyl, pyrroleyl, thiadiazolyl, thiazolyl, thiophene, tetrazolyl, triazinyl, and triazolyl. In some embodiments, the bicyclic heteroaryl group includes, but is not limited to, benzofuranyl, benzimidazolyl, benzoisoxazolyl, benzopyranyl, benzothiadiazolyl, benzothiazolyl, benzothiophenyl, benzotriazolyl, benzooxazolyl, furanpyridyl, imidazopyridyl, imidazothiazolyl, indazinyl, indolyl, indazolyl, isobenzofuranyl, isobenzothiaphenyl, isoindolyl, isoquinolinyl, isothiazolyl, naphridinyl, oxazolopyridyl, phthalazinyl, pteridinyl, purine, pyridopyridyl, pyrrolopyridyl, quinolinyl, quinoxalinyl, quinazolinyl, thiadiazolopyrimidinyl, and thiophenopyridyl. In some embodiments, the tricyclic heteroaryl group includes, but is not limited to, acridinel, benzoindolyl, carbazolel, dibenzofuranyl, perimidinyl, phenanthrolinyl, phenanthridinyl, and phenazinyl. In some or any embodiments, the heteroaryl group is indolyl, furanyl, pyridyl, pyrimidinyl, imidazolyl, or pyrazolyl; each optionally substituted by one, two, three, or four groups as defined throughout the specification, including, in some embodiments, substitution by groups independently selected from: C 1-6 Alkyl, hydroxyl, halogen, halogen-C 1-6 Alkyl, C 1-6 Alkoxy, cyano, or phenyl.

[0195] As used herein, "heterocyclic alkyl" or "heterocyclic group" refers to a monovalent monocyclic nonaromatic ring system and / or a polycyclic system containing at least one nonaromatic ring; wherein one or more nonaromatic monocyclic ring atoms (in some or any embodiments, 1, 2, 3 or 4) are independently selected from O, S(O). 0-2The polycyclic system contains heteroatoms of N, and the remaining ring atoms are carbon atoms; and one or more of any ring atoms in the polycyclic system (in some or any embodiment, 1, 2, 3 or 4) are independently selected from O, S (O). 0-2 The heterocycle contains a nitrogen atom and the remaining ring atoms are carbon. The term "heterocycle" does not include fully aromatic rings, i.e., it does not include imidazoles, pyrimidines, pyridines, etc. In some or any embodiments, the heterocycle contains one or two heteroatoms independently selected from nitrogen and oxygen. In some or any embodiments, the heterocycle contains one or two oxygen heteroatoms. In some or any embodiments, the heterocycle contains one or two nitrogen heteroatoms (wherein nitrogen is substituted as described in any aspect or embodiment herein). In some or any embodiments, the heterocycle is polycyclic and contains one heteroatom in a non-aromatic ring, or contains one heteroatom in an aromatic ring, or contains two heteroatoms in an aromatic ring, or contains two heteroatoms, one in an aromatic ring and the other in a non-aromatic ring. In some or any embodiments, the heterocyclic group has 3 to 20, 3 to 15, 3 to 10, 3 to 8, 4 to 7, or 5 to 6 ring atoms. In some or any embodiment, the heterocycle is a monocyclic, bicyclic, tricyclic, or tetracyclic ring system. In some or any embodiment, the heterocyclic group can be a bridged or unbridged, spirocyclic or non-spirocyclic, and / or fused or non-fused polycyclic group. One or more of the nitrogen and sulfur atoms can optionally be oxidized, one or more of the nitrogen atoms can optionally be quaternized, and one or more of the carbon atoms can optionally be replaced with... Some rings may be partially or fully saturated, or aromatic, provided that the heterocycle is not fully aromatic. Monocyclic and polycyclic heterocycles can be attached to the host structure at any heteroatom or carbon atom, resulting in stable compounds. Polycyclic heterocycles can be attached to the host structure by any of their rings (including any aromatic or non-aromatic rings), regardless of whether that ring contains a heteroatom. In some or any embodiment, the heterocycle is a “heterocyclic alkyl group”, which is 1) a saturated or partially unsaturated (but not aromatic) monovalent monocyclic group containing at least one ring heteroatom, as described herein, or 2) a saturated or partially unsaturated (but not aromatic) monovalent bicyclic or tricyclic group, wherein at least one ring contains at least one heteroatom, as described herein. When heterocycles and heterocyclic alkyl groups are substituted, they can be substituted on any ring, i.e., on any aromatic or non-aromatic ring contained in the heterocycle and heterocyclic alkyl group. In some or any embodiment, such heterocycles include, but are not limited to, nitrogen-containing heterocycles. azepinyl, benzodioxane-hexyl, benzodioxane-pentenyl, 3,4-dihydro-2H-benzo[b][1,4]oxazinyl, 3,4-dihydro-2H-benzo[b][1,4]dioxane-heptenyl, 1,3-dihydroisobenzofuranyl, benzofuranone, benzopyranone, benzopyranyl, dihydrobenzofuranyl Benzotetrahydrothiophene, 2,2-dioxo-1,3-dihydrobenzo[c]thiophene, benzothiopyranyl, benzooxazinyl, β-carbolinyl, chromanyl, chromonelyl, cenolinyl, coumarinyl, decahydroquinolinyl, decahydroisoquinolinyl, dihydrobenzisothiazinyl, dihydrobenzisothiazinyl, dihydrofuranyl, dihydroisoindolyl, dihydropyranyl, dihydropyrazolyl, dihydropyridyl, dihydropyridyl, dihydropyrimidinyl, dihydropyrimidinyl Dihydropyrrole, dioxacyclopentyl, 1,4-dithianyl, furanone, imidazoalkyl, 2,4-dioxo-imidazoalkyl, imidazolinyl, indololinyl, 2-oxo-indololinyl, isobenzotetrahydrofuranyl, isobenzotetrahydrothiophenyl, isochoryl, isocoumarinyl, isoindololinyl, 1-oxo-isoindololinyl, 1,3-dioxo-isoindololinyl alkyl, isothiazolyl, isoxazolyl, 3-oxo-isooxazolyl, morpholinyl, 3,5-dioxo-morpholinyl, octahydroindolyl, octahydroisoindolyl, 1-oxo-octahydroisoindolyl, 1,3-dioxo-hexahydroisoindolyl, oxazolidinone, oxazolyl, ethylene oxide, piperazine, 2,6-dioxo-piperazine, piperidinyl, 2,6-dioxo-piperidinyl, 4-piperidinone, pyrazolyl Pyrazolinyl, pyrrolidinyl, pyrrolidinyl, 2-oxopyrrolidinyl, 2,5-dioxopyrrolidinyl, quininecycloyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydropyranyl, tetrahydrothiophenyl, thiomorpholinyl, 3,5-dioxo-thiomorpholinyl, thiazolyl, 2,4-dioxo-thiazolyl, tetrahydroquinolinyl, phenothiazinyl, phenotoxazinyl, xanthonyl, and 1,3,5-trithiaalkyl. In some or any embodiments, the heterocycle is benzo-1,4-dioxane-hexyl, benzodioxane-pentenyl, indololinyl, 2-oxo-indololinyl, pyrrolyl, piperidinyl, 2,3-dihydrobenzofuranyl, or decahydroquinolinyl; each optionally substituted with one, two, three, or four groups as defined throughout the specification, including substitution with groups independently selected from halogens, alkyl groups, and phenyl groups in some or any embodiments. In some embodiments, the heterocyclic alkyl group is pyrrolyl.

[0196] As used in this article, "cyano" refers to -CN.

[0197] Unless otherwise stated, the term "oxo" as used herein refers to a ketone group (C=O). An oxo group, as a substituent for a non-aromatic carbon, results in a conversion from -CH₂- to -C=O. An oxo group, as a substituent for an aromatic carbon, results in a conversion from -CH- to -C=O. When the substituent is oxo, two hydrogen atoms on the atom are replaced. When an oxo group replaces an aromatic moiety, the corresponding partially unsaturated ring replaces the aromatic ring. For example, a pyridyl group replaced by an oxo group is a pyridinium. Those skilled in the art will understand that in some embodiments, such groups, such as pyridinium and 2,4(1H,3H)-dioxo-pyrimidinyl, may exist in their tautomeric forms, such as hydroxypyridine and 2,4-dihydroxypyrimidinyl, respectively.

[0198] As used herein, when describing a group moiety, such as an optionally substituted alkyl group, "optionally substituted" means that such a moiety is optionally bonded to one or more substituents. Examples of such substituents include, but are not limited to, halogens, cyano groups, nitro groups, amino groups, hydroxyl groups, optionally substituted haloalkyl groups, aminoalkyl groups, hydroxyalkyl groups, azide groups, epoxy groups, optionally substituted heteroaryl groups, and optionally substituted heterocyclic alkyl groups. Where R A R B and R C Each time it appears independently, it is hydrogen, alkyl, alkenyl, alkynyl, aryl, alkylaryl, arylalkyl, heteroalkyl, heteroaryl, or heterocyclic alkyl, or R. A and R B Together with the atoms to which they are bonded, they form a saturated or unsaturated carbocyclic ring, wherein the ring is optionally substituted, and one or more ring atoms are optionally replaced by heteroatoms. In some embodiments, when the group is optionally substituted with a optionally substituted heteroaryl group, optionally substituted heterocycloalkyl group, or optionally substituted saturated or unsaturated carbocyclic ring, the substituents on the optionally substituted heteroaryl group, optionally substituted heterocycloalkyl group, or optionally substituted saturated or unsaturated carbocyclic ring (if they are substituted) are not substituted by substituents that are further optionally substituted by other substituents. In some embodiments, when the group described herein is optionally substituted, the substituents bonded to the group are unsubstituted unless otherwise stated.

[0199] As used herein, the term "diastereomer" is one of a pair of diastereomers that has opposite configurations only at one of at least two stereoisomer source centers or chiral centers. For example, compounds of formula (I), (Ia), (Ib), or (Ic) having (R)-stereochemistry at C-16 are diastereomers of the corresponding compounds having (S)-stereochemistry.

[0200] As used herein, “diasteremeric excess (de)” refers to a dimensionless molar ratio describing the purity of a chiral substance containing more than one stereoisomer source center. For example, zero diasteremeric excess would indicate an equimolar mixture of diastereomers. By another example, 99% diasteremeric excess would indicate a nearly stereopure diastereomer compound (i.e., a significant excess of one diastereomer relative to another). Diasteremeric excess can be calculated using a method similar to ee. As those skilled in the art will understand, de is typically reported as a percentage de (%de). %de can be calculated in a manner similar to %ee.

[0201] As used herein, "therapeutic effective amount" means an amount (e.g., the amount of a compound) sufficient to provide a therapeutic benefit to a patient or to delay or minimize one or more symptoms associated with a disease or disorder in the treatment or management of the disease or disorder.

[0202] Certain groups, parts, substituents, and atoms are depicted with wavy lines intersecting one or more bonds to indicate the group, part, substituent, or atom through the atoms to which it is bonded. For example, it is depicted as: The propyl group substituted with the phenyl group has the following structure: As used herein, unless otherwise stated, diagrams illustrating substituents bonded to cyclic groups (e.g., aromatic, heteroaromatic, fused-ring, and saturated or unsaturated cycloalkyl or heterocycloalkyl) via inter-atom bonds are schematic to indicate that, according to the techniques set forth herein or known in the art to which this disclosure pertains, the cyclic group may be substituted at any ring position in the cyclic group or on any ring in a fused-ring group. For example, groups Where the subscript q is an integer from 0 to 2, and where the substituent R 2a The position is generally described, for example, not directly attached to any vertex of the bond line structure, such as a specific cyclic carbon atom, including non-limiting examples of groups where the substituent R 2a Bonded to specific ring carbon atoms:

[0203] FKB12 ​​selective rapamycin analogue

[0204] As used in this article, "FKBP12 selectivity" refers to the requirement for FKBP12 to inhibit mTORC1 phosphorylation of S6K1. Figure 1CAs shown in the figure; compound 45 is able to inhibit S6K1 phosphorylation in the presence of FKBP12; however, when FKBP12 is knocked out, compound 45 has much less inhibition of S6K1 phosphorylation than RAD001, a rapamycin analog that can exert its effects through other FKBPs.

[0205] Alternatively, “FKBP12 selective rapamycin analog” is a rapamycin analog that, while effective in assays using wild-type cells, is approximately 10-fold to approximately 1000-fold less potent than RAD001 in cell lines that do not express FKBP12 (e.g., FKBP12 knockout cells). As used in this context, “potency” can be expressed as the concentration of rapamycin analog required to achieve 20% inhibition of S6K1(Thr389) phosphorylation in cell-based assays such as those used in Example 3 and Figures 1-11. For example, if a rapamycin analog inhibits approximately 20% of S6K1(Thr389) phosphorylation in an assay using an FKBP12 knockout cell line, and its potency is at least 20x lower (e.g., 20x, 30x, 50x, 100x, 200x, 500x, 1000x, etc.) compared to a similar level of inhibition achieved by RAD001 in the same assay using the same cell line expressing normal FKBP12, then the rapamycin analog is considered to be FKBP12 selective.

[0206] Alternatively, “FKBP12 selective rapamycin analog” is a rapamycin analog that, while effective in assays using wild-type cells, is approximately 10-fold to approximately 1000-fold less potent than RAD001 in cell lines that do not express FKBP12 (e.g., FKBP12 knockout cells). As used in this context, “potency” can be expressed as the concentration of rapamycin analog required to achieve 30% inhibition of S6K1(Thr389) phosphorylation in cell-based assays such as those used in Example 3 and Figures 1-11. For example, if a rapamycin analog inhibits approximately 30% of S6K1 (Thr389) phosphorylation in an assay using an FKBP12 knockout cell line, and its potency is at least 20x lower (e.g., 20x, 30x, 50x, 100x, 200x, 500x, 1000x, etc.) compared to a similar level of inhibition achieved by RAD001 in the same assay using the same cell line expressing normal FKBP12, then the rapamycin analog is considered to be FKBP12 selective.

[0207] In one embodiment, the compound of formula (I) is a compound of formula (PI):

[0208]

[0209] Or its pharmaceutically acceptable salt;

[0210] Where R 1 It is hydrogen, C 1-6 Alkyl, heterocyclic, aryl, heteroaryl, -C 0-6 Alkylene-SO2R 4 or -C 0-6 Alkylene-SO2R 5 The heterocyclic group, aryl group, and heteroaryl group are optionally surrounded by one or two R groups. 1a Group substitution;

[0211] R 2 It is a heterocyclic group, aryl group, heteroaryl group, -C 0-6 Alkylene-SO2R 4 or -C 0-6 Alkylene-SO2R 5 ; wherein the heterocyclic group, aryl group, and heteroaryl group are optionally surrounded by one or two R groups. 2a Group substitution;

[0212] Or R 1 and R 2 Together with the attached nitrogen, it forms an N-linked heteroaryl group;

[0213] R 3 Choose from the following groups: - OR a 3-6 membered heterocyclic groups and 3-6 membered heteroaryl groups;

[0214] R a Choose from the following groups: H, -P(O)(R b )2、-C(O)R c -C(O)OR c C 1-6 Alkyl and C 1-6 Hydroxyalkyl;

[0215] Each R b Choose independently from the following groups: H and C 1-6 alkyl;

[0216] R c Choose from the following groups: H, C 1-6 Alkyl and C 1-6 Hydroxyalkyl;

[0217] R 4 It is C 2-6 Alkyl, C 3-6 cycloalkyl, C 1-6 hydroxyalkyl, heterocyclic, aryl, or heteroaryl; wherein the heterocyclic, aryl, and heteroaryl groups are optionally surrounded by one or two R groups. 4aGroup substitution; and

[0218] R 5 It is a heterocyclic, aryl, or heteroaryl group; wherein the heterocyclic, aryl, or heteroaryl group is optionally surrounded by one or two R groups. 5a Group substitution;

[0219] One or two R 1a R 2a R 4a and R 5a Each of the groups, when present, is independently selected from C. 1-6 Alkyl, C 1-6 Hydroxyalkyl, hydroxyl, halogen, C 1-6 Halogenated alkyl groups and C 1-6 alkoxy; or any two R 1a R 2a R 4a and R 5a When groups exist on the same carbon atom, they form oxo groups together.

[0220] Where R 1 When it is hydrogen, R 2 Not -C 0-6 Alkylene-SO2R 4 .

[0221] In one embodiment, the compound of formula (Ia) is a compound of formula (P-Ia):

[0222]

[0223] Or its pharmaceutically acceptable salt;

[0224] Where R 1 Is it hydrogen or C? 1-6 alkyl;

[0225] R 2 It is optional to be controlled by one or two Rs 2a Heterocyclic, aryl, or heteroaryl groups substituted with functional groups;

[0226] R 3 Choose from the following groups: - OR a 3-6 membered heterocyclic groups and 3-6 membered heteroaryl groups;

[0227] R a Choose from the following groups: H, -P(O)(R b )2、-C(O)R c -C(O)OR c C 1-6 Alkyl and C 1-6 Hydroxyalkyl;

[0228] Each R b Choose independently from the following groups: H and C 1-6 alkyl;

[0229] R c Choose from the following groups: H, C 1-6 Alkyl and C 1-6 Hydroxyalkyl; and

[0230] One or two R 2a Each of the groups, when present, is independently selected from C. 1-6 Alkyl, C 1-6 Hydroxyalkyl, hydroxyl, halogen, C 1-6 Halogenated alkyl groups and C 1-6 alkoxy; or any two R 2a When groups exist on the same carbon atom, they form oxo groups together.

[0231] In one embodiment, the compound of formula (Ib) is a compound of formula (P-Ib):

[0232]

[0233] Or its pharmaceutically acceptable salt;

[0234] Where R 1 and R 2 Together with the attached nitrogen, it forms an optional structure with one or two R 1a N-linked heteroaryl groups substituted with functional groups;

[0235] R 3 Choose from the following groups: - OR a 3-6 membered heterocyclic groups and 3-6 membered heteroaryl groups;

[0236] R a Choose from the following groups: H, -P(O)(R b )2、-C(O)R c -C(O)OR c C 1-6 Alkyl and C 1-6 Hydroxyalkyl;

[0237] Each R b Choose independently from the following groups: H and C 1-6 alkyl;

[0238] R c Choose from the following groups: H, C 1-6 Alkyl and C 1-6 Hydroxyalkyl;

[0239] One or two R1a Each of the groups, when present, is independently selected from C. 1-6 Alkyl, C 1-6 Hydroxyalkyl, hydroxyl, halogen, C 1-6 Halogenated alkyl groups and C 1-6 alkoxy group; or two R groups 1a When groups exist on the same carbon atom, they form oxo groups together.

[0240] In one embodiment, the compound of formula (Ic) is a compound of formula (P-Ic):

[0241]

[0242] Or its pharmaceutically acceptable salt;

[0243] Where R 1 Is it hydrogen or C? 1-6 alkyl;

[0244] R 2 It is -C 0-6 Alkylene-SO2R 4 or -C 0-6 Alkylene-SO2R 5 ;

[0245] R 3 Choose from the following groups: - OR a 3-6 membered heterocyclic groups and 3-6 membered heteroaryl groups;

[0246] R a Choose from the following groups: H, -P(O)(R b )2、-C(O)R c -C(O)OR c C 1-6 Alkyl and C 1-6 Hydroxyalkyl;

[0247] Each R b Choose independently from the following groups: H and C 1-6 alkyl;

[0248] R c Choose from the following groups: H, C 1-6 Alkyl and C 1-6 Hydroxyalkyl;

[0249] R 4 It is C 2-6 Alkyl, C 3-6 cycloalkyl, C 1-6 hydroxyalkyl, heterocyclic, aryl, or heteroaryl; wherein the heterocyclic, aryl, and heteroaryl groups are optionally surrounded by one or two R groups. 4a Group substitution;

[0250] R 5 It is a heterocyclic, aryl, or heteroaryl group; wherein the heterocyclic, aryl, or heteroaryl group is optionally surrounded by one or two R groups. 5a Group substitution;

[0251] One or two R 4a and R 5a Each of the groups, when present, is independently selected from C. 1-6 Alkyl, C 1-6 Hydroxyalkyl, hydroxyl, halogen, C 1-6 Halogenated alkyl groups and C 1-6 alkoxy; or any two R 4a and R 5a When groups exist on the same carbon atom, they form oxo groups together.

[0252] Where R 1 When it is hydrogen, R 2 Not -C 0-6 Alkylene-SO2R 4 .

[0253] In one embodiment, the compound of formula (I) is the compound of formula (P-2):

[0254]

[0255] Or its pharmaceutically acceptable salt;

[0256] Where R 1 It is hydrogen, C 1-6 Alkyl, heterocyclic, aryl, heteroaryl, -C 0-6 Alkylene-SO2R 4 or -C 0-6 Alkylene-SO2R 5 ; wherein the heterocyclic group, aryl group, and heteroaryl group are optionally surrounded by one or two R groups. 1a Group substitution;

[0257] R 2 It is a heterocyclic group, aryl group, heteroaryl group, -C 0-6 Alkylene-SO2R 4 or -C 0-6 Alkylene-SO2R 5 ; wherein the heterocyclic group, aryl group, and heteroaryl group are optionally surrounded by one or two R groups. 2a Group substitution;

[0258] Or R 1 and R 2 Together with the attached nitrogen, it forms an optional structure with one or two R 1a N-linked heteroaryl groups or N-linked heterocyclic groups with substituent groups;

[0259] R 3 Choose from the following groups: - OR a 3-6 membered heterocyclic groups and 3-6 membered heteroaryl groups;

[0260] R a Choose from the following groups: H, -P(O)(R b )2、-C(O)R c -C(O)OR c C 1-6 Alkyl and C 1-6 Hydroxyalkyl;

[0261] Each R b Choose independently from the following groups: H and C 1-6 alkyl;

[0262] R c Choose from the following groups: H, C 1-6 Alkyl and C 1-6 Hydroxyalkyl;

[0263] R 4 It is C 2-6 Alkyl, C 3-6 cycloalkyl, C 1-6 hydroxyalkyl, heterocyclic, aryl, or heteroaryl; wherein the heterocyclic, aryl, and heteroaryl groups are optionally surrounded by one or two R groups. 4a Group substitution; and

[0264] R 5 It is a heterocyclic, aryl, or heteroaryl group; wherein the heterocyclic, aryl, or heteroaryl group is optionally surrounded by one or two R groups. 5a Group substitution;

[0265] One or two R 1a R 2a R 4a and R 5a Each of the groups, when present, is independently selected from C. 1-6 Alkyl, C 1-6 Hydroxyalkyl, hydroxyl, halogen, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 Haloalkoxy and cyano; or any two R 1a R 2a R 4a and R 5a When groups exist on the same carbon atom, they form oxo groups together.

[0266] Where R 1 When it is hydrogen, R 2 Not -C0-6 Alkylene-SO2R 4 .

[0267] In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), R 3 Yes - OR a In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), R 3 It is -OH. In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), R 3 It is -OP(O)(R b 2. In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), R 3 It is -OP(O)H2. In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), R 3 It is -OP(O)(C 1-6 Alkyl)2. In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), R 3 It is -OP(O)(Me)2. In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), R 3 It is -OP(O)(C 1-6 Alkyl group (H). In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), R 3 It is -OP(O)(Me)(H). In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), R 3 It is -C(O)R c In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), R 3 It is -C(O)H. In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), R 3 It is -C(O)C 1-6Alkyl group. In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), R 3 It is -C(O)C 1-6 Hydroxyalkyl. In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), R 3 It is -C(O)OR c In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), R 3 It is -COOH. In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), R 3 It is -C(O)OC 1-6 Alkyl group. In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), R 3 It is -C(O)OC 1-6 Hydroxyalkyl. In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), R 3 Yes - OC 1-6 Alkyl group. In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), R 3 Yes - OC 1-6 Hydroxyalkyl.

[0268] In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), R 3 It is a 3-6 membered heterocyclic group. In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), R 3 It is a 3-6 membered heterocyclic group containing one, two, or three atoms selected from nitrogen, oxygen, and sulfur. In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), R 3 It is a 3-6 membered heterocyclic group containing one nitrogen atom. In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), R 3 It is a 3-6 membered heterocyclic group containing two or three nitrogen atoms.

[0269] In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), R 3 It is a 3-6 membered heteroaryl group. In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), R 3 It is a 3-6 membered heteroaryl group containing one, two, three, or four atoms selected from nitrogen, oxygen, and sulfur atoms. In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), R 3 It is a 3-6 membered heteroaryl group containing one nitrogen atom. In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), R 3 It is a 3-6 membered heteroaryl group containing two or three nitrogen atoms. In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), R 3 It is a 3-6 membered heteroaryl group containing four nitrogen atoms. In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), R 3 It is a tetrazolium or triazole. In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), R 3 yes in It is the attachment point to the rest of the compound.

[0270] In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), including any of the foregoing, R 1 It is hydrogen. In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), including any of the foregoing, R 1 It is C 1-6 Alkyl group. In one embodiment of formula (I), (PI), or (P-2), comprising any of the foregoing, R 1 It is a heterocyclic, aryl, or heteroaryl group. In one embodiment of formula (I), (PI), or (P-2), including any of the foregoing, R 1 It is a heterocyclic, aryl, or heteroaryl group. In one embodiment of formula (I), (PI), or (P-2), including any of the foregoing, R 1 It is -C 0-6 Alkylene-SO2R 4or -C 0-6 Alkylene-SO2R 5 .

[0271] In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), including any of the foregoing, R 1 It is C 1-6 Alkyl, wherein C 1-6 The alkyl group is unsubstituted. In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), including any of the foregoing, R 1 It is C 1-6 Alkyl, wherein C 1-6 Alkyl groups are formed by one or two R groups. 2a Group substitution. In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), including any of the foregoing, R 1 It is C 1-6 Alkyl, wherein C 1-6 Alkyl groups are C 1-6 Hydroxyalkyl substitution. In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), including any of the foregoing, R 1 yes in It is the attachment point to the rest of the compound. In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), including any of the foregoing, R 1 It is C 1-6 Alkyl, wherein C 1-6 Alkyl groups are C 1-6 Alkyl substitution. In one embodiment of formula (I), (Ia)-(Ic), (P-1), (P-2), or (P-Ia)-(P-Ic), including any of the foregoing, R 1 yes in It is the attachment point to the rest of the compound.

[0272] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 2 It is a heterocyclic group, wherein the heterocyclic group is unsubstituted. In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 2 It is a heterocyclic group, wherein the heterocyclic group is separated by one or two R2a Group substitution. In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 2 It is a 5-12 membered monocyclic or bicyclic heterocyclic group containing one, two, or three atoms selected from nitrogen, oxygen, and sulfur, wherein, when nitrogen is present, the nitrogen is optionally converted to C 1-6 Alkyl or C 3-6 Cycloalkyl substitution, wherein the heterocyclic group is optionally replaced by one or two R groups. 2a Group substitution.

[0273] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 2 It is an aryl group, wherein the aryl group is unsubstituted. In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 2 It is a 5-12 member monocyclic or bicyclic aryl group, wherein the aryl group is optionally surrounded by one or two R groups. 2a Group substitution. In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 2 It is an unsubstituted phenyl group. In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 2 Is it by one or two Rs 2a A phenyl group substituted with a radical. In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), comprising any of the foregoing, R 2 It is selected from halogen, C 1-6 Alkoxy, C 1-6 Alkyl and C 1-6 R of haloalkyl 2a A phenyl group substituted with a radical.

[0274] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 2 It is a heteroaryl group, wherein the heteroaryl group is unsubstituted. In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 2 It is a heteroaryl group, wherein the heteroaryl group is optionally surrounded by one or two R groups. 2a Group substitution. In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 2It is a 5-12 membered monocyclic or bicyclic heteroaryl group containing one, two, three, or four atoms selected from nitrogen, oxygen, and sulfur, wherein, when nitrogen is present, the nitrogen is optionally converted to C 1-6 Alkyl or C 3-6 Cycloalkyl substitution, wherein the heteroaryl group is optionally replaced by one or two R groups. 2a Group substitution.

[0275] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 2 It is arbitrarily C 1-6 Alkyl or C 3-6 Cycloalkyl-substituted 5-12 membered monocyclic heteroaryl groups containing one nitrogen atom, wherein the heteroaryl group is optionally replaced by two R groups on the same carbon atom. 2a Group substitution, the two R 2a Groups link together to form oxo groups.

[0276] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 2 Selected from in It is the attachment point to the rest of the compound.

[0277] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 2 Selected from in It is the attachment point to the rest of the compound; and R 2a As defined in this article.

[0278] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 2a Selected from C 1-6 Alkyl, C 1-6 Halogenated alkyl groups, halogens, C 1-6 Alkoxy, cyano and C 1-6 Haloalkoxy. In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 2a It is C 1-6 Hydroxyalkyl or C 1-6 Alkyl group.

[0279] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 2 Selected from

[0280] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 2 Selected from

[0281] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 2 Selected from in It is the attachment point to the rest of the compound.

[0282] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 2 Selected from in It is the attachment point to the rest of the compound.

[0283] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 1 It is hydrogen and R 2 Selected from in It is the attachment point to the rest of the compound.

[0284] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 1 It is methyl and R 2 Selected from in It is the attachment point to the rest of the compound.

[0285] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 1 It is hydrogen and R 2 Selected from in It is the attachment point to the rest of the compound.

[0286] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 1 It is methyl and R 2 Selected from in It is the attachment point to the rest of the compound.

[0287] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 1 It is hydrogen and R 2 Selected from in It is the attachment point to the rest of the compound.

[0288] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 1 It is C 1-6 Alkyl, wherein C 1-6 Alkyl groups are either unsubstituted or C-substituted. 1-6 Hydroxyalkyl or C 1-6 Alkoxy substitution, and R 2 yes in It is the attachment point to the rest of the compound.

[0289] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 1 It is hydrogen; R 2 Selected from Furthermore, R 3 It is OH; among which It is the attachment point to the rest of the compound.

[0290] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 1 It is methyl; R 2 Selected from Furthermore, R 3 It is OH; among which It is the attachment point to the rest of the compound.

[0291] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 1 It is hydrogen; R 2 Selected from Furthermore, R 3 It is OH; among which It is the attachment point to the rest of the compound.

[0292] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 1 It is methyl; R 2 Selected from Furthermore, R 3 It is OH; among which It is the attachment point to the rest of the compound.

[0293] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 1 It is hydrogen; R 2 Selected from Furthermore, R 3 It is -OH, -OP(O)(R b )2 or containing one, two, three or four 3-6 membered heteroaryl groups selected from nitrogen, oxygen and sulfur atoms; wherein It is the attachment point to the rest of the compound.

[0294] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 1 It is hydrogen; R 2 Selected from Furthermore, R 3 It is -OH, -OP(O)(Me)2 or in It is the attachment point to the rest of the compound.

[0295] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 1 It is methyl; R 2 Selected from Furthermore, R 3 It is -OH, -OP(O)(R b )2 or containing one, two, three or four 3-6 membered heteroaryl groups selected from nitrogen, oxygen and sulfur atoms; wherein It is the attachment point to the rest of the compound.

[0296] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 1 It is methyl; R2 Selected from Furthermore, R 3 It is -OH, -OP(O)(Me)2 or in It is the attachment point to the rest of the compound.

[0297] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 1 It is hydrogen; R 2 Selected from Furthermore, R 3 It is -OH, -OP(O)(R b )2 or containing one, two, three or four 3-6 membered heteroaryl groups selected from nitrogen, oxygen and sulfur atoms; wherein It is the attachment point to the rest of the compound.

[0298] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 1 It is hydrogen; R 2 Selected from Furthermore, R 3 It is -OH, -OP(O)(Me)2 or in It is the attachment point to the rest of the compound.

[0299] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 1 It is methyl; R 2 Selected from Furthermore, R 3 It is -OH, -OP(O)(R b )2 or containing one, two, three or four 3-6 membered heteroaryl groups selected from nitrogen, oxygen and sulfur atoms; wherein It is the attachment point to the rest of the compound.

[0300] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 1 It is methyl; R 2 Selected from Furthermore, R 3 It is -OH, -OP(O)(Me)2 or in It is the attachment point to the rest of the compound.

[0301] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 1 It is hydrogen; R 2 Selected from Furthermore, R 3 It is -OH, where It is the attachment point to the rest of the compound.

[0302] In one embodiment of formula (I), (PI), (Ia), (P-Ia), or (P-2), including any of the foregoing, R 1 It is C 1-6 Alkyl, wherein C 1-6 Alkyl groups are either unsubstituted or C-substituted. 1-6 Hydroxyalkyl or C 1-6 Alkyl substitution; R 2 yes Furthermore, R 3 It is -OH, where It is the attachment point to the rest of the compound.

[0303] In one embodiment of formula (I), (PI), (Ib), (P-Ib) or (P-2), including any of the foregoing, R 1 and R 2 Linked together to form N-linked 5-12 membered monocyclic or bicyclic heteroaryl groups containing one, two, three, or four atoms selected from nitrogen, oxygen, and sulfur, wherein the heteroaryl groups are unsubstituted. In one embodiment of formula (I), (PI), (Ib), (P-Ib), or (P-2), including any of the foregoing, R 1 and R 2 They are linked together to form a structure containing one, two, three, or four atoms selected from nitrogen, oxygen, and sulfur (including R). 1 and R 2 The attached nitrogen is an N-linked 5-12 membered monocyclic or bicyclic heteroaryl group, wherein the heteroaryl group is optionally surrounded by one or two R groups. 1a Group substitution. In one embodiment of formula (I), (PI), (Ib), (P-Ib) or (P-2), including any of the foregoing, R 1 and R 2 They are linked together to form a structure containing one, two, three, or four atoms selected from nitrogen, oxygen, and sulfur (including R). 1 and R 2 The attached nitrogen is an N-linked 5-12 membered monocyclic or bicyclic heteroaryl group, wherein the heteroaryl group is optionally connected to an R1a Group substitution. In one embodiment of formula (I), (PI), (Ib), (P-Ib) or (P-2), including any of the foregoing, R 1 and R 2 They are linked together to form a structure containing one, two, three, or four nitrogen atoms (including R). 1 and R 2 The attached nitrogen is an N-linked 5-12 membered monocyclic or bicyclic heteroaryl group, wherein the heteroaryl group is optionally surrounded by one or two R groups. 1a Group substitution.

[0304] In one embodiment of formula (I), (PI), (Ib), (P-Ib) or (P-2), including any of the foregoing, R 1 and R 2 Linked together to form an N-linked 5-12 member monocyclic or bicyclic heterocyclic group containing one, two, three, or four atoms selected from nitrogen, oxygen, and sulfur, wherein the heterocyclic group is unsubstituted. In one embodiment of formula (I), (PI), (Ib), (P-Ib), or (P-2), including any of the foregoing, R 1 and R 2 They are linked together to form a structure containing one, two, three, or four atoms selected from nitrogen, oxygen, and sulfur (including R). 1 and R 2 The attached nitrogen is an N-linked 5-12 member monocyclic or bicyclic heterocyclic group, wherein the heterocyclic group is optionally surrounded by one or two R... 1a Group substitution. In one embodiment of formula (I), (PI), (Ib), (P-Ib) or (P-2), including any of the foregoing, R 1 and R 2 They are linked together to form a structure containing one, two, three, or four atoms selected from nitrogen, oxygen, and sulfur (including R). 1 and R 2 The attached nitrogen is an N-linked 5-12 member monocyclic or bicyclic heterocyclic group, wherein the heterocyclic group is optionally surrounded by an R 1a Group substitution. In one embodiment of formula (I), (PI), (Ib), (P-Ib) or (P-2), including any of the foregoing, R 1 and R 2 They are linked together to form a structure containing one, two, three, or four nitrogen atoms (including R). 1 and R 2 The attached nitrogen is an N-linked 5-12 member monocyclic or bicyclic heterocyclic group, wherein the heterocyclic group is optionally surrounded by one or two R... 1aGroup substitution. In one embodiment of formula (I), (PI), (Ib), (P-Ib) or (P-2), including any of the foregoing, R 1 and R 2 They connect together to form a structure containing one nitrogen atom and S(O). 0-2 The N-linked 5-12 membered monocyclic or bicyclic heterocyclic group, wherein the nitrogen atom is R 1 and R 2 The attached nitrogen, wherein the heterocyclic group is optionally surrounded by one or two R 1a Group substitution. In one embodiment of formula (I), (PI), (Ib), (P-Ib) or (P-2), including any of the foregoing, R 1 and R 2 They are linked together to form a 5-12 membered monocyclic or bicyclic heterocyclic group containing a nitrogen atom and an N-linked S(O)2, wherein the nitrogen atom is R 1 and R 2 The attached nitrogen, wherein the heterocyclic group is optionally surrounded by one or two R 1a Group substitution.

[0305] In one embodiment of formula (I), (PI), (Ib), (P-Ib) or (P-2), including any of the foregoing, R 1 and R 2 Connected together to form in It is the attachment point to the rest of the compound.

[0306] In one embodiment of formula (I), (PI), (Ib), (P-Ib) or (P-2), including any of the foregoing, R 1 and R 2 Connected together to form in It is the attachment point to the rest of the compound.

[0307] In one embodiment of formula (I), (PI), (Ib), (P-Ib) or (P-2), including any of the foregoing, R 1 and R 2 Connected together to form in It is the attachment point to the rest of the compound.

[0308] In one embodiment of formula (I), (PI), (Ib), (P-Ib) or (P-2), including any of the foregoing, R 1 and R 2 Connected together to form And R 3It is OH; among which It is the attachment point to the rest of the compound.

[0309] In one embodiment of formula (I), (PI), (Ib), (P-Ib) or (P-2), including any of the foregoing, R 1 and R 2 Connected together to form Furthermore, R 3 It is -OH, -OP(O)(R b )2 or containing one, two, three or four 3-6 membered heteroaryl groups selected from nitrogen, oxygen and sulfur atoms; wherein It is the attachment point to the rest of the compound.

[0310] In one embodiment of formula (I), (PI), (Ib), (P-Ib) or (P-2), including any of the foregoing, R 1 and R 2 Connected together to form Furthermore, R 3 It is -OH, -OP(O)(Me)2 in It is the attachment point to the rest of the compound.

[0311] In one embodiment of formula (I), (PI), (Ic), (P-Ic) or (P-2), including any of the foregoing, R 2 It is -C 0-6 Alkylene-SO2R 4 In one embodiment of formula (I), (PI), (Ic), (P-Ic), or (P-2), R 2 It is -C 0-6 Alkylene-SO2R 5 .

[0312] In one embodiment of formula (I), (PI), (Ic), (P-Ic) or (P-2), including any of the foregoing, R 2 It is -C 0-6 Alkylene-SO2R 4 And R 4 It is C 2-6 Alkyl group. In one embodiment of formula (I), (PI), (Ic), (P-Ic), or (P-2), comprising any of the foregoing, R 2 It is -C 0-6 Alkylene-SO2R 4 And R 4 It is C 3-6Cycloalkyl. In one embodiment of formula (I), (PI), (Ic), (P-Ic), or (P-2), comprising any of the foregoing, R 2 It is -C 0-6 Alkylene-SO2R 4 And R 4 It is C 1-6 Hydroxyalkyl. In one embodiment of formula (I), (PI), (Ic), (P-Ic), or (P-2), comprising any of the foregoing, R 2 It is -C 0-6 Alkylene-SO2R 4 And R 4 It is a heterocyclic group, and the heterocyclic group is optionally separated by one or two R... 4a Group substitution. In one embodiment of formula (I), (PI), (Ic), (P-Ic), or (P-2), including any of the foregoing, R 2 It is -C 0-6 Alkylene-SO2R 4 And R 4 It is aryl, and the aryl group is optionally separated by one or two R groups. 4a Group substitution. In one embodiment of formula (I), (PI), (Ic), (P-Ic), or (P-2), including any of the foregoing, R 2 It is -C 0-6 Alkylene-SO2R 4 And R 4 It is a heteroaryl group, and the heteroaryl group is optionally surrounded by one or two R groups. 4a Group substitution.

[0313] In one embodiment of formula (I), (PI), (Ic), (P-Ic) or (P-2), including any of the foregoing, R 2 It is -C 0-6 Alkylene-SO2R 5 And R 5 It is a heterocyclic group, and the heterocyclic group is optionally separated by one or two R... 5a Group substitution. In one embodiment of formula (I), (PI), (Ic), (P-Ic), or (P-2), including any of the foregoing, R 2 It is -C 0-6 Alkylene-SO2R 5 And R 5 It is aryl, and the aryl group is optionally separated by one or two R groups. 5a Group substitution. In one embodiment of formula (I), (PI), (Ic), (P-Ic), or (P-2), including any of the foregoing, R 2 It is -C0-6 Alkylene-SO2R 5 And R 5 It is a heteroaryl group, and the heteroaryl group is optionally surrounded by one or two R groups. 5a Group substitution.

[0314] In one embodiment of formula (I), (PI), (Ic), (P-Ic) or (P-2), including any of the foregoing, R 2 It is -C 0-6 Alkylene-SO2R 5 And R 5 It is a 5-12 membered monocyclic heteroaryl group containing two atoms selected from nitrogen and oxygen, wherein R 5 Optionally by one or two R 5a Group substitution. In one embodiment of formula (I), (PI), (Ic), (P-Ic), or (P-2), including any of the foregoing, R 5 Selected from

[0315] In one embodiment of formula (I), (PI), (Ic), (P-Ic) or (P-2), including any of the foregoing, R 1 It is methyl and R 2 It is -C 0-6 Alkylene-SO2R 4 In one embodiment of formula (I), (PI), (Ic), (P-Ic), or (P-2), including any of the foregoing, R 1 It is hydrogen and R 2 It is -C 0-6 Alkylene-SO2R 5 In one embodiment of formula (I), (PI), (Ic), (P-Ic), or (P-2), including any of the foregoing, R 1 It is methyl and R 2 It is -C 0-6 Alkylene-SO2R 5 .

[0316] In one embodiment of formula (I), (PI), (Ic), (P-Ic) or (P-2), including any of the foregoing, R 2 Selected from in It is the attachment point to the rest of the compound.

[0317] In one embodiment of formula (I), (PI), (Ic), (P-Ic) or (P-2), including any of the foregoing, R 1 It is hydrogen, R2 Selected from And R 3 It is -OH; where It is the attachment point to the rest of the compound.

[0318] In one embodiment of formula (I), (PI), (Ic), (P-Ic) or (P-2), including any of the foregoing, R 1 It is methyl, R 2 Selected from And R 3 It is -OH; where It is the attachment point to the rest of the compound.

[0319] In one embodiment of formula (I), (PI), (Ic), (P-Ic) or (P-2), including any of the foregoing, R 2 Selected from And R 3 It is -OH; where It is the attachment point to the rest of the compound.

[0320] In one embodiment of formula (I), (PI), (Ic), (P-Ic) or (P-2), including any of the foregoing, R 1 It is methyl; R 2 It is -C 0-6 Alkylene-SO2R 4 And R 3 It is -OH, -OP(O)(R b )2 or a 3-6 membered heteroaryl group containing one, two, three, or four atoms selected from nitrogen, oxygen, and sulfur atoms. In one embodiment of formula (I), (PI), (Ic), (P-Ic), or (P-2), including any of the foregoing, R 1 It is hydrogen; R 2 It is -C 0-6 Alkylene-SO2R 5 And R 3 It is -OH, -OP(O)(R b )2 or a 3-6 membered heteroaryl group containing one, two, three, or four atoms selected from nitrogen, oxygen, and sulfur atoms. In one embodiment of formula (I), (Ic), or (P-2), including any of the foregoing, R 1 It is methyl; R 2 It is -C 0-6 Alkylene-SO2R 5 And R 3 It is -OH, -OP(O)(Rb )2 or 3-6 heteroaryl groups containing one, two, three or four atoms selected from nitrogen, oxygen and sulfur atoms.

[0321] In one embodiment of formula (I), (PI), (Ic), (P-Ic) or (P-2), including any of the foregoing, R 1 It is methyl; R 2 It is -C 0-6 Alkylene-SO2R 4 Furthermore, R 3 It is -OH, -OP(O)(Me)2 or in It is the attachment point to the rest of the compound. In one embodiment of formula (I), (Ic), or (P-2), including any of the foregoing, R 1 It is hydrogen; R 2 It is -C 0-6 Alkylene-SO2R 5 And R 3 It is -OH, -OP(O)(Me)2 or in It is the attachment point to the rest of the compound. In one embodiment of formula (I), (PI), (Ic), (P-Ic), or (P-2), including any of the foregoing, R 1 It is methyl; R 2 It is -C 0-6 Alkylene-SO2R 5 Furthermore, R 3 It is -OH, -OP(O)(Me)2 or in It is the attachment point to the rest of the compound.

[0322] In one embodiment of formula (I), (PI), (Ic), (P-Ic) or (P-2), including any of the foregoing, R 1 It is hydrogen, R 2 Selected from And R 3 It is -OH, -OP(O)(R b )2 or containing one, two, three or four 3-6 membered heteroaryl groups selected from nitrogen, oxygen and sulfur atoms; wherein It is the attachment point to the rest of the compound.

[0323] In one embodiment of formula (I), (PI), (Ic), (P-Ic) or (P-2), including any of the foregoing, R 1 It is methyl, R 2 Selected from And R 3 It is -OH, -OP(O)(R b )2 or containing one, two, three or four 3-6 membered heteroaryl groups selected from nitrogen, oxygen and sulfur atoms; wherein It is the attachment point to the rest of the compound.

[0324] In one embodiment of formula (I), (PI), (Ic), (P-Ic) or (P-2), including any of the foregoing, R 1 It is hydrogen, R 2 Selected from And R 3 It is -OH, -OP(O)(Me)2 or in It is the attachment point to the rest of the compound.

[0325] In one embodiment of formula (I), (PI), (Ic), (P-Ic) or (P-2), including any of the foregoing, R 1 It is methyl, R 2 Selected from And R 3 It is -OH, -OP(O)(Me)2 or in It is the attachment point to the rest of the compound.

[0326] In one embodiment of formula (I), (PI), (Ic), (P-Ic) or (P-2), including any of the foregoing, R 4 Selected from in It is the attachment point to the rest of the compound.

[0327] In one embodiment of formula (I), (PI), (Ic), (P-Ic) or (P-2), including any of the foregoing, R 4 Selected from in It is the attachment point to the rest of the compound; and R 4a As defined in this article.

[0328] In one embodiment of formula (I), (PI), (Ic), (P-Ic) or (P-2), including any of the foregoing, R 4 Selected from in It is the attachment point to the rest of the compound.

[0329] In one embodiment of formula (I), (PI), (Ic), (P-Ic) or (P-2), including any of the foregoing, R 5 Selected from in It is the attachment point to the rest of the compound.

[0330] In one embodiment of formula (I), (PI), (Ic), (P-Ic) or (P-2), including any of the foregoing, R 5 Selected from in It is the attachment point to the rest of the compound; and R 5a As defined in this article.

[0331] In one embodiment of formula (I), (PI), (Ic), (P-Ic) or (P-2), including any of the foregoing, R 4 Selected from

[0332] in It is the attachment point to the rest of the compound.

[0333] In one embodiment, the compound of formula (I) has the following formula:

[0334]

[0335] Or a pharmaceutically acceptable salt thereof; wherein R 1 R 2 and R 3 As described in any implementation of formula (I) herein.

[0336] Non-limiting examples of compounds of formula (Ia) include:

[0337]

[0338] Or a pharmaceutically acceptable salt thereof; wherein R 2 and R b As described in any embodiment of formula (Ia) herein. Non-limiting examples of compounds of formula (Ib) include:

[0339]

[0340] Or a pharmaceutically acceptable salt thereof; wherein R 1 R 2 and R b As described in any implementation of formula (Ib) as described herein.

[0341] Non-limiting examples of compounds of formula (Ic) include:

[0342]

[0343] Or a pharmaceutically acceptable salt thereof; wherein R 2 and R b As described in any implementation of formula (Ic) as described herein.

[0344] In some embodiments, the compounds of formula (P-2), formula (PI), formula (P-Ia), formula (P-Ib), formula (P-Ic), formula (I), formula (Ia), formula (Ib), or formula (Ic) have zero de or %de. In some embodiments, the compounds of formula (P-2), formula (PI), formula (P-Ia), formula (P-Ib), formula (P-Ic), formula (I), formula (Ia), formula (Ib), or formula (Ic) have a de or %de greater than zero. For example, in some embodiments, the compounds of formula (P-2), formula (PI), formula (P-Ia), formula (P-Ib), formula (P-Ic), formula (I), formula (Ia), formula (Ib), or formula (Ic) have about 10 de or %de. In some embodiments, the compound of formula (P-2), formula (PI), formula (P-Ia), formula (P-Ib), formula (P-Ic), formula (I), formula (Ia), formula (Ib), or formula (Ic) has about 25% de or % de. In some embodiments, the compound of formula (P-2), formula (PI), formula (P-Ia), formula (P-Ib), formula (P-Ic), formula (I), formula (Ia), formula (Ib), or formula (Ic) has about 50% de or % de. In some embodiments, the compound of formula (P-2), formula (PI), formula (P-Ia), formula (P-Ib), formula (P-Ic), formula (I), formula (Ia), formula (Ib), or formula (Ic) has about 75% de or % de. In some embodiments, the compounds of formula (P-2), formula (PI), formula (P-Ia), formula (P-Ib), formula (P-Ic), formula (I), formula (Ia), formula (Ib), or formula (Ic) have about 80% de or % de. In some embodiments, the compounds of formula (P-2), formula (PI), formula (P-Ia), formula (P-Ib), formula (P-Ic), formula (I), formula (Ia), formula (Ib), or formula (Ic) have about 85% de or % de. In some embodiments, the compounds of formula (P-2), formula (PI), formula (P-Ia), formula (P-Ib), formula (P-Ic), formula (I), formula (Ia), formula (Ib), or formula (Ic) have about 90% de or % de. In some embodiments, the compounds of formula (P-2), formula (PI), formula (P-Ia), formula (P-Ib), formula (P-Ic), formula (I), formula (Ia), formula (Ib), or formula (Ic) have about 95% de or % de. In some embodiments, the compounds of formula (P-2), formula (PI), formula (P-Ia), formula (P-Ib), formula (P-Ic), formula (I), formula (Ia), formula (Ib), or formula (Ic) have about 97% de or % de.In some embodiments, the compounds of formula (P-2), formula (PI), formula (P-Ia), formula (P-Ib), formula (P-Ic), formula (I), formula (Ia), formula (Ib), or formula (Ic) have about 98% de or % de. In some embodiments, the compounds of formula (P-2), formula (PI), formula (P-Ia), formula (P-Ib), formula (P-Ic), formula (P-2), formula (I), formula (Ia), formula (Ib), or formula (Ic) have about 99% de or % de. In some embodiments, the compounds described herein have 100% de or % de.

[0345] In one embodiment, the compound of formula (P-2), formula (PI), formula (P-Ia), formula (P-Ib), formula (P-Ic), formula (I), formula (Ia), formula (Ib), or formula (Ic), or a pharmaceutically acceptable salt thereof, is at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 97%, at least about 99%, or even 100% of the C-16(R)-epomer by weight, based solely on the weight of the compound of formula (P-2), formula (PI), formula (P-Ia), formula (P-Ib), formula (P-Ic), formula (I), formula (Ia), formula (Ib), or formula (Ic) (i.e., if the compound is present as a pharmaceutically acceptable salt, the weight of the pharmaceutically acceptable salt is not included). In one embodiment, the compound of formula (P-2), formula (PI), formula (P-Ia), formula (P-Ib), formula (P-Ic), formula (I), (Ia), (Ib), or (Ic), or a pharmaceutically acceptable salt thereof, is about 85%-95% (R)-C-16 epimer. In one embodiment, the compound of formula (P-2), formula (PI), formula (P-Ia), formula (P-Ib), formula (P-Ic), formula (I), (Ia), (Ib), or (Ic), or a pharmaceutically acceptable salt thereof, is about 90%-95% (R)-C-16 epimer.

[0346] In one embodiment, the compound of formula (P-2), formula (PI), formula (P-Ia), formula (P-Ib), formula (P-Ic), formula (I), formula (Ia), formula (Ib), or formula (Ic), or a pharmaceutically acceptable salt thereof, is 50% by weight of the C-16(S)-epomer and 50% by weight of the C-16(R)-epomer, based solely on the weight of the compound of formula (P-2), formula (PI), formula (P-Ia), formula (P-Ib), formula (P-Ic), formula (I), formula (Ia), formula (Ib), or formula (Ic) (i.e., if the compound is present as a pharmaceutically acceptable salt, the weight of the pharmaceutically acceptable salt is not included).

[0347] In one embodiment, the compound of formula (P-2), formula (PI), formula (P-Ia), formula (P-Ib), formula (P-Ic), formula (I), formula (Ia), formula (Ib), or formula (Ic), or a pharmaceutically acceptable salt thereof, is at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 97%, at least about 99%, or even 100% of the C-16(S)-epomer by weight, based solely on the weight of the compound of formula (P-2), formula (PI), formula (P-Ia), formula (P-Ib), formula (P-Ic), formula (I), formula (Ia), formula (Ib), or formula (Ic) (i.e., if the compound is present as a pharmaceutically acceptable salt, the weight of the pharmaceutically acceptable salt is not included). In one embodiment, the compound of formula (P-2), formula (PI), formula (P-Ia), formula (P-Ib), formula (P-Ic), formula (I), (Ia), (Ib), or (Ic), or a pharmaceutically acceptable salt thereof, is about 85%-95% (S)-C-16 epimer. In one embodiment, the compound of formula (P-2), formula (PI), formula (P-Ia), formula (P-Ib), formula (P-Ic), formula (I), (Ia), (Ib), or (Ic), or a pharmaceutically acceptable salt thereof, is about 90%-95% (S)-C-16 epimer.

[0348] In some embodiments, the compounds and pharmaceutical compositions disclosed herein for use in treating disorders or diseases mediated by the mTOR pathway are more selective in binding FKBP12 than other proteins in the FK506 binding protein (FKBP) group.

[0349] In some other embodiments, the compounds and pharmaceutical compositions disclosed herein for use in the treatment of disorders or diseases mediated by the mTOR pathway exhibit unusual and surprising pharmacokinetic profiles and enhanced pharmacodynamic selectivity in the target FKBP. These compounds and pharmaceutical compositions may be used to treat age-related or aging-related diseases, diabetic complications, cancer, and inflammation-related disorders.

[0350] In some other embodiments, the compounds and pharmaceutical compositions disclosed herein for use in the treatment of disorders or diseases mediated by the mTOR pathway, wherein the compounds inhibit S6K1 phosphorylation in FKBP12 KO cells at least twice as efficiently as the rapamycin analog RAD001 compared to cells expressing FKBP12.

[0351] In some other embodiments, the compounds and pharmaceutical compositions disclosed herein for use in the treatment of disorders or diseases mediated by the mTOR pathway, wherein the compounds inhibit S6K1 phosphorylation in FKBP12 KO cells at least 10-fold less efficiently than the rapamycin analog RAD001 compared to cells expressing FKBP12.

[0352] In some other embodiments, the compounds and pharmaceutical compositions disclosed herein for use in the treatment of disorders or diseases mediated by the mTOR pathway, wherein the compounds inhibit S6K1 phosphorylation in FKBP12 KO cells at least 100-fold less efficiently than the rapamycin analog RAD001 compared to cells expressing FKBP12.

[0353] In some other embodiments, the compounds and pharmaceutical compositions disclosed herein for use in the treatment of disorders or diseases mediated by the mTOR pathway, wherein the compounds require approximately 10-fold higher concentrations in FKBP12 KO cells compared to cells expressing FKBP12, and compared to the rapamycin analog RAD001, to achieve 20% inhibition of S6K1 cell signaling.

[0354] In some other embodiments, the compounds and pharmaceutical compositions disclosed herein for use in the treatment of disorders or diseases mediated by the mTOR pathway, wherein the compounds require approximately 20-fold higher concentrations in FKBP12 KO cells compared to cells expressing FKBP12, and compared to the rapamycin analog RAD001, to achieve 20% inhibition of S6K1 cell signaling.

[0355] In some other embodiments, the compounds and pharmaceutical compositions disclosed herein for use in the treatment of disorders or diseases mediated by the mTOR pathway, wherein the compounds require approximately 100-fold higher concentrations in FKBP12 KO cells compared to cells expressing FKBP12, and compared to the rapamycin analog RAD001, to achieve 20% inhibition of S6K1 cell signaling.

[0356] In some other embodiments, the compounds and pharmaceutical compositions disclosed herein for use in the treatment of disorders or diseases mediated by the mTOR pathway, wherein the compounds require approximately 500-fold higher concentrations in FKBP12 KO cells compared to cells expressing FKBP12, and compared to the rapamycin analog RAD001, to achieve 20% inhibition of S6K1 cell signaling.

[0357] In some other embodiments, the compounds and pharmaceutical compositions disclosed herein for use in the treatment of disorders or diseases mediated by the mTOR pathway, wherein the compounds require approximately 1000 times higher concentrations in FKBP12 KO cells compared to cells expressing FKBP12, and compared to the rapamycin analog RAD001, to achieve 20% inhibition of S6K1 cell signaling.

[0358] In some other embodiments, the “FKBP12 selective rapamycin analog” is a rapamycin analog that, while effective in assays using wild-type cells, is about 10 to about 1000 times less potent than RAD001 in cell lines that do not express FKBP12 (e.g., FKBP12 knockout cells). As used in this context, “potency” can be expressed as the concentration of rapamycin analog required to achieve 20% inhibition of S6K1(Thr389) phosphorylation in a cell-based assay, such as the one used in Example 3 and Figures 1-11. For example, if a rapamycin analog inhibits approximately 20% of S6K1 (Thr389) phosphorylation in an assay using an FKBP12 knockout cell line, and its potency is at least 20x lower (e.g., 25x, 30x, 50x, 100x, 200x, 500x, 1000x, etc.) than the similar inhibition level achieved by RAD001 in the same assay using the same cell line expressing normal FKBP12, then the rapamycin analog is considered to be FKBP12 selective.

[0359] In some other embodiments, the compounds and pharmaceutical compositions disclosed herein for use in the treatment of disorders or diseases mediated by the mTOR pathway, wherein the compounds require approximately 20-fold higher concentrations in FKBP12 KO cells compared to cells expressing FKBP12, and compared to the rapamycin analog RAD001, to achieve 30% inhibition of S6K1 cell signaling.

[0360] In some other embodiments, the compounds and pharmaceutical compositions disclosed herein for use in the treatment of disorders or diseases mediated by the mTOR pathway, wherein the compounds require approximately 100-fold higher concentrations in FKBP12 KO cells compared to cells expressing FKBP12, and compared to the rapamycin analog RAD001, to achieve 30% inhibition of S6K1 cell signaling.

[0361] In some other embodiments, the compounds and pharmaceutical compositions disclosed herein for use in the treatment of disorders or diseases mediated by the mTOR pathway, wherein the compounds require approximately 500-fold higher concentrations in FKBP12 KO cells compared to cells expressing FKBP12, and compared to the rapamycin analog RAD001, to achieve 30% inhibition of S6K1 cell signaling.

[0362] In some other embodiments, the compounds and pharmaceutical compositions disclosed herein for use in the treatment of disorders or diseases mediated by the mTOR pathway, wherein the compounds require approximately 1000 times higher concentrations in FKBP12 KO cells compared to cells expressing FKBP12, and compared to the rapamycin analog RAD001, to achieve 30% inhibition of S6K1 cell signaling.

[0363] In some other embodiments, the “FKBP12 selective rapamycin analog” is a rapamycin analog that, while effective in assays using wild-type cells, is approximately 20 to 1000 times less potent than RAD001 in cell lines that do not express FKBP12 (e.g., FKBP12 knockout cells). As used herein, “potency” can be expressed as the concentration of rapamycin analog required to achieve 30% inhibition of S6K1(Thr389) phosphorylation in cell-based assays such as those used in Example 3 and Figures 1-11. For example, if a rapamycin analog inhibits approximately 30% of S6K1 (Thr389) phosphorylation in an assay using an FKBP12 knockout cell line, and its potency is at least 20x lower (e.g., 25x, 30x, 50x, 100x, 200x, 500x, 1000x, etc.) than the similar inhibition level achieved by RAD001 in the same assay using the same cell line expressing normal FKBP12, then the rapamycin analog is considered to be FKBP12 selective.

[0364] The non-limiting compounds of this invention include:

[0365]

[0366]

[0367]

[0368]

[0369]

[0370]

[0371]

[0372]

[0373]

[0374]

[0375]

[0376]

[0377]

[0378]

[0379]

[0380]

[0381]

[0382]

[0383]

[0384]

[0385]

[0386]

[0387]

[0388]

[0389]

[0390]

[0391]

[0392]

[0393]

[0394] Pharmaceutical compositions and treatment methods

[0395] This document provides methods for treating and preventing diseases, conditions, or disorders, which involve administering a therapeutic or preventative effective amount of one or more compounds disclosed herein, such as one or more compounds of the formula provided herein. Diseases, disorders, and / or conditions include, but are not limited to, those mediated by the mTOR pathway.

[0396] In some embodiments of the methods described herein, multiple doses of the compound described herein (or a pharmaceutical composition comprising a combination of the compound described herein and any other therapeutic agents mentioned herein) may be administered to a subject over a defined time period. The method according to this embodiment of the disclosure includes sequentially administering multiple doses of the compound described herein to a subject. As used herein, “sequentially administering” means administering doses of the compound to the subject at different time points, such as on different days separated by predetermined intervals (e.g., hours, days, weeks, or months). The disclosure includes methods that sequentially administer a single initial dose of the compound described herein to a patient, followed by one or more second doses of the compound, and optionally subsequently, one or more third doses of the compound.

[0397] The terms “initial dose,” “second dose,” and “third dose” refer to the temporal sequence of administration of the compounds described herein. Thus, the “initial dose” is the dose administered at the start of a treatment regimen (also referred to as the “baseline dose”); the “second dose” is the dose administered after the initial dose; and the “third dose” is the dose administered after the second dose. The initial, second, and third doses may each comprise the same amount of the compounds described herein, but may generally differ from each other in terms of frequency of administration. In some embodiments, the amounts of compounds included in the initial, second, and / or third doses vary from one another during the course of treatment (e.g., adjusted up or down as appropriate). In some embodiments, two or more (e.g., two, three, four, or five) doses are administered at the start of a treatment regimen as a “loading dose,” followed by subsequent doses administered on a less frequent basis (e.g., a “maintenance dose”).

[0398] In some exemplary embodiments of this disclosure, each second and / or third dose is administered 1 to 26 weeks immediately following the preceding dose (e.g., 1, 1...). 1 / 2、2、2 1 / 2, 3, 3 1 / 2、4、4 1 / 2, 5, 5 1 / 2, 6, 6 1 / 2、7、7 1 / 2、8、8 1 / 2, 9, 9 1 / 2, 10, 10 1 / 2, 11, 11 1 / 2, 12, 12 1 / 2, 13, 13 1 / 2, 14, 14 1 / 2, 15, 15 1 / 2, 16, 16 1 / 2, 17, 17 1 / 2, 18, 18 1 / 2, 19, 19 1 / 2, 20, 20 1 / 2、21、21 1 / 2、22、22 1 / 2、23、23 1 / 2、24、24 1 / 2、25、25 1 / 2, 26, 26 1 / 2 or more) administered. As used herein, the phrase “immediately preceding dose” means, in a sequence of multiple administrations, the dose of the compound administered to the patient before the next dose immediately following the one in that sequence, without intermediate doses.

[0399] The method according to this disclosure may include administering any number of second and / or third doses of the compound to a patient. For example, in some embodiments, only a single second dose is administered to the patient. In other embodiments, two or more (e.g., two, three, four, five, six, seven, eight, or more) second doses are administered to the patient. Similarly, in some embodiments, only a single third dose is administered to the patient. In other embodiments, two or more (e.g., two, three, four, five, six, seven, eight, or more) third doses are administered to the patient. The administration regimen may be carried out indefinitely throughout the lifetime of a particular subject, or until such treatment is no longer therapeutically necessary or beneficial.

[0400] In embodiments involving multiple second doses, each second dose may be administered at the same frequency as the other second doses. For example, each second dose may be administered to the patient 1 to 2 weeks or 1 to 2 months immediately following the preceding dose. Similarly, in embodiments involving multiple third doses, each third dose may be administered at the same frequency as the other third doses. For example, each third dose may be administered to the patient 2 to 12 weeks immediately following the preceding dose. In some embodiments of this disclosure, the frequency of administration of the second and / or third doses to the patient may vary throughout the treatment regimen. Following a clinical examination, the physician may also adjust the administration frequency during the treatment process based on the individual patient's needs.

[0401] This disclosure includes administration regimens in which two to six loading doses are administered to a patient at a first frequency (e.g., once a week, once every two weeks, once every three weeks, once a month, once every two months, etc.), followed by two or more maintenance doses administered to the patient at a lower frequency. For example, according to this embodiment of the disclosure, if the loading dose is administered once a month, the maintenance dose can be administered to the patient once every six weeks, once every two months, once every three months, etc.

[0402] This disclosure includes pharmaceutical compositions containing compounds described herein (e.g., compounds of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic), or compounds 3-74 or their salts), as well as pharmaceutically acceptable carriers, diluents, and / or excipients. Examples of suitable carriers, diluents, and excipients include, but are not limited to, buffers (e.g., citrate buffers, succinate buffers, acetate buffers, phosphate buffers, lactate buffers, oxalate buffers, etc.) for maintaining a suitable pH of the composition, carrier proteins (e.g., human serum albumin), saline solutions, polyols (e.g., trehalose, sucrose, xylitol, sorbitol, etc.), surfactants (e.g., polysorbate 20, polysorbate 80, polyoxolate, etc.), antimicrobial agents, and antioxidants.

[0403] This disclosure also includes combination therapies comprising: 1) a compound of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic) or a pharmaceutical composition thereof, and 2) an anti-CD40 antibody. In some embodiments, the compound of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic) or a pharmaceutical composition thereof, and the anti-CD40 antibody are administered in separate dosage forms. In some embodiments, the compound of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic) or a pharmaceutical composition thereof, and the anti-CD40 antibody are administered in a combination dosage form.

[0404] In some implementations, this document describes a method for treating a disorder or disease in a subject with a corresponding need, the method comprising administering a compound of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic) or a pharmaceutically acceptable salt thereof.

[0405] In some implementations, this document describes a method for treating a disorder or disease in a subject with a corresponding need, the method comprising administering a combination therapy comprising: 1) a compound of formula (I), (P-1), (P-2), (Ia)-(Ic) or (P-Ia)-(P-Ic) or a pharmaceutical composition thereof, and 2) an anti-CD40 antibody.

[0406] In some embodiments, this document describes a method for treating a subject with an mTOR pathway-mediated disorder or disease, the method comprising administering a compound of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic) or a pharmaceutically acceptable salt thereof. In one embodiment, the target tissue, target organ, or target cell associated with the pathology of the disease or disorder has an FKBP12 level sufficient to inhibit mTORC1.

[0407] In one implementation, the disease or disorder is selected from the following age-related disorders or diseases: sarcopenia; skin atrophy; cherry angioma; seborrheic keratosis; brain atrophy; atherosclerosis; arteriosclerosis; emphysema; osteoporosis; osteoarthritis; hypertension; erectile dysfunction; cataracts; macular degeneration; glaucoma; stroke; cerebrovascular disease (stroke); chronic kidney disease; diabetic nephropathy; impaired liver function; liver fibrosis; autoimmune hepatitis; endometrial hyperplasia; metabolic dysfunction; renal vascular disease; hearing loss; activity deficit; cognitive decline; tendon stiffness; cardiac dysfunction, such as cardiac hypertrophy and / or systolic dysfunction and / or diastolic dysfunction and / or hypertension; cardiac dysfunction leading to decreased ejection fraction; immunosenescence; Parkinson's disease; Alzheimer's disease or its syndromes; cancer; immunosenescence leading to cancer due to decreased immune surveillance; infection due to decreased immune function; chronic obstructive pulmonary disease (COPD); obesity; anosmia; anosmia; arthritis; and type II diabetes.

[0408] In one implementation, the disease or disorder is cancer. In one implementation, the cancer is selected from kidney cancer, renal cell carcinoma, colorectal cancer, uterine sarcoma, endometrial cancer, breast cancer, ovarian cancer, cervical cancer, gastric cancer, fibrosarcoma, pancreatic cancer, liver cancer, melanoma, leukemia, multiple myeloma, nasopharyngeal carcinoma, prostate cancer, lung cancer, glioblastoma, bladder cancer, mesothelioma, head cancer, rhabdomyosarcoma, sarcoma, lymphoma, and cervical cancer.

[0409] In one implementation, the disease or disorder is graft-versus-host disease (GvHD) or its syndrome.

[0410] In one implementation, the disease or disorder is a facial angiofibroma associated with tuberous sclerosis syndrome.

[0411] In one implementation, the disease or disorder is an advanced, unresectable, or metastatic malignant perivascular epithelioid tumor.

[0412] In some implementations, this document also describes methods for inducing immune tolerance and / or preventing transplant rejection in subjects with appropriate need, methods comprising administering compounds of formula (I), (P-1), (Ia)-(Ic), or (P-Ia)-(P-Ic) or pharmaceutically acceptable salts thereof.

[0413] This disclosure provides compounds of formula (I), (P-1), (P-2), (Ia)-(Ic) or (P-Ia)-(P-Ic) or pharmaceutically acceptable salts thereof for the treatment of the diseases and disorders described herein, such as age-related disorders, or diseases and disorders currently approved for treatment with rapamycin analogs (such as RAD001).

[0414] In one aspect, this disclosure provides a method for treating a disorder or disease mediated by the mTOR pathway in a subject with appropriate need, the method comprising administering to the subject a therapeutically effective amount of a compound of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic) or a pharmaceutically acceptable salt thereof.

[0415] In the implementation plan, the efficacy of the treatment is determined based on experience, for example, compared with rapamycin or RAD001.

[0416] In another aspect, this disclosure provides a method for treating a subject with or previously determined to have FKBP12 levels sufficient to inhibit mTORC1, the method comprising administering to the subject in accordance with the need a therapeutically effective amount of a compound of formula (I), (P-1), (P-2), (Ia)-(Ic) or (P-Ia)-(P-Ic), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition or combination of pharmaceuticals described herein.

[0417] In the implementation plan, the disease or disorder is selected from sarcopenia, skin atrophy, cherry angioma, seborrheic keratosis, brain atrophy, atherosclerosis, arteriosclerosis, emphysema, osteoporosis, osteoarthritis, hypertension, erectile dysfunction, cataracts, macular degeneration, glaucoma, stroke, cerebrovascular disease (stroke), chronic kidney disease, diabetic nephropathy, impaired liver function, liver fibrosis, autoimmune hepatitis, endometrial hyperplasia, metabolic dysfunction, renal vascular disease, hearing loss, activity deficit, cognitive decline, tendon stiffness, cardiac dysfunction such as cardiac hypertrophy and / or systolic dysfunction and / or diastolic dysfunction and / or hypertension, cardiac dysfunction leading to decreased ejection fraction, immunosenescence, Parkinson's disease, Alzheimer's disease, cancer, immunosenescence leading to cancer due to decreased immune surveillance, infection due to decreased immune function, chronic obstructive pulmonary disease (COPD), obesity, anosmia, anosmia, arthritis, and type II diabetes (including complications arising from diabetes such as kidney failure, blindness, and neurological disorders).

[0418] In the implementation plan, the disorder is liver fibrosis.

[0419] In another aspect, this disclosure provides a method for treating a disease or disorder in a subject with a corresponding need, the method comprising administering to the subject a therapeutically effective amount of a compound of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic) or a pharmaceutically acceptable salt thereof, a pharmaceutical composition comprising a compound of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a compound of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic) or a pharmaceutically acceptable salt thereof, wherein the disorder or disease is selected from:

[0420] • Acute or chronic organ or tissue graft rejection;

[0421] • Transplant vascular disease;

[0422] • Smooth muscle cell proliferation and migration lead to thickening of the vascular intima, vascular occlusion, obstructive coronary atherosclerosis, and restenosis;

[0423] • Autoimmune diseases and inflammatory conditions;

[0424] ·asthma;

[0425] Multidrug resistance (MDR);

[0426] Fungal infection;

[0427] Inflammation;

[0428] ·Infect;

[0429] Age-related diseases;

[0430] Neurodegenerative diseases;

[0431] • Proliferative disorders, especially cancer;

[0432] • Seizures and seizure-related disorders; and

[0433] • Mitochondrial myopathy and mitochondrial stress.

[0434] In the implementation plan, the disorder is a disorder that includes processes of fibrosis and / or inflammation.

[0435] In the implementation plan, the disorder is selected from liver disorder and kidney disorder.

[0436] In the implementation plan, liver disorders are selected from: liver fibrosis occurring in end-stage liver disease; cirrhosis; liver failure due to toxicity; non-alcoholic steatosis or NASH; and alcoholic steatosis.

[0437] In the implementation plan, kidney disorder is defined as kidney fibrosis.

[0438] In the implementation plan, renal fibrosis occurs due to acute kidney injury.

[0439] In the implementation plan, kidney disorder is defined as chronic kidney disorder.

[0440] In the implementation plan, kidney dysfunction refers to diabetic nephropathy.

[0441] In another aspect, this disclosure provides a method for treating an age-related disorder or disease in a subject with a corresponding need, the method comprising administering to the subject a therapeutically effective amount of a compound of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic) or a pharmaceutically acceptable salt thereof, a pharmaceutical composition comprising a compound of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a compound of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic) or a pharmaceutically acceptable salt thereof, wherein the disorder or disease is selected from: sarcopenia, skin atrophy, cherry angioma, seborrheic keratosis, cerebral atrophy, atherosclerosis, arteriosclerosis, pulmonary atrophy. Emphysema, osteoporosis, osteoarthritis, hypertension, erectile dysfunction, cataracts, macular degeneration, glaucoma, stroke, cerebrovascular disease (stroke), chronic kidney disease, diabetic nephropathy, impaired liver function, liver fibrosis, autoimmune hepatitis, endometrial hyperplasia, metabolic dysfunction, renal vascular disease, hearing loss, activity deficits, cognitive decline, tendon stiffness, cardiac dysfunction such as cardiomegaly and / or systolic and / or diastolic dysfunction and / or hypertension, cardiac dysfunction leading to decreased ejection fraction, immunosenescence, Parkinson's disease, Alzheimer's disease, cancer, immunosenescence leading to cancer due to decreased immune surveillance, infections due to decreased immune function, chronic obstructive pulmonary disease (COPD), obesity, loss of taste, loss of smell, arthritis, and type II diabetes (including complications arising from diabetes such as kidney failure, blindness, and neurological disorders).

[0442] In another aspect, this disclosure provides a method of treating a subject with cancer, the method comprising administering to the subject a therapeutically effective amount of a compound of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic) or a pharmaceutically acceptable salt thereof, a pharmaceutical composition comprising a compound of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a compound of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic) or a pharmaceutically acceptable salt thereof.

[0443] In the implementation plan, the method also includes a PD-1 / PDL-1 inhibitor.

[0444] In the implementation plan, the cancers are selected from kidney cancer, renal cell carcinoma, colorectal cancer, uterine sarcoma, endometrial cancer, breast cancer, ovarian cancer, cervical cancer, gastric cancer, fibrosarcoma, pancreatic cancer, liver cancer, melanoma, leukemia, multiple myeloma, nasopharyngeal carcinoma, prostate cancer, lung cancer, glioblastoma, bladder cancer, mesothelioma, head cancer, rhabdomyosarcoma, sarcoma, lymphoma, and cervical cancer.

[0445] In another aspect, this disclosure provides compounds of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic) or pharmaceutically acceptable salts thereof, pharmaceutical compositions comprising compounds of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic) or pharmaceutically acceptable salts thereof, or pharmaceutical compositions comprising compounds of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic) or pharmaceutically acceptable salts thereof, for use as a medicament.

[0446] In another aspect, this disclosure provides compounds of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic), or pharmaceutically acceptable salts thereof, pharmaceutical compositions comprising compounds of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic), or pharmaceutical compositions comprising compounds of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic), or pharmaceutical compositions comprising pharmaceutically acceptable salts thereof, for use in the prevention or treatment of disorders or diseases mediated by the mTOR pathway.

[0447] In another aspect, this disclosure provides compounds of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic), or pharmaceutically acceptable salts thereof, pharmaceutical compositions comprising compounds of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(Ic), or pharmaceutically acceptable salts thereof, or pharmaceutical compositions comprising compounds of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic), or pharmaceutically acceptable salts thereof, for use in the prevention or treatment of disorders or diseases selected from:

[0448] • Acute or chronic organ or tissue graft rejection;

[0449] • Transplant vascular disease;

[0450] • Smooth muscle cell proliferation and migration lead to thickening of the vascular intima, vascular occlusion, obstructive coronary atherosclerosis, and restenosis;

[0451] • Autoimmune diseases and inflammatory conditions;

[0452] ·asthma;

[0453] Multidrug resistance (MDR);

[0454] Fungal infection;

[0455] Inflammation;

[0456] ·Infect;

[0457] Age-related diseases;

[0458] Neurodegenerative diseases;

[0459] • Proliferative disorders, especially cancer;

[0460] • Seizures and seizure-related disorders; and

[0461] • Mitochondrial myopathy and mitochondrial stress.

[0462] In another aspect, this disclosure provides compounds of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic), or pharmaceutically acceptable salts thereof, pharmaceutical compositions comprising compounds of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic), or pharmaceutically acceptable salts thereof, or pharmaceutical compositions comprising compounds of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic), or pharmaceutically acceptable salts thereof, for use in the prevention or treatment of disorders or diseases including processes of fibrosis and / or inflammation.

[0463] In the implementation plan, the disorder is selected from liver disorder and kidney disorder.

[0464] In the implementation plan, liver disorders are selected from: liver fibrosis occurring in end-stage liver disease; cirrhosis; liver failure due to toxicity; non-alcoholic steatosis or NASH; and alcoholic steatosis.

[0465] In the implementation plan, renal disorder is renal fibrosis that occurs due to acute kidney injury.

[0466] In the implementation plan, kidney disorder is defined as chronic kidney disorder.

[0467] In the implementation plan, kidney dysfunction refers to diabetic nephropathy.

[0468] In another aspect, this disclosure provides compounds of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic), or pharmaceutically acceptable salts thereof, pharmaceutical compositions comprising compounds of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic), or pharmaceutically acceptable salts thereof, or pharmaceutical compositions comprising compounds of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic), or pharmaceutically acceptable salts thereof, for use in the treatment of age-related disorders or diseases selected from: sarcopenia, skin atrophy, cherry angioma, seborrheic keratosis, cerebral atrophy (also known as dementia), atherosclerosis, arteriosclerosis, emphysema, osteoporosis, osteoarthritis, and high blood pressure. Pressure, erectile dysfunction, cataracts, macular degeneration, glaucoma, stroke, cerebrovascular disease (stroke), chronic kidney disease, diabetic nephropathy, impaired liver function, liver fibrosis, autoimmune hepatitis, endometrial hyperplasia, metabolic dysfunction, renal vascular disease, hearing loss, activity deficits (e.g., weakness), cognitive decline, tendon stiffness, cardiac dysfunction such as cardiac hypertrophy and / or systolic and / or diastolic dysfunction and / or hypertension, cardiac dysfunction leading to decreased ejection fraction, immunosenescence, Parkinson's disease, Alzheimer's disease, cancer, immunosenescence leading to cancer due to decreased immune surveillance, infections due to decreased immune function, chronic obstructive pulmonary disease (COPD), obesity, loss of taste, loss of smell, arthritis, and type II diabetes (including complications arising from diabetes such as kidney failure, blindness, and neurological disorders).

[0469] In another aspect, this disclosure provides compounds of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic), or pharmaceutically acceptable salts thereof, pharmaceutical compositions comprising compounds of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic), or pharmaceutically acceptable salts thereof, or pharmaceutical compositions comprising compounds of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic), or pharmaceutically acceptable salts thereof, for use in the treatment of cancer.

[0470] In another aspect, this disclosure provides compounds of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic), or pharmaceutically acceptable salts thereof, pharmaceutical compositions comprising compounds of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic), or pharmaceutically acceptable salts thereof, or compositions comprising formula (I), (P-1), (P-2), (Ia)-(Ic). A combination of (P-Ia)-(P-Ic) compounds or pharmaceutically acceptable salts thereof, for use in the treatment of renal cell carcinoma, renal cell carcinoma, colorectal cancer, uterine sarcoma, endometrial cancer, breast cancer, ovarian cancer, cervical cancer, gastric cancer, fibrosarcoma, pancreatic cancer, liver cancer, melanoma, leukemia, multiple myeloma, nasopharyngeal carcinoma, prostate cancer, lung cancer, glioblastoma, bladder cancer, mesothelioma, head cancer, rhabdomyosarcoma, sarcoma, lymphoma, or cervical cancer.

[0471] In another aspect, this disclosure provides the use of compounds of formula (I), (P-1), (P-2), (Ia)-(Ic) or (P-Ia)-(P-Ic) or pharmaceutically acceptable salts thereof for the manufacture of medicaments.

[0472] In another aspect, this disclosure provides the use of compounds of formula (I), (P-1), (P-2), (Ia)-(Ic) or (P-Ia)-(P-Ic) or pharmaceutically acceptable salts thereof for the manufacture of medicaments for the treatment of disorders or diseases mediated by the mTOR pathway.

[0473] In another aspect, this disclosure provides the use of compounds of formula (I), (P-1), (P-2), (Ia)-(Ic) or (P-Ia)-(P-Ic) or pharmaceutically acceptable salts thereof for the manufacture of a medicament for the treatment of disorders or diseases selected from:

[0474] • Acute or chronic organ or tissue graft rejection;

[0475] • Transplant vascular disease;

[0476] • Smooth muscle cell proliferation and migration lead to thickening of the vascular intima, vascular occlusion, obstructive coronary atherosclerosis, and restenosis;

[0477] • Autoimmune diseases and inflammatory conditions;

[0478] ·asthma;

[0479] Multidrug resistance (MDR);

[0480] Fungal infection;

[0481] Inflammation;

[0482] ·Infect;

[0483] Age-related diseases;

[0484] Neurodegenerative diseases;

[0485] • Proliferative disorders, such as cancer;

[0486] • Seizures and seizure-related disorders; and

[0487] • Mitochondrial myopathy and mitochondrial stress.

[0488] In another aspect, this disclosure provides the use of compounds of formula (I), (P-1), (P-2), (Ia)-(Ic) or (P-Ia)-(P-Ic) or pharmaceutically acceptable salts thereof for the manufacture of medicaments for the treatment of disorders or diseases including processes of fibrosis or inflammation.

[0489] In the implementation plan, the disorder is selected from liver disorder and kidney disorder.

[0490] In the implementation plan, liver disorders are selected from: liver fibrosis occurring in end-stage liver disease; cirrhosis; liver failure due to toxicity; non-alcoholic steatosis or NASH; and alcoholic steatosis.

[0491] In the implementation plan, renal disorder is renal fibrosis that occurs due to acute kidney injury.

[0492] In the implementation plan, kidney disorder is defined as chronic kidney disorder.

[0493] In the implementation plan, kidney dysfunction refers to diabetic nephropathy.

[0494] In another aspect, this disclosure provides the use of compounds of formula (I), (P-1), (P-2), (Ia)-(Ic), or (P-Ia)-(P-Ic) or pharmaceutically acceptable salts thereof for the manufacture of medicaments for the prevention or treatment of age-related disorders or diseases selected from: sarcopenia, skin atrophy, cherry angioma, seborrheic keratosis, brain atrophy (also known as dementia), atherosclerosis, arteriosclerosis, emphysema, osteoporosis, osteoarthritis, hypertension, erectile dysfunction, cataracts, macular degeneration, glaucoma, stroke, cerebrovascular disease (stroke), chronic kidney disease, diabetic nephropathy, impaired liver function, etc. Liver fibrosis, autoimmune hepatitis, endometrial hyperplasia, metabolic dysfunction, renal vascular disease, hearing loss, activity deficits, cognitive decline, tendon stiffness, cardiac dysfunction such as cardiomegaly and / or systolic and / or diastolic dysfunction and / or hypertension, cardiac dysfunction leading to decreased ejection fraction, immunosenescence, Parkinson's disease, Alzheimer's disease, cancer, immunosenescence leading to cancer due to decreased immune surveillance, infections due to decreased immune function, chronic obstructive pulmonary disease (COPD), obesity, anosmia, anosmia, arthritis, and type II diabetes (including complications arising from diabetes such as kidney failure, blindness, and neurological disorders).

[0495] In another aspect, this disclosure provides the use of compounds of formula (I), (P-1), (P-2), (Ia)-(Ic) or (P-Ia)-(P-Ic) or pharmaceutically acceptable salts thereof for the manufacture of medicaments for the prevention or treatment of cancer.

[0496] In another aspect, this disclosure provides the use of compounds of formula (I), (P-1), (P-2), (Ia)-(Ic) or (P-Ia)-(P-Ic) or pharmaceutically acceptable salts thereof for the manufacture of medicaments for the treatment of renal cell carcinoma, colorectal cancer, uterine sarcoma, endometrial cancer, breast cancer, ovarian cancer, cervical cancer, gastric cancer, fibrosarcoma, pancreatic cancer, liver cancer, melanoma, leukemia, multiple myeloma, nasopharyngeal carcinoma, prostate cancer, lung cancer, glioblastoma, bladder cancer, mesothelioma, head cancer, rhabdomyosarcoma, sarcoma, lymphoma, or cervical cancer.

[0497] This document sets forth details of one or more embodiments of the present disclosure. Other features, objects, and advantages of the present disclosure will become apparent from the embodiments, figures / drawings, and claims. Example

[0498] Some embodiments of this disclosure are illustrated by the following non-limiting examples. As used herein, the symbols and conventions used in these processes, methods, and examples, whether or not specific abbreviations are specifically defined, are consistent with those used in contemporary scientific literature, such as the Journal of the American Chemical Society or the Journal of Biochemistry. Specifically, but not in a limiting sense, the following abbreviations may be used in the examples and throughout the specification.

[0499]

[0500]

[0501] Example 1. Synthesis of 3,2-deoxyrapamycin:

[0502]

[0503] Step 1: 2,6-Dimethylpyridine (15.29 mL, 131.26 mmol) and TESOTf (14.84 mL, 65.63 mmol) were added to a solution of rapamycin (20 g, 21.88 mmol) in DCM (280 mL) at -40 °C and stirred for 1.5 h. The resulting mixture was diluted with Et₂O and quenched with saturated NaHCO₃ aqueous solution. The organic phase was separated and washed with saturated NaHCO₃ aqueous solution, saturated CuSO₄ aqueous solution, and saturated NH₄Cl aqueous solution. The combined aqueous phase was re-extracted with Et₂O. The combined organic phase was washed with saturated CuSO₄ aqueous solution, dried over MgSO₄, filtered, and concentrated under vacuum. The residue was dissolved in DCM and subjected to an ISCO silica column, followed by elution with a gradient of 0% to 25% EtOAc / hexane to obtain 95% of the intermediate 1((3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-27-hydroxy-10,21-dimethoxy-3-((R)-1-((1S,3R,4R)-3-methoxy-4-(( Triethylsilyl)oxy)cyclohexyl)propyl-2-yl)-6,8,12,14,20,26-hexamethyl-9-((triethylsilyl)oxy)-9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-hexadecylhydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclotriacontria-1,5,11,28,29(4H,6H,31H)-pentaone). 1The H-NMR data matched the reference (i.e., Tetrahedron Letters, 1994, 35(41), 7557-7560).

[0504] Step 2: Intermediate product 2((3S,5R,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-5,27-dihydroxy-10,21-dimethoxy-3-((R)-1-((1S,3R,4R)-3-methoxy-4-((triethylsilyl)oxy)cyclohexyl)propyl-2-yl)-6,8,12 ,14,20,26-hexamethyl-9-((triethylsilyl)oxy)-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-octadecylhydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohexadecene-1,11,28,29(4H,31H)-tetraone) was prepared according to the procedure in patent WO 96 / 41807 and matched 1 H-NMR data.

[0505] Step 3: Intermediate product 3((3S,5R,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-27-hydroxy-10,21-dimethoxy-3-((R)-1-((1S,3R,4R)-3-methoxy-4-((triethylsilyl)oxy)cyclohexyl)propyl-2-yl)-6,8,12,14,20,26- Hexamethyl-1,11,28,29-tetraoxo-9-((triethylsilyl)oxy)-1,4,5,6,9,10,11,12,13,14,21,22,23,24,25,26,27,28,29,31,32,33,34,34a-tetrahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohexane-5-ylmethanesulfonate) was prepared according to the procedure in patent WO 96 / 41807 and matched 1 H-NMR data.

[0506] Step 4: Intermediate product 4((3S,5S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-27-hydroxy-5-iodo-10,21-dimethoxy-3-((R)-1-((1S,3R,4R)-3-methoxy-4-((triethylsilyl)oxy)cyclohexyl)propyl-2-yl)-6,8,1 2,14,20,26-hexamethyl-9-((triethylsilyl)oxy)-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-octadecylhydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohexadecene-1,11,28,29(4H,31H)-tetraone) was prepared according to the procedure in patent WO 96 / 41807 and matched 1 H-NMR data.

[0507] Step 5: Intermediate product 5((3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-27-hydroxy-10,21-dimethoxy-3-((R)-1-((1S,3R,4R)-3-methoxy-4-((triethylsilyl)oxy)cyclohexyl)propyl-2-yl)-6,8,12,1 4,20,26-Hexamethyl-9-((triethylsilyl)oxy)-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-octadecylhydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohexadecene-1,11,28,29(4H,31H)-tetraone was prepared according to the procedure in patent WO 96 / 41807 and matched 1 H-NMR data.

[0508] Step 6: 32-Deoxyrapamycin was prepared according to the procedure in patent WO 96 / 41807. Purification was performed by eluting with an ISO silica column using a gradient of 40% to 60% EtOAc / hexane to provide 95% of the product as a white solid. 1 The H-NMR data and quality match those in the references.

[0509] Example 2. Synthesis of a representative 32-deoxyrapamycin analog

[0510] General conditions A1: ZnCl2 (3 equivalents, 1.0 M Et2O solution) was added to a solution of 3,2-deoxyrapamycin (30.0 mg, 0.0333 mmol) and an appropriate amine nucleophile (5 equivalents) in DCM (0.67 mL) at 0 °C. The reaction was heated to rt and stirred until the 3,2-deoxyrapamycin was consumed. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The organic phase was separated and the aqueous phase was extracted three times with EtOAc. The combined organic phases were dried over Na2SO4, filtered, and concentrated under vacuum. The residue was dissolved in DMF and subjected to an ISCO C18 RP column and eluted with a gradient of 60% to 85% ACN / water (+0.05% AcOH as a modifier) ​​to obtain the product.

[0511] General Condition B1: Add pTSA·H₂O (5 equivalents) to a solution of 32-deoxyrapamycin (30.0 mg, 0.0333 mmol) and a suitable amine nucleophile (20 equivalents) in DCM (0.67 mL) at rt. Stir the reaction at rt until the 32-deoxyrapamycin is consumed. Dilute the resulting mixture with EtOAc and quench it with a saturated aqueous solution of NaHCO₃. Separate the organic phase and extract the aqueous phase three times with EtOAc. Dry the combined organic phases with Na₂SO₄, filter, and concentrate under vacuum. Dissolve the residue in DMF and precipitate it on an ISCO C18 RP column, eluting with a gradient of 60% to 85% ACN / water (+0.05% AcOH as a modifier) ​​to obtain the product.

[0512] Compound 3:

[0513]

[0514] Under general conditions A1, the reaction was carried out. The procedure was modified using 20 equivalents of triazole and 15 equivalents of ZnCl2. An additional 1.0 mL of ACN was added as a co-solvent. The reaction was stirred at 39°C for 5 days to complete. Compound 3 was isolated as a white solid at 6.1%. Calculated mass: 936.58; Found mass: 935.93 (MH). - .

[0515] Compound 4:

[0516]

[0517] General conditions A1 were applied. The program was modified with 10 equivalents of pyrazole and 10 equivalents of ZnCl2. The reaction was stirred at rt for 2 days to complete. Compound 4 was isolated as a white solid at 13%. Calculated mass: 935.59 g / mL; Found mass: 934.93 g / mL (MH). - .

[0518] Compound 5:

[0519]

[0520] Under general conditions A1, the reaction was stirred at rt for 2 hours to complete. Compound 5 was separated as a white solid at 32%. Calculated mass: 1002.59 g / mL; Found mass: 1001.99 g / mL (MH). - .

[0521] Compound 6:

[0522]

[0523] Under general conditions A1, the reaction was modified using 30 equivalents of triazole and 10 equivalents of ZnCl2. An additional 2.0 mL of THF was added as a co-solvent. The reaction was stirred at 39°C for 2 days to complete. Compound 6 was isolated as a white solid at 10%. Calculated mass: 936.58; Found mass: 938.06 (M+H). + .

[0524] Compound 8:

[0525]

[0526] Under general conditions A1, the program was modified using 20 equivalents of tetrazolium and 15 equivalents of ZnCl2. The reaction was stirred at rt for 1 h to complete. Compound 8 was separated as a white solid at 11%. Calculated mass: 937.58; Found mass: 939.05 (M+H). + .

[0527] Compound 9:

[0528]

[0529] Under general conditions A1, the reaction was stirred at rt for 2 hours to complete. Compound 9 was separated as a white solid at 25%. Calculated mass 1004.60, Found mass 1004.04 (MH). - .

[0530] Compound 10:

[0531]

[0532] Under general conditions A1, the reaction was stirred at rt for 2 hours to complete. Compound 10 was separated as a white solid at 18%. Calculated mass: 990.59 g / mL; Measured mass: 989.68 g / mL (MH). - .

[0533] Compound 11:

[0534]

[0535] Under general conditions A1, an additional 0.6 mL of ACN was added as a co-solvent. The reaction was stirred at rt for 6 h to complete. Compound 11 was separated as a white solid at 9.5%. Calculated mass 1006.58, Found mass 1006.06 (MH). - .

[0536] Compound 12:

[0537]

[0538] Under general conditions A1, add an additional 0.6 mL of ACN as a co-solvent. Stir the reaction at rt for 2 h to complete. Compound 12 was separated as a white solid at 8.3%. Calculated mass 1005.63, Found mass 1006.73 (M+H) + .

[0539] Compound 13:

[0540]

[0541] Under general conditions A1, an additional 0.6 mL of ACN was added as a co-solvent. The reaction was stirred at rt for 16 h to complete. Compound 14 was separated as a white solid at 22%. Calculated mass 991.61, Found mass 991.01 (MH). - .

[0542] Compound 14:

[0543]

[0544] General conditions A1 were applied. The procedure was modified with 10 equivalents of sulfonamide and 6 equivalents of ZnCl2. The reaction was stirred at rt for 5 days to complete. Compound 14 was isolated as a yellow solid at 11%. Calculated mass 951.58, Found mass 950.89 (MH). - .

[0545] Compound 15:

[0546]

[0547] Under general conditions A1, the reaction was stirred at rt for 16 h to complete. Compound 15 was separated as a white solid at 5.7%. Calculated mass: 978.63; Found mass: 979.92 (M+H). + .

[0548] The following compounds are prepared under general conditions A2, B2, C2, D2 or E2.

[0549] General conditions A2: Add ZnCl2 (10 equivalents, 1.0 M Et2O solution) to a solution of 3,2-deoxyrapamycin (30.0 mg, 0.0333 mmol) and a suitable amine nucleophile (10 equivalents) in DCM (0.67 mL). Stir the reaction until the 3,2-deoxyrapamycin is consumed. Dilute the resulting mixture with EtOAc and quench it with a saturated aqueous solution of NaHCO3. Separate the organic phase and extract the aqueous phase three times with EtOAc. Dry the combined organic phases with Na2SO4, filter, and concentrate under vacuum. Dissolve the residue in DMF and precipitate it on an ISCO C18 RP column, eluting with a gradient of (30–60)% to (80–90)% ACN / water (+0.05% TFA as a modifier) ​​to obtain the product.

[0550] General Condition B2: Add pTSA·H₂O (5 equivalents) to a solution of 32-deoxyrapamycin (30.0 mg, 0.0333 mmol) and a suitable amine nucleophile (20 equivalents) in DCM (0.67 mL). Stir the reaction at rt until the 32-deoxyrapamycin is consumed. Dilute the resulting mixture with EtOAc and quench it with a saturated aqueous solution of NaHCO₃. Separate the organic phase and extract the aqueous phase three times with EtOAc. Dry the combined organic phases with Na₂SO₄, filter, and concentrate under vacuum. Dissolve the residue in DMF and precipitate it on an ISCO C18 RP column, eluting with a gradient of 60% to 85% ACN / water (+0.05% AcOH as a modifier) ​​to obtain the product.

[0551] General conditions C2: Add BF3·Et2O (15 equivalents) to a solution of 32-deoxyrapamycin (30.0 mg, 0.0333 mmol) and a suitable amine nucleophile (10 equivalents) in THF (0.67 mL) at rt. Stir the reaction at rt until the 32-deoxyrapamycin is consumed. Dilute the resulting mixture with EtOAc and quench it with a saturated aqueous solution of NaHCO3. Separate the organic phase and extract the aqueous phase three times with EtOAc. Dry the combined organic phases with Na2SO4, filter, and concentrate under vacuum. Dissolve the residue in DMF and precipitate it on an ISCO C18 RP column, eluting with a gradient of (30–60)% to (80–90)% ACN / water (+0.05% AcOH as a modifier) ​​to obtain the product.

[0552] General conditions D2: Zn(OTf)₂ (6 equivalents) was added to a solution of 32-deoxyrapamycin (60.0 mg, 0.0667 mmol) and a suitable amine nucleophile (10 equivalents) in DCM / ACN (1:1) (2.7 mL) at rt. The reaction was stirred at rt until all 32-deoxyrapamycin was consumed. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO₃. The organic phase was separated and the aqueous phase was extracted three times with EtOAc. The combined organic phases were dried over Na₂SO₄, filtered, and concentrated under vacuum. The residue was dissolved in DMF and subjected to an ISCO C18 RP column and eluted with a gradient of 50% to 95% ACN / water (+0.05% AcOH as a modifier) ​​to obtain the product.

[0553] General conditions E2: Add TFA (20 equivalents) to a solution of 32-deoxyrapamycin (60.0 mg, 0.0667 mmol) and a suitable amine nucleophile (10 equivalents) in DCM (3 mL) at -40 °C and allow the reaction to rise to rt. Stir the reaction at rt until the 32-deoxyrapamycin is consumed. Dilute the resulting mixture with EtOAc and quench it with a saturated aqueous solution of NaHCO3. Separate the organic phase and extract the aqueous phase three times with EtOAc. Dry the combined organic phases with Na2SO4, filter, and concentrate under vacuum. Dissolve the residue in DMF and precipitate it on an ISCO C18 RP column, eluting with a gradient of 30% to 80% ACN / water (+0.05% AcOH as a modifier) ​​to obtain the product.

[0554] Compound 16:

[0555]

[0556] Under general conditions A2, ZnCl2 (10 equivalents, 1.0 M Et2O solution) was added to a solution of 3,2-deoxyrapamycin (30.0 mg, 0.0333 mmol) and sulfonamide nucleophile (10 equivalents) in DCM (0.67 mL). The reaction was stirred until the 3,2-deoxyrapamycin was consumed. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The organic phase was separated and the aqueous phase was extracted three times with EtOAc. The combined organic phases were dried over Na2SO4, filtered, and concentrated under vacuum. The residue was dissolved in DMF and subjected to an ISCO C18 RP column and eluted with a gradient of (30–60)% to (80–90)% ACN / water (+0.05% TFA as a modifier) ​​to obtain the product. The reaction was stirred for 1 h to complete. Compound 16 was isolated and further purified by a 150 × 30 Phenomenex Gemini reversed-phase column with elution from 40% to 80% to give a product as a white solid with a purity of 15.5% and a mixture in a yield of 30.2%. Calculated mass 1023.56, Found 1024.56 (MH). - .

[0557] Compound 17:

[0558]

[0559] Under general conditions A2, ZnCl2 (10 equivalents, 1.0 M Et2O solution) was added to a solution of 32-deoxyrapamycin (30.0 mg, 0.0333 mmol) and sulfonamide nucleophile (10 equivalents) in DCM (0.67 mL). The reaction was stirred until the 32-deoxyrapamycin was consumed. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The organic phase was separated and the aqueous phase was extracted three times with EtOAc. The combined organic phases were dried over Na2SO4, filtered, and concentrated under vacuum. The residue was dissolved in DMF and subjected to an ISCO C18 RP column and eluted with a gradient of (30–60)% to (80–90)% ACN / water (+0.05% TFA as a modifier) ​​to obtain the product. The reaction was stirred for 4 days to complete. Compound 17 was isolated and further purified by a 150 × 30 Phenomenex Gemini reversed-phase column, eluted with 35% to 75% to give a product as a white solid with a purity of 1.4% and a mixture in 5% yield. Calculated mass 1026.57, Found 1027.30 (M+H) + .

[0560] Compound 18:

[0561]

[0562] Under general conditions C2, add BF3·Et2O (15 equivalents) to a solution of 32-deoxyrapamycin (30.0 mg, 0.0333 mmol) and sulfonamide nucleophile (10 equivalents) in THF (0.67 mL). Stir the reaction at rt until the 32-deoxyrapamycin is consumed. Dilute the resulting mixture with EtOAc and quench it with a saturated aqueous solution of NaHCO3. Separate the organic phase and extract the aqueous phase three times with EtOAc. Dry the combined organic phases with Na2SO4, filter, and concentrate under vacuum. Dissolve the residue in DMF and precipitate it on an ISCO C18 RP column, eluting with a gradient of (30–60)% to (80–90)% ACN / water (+0.05% AcOH as a modifier) ​​to obtain the product. Stir the reaction for 5 hours to complete. Compound 18 was isolated and further purified by a 150 × 30 Phenomenex Gemini reversed-phase column with elution from 30% to 80%, yielding the product as a pale yellow solid in 12.4% yield. Calculated mass 1024.56, Found 1025.13 (MH). - .

[0563] Compound 19:

[0564]

[0565] Under general conditions C2, add BF3·Et2O (15 equivalents) to a solution of 32-deoxyrapamycin (30.0 mg, 0.0333 mmol) and sulfonamide nucleophile (10 equivalents) in THF (0.67 mL). Stir the reaction at rt until the 32-deoxyrapamycin is consumed. Dilute the resulting mixture with EtOAc and quench it with a saturated aqueous solution of NaHCO3. Separate the organic phase and extract the aqueous phase three times with EtOAc. Dry the combined organic phases with Na2SO4, filter, and concentrate under vacuum. Dissolve the residue in DMF and precipitate it on an ISCO C18 RP column, eluting with a gradient of (30–60)% to (80–90)% ACN / water (+0.05% AcOH as a modifier) ​​to obtain the product. Stir the reaction for 5 hours to complete. Compound 19 was isolated and further purified by a 150 × 30 Phenomenex Gemini reversed-phase column with elution from 30% to 80%, yielding the product as a pale yellow solid in 7% yield. Calculated mass 1024.56, Found 1026.46 (MH). - .

[0566] Compound 20:

[0567]

[0568] Under general conditions A, ZnCl₂ (15 equivalents) and ACN (0.67 mL) were added to the reaction, and the reaction was stirred for 3 days to complete. Compound 20 was isolated and further purified by elution using a 150 × 30 Phenomenex Gemini reversed-phase column with 40% to 80% elution, yielding the product as a yellow solid in 7.3% yield. Calculated mass 962.61, Found mass 963.31 (M+H) + .

[0569] Compound 21:

[0570]

[0571] Under general conditions C2, add BF3·Et2O (15 equivalents) to a solution of 32-deoxyrapamycin (30.0 mg, 0.0333 mmol) and an amine nucleophile (10 equivalents) in THF (0.67 mL). Stir the reaction at rt until the 32-deoxyrapamycin is consumed. Dilute the resulting mixture with EtOAc and quench it with a saturated aqueous solution of NaHCO3. Separate the organic phase and extract the aqueous phase three times with EtOAc. Dry the combined organic phases with Na2SO4, filter, and concentrate under vacuum. Dissolve the residue in DMF and precipitate it on an ISCO C18 RP column, eluting with a gradient of (30–60)% to (80–90)% ACN / water (+0.05% AcOH as a modifier) ​​to obtain the product. Stir the reaction for 3 hours to complete. Compound 21 was isolated and further purified by a 150 × 30 Phenomenex Gemini reversed-phase column with elution from 30% to 80% to give a product as a white solid in 9% yield and a mixture in 10.2% yield. Calculated mass 962.61, Found mass 963.31 (M+H) + .

[0572] Compound 22:

[0573]

[0574] Under general conditions C2, BF3·Et2O (15 equivalents) was added to a solution of 32-deoxyrapamycin (30.0 mg, 0.0333 mmol) and the corresponding amine nucleophile (10 equivalents) in THF (0.67 mL) at rt. The reaction was stirred at rt until the 32-deoxyrapamycin was consumed. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The organic phase was separated and the aqueous phase was extracted three times with EtOAc. The combined organic phases were dried over Na2SO4, filtered, and concentrated under vacuum. The residue was dissolved in DMF and subjected to an ISCO C18 RP column and eluted with a gradient of (30–60)% to (80–90)% ACN / water (+0.05% AcOH as a modifier) ​​to obtain the product. The reaction was stirred for 3 hours to complete. Compound 22 was isolated and further purified by a 150 × 30 Phenomenex Gemini reversed-phase column with elution from 30% to 80% to give a product as a white solid in yield of 1.54% and a mixture in yield of 2.89%. Calculated mass 962.61, Found 963.28 (M+H) + .

[0575] Compound 23:

[0576]

[0577] Under general conditions E2, TFA (20 equivalents) was added to a solution of 32-deoxyrapamycin (60.0 mg, 0.0667 mmol) and an amine nucleophile (10 equivalents) in DCM (3 mL) at -40 °C, allowing to reach rt. The reaction was stirred at rt until 32-deoxyrapamycin was consumed. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The organic phase was separated and the aqueous phase was extracted three times with EtOAc. The combined organic phases were dried over Na2SO4, filtered, and concentrated under vacuum. The residue was dissolved in DMF and subjected to an ISCO C18 RP column, and eluted with a gradient of 30% to 80% ACN / water (+0.05% AcOH as a modifier) ​​to obtain the product. The reaction was stirred for 19 hours to complete. Compound 23 was isolated and further purified by a 150 × 30 Phenomenex Gemini reversed-phase column with elution from 30% to 80% to give a product in 7.6% yield as a brown solid and a mixture in 5.6% yield. Calculated mass 965.62, Found 966.35 (M+H) + .

[0578] Compound 24:

[0579]

[0580] Under general conditions E2, TFA (20 equivalents) was added to a solution of 32-deoxyrapamycin (60.0 mg, 0.0667 mmol) and an amine nucleophile (10 equivalents) in DCM (3 mL) at -40 °C, allowing to reach rt. The reaction was stirred at rt until 32-deoxyrapamycin was consumed. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The organic phase was separated and the aqueous phase was extracted three times with EtOAc. The combined organic phases were dried over Na2SO4, filtered, and concentrated under vacuum. The residue was dissolved in DMF and subjected to an ISCO C18 RP column, and eluted with a gradient of 30% to 80% ACN / water (+0.05% AcOH as a modifier) ​​to obtain the product. The reaction was stirred for 2 hours to complete. Compound 24 was isolated and further purified by a 150 × 30 Phenomenex Gemini reversed-phase column with elution from 30% to 80% to give a product in 14.9% as a brown solid and a mixture in 21.6% yield. Calculated mass 950.57, Found 951.08 (MH). - .

[0581] Compound 26(3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methoxy-6,8,12,14,20,26-hexamethyl-2 1-(phenylamino)-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-octadecylhydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohexadecene-1,11,28,29(4H,31H)-tetraone; 16-aniline-32-deoxyrapamycin:

[0582]

[0583] Aniline (0.080 mL, 0.888 mmol) and TFA (0.136 mL, 1.776 mmol) were added to a stirred solution of 32-deoxyrapamycin (80 mg, 0.088 mmol) in DCM (2 mL) at -20 °C and stirred for 2 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCO C18 column, eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 7.3 mg (9%) of compound 26 as a white solid. 56 H 84 N2O 11 The calculated MS(ESI) value for +H is 960.61, and the measured value is 960.08.

[0584] Compound 27(3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methoxy-6,8,12,14,20,26-hexamethyl-21-((5-methylisoxazole- 3-yl)amino)-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-octadecylhydro-3H-23,27-epoxypyridino[2,1-c][1]oxa[4]azacyclohexadecene-1,11,28,29(4H,31H)-tetraone; 16-(3-amino-5-methylisoxazole)-32-deoxyrapamycin:

[0585]

[0586] 3-Amino-5-methylisoxazole (87 mg, 0.888 mmol) and TFA (0.136 mL, 1.779 mmol) were added to a stirred solution of 32-deoxyrapamycin (80 mg, 0.088 mmol) in DCM (2 mL) at -20 °C and stirred for 2 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCO C18 column, eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 5.8 mg (7%) of the compound as a white solid. 54H 83 N3O 12 The calculated MS(ESI) value for +H is 965.61, and the measured value is 965.10.

[0587] Compound 28(3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methoxy-6,8,12,14,20,26-hexamethyl-21-((6-methylpyrazine- 2-yl)amino)-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-octadecylhydro-3H-23,27-epoxypyridino[2,1-c][1]oxa[4]azacyclohexadecene-1,11,28,29(4H,31H)-tetraone; 16-(2-amino-6-methylpyrazine)-32-deoxyrapamycin:

[0588]

[0589] 2-Amino-6-methylpyrazine (61 mg, 0.555 mmol) and TFA (0.085 mL, 1.112 mmol) were added to a stirred solution of 32-deoxyrapamycin (50 mg, 0.055 mmol) in DCM (2 mL) at -20 °C and stirred for 2 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCO C18 column, eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 7.2 mg (13%) of a white solid compound 28. 55 H 84 N4O 11 The calculated MS(ESI) value for +H is 976.61, and the measured value is 976.31.

[0590] Compound 29, (3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methoxy-6,8,12,14,20,26-hexamethyl -21-((1-methyl-1H-pyrazol-5-yl)amino)-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-octadecylhydro-3H-23,27-epoxypyridino[2,1-c][1]oxa[4]azacyclohexadecene-1,11,28,29(4H,31H)-tetraone

[0591]

[0592] 1,2,5-oxadiazol-3-amine (66 mg, 0.666 mmol) and TFA (0.102 mL, 1.322 mmol) were added to a stirred solution of 32-deoxyrapamycin (60 mg, 0.066 mmol) in DCM (2 mL) at -20 °C and stirred for 2 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCO C18 column, eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 1.3 mg (2%) of white solid. 52 H 80 N4O 12 The calculated MS(ESI-) value for -H is 964.61, and the measured value is 964.11.

[0593] Compound 30(3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-21-((1,2,4-oxadiazol-3-yl)amino)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methyl oxy-6,8,12,14,20,26-hexamethyl-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-octadecylhydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohexadecene-1,11,28,29(4H,31H)-tetraone;

[0594] (16-(1,2,4-oxadiazol-3-amine)-32-deoxyrapamycin:

[0595]

[0596] 1,2,4-oxadiazol-3-amine (55 mg, 0.555 mmol) and TFA (0.085 mL, 1.112 mmol) were added to a stirred solution of 32-deoxyrapamycin (50 mg, 0.055 mmol) in DCM (2 mL) at -20 °C and stirred for 2 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCO C18 column and eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 5.3 mg (10%) of a white solid compound 30. 52 H 80 N4O 12 The calculated MS(ESI) value for +H is 952.58, and the measured value is 952.17.

[0597] The synthesis of another compound is described below.

[0598] Compound 31:

[0599]

[0600] Dimethylphosphonochloride (12.5 mg, 0.111 mmol) was added to a solution of 32-deoxyrapamycin (20.0 mg, 0.0222 mmol) and 2,6-di-tert-butyl-4-methylpyridine (36.5 mg, 0.178 mmol) in DCM (0.22 mL) and stirred for 2 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The organic phase was separated and the aqueous phase was extracted three times with EtOAc. The combined organic phases were dried over Na2SO4, filtered, and concentrated under vacuum. The residue was dissolved in DCM and subjected to an ISCO silica column and eluted with a gradient of 0% to 100% acetone / hexane to obtain compound 31 as a white solid at 65%. Calculated mass 975.58, Found 974.69 (MH). - .

[0601] Compound 32:

[0602]

[0603] Under general conditions A1, the reaction was stirred for 2 hours to complete. Compound 32 was separated as a white solid at 21%. Calculated mass: 974.55, Measured mass: 973.25 (MH). - .

[0604] Compound 33:

[0605]

[0606] Under general conditions B1, the reaction was stirred for 0.5 h to complete. Compound 33 was separated as a white solid at 53%. Calculated mass: 988.57 g / mL; Measured mass: 987.19 g / mL (MH). - .

[0607] Compound 34:

[0608]

[0609] Tf₂O (6.0 μL, 0.0333 mmol) was added to a solution of 32-deoxyrapamycin (20.0 mg, 0.0222 mmol) and 2,6-dimethylpyridine (6.4 μL, 0.0556 mmol) in DCM (0.28 mL) at -30 °C, and the mixture was stirred for 30 min. The reaction was then heated to 0 °C and stirred for another 30 min. Then, 1-H-tetrazole (5.4 mg, 0.0778 mmol) and DIPEA (0.019 mL, 0.111 mmol) were added to the solution, and stirring was allowed at rt for 16 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO₃. The organic phase was separated, and the aqueous phase was extracted three times with EtOAc. The combined organic phases were dried over Na₂SO₄, filtered, and concentrated under vacuum. The residue was dissolved in DFM and subjected to an ISCO C18 RP column, eluted with a gradient of 50% to 100% ACN / water (+0.5% AcOH as a modifier) ​​to obtain compound 34 as a yellow solid, 69% purity. Calculated mass 951.59, Found mass 951.26 (MH). - .

[0610] Compound 35

[0611]

[0612] Under general conditions B1, the reaction was stirred for 2 days to complete. Compound 35 was separated as a white solid at 39%. Calculated mass: 967.61, Found mass: 966.88 (MH). - .

[0613] Compound 36:

[0614]

[0615] ZnCl2 (0.0584 mL, 0.0584 mmol, 1.0 M Et2O solution) was added to a solution of 40-phosphonate-32-deoxyrapamycin (19.0 mg, 0.0195 mmol) and ethanesulfonamide (10.4 mg, 0.0974 mmol) in DCM (0.39 mL). The reaction was stirred at rt for 30 min. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The organic phase was separated and the aqueous phase was extracted three times with EtOAc. The combined organic phases were dried over Na2SO4, filtered, and concentrated under vacuum. The residue was dissolved in DMF and subjected to an ISCO C18 RP column and eluted with a gradient of 50% to 90% ACN / water (+0.5% AcOH as a modifier) ​​to give compound 36 as a white solid at 29%. Calculated mass: 1050.56; Measured mass: 1049.67 (MH) - .

[0616] Compound 37:

[0617]

[0618] pTSA·H₂O (0.4 mg, 0.0021 mmol) was added to a solution of 40-dimethylphosphonate-32-deoxyrapamycin (compound 31, 20.0 mg, 0.0205 mmol) and propylsulfonamide (24.8 mg, 0.205 mmol) in ACN (0.21 mL). The reaction was stirred at rt for 2 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO₃. The organic phase was separated and the aqueous phase was extracted three times with EtOAc. The combined organic phases were dried over Na₂SO₄, filtered, and concentrated under vacuum. The residue was dissolved in DMF and subjected to an ISCO C18 RP column and eluted with a gradient of 50% to 90% ACN / water (+0.5% AcOH as a modifier) ​​to give compound 37 as a white solid at 49%. Calculated mass: 1064.58; Measured mass: 1063.76 (MH) - .

[0619] Compound 38:

[0620]

[0621] Under general conditions A1, the reaction was stirred for 2 hours to complete. Compound 38 was separated as a white solid at 20%. Calculated mass: 1026.57 g / mL; Found mass: 1025.51 g / mL (MH).

[0622] Compound 39:

[0623]

[0624] General conditions B1 were applied. ACN was used instead of DCM as the solvent. The reaction was stirred for 1 hour to complete. Compound 39 was separated as a white solid at 32%. Calculated mass 1040.59 g, Found mass 1063.85 g (M+Na) + .

[0625] Compound 40:

[0626]

[0627] Under general conditions A1, the reaction was stirred at rt for 1 h to complete. Compound 40 was separated as a white solid at 19%. Calculated mass: 976.57 g / mL; Found mass: 975.70 g / mL (MH). - .

[0628] Compound 41:

[0629]

[0630] Under general conditions A1, the reaction was modified using 10 equivalents of sulfonamide and 8 equivalents of ZnCl2. An additional 2.0 mL of ACN was added as a co-solvent. The reaction was stirred at 39°C for 2 days to complete. Compound 41 was isolated as a white solid at 4.9%. Calculated mass: 962.55; Found mass: 962.26 (MH). - .

[0631] Compound 42:

[0632]

[0633] Under general conditions A1, the procedure was modified using 20 equivalents of sulfonamide and 15 equivalents of ZnCl2. THF was used instead of DCM as the solvent. The reaction was stirred at 39°C for 5 days to complete. Compound 42 was isolated as a white solid at 5.5%. Calculated mass: 988.57; Found mass: 987.94 (MH). - .

[0634] Compound 43(3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-21-((3-chlorophenyl)amino)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methoxy- 6,8,12,14,20,26-Hexamethyl-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-Octadecyl-3H-23,27-Epoxypyrido[2,1-c][1]oxa[4]azacyclohexadecene-1,11,28,29(4H,31H)-tetraone

[0635]

[0636] 3-Chloroaniline (0.049 mL, 0.555 mmol) and TFA (0.085 mL, 1.112 mmol) were added to a stirred solution of 32-deoxyrapamycin (50 mg, 0.055 mmol) in DCM (2 mL) at -20 °C and stirred for 2 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCO C18 column, eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 8.1 mg (15%) of a white solid compound 43. 56 H 83 ClN2O 11 The calculated MS(ESI-) value for -H is 994.57, and the measured value is 994.05.

[0637] Compound 44(3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methoxy-21-((3-methoxyphenyl)amino) -6,8,12,14,20,26-hexamethyl-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-octadecylhydro-3H-23,27-epoxypyridino[2,1-c][1]oxa[4]azacyclotriacon-1,11,28,29(4H,31H)-tetraone

[0638]

[0639] At -20°C, m-aminoanisole (0.062 mL, 0.555 mmol) and TFA (0.085 mL, 1.112 mmol) were added to a stirred solution of 32-deoxyrapamycin (50 mg, 0.055 mmol) in DCM (2 mL) and stirring was continued for 2 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCOC18 column, eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 3.0 mg (5%) of the compound as a white solid. 57 H 86 N2O 12 The calculated MS(ESI-) value for -H is 990.62, and the measured value is 990.14.

[0640] Compound 45(3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-21-((1,2,5-oxadiazol-3-yl)amino)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methyl oxy-6,8,12,14,20,26-hexamethyl-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-octadecylhydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclotrimonene-1,11,28,29(4H,31H)-tetraone.

[0641]

[0642] 1,2,5-oxadiazol-3-amine (30 mg, 0.333 mmol) and TFA (0.051 mL, 3.32 mmol) were added to a stirred solution of 32-deoxyrapamycin (30 mg, 0.033 mmol) in DCM (2 mL) at -20 °C and stirred for 2 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCOC18 column, eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 9.5 mg (30%) of a white solid compound at 45 °C. 57 H 86 N2O 12 The calculated MS(ESI-) value for -H is 952.58, and the measured value is 952.03.

[0643] Compound 46. N-((3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methoxy-6,8,12,14,20,26-hexamethyl-1,11,2 8,29-Tetraoxo-1,4,5,6,9,10,11,12,13,14,21,22,23,24,25,26,27,28,29,31,32,33,34,34a-tetraco-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclotrimonen-21-yl)-3,5-dimethylisoxazole-4-sulfonamide.

[0644]

[0645] Dimethyl-1,2-oxazol-4-sulfonamide (157 mg, 0.899 mmol) and ZnCl2 (0.889 mL, 0.889 mmol) were added to a stirred solution of 32-deoxyrapamycin (80 mg, 0.089 mmol) in DCM (2 mL) at -20 °C and stirred for 2 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCO C18 column and eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 8.3 mg (9%) of a white solid compound 46.55 H 85 N3O 14 The calculated MS(ESI-) value of SH is 1043.58, and the measured value is 1042.87.

[0646] Compound 47(3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methoxy-21-((4-methoxyphenyl)amino)- 6,8,12,14,20,26-Hexamethyl-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-Octadecyl-3H-23,27-Epoxypyrido[2,1-c][1]oxa[4]azacyclotrimonene-1,11,28,29(4H,31H)-tetraone.

[0647]

[0648] p-Aminoanisole (0.064 mL, 0.556 mmol) and TFA (0.085 mL, 1.112 mmol) were added to a stirred solution of 32-deoxyrapamycin (50 mg, 0.055 mmol) in DCM (2 mL) at -20 °C, and stirring was continued for 4 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCOC18 column, eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 2.0 mg (4%) of white solid. 57 H 86 N2O 12 The calculated MS(ESI-) value for -H is 991.32, and the measured value is 990.67.

[0649] Compound 48. (3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-21-((4-chlorophenyl)amino)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methoxy-6,8 ,12,14,20,26-hexamethyl-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-octadecylhydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclotrimonene-1,11,28,29(4H,31H)-tetraone.

[0650]

[0651] 4-Chloroaniline (0.050 mL, 0.556 mmol) and TFA (0.085 mL, 1.112 mmol) were added to a stirred solution of 32-deoxyrapamycin (50 mg, 0.055 mmol) in DCM (2 mL) at -20 °C and stirred for 2 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCO C18 column, eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 5.2 mg (9%) of white solid. 56 H 83 ClN2O 11 The calculated MS(ESI-) value for -H is 994.57, and the measured value is 993.86.

[0652] Compound 49(3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methoxy-6,8,12,14,20,2 6-Hexamethyl-21-(p-Tolylamino)-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-octadecylhydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohexadecene-1,11,28,29(4H,31H)-tetraone.

[0653]

[0654] p-Toluidine (0.060 mL, 0.556 mmol) and TFA (0.085 mL, 1.112 mmol) were added to a stirred solution of 32-deoxyrapamycin (50 mg, 0.055 mmol) in DCM (2 mL) at -20 °C and stirred for 2 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCO C18 column, eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 1.6 mg (3%) of white solid. 57 H 86 N2O 11 The calculated MS(ESI-) value for -H is 974.62, and the measured value is 973.76.

[0655] Compound 50(3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methoxy-6,8,12,14,20,2 6-Hexamethyl-21-(m-Tolylamino)-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-octadecylhydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohexadecene-1,11,28,29(4H,31H)-tetraone.

[0656]

[0657] To a stirred solution of 32-deoxyrapamycin (50 mg, 0.055 mmol) in DCM (2 mL), m-toluidine (0.060 mL, 0.556 mmol) and TFA (0.085 mL, 1.112 mmol) were added at -20 °C and stirred for 2 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCO C18 column, eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 2.5 mg (5%) of white solid. 57 H 86 N2O11 The calculated MS(ESI-) value for -H is 974.62, and the measured value is 973.95.

[0658] Compound 51

[0659] N-((3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methoxy-6,8,12,14,20,26-hexamethyl-1,11 ,28,29-Tetraoxo-1,4,5,6,9,10,11,12,13,14,21,22,23,24,25,26,27,28,29,31,32,33,34,34a-tetrahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclotrimonen-21-yl)-5-methylisoxazole-4-sulfonamide.

[0660]

[0661] 5-Methyl-1,2-oxazol-4-sulfonamide (144 mg, 0.889 mmol) and ZnCl2 (0.889 mL, 0.889 mmol) were added to a stirred solution of 32-deoxyrapamycin (80 mg, 0.089 mmol) in DCM (2 mL) and stirred for 3 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCO C18 column and eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 4.1 mg (9%) of white solid. 54 H 83 N3O 14 The calculated MS(ESI-) value of SH is 1030.33, and the measured value is 1029.16.

[0662] Compound 52(3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methoxy-6,8,12,14,20,2 6-Hexamethyl-21-(p-Tolylamino)-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-octadecylhydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohexadecene-1,11,28,29(4H,31H)-tetraone.

[0663]

[0664] Aminopyrazine (53 mg, 0.556 mmol) and TFA (0.085 mL, 1.112 mmol) were added to a stirred solution of 32-deoxyrapamycin (50 mg, 0.055 mmol) in DCM (2 mL) at -20 °C and stirred for 2 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCO C18 column, eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 4.2 mg (8%) of white solid. 54 H 82 N4O 11 The calculated MS(ESI-) value for -H is 962.60, and the measured value is 961.91.

[0665] Compound 53

[0666] N-((3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methoxy-6,8,12,14,20,26-hexamethyl-1,11 ,28,29-Tetraoxo-1,4,5,6,9,10,11,12,13,14,21,22,23,24,25,26,27,28,29,31,32,33,34,34a-tetrahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclotrimonen-21-yl)-3-methylisoxazole-4-sulfonamide.

[0667]

[0668] 3-Methyl-1,2-oxazol-4-sulfonamide (144 mg, 0.889 mmol) and ZnCl2 (0.889 mL, 0.889 mmol) were added to a stirred solution of 3,2-deoxyrapamycin (80 mg, 0.089 mmol) in DCM (2 mL) and stirred for 3 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCO C18 column and eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 1.7 mg (2%) of white solid. 54 H 83 N3O 14 The calculated MS(ESI-) value of SH is 1030.33, and the measured value is 1029.27.

[0669] Compound 54(3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methoxy-6,8,12,14,20,26-hexamethylene 21-((3-(trifluoromethyl)phenyl)amino)-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-octadecylhydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohexadecene-1,11,28,29(4H,31H)-tetraone.

[0670]

[0671] 3-Trifluoromethylaniline (0.065 mL, 0.556 mmol) and TFA (0.085 mL, 1.112 mmol) were added to a stirred solution of 32-deoxyrapamycin (50 mg, 0.055 mmol) in DCM (2 mL) at -20 °C and stirred for 2 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCO C18 column, eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 6.5 mg (11%) of white solid. 57 H 83 F3N2O 11 The calculated MS(ESI-) value for -H is 1028.57, and the measured value is 1027.90.

[0672] Compound 55(3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methoxy-6,8,12,14,20,26-hexamethylene 21-((4-(trifluoromethyl)phenyl)amino)-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-octadecylhydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohexadecene-1,11,28,29(4H,31H)-tetraone.

[0673]

[0674] 4-Trifluoromethylaniline (0.065 mL, 0.556 mmol) and TFA (0.085 mL, 1.112 mmol) were added to a stirred solution of 32-deoxyrapamycin (50 mg, 0.055 mmol) in DCM (2 mL) at -20 °C and stirred for 2 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCO C18 column, eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 2.3 mg (4%) of white solid. 57 H 83 F3N2O11 The calculated MS(ESI-) value for -H is 1028.57, and the measured value is 1028.05.

[0675] Compound 56(4-(((3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methoxy-6,8,12,14,20,26-hexadecyl Methyl-1,11,28,29-tetraoxo-1,4,5,6,9,10,11,12,13,14,21,22,23,24,25,26,27,28,29,31,32,33,34,34a-tetrahydro-3H-23,27-epoxypyridino[2,1-c][1]oxa[4]azacyclohexaeno-21-yl)amino)benzonitrile)

[0676]

[0677] 4-Aminobenzonitrile (0.065 mL, 0.556 mmol) and TFA (0.085 mL, 1.112 mmol) were added to a stirred solution of 32-deoxyrapamycin (50 mg, 0.055 mmol) in DCM (8 mL) at -20 °C, and the reaction was stirred for 3 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCO C18 column, eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 2.4 mg (4%) of white solid. 57 H 83 N3O 11 The calculated MS(ESI-) value for -H is 985.60, and the measured value is 985.08.

[0678] Compound 57(3-(((3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methoxy-6,8,12,14,20,26- Hexamethyl-1,11,28,29-tetraoxo-1,4,5,6,9,10,11,12,13,14,21,22,23,24,25,26,27,28,29,31,32,33,34,34a-tetrahydro-3H-23,27-epoxypyridino[2,1-c][1]oxa[4]azacyclohexane-21-yl)amino)benzonitrile

[0679]

[0680] 3-Aminobenzonitrile (0.065 mL, 0.556 mmol) and TFA (0.085 mL, 1.112 mmol) were added to a stirred solution of 32-deoxyrapamycin (75 mg, 0.083 mmol) in DCM (10 mL) at -20 °C and stirred for 3 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCOC18 column, eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 3.1 mg (6%) of white solid. 57 H 83 N3O 11 The calculated MS(ESI-) value for -H is 984.60, and the measured value is 985.01.

[0681] Compound 58((3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methoxy-6,8,12,14,20,26-hexamethylene 21-((3-(trifluoromethoxy)phenyl)amino)-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-octadecylhydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohexadecene-1,11,28,29(4H,31H)-tetraone)

[0682]

[0683] 3-Trifluoromethoxyaniline (0.076 mL, 0.834 mmol) and TFA (0.128 mL, 1.667 mmol) were added to a stirred solution of 32-deoxyrapamycin (50 mg, 0.055 mmol) in DCM (10 mL) at -20 °C and stirred for 3 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCO C18 column, eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 10.9 mg (13%) of white solid. 57 H 83 F3N2O 12 The calculated MS(ESI-) value for -H is 1043.59, and the measured value is 1044.09.

[0684] Compound 59((3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-21-((3-ethoxyphenyl)amino)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methoxy -6,8,12,14,20,26-hexamethyl-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-octadecylhydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclotriacontria-1,11,28,29(4H,31H)-tetraone)

[0685]

[0686] 3-ethoxyaniline (0.095 mL, 0.834 mmol) and TFA (0.128 mL, 1.667 mmol) were added to a stirred solution of 32-deoxyrapamycin (75 mg, 0.083 mmol) in DCM (10 mL) at -20 °C and stirred for 3 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCOC18 column, eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 6.5 mg (8%) of white solid. 58 H 88 N2O 12 The calculated MS(ESI-) value for -H is 1003.63, and the measured value is 1003.97.

[0687] Compound 60((3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methoxy-6,8,12,14,20,26 -hexamethyl-21-(thiophen-3-ylamino)-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-octadecylhydro-3H-23,27-epoxypyridino[2,1-c][1]oxa[4]azacyclohexadecene-1,11,28,29(4H,31H)-tetraone)

[0688]

[0689] Thiophene-3-amine hydrochloride (113 mg, 0.834 mmol) and TFA (0.128 mL, 1.667 mmol) were added to a stirred solution of 32-deoxyrapamycin (75 mg, 0.083 mmol) in DCM (10 mL) at -20 °C and stirred for 3 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCOC18 column, eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 2.0 mg (2%) of white solid. 54 H 82 N2O 11The calculated MS(ESI-) value of SH is 965.56, and the measured value is 965.86.

[0690] Compound 61(N-((3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methoxy-6,8,12,14,20,26-hexamethyl -1,11,28,29-tetraoxo-1,4,5,6,9,10,11,12,13,14,21,22,23,24,25,26,27,28,29,31,32,33,34,34a-tetraco-3H-23,27-epoxypyridino[2,1-c][1]oxa[4]azacyclohexane-21-yl)isoxazole-4-sulfonamide)

[0691]

[0692] 1,2-oxazol-4-sulfonamide (124 mg, 0.834 mmol) and ZnCl2 (0.834 mL, 0.834 mmol) were added to a stirred solution of 32-deoxyrapamycin (75 mg, 0.083 mmol) in DCM (10 mL) and stirred for 16 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCO C18 column, eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 1.0 mg (1%) of white solid. 53 H 81 N3O 14 The calculated MS(ESI-) value of SH is 1015.54, and the measured value is 1015.14.

[0693] Compound 62((3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methoxy-21-((3-methoxyphenyl)(methyl)amino) (6,8,12,14,20,26-hexamethyl-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-octadecylhydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohexadecene-1,11,28,29(4H,31H)-tetraone)

[0694]

[0695] 3-Methoxy-N-methylaniline (0.094 mL, 0.834 mmol) and TFA (0.128 mL, 1.667 mmol) were added to a stirred solution of 32-deoxyrapamycin (75 mg, 0.083 mmol) in DCM (10 mL) at -20 °C and stirred for 6 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCO C18 column, eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 5.8 mg (7%) of white solid. 58 H 88 N2O 12 The calculated MS(ESI-) value for -H is 1004.63, and the measured value is 1004.39.

[0696] Compound 63((3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methoxy-6,8,12,14,20,26 -Hexamethyl-21-(methyl(phenyl)amino)-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-octadecylhydro-3H-23,27-epoxypyridino[2,1-c][1]oxa[4]azacyclohexadecene-1,11,28,29(4H,31H)-tetraone)

[0697]

[0698] N-methylaniline (0.120 mL, 1.112 mmol) and TFA (0.170 mL, 2.223 mmol) were added to a stirred solution of 32-deoxyrapamycin (100 mg, 0.111 mmol) in DCM (10 mL) at -20 °C and stirred for 6 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCOC18 column, eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 2.6 mg (3%) of white solid. 57 H 86 N2O 11 The calculated MS(ESI-) value for -H is 974.62, and the measured value is 974.07.

[0699] Compound 64(3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methoxy-6,8,12,14,20, 26-Hexamethyl-21-(methyl(phenyl)amino)-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-octadecylhydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohexadecene-1,11,28,29(4H,31H)-tetraone:

[0700]

[0701] N-methylaniline (0.240 mL, 2.223 mmol) and TFA (0.340 mL, 4.445 mmol) were added to a stirred solution of 32-deoxyrapamycin (200 mg, 0.222 mmol) in DCM (10 mL) at -20 °C and stirred for 16 h to rt. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCO C18 column and eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 4.1 mg (2%) of white solid.57 H 86 N2O 11 The calculated MS(ESI-) value for -H is 974.62, and the measured value is 974.33.

[0702] Compound 65(3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-21-(3,4-dihydroquinoline-1(2H)-yl)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methyl oxy-6,8,12,14,20,26-hexamethyl-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-octadecylhydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohexadecene-1,11,28,29(4H,31H)-tetraone:

[0703]

[0704] 1,2,3,4-Tetrahydroquinoline (0.278 mL, 2.223 mmol) and TFA (0.340 mL, 4.445 mmol) were added to a stirred solution of 32-deoxyrapamycin (200 mg, 0.222 mmol) in DCM (10 mL) at -20 °C and stirred for 16 h to rt. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCO C18 column and eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 4.8 mg (2%) of white solid. 57 H 86 N2O 11 The calculated MS(ESI-) value for -H is 1000.64, and the measured value is 1000.17.

[0705] Compound 66(3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,21R,23S,26R,27R,34S)-21-(1,1-dioxaisothiazolin-2-yl)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10- Methoxy-6,8,12,14,20,26-hexamethyl-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-octadecylhydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohexadecene-1,11,28,29(4H,31H)-tetraone:

[0706]

[0707] 1,3-propanesulfonamide (0.136 mL, 1.112 mmol) and ZnCl2 (1.112 mL, 1.112 mmol) were added to a stirred solution of 3,2-deoxyrapamycin (100 mg, 0.111 mmol) in DCM (10 mL) and stirred for 16 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCO C18 column, eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 14.4 mg (13%) of white solid. 53 H 84 N2O 13 The calculated MS(ESI-) value of SH is 988.57, and the measured value is 988.16.

[0708] Compound 67: (3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,21R,23S,26R,27R,34S)-21-(1,1-isothiazolidin-2-yl)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10 -Methoxy-6,8,12,14,20,26-hexamethyl-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-octadecylhydro-3H-23,27-epoxypyridino[2,1-c][1]oxa[4]azacyclotrimonene-1,11,28,29(4H,31H)-tetraone

[0709]

[0710] 1,3-propanesulfonamide (0.136 mL, 1.112 mmol) and ZnCl2 (1.112 mL, 1.112 mmol) were added to a stirred solution of 3,2-deoxyrapamycin (100 mg, 0.111 mmol) in DCM (10 mL) and stirred for 16 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCO C18 column and eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 8.5 mg (8%) of white solid. 53 H 84 N2O 13 The calculated MS(ESI-) value of SH is 988.57, and the measured value is 988.01.

[0711] Compound 68: (3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-21-((2-hydroxyethyl)(phenyl)amino)-10-methoxy 6,8,12,14,20,26-hexamethyl-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-octadecylhydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohexadecene-1,11,28,29(4H,31H)-tetraone:

[0712]

[0713] 2-(phenylamino)ethanol (0.122 mL, 0.834 mmol) and TFA (0.128 mL, 1.667 mmol) were added to a stirred solution of 32-deoxyrapamycin (75 mg, 0.083 mmol) in DCM (10 mL) at -20 °C and stirred for 16 h to rt. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCO C18 column and eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 15.5 mg (19%) of white solid. MS (ESI-) calculated value of C58H88N2O12-H = 1004.63, found value 1004.08.

[0714] Compound 69: (3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methoxy-21-((2-methoxyethyl)(phenyl)amino) (6,8,12,14,20,26-hexamethyl-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-octadecylhydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohexadecene-1,11,28,29(4H,31H)-tetraone:

[0715]

[0716] N-(2-methoxyethyl)aniline (0.125 mL, 0.834 mmol) and TFA (0.128 mL, 1.667 mmol) were added to a stirred solution of 32-deoxyrapamycin (75 mg, 0.083 mmol) in DCM (10 mL) at -20 °C and stirred for 16 h to rt. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCO C18 column and eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 2.7 mg (3%) of white solid. MS (ESI-) calculated value of C59H90N2O12-H = 1018.65, found value 1018.21.

[0717] Compound 70: (3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-21-((3-(difluoromethoxy)phenyl)amino)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methyl oxy-6,8,12,14,20,26-hexamethyl-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-octadecylhydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohexadecene-1,11,28,29(4H,31H)-tetraone:

[0718]

[0719] 3-Difluoromethoxyaniline (0.139 mL, 1.112 mmol) and TFA (0.170 mL, 2.223 mmol) were added to a stirred solution of 32-deoxyrapamycin (100 mg, 0.111 mmol) in DCM (10 mL) at -20 °C and stirred at room temperature for 16 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCO C18 column and eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 7.7 mg (7%) of white solid. MS (ESI-) calculated value of C57H84F2N2O12-H = 1026.60, found value 1026.16.

[0720] Compound 71

[0721]

[0722] (1R,2R,4S)-4-((2R)-2-((3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-9,27-dihydroxy-10-methoxy-21-((2-methoxyethyl)(phenyl)amino)-6,8,12,14,20,26-hexamethyl-1,11,28 ,29-tetraoxo-1,4,5,6,9,10,11,12,13,14,21,22,23,24,25,26,27,28,29,31,32,33,34,34a-tetrahydro-3H-23,27-epoxypyridino[2,1-c][1]oxa[4]azacyclohexane-3-yl)propyl)-2-methoxycyclohexyl dimethylphosphines.

[0723] Compound 71 was prepared using compound 31 as a starting material in the same manner as compound 69.

[0724] Compound 72

[0725] N-((3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methoxy-6,8,12,14,20,26-hexamethyl-1, 11,28,29-Tetraoxo-1,4,5,6,9,10,11,12,13,14,21,22,23,24,25,26,27,28,29,31,32,33,34,34a-tetrahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohexane-21-yl)-3-methylbenzenesulfonamide.

[0726]

[0727] 3-Methylbenzene-1-sulfonamide (95 mg, 0.555 mmol) and ZnCl2 (0.56 mL, 0.555 mmol) were added to a stirred solution of 32-deoxyrapamycin (50 mg, 0.055 mmol) in DCM (6 mL) and stirred for 16 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCOC18 column and eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 4.2 mg (7%) of a yellow solid, RGN-3768. MS (ESI-) calculated value of C57H86N2O13S-H = 1038.59, found value 1038.62.

[0728] Compound 73

[0729] N-((3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methoxy-6,8,12,14,20,26-hexamethyl-1, 11,28,29-Tetraoxo-1,4,5,6,9,10,11,12,13,14,21,22,23,24,25,26,27,28,29,31,32,33,34,34a-tetrahydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclohexane-21-yl)-N-methylbenzenesulfonamide.

[0730] N-methylbenzenesulfonamide (0.080 mL, 0.555 mmol) and ZnCl2 (0.56 mL, 0.555 mmol) were added to a stirred solution of 32-deoxyrapamycin (50 mg, 0.055 mmol) in DCM (6 mL) and stirring was continued for 16 h. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCOC18 column and eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 26.9 mg (52%) of a yellow solid, RGN-3769. MS (ESI-) calculated value of C57H86N2O13S-H = 1038.59, found value 1038.24.

[0731]

[0732] Compound 74. (3S,6S,7E,9R,10R,12R,14S,15E,17E,19E,23S,26R,27R,34aS)-21-((3-fluorophenyl)amino)-9,27-dihydroxy-3-((R)-1-((1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl)propyl-2-yl)-10-methoxy-6,8 ,12,14,20,26-hexamethyl-5,6,9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-octadecylhydro-3H-23,27-epoxypyrido[2,1-c][1]oxa[4]azacyclotrimonene-1,11,28,29(4H,31H)-tetraone.

[0733]

[0734] 3-Fluoroaniline (0.322 mL, 3.335 mmol) and TFA (0.510 mL, 6.670 mmol) were added to a stirred solution of 32-deoxyrapamycin (300 mg, 0.333 mmol) in DCM (10 mL) at -20 °C and stirred for 16 h to rt. The resulting mixture was diluted with EtOAc and quenched with a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was dissolved in DMF and subjected to an ISCO C18 column and eluted with 60%–80% ACN / H2O. The purified product fraction was lyophilized to give 2.9 mg (1%) of white solid 74. MS (ESI-) calculated value of C56H83FN2O11-H = 978.60, found value 978.85.

[0735] Example 3. Inhibition of wild-type (WT) 293T cells and FKBP12 knockout 293T cells by representative compounds

[0736] Example: Generation of FKBP 12 knockout cells. A CRISPR / Cas9 system was used to deliver ribonucleoprotein complexes containing guide RNA sequences targeting FKBP12 (acCGGTGTAGTGCACCACGC, position 1369062; ccactactcacCGTCTCCTG, position 1369181) into HEK293T cells using Lipofectamine CRISPRMAX transfection reagent (Invitrogen CMAX00008). Cell clones were selected by the following: Western blotting with an anti-FKBP specific antibody (Novus, NB300-508). Single cells deficient in the FKBP12 protein were selected and cloned.

[0737] Wild-type (WT) 293T cells and FKBP12 knockout 293T cells were treated with RAD001 (everolimus) and the test compound. WT 293T cells and FKBP12 knockout 293T cells were cultured in Dulbecco modified Eagle medium (Gibco #11971-025) supplemented with 10% fetal bovine serum (Gibco #26140-079). Cells were seeded at a density of 15,000 cells / well in poly-D-lysine-coated 96-well plates (Corning, #354461) and incubated at 37°C, 5% CO2 for 24 hours until they reached ~80% confluence. Cells were treated with RAD001 or the test compound reconstituted in dimethyl sulfoxide (DMSO) at doses ranging from 8 to 11 points (from 0.001 pM to 10 μM). All treatments were performed in duplicate at 37°C for 2 hours. Cell culture medium supplemented with blank dimethyl sulfoxide (DMSO) was used as a negative control for all compounds. S6K1 (Thr389) phosphorylation was measured using an ELISA kit (Invitrogen 85-86052-11) according to the manufacturer's protocol.

[0738] S6K1 is a close downstream target of mTORC1, and therefore phosphorylation of S6K1 at Thr389 residues was used to measure mTORC1 activity (PMID: 16968213). The amount of phosphorylated S6K1 (Thr389) was measured using RAD001 or compound 44 ( Figure 1A and Figure 1B ) and compound 45 ( Figure 1C and Figure 1D Measurements were taken in WT 293T cells treated with 10000 and FKBP12 knockout 293T cells.

[0739] In WT cells, both RAD001 and compound 44 completely inhibited S6K1 phosphorylation in a dose-dependent manner. Figure 1AIn WT cells, RAD001 inhibited 50% (IC50) of S6K1 phosphorylation at 0.66 nM, while compound 44 inhibited 50% (IC50) of S6K1 phosphorylation at 11 nM. Therefore, in WT cells, compound 44 was ~17 times less potent than RAD001. Figure 1A ).

[0740] In FKBP12 knockout cells (i.e., cells lacking FKBP12), RAD001 achieved ~30% inhibition of S6K1 phosphorylation at 100 nM, while compound 44 achieved ~30% inhibition of S6K1 phosphorylation at 10 μM. Therefore, in FKBP12KO cells, at the highest tested concentration of 10 μM, compound 44 was ~100-fold less potent than RAD001. Figure 1B In FKBP12 knockout cells, compound 44 did not inhibit S6K1 phosphorylation at any tested concentration below 10 μM. Figure 1B These data demonstrate that compound 44 is relatively more selective for FKBP12 compared to RAD001, and that FKBP12 is required to inhibit mTORC1 signaling when used in this cellular system at concentrations of 1 μM or lower.

[0741] In WT cells, both RAD001 and compound 45 completely inhibited S6K1 phosphorylation in a dose-dependent manner. Figure 1C In WT cells, RAD001 inhibited 50% (IC50) of S6K1 phosphorylation at 0.47 nM, while compound 45 inhibited 50% (IC50) of S6K1 phosphorylation at 8.4 nM. Therefore, in WT cells, compound 45 was ~18 times less potent than RAD001. Figure 1C ).

[0742] In FKBP12 knockout cells (i.e., cells lacking FKBP12), RAD001 at a concentration of 100 nM achieved ~35% inhibition of S6K1 phosphorylation, and at the highest tested concentration of 10 μM, RAD001 inhibited S6K1 phosphorylation by ~80%. In FKBP12 knockout cells, compound 45 at a concentration of 10 μM inhibited S6K1 phosphorylation by ~25%. Figure 1D RAD001 achieved a similar level of S6K1 inhibition at ~100 nM. Therefore, in FKBP12 knockout cells, compound 45 was ~100 times less potent than RAD001 when tested at a concentration of 10 μM. Figure 1D ).

[0743] These data demonstrate that compound 45 is relatively more selective for FKBP12 compared to RAD001.

[0744] In WT HEK293T cells expressing FKBP12, both RAD001 and compound 18 achieved complete inhibition of S6K1 (Thr389) phosphorylation. Figure 2A In WT cells, compound 18 was ~9-fold less potent than RAD001. However, in the absence of FKBP12 (in FKBP12 knockout cells), RAD001 achieved ~40% S6K1 (Thr389) inhibition at 447 nM, while compound 18 achieved a similar level of S6K1 (Thr389) inhibition at 9.5 μM. Figure 2B This indicates that in FKBP12 knockout cells, compound 18 is approximately 20 times less potent than RAD001. These data demonstrate that compound 18 is relatively more selective for FKBP12 compared to RAD001; however, at the highest tested concentration, in the absence of FKBP12, compound 18 still achieves approximately 40% inhibition of S6K1.

[0745] In WT HEK293T cells expressing FKBP12, both RAD001 and compound 16 achieved complete inhibition of S6K1 (Thr389) phosphorylation. Figure 3A In WT cells, compound 16 was ~28 times less potent than RAD001. However, in the absence of FKBP12 (in FKBP12 knockout cells), compound 16 was as potent as RAD001. Figure 3B At the highest concentration tested (10 μM), both rapamycin analogs achieved ~70% inhibition of S6K1 (Thr389) phosphorylation. These data suggest that compound 16 may have a lower affinity for FKBP12 compared to RAD001, however, it maintains affinity for other FKBPs expressed in HEK293T cells.

[0746] In FKBP12-expressing WT HEK293T cells, RAD001, compound 26, and compound 28 achieved complete inhibition of S6K1 (Thr389) phosphorylation. Figure 4ACells were treated with the following compounds for 2 hours: RAD001 (everolimus; dashed circle, IC50 0.017 nM), compound 26 (solid square, IC50 0.55 nM), and compound 28 (solid cross, IC50 3.0 nM). In FKBP12 knockout 293T cells, while RAD001 (everolimus; dashed circle, IC50 31.3 nM) achieved approximately 50% S6K1 inhibition, compound 26 (solid square, IC50 unmeasurable / undetectable) did not inhibit S6K1 at any tested concentration, indicating that compound 26 is more selective for FKBP12 compared to RAD001. In FKBP12 KO cells, compound 28 (solid line with a cross, IC50 unmeasurable / undetectable) behaved in a similar manner to RAD001, indicating that compound 28 did not have higher selectivity for FKBP12 compared to RAD001.

[0747] In wild-type HEK293T cells expressing FKBP12, RAD001 (IC50 = 0.319 nM), along with compounds 47 (IC50 = 69 nM), 48 (IC50 = 9.8 nM), and 49 (IC50 = 9.5 nM), inhibited mTORC1 signaling in a dose-dependent manner, achieving near-complete inhibition of S6K1 (Thr389) phosphorylation with increasing concentration. Figure 5A , Figure 5C and Figure 5E The new compounds are 219 times (47), 30 times (48) and 29 times (49) less potent than RAD001, respectively, as indicated by their corresponding IC50 values.

[0748] In the absence of FKBP12, RAD001 inhibited S6K1 (Thr389) phosphorylation in FKBP12 knockout cells, achieving 42% inhibition at a concentration of 10 μM. New compounds 47, 48, and 49, tested in parallel with RAD001, did not inhibit S6K1 (Thr389) phosphorylation at any of the tested concentrations. Figure 5B , Figure 5D and Figure 5F These results indicate that, unlike RAD001, the activities of compounds 47, 48, and 49 are dependent on FKBP12, and that these compounds do not inhibit mTORC1 signaling in the absence of FKBP12, meaning that compounds 47, 48, and 49 are relatively more selective for FKBP12 compared to RAD001.

[0749] In wild-type HEK293T cells expressing FKBP12, RAD001 (IC50 = 0.365 nM), along with compounds 50 (IC50 = 5.72 nM), 51 (IC50 = 4.53 nM), and 52 (IC50 = 9.15 nM), inhibited mTORC1 signaling in a dose-dependent manner, achieving near-complete inhibition of S6K1 (Thr389) phosphorylation with increasing concentration. Figure 6A , Figure 6C and Figure 6E These new compounds are 15 times (50), 12 times (51) and 25 times (52) less potent than RAD001, respectively, as indicated by their corresponding IC50 values.

[0750] In the absence of FKBP12, RAD001 inhibited S6K1 (Thr389) phosphorylation in FKBP12 knockout cells in a dose-dependent manner, achieving 45% inhibition at 10 μM. New compounds 50 and 52, tested in parallel with RAD001, showed only slight inhibition of S6K1 (Thr389) phosphorylation at the highest tested concentration (12% inhibition in the case of compound 50), or no inhibition at any tested concentration (see compound 52). Figure 6B , Figure 6F These results indicate that, unlike RAD001, the activity of compounds 50 and 52 is dependent on FKBP12, and that these compounds do not inhibit mTORC1 signaling in the absence of FKBP12, meaning that compounds 50 and 52 are selective for FKBP12.

[0751] In FKBP12 knockout cells, compound 51 retained some potency and inhibited S6K1(Thr389) phosphorylation by 18% at a concentration of 1 μM, with no further inhibition at 10 μM. Figure 6D In the same assay, RAD001 achieved 20% inhibition of S6K1(Thr389) phosphorylation at a concentration of 115 nM. These results indicate that compound 51 is not completely selective for FKBP12 and that the inhibitory effect of compound 51 on mTORC1 signaling can be mediated by FKBP other than FKBP12.

[0752] In wild-type HEK293T cells expressing FKBP12, RAD001 (IC50 = 0.084 nM), along with compounds 57 (IC50 = 3.68 nM), 58 (IC50 = 2.94 nM), and 59 (IC50 = 0.17 nM), inhibited mTORC1 signaling in a dose-dependent manner, achieving near-complete inhibition of S6K1 (Thr389) phosphorylation at the highest tested concentration. Figure 7A , Figure 7C and Figure 7E In this assay, the potency of the new compounds was 44 times (57), 35 times (58) and 2 times (59) lower than that of RAD001, respectively, as indicated by their corresponding IC50 values.

[0753] In the absence of FKBP12, RAD001 and all other tested compounds (57, 58, and 59) inhibited mTORC1 signaling in a dose-dependent manner in FKBP12 knockout cells. Figure 7B , Figure 7D and Figure 7F These results indicate that, similar to RAD001, compounds (57, 58, and 59) retain their potency in the absence of FKBP12, meaning that their inhibitory effect on mTORC1 signaling can be mediated by FKBP other than FKBP12, i.e., compounds 57, 58, and 59 are not relatively FKBP12 selective.

[0754] In wild-type HEK293T cells expressing FKBP12, both RAD001 (IC50 = 0.235 nM) and compound 63 (IC50 = 2.43 nM) inhibited mTORC1 signaling in a dose-dependent manner, and achieved almost complete inhibition of S6K1 (Thr389) phosphorylation with increasing concentration. Figure 8A In this assay, the potency of 63 was 10 times lower than that of RAD001, as indicated by the corresponding IC50 value.

[0755] In FKBP12 knockout cells, compound 63 retained some potency and inhibited S6K1(Thr389) phosphorylation by 17% at the maximum tested concentration of 10 μM. Figure 8BIn the same assay, RAD001 achieved 20% inhibition of S6K1(Thr389) phosphorylation at a concentration of 1.3 nM and a further 58% inhibition at 10 μM. These results indicate that even though compound 63 is not specifically selective for FKBP12, it is still more selective than RAD001. In wild-type cells expressing FKBP12, 63 was 10-fold less potent than RAD001; however, in FKBP12 knockout cells, 63 did not achieve more than 17% inhibition of S6K1(Thr389), while RAD001 inhibited S6K1(Thr389) by up to 58% in a dose-dependent manner.

[0756] In wild-type HEK293T cells expressing FKBP12, both RAD001 (IC50 = 0.389 nM) and compound 69 (IC50 = 1.53 nM) inhibited mTORC1 signaling, and with increasing concentration, achieved almost complete inhibition of S6K1 (Thr389) phosphorylation. Figure 9A Of these, 69 is 4 times less effective than RAD001, as indicated by the corresponding IC50 value.

[0757] In the absence of FKBP12, increasing the concentration of RAD001 in FKBP12 knockout cells achieved up to 73.5% dose-dependent inhibition of S6K1(Thr389) phosphorylation at the highest tested concentration (10 μM). In the absence of FKBP12, compound 69 retained very low potency and achieved approximately 30% inhibition of S6K1(Thr389) phosphorylation at the highest tested concentration (10 μM). In the same FKBP12 knockout cell assay, RAD001 achieved 30% inhibition of S6K1(Thr389) phosphorylation at a concentration of ~3.0 nM. These results indicate that even though compound 69 is not specifically selective for FKBP12, it is still more selective than RAD001. In wild-type cells expressing FKBP12, compound 69 was only 4-fold less potent than RAD001. However, in FKBP12 knockout cells, compound 69 inhibited S6K1 phosphorylation by 30% at a concentration approximately 3000 times higher than that of RAD001.

[0758] In wild-type HEK293T cells expressing FKBP12, both RAD001 (IC50 = 0.174 nM) and compound 72 (IC50 = 0.914 nM) inhibited mTORC1 signaling, and with increasing concentration, achieved almost complete inhibition of S6K1 (Thr389) phosphorylation. Figure 10A In this assay, the potency of 72 was 5.2 times lower than that of RAD001, as indicated by the corresponding IC50 value.

[0759] In the absence of FKBP12, increasing the concentration of RAD001 in FKBP12 knockout cells achieved dose-dependent inhibition of S6K1(Thr389) phosphorylation, with up to 62% inhibition achieved at the highest tested concentration (10 μM). Figure 10B In the absence of FKBP12, compound 72 also retained some of its potency and inhibited S6K1(Thr389) phosphorylation in a dose-dependent manner, achieving 40% inhibition at the highest tested concentration (10 μM). Figure 10B These results indicate that, although 72 is less potent than RAD001 in FKBP12 knockout cells, both rapamycin analogs behave in a similar manner. That is, when combined with FKBP other than FKBP12, they can induce mTORC1 inhibition.

[0760] In wild-type HEK293T cells expressing FKBP12, both RAD001 (IC50 = 0.174 nM) and compound 73 (IC50 = 2.22 nM) inhibited mTORC1 signaling, and with increasing concentration, achieved almost complete inhibition of S6K1 (Thr389) phosphorylation. Figure 11A In this assay, the potency of 73 was 13 times lower than that of RAD001, as indicated by the corresponding IC50 value.

[0761] In the absence of FKBP12, such as in FKBP12 knockout cells, increasing the concentrations of RAD001 and 73 achieved dose-dependent inhibition of S6K1 (Thr389) phosphorylation, with up to ~60% inhibition achieved at the highest tested concentration (10 μM). Figure 11B Furthermore, although 73 was 13 times less potent than RAD001 in wild-type cells expressing FKBP12, it achieved greater potency than RAD001 in the absence of FKBP12 (in FKBP12 knockout cells). Specifically, in FKBP12 knockout cells, 73 inhibited S6K1(Thr389) phosphorylation by 20% at a concentration of 2.7 nM, while RAD001 achieved 20% inhibition at 7.4 nM. Figure 11B However, at the highest tested concentration of 10 μM, both RAD001 and compound 73 inhibited S6K1 phosphorylation by ~60%. Figure 11B ).

[0762] These results indicate that 73 can induce mTORC1 inhibition when it is combined with FKBP other than FKBP12, meaning that 73 is not FKBP12 selective. Furthermore, the higher potency of 73 relative to RAD001 in FKBP12 knockout cells (at doses less than 1 μM) suggests that 73 may have acquired an affinity for FKBP other than FKBP12 compared to RAD001.

[0763] Example 4. Microparticle stability

[0764] Materials and methods

[0765] Buffer: PBK buffer: 50mM potassium phosphate buffer (PPB), pH 7.2, containing 3.3mM magnesium chloride.

[0766] Compound dilution:

[0767] TA intermediate solution: Dilute the compound or control (3 μL) from the stock solution (10 mM) with 297 μL of 90.0% methanol / water. (Concentration: 100 μM; 1.0% DMSO, 89.1% MeOH).

[0768] Control mixture intermediate solution: Dilute each control (3 μL) with 291 μL of 90.0% methanol / water.

[0769] Diluted TA intermediate and PC control stock solutions were prepared by transferring 80 μL of 100 μM TA stock solution to a new tube and then adding 720 μL of 50 mM potassium phosphate buffer containing 1.0% DMSO (concentration 10 μM; 1.0% DMSO, 8.91% MeOH).

[0770] Preparation of liver mixture and NADPH cofactor (described in Table 1)

[0771] Preparation of working solution for liver microsomes (1.25x) (final concentration: 0.5 mg / mL).

[0772] Table 1. Preparation of liver mixture and NADPH cofactor

[0773]

[0774] Stock solution:

[0775] Acetonitrile:methanol (9:1, v / v) (including 200 ng / mL tolbutamide and 200 ng / mL labetalol hydrochloride as internal standards).

[0776] Working solution: Dilute the intermediate solution (80 μL) with 720 μL of 50 mM potassium phosphate buffer containing 1.0% DMSO (concentration: 10 μM; 1.0% DMSO, 8.91% MeOH).

[0777] Procedure: Stock solutions were added to the plates at each time point. Microparticle solution (360 μL / well), TA / PC (45 μL / well), and NADPH (45 μL / well) min (concentration: 1.0 μM, 0.10% DMSO, 0.891% MeOH) were added to the incubation plates. TA (15 μL / well) and microparticles (120 μL) were added to T60 and NCF60 plates. Microparticles (50 μL / well) and stop solution (150 μL / well) were added to the blank plates. At the end of each time point, 50 μL of sample was aliquoted from each well of the incubation plate. The sampling plates were shaken on a shaker for approximately 10 min, and the samples were centrifuged at 4000 rpm for 15 min. The supernatant (120 μL) was transferred for LC / MS / MS. Compounds from Table 3 were incubated with 1 μM liver microsomes (combined from multiple donors) at 37 °C in the presence of a 0.5 mg / ml microsomal protein NADPH regeneration system. Positive controls included testosterone (substrate 3A4), propafenone (2D6), and diclofenac (2C9), which were incubated with microsomes in the presence of the NADPH regeneration system. Samples were removed at time points (0 min, 5 min, 10 min, 20 min, 30 min, and 60 min) and immediately mixed with cold acetonitrile containing an internal standard (IS). Test compounds were also included that were incubated with microsomes for 60 min without the NADPH regeneration system. Individual points (n = 1) were obtained for each test condition, and samples were analyzed by LC / MS / MS. The disappearance of test compounds was assessed based on the analyte / IS peak area ratio (without a standard curve). As shown in Table 1, many compounds exhibited good stability in human and mouse liver microsomes with a T0 value exceeding 2 hours. 1 / 2 And low microparticle clearance rate.

[0778] Table 2. In vitro microsomal metabolic stability of selected compounds in CD-1 mouse and human liver microsomes.

[0779]

[0780]

[0781] Example 5. Pharmacokinetic Study of the Test Item in Male CD-1 Mice

[0782] Research Design

[0783] Table 3. Pharmacokinetic Study Design*

[0784]

[0785] *For both groups (IV and PO), plasma samples were collected at the following times: 0.083 h, 0.25 h, 0.5 h, 1.0 h, 2.0 h, 4.0 h, 8.0 h, and 24 h post-drug administration. Terminal plasma collection was performed via cardiac puncture (24 h).

[0786] **For compounds RAD001, 26, and 45, the studies were conducted in the same manner as for compound 69, with one difference: compounds 26 and 45 were administered orally (PO) at 20 mg / kg, while compounds 69 and RAD001 were administered PO at 10 mg / kg.

[0787] Animal experiments were conducted at Wuxi AppTec. Male CD1 mice aged 6–8 weeks were obtained from Hilltop Lab. Upon arrival at Wuxi AppTec, the mice were housed in an animal room with controlled environmental conditions (temperature: 20°C to 26°C; lighting: 12-hour light / dark cycle). Mice were fed a certified pellet diet (LabDiet certified rodent feed #5002). Water was provided freely. Mice were allowed to acclimatize to the facility for at least 3 days prior to the start of the experiments.

[0788] Dosage formulation

[0789] An appropriate amount of the test sample was prepared in 5% ethanol, 5% Tween 80, 5% PEG-400, and 85% water. The formulation was prepared on the day of administration and administered within 2 hours of preparation. Dosage accuracy was determined by LC-MS / MS.

[0790] Dosage administration

[0791] Administer the dosage formulation according to the facility's SOP. Determine the dosage volume individually based on the animal's body weight prior to administration.

[0792] Sample collection and plasma separation

[0793] At each predefined time point, collect approximately 40 μL of blood sample via a peripheral vein (e.g., saphenous vein). Collect the blood sample into a tube containing K2EDTA as an anticoagulant and keep it on ice until centrifugation.

[0794] Within half an hour of collection, centrifuge the blood sample at 3000g at 4°C for 5 minutes. Transfer the plasma to polypropylene tubes or 96-well plates, freeze immediately on dry ice, and store at -70±10°C until analysis by LC-MS / MS.

[0795] Bioanalytical methods and sample analysis

[0796] The concentration of the test compound in mouse plasma was determined using an LC-MS / MS method with a calibration curve having at least six non-zero calibration standards.

[0797] Data Analysis

[0798] The pharmacokinetics (PK) of the test items were analyzed using Phoenix WinNonlin software (version 8.3) and a non-compartmental analysis model. The exported PK parameters include, but are not limited to: C0, CL... p Vdss, C max T max T 1 / 2 AUC (0-t) AUC (0-inf) MRT (0-t) MRT (0-inf) And %F (bioavailability).

[0799] The results of the pharmacokinetic studies of the selected compounds were shown in the following figures: Figure 12A (Everolimus, RAD001) Figure 12B (Compound 26) Figure 12C (Compound 45) and Figure 12D In (compound 63).

[0800] Figure 12A This is a graph showing the plasma pharmacokinetic profile of RAD001 in mice after administration of IV (2 mg / kg) and PO (10 mg / kg). The Y-axis, set to a log10 scale, represents the concentration of the compound in plasma (ng / ml). The x-axis represents the time points (hours) at which plasma was collected after compound administration.

[0801] Figure 12B This is a graph showing the plasma pharmacokinetic profile of mice 26 hours after administration of IV (2 mg / kg) and PO (20 mg / kg). The Y-axis, set to a log10 scale, represents the concentration of the compound in plasma (ng / ml). The X-axis represents the time points (hours) of plasma collection after compound administration.

[0802] Figure 12C This is a graph showing the pharmacokinetic profile of compound 45 in mouse plasma after administration of IV (2 mg / kg) and PO (20 mg / kg). The Y-axis, set to a log10 scale, represents the concentration of the compound in plasma, ng / ml. The X-axis represents the time points (hours) of plasma collection after compound administration.

[0803] Figure 12DThis is a graph showing the plasma pharmacokinetic profile of mice 69 hours after administration of IV (2 mg / kg) and PO (10 mg / kg). The Y-axis, set to a log10 scale, represents the concentration of the compound in plasma, ng / ml. The X-axis represents the time points (hours) at which plasma was collected after compound administration.

[0804] Compared to RAD001, compound 69 has an improved pharmacokinetic profile, such as higher oral bioavailability (30.7% for 69 vs. 26% for RAD001), higher plasma Cmax after oral administration (12,933 ng / ml for 69 vs. 7,224 ng / ml for RAD001), and a slower plasma clearance rate (15.5 hours for 69 vs. 5.85 hours for RAD001) (comparison). Figure 12A and Figure 12D See also Table 4 below.

[0805] Table 4. Summary of pharmacokinetic studies of selected compounds in mice. Note that compounds 26 and 45 were administered orally (PO) at 20 mg / kg. Figure 12B and Figure 12C Compounds 69 and RAD001 were administered at 10 mg / kg PO ( Figure 12A and Figure 12D ).

[0806]

[0807] *IV T 1 / 2 -Ⅳ The amount of time required for the plasma concentration of the compound to decrease by 50% after administration. **Cl (ml / min / kg) -IV The clearance rate of the compound after administration. ***Vdss (L / kg) -IV The volume of distribution after administration. ****PO C max -PO administration achieves the maximum (peak) plasma concentration of the compound.

[0808] The embodiments and examples described above are intended to be illustrative and not restrictive. Those skilled in the art will recognize or be able to determine many equivalents of a particular compound, material, and procedure using only conventional experiments. All such equivalents are considered to be within the scope of and covered by the appended claims.

Claims

1. A compound of formula I: Or its pharmaceutically acceptable salt; Where R 1 It is hydrogen, C 1-6 Alkyl, heterocyclic, aryl, heteroaryl, -C 0-6 Alkylene-SO2R 4 or -C 0-6 Alkylene-SO2R 5 ; wherein C 1-6 Alkyl, heterocyclic, aryl, and heteroaryl groups are optionally surrounded by one or two R groups. 1a Group substitution; R 2 It is a heterocyclic group, aryl group, heteroaryl group, -C 0-6 Alkylene-SO2R 4 or -C 0-6 Alkylene-SO2R 5 The heterocyclic group, aryl group, and heteroaryl group are optionally surrounded by one or two R groups. 2a Group substitution; Or R 1 and R 2 Together with the attached nitrogen, it forms an optional structure with one or two R 1a N-linked heteroaryl groups or N-linked heterocyclic groups with substituent groups; R 3 Choose from the following groups: - OR a 3-6 membered heterocyclic groups and 3-6 membered heteroaryl groups; R a Choose from the following groups: H, -P(O)(R b )2、-C(O)R c -C(O)OR c C 1-6 Alkyl and C 1-6 Hydroxyalkyl; Each R b Choose independently from the following groups: H and C 1-6 alkyl; R c Choose from the following groups: H, C 1-6 Alkyl and C 1-6 Hydroxyalkyl; R 4 It is C 2-6 Alkyl, C 3-6 cycloalkyl, C 1-6 Hydroxyalkyl, heterocyclic, aryl, or heteroaryl; wherein the heterocyclic, aryl, or heteroaryl group is optionally represented by one or two R groups. 4a Group substitution; and R 5 It is a heterocyclic, aryl, or heteroaryl group; wherein the heterocyclic, aryl, or heteroaryl group is optionally surrounded by one or two R groups. 5a Group substitution; One or two R 1a R 2a R 4a and R 5a Each of the groups, when present, is independently selected from C. 1-6 Alkyl, C 1-6 Hydroxyalkyl, hydroxyl, halogen, C 1-6 Halogenated alkyl and C 1-6 alkoxy; or any two R 1a R 2a R 4a and R 5a base When groups exist on the same carbon atom, they form oxo groups together. Where R 1 When it is hydrogen, R 2 Not -C 0-6 Alkylene-SO2R 4 .

2. The compound according to claim 1, wherein the compound has formula Ia: Or its pharmaceutically acceptable salt; Where R 1 Is it hydrogen or C? 1-6 alkyl; R 2 It is optional to be controlled by one or two Rs 2a Heterocyclic, aryl, or heteroaryl groups substituted with functional groups; R 3 Choose from the following groups: - OR a 3-6 membered heterocyclic groups and 3-6 membered heteroaryl groups; R a Choose from the following groups: H, -P(O)(R b )2、-C(O)R c -C(O)OR c C 1-6 Alkyl and C 1-6 Hydroxyalkyl; Each R b Choose independently from the following groups: H and C 1-6 alkyl; R c Choose from the following groups: H, C 1-6 Alkyl and C 1-6 Hydroxyalkyl; and One or two R 2a Each of the groups, when present, is independently selected from C. 1-6 Alkyl, C 1-6 Hydroxyalkyl, hydroxyl, halogen, C 1-6 Halogenated alkyl and C 1-6 alkoxy; or any two R 2a When groups exist on the same carbon atom, they form oxo groups together.

3. The compound according to claim 1, wherein the compound has the formula Ib: Or its pharmaceutically acceptable salt; Where R 1 and R 2 Together with the attached nitrogen, it forms an optional structure with one or two R 1a N-linked heteroaryl groups or N-linked heterocyclic groups with substituent groups; R 3 Choose from the following groups: - OR a 3-6 membered heterocyclic groups and 3-6 membered heteroaryl groups; R a Choose from the following groups: H, -P(O)(R b )2、-C(O)R c -C(O)OR c C 1-6 Alkyl and C 1-6 Hydroxyalkyl; Each R b Choose independently from the following groups: H and C 1-6 alkyl; R c Choose from the following groups: H, C 1-6 Alkyl and C 1-6 Hydroxyalkyl; One or two R 1a Each of the groups, when present, is independently selected from C. 1-6 Alkyl, C 1-6 Hydroxyalkyl, hydroxyl, halogen, C 1-6 Halogenated alkyl and C 1-6 alkoxy group; or two R groups 1a When groups exist on the same carbon atom, they form oxo groups together.

4. The compound according to claim 1, wherein the compound has the formula Ic: Or its pharmaceutically acceptable salt; Where R 1 Is it hydrogen or C? 1-6 alkyl; R 2 It is -C 0-6 Alkylene-SO2R 4 or -C 0-6 Alkylene-SO2R 5 ; R 3 Choose from the following groups: - OR a 3-6 membered heterocyclic groups and 3-6 membered heteroaryl groups; R a Choose from the following groups: H, -P(O)(R b )2、-C(O)R c -C(O)OR c C 1-6 Alkyl and C 1-6 Hydroxyalkyl; Each R b Choose independently from the following groups: H and C 1-6 alkyl; R c Choose from the following groups: H, C 1-6 Alkyl and C 1-6 Hydroxyalkyl; R 4 It is C 2-6 Alkyl, C 3-6 cycloalkyl, C 1-6 Hydroxyalkyl, heterocyclic, aryl, or heteroaryl; wherein the heterocyclic, aryl, or heteroaryl group is optionally represented by one or two R groups. 4a Group substitution; R 5 It is a heterocyclic, aryl, or heteroaryl group; wherein the heterocyclic, aryl, or heteroaryl group is optionally surrounded by one or two R groups. 5a Group substitution; One or two R 4a and R 5a Each of the groups, when present, is independently selected from C. 1-6 Alkyl, C 1-6 Hydroxyalkyl, hydroxyl, halogen, C 1-6 Halogenated alkyl and C 1-6 alkoxy; or any two R 4a and R 5a When groups exist on the same carbon atom, they form oxo groups together. Where R 1 When it is hydrogen, R 2 Not -C 0-6 Alkylene-SO2R 4 .

5. The compound according to claim 1, wherein R 1 It is hydrogen and R 2 It is aryl, heteroaryl, or C 5-12 Heterocyclic groups, wherein the aryl, heteroaryl and C 5-12 The heterocyclic group is optionally surrounded by one or two R 2a Group substitution.

6. The compound according to claim 1, wherein R 1 Is it hydrogen or C? 1-6 Alkyl and R 2 It is aryl, heteroaryl, or C 5-12 Heterocyclic groups, wherein the aryl, heteroaryl and C 5-12 The heterocyclic group is optionally surrounded by one or two R 2a Group substitution.

7. The compound according to claim 1, wherein R 1 It is hydrogen and R 2 It is C 5-20 Yuanfangji, C 5-20 Monoaryl aryl or C 5-20 Heterocyclic groups, wherein the aryl, heteroaryl and C 5-12 The heterocyclic group is optionally surrounded by one or two R 2a Group substitution.

8. The compound according to claim 1, wherein R 1 Is it hydrogen or C? 1-6 Alkyl and R 2 It is C 5-20 Yuanfangji, C 5-20 Monoaryl aryl or C 5-20 Heterocyclic groups, wherein the aryl, heteroaryl and C 5-12 The heterocyclic group is optionally surrounded by one or two R 2a Group substitution.

9. The compound according to any one of claims 5-8, wherein R 2 Selected from in It is the attachment point to the rest of the compound.

10. The compound according to claim 1 or 4, wherein R 4 It is a heterocyclic, aryl, or heteroaryl group; wherein the heterocyclic, aryl, or heteroaryl group is optionally surrounded by one or two R groups. 4a Group substitution; or R 5 It is a heterocyclic, aryl, or heteroaryl group; wherein the heterocyclic, aryl, or heteroaryl group is optionally surrounded by one or two R groups. 5a Group substitution.

11. The compound according to claim 10, wherein R 4 yes in It is the attachment point to the rest of the compound.

12. The compound according to claim 10, wherein R 5 yes in It is the attachment point to the rest of the compound.

13. The compound according to any one of claims 1-12, wherein R 3 Yes - OR a And R a It is C 1-6 alkyl.

14. The compound according to claim 1, wherein the compound is selected from: Or its pharmaceutically acceptable salt.

15. The compound according to claim 14, wherein the compound is Or its pharmaceutically acceptable salt.

16. A compound, said compound being selected from the group consisting of: Or its pharmaceutically acceptable salt.

17. The compound according to any one of claims 1-16, wherein the compound is selectively bound to FKBP12.

18. The compound of claim 17, wherein, in FKBP12KO cells, the compound inhibits S6K1 phosphorylation with at least twice the efficiency of the rapamycin analog RAD001 compared to wild-type cells expressing FKBP12.

19. The compound of claim 17, wherein in FKBP12KO cells, the compound is required at a concentration at least 10 times higher than that of the rapamycin analog RAD001 to achieve 20% inhibition of S6K1 cell signaling compared to wild-type FKBP12-expressing cells.

20. The compound of claim 17, wherein, in FKBP12KO cells, the compound inhibits S6K1 phosphorylation at least ten times less efficiently than the rapamycin analog RAD001 compared to wild-type cells expressing FKBP12.

21. The compound of claim 17, wherein in FKBP12KO cells, the compound is required at a concentration at least 100 times higher than that of the rapamycin analog RAD001 to achieve 20% inhibition of S6K1 cell signaling compared to wild-type FKBP12-expressing cells.

22. The compound of claim 17, wherein in FKBP12KO cells, the compound is required at a concentration at least 10 times higher than that of the rapamycin analog RAD001 to achieve 30% inhibition of S6K1 cell signaling compared to wild-type FKBP12-expressing cells.

23. The compound of claim 17, wherein in FKBP12KO cells, the compound is required at a concentration at least 100 times higher than that of the rapamycin analog RAD001 to achieve 30% inhibition of S6K1 cell signaling compared to wild-type FKBP12-expressing cells.

24. The compound of claim 17, wherein the relative loss of potency of FKBP12 KO cells relative to wild-type cells is at least twice as great as the relative loss of potency of RAD001.

25. The compound of claim 17, wherein the relative loss of potency of FKBP12 KO cells relative to wild-type cells is at least ten times greater than the relative loss of potency of RAD001.

26. The compound of claim 17, wherein the relative loss of potency of FKBP12 KO cells relative to wild-type cells is at least 100 times greater than the relative loss of potency of RAD001.

27. A pharmaceutical composition comprising a therapeutically effective amount of the compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-26, in one or more pharmaceutically acceptable carriers.

28. A pharmaceutical combination comprising a therapeutically effective amount of any one of claims 1-26, or a pharmaceutically acceptable salt thereof, and one or more therapeutically active agents in one or more pharmaceutically acceptable carriers.

29. A method for treating a subject with a corresponding need for a disorder or disease mediated by the mTOR pathway, comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1-26 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 27 or 28.

30. A method for treating a disease or disorder in a subject, wherein a target tissue, target organ, or target cell associated with the pathology of said disease or disorder has an FKBP12 level sufficient to inhibit mTORC1, said method comprising administering to a subject in need a therapeutically effective amount of a compound according to any one of claims 1-26 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 27 or 28.

31. The method according to claim 29 or 30, wherein the compound or a pharmaceutically acceptable salt thereof has a sufficiently high affinity for FKBP12 to inhibit mTORC1 binding.

32. The method according to any one of claims 29-31, wherein the disease or disorder is selected from sarcopenia; skin atrophy; cherry angioma; seborrheic keratosis; cerebral atrophy; atherosclerosis; arteriosclerosis; emphysema; osteoporosis; osteoarthritis; hypertension; erectile dysfunction; cataract; macular degeneration; glaucoma; stroke; cerebrovascular disease (stroke); chronic kidney disease; diabetic nephropathy; impaired liver function; liver fibrosis; autoimmune hepatitis; endometrial hyperplasia; metabolic dysfunction; renal vascular disease; hearing loss; activity deficit; cognitive decline; tendon stiffness. Cardiac dysfunction such as cardiac hypertrophy and / or systolic dysfunction and / or diastolic dysfunction and / or hypertension; cardiac dysfunction leading to decreased ejection fraction; immunosenescence; Parkinson's disease; Alzheimer's disease; cancer; immunosenescence leading to cancer due to decreased immune surveillance; infections due to decreased immune function; chronic obstructive pulmonary disease (COPD); obesity; anosmia; anosmia; arthritis; epilepsy; cancer in which the tumor has elevated levels of mTORC1 signaling and / or sufficient levels of FKBP12 to allow inhibition of mTORC1; and type II diabetes.

33. A method of treating an age-related disorder or disease in a subject with corresponding needs, the method comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1-26 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 27 or 28, wherein the disorder or disease is selected from sarcopenia; skin atrophy; cherry angioma; seborrheic keratosis; cerebral atrophy; atherosclerosis; arteriosclerosis; emphysema; osteoporosis; osteoarthritis; hypertension; erectile dysfunction; cataract; macular degeneration; glaucoma; stroke; cerebrovascular disease (stroke); chronic kidney disease; diabetic nephropathy; impaired liver function; liver fibrosis; autoimmune hepatitis; Endometrial hyperplasia; metabolic dysfunction; renal vascular disease; hearing loss; activity deficit; cognitive decline; tendon stiffness; cardiac dysfunction such as cardiomegaly and / or systolic and / or diastolic dysfunction and / or hypertension; cardiac dysfunction leading to decreased ejection fraction; immunosenescence; Parkinson's disease; Alzheimer's disease; cancer; immunosenescence leading to cancer due to decreased immune surveillance; infections due to decreased immune function; chronic obstructive pulmonary disease (COPD); obesity; anosmia; anosmia; arthritis; epilepsy; cancer in which the tumor has elevated levels of mTORC1 signaling and / or sufficient levels of FKBP12 to allow inhibition of mTORC1; and type II diabetes.

34. A method of treating cancer in a subject with a corresponding need, comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1-26 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 27 or 28.

35. The method according to claim 34, wherein the cancer is selected from renal cell carcinoma, renal cell carcinoma, colorectal cancer, uterine sarcoma, endometrial cancer, breast cancer, ovarian cancer, cervical cancer, gastric cancer, fibrosarcoma, pancreatic cancer, liver cancer, melanoma, leukemia, multiple myeloma, nasopharyngeal carcinoma, prostate cancer, lung cancer, glioblastoma, bladder cancer, mesothelioma, head cancer, rhabdomyosarcoma, sarcoma, lymphoma, and cervical cancer.

36. A method of treating Alzheimer's disease or a syndrome thereof in a subject with corresponding need, comprising administering to the subject a therapeutically effective amount of the compound of any one of claims 1-26 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 27 or 28.

37. A method for inducing immune tolerance and preventing transplant rejection in subjects with appropriate need, comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1-26 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 27 or 28.

38. A method of treating graft-versus-host disease (GvHD) or a syndrome thereof in a subject with corresponding need, comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1-26 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 27 or 28.

39. A method of treating a subject with facial angiofibroma associated with tuberous sclerosis syndrome, comprising administering to the subject a therapeutically effective amount of the compound of any one of claims 1-26 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 27 or 28.

40. A method of treating an advanced, unresectable, or metastatic malignant perivascular epithelioid cell tumor in a subject with corresponding need, comprising administering to the subject a therapeutically effective amount of the compound of any one of claims 1-26 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of any one of claims 27 or 28.

41. The compound according to any one of claims 1-26 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 27 or 28, is used as a medicine.

42. The compound of any one of claims 1-26 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of any one of claims 27 or 28, for use in the treatment of disorders or diseases mediated by the mTOR pathway.

43. The compound of any one of claims 1-26 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of any one of claims 27 or 28, for use in the treatment of disorders or diseases selected from: sarcopenia; skin atrophy; cherry angioma; seborrheic keratosis; cerebral atrophy; atherosclerosis; arteriosclerosis; emphysema; osteoporosis; osteoarthritis; hypertension; erectile dysfunction; cataract; macular degeneration; glaucoma; stroke; cerebrovascular disease (stroke); chronic kidney disease; diabetic nephropathy; impaired liver function; liver fibrosis; autoimmune hepatitis; endometrial hyperplasia; metabolic dysfunction; renal vascular disease; Hearing loss; activity deficits; cognitive decline; tendon stiffness; cardiac dysfunction such as cardiomegaly and / or systolic and / or diastolic dysfunction and / or hypertension; cardiac dysfunction leading to decreased ejection fraction; immunosenescence; Parkinson's disease; Alzheimer's disease; cancer; immunosenescence leading to cancer due to decreased immune surveillance; infections due to decreased immune function; chronic obstructive pulmonary disease (COPD); obesity; loss of taste; loss of smell; arthritis; epilepsy; cancer in which the tumor has elevated levels of mTORC1 signaling and / or sufficient levels of FKBP12 to suppress mTORC1; and type II diabetes.

44. The compound according to any one of claims 41-43, wherein the compound is selected from... Or a pharmaceutically acceptable salt.

45. The compound according to any one of claims 41-44, wherein the compound is selected from... Or its pharmaceutically acceptable salt.

46. ​​A combination therapy comprising a therapeutically effective amount of the compound of any one of claims 1-26 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of claim 27 or 28, and an anti-CD40 antibody.

47. The combination therapy of claim 46, for use in the treatment and / or prevention of Alzheimer's disease, GvHD, facial angiofibroma associated with tuberous sclerosis syndrome, advanced unresectable or metastatic malignant perivascular epithelioid tumors, and transplant rejection.

48. The combination therapy of claim 46, for use in the treatment of human diseases selected from the group consisting of cancer, diabetes, obesity, neurological disorders, genetic disorders and other age- or aging-related diseases.

49. A method of treating Alzheimer's disease or syndrome in a subject with corresponding need, comprising administering to the subject a therapeutically effective amount of the combination therapy according to claim 46.

50. A method for inducing immune tolerance and preventing transplant rejection in subjects with appropriate need, comprising administering to the subject a therapeutically effective amount of the combination therapy according to claim 46.

51. A method of treating graft-versus-host disease (GvHD) or syndrome in a subject with corresponding need, comprising administering to the subject a therapeutically effective amount of the combination therapy according to claim 46.

52. A method of treating a subject with facial angiofibroma associated with tuberous sclerosis syndrome, comprising administering to the subject a therapeutically effective amount of the combination therapy according to claim 46.

53. A method of treating an advanced, unresectable, or metastatic malignant perivascular epithelioid tumor in a subject with corresponding need, comprising administering to the subject a therapeutically effective amount of the combination therapy according to claim 46.

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