RU2807188C2 - Rapamycin derivatives - Google Patents

Abstract

FIELD: rapamycin derivatives.

SUBSTANCE: invention is related to a compound of formula (I) or a pharmaceutically acceptable salt thereof, where R₁ is selected from the group consisting of hydroxy, , , and ; and R₂ is selected from the group consisting of , , , where m equals 0; n is 1 or 2; and R₃ represents hydrogen, C_1-6 alkyl or hydroxy-C_1-6 alkyl. The invention is also related to a pharmaceutical composition for inhibiting mTOR activity based on the specified compound.

EFFECT: new compounds and a pharmaceutical composition based on them have been obtained, which can be used in medicine to prevent or treat a disorder or disease mediated by the mTOR pathway.

33 cl, 16 dwg, 20 ex

Description

PRIORITY STATEMENT

This application claims benefit from U.S. Patent Application Serial No. 62/563312, filed September 26, 2017, which is incorporated herein by reference in its entirety.

FIELD OF TECHNOLOGY TO WHICH THE INVENTION RELATES

The present invention provides 32-deoxyrapamycin derivatives and relates to methods of using them.

BACKGROUND OF THE ART

In mammalian cells, the kinase target of rapamycin (mTOR) exists in two distinct multiprotein complexes described as the mTORC1 complex and the mTORC2 complex, both of which respond to nutrient and energy availability and mediate the integration of incoming signals from growth factors and signal transduction in the event of stress. mTORC1 integrates signals from growth factors and nutrients and controls cell growth and metabolism. See Laplante M. et al. Cell. (2012) 149(2):274-93. mTORC1 is a key regulator of protein translation and autophagy. The mTORC1 complex is sensitive to allosteric inhibitors of mTOR, such as rapamycin and rapamycin analogs (so-called “rapalogues”). The mechanism of action of rapamycin and previously prepared rapalogues involves the formation of an intracellular complex with FK506-binding proteins, one of FKBP12, FKBP51 or FKBP52 (these three FKBP species will be referred to herein as “FKBP” or “FKBP species”), followed by binding of the FKBP complex -rapalog with FRB domain (FK506-rapamycin binding) mTOR. Cm. et al. Mol Cell Biol. (2013) 33(7):1357–1367. "Large FK506-Binding Proteins Shape the Pharmacology of Rapamycin". This interaction of the FKBP-rapalog complex with mTORC1 leads to allosteric inhibition of the complex. Rapamycin and rapalogues such as RAD001 (everolimus; Afinitor®) have gained clinical relevance by inhibiting the activity of mTORC1, which is associated with disorders associated with the proliferation of both benign and malignant cells. See Royce ME et al. Breast Cancer (Auckl). (2015) 9:73–79; Pleniceanu O. et al. Kidney Int Rep. (2018) 3(1):155-159.

Rapamycin is a well-known macrolide antibiotic produced byStreptomyces hygoscopius, cm. for example, McAlpine, J.B.,et al.J Antibiotics (1991) 44:688; Schreiber, S. L.;et al., J. Am. Chem. Soc. (1991) 113:7433; US Patent No. 3,929,992 The following atom numbering convention for rapamycin and its derivatives as used herein is shown below.

Rapamycin is an effective immunosuppressant and has also been shown to have antitumor and antifungal activity. It has been shown to be useful in the prevention or treatment of systemic lupus erythematosus, pneumonia, insulin-dependent diabetes mellitus, skin disorders such as psoriasis, smooth muscle cell proliferation and intimal thickening after vascular injury, T-cell leukemia/lymphoma in adults, malignant carcinomas, inflammatory heart disease, anemia and increased axonal sprouting. However, its applicability as a pharmaceutical agent is limited by its extremely low and variable bioavailability. Moreover, rapamycin is difficult to formulate, making it difficult to obtain stable medicinal compositions.

To solve these problems, numerous rapalogues have been (semi)synthesized. Water-soluble prodrugs were prepared by derivatizing rapamycin at C28 and 40 to form glycinate, propionate, and pyrrolidinobutyrate prodrugs (US Pat. No. 4,650,803). Other rapamycin analogs include monoacyl and diacyl analogs (US Pat. No. 4,316,885), acetal analogs (US Pat. No. 5,151,413), silyl ethers (U.S. Pat. No. 5,120,842), hydroxy esters (U.S. Pat. No. 5,362,718), and aryl, alkyl, alkenyl, and alkynyl analogs. analogs (US patents No. 5665772; 5258389; 6384046; WO 97/35575). Modifications of rapamycin include demethylation, removal or substitution of one or more methoxy groups; removal, derivatization or replacement of one or more hydroxy moieties; reducing, removing or derivatizing one or more of the ketone moieties; replacement of a 6-membered pipecolate ring with a 5-membered prolyl ring; alternatively replacing a cyclohexyl ring with a substituted cyclopentyl ring. Illustrative examples of patent literature in the art are US Patent No. 5527907, WO 96/41865 and WO 99/36553, which describe a wide variety of rapalogues aimed at eliminating the immunosuppressive side effects of rapamycin. US Pat. No. 5,985,890 discloses examples of 32-deoxorapamycin analogues, including 32-deoxorapamycin itself. Such compounds have been reported to have an improved pharmacological profile compared to rapamycin and better stability.

The rapalogues described in the literature above have been disclosed as useful for treating the same disorders as rapamycin. US Patent No. 8906374 and US Patent No. 9669032 disclose use in cancer. US Pat. No. 9,358,236 discloses use in neurodegenerative disorders.

In animal models, rapalogs increase lifespan and delay the onset of age-related diseases. Aging, like other biological processes, is regulated by signaling pathways such as the TOR pathway (referred to as "TOR" in this case to include systems involving yeast and C elegans , in which the mammalian equivalent protein [mTOR] is simply called "TOR""]) and, in mammals, the mTORC1 pathway. Modulation 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, inhibition of the TOR pathway by genetic mutation increased lifespan in yeast, C elegans , and Drosophila, and inhibition of the mTORC1 pathway increased lifespan in 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; Vellai et al. , Nature (2003) 426:620). In addition, the mTORC1 inhibitor rapamycin increased lifespan in mice even when administered late in life (Harrison et al. , Nature (2009) 460(7253):392-395). Such data raise the possibility that drugs targeting the mammalian TOR (mTOR) pathway will have therapeutic effects in aging and age-related diseases in humans. For example, WO 2008/022256 describes topical methods and formulations containing an mTOR inhibitor for the treatment or prevention of an age-related disease. A report of a clinical trial using rapamycin in older men was described by M. Leslie in Science, 2013, 342. J. Mannick et al. describes in Sci Transl Med. (2014) 6(268): 268ra179 that mTOR inhibition improves immune function in the elderly. However, researchers have been cautious in considering the use of currently available mTOR inhibitors in human aging trials due to their side effects (including immunosuppression, cytopenias, stomatitis, gastrointestinal distress, and interstitial pneumonitis).

Studies of focal cortical dysplasia (FCD) and tuberous sclerosis complex (TSC) in animals and humans have implicated the mTOR pathway in mediating cellular and molecular changes leading to the formation of cortical pathologies and the manifestation of epilepsy (Wong et al. , Experimental Neurology (2013) 244: 22-26 Focal cortical dysplasia (FCD) is a cortical developmental disorder that is the most common cause of refractory epilepsy in children and the second/third most common cause of drug-resistant seizures in adults (Kabat J, et al. , Pol J Radiology (2012) 77(2) 35-43) The effect of mutations in the tuberous sclerosis complex (TSC), including tuberous sclerosis complex-1 (TSC1) and tuberous sclerosis complex-2 (TSC2), is higher in along the mTOR pathway, which leads to widespread development of benign tumors, mental retardation, and a high incidence of epilepsy (Manning et al. , Identification of the tuberous sclerosis complex-2 tumor suppressor gene product tuberin as a target of the phosphoinositide 3-kinase/akt pathway, Mol. Cell, (2002) 10: 151-162; Inoki et al. , Dysregulation of the TSC-mTOR pathway in human disease, Nat. Genet, (2005), 37:19-24; and Holmes and Stafstrom, Tuberous sclerosis complex and epilepsy: recent developments and future Challenges, Epilepsia, (2007) 48:617-630).

Abnormal activation of mTOR interferes with normal brain development and leads to epilepsy. Treatment with rapamycin, which inhibits the mTORC1 pathway, has been shown to reduce structural abnormalities and reduce seizure intensity in TSC and PTEN mouse models (Ehninger et al. , Reversal of learning deficits in a Tsc2+/- mouse model of tuberous sclerosis; Nat. Med., (2008), 843-848; Meikle et al. , Response of a neuronal model of tuberous sclerosis to mammalian target of rapamycin, mTOR inhibitors: effects on mTORC1 and Akt signaling leading to improved survival and function; J. Neuroscience. , (2008) 28:5422-5432; Zeng et al. , Rapamycin prevents epilepsy in a mouse model of tuberous sclerosis complex; Ann. Neurol., (2008) 63:444-453; Ljungberg et al. , Rapamycin suppresses seizures and neuronal hypertrophy in a mouse model of cortical dysplasia; (2009) pp. 389-398; and Zhou et al. , Pharmacological inhibition of mTORC1 suppresses anatomical, cellular, and behavioral abnormalities in neural-specific Pten knock-out mice, J. Neurosci ., (2009), 29:1773-1783). In addition, pharmacological inhibition of the mTOR pathway either before or immediately after neurological injury can prevent pathological changes in the animal brain and the development of spontaneous recurrent epileptic seizures in a model of acquired epilepsy (Zeng et al. , The mammalian target of rapamycin signaling pathway mediates epileptogenesis in a model of temporal lobe epilepsy; J. Neurosci., (2009) pp. 6964-6972). Therefore, rapamycin and rapalogues are also considered to have potential value in these indications.

Mitochondrial myopathy (MM) is the most common manifestation of acquired mitochondrial disease and exhibits a multifaceted tissue-specific stress response: (1) a transcriptional response involving the metabolic cytokines FGF21 and GDF15; (2) remodeling of one-carbon metabolism; and (3) the reaction of unfolded mitochondrial proteins. In Cell Metabolism 26, 419–428, August 1, 2017. Khan et al. describe such processes as part of a single integrated mitochondrial stress response (ISRmt) that is controlled by mTORC1 in skeletal muscle. The mtDNA replication defect activates mTORC1, which triggers the integrated mitochondrial stress response by activating ATF4, inducing de novo nucleotide and serine synthesis, the 1C cycle, and producing FGF21 and GDF15. Inhibition of mTORC1 with rapamycin reduced the expression of all components of ISRmt (integrated mitochondrial stress response), improved all MM parameters, and reversed MM progression even at a late stage, without inducing mitochondrial biogenesis. Therefore, rapamycin and rapalogues are also considered to have potential value in these indications.

There remains a need to provide new mTOR inhibitors that are good drug candidates. In particular, preferred compounds should have at least the ability to inhibit mTORC1 and be readily absorbed from the gastrointestinal tract, be sufficiently metabolically stable, and have advantageous pharmacokinetic properties. In addition, an ideal drug candidate will be able to exist in a physical form that is stable, non-hygroscopic, and suitable for formulation.

SHORT DESCRIPTION

The compounds of structural formula (I) are mTORC1 inhibitors and are therefore potentially useful in the treatment of a wide variety of disorders, in particular age-related disorders or diseases and disorders for which treatment with rapalogues is currently approved. Complete reduction of the ketone at C32 and replacement of the methoxy group at C16 with a cyclic N-containing aliphatic ring system such as a cyclic amine, amide or sultam provides compounds with the above desired benefits that demonstrate a balance between proper activity, stability and bioavailability.

In one aspect, the present invention provides compounds of structural formula (I):

or a pharmaceutically acceptable salt thereof, wherein

R ₁ is selected from the group consisting of hydroxy, And

R ₂ is selected from the group consisting of

, Where

m is 0, 1, 2 or 3;

n is 1, 2 or 3;

o equals 1, 2, 3, 4, 5 or 6;

p equals 1, 2, 3, 4 or 5;

q is equal to 1, 2, 3, 4 or 5, with the sum of p and q being 2, 3, 4, 5 or 6;

r equals 2, 3 or 4;

s is equal to 2, 3 or 4, with the sum of r and s being 4, 5 or 6;

X is O, S, NR ₆ or SO ₂ ;

R ₃ represents hydrogen, C _1-6 alkyl, hydroxy-C _1-6 alkyl, C _3-8 cycloalkyl-C _0-6 alkyl or phenyl-C _0-6 alkyl;

R ₄ represents hydrogen;

R ₅ is hydrogen, hydroxy or cyano; or R ₄ and R ₅ together form =O; And

R ₆ is hydrogen, C _1-6 alkyl, C _3-8 cycloalkyl-C _0-6 alkyl, phenyl-C _0-6 alkyl, C _1-6 alkyl-CO-, C _3-8 cycloalkyl-C _{0- 6} alkyl-CO-, C _1-6 alkyl-SO ₂ - or C _3-8 cycloalkyl-C _0-6 alkyl-SO ₂ -.

In one embodiment, the present invention provides a pharmaceutical composition containing a therapeutically effective amount of a compound of structural formula (I) or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers.

In one embodiment, the present invention provides a pharmaceutical combination comprising a therapeutically effective amount of a compound of structural formula (I) or a pharmaceutically acceptable salt thereof and one or more therapeutically active agents.

In another aspect, the present invention provides a method of treating a disorder or disease mediated by the mTOR pathway in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of structural formula (I), or a pharmaceutical composition, or a pharmaceutical combination thereof . In another aspect, the present invention provides a method of treating a disease or disorder in a subject, wherein the target tissue or organ associated with pathology associated with the disease or disorder has levels of FKBP12 sufficient to inhibit mTORC1, the method comprising administering to the subject in need in this, a therapeutically effective amount of a compound of structural formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition, or a pharmaceutical combination thereof.

In one embodiment, the target, which is a tissue or organ associated with a pathology caused by a disease or disorder to be treated with a compound of structural formula (I), characterized by levels of FKBP12 sufficient to inhibit mTORC1, is determined empirically, for example, using an inhibitor specific for FKBP12 versus rapamycin or RAD001.

In another aspect, the present invention provides a method of treating a disease or disorder in a subject having or previously determined to have levels of FKBP12 sufficient to inhibit mTORC1, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of structural formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition, or a pharmaceutical combination based thereon.

In one embodiment, the subject is characterized or previously determined to have levels of FKBP12 in the target tissue, organ, or cells sufficient to inhibit mTORC1.

In another aspect, the present invention provides a method of treating an age-related disease or disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of structural formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition, or a combination thereof ,

In one embodiment, the disease or disorder is selected from sarcopenia, skin atrophy, types of senile hemangioma, types of seborrheic keratosis, cerebral atrophy, also called dementia, atherosclerosis, arteriosclerosis, emphysema, osteoporosis, osteoarthritis, high blood pressure, erectile dysfunction, types of cataracts , macular degeneration, glaucoma, stroke, cerebrovascular disease(s), chronic kidney disease, diabetes-associated kidney disease, liver dysfunction, liver fibrosis, autoimmune hepatitis, endometrial hyperplasia, metabolic disorder, renovascular disease, hearing loss, impaired ability to movement (for example, frailty), decreased cognitive ability, tendon stiffness, cardiac dysfunction such as cardiac hypertrophy and/or systolic and/or diastolic dysfunction and/or hypertension, cardiac dysfunction that leads to decreased ejection fraction, aging of the immune system, Parkinson's disease, Alzheimer's disease, cancer, aging of the immune system 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 associated with diabetes, such as kidney failure, blindness and neuropathy.

In another aspect, the present invention provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of structural formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition, or a pharmaceutical combination thereof , where the disorder or disease is selected from

- acute or chronic organ or tissue transplant rejection;

- graft-related vasculopathies;

- proliferation and migration of smooth muscle cells, leading to thickening of the vascular intima, blockage of blood vessels, obstructive coronary atherosclerosis, restenosis;

- autoimmune diseases and inflammatory conditions;

- diseases associated with the treatment and prevention of asthma;

- diseases associated with multiple drug resistance (MDR);

- infections caused by fungi;

- inflammation;

- infections;

- age-related diseases;

- neurodegenerative diseases;

- proliferative disorders, in particular cancer;

- epileptic seizures and disorders associated with epileptic seizures and

- mitochondrial myopathy and mitochondrial stress.

In another aspect, the present invention provides a method of treating cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of structural formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition, or a pharmaceutical combination thereof.

In one embodiment, the method further comprises the use of a PD-1/PDL-1 inhibitor.

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

In one embodiment, the disorder is a liver disorder that involves a process of fibrosis and/or inflammation, for example, liver fibrosis, which occurs during end-stage liver disease; cirrhosis of the liver; liver failure caused by toxicity; non-alcoholic steatohepatitis or NASH and alcoholic steatosis.

In one embodiment, the disorder is a kidney disorder that involves a process of fibrosis or inflammation in the kidney, for example, renal fibrosis, which occurs as a result of acute kidney injury, leading to chronic kidney disease and diabetic nephropathy.

In one embodiment, the disorder is cardiac dysfunction, for example myocardial infarction or cardiac hypertrophy. In one embodiment, the cardiac dysfunction is systolic and/or diastolic dysfunction. In one embodiment, the cardiac dysfunction is hypertension. In one embodiment, cardiac dysfunction results in decreased ejection fraction.

In one embodiment, the disorder is an aging immune system leading to cancer due to decreased immune surveillance.

In one embodiment, the disorder is a cancer, including tumors that are treated with immunotherapy and tumors that have previously been treated with either rapamycin, everolimus, or another rapalogue. In one embodiment, cancer includes tumors that have been shown to have the mTOR pathway activated, including conditions in which a mutation in the Tsc1 gene occurs, or where the tumor microenvironment is appropriately treated with rapalog.

Details of one or more embodiments of the present invention are set forth herein. Other features, objects and advantages of the present invention will be apparent from the drawings, detailed description, examples and claims.

BRIEF DESCRIPTION OF GRAPHIC MATERIALS

Figure 1A is an image of the cocrystal structure of Example 1 with FKBP12. The absolute configuration along C16 is ( S ).

Figure 1B shows the structure of Example 1 with the FKBP12 protein removed.

Figure 2A is an image of the cocrystal structure of Example 2 with FKBP12. The absolute configuration along C16 is ( R ).

Figure 2B shows the structure of Example 2 with the FKBP12 protein removed.

Figure 3 is a line graph showing the pharmacokinetic profile example 2 in rats after administration of a single p.o. dose of 3 mg/kg. The Y-axis shows the blood concentrations of Example 2 (nM). The x-axis shows the time (in hours) of blood collection after administration of Example 2. Data are means ± standard deviation for 3 rats.

Figure 4A is a bar graph showing comparative blood concentrations of RAD001 (white bars) and Example 2 (dark bars) in rats receiving single oral doses of 3, 10 and 30 mg/kg of either compound. Compound concentrations were measured at 3 and 24 hours (h) post-dose. Data represent means ± standard deviation of 5-6 rats in each group. Rats that were below the limit of quantitation were excluded from data analysis: this applies to the RAD001-treated groups. An asterisk (*) indicates a significant difference between the respective groups treated with RAD001 and example 2. ** P < 0.01, *** P <0.001;****P<0.0001,Student's t test. BQL means below the limit of quantitation. The y-axis shows the blood concentrations of the compound (nM). The x-axis shows the time (3 and 24 hours) after the oral dose and the doses administered orally (3, 10, and 30 mg/kg).

Figure 4B is a bar graph showing comparative brain concentrations of RAD001 (white bars) and Example 2 (dark bars) in rats receiving single oral doses of 3, 10 and 30 mg/kg of either compound. Compound concentrations were measured at 3 and 24 hours (h) post-dose. Data are means ± standard deviation. Each group used 5-6 rats. Rats that were below the limit of quantitation were excluded from data analysis: this applies to the RAD001-treated groups. An asterisk (*) indicates a significant difference between the respective groups treated with RAD001 and example 2. ** P < 0.01, *** P <0.001;****P<0.0001,Student's t test. BQL means below the limit of quantitation. The y-axis shows the brain concentrations of the compound (nM). The x-axis shows time (3 and 24 hours) post-dose and orally administered doses (3, 10, and 30 mg/kg).

Onfigure 4Cshown concentration of example 2 in the blood of rats after intravenous (i.v.) and oral (p.o.) dosing. The Y-axis shows the blood concentrations of Example 2 (nM). The x-axis shows the time (in hours) of blood collection after administration of Example 2. Data are means ± standard deviation for 3 rats.

On4D figureshown concentration of RAD001 in the blood of rats after intravenous (i.v.) and oral (p.o.) dosing. The Y-axis shows blood concentrations of RAD001 (nM). The x-axis shows the time (in hours) of blood collection after RAD001 administration. Data represent means ± standard deviations of 3 rats.

Figures 5A - 5D show that Example 2 provides inhibition of the mTORC1 pathway in rat liver. Rats were administered a single oral dose of Example 2 at 3, or 10, or 30 mg/kg and liver samples were collected at 3 hours and 24 hours after dosing. Rats treated with vehicle (Veh) were used as controls. Figures ( 5A ) and ( 5C ) show immunoblotting images of phosphorylated (p-) and total (t-) S6 proteins from rat liver samples treated with vehicle or Example 2 at 3 or 10 or 30 mg/day. kg and analyzed 3 hours ( 5A ) and 24 hours ( 5C ) after treatment. The histograms in Figures ( 5B ) and ( 5D ) demonstrate densitometric quantification of the ratio of p-S6 to t-S6 at 3 hours ( 5B ) and 24 hours ( 5D ) after treatment. In the histograms ( 5B and 5D ), random averages that indicate p-S6/t-S6 ratios are shown above each bar. X-axes represent doses administered orally (3, 10, 30 mg/kg). Y axes represent arbitrary units. Six rats were used in each experimental group. Data are means ± standard deviation. Data were analyzed using one-way ANOVA followed by Dunnett's multiple comparison tests, where the mean values from all groups were compared with the values for the vehicle-treated group. ** P < 0.01, **** P < 0.001, ns - not significant.

Figures 6A to 6C show inhibition of S6K1(Thr389) in wild-type 293T cells ( 6A ), FKBP12 knockdown cells ( 6B ), and FKBP12 knockout cells ( 6C ) following treatment with RAD001 (dashed line) and Example 2 (solid line) ). Cells were treated in triplicate. The Y axis represents the percentage of inhibition relative to the level of S6K1(Thr389) in cells treated with medium plus DMSO. The x-axis represents the concentrations of RAD001 and Example 2.

DETAILED DESCRIPTION OF THE INVENTION

In a first aspect, the present invention provides a compound of formula (I),

or a pharmaceutically acceptable salt thereof, wherein

R ₁ is selected from the group consisting of hydroxy, And

R ₂ is selected from the group consisting of

, Where

m is 0, 1, 2 or 3;

n is 1, 2 or 3;

o equals 1, 2, 3, 4, 5 or 6;

p equals 1, 2, 3, 4 or 5;

q is equal to 1, 2, 3, 4 or 5, with the sum of p and q being 2, 3, 4, 5 or 6;

r equals 2, 3 or 4;

s is equal to 2, 3 or 4, with the sum of r and s being 4, 5 or 6;

X is O, S, NR ₆ or SO ₂ ;

R ₃ represents hydrogen, C _1-6 alkyl, hydroxy-C _1-6 alkyl, C _3-8 cycloalkyl-C _0-6 alkyl or phenyl-C _0-6 alkyl;

R ₄ represents hydrogen;

R ₅ is hydrogen, hydroxy or cyano; or R ₄ and R ₅ together form =O; And

R ₆ is hydrogen, C _1-6 alkyl, C _3-8 cycloalkyl-C _0-6 alkyl, phenyl-C _0-6 alkyl, C _1-6 alkyl-CO-, C _3-8 cycloalkyl-C _{0- 6} alkyl-CO-, C _1-6 alkyl-SO ₂ - or C _3-8 cycloalkyl-C _0-6 alkyl-SO ₂ -.

In one embodiment, the present invention provides a compound of formula (I),

or a pharmaceutically acceptable salt thereof, wherein

R ₁ is selected from the group consisting of hydroxy, And

R ₂ is selected from the group consisting of

, Where

m is 0, 1, 2 or 3;

n is 1, 2 or 3;

o equals 1, 2, 3, 4, 5 or 6;

p equals 1, 2, 3, 4 or 5;

q is equal to 1, 2, 3, 4 or 5, with the sum of p and q being 2, 3, 4, 5 or 6;

r equals 2, 3 or 4;

s is equal to 2, 3 or 4, with the sum of r and s being 4, 5 or 6;

X is O, S, NR ₆ or SO ₂ ;

R ₃ represents hydrogen, C _1-6 alkyl, C _3-8 cycloalkyl-C _0-6 alkyl or phenyl-C _0-6 alkyl;

R ₄ represents hydrogen;

R ₅ is hydrogen, hydroxy or cyano; or R ₄ and R ₅ together form =O; And

R ₆ is hydrogen, C _1-6 alkyl, C _3-8 cycloalkyl-C _0-6 alkyl, phenyl-C _0-6 alkyl, C _1-6 alkyl-CO-, C _3-8 cycloalkyl-C _{0- 6} alkyl-CO-, C _1-6 alkyl-SO ₂ - or C _3-8 cycloalkyl-C _0-6 alkyl-SO ₂ -.

Definitions

Unless otherwise indicated, the term "compounds of the present invention" or "compound of the present invention" refers to the compounds of formula (I) and illustrative compounds and their salts, as well as all stereoisomers (including diastereoisomers and enantiomers), rotamers, tautomers and isotope-labeled compounds (including deuterium substitutions), as well as originally formed fragments.

The designation used in this document represents part of the variable associated with the main molecule and includes both (R)- and (S)-stereochemistry. For example, if ^R2 is , is the R ₂ moiety bound to C16 and includes both (R)- and (S)-stereochemistry.

As used in this document, the term "C_1-6alkyl" means a straight or branched chain hydrocarbon radical consisting only of carbon and hydrogen atoms, free of unsaturation, containing from one to six carbon atoms, and which is attached to the rest of the molecule by a single bond. The term "C"_1-4alkyl" should be interpreted accordingly. Examples C_1-6alkyl include, but are not limited to, methyl, ethyl,n-propyl, 1-methylethyl (isopropyl),n-butyl,n-pentyl and 1,1-dimethylethyl (rubs-butyl).

As used herein, the term “hydroxy-C _1-6 alkyl” refers to an alkyl group substituted with one or more -OH groups. Examples of hydroxy-C _1-6 alkyl groups include HO-CH ₂ -, HO-CH ₂ CH ₂ - and -CH ₂ -CH(OH)-.

As used herein, the term "C _3-8 cycloalkyl-C _0-6 alkyl" refers to a stable monocyclic saturated hydrocarbon radical consisting solely of carbon and hydrogen atoms, containing from three to eight carbon atoms, and which is attached to the rest of the molecule by single bond or via a C _1-6 alkyl radical as defined above. Examples of C _3-8 cycloalkyl-C _0-6 alkyl include, but are not limited to, cyclopropyl, cyclopropylmethyl, cyclobutyl, cyclobutylethyl, cyclopentyl, cyclopentylpropyl, cyclohexyl, cycloheptyl, and cyclooctyl.

As used herein, the term “phenyl-C _0-6 alkyl” refers to a phenyl ring attached to the remainder of the molecule by a single bond or by a C _1-6 alkyl radical as defined above. Examples of phenyl- _C0-6 alkyl include, but are not limited to, phenyl and benzyl.

As used herein, the term “hydroxy” or “hydroxyl” refers to -OH.

Various (listed) embodiments of the present invention are described herein. It should be understood that the features specified in each embodiment can be combined with other specified features to obtain additional embodiments of the present invention.

Embodiment 1. The compound of formula (I) or a pharmaceutically acceptable salt thereof as described above.

Embodiment 2. The compound of Embodiment 1, or a pharmaceutically acceptable salt thereof, wherein R ₁ is hydroxy.

Embodiment 3. The compound of Embodiment 1 or 2, or a pharmaceutically acceptable salt thereof, wherein R ₂ is selected from , in particular , where m, n, X and R ₃ are defined above.

Embodiment 4. The compound according to any of embodiments 1-3, or a pharmaceutically acceptable salt thereof, wherein R ₂ is , and n is 1, 2 or 3. In one embodiment, R ₂ is .

Embodiment 5. The compound according to any of embodiments 1-4, or a pharmaceutically acceptable salt thereof, wherein R ₂ is .

Embodiment 6. The compound according to any of embodiments 1-4, or a pharmaceutically acceptable salt thereof, wherein R ₂ is .

Embodiment 7. The compound according to any of embodiments 1-4, or a pharmaceutically acceptable salt thereof, wherein R ₂ is . In one embodiment, R ₃ is hydrogen, C _1-6 alkyl, or hydroxy-C _1-6 alkyl.

Embodiment 8. The compound according to any one of embodiments 1 to 5 of formula (I)-A,

or a pharmaceutically acceptable salt thereof, wherein R ₁ is selected from the group consisting of hydroxy, . In one embodiment, R ₁ represents hydroxyl. In one embodiment, R ₂ is as defined in formula (I). In one embodiment, the C16 position has (R)-stereochemistry. In one embodiment, the C16 position has (S)-stereochemistry.

Embodiment 9. The compound of any one of Embodiments 1 to 8, or a pharmaceutically acceptable salt thereof, wherein said compound is C16-(1,1-dioxidoisothiazolidin-2-yl)-C32-deoxyrapamycin (Compound 1):

Embodiment 10. The compound of any one of Embodiments 1-9, or a pharmaceutically acceptable salt thereof, present as a single C16 diastereoisomer. In one embodiment, the C16 position has (R)-stereochemistry. In one embodiment, the C16 position has (S)-stereochemistry.

Embodiment 11. The compound of any one of Embodiments 1-9, or a pharmaceutically acceptable salt thereof, provided as a C16 diastereomeric mixture.

Embodiment 12. A compound according to any one of embodiments 1 to 10 of formula (I)-B,

or a pharmaceutically acceptable salt thereof, wherein R ₁ and R ₂ are as defined in formula (I). In one embodiment, R ₂ is

Embodiment 13. A compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein said compound is (S)-C16-(1,1-dioxidoisothiazolidin-2-yl)-C32-deoxorapamycin ( Example 1 ):

Embodiment 14. A compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein said compound is (R)-C16-(1,1-dioxidoisothiazolidin-2-yl)-C32-deoxorapamycin ( Example 2 ):

Embodiment 15. A compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein said compound is selected from the following.

Compound Structure Example 3
* Absolute stereochemical configuration for C16 is not specified Example 4
* Absolute stereochemical configuration for C16 is not specified Example 5
* Absolute stereochemical configuration for C16 is not specified Example 6
Example 7
diastereomers at C16
* Absolute stereochemical configuration for C16 is not specified Example 8
Example 9
diastereomers at C16
* Absolute stereochemical configuration for C16 is not specified Example 10
Example 11
diastereomers at C16
* Absolute stereochemical configuration for C16 is not specified Example 12
Example 13
Separate diastereomer at C16
* Absolute stereochemical configuration for C16 is not specified Example 14
Example 15
diastereomers at C16
* Absolute stereochemical configuration for C16 is not specified Example 16 Example 17
* Absolute stereochemical configuration for C16 is not specified Example 18
* Absolute stereochemical configuration for C16 is not specified Example 19
* Absolute stereochemical configuration for C16 is not specified

Definitions of specific functional groups and chemical terms are described in more detail below. Chemical elements are identified according to the Periodic Table of the Elements, CAS version,Handbook of Chemistry and Physics, 75th ed., inside the cover, and specific functional groups are usually defined as described therein. Additionally, general principles of organic chemistry, as well as specific functional moieties and reactivity, are described in Thomas Sorrell,Organic Chemistry, University Science Books, Sausalito, 1999; Smith and MarchMarch's Advanced Organic Chemistry, 5^th Edition, John Wiley & Sons, Inc., New York, 2001; Larock,Comprehensive Organic Transformations, VCH Publishers, Inc., New York, 1989 and Carruthers,Some Modern Methods of Organic Synthesis, 3^rd Edition, Cambridge University Press, Cambridge, 1987.

Depending on the choice of starting materials and procedures, the compounds of the present invention may be in the form of one of the possible stereoisomers or in the form of mixtures thereof in the case of stereocenters not limited to formula (I), formula (I)-A and formula (I)-B, for example, as pure optical isomers or as mixtures of stereoisomers, such as racemates and mixtures of diastereoisomers, depending on the number of asymmetric carbon atoms. The present invention is intended to include all such possible stereoisomers, including racemic mixtures, mixtures of diastereoisomers, and optically pure forms. Optically active ( R )- and ( S )-stereoisomers can be prepared using chiral synthons or chiral reagents or isolated using traditional techniques. If the compound contains a double bond, the substituent may have the E- or Z-configuration. If the compound contains a disubstituted cycloalkyl, the cycloalkyl substituent may have a cis or trans configuration. It is also intended to include all tautomeric forms.

The term "tautomers" refers to compounds that are interchangeable forms of a particular compound structure and that differ in the movement of hydrogen atoms and electrons. Thus, two structures can be in equilibrium due to the movement of π electrons and an atom (usually H). For example, enols and ketones are tautomers because they rapidly interconvert when treated with either an acid or a base. Another example of tautomerism is the acy and nitro forms of phenylnitromethane, which are similarly formed upon treatment with an acid or base. Tautomeric forms may be relevant to achieving optimal chemical reactivity and biological activity of the compound of interest.

As used herein, the terms “salt” or “salts” mean the acid addition or base addition salt of a compound of the present invention. "Salts" include, in particular, "pharmaceutically acceptable salts". The term "pharmaceutically acceptable salts" means salts that retain the biological effectiveness and properties of the compounds of the present invention and that are generally not biologically or otherwise undesirable. In many cases, the compounds of the present invention are capable of forming acidic and/or basic salts due to the presence of amino and/or carboxyl groups or the like. Pharmaceutically acceptable salts are well known in the art. For example, Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences (1977) 66:1-19, incorporated herein by reference.

Pharmaceutically acceptable acid addition salts can be formed using inorganic acids and organic acids.

Inorganic acids from which salts can be prepared include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like.

Organic acids from which salts can be prepared include, for example, acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, sulfosalicylic acid, etc.

Pharmaceutically acceptable base addition salts can be formed with inorganic and organic bases.

Inorganic bases from which salts can be prepared include, for example, ammonium and metal salts from Groups I to XII of the periodic table of elements. In certain embodiments, the salts are derived from sodium, potassium, ammonium, calcium, magnesium, iron, silver, zinc, and copper; in particular, suitable salts include ammonium, potassium, sodium, calcium and magnesium salts.

Organic bases from which salts can be derived include, for example, primary, secondary and tertiary amines, substituted amines, including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, etc. Certain organic amines include isopropylamine, benzathine, cholinate, diethanolamine, diethylamine, lysine, meglumine, piperazine and tromethamine.

In another aspect, the present invention provides compounds in the salt form of acetate, ascorbate, adipate, aspartate, benzoate, besylate, bromide/hydrobromide, bicarbonate/carbonate, bisulfate/sulfate, camphorsulfonate, capricate, chloride/hydrochloride, chlorotheophyllonate, citrate, ethanedisulfonate, fumarate, gluceptate, gluconate, glucuronate, glutamate, glutarate, glycolate, hippurate, hydroiodide/iodide, isethionate, lactate, lactobionate, lauryl sulfate, malate, maleate, malonate, mandelic acid salt, mesylate, methyl sulfate, mucinic acid salt, naphthoate, napsylate, nicotinate, nitrate, octadecanoate, oleate, oxalate, palmitate, pamoate, phosphate/hydrogenphosphate/dihydrogenphosphate, polygalacturonate, propionate, sebacate, stearate, succinate, sulfosalicylate, sulfate, tartrate, tosylate triphenate, trifluoroacetate or xinafoate.

Any formula given herein also implies the presence of unlabeled forms as well as isotopically labeled forms of the compounds. Isotopically labeled compounds have the structures depicted by the formulas given herein, except that one or more atoms are replaced by an atom having the selected atomic mass or mass number. Isotopes that can be included in the compounds of the present invention include, for example, isotopes of hydrogen.

In addition, the inclusion of certain isotopes, particularly deuterium (i.e., ^2H or D), may provide certain therapeutic benefits due to higher metabolic stability, such as increased in vivo half-life, or reduced dosage requirement, or improved therapeutic index or tolerability. It will be understood that deuterium is considered in this context as a substitute for the compound of the present invention. Deuterium concentration can be determined using the isotopic enrichment factor. As used herein, the term “isotopic enrichment factor” means the ratio of the abundance of an isotope to the natural abundance of that isotope. When a substituent in a compound of the present invention is specified as deuterium, such compound has an isotopic enrichment factor for each designated deuterium atom of at least 3500 (52.5% deuterium inclusion in each designated deuterium atom), at least 4000 (60% inclusion deuterium), at least 4500 (67.5% deuterium inclusion), at least 5000 (75% deuterium inclusion), at least 5500 (82.5% deuterium inclusion), at least 6000 (90% deuterium inclusion) , at least 6333.3 (95% deuterium inclusion), at least 6466.7 (97% deuterium inclusion), at least 6600 (99% deuterium inclusion) or at least 6633.3 (99.5% inclusion deuterium). It should be understood that the term "isotopic enrichment factor" can be used for any isotope in the same manner as described for deuterium.

Other examples of isotopes that can be included in the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, such as ³ H, ¹¹ C, ¹³ C, ¹⁴ C, ¹⁵ N, ¹⁸ F ³¹ P , ³² P, ³⁵ S, ³⁶ Cl, ¹²³ I, ¹²⁴ I, ¹²⁵ I, respectively. Therefore, it should be understood that the present invention includes compounds in which one or more of any of the above isotopes are included, including, for example, radioactive isotopes such as ³ H and ¹⁴ C, or compounds in which non-radioactive isotopes are present, such as ² H and ¹³ C. Such isotopically labeled compounds are useful in metabolism studies (using ¹⁴ C), reaction kinetics studies (for example, using ² H or ³ H), detection or imaging techniques such as positron emission tomography (PET) or single photon emission computed tomography (SPECT), including tests of drug or substrate distribution in tissues, or during radiation therapy of patients. In particular, ^{the 18} F or labeled compound may be particularly useful for PET or SPECT studies. The isotope-labeled compounds of the present invention can generally be prepared by conventional techniques known to those skilled in the art, or by methods similar to those described in the accompanying Examples and Preparation Methods, using a suitable isotope-labeled reagent instead of the unlabeled reagent used previously.

As used herein, the term “pharmaceutical composition” refers to a compound of the present invention or a pharmaceutically acceptable salt thereof together with at least one pharmaceutically acceptable carrier in a form suitable for oral or parenteral administration.

As used herein, the term "pharmaceutically acceptable carrier" refers to a substance useful in the preparation or use of a pharmaceutical composition and includes, for example, suitable diluents, solvents, dispersion media, surfactants, antioxidants, preservatives, isotonic agents, buffering agents, emulsifiers, absorption retarding agents, salts, drug stabilizers, binders, excipients, disintegrants, lubricants, wetting agents, sweeteners, flavoring agents, coloring agents and combinations thereof, which should be known to those skilled in the art (see eg Remington The Science and Practice of Pharmacy, ^22nd Ed. Pharmaceutical Press, 2013, pp. 1049-1070).

The term "therapeutically effective amount" of a compound of the present invention refers to an amount of a compound of the present invention that will produce a biological or medical response in a subject, such as reducing or inhibiting the activity of an enzyme or protein, or reducing the severity of symptoms, alleviating a condition, slowing or delaying the progression disease, or prevent disease, etc. In one embodiment, the term "therapeutically effective amount" refers to an amount of a compound of the present invention that, when administered to a subject, is effective in (1) at least partially alleviating, preventing, and/or reducing the severity of a condition or disorder or disease (i) mediated by the mTOR pathway, or (ii) associated with mTOR activity, or (iii) characterized by activity (normal or abnormal) of mTOR; or (2) reducing or suppressing mTOR activity; or (3) reduction or suppression of mTOR expression. In one embodiment, the term "therapeutically effective amount" refers to an amount of a compound of the present invention that, when administered to a cell or tissue or non-cellular biological material or environment, is effective to at least partially reduce or inhibit mTOR activity or at least to the extent of partial reduction or suppression of mTOR expression.

As used herein, the term "subject" means a primate (e.g. people, men or women), dogs, cats, rabbits, guinea pigs, pigs, rats and mice. In certain embodiments, the subject is a primate. In other embodiments, the subject is a human.

As used herein, the terms “administer,” “administration,” or “administration” refer to implantation, absorption, ingestion, injection, inhalation, or other administration of a compound of the present invention or a pharmaceutical composition based thereon.

As used herein, the terms “suppress,” “suppress,” or “suppress” mean a reduction or attenuation of a given condition, symptom, or disorder, or disease, or a significant reduction in the baseline activity of a biological activity or process.

As used herein, the term “treat,” “treat,” or “cure” any disease or disorder refers to alleviating, delaying the onset of, reducing the severity of the disease or disorder (i.e., slowing or stopping the progression of the disease or at least one of its clinical symptoms) or reduction or decrease in at least one physical parameter or biomarker associated with a disease or disorder, including those that may not be felt by the patient. In some embodiments, “treating,” “treating,” or “providing treatment” requires that signs or symptoms of a disease, disorder, or condition develop or be observed. In other embodiments, the treatment may be administered in the absence of signs or symptoms of the disease or condition. For example, the treatment may be administered to a susceptible individual prior to the onset of symptoms (eg, taking into account symptom history and/or taking into account genetic factors or other susceptibility factors). Treatment may also be continued after symptoms have resolved, for example to delay the onset or prevent relapse.

As used herein, the term “prevent,” “prevent,” or “prevent” any disease or disorder refers to prophylactically treating a disease or disorder or delaying the onset or progression of a disease or disorder.

As used herein, the term “age-related disease or disorder” refers to any disease or disorder whose incidence in a population or severity in an individual correlates with increasing age. More specifically, an age-related disease or disorder is a disease or disorder whose incidence is at least 1.5 times higher in people over 65 years of age compared to people aged 25-35 years. Examples of age-related disorders include, but are not limited to, sarcopenia, skin atrophy, types of senile hemangioma, types of seborrheic keratosis, brain atrophy, also called dementia, atherosclerosis, arteriosclerosis, emphysema, osteoporosis, osteoarthritis, high blood pressure, erectile dysfunction, types of cataracts, macular degeneration , glaucoma, stroke, cerebrovascular disease(s), chronic kidney disease, diabetes-associated kidney disease, liver dysfunction, liver fibrosis, autoimmune hepatitis, endometrial hyperplasia, metabolic disorder, renovascular disease, hearing loss, mobility impairment (eg , frailty), decreased cognitive ability, tendon stiffness, cardiac dysfunction such as cardiac hypertrophy and/or systolic and/or diastolic dysfunction and/or hypertension, cardiac dysfunction that leads to decreased ejection fraction, aging of the immune system, Parkinson's disease, Alzheimer's disease, cancer, aging of the immune system 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 including complications associated with diabetes, such as kidney failure, blindness and neuropathy.

As used herein, a subject is “in need of” treatment if, as a result of such treatment, such subject will benefit from a biological, medical point of view. or his quality of life will improve.

All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. Presented in this document is the use of all possible examples or introductory words before an example object (for example, "such as") is intended solely to better highlight the present invention and is not intended to limit the scope of the present invention as claimed elsewhere.

Any asymmetric atom (for example, carbon or the like) in the compound(s) of the present invention may be in racemic or enantiomerically enriched form, for example in (R)-, (S)- or (R, S)-configurations. In certain embodiments, each asymmetric atom is characterized by at least 50% enantiomeric excess, at least 60% enantiomeric excess, at least 70% enantiomeric excess, at least 80% enantiomeric excess, at least 90% enantiomeric excess, at least at least 95% enantiomeric excess or at least 99% enantiomeric excess in (R)- or (S)-configurations. Substituents on atoms with unsaturated double bonds may, if possible, be incis- (Z)- ortrance- (E)-form.

Accordingly, as used herein, a compound of the present invention may be in the form of one of possible stereoisomers, rotamers, atropisomers, tautomers, or mixtures thereof, such as substantially pure geometric ( cis or trans ) stereoisomers, diastereoisomers, optical isomers (antipodes), racemates or mixtures thereof.

Any resulting mixtures of stereoisomers can be separated based on the physicochemical differences of the constituents into pure or substantially pure geometric or optical isomers, diastereomers, racemates, for example, by chromatography and/or fractional crystallization.

Any resulting racemates of the compounds of the present invention or intermediates can be separated into optical antipodes by known methods, e.g. by separating their diastereomeric salts obtained with an optically active acid or base and isolating the optically active acid or base compounds. In particular, the basic fragment can thus be used to separate the compounds of the present invention into their optical antipodes, for example, by fractional crystallization of a salt formed with an optically active acid, e.g. tartaric acid, dibenzoyltartaric acid, diacetyltartaric acid, di-O, O'-P-toluoyltartaric acid, mandelic acid, malic acid or camphor-10-sulfonic acid. Racemic compounds of the present invention or racemic intermediates can also be separated by chiral chromatography, for example high pressure liquid chromatography (HPLC) using a chiral adsorbent.

Methods for preparing compounds of formula (I)

In another aspect, the present invention provides a method for preparing compounds of formula (I), formula (I)-A and formula (I)-B. The compounds of formula (I), formula (I)-A and formula (I)-B can be prepared in accordance with the following method described in schemes 1, 2 and 3 .

Scheme 1

A compound of formula (I) wherein R₂ defined for formula (I), can be prepared by reacting C32-deoxorapamycin (intermediate 1) with R₂-H, where R₂ defined for formula (I), in the presence of a suitable replacement reagent, e.g. p-toluenesulfonic acid, in the presence of a suitable solvent, e.g. dichloromethane. Suitable conditions are as follows:

1) R ₂ -H, p-toluenesulfonic acid-H ₂ O, dichloromethane, room temperature;

2) R ₂ -H, trifluoroacetic acid, -40°C, dichloromethane (see EP1212331B1);

3) R ₂ -H, 5 M LiClO ₄ , Et ₂ O (0.1 M), room temperature (see TL, 1995, 43, 7823);

4) R ₂ -H, Cp ₂ HfCl ₂ -AgClO ₄ (Suzuki catalyst), 4A MS, dichloromethane, room temperature (see TL, 1995, 43, 7823);

5) R ₂ -H, BF ₃ -OEt ₂ or Zn(OTf) ₂ , THF, 0°C (see TL, 1994, 37, 6835);

6) R ₂ -H, ZnCl ₂ , dichloromethane, 0°C (see JOC, 1994, 59, 6512).

C32-deoxorapamycin used as a starting material can be obtained by methods known in the art, for example, as described in US Patent Publication No. 005985890orWO 2007085400.

Scheme 2

A compound of formula (I)-A, where R₁ selected from and R₂ defined for formula (I), can be prepared by reacting intermediate 1 with R₁-H or R₁-X followed by reaction with R₂-H. In one embodiment, intermediate 1 is reacted with R₁-H or R₁-X under alkylation, phosphinating or esterification conditions to give intermediate 1-A. In one embodiment, intermediate 1-A is reacted with R₂-H under substitution reaction conditions, e.g. as presented herein, to obtain the compound of formula (I)-A.

Scheme 3

A compound of formula (I)-C, where R₁ represents; and R₂ defined for formula (I), can be prepared by reacting intermediate 1 with R₁-H followed by reaction with R₂-H. In one embodiment, intermediate 1 is activated and reacted under nucleophilic conditions to produce intermediate 1-A. In one embodiment, intermediate 1-A is reacted with R₂-H under substitution reaction conditions, for example, as presented herein, to obtain the compound of formula (I)-C.

The reactions can be carried out according to conventional methods, for example as described in the examples.

Workup of the reaction mixtures and purification of the compounds thus obtained can be carried out in accordance with known procedures.

Acid addition salts can be prepared from free bases in a known manner and vice versa.

The starting materials may be known or prepared according to conventional procedures from known compounds, for example as described in the Examples.

In another aspect, the present invention provides a pharmaceutical composition comprising a compound of the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. In a further embodiment, the composition contains at least two such pharmaceutically acceptable carriers as described herein. The pharmaceutical composition may be formulated for specific routes of administration, such as oral administration, parenteral administration (e.g. by injection, infusion, transdermal or topical administration) and rectal administration. Topical administration may also refer to inhalation or intranasal administration. The pharmaceutical compositions of the present invention can be formulated in solid form (including, but not limited to, capsules, tablets, pills, granules, powders, or suppositories) or liquid form (including, without limitation, solutions, suspensions, or emulsions). Tablets can be either film-coated or enteric-coated according to methods known in the art. Typically, pharmaceutical compositions are tablets or gelatin capsules containing the active ingredient together with one or more of:

a) thinners, for example lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and/or glycine;

b) lubricants, for example, silicon dioxide, talc, stearic acid, its magnesium or calcium salt and/or polyethylene glycol; also in case of tablets

c) binders, for example magnesium aluminosilicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidone; if necessary

d) leavening agents, for example, types of starch, agar, alginic acid or its sodium salt or effervescent mixtures; And

e) absorbents, dyes, flavors and sweeteners.

The compound of the present invention or a pharmaceutically acceptable salt thereof may also be in the form of a drug-eluting stent, ie, a stent coated with a compound of the present invention or a pharmaceutically acceptable salt thereof.

The compounds of the present invention, in free form or in pharmaceutically acceptable salt form, exhibit valuable pharmacological properties, e.g. modulating properties on the mTOR pathway, e.g. shown by analysisin vitro Andin vivo, presented in the following sections, and are therefore intended for therapy or for use as research chemicals, e.g. as pharmacologically active compounds.

Methods for measuring the activity of mTORC1 inhibitors are widely known in the art. Typically, activity is determined using IC50 values, which are assessed by determining the degree of inhibition of S6 phosphorylation, which is present in the mTORC1 signaling pathway. IC50 values for mTORC1 inhibitors are compared with the IC50 value for rapamycin in the same assay. An mTORC1 inhibitor having an IC50 value within 100 times the IC50 value of rapamycin in the same assay is suitable for use in the present invention, meaning that a less active rapalog may still be needed to more easily achieve only partial inhibition of mTORC1 activity under certain conditions. conditions, as well as to improve the ability to measure the level of a molecule in the bloodstream (since higher concentrations are required for a less active molecule), which would be useful for fine-tuning the concentration/potency ratio in the blood.

Suitable assays for measuring the activity of mTOR inhibitors are described, for example, in US Pat. No. 5,665,772, as measured by IC50 value in an MLR (mixed lymphocyte culture reaction) assay and/or in an IL-6 (interleukin-6)-dependent proliferation assay.

MLR analysis is typically carried out as follows. Spleen cells (0.5 × 106) from Balb/c mice (female, 8-10 weeks) were incubated for 5 days with irradiated (2000 rad), mitomycin C-treated spleen cells (0.5 × 106) from CBA mice (females, 8-10 weeks). Irradiated allogeneic cells induce a proliferative response in Balb/c spleen cells, which can be measured by incorporation of a labeled precursor into DNA. Because the stimulator cells are irradiated (or treated with mitomycin C), they do not respond to Balb/c cells with proliferation, but retain their antigenicity. The antiproliferative effect of test compounds on Balb/c cells is measured at various dilutions and the concentration resulting in 50% inhibition of cell proliferation (IC50) is calculated. The inhibitory ability of a test sample can be compared to rapamycin and expressed as a relative IC50 value (i.e., test sample IC50/rapamycin IC50).

The IL-6-mediated proliferation assay is typically performed as follows: The assay uses an interleukin-6 (IL-6)-dependent mouse hybridoma cell line and is performed in 96-well microtiter plates. 5000 cells/well are cultured in serum-free medium (as described by M. H. Schreier and R. Tees in Immunological Methods, I. Lefkovits and B. Pernis, eds., Academic Press 1981. Vol. II, pp. 263-275) supplemented with 1 ng recombinant IL-6/ml. After 66 hours of incubation in the absence or presence of the test sample, cells are pulsed with 1 μCi (3-H)-thymidine/well for an additional 6 hours, collected and counted by liquid scintillation. Incorporation of (3-H)-thymidine into DNA correlates with an increase in cell number and is thus a measure of cell proliferation. Serial dilution of the test sample allows the concentration resulting in 50% inhibition of cell proliferation (IC50) to be calculated. The inhibitory ability of a test sample can be compared to rapamycin and expressed as a relative IC50 value (i.e., test sample IC50/rapamycin IC50).

The activity of mTOR inhibitors can also be determined using a cell-based assay using TSC1-/- MEF cells. TSC1−/− MEF cells are mouse embryonic fibroblasts deficient in the tuberous sclerosis protein TSC1, which negatively regulates mTORC1 signaling. Thus, TSC1 deficiency induces constitutive activation of mTORC1, which leads to phosphorylation (activation) of proteins downstream in the mTORC1 signaling pathway. This cell-based assay is used to measure inhibition (dephosphorylation) of components S6 and 4EBP1 of the mTORC1 signaling pathway by rapalogues or other mTOR inhibitors.

The analysis is usually carried out as follows. TSC1−/− MEF cells were seeded into poly-D-lysine-coated clear-bottom Greiner 384-well plates and incubated overnight at 37°C, 5% CO2. The next day, the cells are washed 8 times with a “severe starvation” solution (1 L DPBS + 1 g D (+)-glucose + 10 ml 7.5% sodium bicarbonate solution + 20 ml 1 M HEPES) and incubated for another 2 hours in the same solution. Cells are then treated with compounds at decreasing concentrations (8 levels at 3.16-fold dilution) and incubated for 2 hours at 37°C, 5% CO2. Cells are fixed with 4% paraformaldehyde for 30 min. and washed 5 times with TBS-EDTA followed by immunostaining with fluorescently labeled antibodies to pS6 and p4EBP1. The nuclei are made visible using Hoechst dye. Cells are visualized using the appropriate fluorescent channels and the activity of mTOR inhibitors is determined using IC ₅₀ (nM) against pS6.

Diseases and disorders

The compounds of the present invention may be useful in the prevention or treatment of a symptom or prodrome selected from:

- acute or chronic organ or tissue transplant rejection;

- graft-related vasculopathies;

- proliferation and migration of smooth muscle cells, leading to thickening of the vascular intima, blockage of blood vessels, obstructive coronary atherosclerosis, restenosis;

- autoimmune diseases and inflammatory conditions;

- diseases associated with the treatment and prevention of asthma;

- diseases associated with multiple drug resistance (MDR);

- infections caused by fungi;

- inflammation;

- infections;

- age-related diseases;

- neurodegenerative diseases;

- proliferative disorders, in particular cancer;

- epileptic seizures and disorders associated with epileptic seizures;

- mitochondrial myopathy and mitochondrial stress;

- treatable conditions that have been shown to increase the likelihood of age-related diseases, such as conditions in which there is an increase in the levels of cytokines that cause aging (for example, IL6);

- disorders that involve a process of fibrosis and/or inflammation, e.g. liver and kidney disorders. Examples include liver fibrosis, which occurs in end-stage liver disease; cirrhosis of the liver; liver failure caused by toxicity; non-alcoholic steatohepatitis or NASH and alcoholic steatosis. Another example is renal fibrosis, which occurs as a result of acute kidney injury leading to chronic kidney disease. Also, diabetic nephropathy can induce renal fibrosis and inflammation. Often, kidney disease causes heart failure as a result of increased blood pressure; it may also be associated with cardiac fibrosis. Rapalogs have preclinical efficacy in models of heart failure and are effective in reducing liver fibrosis in liver transplant patients (Buss, S.J.et al. Beneficial effects of Mammalian target of rapamycin inhibition on left ventricular remodeling after myocardial infarction. J Am Coll Cardiol. (2009) 54(25): 2435-46; Buss, S.J.et al. Augmentation of autophagy by mTOR-inhibition in myocardial infarction: When size matters. Autophagy. (2010) 6(2):304-6; Villamil, F. G.et al. Fibrosis progression in maintenance liver transplant patients with hepatitis C recurrence: a randomized study of everolimus vs. calcineurin inhibitors. Liver Int. (2014) 34(10):1513-21).

Treatment of acute or chronic organ or tissue transplant rejection includes treatment of recipients, e.g. heart, lung, heart and lung, liver, kidney, pancreas, skin or cornea transplants. The compounds of the present invention are also intended to prevent graft-versus-host disease, for example, after bone marrow transplantation.

Graft-associated vasculopathies include atherosclerosis.

Autoimmune diseases and inflammatory conditions include, but are not limited to, inflammatory conditions with an etiology that includes an autoimmune component, such as arthritis (eg, rheumatoid arthritis, progressive chronic arthritis, and arthritis deformans) and rheumatic diseases. Specific autoimmune diseases for which compounds of formula (I), formula (I)-A, formula (I)-B and formula (I)-C can be used include autoimmune hematological disorders (including, for example, hemolytic anemia, aplastic anemia, congenital aplastic anemia and idiopathic thrombocytopenia), systemic lupus erythematosus, polychondria, scleroderma, Wegener's granulomatosis, dermatomyositis, chronic active hepatitis, myasthenia gravis, psoriasis, Stevens-Johnson syndrome, idiopathic malabsorption syndrome, autoimmune inflammatory bowel disease (including, for example, ulcerative colitis and Crohn's disease), endocrine ophthalmopathy, Graves' disease, sarcoidosis, multiple sclerosis, primary biliary cirrhosis, juvenile diabetes (type I diabetes), uveitis (anterior and posterior), dry eye syndrome and vernal keratoconjunctivitis, interstitial pulmonary fibrosis, psoriatic arthritis, glomerulonephritis (with and without nephrotic syndrome, e.g. including idiopathic nephrotic syndrome or minimal change nephropathy) and juvenile dermatomyositis.

Treatment for multidrug resistance (MDR) involves improving the effectiveness of other chemotherapeutic agents in treating and controlling multidrug-resistant conditions such as multidrug-resistant cancer or multidrug-resistant AIDS. MDR is especially problematic in cancer patients and ADS patients who do not respond to traditional chemotherapy because Pgp clears the drug from cells.

Infection includes infection caused by pathogens having Mip factors or Mip-like factors.

Age-related diseases include sarcopenia, skin atrophy, types of senile hemangioma, types of seborrheic keratosis, brain atrophy, also called dementia, atherosclerosis, arteriosclerosis, emphysema, osteoporosis, osteoarthritis, high blood pressure, erectile dysfunction, types of cataracts, macular degeneration, glaucoma, stroke, cerebrovascular disease(s), chronic kidney disease, diabetes-associated kidney disease, liver dysfunction, liver fibrosis, autoimmune hepatitis, endometrial hyperplasia, metabolic disorder, renovascular disease, hearing loss, mobility impairment (eg, frailty), decreased cognitive ability, tendon stiffness, cardiac dysfunction such as cardiac hypertrophy and/or systolic and/or diastolic dysfunction and/or hypertension, cardiac dysfunction that leads to decreased ejection fraction, aging of the immune system, Parkinson's disease, Alzheimer's disease, cancer, aging of the immune system 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 including complications associated with diabetes, such as kidney failure, blindness and neuropathy.

Neurodegenerative diseases include Huntington's disease, Parkinson's disease, spinocerebellar ataxia type 3, Alzheimer's disease, motor neuron disease, and peripheral neuropathy.

Proliferative disorders include cancer. Such conditions include those listed in US Pat. No. 9,669,032, such as kidney cancer, renal cell carcinoma, colorectal cancer, uterine sarcoma, uterine endometrial cancer, endometrial cancer, breast cancer, ovarian cancer, cervical cancer, gastric cancer, fibrosarcoma , pancreatic cancer, liver cancer, melanoma, leukemia, multiple myeloma, nasopharyngeal cancer, prostate cancer, lung cancer, glioblastoma, bladder cancer, mesothelioma, head cancer, rhabdomyosarcoma, sarcoma, lymphoma or neck cancer.

Epileptic seizures and seizure-related disorders include West syndrome, focal cortical dysplasia (FCD), tuberous sclerosis complex (TSC), childhood absence epilepsy, benign focal childhood epilepsy, juvenile myoclonic epilepsy (JME), temporal lobe epilepsy, frontal lobe epilepsy, refractory epilepsy, Lennox-Gastaut syndrome, occipital lobe epilepsy, Proteus syndrome, hemimegalencephaly syndrome (HMEG), megalencephaly syndrome (MEG), megalencephaly -capillary malformation (MCAP) and megalencephaly -polymicrogyria-polydactyly-hydrocephalus syndrome ( MPPH).

Mitochondrial myopathy and mitochondrial stress are mitochondrial disorders described in Chinnery, P. F. (2015); EMBO Mol. Med. 7, 1503-1512; Koopman, W. J. et al. , (2016); EMBO Mol. Med. 8, 311-327 and Young, M. J., and Yound and Copeland, W. C. (2016); Curr. Opin. Genet. Dev. 38, 52-62.

Treatable conditions that have been shown to increase the likelihood of age-related diseases include aging, e.g. aging of the immune system. It is diagnosed by (i) increased levels of circulating cytokines such as IL-6, but also by (ii) senescent cells found in muscle, kidney, liver, brain, neurons, liver, pancreas or heart; or also by (iii) a decrease in the efficiency of DNA repair, which can be demonstrated by an increase in the level of transcription of repetitive elements, including genes encoded by a transposon. For background, see Baker, D. J. et al, Nature, 2016; 530(7589):184-9. doi: 10.1038/nature16932. Epub 2016 Feb 3.

Treatment methods and applications

The present invention provides the use of a compound of the present invention for use in therapy. In a further embodiment, the therapy is selected for a disease or disorder or concomitant disease that can be treated by modulation mTOR pathway.In one embodiment, the disease is selected from the above list, in one embodiment, an age-related disease, such as morbidity associated with respiratory tract infection in the elderly.

The present invention provides a compound of the present invention for use in therapy. In a further embodiment, the therapy is selected for a disease that can be treated by modulation mTOR pathways. In one embodiment, the disease is selected from the above list, in one embodiment, an age-related disease, such as morbidity associated with respiratory tract infection in the elderly.

The present invention provides for the use of a compound of the present invention for the manufacture of a medicinal product. In a further embodiment, the medicinal product is for the prevention or treatment of a disease that can be treated by modulation mTOR pathways. In one embodiment, the disease is selected from the above list, in one embodiment, an age-related disease, such as morbidity associated with respiratory tract infection in the elderly.

In one aspect, the present invention provides a method of treating a disorder or disease mediated by the mTOR pathway in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of Formula (I), Formula (I)-A, Formula (I) -B and formula (I)-C or a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing a compound of formula (I), formula (I)-A, formula (I)-B and formula (I)-C or a pharmaceutically acceptable salt thereof , or a pharmaceutical combination containing a compound of formula (I), formula (I)-A, formula (I)-B and formula (I)-C or a pharmaceutically acceptable salt thereof.

In one aspect, the present invention provides a method of treating a disease or disorder in a subject, wherein the target tissue or organ associated with pathology associated with the disease or disorder has levels of FKBP12 sufficient to inhibit mTORC1, the method comprising administering to the subject in need herein, a therapeutically effective amount of a compound of formula (I), formula (I)-A, formula (I)-B and formula (I)-C, or a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing a compound of formula (I), formula ( I)-A, formula (I)-B and formula (I)-C or a pharmaceutically acceptable salt thereof, or a pharmaceutical combination containing a compound of formula (I), formula (I)-A, formula (I)-B and formula (I)-C or a pharmaceutically acceptable salt thereof.

In one aspect, the present invention provides a method of treating a disease or disorder in a subject having, or previously determined to have, levels of FKBP12 sufficient to inhibit mTORC1, the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of formula (I), formula (I)-A, formula (I)-B and formula (I)-C or a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing a compound of formula (I), formula (I)-A, formula (I)-B and formula (I)-C or a pharmaceutically acceptable salt thereof, or a pharmaceutical combination containing a compound of formula (I), formula (I)-A, formula (I)-B and formula (I)-C or a pharmaceutically acceptable salt thereof.

In one embodiment, the disease or disorder is selected from sarcopenia, skin atrophy, types of senile hemangioma, types of seborrheic keratoses, cerebral atrophy, atherosclerosis, arteriosclerosis, emphysema, osteoporosis, osteoarthritis, high blood pressure, erectile dysfunction, types of cataracts, macular degeneration, glaucoma, stroke, cerebrovascular disease(s), chronic kidney disease, diabetes-associated kidney disease, liver dysfunction, liver fibrosis, autoimmune hepatitis, endometrial hyperplasia, metabolic disorder, renovascular disease, hearing loss, mobility impairment, cognitive decline abilities, tendon stiffness, cardiac dysfunction such as cardiac hypertrophy and/or systolic and/or diastolic dysfunction and/or hypertension, cardiac dysfunction that results in decreased ejection fraction, aging of the immune system, Parkinson's disease, Alzheimer's disease, cancer, aging of the immune system 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 associated with diabetes, such as kidney failure, blindness and neuropathy.

In one embodiment, the disorder is liver fibrosis.

In one aspect, the present invention provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of Formula (I), Formula (I)-A, Formula (I)-B, and Formula ( I)-C or a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing a compound of formula (I), formula (I)-A, formula (I)-B and formula (I)-C or a pharmaceutically acceptable salt thereof, or a pharmaceutical combination containing a compound of formula (I), formula (I)-A, formula (I)-B and formula (I)-C, or a pharmaceutically acceptable salt thereof, wherein the disorder or disease is selected from

- acute or chronic organ or tissue transplant rejection;

- graft-related vasculopathies;

- proliferation and migration of smooth muscle cells, leading to thickening of the vascular intima, blockage of blood vessels, obstructive coronary atherosclerosis, restenosis;

- autoimmune diseases and inflammatory conditions;

- diseases associated with the treatment and prevention of asthma;

- diseases associated with multiple drug resistance (MDR);

- infections caused by fungi;

- inflammation;

- infections;

- age-related diseases;

- neurodegenerative diseases;

- proliferative disorders, in particular cancer;

- epileptic seizures and disorders associated with epileptic seizures and

- mitochondrial myopathy and mitochondrial stress.

In one embodiment, the disorder is a disorder that involves the process of fibrosis and/or inflammation.

In one embodiment, the disorder is selected from liver and kidney disorders.

In one embodiment, the liver disorder is selected from liver fibrosis, which occurs in end-stage liver disease; liver cirrhosis; liver failure caused by toxicity; non-alcoholic steatohepatitis or NASH and alcoholic steatosis.

In one embodiment, the kidney disorder is renal fibrosis.

In one embodiment, renal fibrosis occurs as a result of acute kidney injury.

In one embodiment, the kidney disorder is a chronic kidney disorder.

In one embodiment, the kidney disorder is diabetic nephropathy.

In one aspect, the present invention provides a method of treating an age-related disorder or disease in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of Formula (I), Formula (I)-A, Formula (I)-B, and Formula (I)-C or a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing a compound of formula (I), formula (I)-A, formula (I)-B and formula (I)-C or a pharmaceutically acceptable salt thereof, or a pharmaceutical combination , containing a compound of formula (I), formula (I)-A, formula (I)-B and formula (I)-C or a pharmaceutically acceptable salt thereof, wherein the disorder or disease is selected from sarcopenia, skin atrophy, types of senile hemangioma, types seborrheic keratosis, cerebral atrophy, atherosclerosis, arteriosclerosis, pulmonary emphysema, osteoporosis, osteoarthritis, high blood pressure, erectile dysfunction, types of cataracts, macular degeneration, glaucoma, stroke, cerebrovascular disease(s), chronic kidney disease, diabetes-associated kidney disease , liver dysfunction, liver fibrosis, autoimmune hepatitis, endometrial hyperplasia, metabolic disorders, renovascular disease, hearing loss, impaired ambulation, cognitive decline, tendon stiffness, cardiac dysfunction such as cardiac hypertrophy, and/or systolic and/ or diastolic dysfunction and/or hypertension, cardiac dysfunction that leads to decreased ejection fraction, aging of the immune system, Parkinson's disease, Alzheimer's disease, cancer, aging of the immune system 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 diabetes-related complications such as kidney failure, blindness and neuropathy.

In one aspect, the present invention provides a method of treating cancer in a subject, the method comprising administering to the subject a therapeutically effective amount of a compound of Formula (I), Formula (I)-A, Formula (I)-B, and Formula (I)-C, or a pharmaceutically acceptable salt, a pharmaceutical composition containing a compound of formula (I), formula (I)-A, formula (I)-B and formula (I)-C or a pharmaceutically acceptable salt thereof, or a pharmaceutical combination containing a compound of formula (I) , formula (I)-A, formula (I)-B and formula (I)-C, or a pharmaceutically acceptable salt thereof.

In one embodiment, the method further comprises the use of a PD-1/PDL-1 inhibitor.

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

In one aspect, the present invention provides a compound of Formula (I), Formula (I)-A, Formula (I)-B, and Formula (I)-C, or a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing a compound of Formula (I), Formula (I)-A, formula (I)-B and formula (I)-C or a pharmaceutically acceptable salt thereof, or a pharmaceutical combination containing a compound of formula (I), formula (I)-A, formula (I)-B and formula (I)-C or a pharmaceutically acceptable salt thereof, for use as a medicinal product.

In one aspect, the present invention provides a compound of Formula (I), Formula (I)-A, Formula (I)-B, and Formula (I)-C, or a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing a compound of Formula (I), Formula (I)-A, formula (I)-B and formula (I)-C or a pharmaceutically acceptable salt thereof, or a pharmaceutical combination containing a compound of formula (I), formula (I)-A, formula (I)-B and formula (I)-C or a pharmaceutically acceptable salt thereof, for use in the prevention or treatment of a disorder or disease mediated by the mTOR pathway.

In one aspect, the present invention provides a compound of Formula (I), Formula (I)-A, Formula (I)-B, and Formula (I)-C, or a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing a compound of Formula (I), Formula (I)-A, formula (I)-B and formula (I)-C or a pharmaceutically acceptable salt thereof, or a pharmaceutical combination containing a compound of formula (I), formula (I)-A, formula (I)-B and formula (I)-C or a pharmaceutically acceptable salt thereof, for use in the prevention or treatment of a disorder or disease selected from

- acute or chronic organ or tissue transplant rejection;

- graft-related vasculopathies;

- proliferation and migration of smooth muscle cells, leading to thickening of the vascular intima, blockage of blood vessels, obstructive coronary atherosclerosis, restenosis;

- autoimmune diseases and inflammatory conditions;

- diseases associated with the treatment and prevention of asthma;

- diseases associated with multiple drug resistance (MDR);

- infections caused by fungi;

- inflammation;

- infections;

- age-related diseases;

- neurodegenerative diseases;

- proliferative disorders, in particular cancer;

- epileptic seizures and disorders associated with epileptic seizures;

- mitochondrial myopathy and mitochondrial stress and

- treatable conditions that have been shown to increase the likelihood of age-related diseases, such as conditions in which there is an increase in the levels of cytokines that cause aging.

In one aspect, the present invention provides a compound of Formula (I), Formula (I)-A, Formula (I)-B, and Formula (I)-C, or a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing a compound of Formula (I), Formula (I)-A, formula (I)-B and formula (I)-C or a pharmaceutically acceptable salt thereof, or a pharmaceutical combination containing a compound of formula (I), formula (I)-A, formula (I)-B and formula (I)-C or a pharmaceutically acceptable salt thereof, for use in the prevention or treatment of a disorder or disease that involves a process of fibrosis and/or inflammation.

In one embodiment, the disorder is selected from liver and kidney disorders.

In one embodiment, the liver disorder is selected from liver fibrosis, which occurs in end-stage liver disease; liver cirrhosis; liver failure caused by toxicity; non-alcoholic steatohepatitis or NASH and alcoholic steatosis.

In one embodiment, the kidney disorder is renal fibrosis, which occurs as a result of acute kidney injury.

In one embodiment, the kidney disorder is a chronic kidney disorder.

In one embodiment, the kidney disorder is diabetic nephropathy.

In one aspect, the present invention provides a compound of Formula (I), Formula (I)-A, Formula (I)-B, and Formula (I)-C, or a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing a compound of Formula (I), Formula (I)-A, formula (I)-B and formula (I)-C or a pharmaceutically acceptable salt thereof, or a pharmaceutical combination containing a compound of formula (I), formula (I)-A, formula (I)-B and formula (I)-C or a pharmaceutically acceptable salt thereof, for use in the prevention or treatment of an age-related disorder or disease selected from sarcopenia, skin atrophy, types of senile hemangioma, types of seborrheic keratosis, cerebral atrophy, also called dementia, atherosclerosis, arteriosclerosis, emphysema, osteoporosis, osteoarthritis, high blood pressure, erectile dysfunction, types of cataracts, macular degeneration, glaucoma, stroke, cerebrovascular disease(s), chronic kidney disease, diabetes-associated kidney disease, liver dysfunction, liver fibrosis, autoimmune hepatitis, endometrial hyperplasia, metabolic disorders, renovascular disease, hearing loss, impaired mobility (eg, frailty), decreased cognitive ability, tendon stiffness, cardiac dysfunction such as cardiac hypertrophy and/or systolic and/or diastolic dysfunction and/or hypertension, cardiac dysfunction that leads to decreased ejection fraction, aging of the immune system, Parkinson's disease, Alzheimer's disease, cancer, aging of the immune system 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 associated with diabetes, such as kidney failure, blindness and neuropathy.

In one aspect, the present invention provides a compound of Formula (I), Formula (I)-A, Formula (I)-B, and Formula (I)-C, or a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing a compound of Formula (I), Formula (I)-A, formula (I)-B and formula (I)-C or a pharmaceutically acceptable salt thereof, or a pharmaceutical combination containing a compound of formula (I), formula (I)-A, formula (I)-B and formula (I)-C or a pharmaceutically acceptable salt thereof, for use in the prevention or treatment of cancer.

In one aspect, the present invention provides a compound of Formula (I), Formula (I)-A, Formula (I)-B, and Formula (I)-C, or a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing a compound of Formula (I), Formula (I)-A, formula (I)-B and formula (I)-C or a pharmaceutically acceptable salt thereof, or a pharmaceutical combination containing a compound of formula (I), formula (I)-A, formula (I)-B and formula (I)-C or a pharmaceutically acceptable salt thereof, for use in the treatment of kidney cancer, renal cell carcinoma, colorectal cancer, uterine sarcoma, uterine endometrial cancer, endometrial cancer, breast cancer, ovarian cancer, cervical cancer, stomach cancer , fibrosarcoma, pancreatic cancer, liver cancer, melanoma, leukemia, multiple myeloma, nasopharyngeal cancer, prostate cancer, lung cancer, glioblastoma, bladder cancer, mesothelioma, head cancer, rhabdomyosarcoma, sarcoma, lymphoma or neck cancer.

In one embodiment, the present invention provides a connection

or a pharmaceutically acceptable salt thereof, for use in the treatment of an age-related disease, such as morbidity associated with respiratory tract infection in the elderly. In one embodiment, the compound is (S)-C16-(1,1-dioxidoisothiazolidin-2-yl)-C32-deoxorapamycin (Example 1) or a pharmaceutically acceptable salt thereof. In one embodiment, the compound is (R)-C16-(1,1-dioxidoisothiazolidin-2-yl)-C32-deoxorapamycin (Example 2) or a pharmaceutically acceptable salt thereof.

In one embodiment, the disorder or disease is selected from

- acute or chronic organ or tissue transplant rejection;

- graft-related vasculopathies;

- proliferation and migration of smooth muscle cells, leading to thickening of the vascular intima, blockage of blood vessels, obstructive coronary atherosclerosis, restenosis;

- autoimmune diseases and inflammatory conditions;

- diseases associated with the treatment and prevention of asthma;

- diseases associated with multiple drug resistance (MDR);

- infections caused by fungi;

- inflammation;

- infections;

- age-related diseases;

- neurodegenerative diseases;

- proliferative disorders, in particular cancer;

- epileptic seizures and disorders associated with epileptic seizures;

- mitochondrial myopathy and mitochondrial stress and

- treatable conditions that have been shown to increase the likelihood of age-related diseases, such as conditions in which there is an increase in the levels of cytokines that cause aging (for example, IL6).

In one embodiment, the disorder or disease is an age-related disorder or disease selected from sarcopenia, skin atrophy, senile hemangioma types, seborrheic keratosis types, cerebral atrophy, also called dementia, atherosclerosis, arteriosclerosis, emphysema, osteoporosis, osteoarthritis, high blood blood pressure, erectile dysfunction, types of cataracts, macular degeneration, glaucoma, stroke, cerebrovascular disease(s), chronic kidney disease, diabetes-associated kidney disease, liver dysfunction, liver fibrosis, autoimmune hepatitis, endometrial hyperplasia, metabolic disorders, renovascular disease , hearing loss, mobility impairment (for example, frailty), decreased cognitive ability, tendon stiffness, cardiac dysfunction such as cardiac hypertrophy and/or systolic and/or diastolic dysfunction and/or hypertension, cardiac dysfunction that leads to decreased ejection fraction, aging of the immune system, Parkinson's disease, Alzheimer's disease, cancer, aging of the immune system 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 associated with diabetes, such as kidney failure, blindness and neuropathy.

In one embodiment, the disorder or disease involves a process of fibrosis and/or inflammation, e.g. liver and kidney disorders. In one embodiment, the disorder is a kidney disorder. In one embodiment, the disorder is a liver disorder. Examples include liver fibrosis, which occurs during end-stage liver disease; cirrhosis of the liver; liver failure caused by toxicity; non-alcoholic steatohepatitis or NASH and alcoholic steatosis. Another example is renal fibrosis, which occurs as a result of acute kidney injury leading to chronic kidney disease. Also, diabetic nephropathy can induce renal fibrosis and inflammation. Often, kidney disease causes heart failure as a result of increased blood pressure; it may also be associated with cardiac fibrosis. Rapalogues have preclinical efficacy in heart failure models and are effective in reducing liver fibrosis in liver transplant patients.

The therapeutically effective dose of the compound, pharmaceutical composition, or combinations thereof depends on the type of subject, body weight, age, and individual condition, disorder, or disease being treated or the severity thereof. A qualified physician, clinician, or veterinarian can readily determine the effective amount of each active ingredient needed to prevent, treat, or inhibit the progression of the disorder or disease.

The above dosage parameters are evident in the testsin vitroAndin vivousing predominantly mammals, e.g. mice, rats, dogs, non-human apes or isolated organs, tissues and their preparations. The compounds of the present invention can be usedin vitro in the form of solutions, for example aqueous solutions, andin vivo either enterally or parenterally, preferably intravenously, e.g. in the form of a suspension or aqueous solution. Dosein vitromay range in molar concentration from approximately 10^-3 mol/l up to 10^-9 mol/l. Therapeutically effective amountin vivodepending on the route of administration, may be in the range of approximately 0.1-500 mg/kg or approximately 0.1-500 mg per subject.

The compound of the present invention can be administered either simultaneously with one or more other therapeutic agents, either before or after their administration. The compound of the present invention can be administered separately through the same or different route of administration or together in the same pharmaceutical composition as other agents. A therapeutic agent is, for example, a chemical compound, peptide, antibody, antibody fragment or nucleic acid that is therapeutically active or enhances therapeutic activity when administered to a patient in combination with a compound of the present invention.

In one embodiment, the present invention provides a product containing a compound of the present invention and at least one other therapeutic agent in the form of a combination preparation for simultaneous, separate or sequential use in therapy. In one embodiment, the therapy is the treatment of a disease or condition through partial or complete inhibition of mTOR. Products presented as a combination preparation include a composition containing a compound of the present invention and other therapeutic agent(s) together in the same pharmaceutical composition, or a compound of the present invention and other therapeutic agent(s). remedy(s) in a separate form, e.g. in the form of a set.

In one embodiment, the present invention provides a pharmaceutical composition comprising a compound of the present invention and other therapeutic agent(s). Optionally, the pharmaceutical composition may contain a pharmaceutically acceptable carrier as described above.

In one embodiment, the present invention provides a kit containing two or more separate pharmaceutical compositions, at least one of which contains a compound of the present invention. In one embodiment, the kit contains means for containing the compositions separately, such as a container, a divided bottle, or a divided foil pouch. An example of such a set is blister packaging, usually used for packaging tablets, capsules, etc.

The kit of the present invention can be used to administer different dosage forms, for example, for oral and parenteral use, to administer individual compositions at different dosage intervals, or to titrate individual compositions against each other. To promote compliance with a treatment regimen, the kit of the present invention typically contains instructions for administration.

In the combination therapies of the present invention, the compound of the present invention and the other therapeutic agent may be manufactured and/or formulated by the same or different manufacturers. Moreover, a compound of the present invention and another therapeutic agent can be combined in a combination therapy: (i) before the combination product reaches physicians (e.g. in the case of a kit containing a compound of the present invention and another therapeutic agent); (ii) by the physicians themselves (or under the supervision of a physician) shortly before administration; (iii) in the patients themselves, for example, during sequential administration of a compound of the present invention and another therapeutic agent.

Accordingly, the present invention provides the use of a compound of the present invention for the prevention or treatment of a disease or condition through partial or complete inhibition of mTOR, wherein the drug is formulated for administration with another therapeutic agent. The present invention also provides for the use of another therapeutic agent for the prevention or treatment of a disease or condition mediated by mTOR inhibition, wherein the drug is administered with a compound of the present invention.

The present invention also provides a compound of formula (I), formula (I)-A and formula (I)-B, or a pharmaceutically acceptable salt thereof, for use in a method of preventing or treating a disease or condition mediated by mTOR inhibition, wherein the compound of the present invention is produced for administration with another therapeutic agent. The present invention also provides another therapeutic agent for use in a method of preventing or treating a disease or condition mediated by mTOR inhibition, wherein the other therapeutic agent is prepared for administration with a compound of formula (I), formula (I)-A and formula (I)-B or a pharmaceutically acceptable salt thereof. The present invention also provides a compound of formula (I), formula (I)-A and formula (I)-B, or a pharmaceutically acceptable salt thereof, for use in a method of preventing or treating a disease or condition mediated by mTOR inhibition, wherein the compound is administered with another therapeutic means. The present invention also provides another therapeutic agent for use in a method of preventing or treating a disease or condition mediated by mTOR inhibition, wherein the other therapeutic agent is administered with a compound of formula (I), formula (I)-A and formula (I)-B, or pharmaceutically acceptable salt.

The present invention also provides the use of a compound of formula (I), formula (I)-A and formula (I)-B, or a pharmaceutically acceptable salt thereof, for the prevention or treatment of an mTOR-mediated disease or condition where the patient has previously (e.g. within 24 hours) was treated with another therapeutic agent. The present invention also provides for the use of another therapeutic agent for the prevention or treatment of an mTOR-mediated disease or condition where the patient has previously (eg, for 24 hours) was treated with a compound of formula (I), formula (I)-A and formula (I)-B or a pharmaceutically acceptable salt thereof.

In another aspect, the present invention provides the use of a compound of formula (I), formula (I)-A and formula (I)-B, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicinal product.

In another aspect, the present invention provides the use of a compound of formula (I), formula (I)-A and formula (I)-B, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the prevention or treatment of a disorder or disease mediated by the mTOR pathway.

In another aspect, the present invention provides the use of a compound of formula (I), formula (I)-A and formula (I)-B, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the prevention or treatment of a disorder or disease selected from

- acute or chronic organ or tissue transplant rejection;

- graft-related vasculopathies;

- proliferation and migration of smooth muscle cells, leading to thickening of the vascular intima, blockage of blood vessels, obstructive coronary atherosclerosis, restenosis;

- autoimmune diseases and inflammatory conditions;

- diseases associated with the treatment and prevention of asthma;

- diseases associated with multiple drug resistance (MDR);

- infections caused by fungi;

- inflammation;

- infections;

- age-related diseases;

- neurodegenerative diseases;

- proliferative disorders, in particular cancer;

- epileptic seizures and disorders associated with epileptic seizures;

- mitochondrial myopathy and mitochondrial stress and

- treatable conditions that have been shown to increase the likelihood of age-related diseases, such as conditions in which there is an increase in the levels of cytokines that cause aging.

In another aspect, the present invention provides the use of a compound of formula (I), formula (I)-A and formula (I)-B, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the prevention or treatment of a disorder or disease that involves the process of fibrosis or inflammation.

In one embodiment, the disorder is selected from liver and kidney disorders.

In one embodiment, the liver disorder is selected from liver fibrosis, which occurs in end-stage liver disease; liver cirrhosis; liver failure caused by toxicity; non-alcoholic steatohepatitis or NASH and alcoholic steatosis.

In one embodiment, the kidney disorder is renal fibrosis, which occurs as a result of acute kidney injury.

In one embodiment, the kidney disorder is a chronic kidney disorder.

In one embodiment, the kidney disorder is diabetic nephropathy.

In another aspect, the present invention provides the use of a compound of formula (I), formula (I)-A and formula (I)-B, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the prevention or treatment of an age-related disorder or disease selected from sarcopenia , skin atrophy, types of senile hemangioma, types of seborrheic keratosis, brain atrophy, also called dementia, atherosclerosis, arteriosclerosis, emphysema, osteoporosis, osteoarthritis, high blood pressure, erectile dysfunction, types of cataracts, macular degeneration, glaucoma, stroke, cerebrovascular disease (stroke), chronic kidney disease, diabetes-associated kidney disease, liver dysfunction, liver fibrosis, autoimmune hepatitis, endometrial hyperplasia, metabolic disorder, renovascular disease, hearing loss, mobility impairment, cognitive decline, tendon stiffness, dysfunction heart disease, such as cardiac hypertrophy, and/or systolic and/or diastolic dysfunction, and/or hypertension, cardiac dysfunction leading to decreased ejection fraction, aging of the immune system, Parkinson's disease, Alzheimer's disease, cancer, aging of the immune system 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 diabetes-related complications such as kidney failure , blindness and neuropathy.

In another aspect, the present invention provides the use of a compound of formula (I), formula (I)-A and formula (I)-B, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the prevention or treatment of cancer.

In another aspect, the present invention provides the use of a compound of formula (I), formula (I)-A and formula (I)-B or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment of kidney cancer, renal cell carcinoma, colorectal cancer , uterine sarcoma, uterine endometrial cancer, endometrial cancer, breast cancer, ovarian cancer, cervical cancer, stomach cancer, fibrosarcoma, pancreatic cancer, liver cancer, melanoma, leukemia, multiple myeloma, nasopharyngeal cancer, prostate cancer, lung cancer , glioblastoma, bladder cancer, mesothelioma, head cancer, rhabdomyosarcoma, sarcoma, lymphoma or neck cancer.

Specific individual combinations that may provide specific treatment benefits include a combination of a compound

or a pharmaceutically acceptable salt thereof and an inhibitor of mTOR catalytic activity, in particular the inhibitor mentioned above. In one embodiment, the compound is (S)-C16-(1,1-dioxidoisothiazolidin-2-yl)-C32-deoxorapamycin (Example 1) or a pharmaceutically acceptable salt thereof. In one embodiment, the compound is (R)-C16-(1,1-dioxidoisothiazolidin-2-yl)-C32-deoxorapamycin (Example 2) or a pharmaceutically acceptable salt thereof.

In one embodiment, the present invention provides a product containing the compound

or a pharmaceutically acceptable salt thereof and an inhibitor of mTOR catalytic activity, in particular the inhibitor specified above, in the form of a combination preparation for simultaneous, separate or sequential use in therapy. In one embodiment, the compound is (S)-C16-(1,1-dioxidoisothiazolidin-2-yl)-C32-deoxorapamycin (Example 1) or a pharmaceutically acceptable salt thereof. In one embodiment, the compound is (R)-C16-(1,1-dioxidoisothiazolidin-2-yl)-C32-deoxorapamycin (Example 2) or a pharmaceutically acceptable salt thereof.

In one embodiment, the present invention provides a pharmaceutical composition containing the compound

or a pharmaceutically acceptable salt thereof and an inhibitor of mTOR catalytic activity, in particular the inhibitor set forth above, and a pharmaceutically acceptable carrier. In one embodiment, the compound is (S)-C16-(1,1-dioxidoisothiazolidin-2-yl)-C32-deoxorapamycin (Example 1) or a pharmaceutically acceptable salt thereof. In one embodiment, the compound is (R)-C16-(1,1-dioxidoisothiazolidin-2-yl)-C32-deoxorapamycin (Example 2) or a pharmaceutically acceptable salt thereof.

In one embodiment, the other therapeutic agent is selected from a CD4 lymphocyte depleting agent, such as an anti-CD4 antibody or an antigen-binding fragment thereof, such as a humanized anti-CD4 antibody, e.g. zanolimumab. Such a therapeutic agent can be used in particular for the treatment of a proliferative disorder, in particular cancer. See also US Patent No. 8906374 and US Patent No. 9427463 regarding such combination therapy.

Specific individual combinations that may provide specific treatment benefits include a combination of a compound

or a pharmaceutically acceptable salt thereof and a CD4 lymphocyte depleting agent, in particular the agent mentioned above. In one embodiment, the compound is (S)-C16-(1,1-dioxidoisothiazolidin-2-yl)-C32-deoxorapamycin (Example 1) or a pharmaceutically acceptable salt thereof. In one embodiment, the compound is (R)-C16-(1,1-dioxidoisothiazolidin-2-yl)-C32-deoxorapamycin (Example 2) or a pharmaceutically acceptable salt thereof.

In one embodiment, the present invention provides a product containing the compound

or a pharmaceutically acceptable salt thereof and a CD4 lymphocyte depleting agent, in particular the agent mentioned above, in the form of a combination preparation for simultaneous, separate or sequential use in therapy. In one embodiment, the compound is (S)-C16-(1,1-dioxidoisothiazolidin-2-yl)-C32-deoxorapamycin (Example 1) or a pharmaceutically acceptable salt thereof. In one embodiment, the compound is (R)-C16-(1,1-dioxidoisothiazolidin-2-yl)-C32-deoxorapamycin (Example 2) or a pharmaceutically acceptable salt thereof.

In one embodiment, the present invention provides a pharmaceutical composition containing the compound

or a pharmaceutically acceptable salt thereof, a CD4 lymphocyte depleting agent, in particular the agent set forth above, and a pharmaceutically acceptable carrier. In one embodiment, the compound is (S)-C16-(1,1-dioxidoisothiazolidin-2-yl)-C32-deoxorapamycin (Example 1) or a pharmaceutically acceptable salt thereof. In one embodiment, the compound is (R)-C16-(1,1-dioxidoisothiazolidin-2-yl)-C32-deoxorapamycin (Example 2) or a pharmaceutically acceptable salt thereof.

In one embodiment, the other therapeutic agent is selected from an agent that modulates the activity of immunoinhibitory proteins such as PD-1/PDL-1, for example, an anti-PD-1 antibody or a PDL-1 antibody. Anti-PD-1 antibodies that are useful as such a therapeutic agent are disclosed in US Pat. No. 9,683,048. Such a therapeutic agent can be used in the treatment of cancer, in particular for cancer immunotherapy.

Specific individual combinations that may provide specific treatment benefits include a combination of a compound

or a pharmaceutically acceptable salt thereof and an agent that modulates the activity of immunoinhibitory proteins such as PD-1, in particular the agent mentioned above. In one embodiment, the compound is (S)-C16-(1,1-dioxidoisothiazolidin-2-yl)-C32-deoxorapamycin (Example 1) or a pharmaceutically acceptable salt thereof. In one embodiment, the compound is (R)-C16-(1,1-dioxidoisothiazolidin-2-yl)-C32-deoxorapamycin (Example 2) or a pharmaceutically acceptable salt thereof.

In one embodiment, the present invention provides a product containing the compound

or a pharmaceutically acceptable salt thereof and an agent that modulates the activity of immunoinhibitory proteins such as PD-1, in particular the agent mentioned above, in the form of a combination preparation for simultaneous, separate or sequential use in therapy. In one embodiment, the compound is (S)-C16-(1,1-dioxidoisothiazolidin-2-yl)-C32-deoxorapamycin (Example 1) or a pharmaceutically acceptable salt thereof. In one embodiment, the compound is (R)-C16-(1,1-dioxidoisothiazolidin-2-yl)-C32-deoxorapamycin (Example 2) or a pharmaceutically acceptable salt thereof.

In one embodiment, the present invention provides a pharmaceutical composition containing

or a pharmaceutically acceptable salt thereof, an agent that modulates the activity of immunoinhibitory proteins such as PD-1, in particular an agent as defined above, and a pharmaceutically acceptable carrier. In one embodiment, the compound is (S)-C16-(1,1-dioxidoisothiazolidin-2-yl)-C32-deoxorapamycin (Example 1) or a pharmaceutically acceptable salt thereof. In one embodiment, the compound is (R)-C16-(1,1-dioxidoisothiazolidin-2-yl)-C32-deoxorapamycin (Example 2) or a pharmaceutically acceptable salt thereof.

EXAMPLES

The present invention provides the following examples. The synthetic examples and biological examples described in this application are presented to illustrate the compounds, pharmaceutical compositions and methods provided herein and should not be construed as limiting their scope in any way.

The compounds provided herein can be prepared from readily available starting materials using modifications to the specific synthetic protocols set forth below, as will be generally known to those skilled in the art. It will be understood that if typical or preferred process conditions are indicated (i.e. reaction temperature values, time values, molar ratios of reagents, solvents, pressure values and etc.), other method conditions can also be used unless otherwise indicated. Optimal reaction conditions may vary depending on the particular reagents or solvents used, but such conditions can be determined by those skilled in the art through routine optimization procedures.

In addition, those skilled in the art will appreciate that the use of traditional protecting groups may be necessary to prevent unwanted reactions with certain functional groups. The selection of a suitable protecting group for a particular functional group, as well as suitable conditions for protecting and deprotecting the group, are well known in the art. For example, numerous protecting groups, and their introduction and removal, are described in Greene et al. , Protecting Groups in Organic Synthesis , Second Edition, Wiley, New York, 1991, and references cited therein.

Rapamycin and its derivatives, for example, the compounds of formula (I), formula (I)-A and formula (I)-B, exist as a solution-dependent and pH-dependent equilibrium of six-membered and seven-membered hemiketal forms, shown below as E and F ( schemes 4 and 5 ). See The Journal of Antibiotics (Tokyo) (1991) 44(6):688-90 and Tetrahedron Letters (1992) 33(33):4139-4142. Rapamycin and its derivatives also exist as a mixture of cis and trans amides, shown below as E , H , J and K ( Schemes 4 and 5 ). [Cm. Mierke, D. F., Schmieder, P., Karuso, P. and Kessler, H. (1991), Conformational Analysis of the cis and trans -Isomers of FK506 by NMR and Molecular Dynamics. Helvetica Chimica Acta , 74: 1027-1047.] The NMR characterization data shown in the examples correspond only to the major equilibrium form observed under the indicated conditions in deuterated solvent.

Scheme 4

Where

R ₁ is selected from the group consisting of hydroxy,

R ₂ is selected from the group consisting of

, Where

m is 0, 1, 2 or 3;

n is 1, 2 or 3;

o equals 1, 2, 3, 4, 5 or 6;

p equals 1, 2, 3, 4 or 5;

q is equal to 1, 2, 3, 4 or 5, with the sum of p and q being 2, 3, 4, 5 or 6;

r equals 2, 3 or 4;

s is equal to 2, 3 or 4, with the sum of r and s being 4, 5 or 6;

X is O, S, NR ₆ or SO ₂ ;

R ₃ represents hydrogen, C _1-6 alkyl, hydroxy-C _1-6 alkyl, C _3-8 cycloalkyl-C _0-6 alkyl or phenyl-C _0-6 alkyl;

R ₄ is hydrogen and R ₅ is hydrogen, hydroxy or cyano, or R ₄ and R ₅ together form =O; And

R ₆ is hydrogen, C _1-6 alkyl, C _3-8 cycloalkyl-C _0-6 alkyl, phenyl-C _0-6 alkyl, C _1-6 alkyl-CO-, C _3-8 cycloalkyl-C _{0- 6} alkyl-CO-, C _1-6 alkyl-SO ₂ - or C _3-8 cycloalkyl-C _0-6 alkyl-SO ₂ -.

In one embodiment, R ₁ is hydroxy.

In one embodiment, R ₂ is

, and n is 1, 2 or 3. In one embodiment, R ₂ is .

Scheme 5

Where

R ₁ represents , And

R ₂ is selected from the group consisting of

, Where

m is 0, 1, 2 or 3;

n is 1, 2 or 3;

o equals 1, 2, 3, 4, 5 or 6;

p equals 1, 2, 3, 4 or 5;

q is equal to 1, 2, 3, 4 or 5, with the sum of p and q being 2, 3, 4, 5 or 6;

r equals 2, 3 or 4;

s is equal to 2, 3 or 4, with the sum of r and s being 4, 5 or 6;

X is O, S, NR ₆ or SO ₂ ;

R ₃ represents hydrogen, C _1-6 alkyl, hydroxy-C _1-6 alkyl, C _3-8 cycloalkyl-C _0-6 alkyl or phenyl-C _0-6 alkyl;

R ₄ is hydrogen and R ₅ is hydrogen, hydroxy or cyano, or R ₄ and R ₅ together form =O; And

R ₆ is hydrogen, C _1-6 alkyl, C _3-8 cycloalkyl-C _0-6 alkyl, phenyl-C _0-6 alkyl, C _1-6 alkyl-CO-, C _3-8 cycloalkyl-C _{0- 6} alkyl-CO-, C _1-6 alkyl-SO ₂ - or C _3-8 cycloalkyl-C _0-6 alkyl-SO ₂ -.

In one embodiment, R ₂ is

, and n is 1, 2 or 3. In one embodiment, R ₂ is .

In one embodiment, the compounds of formula (I), formula (I)-A and formula (I)-B exist as a solvent-dependent and pH-dependent equilibrium of six-membered and seven-membered hemiketal forms, shown below as E-1 and F-1 ( Scheme 6 ). In one embodiment, the compounds of formula (I), formula (I)-A and formula (I)-B exist as a mixture of cis and trans amides E-1 and H-1 .

Scheme 6

Receiving connections

The compounds of the present invention can be prepared as described in the following examples.

Abbreviations

4-EP 4-ethylpyridine

AcOH acetic acid

ACN Acetonitrile

aq. water

C degrees Celsius

d doublet

dd doublet of doublets

DCM dichloromethane

DIPEA N, N -diisopropylethylamine

DMSO dimethyl sulfoxide

EDTA ethylenediaminetetraacetic acid

ESIMS electrospray ionization mass spectrometry

EtOAc ethyl acetate

EtOH ethanol

FA formic acid

g gram

h. hour(hours)

HPLC high performance liquid chromatography

HRMS High Resolution Mass Spectrometry

iPrOH 2-propanol or isopropanol

l liter

LC liquid chromatography

LCMS liquid chromatography and mass spectrometry

MeOH methanol

MS mass spectrometry

M mol/l

m multiplet

min. minutes

ml milliliter(milliliters)

µM µmol/l

mass/charge ratio of mass and charge

N ₂ nitrogen gas

nM nmol/l

NMR nuclear magnetic resonance

¹ H NMR: Proton Nuclear Magnetic Resonance Spectrometry

PEI polyethylenimine

PPU propylpyridyl urea

pTsOH p-toluenesulfonic acid

Rev. preparative

rac racemic

rpm rpm

room temperature

s singlet

sat. saturated

SFC supercritical fluid chromatography

t triplet

TCEP tris(2-carboxyethyl)phosphine

TFA trifluoroacetic acid

THF tetrahydrofuran

TLC thin layer chromatography

about. volume

Methods Used to Purify Example Compounds

Purification of intermediates and final products was accomplished using either normal phase or reverse phase chromatography.

Normal phase chromatography was performed using pre-filled SiO cartridges₂ (For example, RediSep® Rf columns from Teledyne Isco, Inc.) with elution gradients of suitable solvent systems (e.g. hexanes and ethyl acetate; DCM and MeOH; or unless otherwise stated).

SFC was performed using the methods described below.

Method 1: Princeton PPU 5 µm (100A) column (30 × 250 mm); _CO2 /MeOH

Method 2: Princeton 4-EP 5 µm (60A) column (30 × 250 mm); _CO2 /MeOH

Method 3: Reprospher PEI 5 µm column (100A) (30 × 250 mm); _CO2 /MeOH

Gradients were chosen based on analytical separation.

Preparative reverse phase HPLC was performed using the methods described below.

Method 1: column (Agilent) Phenomenex Luna C18; 5 µm (30 × 250 mm); 0.1% formic acid and 5% water in acetonitrile; 0.1% formic acid and 5% acetonitrile in water. Gradients were chosen based on analytical separation.

Method 2: column (EZprep) YMC Actus Triart C18; 5 µm (20 × 150 mm); acetonitrile/water. Gradients were chosen based on analytical separation.

Chiral preparative HPLC was performed using the methods described below.

Method 1: Chiralpak IC column; 5 µm (20 ×250 mm); n-heptane/DCM/EtOH

Method 2: Chiralpak ID column; 5 µm (20 × 250 mm); n-heptane/DCM/iPrOH

Gradients were chosen based on analytical separation.

LC/MS method

Column: Acquity UPLC BEH C18, 130 angstrom, 1.7 µm, 2.1 mm × 50 mm

temperature: 50°C

Administration: 1 µl

Solvent A: water+5 mM ammonium hydroxide

Solvent B: acetonitrile + 5 mM ammonium hydroxide

Gradient:

Time (min.) Consumption A% B% Curve Original value 1,000 98.0 2.0 Original value 4.40 1.0 2.0 98.0 6 5.15 1.0 2.0 98.0 6 5.19 1.0 98.0 2.0 6

Equipment for ¹ H NMR:

Bruker UltraShield™ Advance III HD operating at 400 MHz with cryo-DCI probe. Data were processed using MestReNova 11.0 software.

Preparation of intermediates 1-7

Intermediate 1: C32-deoxorapamycin

Intermediate 1 was prepared according to procedures known in the literature, including those disclosed in US Patent Publication No. 005985890 and WO 2007/085400 A1, each of which is incorporated by reference herein in its entirety.

Intermediate 2: RAD001(everolimus; Afinitor®)

Intermediate 2

Intermediate 2 was prepared according to procedures known in the literature, including those disclosed in WO 2012103959, which is incorporated herein by reference in its entirety.

Intermediate 3:

Intermediate 3 was prepared in two steps from Intermediate A as shown below.

Step 1: Synthesis of Intermediate A

Dissolve 2-((tert-butyldimethylsilyl)oxy)ethanol (0.471 g, 2.67 mmol) in anhydrous toluene (0.95 ml) in a reaction vessel. The vessel was sealed and then evacuated using nitrogen. AddedN, N-diisopropylethylamine (DIPEA) (0.490 ml, 2.81 mmol) using a syringe. The mixture was cooled to 0°C in an ice bath. Triflate anhydride (Tf) was added dropwise₂O) (0.438 ml, 2.59 mmol) at 0°C for a period of approximately two minutes. The reaction mixture was stirred at 0°C for 30 minutes.

The vessel was removed from the cooling bath. DIPEA (0.490 mL, 2.81 mmol) was added via syringe. The vessel was opened and Intermediate 1 (0.600 g, 0.667 mmol) was quickly added in one portion. The vessel was quickly closed again and the mixture was evacuated using nitrogen. Toluene (0.5 ml) was added.

The reaction mixture was stirred at 40°C under nitrogen atmosphere overnight. The reaction mixture was diluted with saturated aqueous NaHCO ₃ solution. The slaked mixture was extracted five times with EtOAc. The organic extracts were combined, dried over Na ₂ SO ₄ , vacuum filtered through celite and concentrated to give a waxy white crude solid.

The crude product was purified by silica gel flash column chromatography (0 to 35% acetone in heptane elution gradient, 40 g silica column, TLC in 35% acetone in heptane, UV visualized) to give the desired intermediate A (0.245 g, 0.231 mmol, 34.7% yield) as a glassy substance, which was directly used as is in the next step.

Intermediate A : ESIMS [M+NH4] 1076.1, ESIMS [MH] 1056.0.

Step 2. Synthesis of intermediate 3

Dissolve intermediate A (0.135 g, 0.128 mmol) in anhydrous THF (1.2 ml) in a glass reaction vessel. The vessel was sealed and the mixture was evacuated twice using nitrogen. The mixture was cooled to 0°C in an ice bath. HF-pyridine (0.12 mL, 1.332 mmol) was added dropwise using a syringe over a period of 30 seconds. The reaction mixture was stirred at 0°C for 60 minutes.

The reaction mixture was added dropwise to a saturated aqueous NaHCO ₃ solution. The slaked mixture was extracted five times with EtOAc. The organic extracts were combined, dried over Na ₂ SO ₄ , vacuum filtered through celite and concentrated to give a white solid crude product.

The crude product was purified by silica gel flash column chromatography (0 to 50% acetone in heptane elution gradient, 40 g silica column, TLC in 50% acetone in heptane, UV visualized) to give intermediate 3 (0.087 g , 0.092 mmol, yield 72.2%) as a white solid.

Intermediate 3: ESIMS [M+NH4] 962.0, ESIMS [MH] 943.0.

^1H NMR (400 MHz, Chloroform- d ) δ 6.46-6.25 (m, 2H), 6.19-6.09 (m, 1H), 5.91 (m, 1H), 5.55 (m, 1H), 5.34-5.26 (m, 1H), 5.26-5.15 (m, 1H), 4.88-4.64 (m, 2H), 4.11 (m , 1H), 3.94-3.82 (m, 1H), 3.79 (m, 1H), 3.73-3.63 (m, 3H), 3.63-3.47 (m, 4H ), 3.45 (m, 4H), 3.33 (m, 3H), 3.28-3.15 (m, 2H), 3.13 (s, 3H), 3.08 (m, 1H) , 2.80 (m, 1H), 2.32 (m, 3H), 2.14 (m, 2H), 2.07-1.94 (m, 2H), 1.94-1.72 (m , 5H), 1.72-1.56 (m, 8H), 1.56-1.40 (m, 3H), 1.39-1.21 (m, 7H), 1.21-1.09 (m, 1H), 1.08-0.99 (m, 8H), 0.98-0.83 (m, 9H), 0.73 (q, J =12.0 Hz, 1H).

Intermediate 4:

Combine intermediate 1 (0.233 g, 0.259 mmol) with 2,6-di-tert-butyl-4-methylpyridine (0.425 g, 2.071 mmol) in anhydrous dichloromethane (2.6 ml). The reaction mixture was evacuated once with nitrogen. The reaction mixture was cooled to 0°C in an ice bath. Solid dimethylphosphine chloride (0.146 g, 1.294 mmol) was added in one portion. The reaction mixture was stirred at 0°C for 80 minutes.

The reaction mixture was diluted with saturated aqueous NaHCO3 and extracted several times with EtOAc. The organic extracts were combined, dried over Na ₂ SO ₄ , decanted and concentrated to give the crude product as a colorless gum (0.768 g).

The crude product was purified by silica gel flash column chromatography (0 to 80% acetone in heptane elution gradient, 24 g silica column, TLC in 80% EtOAc in heptane, visualized under UV). Fractions containing product were combined and concentrated to give intermediate 4 (0.087 g, 0.089 mmol, 34.4% yield) as a white solid.

Intermediate 4: ESIMS [M+NH4] 993.7, ESIMS [MH] 974.7.

HRMS: Calculated: 999.5812 (as sodium adduct). Discovered: 999.5807.

^1H NMR (600 MHz, Chloroform- d ) δ 6.47-6.26 (m, 2H), 6.22-6.08 (m, 1H), 6.02-5.83 (m, 1H) , 5.54 (m, 1H), 5.35-5.26 (m, 1H), 5.21 (m, 1H), 4.85-4.76 (m, 1H), 4.12 (m , 2H), 3.93-3.81 (m, 1H), 3.67 (t, J =7.7 Hz, 1H), 3.62 (d, J =6.7 Hz, 1H), 3 ,60-3.53 (m, 1H), 3.53-3.44 (m, 1H), 3.42-3.36 (m, 3H), 3.32 (m, 3H), 3.28 -3.18 (m, 1H), 3.13 (m, 3H), 3.05 (m, 1H), 2.82 (m, 1H), 2.42-2.21 (m, 3H), 2.16-2.08 (m, 3H), 1.99 (m, 1H), 1.95-1.83 (m, 1H), 1.83-1.72 (m, 4H), 1. 71-1.57 (m, 9H), 1.57-1.43 (m, 12H), 1.39 (m, 1H), 1.34-1.20 (m, 4H), 1.20- 1.10 (m, 1H), 1.05 (m, 4H), 1.00 (d, J =6.5 Hz, 3H), 0.95 (dd, J =6.6, 2.1 Hz , 3H), 0.92 (d, J =6.6 Hz, 3H), 0.91-0.84 (m, 4H), 0.77 (q, J =12.1 Hz, 1H).

Intermediate 5:

Dissolve intermediate 1 (4.372 g, 4.86 mmol) in anhydrous dichloromethane (20 ml). Anhydrous toluene (20 ml) was added. The reaction mixture was evaporated to dryness on a rotary evaporator. This azeotropic drying process was repeated two more times.

The dried starting material was combined with 2,6-lutidine (1.388 ml, 11.91 mmol) in anhydrous dichloromethane (58 ml). The flask was capped and the mixture was evacuated twice with nitrogen. The mixture was cooled to -30°C in an acetonitrile/dry ice bath.

Triflate anhydride (1.209 mL, 7.16 mmol) was added dropwise via syringe over a period of four minutes. The reaction mixture was stirred at -30°C for 30 minutes. The reaction mixture was transferred to a 0°C ice bath and stirred for 20 minutes at 0°C.

The reaction mixture was placed on a rotary evaporator and concentrated without heating. Isopropyl acetate (22 ml) was added. Tetrazole (1.170 g, 16.71 mmol) was added in one portion. The flask was quickly capped and evacuated twice using nitrogen. N,N -diisopropylethylamine (4.18 mL, 23.94 mmol) was added via syringe over a period of one minute. The reaction mixture was stirred overnight at room temperature.

The reaction mixture was concentrated on a rotary evaporator. The concentrate was purified by normal phase silica gel flash column chromatography (0 to 40% acetone in heptane elution gradient, 80 g silica column, TLC 40% acetone in heptane, visualized under UV).

Fractions corresponding to the second elution peak (determined by UV absorbance at 279 nm) were combined and concentrated to give intermediate 5 (2.194 g, 2.304 mmol, 47.4% yield) as a white solid.

Intermediate 5: ESIMS [M+NH4] 969.8, ESIMS [MH] 950.8.

^1H NMR (400 MHz, DMSO- d6 ) δ 9.33 (d, J =6.4 Hz, 1H) _, 6.58-6.41 (m, 2H), 6.35-6.14 ( m, 2H), 6.09-5.98 (m, 1H), 5.55-5.43 (m, 1H), 5.19 (m, 1H), 5.05 (m, 1H), 5 .00-4.93 (m, 1H), 4.87-4.79 (m, 1H), 4.67-4.56 (m, 1H), 3.98-3.87 (m, 1H) , 3.87 (d, J =6.9 Hz, 1H), 3.61 (m, 2H), 3.55 (dd, J =11.8, 1.9 Hz, 1H), 3.49- 3.38 (m, 1H), 3.31-3.17 (m, 4H), 3.10 (m, 4H), 3.04 (s, 3H), 2.88-2.75 (m, 1H), 2.29-2.09 (m, 3H), 2.07-1.86 (m, 3H), 1.88-1.60 (m, 9H), 1.59-1.44 ( m, 7H), 1.43-1.01 (m, 11H), 0.96 (t, J =7.1 Hz, 5H), 0.95-0.77 (m, 7H), 0.72 (m, 4H).

Intermediate 6:

Combine 2-ethoxyethanol (0.538 mL, 5.55 mmol) in anhydrous toluene (2.0 mL) and anhydrous dioxane (0.22 mL). N,N -diisopropylethylamine (1.067 mL, 6.11 mmol) was added via syringe. The reaction mixture was evacuated twice with nitrogen. The mixture was cooled to 0°C in an ice bath.

Triflate anhydride (0.901 mL, 5.33 mmol) was added dropwise using a syringe over a period of two minutes. The reaction mixture was stirred at 0°C for 15 minutes. The cooling bath was removed and the reaction mixture was allowed to equilibrate at room temperature for 15 minutes.

N,N -diisopropylethylamine (1.067 mL, 6.11 mmol) was added dropwise via syringe over a period of 30 seconds. Intermediate 1 (1.00 g, 1.111 mmol) was added in one portion. The reaction mixture was quickly capped and evacuated with nitrogen. Anhydrous toluene (2.0 ml) and anhydrous dioxane (0.22 ml) were added using a syringe. The mixture was stirred at 55°C for 24 hours.

The reaction mixture was diluted with brine and extracted several times with EtOAc. The organic extracts were combined, dried over Na ₂ SO ₄ , decanted and concentrated to give the crude product as a colorless gum (4.25 g).

The crude product was purified by silica gel flash column chromatography (0 to 50% acetone in heptane elution gradient, 80 g silica column, TLC in 40% acetone in heptane, visualized under UV). Fractions containing product were combined and concentrated to give intermediate 6 (0.443 g, 0.456 mmol, 41.0% yield) as a colorless glass.

Intermediate 6: ESIMS [MH] 970.9.

HRMS: calculated - 989.6678 (as ammonium adduct); detected - 989.6655

Calculated - 994.6232 (as an adduct with sodium); detected - 994.6215.

Intermediate 7:

Intermediate 7 was prepared in two steps from Intermediate B as shown below.

Step 1: Synthesis of Intermediate B

Combine 2,2,5-trimethyl-1,3-dioxane-5-carboxylic acid (0.400 g, 2.296 mmol) and triethylamine (0.320 mL, 2.296 mmol) and dissolve in anhydrous THF (7.6 mL). The reaction mixture was cooled to 0°C. 2,4,6-trichlorobenzoyl chloride (0.359 mL, 2.296 mmol) was added via syringe. The reaction mixture was stirred at 0°C for ten minutes. The cooling bath was removed and the reaction mixture was stirred at room temperature for 2.5 hours. A white solid precipitated.

The reaction mixture was filtered through a syringe filter. The filter was rinsed with THF (2 ml). The filtrates were combined and concentrated on a rotary evaporator to obtain a colorless gum.

Toluene (7.6 ml) was added to the concentrate. Intermediate 1 (1.447 g, 1.607 mmol) was added in one portion, followed by 4-dimethylaminopyridine (0.281 g, 2.296 mmol). The reaction mixture was stirred at room temperature overnight.

The mixture was diluted with water and extracted four times with EtOAc. The organic extracts were combined, dried over Na ₂ SO ₄ , decanted and concentrated to give the crude product as a yellow gum (2.15 g).

The crude product was purified by silica gel flash column chromatography (0 to 40% acetone in heptane elution gradient, 40 g silica column, TLC in 40% acetone in heptane, visualized under UV). Fractions containing product were combined and concentrated to give Intermediate B (0.438 g, 0.415 mmol, 18.1% yield) as a white foam.

Intermediate B : ESIMS [M+NH4] 1074.0, ESIMS [MH] 1055.1.

Step 2. Synthesis of intermediate 7

Dissolve intermediate B (0.431 g, 0.408 mmol) in THF (4 ml). HCl (1 M eq) (2.0 mL, 2.00 mmol) was added via syringe. The reaction mixture was stirred at room temperature for 36 hours.

The reaction mixture was diluted with water and extracted four times with EtOAc. The organic extracts were combined, dried over Na ₂ SO ₄ , decanted and concentrated to give a pale yellow resin. The resin was dissolved in dichloromethane and diluted with heptane. The mixture was concentrated and dried under high vacuum to give intermediate 7 (0.407 g, 0.400 mmol, 98% yield) as a white solid.

Intermediate 7 : ESIMS [M+NH4] 1034.0, ESIMS [MH] 1015.0.

HRMS: calculated - 1033.6576 (as ammonium adduct); detected - 1033.6588

Calculated - 1038.6130 (as an adduct with sodium); discovered - 1038.6138.

Example 1: ( S )-C16-(1,1-dioxidoizothiazolidin-2-yl)-C32-deoxyrapamycin

Example 2: ( R )-C16-(1,1-dioxidoizothiazolidin-2-yl)-C32-deoxyrapamycin

To a stirred solution of intermediate 1 (2.0 g, 2.222 mmol) and isothiazolidine-1,1-dioxide (4.04 g, 33.3 mmol, 15 equivalents) in anhydrous dichloromethane (5.6 mL) was added one portion -toluenesulfonic acid⋅H ₂ O (0.042 g, 0.222 mmol, 0.1 equivalent). The reaction mixture was stirred at ambient temperature under nitrogen atmosphere for 34 minutes. The entire reaction mixture was directly chromatographed on silica (eluting gradient from 100% heptane to 40% acetone in heptane) to give a product that was a mixture of both diastereomers in a ratio of approximately 3:1, determined by UV absorbance at 279 nm in the assay LC/MS.

The diastereomeric mixture was separated by normal phase chromatography on silica (elution gradient from 100% dichloromethane to 40% acetonitrile in dichloromethane).

The first diastereomer eluted (Rf ~0.23 on silica in TLC with 30% acetonitrile in dichloromethane) gave the (S)-diastereomer of Example 1 as a white solid.

Example 1: ESIMS [M+NH4] 1006.7, ESIMS [MH] 987.8.

¹H NMR (600 MHz, Chloroform-d) δ 6.43 (dd,J=14.9, 10.4 Hz, 1H), 6.35 (dd,J=14.9, 10.7 Hz, 1H), 6.16 (dd,J=15.1, 10.2 Hz, 1H), 6.06-6.01 (m, 1H), 5.67 (dd,J=15.2, 8.6 Hz, 1H), 5.35 (dd,J=6.4, 1.8 Hz, 1H), 5.26 (d,J=9.6 Hz, 1H), 4.83 (td,J=6.6, 4.7 Hz, 1H), 4.12 (d,J=7.4 Hz, 1H), 3.88 (dd,J=11.1, 5.1 Hz, 1H), 3.84-3.77 (m, 1H), 3.63-3.60 (m, 2H), 3.51 (d,J=7.5 Hz, 1H), 3.46 (s, 3H), 3.45-3.42 (m, 3H), 3.35 (s, 3H), 3.29-3.14 (m, 3H), 3.06-2.96 (m, 2H), 2.93 (ddd,J=10.4, 6.4, 1.5 Hz, 1H), 2.44 (tt,J=8.6, 6.2 Hz, 1H), 2.37-2.23 (m, 4H), 2.22-2.14 (m, 2H), 2.03 (dt,J=12.3, 3.8 Hz, 1H), 1.96 (pd,J=6.4, 5.7, 3.5 Hz, 2H), 1.92-1.82 (m, 3H), 1.80-1.76 (m, 2H), 1.75 (d,J=1.2 Hz, 4H), 1.72 (d,J=3.1 Hz, 1H), 1.68 (d,J=1.3 Hz, 3H), 1.65-1.53 (m, 4H), 1.48-1.17 (m, 9H), 1.07 (d,J=6.5 Hz, 1H), 1.06 (s, 3H), 1.04 (d,J=7.3 Hz, 3H), 1.01 (d,J=6.6 Hz, 3H), 0.99 (dd,J=6.7, 2.3 Hz, 3H), 0.95 (d,J=6.8 Hz, 3H), 0.74 (q,J=11.9 Hz, 1H).

The second diastereomer eluted (Rf ~0.16 on silica in TLC with 30% acetonitrile in dichloromethane) gave the (R)-diastereomer of Example 2 as a white solid.

Example 2 : ESIMS [M+NH4] 1006.9, ESIMS [MH] 988.1.

^1H NMR (Chloroform- d ) δ 6.48 (dd, J=14.7, 10.9 Hz, 1H), 6.24 (dd, J=14.6, 10.6 Hz, 1H), 6 .16 (dd, J=14.9, 10.6 Hz, 1H), 6.01 (d, J=11.0 Hz, 1H), 5.38 (dd, J=14.9, 9.8 Hz, 1H), 5.23 (dd, J=6.2, 2.0 Hz, 1H), 5.12 (d, J=9.9 Hz, 1H), 4.73 (dd, J=12 ,1, 2.9 Hz, 1H), 4.65 (dt, J=8.3, 3.9 Hz, 1H), 4.14 (d, J=6.7 Hz, 1H), 3.97 (m, 1H), 3.74 (d, J=6.7 Hz, 1H), 3.62 (qd, J=13.9, 12.7, 5.5 Hz, 2H), 3.42 ( s, 3H), 3.39 (m, 1H), 3.31 (s, 3H), 3.24 (ddd, J=12.2, 7.5, 4.6 Hz, 1H), 3.10 (td, J=8.2, 3.8 Hz, 1H), 3.08 (s, 1H), 3.02-2.97 (m, 1H), 2.97-2.92 (m, 1H ), 2.83-2.71 (m, 1H), 2.42 (ddt, J=13.1, 9.5, 6.4 Hz, 1H), 2.34 (d, J=4.3 Hz, 1H), 2.31 (s, 1H), 2.28-2.23 (m, 1H), 2.22-2.19 (m, 1H), 2.19 (s, 2H), 2 .12-2.08 (m, 2H), 2.03-1.99 (m, 1H), 1.90 (s, 3H), 1.88 (s, 1H), 1.79 (s, 1H ), 1.77 (s, 1H), 1.76 (s, 1H), 1.72-1.68 (m, 1H), 1.48 (s, 1H), 1.46 (s, 1H) , 1.40 (d, J=3.0 Hz, 1H), 1.38 (s, 1H), 1.66 (d, J=3.0 Hz, 2H), 1.64 (d, J= 2.9 Hz, 1H), 1.62 (s, 2H), 1.62 (s, 2H), 1.58-1.53 (m, 1H), 1.37 (s, 1H), 1, 36 (d, J=2.3 Hz, 1H), 1.33 (d, J=2.9 Hz, 1H), 1.30 (dd, J=6.7, 1.8 Hz, 1H), 1.28 (s, 2H), 1.28 (s, 2H), 1.24 (s, 1H), 1.09 (s, 1H), 1.07 (d, J=6.6 Hz, 3H ), 1.05 (d, J=6.6 Hz, 3H), 1.01 (d, J=3.2 Hz, 1H), 0.95 (d, J=6.8 Hz, 3H), 0.92 (s, 1H), 0.92-0.90 (m, 3H), 0.88 (d, J=6.8 Hz, 2H), 0.66 (q, J=12.0 Hz , 1H)

The absolute configurations of the C16 substituents in Example 1 and Example 2 were determined by X-ray crystallography when co-crystallized with FKBP12. [Cm. Stuart L. Schrieber and Jon Clardy, et al., Atomic Structure of the Rapamycin Humano Immunophilin FKBP-12 Complex, J. Am. Chem. Soc. , 1991 , 113 , 7433-7434.] Crystal structures are depicted in Figures 1A and 1B and Figures 2A and 2B .

Pure FKBP12(1-108) protein was concentrated to 9 mg/ml in 50 mM Tris pH 8.0, 150 mM NaCl, 1 mM EDTA, 1 mM TCEP. The co-crystallization complex was prepared by mixing the protein with 3 mM compound (from a 50 mM stock solution prepared in 90% dDMSO, 10% D2O). The complex was incubated for two hours at 4°C, then centrifuged at 10,000 rpm. for 2 minutes to remove any possible sediment before crystallization. Cocrystals were prepared at 20°C and by sessile drop vapor diffusion using crystal microtransfer screening [Allan D'Arcy et al. , An automated microseed matrix-screening method for protein crystallization, Acta Cryst. , (2007) D 63 , 550-554.] The droplets consisted of 200 nl of protein solution, 160 nl of reservoir solution and 40 nl of initial seed. Crystals appeared within a few days in condition A1 of the commercially available Qiagen ammonium sulfate screening kit. The reservoir solution consisted of 2.2 M ammonium sulfate. The crystals were protected from low temperature in a reservoir solution supplemented with 20% ethylene glycol and flash frozen in liquid nitrogen. Data were collected at the Swiss Light Source facility (SLS, Villigen, Switzerland) at the X10SA synchrotron radiation source experimental station.

Data were processed using XDS (Kabsch, W. (2010), XDS. Acta Cryst. D, 66: 125-132). Structures were determined by molecular replacement (Collaborative Computational Project, Number 4 (1994). Acta Cryst. D50, 760-763) using previous X-ray structures of FKBP12 as a search model. We used REFMAC programs (Murshudov GN, Lebedev A.A., et al., REFMAC 5 for the refinement of macromolecular crystal structures. Acta Crystallographica Section D: Biological Crystallography . 2011; 67 (Pt 4): 355-367) and COOT (Emsley P, Lohkamp B, Scott WG, Cowtan K. Features and development of Coot , Acta Crystallographica Section D: Biological Crystallography . 2010; 66 (Pt 4): 486-501 ) to clarify the structure and (re)build the model.

In the following examples, the absolute stereochemical configuration of the C16 substituent has not been determined by cocrystallization x-ray diffraction and is not known. In some examples, only the major diastereomer of the reaction product was isolated and its characteristics were described. In other examples, each diastereomer was isolated and its characteristics were described without assigning absolute stereochemistry.

Example 3: C16-(4-oxoazetidin-2-yl)-C32-deoxyrapamycin

Example 3

* Absolute stereochemical configuration for C16 is not specified

To a solution of intermediate 1 (130 mg, 0.144 mmol, 1.0 eq) and azetidin-2-one (205 mg, 2.89 mmol, 20 eq) in acetonitrile (3 ml) was added p -toluenesulfonic acid monohydrate ( 82 mg, 0.433 mmol, 3 eq.). The reaction mixture was stirred at room temperature for 15 minutes. and then diluted with H ₂ O and extracted with ethyl acetate. The organic extract was evaporated under reduced pressure. The crude product was purified by preparative HPLC chromatography (Method 1) followed by purification by SFC (Method 1) to give Example 3 (2.5 mg, 1.7% yield) as a white solid.

Example 3 : ESIMS [MH] 938.0.

Exact weight: 938.59

^1H NMR (400 MHz, DMSO- d6 ) δ 6.45 (dd, J=14.0, _11.0 Hz, 1H), 6.28-6.11 (m, 2H), 5.98 ( d, J=10.9 Hz, 1H), 5.95-5.85 (m, 1H), 5.50 (dd, J=14.2, 9.5 Hz, 1H), 5.06 (d , J=4.7 Hz, 1H), 5.02-4.97 (m, 1H), 4.94 (d, J=9.8 Hz, 1H), 4.63-4.53 (m, 2H), 4.34-4.22 (m, 1H), 3.99-3.92 (m, 1H), 3.84-3.74 (m, 1H), 3.61-3.53 ( m, 1H), 3.49 (d, J=7.3 Hz, 1H), 3.46-3.39 (m, 1H), 3.37-3.25 (m, 3H), 3.22 -3.12 (m, 4H), 3.10-3.04 (m, 1H), 3.04-2.95 (m, 1H), 2.92-2.65 (m, 4H), 2 .27-2.11 (m, 2H), 2.10-1.99 (m, 2H), 1.96-1.87 (m, 1H), 1.84-1.70 (m, 5H) , 1.69-1.37 (m, 15H), 1.37-1.05 (m, 8H), 1.03-0.93 (m, 4H), 0.90 (d, J=6, 4 Hz, 3H), 0.88-0.83 (m, 5H), 0.79 (d, J=6.7 Hz, 3H), 0.75 (d, J=6.7 Hz, 3H) , 0.59 (q, J=11.8 Hz, 1H).

Example 4: C16-(3-methylimidazolidin-2-on-1-yl)-C32-deoxyrapamycin

Example 4

* Absolute stereochemical configuration for C16 is not specified

To a solution of intermediate 1 (100 mg, 0.111 mmol, 1.0 eq.) and 1-methylimidazolidin-2-one (167 mg, 1.66 mmol, 15 eq.) in DCM (2 mL) was added para -toluenesulfonic acid monohydrate acid (63 mg, 0.333 mmol, 3 eq.). The reaction mixture was stirred at room temperature for 24 hours. The mixture was diluted with H ₂ O and extracted with DCM. The organic extract was evaporated under reduced pressure. The crude product was purified by preparative HPLC chromatography (Method 1) to give Example 4 (9.0 mg, 8% yield) as a white solid.

Example 4 : ESIMS [MH] 966.5.

Exact weight: 967.61

^1H NMR (600 MHz, DMSO- d6 ) δ 0.59 (q, J=11.9 Hz, 1H) _, 0.74 (d, J=6.7 Hz, 3H), 0.79 (d , J=6.7 Hz, 3H), 0.80-0.90 (m, 8H), 0.92-1.00 (m, 4H), 1.04-1.10 (m, 1H), 1.12-1.34 (m, 7H), 1.34-1.67 (m, 14H), 1.69 (s, 3H), 1.71-1.78 (m, 2H), 1. 88-1.94 (m, 1H), 1.96-2.08 (m, 3H), 2.10-2.25 (m, 2H), 2.60-2.69 (m, 4H), 2.80-2.94 (m, 2H), 3.07-3.20 (m, 5H), 3.21-3.26 (m, 2H), 3.33 (s, 3H), 3. 41-3.48 (m, 1H), 3.56-3.66 (m, 3H), 3.99 (dd, J=6.5, 3.5 Hz, 1H), 4.53-4, 63 (m, 3H), 4.95-5.00 (m, 2H), 5.07 (d, J=4.8 Hz, 1H), 5.35 (s, 1H), 5.45 (dd , J=14.9, 9.8 Hz, 1H), 5.98 (d, J=11.0 Hz, 1H), 6.15 (dd, J=14.8, 10.7 Hz, 1H) , 6.23 (dd, J=14.6, 10.7 Hz, 1H), 6.44 (dd, J=14.6, 11.0 Hz, 1H).

Example 5: C16-(2-hydroxyethyl)-2-oxoimidazolidin-1-yl)-C32-deoxyrapamycin

Example 5

* Absolute stereochemical configuration for C16 is not specified

To a solution of intermediate 1 (100 mg, 0.111 mmol, 1.0 eq) and 1-(2-hydroxyethyl)imidazolidin-2-one (289 mg, 1.66 mmol, 15 eq) in DCM (2 ml) 4-methylbenzenesulfonic acid monohydrate (119 mg, 0.626 mmol, 3 eq.) was added. The reaction mixture was stirred at room temperature overnight. The reaction mixture was diluted with H ₂ O and extracted with DCM. The organic extract was evaporated under reduced pressure. The crude product was purified by preparative HPLC (method 1) followed by chiral preparative HPLC (method 2) to give Example 5 (13.7 mg, 12% yield) as a white solid.

Example 5 : ESIMS [M+H] 998.5.

Exact weight: 997.62

^1H NMR (400 MHz, DMSO- d6 ) δ 6.45 (dd, J=14.3, 11.0 Hz _, 1H), 6.23 (dd, J=14.3, 10.6 Hz, 1H), 6.14 (dd, J=14.6, 10.6 Hz, 1H), 5.98 (d, J=10.9 Hz, 1H), 5.44 (dd, J=14.5 , 9.8 Hz, 1H), 5.28 (s, 1H), 5.07 (d, J=4.7 Hz, 1H), 5.02-4.96 (m, 2H), 4.67 (t, J=5.3 Hz, 1H), 4.64-4.53 (m, 3H), 4.03-3.96 (m, 1H), 3.71-3.57 (m, 3H ), 3.52-3.42 (m, 2H), 3.42-3.29 (m, 6H), 3.22-3.07 (m, 7H), 2.95-2.79 (m , 2H), 2.71-2.56 (m, 1H), 2.27-1.87 (m, 6H), 1.80-1.36 (m, 20H), 1.36-1.12 (m, 6H), 1.11-1.02 (m, 1H), 1.00-0.91 (m, 4H), 0.91-0.76 (m, 11H), 0.74 (d , J=6.7 Hz, 3H), 0.59 (q, J=11.9 Hz, 1H).

Example 6: C16-(1,1-dioxido-1,2-thiazetidin-2-yl)-C32-deoxyrapamycin

(diastereomer 1)

Example 7: C16-(1,1-dioxido-1,2-thiazetidin-2-yl)-C32-deoxyrapamycin

(diastereomer 2)

Example 6 and Example 7

* Absolute stereochemical configuration for C16 is not specified

Diastereomer 1 . Zinc chloride was added to a solution of intermediate 1 (150 mg, 0.167 mmol, 1.0 eq) and 1,2-thiazetidine-1,1-dioxide (89 mg, 0.833 mmol, 5 eq) in DCM (3 mL). (II) (1 M solution in diethyl ether, 0.5 ml, 0.500 mmol, 3 eq.) at 0°C. The reaction mixture was stirred at room temperature for 2 hours. The mixture was diluted with H ₂ O and extracted with ethyl acetate. The organic extract was evaporated under reduced pressure. The crude product, a diastereomeric mixture, was separated by flash chromatography (silica; MeCN/DCM 0:100 to 100:0).

Final purification of the first eluted diastereomer by preparative HPLC (method 2) provided Example 6 (18 mg, 10.7% yield) as a white solid.

Example 6 : ESIMS [M+NH ₄ ] 992.6, [M+FA-H] 1019.6.

Exact weight: 974.55

^1H NMR (400 MHz, Chloroform- d ) δ 6.39 (dd, J=14.8, 9.8 Hz, 1H), 6.34 (dd, J=14.8, 10.3 Hz, 1H ), 6.18-6.10 (m, 1H), 6.02 (d, J=10.3, 1.5 Hz, 1H), 5.66 (dd, J=15.1, 8.6 Hz, 1H), 5.38 (s, 1H), 5.32 (dd, J=6.4, 1.8 Hz, 1H), 5.23 (d, J=9.6 Hz, 1H), 4.87-4.76 (m, 1H), 4.12-4.06 (m, 1H), 3.99 (ddd, J=12.0, 8.2, 6.2 Hz, 1H), 3.92-3.86 (m, 1H), 3.84 (dd, J=10.7, 4.4 Hz, 1H), 3.81-3.74 (m, 1H), 3.63- 3.59 (m, 2H), 3.59 (d, J=3.6 Hz, 1H), 3.46 (d, J=7.6 Hz, 1H), 3.43 (s, 3H), 3.40-3.37 (m, 1H), 3.32 (s, 3H), 3.04 (ddd, J=8.2, 5.8, 3.8 Hz, 1H), 3.00- 2.94 (m, 1H), 2.94-2.89 (m, 1H), 2.88-2.84 (m, 1H), 2.64 (d, J=6.7 Hz, 1H) , 2.47-2.38 (m, 1H), 2.36-2.27 (m, 2H), 2.21-2.12 (m, 1H), 2.03-1.97 (m, 1H), 1.95-1.77 (m, 7H), 1.76-1.67 (m, 5H), 1.65 (s, 3H), 1.60-1.50 (m, 6H) , 1.47-1.39 (m, 2H), 1.38-1.27 (m, 4H), 1.25-1.10 (m, 3H), 1.08-1.04 (m, 1H), 1.03-1.01 (m, 3H), 1.00 (d, J=1.4 Hz, 3H), 0.99-0.97 (m, 3H), 0.97 (d , J=6.6 Hz, 3H), 0.93 (d, J=6.8 Hz, 3H), 0.75-0.66 (m, 1H).

Final purification of the second eluted diastereomer using SFC chromatography (method 2) provided Example 7 (15.8 mg, 9.2% yield) as a white solid.

Example 7 : ESIMS [M+NH ₄ ] 992.7, [M+FA-H] 1019.6.

Exact weight: 974.55

¹ H NMR (400 MHz, Chloroform- d ) δ 6.46 (dd, J=14.2, 10.9 Hz, 1H), 6.22 (dd, J=14.2, 10.6 Hz, 1H ), 6.14 (dd, J=14.5, 10.6 Hz, 1H), 6.00 (d, J=10.9 Hz, 1H), 5.38 (dd, J=14.5, 9.6 Hz, 1H), 5.27-5.19 (m, 1H), 5.12-5.05 (m, 1H), 4.66-4.59 (m, 1H), 4.59 -4.53 (m, 1H), 4.25 (d, J=1.8 Hz, 1H), 4.17-4.06 (m, 3H), 4.02 (ddd, J=11.8 , 8.0, 3.5 Hz, 1H), 3.66 (d, J=7.0 Hz, 1H), 3.60-3.52 (m, 2H), 3.43-3.34 ( m, 4H), 3.28 (s, 3H), 3.17-3.07 (m, 1H), 3.03-2.95 (m, 1H), 2.95-2.89 (m, 1H), 2.84-2.73 (m, 1H), 2.65-2.62 (m, 1H), 2.35-2.26 (m, 1H), 2.27-2.21 ( m, 1H), 2.21-2.12 (m, 3H), 2.12-2.05 (m, 1H), 2.02-1.98 (m, 1H), 1.97-1, 95 (m, 3H), 1.91-1.83 (m, 1H), 1.82-1.70 (m, 3H), 1.70-1.51 (m, 10H), 1.50- 1.17 (m, 10H), 1.11-1.06 (m, 1H), 1.06-1.02 (m, 6H), 1.01-0.96 (m, 1H), 0. 93 (d, J=6.8 Hz, 4H), 0.89 (d, J=6.6 Hz, 3H), 0.86 (d, J=6.8 Hz, 3H), 0.64 ( q, J=11.9 Hz, 1H).

Example 8: C16-(1,1-dioxido-1,2-thiazetidin-2-yl)-C32-deoxo-C40-(2-hydroxyethoxy)rapamycin (diastereomer 1)

Example 9: C16-(1,1-dioxido-1,2-thiazetidin-2-yl)-C32-deoxo-C40-(2-hydroxyethoxy)rapamycin (diastereomer 2)

Example 8 and Example 9

* Absolute stereochemical configuration for C16 is not specified

To a solution of intermediate 3 (50 mg, 0.053 mmol, 1.0 eq) and 1,2-thiazetidine-1,1-dioxide (56.7 mg, 0.530 mmol, 10 eq) in DCM (1 ml) was added zinc(II) chloride (1 M solution in diethyl ether, 0.265 ml, 0.265 mmol, 5 eq.). The reaction mixture was stirred at room temperature for 1.5 hours. The reaction mixture was diluted with H ₂ O and extracted with ethyl acetate. The organic extract was evaporated under reduced pressure. The crude product, a diastereomeric mixture, was separated by flash chromatography (silica; MeCN/DCM 0:100 to 100:0).

Final purification of the first eluted diastereomer using preparative HPLC (method 2) provided Example 8 (3.1 mg, 5.5% yield) as a white solid.

Example 8 : ESIMS [M+NH ₄ ] 1037.0, [M+FA-H] 1064.1.

Exact weight: 1018.58

^1H NMR (400 MHz, DMSO- d6 ) δ: 6.53-6.44 (m, 1H), 6.41 ( _dd , J=11.0, 5.6 Hz, 1H), 6.29 -6.14 (m, 2H), 6.07 (dd, J=11.0, 1.5 Hz, 1H), 5.54 (dd, J=14.6, 9.1 Hz, 1H), 5.06 (d, J=8.9 Hz, 1H), 4.99-4.93 (m, 1H), 4.92-4.83 (m, 1H), 4.70-4.60 ( m, 1H), 4.49-4.42 (m, 1H), 4.13-4.02 (m, 3H), 3.92-3.86 (m, 1H), 3.69-3, 64 (m, 1H), 3.59-3.54 (m, 1H), 3.52-3.42 (m, 5H), 3.33 (s, 3H), 3.30-3.27 ( m, 1H), 3.13 (s, 3H), 3.10-3.02 (m, 2H), 3.02-2.94 (m, 2H), 2.90-2.80 (m, 1H), 2.30-2.21 (m, 1H), 2.21-2.10 (m, 1H), 2.10-2.00 (m, 2H), 2.00-1.87 ( m, 3H), 1.82 (s, 3H), 1.81-1.62 (m, 6H), 1.58-1.54 (m, 1H), 1.53-1.47 (m, 5H), 1.47-1.35 (m, 5H), 1.36-1.23 (m, 2H), 1.23-1.09 (m, 4H), 1.06-1.01 ( m, 1H), 1.01-0.95 (m, 6H), 0.90-0.87 (m, 1H), 0.87-0.83 (m, 3H), 0.81 (d, J=6.7 Hz, 3H), 0.76-0.70 (m, 4H), 0.68-0.60 (m, 1H).

Final purification of the second eluted diastereomer using preparative HPLC (method 2) provided Example 9 (2.8 mg, 4.9% yield) as a white solid.

Example 9: ESIMS [M+NH4] 1037.2, [M+FA-H] 1064.3.

Exact weight: 1018.58

^1H NMR (400 MHz, DMSO- d6 ) δ: 6.48-6.40 (m, 1H) _, 6.27-6.19 (m, 1H), 6.19-6.13 (m, 1H), 6.10-6.05 (m, 1H), 5.52 (dd, J=14.2, 9.3 Hz, 1H), 5.13-5.00 (m, 2H), 4 .96 (d, J=9.4 Hz, 1H), 4.67-4.57 (m, 1H), 4.48-4.40 (m, 1H), 4.17-4.10 (m , 2H), 4.10-4.02 (m, 2H), 3.98-3.90 (m, 1H), 3.56-3.49 (m, 2H), 3.50-3.43 (m, 4H), 3.37 (d, J=11.2 Hz, 1H), 3.33 (s, 3H), 3.23-3.15 (m, 1H), 3.15-3, 10 (m, 4H), 3.07-2.95 (m, 2H), 2.75-2.64 (m, 1H), 2.27-2.14 (m, 2H), 2.11- 2.00 (m, 2H), 2.00-1.89 (m, 3H), 1.82 (s, 3H), 1.76-1.65 (m, 2H), 1.65-1, 56 (m, 3H), 1.56-1.49 (m, 7H), 1.48-1.36 (m, 3H), 1.34-1.23 (m, 3H), 1.23- 1.04 (m, 5H), 1.01-0.94 (m, 4H), 0.90 (d, J=6.4 Hz, 3H), 0.88-0.83 (m, 5H) , 0.80 (d, J=6.7 Hz, 3H), 0.75 (d, J=6.7 Hz, 3H), 0.62 (q, J=11.8 Hz, 1H).

Example 10: C16-(1,1-dioxido-1,2-thiazetidin-2-yl)-C32-deoxo-C40-dimethylphosphinylrapamycin (diastereomer 1)

Example 11: C16-(1,1-dioxido-1,2-thiazetidin-2-yl)-C32-deoxo-C40-dimethylphosphinylrapamycin (diastereomer 2)

Example 10 and Example 11

* Absolute stereochemical configuration for C16 is not specified

To a solution of intermediate 4 (100 mg, 0.102 mmol, 1.0 eq) and 1,2-thiazetidine-1,1-dioxide (54.9 mg, 0.512 mmol, 5 eq) in DCM (5 ml) was added zinc(II) chloride (1 M solution in diethyl ether, 0.512 ml, 0.512 mmol, 5 eq.). The reaction mixture was stirred at room temperature for 15 minutes. The reaction mixture was diluted with H ₂ O and extracted with ethyl acetate. The organic extract was evaporated under reduced pressure. The crude product, a diastereomeric mixture, was separated using SFC chromatography (method 3).

Final purification of the first eluted diastereomer using preparative HPLC (Method 2) provided Example 10 (20.7 mg, 18.8% yield) as a white solid.

Example 10 : ESIMS [M+H] 1051.9, [M+FA-H] 1096.0.

Exact weight: 1050.56

^1H NMR (400 MHz, DMSO- d6 ) δ 6.49-6.39 (m, _2H ), 6.30-6.14 (m, 2H), 6.11-6.01 (m, 1H ), 5.60-5.50 (m, 1H), 5.06-5.00 (m, 1H), 4.98-4.93 (m, 1H), 4.87 (d, J=9 .7 Hz, 1H), 4.67-4.58 (m, 1H), 4.17-4.01 (m, 3H), 4.01-3.92 (m, 1H), 3.92- 3.85 (m, 1H), 3.73-3.64 (m, 1H), 3.60-3.52 (m, 1H), 3.50-3.42 (m, 1H), 3. 35-3.26 (m, 4H), 3.15-3.12 (m, 3H), 3.11-3.01 (m, 2H), 3.01-2.95 (m, 1H), 2.88-2.79 (m, 1H), 2.29-2.20 (m, 1H), 2.20-2.12 (m, 1H), 2.10-1.99 (m, 2H ), 1.99-1.87 (m, 3H), 1.82 (s, 3H), 1.80-1.71 (m, 1H), 1.71-1.62 (m, 4H), 1.58-1.54 (m, 1H), 1.52-1.48 (m, 5H), 1.47-1.31 (m, 13H), 1.28-1.11 (m, 4H ), 1.07-1.02 (m, 1H), 1.01-0.95 (m, 8H), 0.87-0.84 (m, 3H), 0.82 (d, J=6 .7 Hz, 3H), 0.79-0.76 (m, 1H), 0.74 (d, J=6.7 Hz, 3H), 0.72-0.66 (m, 1H).

Final purification of the second eluted diastereomer using preparative HPLC (Method 2) provided Example 11 (13.4 mg, 12.2% yield) as a white solid.

Example 11 : ESIMS [M+H] 1051.8, [M+FA-H] 1095.8.

Exact weight: 1050.56

¹ H NMR (400 MHz, DMSO- d ₆ ) δ) 6.51-6.39 (m, 1H), 6.28-6.13 (m, 2H), 6.08 (d, J=11, 1 Hz, 1H), 6.04-5.88 (m, 1H), 5.57-5.46 (m, 1H), 5.05 (s, 1H), 5.03 (d, J=5 .7 Hz, 1H), 4.96 (d, J=9.9 Hz, 1H), 4.67-4.56 (m, 1H), 4.18-4.11 (m, 2H), 4 .09-4.02 (m, 2H), 3.99-3.89 (m, 2H), 3.60-3.49 (m, 2H), 3.37 (d, J=13.3 Hz , 1H), 3.30 (s, 3H), 3.18 (dt, J=7.7, 5.1 Hz, 1H), 3.15-3.11 (m, 4H), 3.07- 3.00 (m, 1H), 2.73-2.63 (m, 1H), 2.28-2.13 (m, 2H), 2.13-2.05 (m, 2H), 2. 05-1.97 (m, 2H), 1.97-1.88 (m, 1H), 1.83 (s, 3H), 1.80-1.69 (m, 1H), 1.68- 1.58 (m, 4H), 1.58-1.48 (m, 7H), 1.48-1.43 (m, 1H), 1.42-1.34 (m, 10H), 1. 34-1.28 (m, 1H), 1.28-1.22 (m, 2H), 1.22-1.13 (m, 2H), 1.13-1.03 (m, 1H), 1.03-0.94 (m, 5H), 0.94-0.88 (m, 4H), 0.86 (d, J=6.5 Hz, 3H), 0.81 (d, J= 6.7 Hz, 3H), 0.75 (d, J=6.6 Hz, 3H), 0.66 (q, J=12.0 Hz, 1H).

Example 12: C16-(1,1-dioxido-1,2-thiazetidin-2-yl)-C32-deoxo-C40-( S )-(1H-tetrazol-1-yl)rapamycin (diastereomer 1)

Example 13: C16-(1,1-dioxido-1,2-thiazetidin-2-yl)-C32-deoxo-C40-( S )-(1H-tetrazol-1-yl)rapamycin (diastereomer 2)

Example 12 and Example 13

* Absolute stereochemical configuration for C16 is not specified

To a solution of intermediate 5 (118 mg, 0.124 mmol, 1.0 eq.) and 1,2-thiazetidine 1,1-dioxide (66.4 mg, 0.620 mmol, 5 eq.) in DCM (6 mL) was added zinc(II) (1 M solution in diethyl ether, 0.620 ml, 0.620 mmol, 5 eq.). The reaction mixture was stirred at room temperature for three hours. The reaction mixture was diluted with H ₂ O and extracted with ethyl acetate. The organic extract was evaporated under reduced pressure. The crude product, a diastereomeric mixture, was separated using SFC chromatography (method 3).

Final purification of the first eluted diastereomer using preparative HPLC (method 2) provided Example 12 (14.8 mg, 11.3% yield) as a white solid.

Example 12: ESIMS [M+NH4] 1050.0, [M+FA-H] 1072.1.

Exact weight: 1026.57

^1H NMR (400 MHz, DMSO- d6 ) δ 9.32 (s, 1H), 6.52 (s, 1H), _6.45-6.37 (m, 1H), 6.31-6, 14 (m, 2H), 6.10-6.04 (m, 1H), 5.60-5.49 (m, 1H), 5.23-5.12 (m, 1H), 5.07- 5.01 (m, 1H), 4.99-4.93 (m, 1H), 4.87 (d, J=9.7 Hz, 1H), 4.69-4.59 (m, 1H) , 4.15-4.02 (m, 3H), 3.94-3.84 (m, 1H), 3.72-3.54 (m, 3H), 3.51-3.42 (m, 1H), 3.27 (s, 4H), 3.11 (s, 3H), 3.07 (dd, J=7.3, 5.1 Hz, 1H), 3.01-2.91 (m , 1H), 2.85 (td, J=10.6, 9.5, 4.7 Hz, 1H), 2.28-2.13 (m, 3H), 2.10-2.00 (m , 2H), 1.98-1.88 (m, 3H), 1.82 (s, 3H), 1.80-1.75 (m, 1H), 1.73-1.67 (m, 4H ), 1.59-1.53 (m, 4H), 1.53-1.45 (m, 6H), 1.45-1.34 (m, 3H), 1.33-1.20 (m , 3H), 1.20-1.07 (m, 3H), 1.05-1.01 (m, 1H), 1.00-0.94 (m, 6H), 0.88-0.82 (m, 6H), 0.80-0.75 (m, 1H), 0.73 (d, J=6.7 Hz, 3H).

Final purification of the second eluted diastereomer using preparative HPLC (Method 2) provided Example 13 (12.3 mg, 9.4% yield) as a white solid.

Example 13 : ESIMS [M+H] 1027.7, [MH] 1025.6, [M+FA-H] 1071.6.

Exact weight: 1026.57

^1H NMR (400 MHz, DMSO- d ₆ δ 9.30 (s, 1H), 6.52-6.38 (m, 1H), 6.28-6.14 (m, 2H), 6.08 (d, J=11.1 Hz, 1H), 5.58-5.42 (m, 1H), 5.21-5.14 (m, 1H), 5.10-5.00 (m, 2H ), 5.00-4.92 (m, 1H), 4.69-4.57 (m, 1H), 4.16-4.10 (m, 3H), 4.08-4.01 (m , 1H), 3.97-3.91 (m, 1H), 3.61 (dt, J=10.6, 4.1 Hz, 1H), 3.57-3.51 (m, 1H), 3.51-3.46 (m, 1H), 3.46-3.36 (m, 1H), 3.27 (s, 3H), 3.19-3.15 (m, 1H), 3. 12 (s, 3H), 3.11-3.08 (m, 1H), 2.80-2.68 (m, 1H), 2.25-2.15 (m, 3H), 2.10- 2.01 (m, 2H), 2.01-1.89 (m, 2H), 1.83 (s, 3H), 1.79-1.65 (m, 4H), 1.64-1, 58 (m, 2H), 1.57-1.54 (m, 2H), 1.54-1.47 (m, 7H), 1.46-1.35 (m, 3H), 1.32- 1.18 (m, 5H), 1.14-1.04 (m, 3H), 0.98 (d, J=6.6 Hz, 3H), 0.91-0.85 (m, 7H) , 0.83 (d, J=6.7 Hz, 3H), 0.72 (d, J=6.6 Hz, 3H).

Example 14: C16-(1,1-dioxidoizothiazolidin-2-yl)-C32-deoxo-C40-dimethylphosphinylrapamycin (diastereomer 1)

Example 15: C16-(1,1-dioxidoisothiazolidin-2-yl)-C32-deoxo-C40-dimethylphosphinylrapamycin (diastereomer 2)

Example 14 and Example 15

* Absolute stereochemical configuration for C16 is not specified

Dissolved intermediate 4 (0.146 g, 0.150 mmol) and isothiazolidine-1,1-dioxide (0.181 g, 1.496 mmol) in anhydrous acetonitrile (1.5 ml). Para -toluenesulfonic acid monohydrate (2.84 mg, 0.015 mmol) was added in one portion. The reaction mixture was stirred at room temperature for two hours.

A saturated aqueous solution of NaHCO ₃ was added. The mixture was extracted several times with ethyl acetate. The organic extracts were combined, dried over Na ₂ SO, decanted and concentrated to give the crude product as a colorless gum.

The crude product was dissolved in MeOH (2.5 ml) and purified in one run using reverse phase preparative chromatography (40 to 90% acetonitrile in water plus 0.1% TFA modifier on a C18 ISCO 100 g column).

Fractions corresponding to the first elution peak were pooled and reduced to approximately 1/3 volume on a rotary evaporator. The remaining solution was rebasified with saturated aqueous NaHCO ₄ and extracted several times with EtOAc. The organic extracts were combined, dried over Na ₂ SO ₄ , decanted and concentrated to give Example 14 (0.055 g, 0.044 mmol, 29.3% yield) as a white solid.

Example 14 : ESIMS [M+NH4] 1082.8, [MH] 1063.7.

HRMS: calculated for C55H89N2O14PSNa as an adduct with sodium - 1087.5670. Discovered - 1087.5725.

^1H NMR (400 MHz, Chloroform- d ) δ 6.45 (dd, J =14.4, 10.9 Hz, 1H), 6.22 (dd, J =14.5, 10.6 Hz, 1H ), 6.13 (dd, J =14.6, 10.5 Hz, 1H), 5.98 (d, J =10.8 Hz, 1H), 5.35 (dd, J =14.7, 9.8 Hz, 1H), 5.23-5.17 (m, 1H), 5.10 (d, J =9.8 Hz, 1H), 4.67 (m, 2H), 4.47 ( d, J =1.8 Hz, 1H), 4.18-4.04 (m, 2H), 4.03-3.91 (m, 1H), 3.72 (d, J =6.5 Hz , 1H), 3.68-3.48 (m, 2H), 3.38 (m, 4H), 3.28 (s, 3H), 3.26-3.12 (m, 2H), 3, 12-2.91 (m, 4H), 2.74 (m, 1H), 2.48-2.26 (m, 3H), 2.26-2.12 (m, 3H), 2.12- 2.04 (m, 2H), 1.87 (s, 3H), 1.85-1.72 (m, 4H), 1.72-1.54 (m, 12H), 1.54-1, 43 (m, 6H), 1.43-1.33 (m, 3H), 1.33-1.21 (m, 2H), 1.21-1.09 (m, 2H), 1.03 ( m, 7H), 1.00-0.78 (m, 9H), 0.72 (q, J =11.9 Hz, 1H).

Fractions corresponding to the second elution peak were pooled and reduced to approximately 1/3 volume on a rotary evaporator. The basicity of the remaining solution was increased using a saturated aqueous solution of NaHCO3. The mixture was extracted several times with ethyl acetate. The organic extracts were combined, dried over Na2SO4, decanted and concentrated to give Example 15 (0.010 g, 7.51 µmol, 5.02% yield) as a white solid.

Example 15 : ESIMS [M+NH4] 1082.8, [MH] 1063.8.

^1H NMR (400 MHz, Chloroform- d ) δ 6.36 (dd, J =19.2, 10.3 Hz, 1H), 6.13 (m, 1H), 6.04-5.84 (m , 1H), 5.65 (m, 1H), 5.32 (m, 1H), 6.21 (m, 1H), 5.11 (m, 1H), 4.81 (dd, J =13, 7, 7.5 Hz, 1H), 4.21-4.03 (m, 1H), 3.83 (dd, J =15.2, 5.1 Hz, 2H), 3.74 (d, J =13.2 Hz, 1H), 3.59 (dq, J =10.9, 6.8, 5.6 Hz, 2H), 3.52 (d, J =7.1 Hz, 1H), 3 .46-3.33 (m, 7H), 3.30 (m, 3H), 3.18 (m, 3H), 3.10-2.82 (m, 3H), 2.39 (t, J =4.1 Hz, 1H), 2.28 (m, 3H), 2.14 (m, 3H), 1.90-1.77 (m, 4H), 1.74 (m, 4H), 1 .66 (d, J =10.4 Hz, 3H), 1.61-1.41 (m, 12H), 1.34-1.19 (m, 5H), 1.18-1.11 (m , 1H), 1.11-0.79 (m, 19H), 0.78-0.68 (m, 1H).

Example 16 : (R)-C16-(1,1-dioxidoisothiazolidin-2-yl)-C32-deoxo-C40-(2-hydroxyethoxy)rapamycin

Example 16

The absolute stereochemical configuration at C16 is known to be ( R ) based on the starting material used in Example 2 .

Example 16 was prepared by a two-step procedure from 2-((tert-butyldimethylsilyl)oxy)ethanol and Example 2 .

Step 1: Preparation of Intermediate C

Dissolve 2-((tert-butyldimethylsilyl)oxy)ethanol (0.142 g, 0.803 mmol) in anhydrous toluene (0.4 ml). N,N -diisopropylethylamine (0.147 mL, 0.843 mmol) was added via syringe. The mixture was evacuated with nitrogen, then cooled to 0°C in an ice bath. Triflate anhydride (0.132 mL, 0.779 mmol) was added via syringe over a period of 60 seconds. The reaction mixture was stirred at 0°C for 30 minutes.

N,N -Diisopropylethylamine (0.147 ml, 0.843 mmol), toluene (0.5 ml) and Example 2 (0.198 g, 0.200 mmol) were added. The ice bath was removed and the reaction mixture was stirred overnight at 40°C and then for one hour at 55°C.

The reaction mixture was diluted with saturated aqueous NaHCO3 and extracted four times with EtOAc. The organic extracts were combined, dried over Na2SO4, vacuum filtered through celite and concentrated to give the crude product as a colorless oil.

The crude product was purified by silica gel flash column chromatography (0 to 40% acetone in heptane elution gradient, 12 g silica column, TLC in 30% acetone in heptane, UV visualized) to give intermediate C (0.077 g, 0.067 mmol, 33.5% yield) as a white solid.

Intermediate C : ESIMS [M+NH4] 1164.7, [MH] 1147.0.

Stage 2. Obtaining example 16

Combine intermediate C (0.077 g, 0.067 mmol) with pyridine (5.43 μL, 0.067 mmol) in anhydrous THF (0.7 mL). The mixture was evacuated twice with nitrogen and then cooled to 0°C in an ice bath. HF-pyridine (0.086 mL, 0.671 mmol) was added dropwise using a syringe over a period of 15 seconds. The reaction mixture was stirred at 0°C for 70 minutes.

The reaction mixture was quenched with saturated aqueous NaHCO3 and extracted several times with EtOAc. The organic extracts were combined, dried over Na2SO4, vacuum filtered through celite and concentrated to give a white solid crude product.

The crude product was purified by silica gel flash column chromatography (0 to 40% acetone in heptane elution gradient, 12 g silica column, TLC in 40% EtOAc in heptane, visualized by UV) to give Example 16 (0.049 g , 0.046 mmol, yield 69.0%) as a white solid.

Example 16 : ESIMS [M+NH4] 1050.9, [MH] 1031.9.

¹ H NMR (600 MHz, Chloroform- d ) δ 6.45 (dd, J =14.6, 10.9 Hz, 1H), 6.22 (dd, J =14.7, 10.6 Hz, 1H ), 6.13 (dd, J =14.9, 10.6 Hz, 1H), 5.98 (d, J =10.9 Hz, 1H), 5.35 (dd, J =14.9, 9.8 Hz, 1H), 5.20 (d, J =5.9 Hz, 1H), 5.09 (d, J =9.9 Hz, 1H), 4.71 (d, J =12, 3 Hz, 1H), 4.62 (m, 1H), 4.46 (s, 1H), 4.11 (d, J =6.8 Hz, 1H), 3.96 (t, J =11, 3 Hz, 1H), 3.78 (m, 1H), 3.70 (m, 3H), 3.63-3.55 (m, 3H), 3.43 (s, 3H), 3.28 ( s, 3H), 3.24-3.15 (m, 2H), 3.12-3.04 (m, 2H), 3.07-2.98 (m, 1H), 2.97 (q, J =7.9 Hz, 1H), 2.75 (m, 1H), 2.44-2.33 (m, 1H), 2.30 (M, 2H), 2.23 (m, 1H), 2.16 (m, 2H), 2.09 (d, J =13.1 Hz, 1H), 2.01 (m, 1H), 1.88 (s, 3H), 1.84 (d, J =13.1 Hz, 1H), 1.78-1.70 (m, 3H), 1.70-1.62 (m, 4H), 1.52 (dd, J =11.8, 7.7 Hz, 1H), 1.50-1.41 (m, 2H), 1.43-1.32 (m, 1H), 1.34-1.27 (m, 1H), 1.30- 1.17 (m, 8H), 1.04 (dd, J =9.5, 6.6 Hz, 7H), 1.02-0.85 (m, 10H), 0.85 (d, J = 6.8 Hz, 3H), 0.69 (q, J =12.0 Hz, 1H).

Example 17 : C16-(1,1-dioxidoizothiazolidin-2-yl)-C32-deoxo-C40-( S )-(1H-tetrazol-1-yl)rapamycin

Example 17

* Absolute stereochemical configuration for C16 is not specified

Combine intermediate 5 (0.109 g, 0.114 mmol) with isothiazolidine-1,1-dioxide (0.139 g, 1.145 mmol) in anhydrous acetonitrile (1.1 ml). Para -toluenesulfonic acid monohydrate (0.0022 g, 0.011 mmol) was added. The reaction mixture was stirred at room temperature for two hours.

The reaction mixture was diluted with saturated aqueous NaHCO3 solution. The mixture was extracted several times with EtOAc. The organic extracts were combined, dried over Na2SO, decanted and concentrated to give the crude product as a yellow gum.

The crude product was purified by silica gel flash column chromatography (0 to 40% acetone/heptane elution gradient, 24 g silica column, TLC in 40% acetone/heptane, UV visualized) to give Example 17 (0.053 g , 0.046 mmol, yield 40.0%) as a white solid.

Example 17 : ESIMS [M+H] 1041.8, [MH] 1039.8.

HRMS: calculated for C54H84N6O12SNa - 1063.5765. Discovered - 1063.5759.

^1H NMR (600 MHz, Chloroform- d ) δ 8.86 (s, 1H), 6.39 (dd, J =14.7, 11.0 Hz, 1H), 6.22 (dd, J =14 .7, 10.7 Hz, 1H), 6.11 (dd, J =15.0, 10.5 Hz, 1H), 5.99 (d, J =10.9 Hz, 1H), 5.34 (dd, J =14.9, 9.8 Hz, 1H), 5.18 (d, J =5.7 Hz, 1H), 5.10 (d, J =9.8 Hz, 1H), 4 .87 (m, 1H), 4.67 (m, 1H), 4.63 (d, J =12.2 Hz, 1H), 4.50 (s, 1H), 4.13 (d, J = 6.1 Hz, 1H), 3.97 (t, J =11.4 Hz, 1H), 3.78 (d, J =6.2 Hz, 1H), 3.69-3.62 (m, 1H), 3.55 (dt, J =11.2, 4.0 Hz, 1H), 3.50-3.39 (m, 1H), 3.39 (s, 3H), 3.38-3 .30 (m, 1H), 3.27 (s, 3H), 3.25-3.13 (m, 1H), 3.07 (td, J =8.4, 3.8 Hz, 1H), 3.01 (m, 1H), 2.96 (q, J =7.9 Hz, 1H), 2.70-2.63 (m, 2H), 2.39 (m, 1H), 2.35 -2.21 (m, 3H), 2.18 (d, J =13.7 Hz, 1H), 2.16-2.09 (m, 1H), 1.95-1.81 (m, 3H ), 1.82 (s, 3H), 1.75 (m, 3H), 1.65 (s, 3H), 1.60 (m, 6H), 1.58-1.50 (m, 1H) , 1.53-1.34 (m, 2H), 1.32 (m, 1H), 1.29 (m, 1H), 1.25 (m, 6H), 1.12 (m, 1H), 1.05 (d, J =6.6 Hz, 3H), 1.05-0.78 (m, 12H).

Example 18: C16-(1,1-dioxidoisothiazolidin-2-yl)-C32-deoxo-C40-(2-ethoxyethoxy)rapamycin

Example 18

* Absolute stereochemical configuration for C16 is not specified

Combine intermediate 6 (0.090 g, 0.093 mmol) with isothiazolidine-1,1-dioxide (0.112 g, 0.926 mmol) in anhydrous dichloromethane (0.93 ml). Para -toluenesulfonic acid monohydrate (1.761 mg, 9.26 µmol) was added in one portion. The reaction mixture was stirred at room temperature for 100 minutes.

The entire reaction mixture was purified by silica gel flash column chromatography (0 to 40% acetone in heptane elution gradient, 24 g silica column, TLC in 40% acetone in heptane, visualized by UV) to give the mixture of Example 18 and residual unreacted isothiazolidine-1,1-dioxide.

The stirred material was again purified by silica gel flash column chromatography (0 to 50% acetonitrile in dichloromethane elution gradient, 24 g column, TLC in 30% acetonitrile in dichloromethane, visualized by UV) to give Example 18 (0.021 g, 0.019 mmol, yield 20.4%) as a white solid.

Example 18 : ESIMS [M+NH4] 1078.9, [MH] 1059.9.

HRMS: calculated for C57H92N2O14SNa, adduct with sodium - 1083.6167. Discovered - 1083.6151.

^1H NMR (600 MHz, Chloroform- d ) δ 6.45 (dd, J =14.5, 11.0 Hz, 1H), 6.21 (dd, J =14.6, 10.7 Hz, 1H ), 6.14 (dd, J =14.8, 10.8 Hz, 1H), 5.98 (d, J =10.9 Hz, 1H), 5.35 (dd, J =14.8, 9.8 Hz, 1H), 5.20 (d, J =5.8 Hz, 1H), 5.09 (d, J =9.8 Hz, 1H), 4.72 (d, J =12, 1 Hz, 1H), 4.61 (m, 1H), 4.46 (s, 1H), 4.10 (d, J =6.9 Hz, 1H), 3.96 (t, J =11, 4 Hz, 1H), **3.73 (t, J =5.5 Hz, 2H), 3.68 (d, J =6.9 Hz, 1H), 3.59 (m, 4H), 3 .53 (m, 2H), 3.46 (s, 3H), 3.28 (s, 3H), 3.21 (m, 1H), 3.13 (m, 1H), 3.07 (m, 2H), 3.05-2.99 (m, 1H), 2.97 (q, J =8.0 Hz, 1H), 2.78 (dq, J =14.4, 7.1 Hz, 1H ), 2.44-2.33 (m, 1H), 2.31 (m, 2H), 2.27-2.19 (m, 1H), 2.19-2.12 (m, 2H), 2.07-1.94 (m, 2H), 1.88 (s, 3H), 1.83 (d, J =12.9 Hz, 1H), 1.74 (m, 3H), 1.66 (m, 7H), 1.56-1.44 (m, 1H), 1.43 (m, 2H), 1.41-1.15 (m, 12H), 1.04 (m, 7H), 0.91 (m, 8H), 0.84 (d, J =6.7 Hz, 3H), 0.70 (q, J =12.0 Hz, 1H).

Example 19 : C16-(1,1-dioxidoizothiazolidin-2-yl)-C32-deoxo-C40-((3-hydroxy-2-(hydroxymethyl)-2-methylpropanoyl)oxy)rapamycin

Example 19

* Absolute stereochemical configuration for C16 is not specified

Combine intermediate 7 (0.088 g, 0.087 mmol) with isothiazolidine-1,1-dioxide (0.105 g, 0.866 mmol) in anhydrous dichloromethane (0.87 ml). Para -toluenesulfonic acid monohydrate (1.647 mg, 8.66 µmol) was added. The reaction mixture was stirred at room temperature for 30 minutes.

The entire reaction mixture was purified by normal phase flash column chromatography (0 to 60% acetonitrile in dichloromethane elution gradient, 12 g silica column, TLC in 30% acetonitrile in dichloromethane) to give Example 19 (0.020 g, 0.017 mmol , yield 19.9%) as a white solid.

Example 19 : ESIMS [M+NH4] 1123.0, [MH] 1104.0.

HRMS: Calculated for ammonium adduct, C58H92N2O16SNH4-1122.6511; discovered - 1122.652.

^1H NMR (600 MHz, Chloroform- d ) δ 6.45 (dd, J =14.7, 10.9 Hz, 1H), 6.22 (dd, J =14.7, 10.7 Hz, 1H ), 6.13 (dd, J =14.9, 10.6 Hz, 1H), 5.99 (d, J =10.9 Hz, 1H), 5.35 (dd, J =14.9, 9.8 Hz, 1H), 5.21 (d, J =5.8 Hz, 1H), 5.10 (d, J =9.9 Hz, 1H), 4.73 (ddd, J =11, 0, 9.2, 4.7 Hz, 1H), 4.69 (d, J =12.4 Hz, 1H), 4.64 (m, 1H), 4.49 (s, 1H), 4, 13 (d, J =6.5 Hz, 1H), 3.97 (t, J =11.3 Hz, 1H), 3.84 (dd, J =18.5, 11.4 Hz, 2H), 3.78-3.69 (m, 2H), 3.64 (m, 2H), 3.56 (td, J =13.5, 3.0 Hz, 1H), 3.43-3.31 ( m, 4H), 3.28 (m, 3H), 3.26-3.17 (m, 2H), 3.12-2.92 (m, 3H), 2.76-2.68 (m, 1H), 2.45-2.34 (m, 1H), 2.31-2.21 (m, 3H), 2.20-2.10 (m, 3H), 2.06 (m, 1H) , 1.87 (m, 4H), 1.75 (m, 3H), 1.71-1.59 (m, 3H), 1.60 (s, 3H), 1.60-1.50 (m , 1H), 1.48-1.39 (m, 1H), 1.42-1.31 (m, 2H), 1.33-1.20 (m, 8H), 1.11 (s, 3H ), 1.11-1.05 (m, 2H), 1.03 (dd, J =18.2, 6.6 Hz, 6H), 0.99-0.82 (m, 10H), 0, 79 (q, J =12.0 Hz, 1H).

Example 20: Biological Assays and Data

The activity of the compound of the present invention was assessed using the following in vitro and in vivo methods.

Determination of pharmacological characteristics

Materials and methods

Cell-based assay for determining rapalogue activity . Rapalogue activity was determined using an assay based on TSC1−/− MEF cells. TSC1-/- MEF cells are mouse embryonic fibroblasts deficient in the tuberous sclerosis protein TSC1, which negatively regulates mTORC1 signaling, and thus exhibit constitutive activation of mTORC1, leading to phosphorylation (activation) of downstream molecules. This cell-based assay is used to measure inhibition (dephosphorylation) of S6 and 4EBP1 by rapalogues or other mTOR inhibitors.

TSC1−/− MEF cells were seeded into poly-D-lysine-coated clear-bottom Greiner 384-well plates and incubated overnight at 37°C, 5% CO ₂ . The next day, the cells were washed 8 times with a “severe starvation” solution (1 L DPBS + 1 g D(+)-glucose + 10 ml 7.5% sodium bicarbonate solution + 20 ml 1 M HEPES) and incubated for another 2 hours in the same solution. Cells were then treated with compounds at decreasing concentrations (8 levels at 3.16-fold dilution) and incubated for 2 hours at 37°C, 5% CO ₂ . Cells were fixed with 4% paraformaldehyde for 30 min. and washed 5 times with TBS-EDTA followed by immunostaining with fluorescently tagged antibodies to pS6 (Ser240/244) (Cell Signaling #9468) and p4EBP1 (Thr 37/46) (Cell Signaling #5123). Nuclei were made visible using Hoechst stain (ThermoFisher Scientific #H3570). Cells were imaged (InCell 600) using appropriate fluorescent channels and mTOR inhibitor activity was determined using IC ₅₀ (nM) of pS6.

Animal management, compound treatment and tissue collection . All procedures involving animals were approved by the Institutional Animal Care and Use Committee of the Novartis Biomedical Research Institute, Cambridge, MA, USA. Adult male Sprague Dawley (SD) rats were purchased from Envigo (Indianapolis, USA) or Charles River (USA). After transport, rats were kept in separate, sterile, temperature- and light-controlled rooms (22°C, 12-hour light/12-hour dark cycle: lights on at 06:00 a.m./lights off at 18:00 p.m.) and with access to food and water ad libitum. Rats were acclimated for at least 3 days before experiments.

Example 2 and RAD001 were formulated for oral administration ( per os , po) doses. Blank formulations (without Example 2 or RAD001) served as vehicle controls. Rats received a single dose of Example 2, RAD001, or appropriate po vehicle. At predetermined time points after treatment, rats were anesthetized with 3.5% isoflurane and euthanized. Various organs were collected and frozen in liquid nitrogen. Blood was collected via tail vein or cardiac puncture (last collection) and frozen for further pharmacokinetic analyses. All tissues were stored at -80°C until analysis.

Determination of concentration values of example 2 and RAD001 in blood and tissues. Concentration values were determined using HPLC/mass spectrometry.

Protein extraction and immunoblotting . For protein extraction, snap-frozen tissues were lysed in MSD lysis buffer (MSD, Rockville, MD), supplemented with cOmplete protease inhibitor without EDTA and PhosSTOP phosphatase inhibitor tablets (Roche, Mannheim, Germany), and centrifuged at 13,000 g for 20 min. at 4°C. The resulting supernatant was used for immunoblotting. Protein was quantified by BCA protein assay (Thermo Scientific, MA). Samples were separated on 4-20% pre-polymerized Criterion™ TGX™ protein electrophoresis midi-gels (Bio-Rad, CA) and transferred to nitrocellulose membranes (Bio-Rad, CA) using the Trans-Blot Turbo system (Bio-Rad, CA) California). Immunoblotting was performed using antibodies to p-S6 and t-S6 from Cell Signaling Technologies (all 1:1000 in TBS-T with 5% BSA). The prefixes "p" and "t" denote the "phosphorylated" and "general" forms, respectively. HRP-conjugated secondary anti-rabbit IgG antibody (#7074) was obtained from Cell Signaling Technologies, Massachusetts. The chemiluminescence signal was generated using SuperSignal™ West Femto Enhanced Chemiluminescence Substrate (#34095, Thermo Scientific, MA) or Western Lightning® Plus-ECL Enhanced Chemiluminescence Substrate (NEL103001EA, Perkin Elmer, MA) and recorded using a ChemiDoc MP Imaging System (Bio- Rad). The resulting digital images were converted to TIFF format and subjected to quantitative analysis using ImageJ software.

Generation of FKPB12 knockdown cells . A CRISPR/CAS9 vector containing a gRNA sequence targeting the C terminus of FKBP12 (GCTTCTAAAACTGGAATGAC) was transfected into 293T cells. Selection was performed using puromycin for 48 hours and cells were plated to form colonies. FKBP12 knockdown clones were screened by immunoblotting using anti-FKBP12 antibody (Thermo Scientific, Pierce #PA1-026A). Clones with significantly reduced levels of FKBP12 were selected (approximately 80% reduction relative to levels in untreated 293T cells).

Generation of FKPB12 knockout cells . The CRISPR/Cas9 system was used to deliver ribonucleoprotein complexes containing a guide RNA (gRNA) sequence targeting FKBP12 (GCCACTACTCACCGTCTCCT) into 293T cells using the Amaxa® 4D-Nucleofector™ X kit (Lonza, V4XC-2032). Cell clones were screened by immunoblotting using an anti-FKBP12 antibody (Novus, NB300-508), and individual clones were selected that exhibited complete knockout of FKBP12 (no measurable amount of FKBP12).

Treatment of FKBP12 knockdown and FKBP12 knockout wild type (WT) 293T cells with RAD001 and Example 2. FKBP12 knockdown and FKBP12 knockout 293T WT cells were seeded at a density of 30,000 cells per well in poly-D-lysine-coated 96-wells. plates (Corning, no. 354461) in Dulbecco's modified Eagle's medium (ThermoFisher, no. 11995-065) supplemented with 10% fetal bovine serum (ThermoFisher, no. 16140-071). Cells were incubated at 37°C, 5% CO ₂ for 48 hours until they reached ~80% confluence. Cells were treated with RAD001 and Example 2 using a 12-point dose range from 1000 nM to 0.0033 nM for 2 hours at 37°C in duplicate. Medium supplemented with dimethyl sulfoxide (DMSO) blank was used as a control for both compounds. The amounts of phosphorylated S6K1 (Thr389) were determined using a sandwich ELISA kit (Cell signaling, no. 7063C) according to the manufacturer's instructions.

SPR assay to determine binding affinity to FK506-binding proteins (FKBPs) .

avi-his6 N -terminally tagged FKBPs fused to FKBP12, FKBP51, and FKBP52 were expressed in E. coli and purified by standard chromatography. Each protein was sequentially immobilized on a streptavidin chip in Biacore 8K SPR equipment (GE Healthcare). In the single-cycle kinetics determination mode, compound titers were run at 45 μL/min. on each surface using a 2-minute association phase and a 30-minute dissociation phase in a buffer containing 50 mM Tris pH 7.5/150 mM NaCl/0.01% Tween 20/1 mM DTT/2% DMSO. Data were fitted using single-cycle small molecule kinetics (LMW) results. The equilibrium dissociation constants (K _D ) are given.

Different rapalogue pharmacology may be achieved in different cell or tissue types depending on 1) the relative abundance of FKBP homologs in such cells/tissues and 2) the specificity of binding to these different FKBP homologs (Mol. Cell Biol. (2013) 33:1357–1367 ).

results

In vitro activity of mTOR inhibitors was determined using pS6 IC ₅₀ (nM) in TSC1-/- MEF cells.

Compound IC50 (nM) Rapamycin 0.050 RAD001 0.050 Intermediate 1 0.092 Intermediate 3 0.0591 Intermediate 4 0.066 Intermediate 5 0.106 Example 1 2.4 Example 2 0.274 Example 3 1.45 Example 4 >500 Example 5 153 Example 6 0.170 Example 7 0.300 Example 8 0.328 Example 9 1.45 Example 10 0.557 Example 11 3.33 Example 12 0.374 Example 13 0.656 Example 14 7.77 Example 15 3.2 Example 16 0.645 Example 17 1.1 Example 18 1.55

IC50 values are calculated as the average of multiple assays.

Equilibrium dissociation constants ( _KD ) for FKBP12, FKBP51 and FKBP52.

Compound FKBP-
binding protein Average K _D (nM) RAD001 FKBP12:
FKBP51:
FKBP52: 500
811
1765 Example 1 FKBP12:
FKBP51:
FKBP52: 0.35
153
174 Example 2 FKBP12:
FKBP51:
FKBP52: 16
>10000
>10000 Example 6 FKBP12:
FKBP51:
FKBP52: 0.32
39
53 Example 7 FKBP12:
FKBP51:
FKBP52: 1.2
287
236 Example 10 FKBP12:
FKBP51:
FKBP52: 1.6
95
176 Example 14 FKBP12:
FKBP51:
FKBP52: 16
--
542 Example 16 FKBP12:
FKBP51:
FKBP52: 35
4064
4073

Pharmacokinetic profile of Example 2 in rats. Rats at 4 months of age were treated with a single po dose of Example 2 at 3 mg/kg and blood was collected at 0.25, 0.5, 1, 2, 4 and 7 hours post-dose. In this experiment, example 2 was made up in 15% polyethylene glycol (PEG) 300, 7.5% Solutol HS 15, 7.5% Cremophore EL in MilliQ water. The concentration values of Example 2 were measured using HPLC-MS ( Figure 3 ).

Comparative concentration values of example 2 and RAD001 in the blood and brain of rats.

Rats at 4 months of age were treated with po using Example 2 (formulated in 15% PEG 300, 7.5% Solutol HS 15, 7.5% Cremophore EL in MilliQ water) at 3, 10 and 30 mg/kg. In another experiment, 8-week-old rats were treated po with RAD001 (formulated as a microemulsion at 2% (w/w) and diluted to final concentration in MilliQ water) at 3, 10, and 30 mg/kg. Blood and brain tissues were collected 3 hours (h) and 24 h after treatment, and concentration values of compounds were determined. When administered at the same doses (3, 10 and 30 mg/kg), blood and brain concentration values for Example 2 were higher compared to those for RAD001 ( Figures 4A and 4B ).

To compare the bioavailability of Example 2 relative to RAD001 in rats, the compounds were formulated as solutions: Example 1 was formulated in 15% PEG300, 7.5% Solutol HS 15, 7.5% Cremophore EL in PBS and RAD001 was formulated in 10% PEG300, 10% Solutol HS 15, 10% Cremophore EL in PBS. The compounds were administered to 7-9 week old rats (N=3 per group) po at 3 mg/kg and intravenously (iv). at 1 mg/kg ( Figures 4C and 4D ). For example 2 and RAD001, respectively, bioavailability was 18% and 19%, terminal half-life after iv: 9.9 hours and 9.5 hours, clearance: 9 ml/min./kg and 32 ml/min./kg , and Vdss (volume of distribution at steady state): 4.4 l/kg and 18.8 l/kg.

Example 2 inhibits the mTORC1 pathway in rat liver. The ability of Example 2 to inhibit the mTORC1 pathway in vivo was determined in rats at 4 months of age ( Figures 5A - 5D ). A single dose of Example 2 at 3, 10 and 30 mg/kg produced significant S6 dephosphorylation (inactivation) in rat liver (compared to vehicle control) three hours post-dose ( Figures 5A and 5B ). S6 remained inactivated 24 hours post-dose at 10 (p=0.06 for time course) and 30 mg/kg ( Figures 5C and 5D ).

FKBP12 is required for the inhibitory effect of example 2, but not RAD001 . Amounts of phosphorylated S6K1 (Thr389) were measured in FKBP12 knockdown and FKBP12 knockout 293T WT cells treated with RAD001 or Example 2. S6K1 is a downstream target of mTORC1, and phosphorylation of its rapalog-sensitive site Thr389 is used as an effective indicator of mTORC1 activity. Lee, C. H., Inoki, K. and Guan, K. L. (2007). mTOR pathway as a target in tissue hypertrophy. Annu. Rev. Pharmacol. Toxicol. 47 , 443-467. In 293T WT cells, both RAD001 and example 2 were effective in inhibiting S6K1(Thr389) by ~80% ( Figure 6A ). In FKBP12 knockdown cells, Example 2 did not inhibit S6K1(Thr389) phosphorylation to the same level as RAD001: the maximum inhibition of S6K1(Thr389) by RAD001 was ~70%, whereas the inhibition achieved by Example 2 was ~40% % ( Figure 6B ). In FKBP12 knockout cells, Example 2 failed to provide inhibition of S6K1(Thr389) phosphorylation, while >60% inhibition was still achieved by RAD001 ( Figure 6C ). These results indicate that the pharmacological effects of Example 2 are limited to FKBP12. This specificity of Example 2 for FKBP12 may facilitate targeting of cells and tissues with relatively high levels of FKBP12 expression, while eliminating (or minimizing) off-target effects in tissues where FKBP12 expression levels are low.

Equivalents and scope of legal protection

In the claims, the singular forms of the singular may also include the plural unless otherwise indicated or otherwise obvious from the context. Claims or sections of the specification that include "or" between one or more members of the group are satisfied if one, more than one, or all members of the group are present in, used in, or otherwise related to the product or process, unless otherwise stated or otherwise obvious from the context. The present invention includes embodiments in which exactly one member of the group is present in, used in, or otherwise related to a given product or method. The present invention includes embodiments in which more than one or all members of the group are present in, used in, or otherwise related to a given product or method.

In addition, the present invention covers all variations, combinations and combinations in which one or more limitations, elements, claims and descriptive terms from one or more of the listed claims are presented in another claim. For example, any claim that is dependent on another claim may be modified to include one or more limitations found in any other claim that is dependent on the same independent claim. If the elements are represented as lists, e.g. in the Markush group format, each subgroup of elements is also open, and any element(s) can be removed from the group. It should be understood that in general, when the present invention or aspects of the present invention are stated to contain particular elements and/or features, some embodiments of the present invention or aspects of the present invention are or are substantially composed of such elements and/or features. It should also be noted that the terms “comprising” and “comprising” are intended to be open-ended and capable of including additional elements or steps. If ranges are given, endpoints are included. In addition, unless otherwise indicated or otherwise obvious from the context and understanding of one skilled in the art, values expressed as ranges can take on any specific value or subrange within the stated ranges in various embodiments of the present invention, up to a tenth units of the lower limit of the range, unless the context clearly requires otherwise.

This application makes reference to various issued patents, published patent applications, journal articles and other publications, all of which are incorporated herein by reference. If there is a discrepancy between any of the included literature and this description, the description will take precedence. Moreover, any specific embodiment of the present invention that is included in the prior art may be expressly excluded from any one or more claims. Since such embodiments are considered known to one skilled in the art, they may be excluded even if the exclusion is not expressly set forth herein. Any particular embodiment of the present invention may be omitted from any claim for any reason, regardless of whether it relates to the existence of prior art.

Those skilled in the art will identify, or be able to determine, using only standard experimentation, numerous equivalents to the specific embodiments described herein. It is intended that the scope of the embodiments of the present invention described herein is not limited to the description above, but instead is set forth in the accompanying claims. Those skilled in the art will appreciate that various changes and modifications can be made to this specification without departing from the spirit or scope of the present invention as defined in the following claims.

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LIST OF SEQUENCES

<110> NOVARTIS AG

<120> NEW RAPAMYCIN DERIVATIVES

<130> PAT057873-WO-PCT

<140> PCT/IB2018/057422

<141> 2018-09-25

<150> 62/563,312

<151> 2017-09-26

<160> 3

<170> PatentIn version 3.5

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<221> source

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Synthetic oligonucleotide"

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gcttctaaaa ctggaatgac

20

<210> 2

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic oligonucleotide"

<400> 2

gccactactc accgtctcct

20

<210> 3

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic 6xHis tag"

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His His His His His

15

<---

Claims (78)

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof (I), where R ₁ is selected from the group consisting of hydroxy, , , , And ; And R ₂ is selected from the group consisting of , , , where m equals 0; n is 1 or 2; And R ₃ represents hydrogen, C _1-6 alkyl or hydroxy-C _1-6 alkyl. 2. A compound according to claim 1 or a pharmaceutically acceptable salt thereof, where R ₃ represents hydrogen or C _1-6 alkyl. 3. A compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein R ₁ represents hydroxy. 4. Connection according to any one of paragraphs. 1-3 or a pharmaceutically acceptable salt thereof, where R ₂ represents and n is 1 or 2. 5. Connection according to any one of paragraphs. 1-4 or a pharmaceutically acceptable salt thereof, where R ₂ represents 6. Connection selected from: Structure
* Absolute stereochemical configuration for C16 is not specified
* Absolute stereochemical configuration for C16 is not specified
* Absolute stereochemical configuration for C16 is not specified
* Absolute stereochemical configuration for C16 is not specified
* Absolute stereochemical configuration for C16 is not specified
* Absolute stereochemical configuration for C16 is not specified
* Absolute stereochemical configuration for C16 is not specified
* Absolute stereochemical configuration for C16 is not specified
* Absolute stereochemical configuration for C16 is not specified
* Absolute stereochemical configuration for C16 is not specified .
* Absolute stereochemical configuration for C16 is not specified or a pharmaceutically acceptable salt thereof. 7. A compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein said compound is C16-(1,1-dioxidoisothiazolidin-2-yl)-C32-deoxyrapamycin. 8. A compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein said compound is (S)-C16-(1,1-dioxidoizothiazolidin-2-yl)-C32-deoxyrapamycin. 9. Connection according to any one of paragraphs. 1-5 or a pharmaceutically acceptable salt thereof, wherein said compound is (R)-C16-(1,1-dioxidoizothiazolidin-2-yl)-C32-deoxyrapamycin. 10. A compound according to claim 1 or a pharmaceutically acceptable salt thereof for use as a medicament for inhibiting mTOR activity. 11. A compound according to claim 1 or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of a disorder or disease mediated by the mTOR pathway. 12. A compound according to claim 1, or a pharmaceutically acceptable salt thereof, for use in the prevention or treatment of a disorder or disease mediated by the mTOR pathway selected from - acute or chronic organ or tissue transplant rejection; - proliferation and migration of smooth muscle cells, leading to thickening of the vascular intima, blockage of blood vessels, obstructive coronary atherosclerosis, restenosis; - autoimmune diseases and inflammatory conditions; - diseases associated with the treatment and prevention of asthma; - infections caused by fungi; - inflammation; - infections; - age-related diseases; - neurodegenerative diseases; - proliferative disorders, in particular cancer; - mitochondrial myopathy and mitochondrial stress and - treatable conditions that have been shown to increase the likelihood of age-related diseases, such as conditions in which there is an increase in the levels of cytokines that cause aging. 13. A compound according to claim 1, or a pharmaceutically acceptable salt thereof, for use in the prevention or treatment of a disorder or disease that involves a process of fibrosis or inflammation. 14. A compound for use according to claim 13, wherein the disorder is selected from liver and kidney disorders. 15. The compound for use according to claim 14, wherein the liver disorder is selected from liver fibrosis, which occurs in end-stage liver disease; liver cirrhosis; liver failure caused by toxicity; non-alcoholic steatohepatitis or NASH and alcoholic steatosis. 16. The compound for use according to claim 14, wherein the renal disorder is renal fibrosis that results from acute kidney injury. 17. A compound for use according to claim 14, wherein the renal disorder is a chronic kidney disorder. 18. A compound for use according to claim 14, wherein the renal disorder is diabetic nephropathy. 19. A compound according to claim 1, or a pharmaceutically acceptable salt thereof, for use in the prevention or treatment of an age-related disorder or disease mediated by the mTOR pathway selected from sarcopenia, osteoporosis, osteoarthritis, cognitive decline, tendon stiffness, cardiac dysfunction such as cardiac hypertrophy , and/or systolic and/or diastolic dysfunction, and/or hypertension, cardiac dysfunction that leads to decreased ejection fraction, aging of the immune system, Parkinson's disease, Alzheimer's disease, cancer, aging of the immune system leading to cancer due to decreased immune surveillance, infections due to decreased immune function, obesity, arthritis and type II diabetes, including diabetes-related complications such as kidney failure, blindness and neuropathy. 20. A compound according to claim 1 or a pharmaceutically acceptable salt thereof for use in the prevention or treatment of cancer mediated by the mTOR pathway. 21. A compound according to claim 1 or a pharmaceutically acceptable salt thereof, for use in the treatment of kidney cancer, renal cell carcinoma, colorectal cancer, uterine sarcoma, uterine endometrial cancer, endometrial cancer, breast cancer, ovarian cancer, cervical cancer, cancer stomach, fibrosarcoma, pancreatic cancer, liver cancer, melanoma, leukemia, multiple myeloma, nasopharyngeal cancer, prostate cancer, lung cancer, glioblastoma, bladder cancer, mesothelioma, head cancer, rhabdomyosarcoma, sarcoma, lymphoma or neck cancer. 22. Pharmaceutical composition for inhibiting mTOR activity, containing a therapeutically effective amount of a compound according to any one of claims. 1-9 or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers. 23. Use of a compound according to any one of paragraphs. 1-9 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 22 for the manufacture of a medicinal product intended for the prevention or treatment of a disorder or disease mediated by the mTOR pathway. 24. Use of the compound according to any one of paragraphs. 1-9 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 22 for the manufacture of a medicinal product intended for the prevention or treatment of a disorder or disease mediated by the mTOR pathway selected from: - acute or chronic organ or tissue transplant rejection; - graft-related vasculopathies; - proliferation and migration of smooth muscle cells, leading to thickening of the vascular intima, blockage of blood vessels, obstructive coronary atherosclerosis, restenosis; - autoimmune diseases and inflammatory conditions; - diseases associated with the treatment and prevention of asthma; - infections caused by fungi; - inflammation; - infections; - age-related diseases; - neurodegenerative diseases; - proliferative disorders, in particular cancer; - mitochondrial myopathy and mitochondrial stress and - treatable conditions that have been shown to increase the likelihood of age-related diseases, such as conditions in which there is an increase in the levels of cytokines that cause aging. 25. Use of a compound according to any one of paragraphs. 1-9 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 22 for the manufacture of a medicinal product intended for the prevention or treatment of a disorder or disease mediated by the mTOR pathway, which involves the process of fibrosis or inflammation. 26. Use according to claim 25, where the disorder is selected from liver and kidney disorders. 27. Use according to claim 26, wherein the liver disorder is selected from liver fibrosis, which occurs in the end stage of liver disease; liver cirrhosis; liver failure caused by toxicity; non-alcoholic steatohepatitis or NASH and alcoholic steatosis. 28. Use according to claim 26, wherein the renal disorder is renal fibrosis that occurs as a result of acute kidney injury. 29. Use according to claim 26, wherein the renal disorder is a chronic renal disorder. 30. Use according to claim 26, wherein the renal disorder is diabetic nephropathy. 31. Use of a compound according to any one of paragraphs. 1-9 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 22 for the manufacture of a medicinal product intended for the prevention or treatment of an age-related disorder or disease mediated by the mTOR pathway selected from sarcopenia, osteoporosis, osteoarthritis, cognitive decline, tendon stiffness, dysfunction heart disease, such as cardiac hypertrophy, and/or systolic and/or diastolic dysfunction, and/or hypertension, cardiac dysfunction leading to decreased ejection fraction, aging of the immune system, Parkinson's disease, Alzheimer's disease, cancer, aging of the immune system leading to cancer due to decreased immune surveillance, infections due to decreased immune function, obesity, arthritis and type II diabetes, including diabetes-related complications such as kidney failure, blindness and neuropathy. 32. Use of a compound according to any one of paragraphs. 1-9 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 22 for the manufacture of a medicinal product intended for the prevention or treatment of cancer mediated by the mTOR pathway. 33. Use of a compound according to any one of paragraphs. 1-9 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 22 for the manufacture of a medicament for the treatment of a disorder or disease mediated by the mTOR pathway selected from kidney cancer, renal cell carcinoma, colorectal cancer, uterine sarcoma, uterine endometrial cancer , endometrial cancer, breast cancer, ovarian cancer, cervical cancer, stomach cancer, fibrosarcoma, pancreatic cancer, liver cancer, melanoma, leukemia, multiple myeloma, nasopharyngeal cancer, prostate cancer, lung cancer, glioblastoma, bladder cancer, mesothelioma, head cancer, rhabdomyosarcoma, sarcoma, lymphoma or neck cancer.