PRMT5 inhibitors and their uses

Abstract

This disclosure describes compounds of Formulas (I)-(V) and methods for making them. The compounds of this disclosure are useful as inhibitors of PRMT5 activity and in methods for treating cancer and other diseases. [Formula 1] TIFF2022521818000238.tif37130

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Patent Application No. 62 / 823,374, filed March 25, 2019, and U.S. Provisional Patent Application No. 62 / 874,773, filed July 16, 2019, each of which is incorporated herein by reference in its entirety. [Background technology]

[0002] PRMT5 is one of the major symmetric arginine methyltransferases and is involved in various cellular processes, so increased protein expression of PRMT5 is associated with its tumorigenicity. Elevated PRMT5-MEP50 protein levels and cytoplasmic accumulation of this complex are also both associated with cancer tumorigenicity and correlated with poor clinical outcomes. New small molecules (i.e., compounds) that inhibit PRMT5 activity are needed to treat a wide range of cancers and other diseases. Summary of the Invention

[0003] In certain embodiments, the present invention provides (a) Formula I:

[0004] [ka] or a pharma- ceutically acceptable salt thereof. [In the formula,

[0005] [ka] is a double bond or a single bond, Ar is an optionally substituted aryl or heteroaryl, where one, two or three optional substituents are halogen, hydroxyl, cyano, -C(O)NH, -C(O)N(R a )2, -C(O)O(R a ), -C(O)R a , -N(R a)2, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl, optionally substituted C1-C4 alkyl, and optionally substituted C2-C6 alkenyl; R1 is H, halogen, C1-C4 alkyl, C2-C4 alkenyl, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl or N(R a )2, X1 and X2 are each independently CH or N; Y1 is (CH2) n or C(O), with the proviso that when Y2 or Y3 is C(O), then Y1 cannot be C(O); Y2 is CH2, C(O), S, SO2, O or NR a However, when Y1 or Y3 is C(O), Y2 cannot be C(O);

[0006] [ka] is a single bond, Y3 is CH2 or C(O), provided When Y1 or Y2 is C(O), Y3 cannot be C(O);

[0007] [ka] is a single bond, or Y2 and Y3 are CH;

[0008] [ka] is a double bond, Z is CH2, O, S or NH; R a is independently, at each occurrence, H, C1-C4 alkyl, or C3-C6 cycloalkyl; m is 0, 1, 2 or 3; n is 1 or 2; or (b) Formula II:

[0009] [ka] or a pharma- ceutically acceptable salt thereof. [In the formula,

[0010] [ka] is, independently in each occurrence, a double bond or a single bond; R1 is H, halogen, C1-C4 alkyl, C2-C4 alkenyl, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl or N(R a )2, R2 is H, halogen, hydroxyl, cyano, -C(O)NH2, -C(O)N(R a )2, -C(O)O(R a ), -C(O)R a , -N(R a )2, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl, optionally substituted C1-C4 alkyl or optionally substituted C2-C6 alkenyl; W is C(R b ) or C(O), provided that when W is C(O), V is NH, and

[0011] [ka] is a single bond, V is CH, NH, or N, provided that when V is CH or N, W is C(R b ) between W and V

[0012] [ka] is a double bond, X1 and X2 are each independently CH or N; Y1 is (CH2) n or C(O), with the proviso that when Y2 or Y3 is C(O), then Y1 cannot be C(O); Y2 is CH2, C(O), S, SO2, O or NR a However, when Y1 or Y3 is C(O), Y2 cannot be C(O); Between Y2 and Y3

[0013] [ka] is a single bond, Y3 is CH2 or C(O), provided When Y1 or Y2 is C(O), Y3 cannot be C(O); Between Y2 and Y3

[0014] [ka] is a single bond, or Y2 and Y3 are CH;

[0015] [ka] is a double bond, Z is CH2, O, S or NH; R a is independently, at each occurrence, H, C1-C4 alkyl, or C3-C6 cycloalkyl; R b is H, halogen, hydroxyl, cyano, -C(O)NH2, -C(O)N(R a )2, -C(O)O(R a ), -C(O)R a , -N(R a )2, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl, optionally substituted C1-C4 alkyl or optionally substituted C2-C6 alkenyl; m is 0, 1, 2 or 3; n is 1 or 2; or (c) Formula III:

[0016] [ka] or a pharma- ceutically acceptable salt thereof. [In the formula, Ar is an optionally substituted aryl or heteroaryl, where one, two or three optional substituents are halogen, hydroxyl, cyano, -C(O)NH, -C(O)N(R a )2, -C(O)O(R a ), -C(O)R a , -N(R a )2, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl, optionally substituted C1-C4 alkyl, and optionally substituted C2-C6 alkenyl; R1 is H, halogen, C1-C4 alkyl, C2-C4 alkenyl, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl or N(R a )2, X1 and X2 are each independently CH or N; Y2 is CH2, S, O or N(R a ) and Z is CH2, O, S or NH; R a is independently, at each occurrence, H, C1-C4 alkyl, or C3-C6 cycloalkyl; m is 0, 1, 2 or 3; or (d) Formula IV:

[0017] [ka] or a pharma- ceutically acceptable salt thereof. [In the formula, R1 is H, halogen, C1-C4 alkyl, C2-C4 alkenyl, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl or N(R a )2, R2 is H, halogen, hydroxyl, cyano, -C(O)NH2, -C(O)N(R a )2, -C(O)O(R a ), -C(O)R a , -N(R a )2, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl, optionally substituted C1-C4 alkyl or optionally substituted C2-C6 alkenyl; X1 and X2 are each independently CH or N; V is CH or N; Y2 is CH2, S, O or N(R a ) and Z is CH2, O, S or NH; R a is independently, at each occurrence, H, C1-C4 alkyl, or C3-C6 cycloalkyl; R b is H, halogen, hydroxyl, cyano, -C(O)NH2, -C(O)N(R a )2, -C(O)O(R a ), -C(O)R a , -N(R a )2, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl, optionally substituted C1-C4 alkyl or optionally substituted C2-C6 alkenyl; or (e) Formula V:

[0018] [ka] or a pharma- ceutically acceptable salt thereof. [In the formula, R1 is H, halogen, C1-C4 alkyl, C2-C4 alkenyl, -O(R a), C3-C6 cycloalkyl, C3-C7 heterocyclyl or N(R a )2, R2 is H, halogen, hydroxyl, cyano, -C(O)NH2, -C(O)N(R a )2, -C(O)O(R a ), -C(O)R a , -N(R a )2, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl, optionally substituted C1-C4 alkyl or optionally substituted C2-C6 alkenyl; X2 is CH or N; Y2 is CH2, S, O or N(R a ), and R a is independently, at each occurrence, H, C1-C4 alkyl, or C3-C6 cycloalkyl. Regarding. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0019] definition Unless otherwise specified herein, scientific and technical terms used in this application shall have the meanings commonly understood by those of ordinary skill in the art. In general, the nomenclature used in connection with, and techniques of, chemistry, cell and tissue culture, molecular biology, cell and cancer biology, neurobiology, neurochemistry, virology, immunology, microbiology, pharmacology, genetics, and protein and nucleic acid chemistry described herein are those well known and commonly used in the art.

[0020] The methods and techniques of the present disclosure, unless otherwise indicated, are generally performed according to conventional methods well known in the art and as described in the various general and more specific references cited and discussed throughout this specification. See, for example, "Principles of Neural Science", McGraw-Hill Medical, New York, NY (2000); Motulsky, "Intuitive Biostatistics", Oxford University Press, Inc. (1995); Lodish et al., "Molecular Cell Biology, 4th ed.", WH Freeman & Co., New York (2000); Griffiths et al., "Introduction to Genetic Analysis, 7th ed.", WH Freeman & Co., NY (1999); and Gilbert et al., "Developmental Biology, 6th ed.", Sinauer Associates, Inc., Sunderland, MA (2000).

[0021] Chemical terms used herein, unless otherwise defined herein, are used according to conventional usage in the art as exemplified by "The McGraw-Hill Dictionary of Chemical Terms", Parker S., Ed., McGraw-Hill, San Francisco, CA (1985).

[0022] All of these and any other publications, patents and published patent applications referenced herein are specifically incorporated herein by reference. In case of conflict, the present specification, including its specific definitions, will control.

[0023] "Patient," "subject," or "individual" are used interchangeably and refer to either a human or non-human animal. These terms include mammals, such as humans, primates, livestock animals (including cows, pigs, etc.), pet animals (e.g., dogs, cats, etc.), and rodents (e.g., mice and rats).

[0024] "Treating" a condition or patient refers to taking steps to obtain beneficial or desired results, including clinical results. As used herein, and as is well understood in the art, "treatment" is an approach to obtain beneficial or desired results, including clinical results. Beneficial or desired clinical results may include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, whether detectable or undetectable, reduction in the extent of disease, stabilization of the disease state (i.e., not worsening), prevention of disease spread, delay or slowing of disease progression, improvement or palliation of the disease state, and remission (whether partial or total). "Treatment" may also mean prolonging survival compared to expected survival if not treated.

[0025] The term "preventing" is art-recognized and well understood in the art when used in connection with a condition, e.g., local recurrence (e.g., pain), a disease, e.g., cancer, a complex syndrome, e.g., heart failure, or any other medical condition, and includes administration of a composition that reduces the frequency or delays the onset of symptoms of a medical condition in a subject compared to subjects not receiving the composition. Thus, prevention of cancer includes, for example, reducing the number of detectable cancerous growths in a population of patients receiving a prophylactic treatment compared to an untreated control population by a statistically and / or clinically significant amount, and / or delaying the appearance of detectable cancerous growths in a treated population compared to an untreated control population.

[0026] "Administering" or "administration" of a substance, compound or agent to a subject can be performed using one of a variety of methods known to those skilled in the art. For example, the compound or agent can be administered intravenously, intraarterially, intradermally, intramuscularly, intraperitoneally, subcutaneously, ocularly, sublingually, orally (by ingestion), intranasally (by inhalation), intraspinally, intracerebrally, and transdermally (e.g., by absorption through the skin's channels). The compound or agent can also be suitably introduced by rechargeable or biodegradable polymeric or other devices, such as patches and pumps, or formulations that provide sustained, slow or controlled release of the compound or agent. Administration can also be performed, for example, once, multiple times, and / or over one or more extended periods of time.

[0027] Suitable methods for administering a substance, compound or agent to a subject will also depend, for example, on the age and / or physical condition of the subject, and the chemical and biological properties of the compound or agent (e.g., solubility, digestibility, bioavailability, stability and toxicity). In some embodiments, the compound or agent is orally administered to the subject, for example, by ingestion. In some embodiments, the orally administered compound or agent is in a sustained or timed release formulation or is administered using such a sustained or timed release device.

[0028] An "alkyl" group or "alkane" is a straight-chain or branched non-aromatic hydrocarbon that is fully saturated. Typically, a straight-chain or branched alkyl group has from 1 to about 20 carbon atoms, preferably from 1 to about 10 carbon atoms, or more preferably from 1 to about 3 carbon atoms, unless otherwise specified. Examples of straight-chain and branched alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, tert-butyl, pentyl, hexyl, pentyl, and octyl. C1-C6 straight-chain or branched alkyl groups are also referred to as "lower alkyl" groups.

[0029] Furthermore, the term "alkyl" (or "lower alkyl"), as used throughout the specification, examples, and claims, is intended to include both "unsubstituted alkyl" and "substituted alkyl," the latter of which refers to alkyl moieties having substituents replacing a hydrogen on one or more carbons of the hydrocarbon backbone. Such substituents, unless otherwise specified, include, for example, halogen (e.g., fluoro), hydroxyl, oxo, carbonyl (e.g., carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (e.g., thioester, thioacetate, or thioformate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, heterocyclyl, aralkyl, or aromatic or heteroaromatic moieties. In preferred embodiments, the substituents on the substituted alkyl are selected from C1-C6 alkyl, C3-C6 cycloalkyl, halogen, carbonyl, cyano, or hydroxyl. In more preferred embodiments, the substituents on the substituted alkyl are selected from fluoro, carbonyl, cyano, or hydroxyl. It will be understood by those skilled in the art that the moieties substituted on the hydrocarbon chain can themselves be substituted, where appropriate. For example, substituents on the substituted alkyl include substituted and unsubstituted amino, azido, imino, amido, phosphoryl (including phosphonates and phosphinates), sulfonyl (including sulfates, sulfonamides, sulfamoyl, and sulfonates), and silyl groups, as well as ethers, alkylthio, carbonyl (including ketones, aldehydes, carboxylates, and esters), -CF3, -CN, and the like. Exemplary substituted alkyls are described below. Cycloalkyls can be further substituted with alkyls, alkenyls, alkoxy, alkylthios, aminoalkyls, carbonyl-substituted alkyls, -CF3, -CN, and the like.

[0030] Additionally, the term "alkyl," as used throughout the specification, examples, and claims, is intended to be interchangeable with the term "alkylene" (i.e., an alkyl diradical or divalent alkyl moiety) where valency requires.

[0031] The term "alkenyl" as used herein refers to an aliphatic group containing at least one double bond, and is intended to include both "unsubstituted alkenyl" and "substituted alkenyl", the latter of which refers to an alkenyl moiety having a substituent replacing a hydrogen on one or more carbons of the alkenyl group. Such substituents may be present on one or more carbons that are or are not included in one or more double bonds. Furthermore, such substituents include all of those contemplated for alkyl groups, as described below, except where stability prohibits their use. For example, substitution of alkenyl groups with one or more alkyl, carbocyclyl, aryl, heterocyclyl, or heteroaryl groups is contemplated.

[0032] The term "alkynyl" as used herein refers to an aliphatic group containing at least one triple bond, and is intended to include both "unsubstituted alkynyl" and "substituted alkynyl", the latter of which refers to an alkynyl moiety having a substituent replacing a hydrogen on one or more carbons of the alkynyl group. Such a substituent may be present on one or more carbons that are or are not included in one or more triple bonds. Furthermore, such substituents include all of those contemplated for alkyl groups, as described above, except where stability prohibits their use. For example, substitution of alkynyl groups with one or more alkyl, carbocyclyl, aryl, heterocyclyl, or heteroaryl groups is contemplated.

[0033] The term “C x ~C y " when used in conjunction with a chemical moiety, such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, is meant to include groups that contain x to y carbons in the chain. For example, the term "C x ~Cy "Alkyl" refers to substituted or unsubstituted saturated hydrocarbon groups, including straight chain alkyl and branched alkyl groups, containing x to y carbons in the chain, including haloalkyl groups. Preferred haloalkyl groups include trifluoromethyl, difluoromethyl, 2,2,2-trifluoroethyl and pentafluoroethyl. CO alkyl represents a hydrogen when the group is in a terminal position and a bond when the group is internal. The term "C2-C y Alkenyl" and "C2-C y "Alkynyl" refers to substituted or unsubstituted unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but which contain at least one double or triple bond respectively.

[0034] The term "aryl" as used herein includes substituted or unsubstituted monocyclic aromatic groups in which each atom of the ring is carbon. Preferably, the ring is 6-10 membered, more preferably 6 membered. The term "aryl" also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjacent rings, where at least one of the rings is aromatic, and the other cyclic rings can be, for example, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl and / or heterocyclyl. Aryl groups include benzene, naphthalene, phenanthrene, aniline, and the like.

[0035] The term "carbocycle" refers to a saturated or unsaturated ring in which each atom of the ring is carbon. The term carbocycle includes both aromatic and non-aromatic carbocycles. Non-aromatic carbocycles include both cycloalkyl and cycloalkenyl rings. "Carbocycle" includes 5- to 7-membered monocyclic rings and 8- to 12-membered bicyclic rings. Each ring of a bicyclic carbocycle can be selected from saturated, unsaturated, and aromatic rings. Carbocycles include bicyclic molecules in which one, two, or three or more atoms are shared between the two rings. Carbocycles include bicyclic molecules in which one, two, or three or more atoms are shared between the two rings. The term "fused carbocycle" refers to a bicyclic carbocycle in which each of the rings shares two adjacent atoms with the other ring. Each ring of a fused carbocycle can be selected from saturated, unsaturated, and aromatic rings. In exemplary embodiments, an aromatic ring, such as phenyl, may be fused to a saturated or unsaturated ring, such as cyclohexane, cyclopentane, or cyclohexene. Any combination of saturated, unsaturated, and aromatic bicyclic rings is included in the definition of carbocycle, if valences permit. Exemplary "carbocycles" include cyclopentane, cyclohexane, bicyclo[2.2.1]heptane, 1,5-cyclooctadiene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]oct-3-ene, naphthalene, and adamantane. Exemplary fused carbocycles include decalin, naphthalene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]octane, 4,5,6,7-tetrahydro-1H-indene, and bicyclo[4.1.0]hept-3-ene. A "carbocycle" may be optionally substituted at any one or more positions capable of carrying a hydrogen atom.

[0036] A "cycloalkyl" group is a fully saturated cyclic hydrocarbon. "Cycloalkyl" includes monocyclic and bicyclic rings. Typically, a monocyclic cycloalkyl group has 3 to about 10 carbon atoms, 3 to 8 carbon atoms, or more typically 3 to 6 carbon atoms, unless otherwise specified. The second ring of a bicyclic cycloalkyl can be selected from saturated, unsaturated, and aromatic rings. Cycloalkyl includes bicyclic molecules in which one, two, or three or more atoms are shared between the two rings (e.g., fused bicyclic compounds, bridged bicyclic compounds, and spirocyclic compounds).

[0037] The term "fused bicyclic compound" refers to a bicyclic molecule in which two rings share two adjacent atoms. In other words, these rings share one covalent bond, i.e., the so-called bridgehead atoms are directly bonded (e.g., α-thujene and decalin). For example, in fused cycloalkyl, each of the rings shares two adjacent atoms with the other ring, and the second ring of the fused bicyclic cycloalkyl can be selected from saturated, unsaturated and aromatic rings.

[0038] The term "spirocyclic compound" refers to a bicyclic molecule in which the two rings have only one single atom in common, the spiroatom.

[0039] The terms "heteroaryl" and "hetaryl" include substituted or unsubstituted aromatic monocyclic structures (preferably 5-7 membered rings, more preferably 5-6 membered rings) that contain at least one heteroatom, preferably 1-4 heteroatoms, more preferably 1 or 2 heteroatoms. The terms "heteroaryl" and "hetaryl" also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjacent rings, where at least one of the rings is heteroaromatic, and the other cyclic rings can be, for example, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine and pyrimidine, quinoline, quinoxaline, naphthyridine, and the like.

[0040] The term "heteroatom" as used herein means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen and sulfur.

[0041] The terms "heterocyclyl", "heterocycle", and "heterocyclic" refer to substituted or unsubstituted non-aromatic ring structures, preferably 3-10 membered rings, preferably 3-7 membered rings, more preferably 5-6 membered rings, and in some cases most preferably 5 membered rings, and in other cases most preferably 6 membered rings, whose ring structures contain at least one heteroatom, preferably 1-4 heteroatoms, more preferably 1 or 2 heteroatoms. The terms "heterocyclyl" and "heterocyclic" also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjacent rings, where at least one of the rings is heterocyclic, for example, the other cyclic rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl and / or heterocyclyl. Heterocyclyl groups include, for example, pyrrolidine, piperidine, piperazine, pyrrolidine, tetrahydropyran, tetrahydrofuran, morpholine, lactones, lactams, oxazolines, imidazolines, and the like.

[0042] The terms "halo" and "halogen" are used interchangeably herein to mean halogen, including chloro, fluoro, bromo, and iodo.

[0043] The term "substituted" refers to a moiety having a substituent replacing a hydrogen on one or more carbons of the backbone. It will be understood that "substituted" or "substituted with" includes the implicit proviso that such substitution is subject to the allowed valence of the substituted atom and substituent, as well as that the substitution results in a stable compound that does not spontaneously undergo transformation, for example, by rearrangement, cyclization, elimination, and the like. For appropriate organic compounds, the substitutions may be one or more and may be the same or different.

[0044] The phrase "pharmacologically acceptable" is art-recognized. In certain embodiments, this term includes compositions, excipients, adjuvants, polymers, and other materials and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without undue toxicity, irritation, allergic response, or other problem or complication, and are commensurate with a reasonable benefit / risk ratio.

[0045] "Pharmaceutically acceptable salt" or "salt" is used herein to refer to acid addition salts or base addition salts that are suitable or compatible with the treatment of patients.

[0046] The term "pharmaceutically acceptable acid addition salts" as used herein means any non-toxic organic or inorganic salt of any base compound disclosed herein. Exemplary inorganic acids that form suitable salts include hydrochloric acid, hydrobromic acid, sulfuric acid, and phosphoric acid, as well as metal salts such as sodium monohydrogen orthophosphate and potassium hydrogen sulfate. Exemplary organic acids that form suitable salts include mono-, di-, and tricarboxylic acids such as glycolic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, malic acid, tartaric acid, citric acid, ascorbic acid, maleic acid, benzoic acid, phenylacetic acid, cinnamic acid, and salicylic acid, as well as sulfonic acids such as p-toluenesulfonic acid and methanesulfonic acid. Either mono- or di-acid salts can be formed, and such salts can exist in either hydrated, solvated, or substantially anhydrous form. In general, acid addition salts of the compounds disclosed herein are more soluble in water and various hydrophilic organic solvents, and generally exhibit higher melting points compared to their free base forms. The selection of an appropriate salt will be known to one of skill in the art. Other pharmaceutically unacceptable salts, such as oxalates, may be used, for example, in the isolation of compounds of the invention for laboratory use or for subsequent conversion to a pharmaceutically acceptable acid addition salt.

[0047] The term "pharmaceutically acceptable base addition salt" as used herein means any non-toxic organic or inorganic base addition salt of any acid compound of the present invention, or any of its intermediates. Exemplary inorganic bases that form suitable salts include lithium hydroxide, sodium hydroxide, potassium hydroxide, calcium hydroxide, magnesium hydroxide, or barium hydroxide. Exemplary organic bases that form suitable salts include aliphatic, alicyclic, or aromatic organic amines, such as methylamine, trimethylamine, and picoline, or ammonia. The selection of appropriate salts will be known to those skilled in the art.

[0048] Many of the compounds useful in the methods and compositions of the present disclosure have at least one stereocenter in their structure. This stereocenter may exist in the R or S configuration, and the R and S designations are used in accordance with the rules set forth in Pure Appl. Chem. (1976), 45, 11-30. The present disclosure contemplates all stereoisomeric forms, such as enantiomeric and diastereoisomeric forms (including all possible mixtures of stereoisomers), of the compounds, salts, prodrugs, or mixtures thereof. See, for example, WO 01 / 062726. If the absolute stereochemistry of a stereocenter has not been determined, the stereochemistry is

[0049] [ka] In other words, the structural formula

[0050] [ka] [In the formula, R 10 , R 11 , R 12 , and R 13 are different from each other] Compounds having the formula: have either the R or S configuration. Where the stereochemistry at a stereocenter is not indicated in the structure as drawn herein, the drawn structure is intended to encompass racemic or other mixtures of enantiomeric or diastereomeric forms, and isolated enantiomers of any stereochemical configuration.

[0051] Furthermore, certain compounds containing alkenyl groups can exist as Z (zusammen) or E (entgegen) isomers, and in each case the present disclosure includes both mixtures and the separate individual isomers.

[0052] Some of the compounds may also exist in tautomeric forms. Such forms, although not explicitly shown in the formulae depicted herein, are intended to be included within the scope of the present disclosure.

[0053] "Prodrug" or "Pharmaceutically acceptable prodrug" refers to a compound that is metabolized, e.g., hydrolyzed or oxidized, in the host after administration to form a compound of the present disclosure (e.g., a compound of the present invention). Typical examples of prodrugs include compounds that have a biologically labile or cleavable (protecting) group on a functional moiety of an active compound. Prodrugs include compounds that can be oxidized, reduced, aminated, deaminated, hydroxylated, dehydroxylated, hydrolyzed, dehydrolyzed, alkylated, dealkylated, acylated, deacylated, phosphorylated, or dephosphorylated to generate an active compound. Examples of prodrugs that use esters or phosphoramidates as biologically labile or cleavable (protecting) groups are disclosed in U.S. Patent Nos. 6,875,751, 7,585,851, and 7,964,580, the disclosures of which are incorporated herein by reference. The prodrugs of the present disclosure are metabolized to generate a compound of the present invention or a pharmaceutically acceptable salt thereof. The present disclosure includes within its scope prodrugs of the compounds described herein. Conventional procedures for the selection and preparation of suitable prodrugs are described, for example, in “Design of Prodrugs” Ed. H. Bundgaard, Elsevier, 1985.

[0054] Exemplary Compounds In certain embodiments, the present invention provides a compound of formula I:

[0055] [ka] or a pharma- ceutically acceptable salt thereof. [In the formula,

[0056] [ka] is a double bond or a single bond, Ar is an optionally substituted aryl or heteroaryl, where one, two or three optional substituents are halogen, hydroxyl, cyano, -C(O)NH, -C(O)N(Ra )2, -C(O)O(R a ), -C(O)R a , -N(R a )2, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl, optionally substituted C1-C4 alkyl, and optionally substituted C2-C6 alkenyl; R1 is H, halogen, C1-C4 alkyl, C2-C4 alkenyl, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl or N(R a )2, X1 and X2 are each independently CH or N; Y1 is (CH2) n or C(O), with the proviso that when Y2 or Y3 is C(O), then Y1 cannot be C(O); Y2 is CH2, C(O), S, SO2, O or NR a However, when Y1 or Y3 is C(O), Y2 cannot be C(O);

[0057] [ka] is a single bond, Y3 is CH2 or C(O), provided When Y1 or Y2 is C(O), Y3 cannot be C(O);

[0058] [ka] is a single bond, or Y2 and Y3 are CH;

[0059] [ka] is a double bond, Z is CH2, O, S or NH; R ais independently, at each occurrence, H, C1-C4 alkyl, or C3-C6 cycloalkyl; m is 0, 1, 2 or 3; n is 1 or 2. Regarding.

[0060] In other embodiments, Ar is an optionally substituted aryl or heteroaryl, wherein one, two or three optional substituents are F, Cl, Br, methyl, N(R a )2 and O(R a ), R1 is independently selected from H, F, Cl, Br or N(R a )2, Z is CH2, O or NH, R a is independently at each occurrence H or methyl, and m is 0 or 1.

[0061] In certain embodiments, the compound of formula I has formula Ia:

[0062] [ka] or a pharma- ceutically acceptable salt thereof. Alternatively, the compound of formula I has the structure of formula Ib:

[0063] [ka] or a pharma- ceutically acceptable salt thereof.

[0064] In certain embodiments, the compound of formula I is Formula II:

[0065] [ka] or a pharma- ceutically acceptable salt thereof [In the formula,

[0066] [ka] is, independently in each occurrence, a double bond or a single bond; R1 is H, halogen, C1-C4 alkyl, C2-C4 alkenyl, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl or N(R a )2, R2 is H, halogen, hydroxyl, cyano, -C(O)NH2, -C(O)N(R a )2, -C(O)O(R a ), -C(O)R a , -N(R a )2, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl, optionally substituted C1-C4 alkyl or optionally substituted C2-C6 alkenyl; W is C(R b ) or C(O), provided that when W is C(O), V is NH, and

[0067] [ka] is a single bond, V is CH, NH, or N, provided that when V is CH or N, W is C(R b ) between W and V

[0068] [ka] is a double bond, X1 and X2 are each independently CH or N; Y1 is (CH2) n or C(O), with the proviso that when Y2 or Y3 is C(O), then Y1 cannot be C(O); Y2 is CH2, C(O), S, SO2, O or NR a However, when Y1 or Y3 is C(O), Y2 cannot be C(O); Between Y2 and Y3

[0069] [ka] is a single bond, Y3 is CH2 or C(O), provided When Y1 or Y2 is C(O), Y3 cannot be C(O); Between Y2 and Y3

[0070] [ka] is a single bond, or Y2 and Y3 are CH;

[0071] [ka] is a double bond, Z is CH2, O, S or NH; R a is independently, at each occurrence, H, C1-C4 alkyl, or C3-C6 cycloalkyl; R b is H, halogen, hydroxyl, cyano, -C(O)NH2, -C(O)N(R a )2, -C(O)O(R a ), -C(O)R a , -N(R a )2, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl, optionally substituted C1-C4 alkyl or optionally substituted C2-C6 alkenyl; m is 0, 1, 2 or 3; n is 1 or 2. It has the structure:

[0072] In other embodiments, R1 and R2 are each independently H, F, Cl, Br, or N(R a )2, Z is CH2, O or NH, R a is independently, at each occurrence, H or methyl; R b is H, N(R a) 2, F, Cl or Br, and m is 0 or 1.

[0073] In certain embodiments, the compound of formula II has formula IIa:

[0074] [ka] or a pharma- ceutically acceptable salt thereof. Alternatively, the compound of formula II has the structure of formula IIb:

[0075] [ka] or a pharma- ceutically acceptable salt thereof.

[0076] In certain embodiments, the compound of formula I has formula III:

[0077] [ka] or a pharma- ceutically acceptable salt thereof [In the formula, Ar is an optionally substituted aryl or heteroaryl, where one, two or three optional substituents are halogen, hydroxyl, cyano, -C(O)NH, -C(O)N(R a )2, -C(O)O(R a ), -C(O)R a , -N(R a )2, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl, optionally substituted C1-C4 alkyl, and optionally substituted C2-C6 alkenyl; R1 is H, halogen, C1-C4 alkyl, C2-C4 alkenyl, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl or N(R a )2, X1 and X2 are each independently CH or N; Y2 is CH2, S, O or N(R a) and Z is CH2, O, S or NH; R a is independently, at each occurrence, H, C1-C4 alkyl, or C3-C6 cycloalkyl; m is 0, 1, 2 or 3. It has the structure:

[0078] In certain embodiments, the compound of formula III has formula IIIa:

[0079] [ka] or a pharma- ceutically acceptable salt thereof. In certain embodiments of the compound of formula IIIa, m is 0. Alternatively, the compound of formula III has the structure of formula IIIb:

[0080] [ka] or a pharma- ceutically acceptable salt thereof. In certain embodiments of the compound of formula IIIb, m is 0.

[0081] In certain embodiments, the compound of formula I has formula IV:

[0082] [ka] or a pharma- ceutically acceptable salt thereof [In the formula, R1 is H, halogen, C1-C4 alkyl, C2-C4 alkenyl, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl or N(R a )2, R2 is H, halogen, hydroxyl, cyano, -C(O)NH2, -C(O)N(R a )2, -C(O)O(R a ), -C(O)R a , -N(R a )2, -O(R a), C3-C6 cycloalkyl, C3-C7 heterocyclyl, optionally substituted C1-C4 alkyl or optionally substituted C2-C6 alkenyl; X1 and X2 are each independently CH or N; V is CH or N; Y2 is CH2, S, O or N(R a ) and Z is CH2, O, S or NH; R a is independently, at each occurrence, H, C1-C4 alkyl, or C3-C6 cycloalkyl; R b is H, halogen, hydroxyl, cyano, -C(O)NH2, -C(O)N(R a )2, -C(O)O(R a ), -C(O)R a , -N(R a )2, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl, optionally substituted C1-C4 alkyl or optionally substituted C2-C6 alkenyl. It has the structure:

[0083] In other embodiments, R1 and R2 are each independently H, F, Cl, Br, or N(R a )2, Z is CH2, O or NH, R a is independently, at each occurrence, H or methyl; R b is H, N(R a ) 2, F, Cl or Br.

[0084] In certain embodiments, the compound of formula IV has formula IVa:

[0085] [ka] or a pharma- ceutically acceptable salt thereof. Alternatively, the compound of formula IV has the structure of formula IVb:

[0086] [ka] or a pharma- ceutically acceptable salt thereof.

[0087] In certain embodiments, the compound of formula I has formula V:

[0088] [ka] or a pharma- ceutically acceptable salt thereof [In the formula, R1 is H, halogen, C1-C4 alkyl, C2-C4 alkenyl, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl or N(R a )2, R2 is H, halogen, hydroxyl, cyano, -C(O)NH2, -C(O)N(R a )2, -C(O)O(R a ), -C(O)R a , -N(R a )2, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl, optionally substituted C1-C4 alkyl or optionally substituted C2-C6 alkenyl; X2 is CH or N; Y2 is CH2, S, O or N(R a ) and R a is independently, at each occurrence, H, C1-C4 alkyl, or C3-C6 cycloalkyl. It has the structure:

[0089] In certain embodiments, the compound of formula V has formula Va:

[0090] [ka] or a pharma- ceutically acceptable salt thereof. Alternatively, the compound of formula V has the structure of formula Vb:

[0091] [ka] or a pharma- ceutically acceptable salt thereof.

[0092] In some embodiments, the invention relates to a compound having the structure of Formula V, Va, or Vb, wherein the compound has a PRMT5 IC of about 100 nM or less. 50 has.

[0093] In further embodiments, the compound of Formula V, Va, or Vb has an average percent maximal efficacy against a drug-resistant cell line of Table 4 of about 60% or less.

[0094] In still further embodiments, the compound of Formula V, Va, or Vb has an average percent maximal efficacy against a drug-sensitive cell line of Table 4 of about 60% or greater.

[0095] In some embodiments, the present invention relates to any compound described herein, wherein C1-C4 alkyl, at each occurrence, is independently methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, tert-butyl, pentyl, or hexyl. In preferred embodiments, C1-C4 alkyl is methyl, ethyl, n-propyl, or iso-propyl.

[0096] In certain embodiments, the present invention relates to any compound described herein, wherein C3-C6 cycloalkyl, at each occurrence, is independently cyclopropyl, cyclobutyl, or cyclopentyl.

[0097] In some embodiments, the present invention relates to any compound described herein, wherein C3-C7 heterocyclyl is, independently at each occurrence, aziridinyl, azetidinyl, diazetidinyl, oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, tetrahydropyranyl, morpholinyl, 2-oxa-6-azaspiro[3.3]heptanyl, 2,6-diazaspiro[3.3]heptanyl, or azaspiro[3.3]heptanyl.

[0098] In some embodiments, the present invention relates to any compound described herein, wherein X1 is CH.

[0099] In some embodiments, the present invention relates to any compound described herein, wherein X2 is CH. Sometimes, X2 is N.

[0100] In certain embodiments, the present invention relates to any of the compounds described herein, wherein V is N.

[0101] In some embodiments, the present invention provides a method for the preparation of b In some cases, R b is NH2.

[0102] In some embodiments, the present invention relates to any compound described herein, wherein Z is CH2.

[0103] In some embodiments, the present invention relates to any compound described herein, wherein Y2 is N(H). Sometimes, Y2 is O.

[0104] In some embodiments, the present invention relates to any compound described herein, wherein R1 is Cl or NH2. Sometimes, R1 is Cl. Sometimes, R1 is NH2.

[0105] In some embodiments, the present invention relates to any compound described herein, wherein R2 is Cl or Br. Sometimes, R2 is Br.

[0106] In some embodiments, the present invention provides a medicament for use in a pharmaceutical composition comprising: b is NH2, Z is CH2, Y2 is N(H), R1 is Cl or NH2, and R2 is Br.

[0107] In some embodiments, the invention relates to compounds having the structure of Formula I, Ia, Ib, II, IIa, IIb, III, IIIa, or IIIb, where m is 0, 1, or 2, and more preferably, m is 0 or 1.

[0108] In a preferred embodiment, the present invention relates to a compound in which R is N(R a )2, and R a One case of is H, and R a is C1-C4 alkyl, preferably methyl.

[0109] In some embodiments, the present invention provides

[0110] [ka] TIFF0007609798000044.tif249165TIFF0007609798000045.tif249160TIFF0007609798000046.tif 230166TIFF0007609798000047.tif211157TIFF0007609798000048.tif213153TIFF000760979800004 9.tif252155TIFF0007609798000050.tif205152TIFF0007609798000051.tif206165TIFF0007609798000052.tif241162TIFF0007609798000053.tif52149, or a pharma- ceutically acceptable salt thereof.

[0111] In some embodiments, the present invention provides

[0112] [ka] TIFF0007609798000055.tif247150TIFF0007609798000056.tif254163TIFF0007609798000057.tif251163TI FF0007609798000058.tif251157TIFF0007609798000059.tif224163TIFF0007609798000060.tif212163TIFF 0007609798000061.tif218157TIFF0007609798000062.tif253155TIFF0007609798000063.tif251162TIFF0007609798000064.tif229161TIFF0007609798000065.tif130155, or a pharma- ceutically acceptable salt thereof.

[0113] In some embodiments, the present invention provides

[0114] [ka] or a pharma- ceutically acceptable salt thereof.

[0115] In some embodiments, the present invention provides

[0116] [ka] TIFF0007609798000069.tif246163TIFF0007609798000070.tif157138, or a pharma- ceutically acceptable salt thereof.

[0117] In some embodiments, the present invention provides

[0118] [ka] The present invention relates to a compound of Formula V having a structure selected from: TIFF0007609798000072.tif102151, or a pharma- ceutically acceptable salt thereof.

[0119] In some embodiments, the present invention provides

[0120] [ka] or a pharma- ceutically acceptable salt thereof.

[0121] In a further aspect, the present invention provides a method for producing a method for the treatment of a cancer

[0122] [ka] The present invention relates to a compound of Formula V having a structure selected from: TIFF0007609798000075.tif102159, or a pharma- ceutically acceptable salt thereof.

[0123] In some embodiments, the present invention provides

[0124] [ka] or a pharma- ceutically acceptable salt thereof.

[0125] In certain embodiments, the compound is

[0126] [ka] TIFF0007609798000085.tif199160 or a pharma- ceutically acceptable salt thereof.

[0127] In other particular embodiments, the compound is

[0128] [ka] or a pharma- ceutically acceptable salt thereof.

[0129] In other particular embodiments, the compound is

[0130] [ka] or a pharma- ceutically acceptable salt thereof.

[0131] In yet another particular embodiment, the compound is

[0132] [ka] or a pharma- ceutically acceptable salt thereof.

[0133] In yet another particular embodiment, the compound is

[0134] [ka] or a pharma- ceutically acceptable salt thereof.

[0135] In yet another particular embodiment, the compound is

[0136] [ka] or a pharma- ceutically acceptable salt thereof.

[0137] In yet another particular embodiment, the compound is

[0138] [ka] or a pharma- ceutically acceptable salt thereof.

[0139] In yet another particular embodiment, the compound is

[0140] [ka] or a pharma- ceutically acceptable salt thereof.

[0141] In yet another particular embodiment, the compound is

[0142] [ka] or a pharma- ceutically acceptable salt thereof.

[0143] In yet another embodiment, the compound is

[0144] [ka] or a pharma- ceutically acceptable salt thereof.

[0145] In certain embodiments, the compound is selected from Table 2.

[0146] In certain embodiments, the present invention relates to a pharmaceutical composition comprising any of the compounds described herein and a pharma- ceutically acceptable diluent or excipient.

[0147] Exemplary Procedures / Methods of Use The compounds described herein are inhibitors of PRMT5 and may therefore be useful for treating diseases whose underlying pathology is mediated (at least in part) by dysregulation of PRMT5 or its normal activity, including cancer and other diseases in which there is a disorder of transcription, cell proliferation, apoptosis, or differentiation.

[0148] In certain embodiments, the method of treating cancer in a subject in need thereof comprises administering to the subject an effective amount of any of the compounds described herein, or a pharma- ceutically acceptable salt thereof.For example, the cancer may be selected from carcinomas (e.g., carcinomas of the endometrium, bladder, breast, colon (e.g., colorectal cancer, e.g., colon adenocarcinoma and colon adenoma), sarcomas (e.g., Kaposi's sarcoma, osteosarcoma, tumors of mesenchymal origin, e.g., fibrosarcoma or rhabdomyosarcoma), kidney, epidermis, liver, lung (e.g., adenocarcinoma, small cell lung carcinoma and non-small cell lung carcinoma), esophagus, gallbladder, ovary, pancreas (e.g., exocrine pancreatic carcinoma), stomach, cervix, thyroid, nose, head and neck, prostate, and skin (e.g., squamous cell carcinoma), human breast cancer (e.g., primary breast tumors, node-negative breast cancer, invasive ductal adenocarcinoma of the breast, non-endometrioid breast cancer), familial melanoma, and melanoma. Other examples of cancers that can be treated with the compounds of the present invention include hematopoietic tumors of lymphoid lineage (e.g., leukemia, acute lymphocytic leukemia, mantle cell lymphoma, chronic lymphocytic leukemia, B cell lymphoma (e.g., diffuse large B cell lymphoma), T cell lymphoma, multiple myeloma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma, and Burkitt's lymphoma), and hematopoietic tumors of myeloid lineage, such as acute and chronic myeloid leukemia, myelodysplastic syndrome, and promyelocytic leukemia. Other cancers include tumors of the central or peripheral nervous system, such as astrocytoma, neuroblastoma, glioma or neurinoma, seminoma, teratocarcinoma, xeroderma pigmentosum, retinoblastoma, keratoacanthoma, and follicular thyroid carcinoma.

[0149] In certain embodiments, the cancer to be treated is selected from colorectal cancer, head and neck cancer, pancreatic cancer, sarcoma, melanoma, myeloma, lymphoma, lung cancer (including non-small cell lung cancer and small cell lung cancer), breast cancer, ovarian cancer, liver cancer, gastric cancer, endometrial cancer, renal cancer, bladder cancer, and acute myeloid leukemia.

[0150] In more specific embodiments, the cancer being treated is selected from colorectal cancer, head and neck cancer, pancreatic cancer, sarcoma, melanoma, myeloma, lymphoma, non-small cell lung cancer, breast cancer, ovarian cancer, liver cancer, gastric cancer, bladder cancer, and acute myeloid leukemia.

[0151] In some embodiments, the subject is a mammal, such as a human.

[0152] Further disclosed herein is a method for inhibiting PRMT5 in a cell, comprising contacting the cell with any of the compounds described herein or its pharma- ceutically acceptable salts, such that PRMT5 enzyme is inhibited in the cell.For example, the cell is a cancer cell.In a preferred embodiment, the proliferation of the cell is inhibited or cell death is induced.

[0153] Further disclosed herein is a method for treating a disease that can be treated by inhibition of PRMT5 in a subject (e.g., a disease characterized by increased expression of PRMT5), comprising administering an effective amount of any of the compounds described herein and / or pharma- ceutically acceptable salts thereof to a subject in need of such treatment. Diseases that can be treated by inhibition of PRMT5 (e.g., a disease characterized by increased expression of PRMT5) include, for example, cancer, metabolic disease, and hematological disease. Further exemplary diseases include colorectal cancer, head and neck cancer, pancreatic cancer, sarcoma, melanoma, myeloma, lymphoma, lung cancer (including non-small cell lung cancer and small cell lung cancer), breast cancer, ovarian cancer, liver cancer, gastric cancer, endometrial cancer, kidney cancer, bladder cancer, and acute myeloid leukemia.

[0154] The method of treatment includes administering a compound of the invention or a pharma- ceutically acceptable salt thereof to a subject in need thereof. Individual embodiments include a method of treating any one of the above disorders or diseases by administering an effective amount of a compound of the invention or a pharma- ceutically acceptable salt thereof to a subject in need thereof.

[0155] Certain embodiments include a method of modulating PRMT5 activity in a subject, comprising administering to the subject a compound of the present invention or a pharma- ceutically acceptable salt thereof. Additional embodiments provide a method for treating a disorder or disease mediated by PRMT5 in a subject in need of such treatment, comprising administering to the subject an effective amount of a compound of formula I, Ia, Ib, II, IIa, IIb, III, IIIa, IIIb, IV, IVa, IVb, V, Va or Vb, or a pharma- ceutically acceptable salt thereof. Other embodiments of the present invention provide a method for treating a disorder or disease mediated by PRMT5 in a subject in need of such treatment, comprising administering to the subject an effective amount of a compound of the present invention or a pharma- ceutically acceptable salt thereof, wherein the disorder or disease is selected from carcinomas with genetic abnormalities that activate PRMT5 activity. These include, but are not limited to, cancer.

[0156] The present invention also provides the use of a compound of the present invention, or a pharma- ceutically acceptable salt thereof, for the treatment of a disorder or disease mediated by PRMT5.

[0157] In some embodiments, the compounds of the invention, or pharma- ceutically acceptable salts thereof, are used to treat disorders or diseases mediated by PRMT5.

[0158] Yet another embodiment of the method provides a compound of Formula I, Ia, Ib, II, IIa, IIb, III, IIIa, IIIb, IV, IVa, IVb, V, Va or Vb, or a pharma- ceutically acceptable salt thereof, for use as a medicament.

[0159] Yet other embodiments of the method include the use of a compound of Formula I, Ia, Ib, II, IIa, IIb, III, IIIa, IIIb, IV, IVa, IVb, V, Va or Vb, or a pharma- ceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of a disorder or disease mediated by PRMT5.

[0160] Certain embodiments of the present invention include the compounds listed in Table 1. Disclosed for each compound in Table 1 is an identifying number ("Compound"), a chemical structure ("Structure"), an exemplary method used to synthesize the compound ("Method"), and the elution order in a chromatographic separation process, as measured, and compared to the diastereomers of that compound ("Elution").

[0161] Certain embodiments of the present invention include compounds of formula IV, wherein V, X1, X2, Y2, Z, R1, R2, and R b is defined as listed in Table 2 for each compound.

[0162] [Table 1] TIFF0007609798000096.tif239161TIFF0007609798000097.tif247163TIFF0007609798 000098.tif249163TIFF0007609798000099.tif249162TIFF0007609798000100.tif53161

[0163] [Table 2] TIFF0007609798000102.tif251152TIFF0007609798000103.tif249140TIFF0007609798000104.tif251141 TIFF0007609798000105.tif250141TIFF0007609798000106.tif252147TIFF0007609798000107.tif253149 TIFF0007609798000108.tif251151TIFF0007609798000109.tif249151TIFF0007609798000110.tif249152 TIFF0007609798000111.tif250150TIFF0007609798000112.tif251146TIFF0007609798000113.tif150157

[0164] Equivalent While specific embodiments of the present invention have been described, the above specification is illustrative and not restrictive. Many variations of the present invention will become apparent to those skilled in the art upon review of this specification and the following claims. The full scope of the present invention should be determined by reference to the claims, along with their full scope of equivalents, and the specification, along with such variations. EXAMPLES

[0165] Example Synthesis Protocol The compounds disclosed herein can be synthesized by several specific methods. The examples outlining specific synthetic routes, and the following general schemes, are intended to provide guidance to a synthetic chemist of ordinary skill, who will readily understand that solvents, concentrations, reagents, protecting groups, order of synthetic steps, times, temperatures, and the like, may be varied as necessary and are well within the skill and judgment of the person skilled in the art.

[0166] [Example 1] Synthesis of compounds 1-10 Preparation of intermediate 1-1

[0167] [ka] The starting material tert-butyl (R)-((1-allyl-2-oxocyclopentyl)methyl)carbamate was prepared according to the procedure of Y. Numajiri, BP Pritchett, K. Chiyoda, and BM Stoltz published in J. Am. Chem. Soc. (2015), Vol. 137, pp. 1040-1043. A solution of tert-butyl (R)-((1-allyl-2-oxocyclopentyl)methyl)carbamate (16.19 g, 64.01 mmol) in tetrahydrofuran (25 mL) was added dropwise via cannula to a cold (-78 °C) solution of lithium hexamethyldisilazide (280 mL, 0.5 M solution in tetrahydrofuran, 140 mmol). After 15 min, a solution of phenylselenium chloride (14.61 g, 76.30 mmol) in tetrahydrofuran (25 mL) was added rapidly via cannula.

[0168] After 30 min at constant temperature, the reaction was quenched by the addition of saturated aqueous ammonium chloride solution (250 mL) and diluted with diethyl ether (500 mL). The mixture was transferred to a separatory funnel. The aqueous phase was separated and the organic phase was further washed with 0.5 N HCl, water, and 3:1 brine / saturated sodium bicarbonate solution (400 mL portions). The organic phase was dried over sodium sulfate, filtered, and concentrated to give an orange oil (35.49 g crude) which was used without further purification.

[0169] The crude product from the previous step was dissolved in dimethylacetamide (200 mL) and eight reactions were carried out sequentially (25 mL each). To each solution behind a blast shield was added pyridine (1.93 mL, 24.0 mmol) followed by (after cooling to 0 °C) 35% aqueous hydrogen peroxide (2.69 mL, 32.0 mmol). The reactions were allowed to warm to room temperature resulting in a strong exotherm, with the reaction temperature increasing from room temperature to 85-90 °C. After the reaction temperature returned to room temperature, each reaction was poured into diethyl ether (150 mL) and saturated sodium sulfite (37.5 mL) and water (37.5 mL) were added. After stirring for 30 minutes, the mixture was transferred to a separatory funnel. The aqueous phase was separated and the organic phase was washed further with water and saturated sodium bicarbonate (75 mL portions). The organic washes from all eight reactions were combined, dried over sodium sulfate, filtered, and concentrated to dryness.

[0170] Silica gel chromatography (10-80% EtOAc / Hexanes) was performed with refractive index detection. The product could also be detected by potassium permanganate staining on TLC. Product fractions were pooled and concentrated to give tert-butyl (S)-((1-allyl-2-oxocyclopent-3-en-1-yl)methyl)carbamate (Intermediate 1-1) as a yellow oil (6.870 g, 43% yield for two steps). 1 H NMR (400 MHz, chloroform-d) δ 7.71 (dt, J = 5.6, 2.7 Hz, 1H), 6.15 (dt, J = 5.8, 2.2 Hz, 1H), 5.60 (dddd, J = 16.8, 10.1, 8.1, 6.7 Hz, 1H), 5.18 - 4.98 (m, 2H), 4.85 (s, 1H), 3.45 - 3.05 (m, 2H), 2.61 (t, J = 2.4 Hz, 2H), 2.44 - 2.12 (m, 2H), 1.41 (s, 9H) ppm. LCMS: [M+H] + m / z = 252.2 amu.

[0171] Preparation of intermediate 1-2

[0172] [ka] To a cold (-78°C) solution of intermediate 1-1 (6.870 g, 27.37 mmol) in methanol (150 mL) was added cerium trichloride heptahydrate (15.30 g, 41.06 mmol). The reaction was stirred for 15 minutes and sodium borohydride was added (1.040 g, 27.37 mmol). The reaction was kept in the cooling bath for 2 hours and then allowed to warm slowly to ambient temperature. When the reaction was complete by TLC analysis, saturated ammonium chloride (200 mL) and water (200 mL) were added. The mixture was transferred to a separatory funnel and the aqueous phase was washed three times with dichloromethane (200 mL portions). The combined organic layers were dried over sodium sulfate, filtered, and concentrated to give a yellow oil.

[0173] Silica gel chromatography (10-30% hexane / acetone) was performed with refractive index detection. The product could also be detected by potassium permanganate staining on TLC. The major desired isomer, intermediate 1-2, eluted first (3.10 g, 45% yield), followed by the minor isomer (2.05 g, 30% yield). The minor isomer could be recycled by pyridinium chlorochromate (PCC) oxidation (2 equivalents of PCC, CHCl, quantitative) and Luche reduction to give additional tert-butyl (((1S,2S)-1-allyl-2-hydroxycyclopent-3-en-1-yl)methyl)carbamate (intermediate 1-2) (1.34 g, 19%, total: 4.44 g, 64% yield). 1 H NMR (400 MHz, chloroform-d) δ 5.97 - 5.68 (m, 3H), 5.20 - 4.90 (m, 2H), 4.33 (s, 1H), 3.45 - 3.09 (m, 2H), 2.30 - 2.00 (m, 4H), 1.44 (s, 9H) ppm. LCMS: [M+H] + m / z = 254.2 amu.

[0174] Preparation of intermediates 1-3

[0175] [ka] A solution of intermediate 1-2 (3.10 g, 12.25 mmol) in dichloromethane (120 mL) was cooled to 0° C. and treated with tert-butyl hydrogen peroxide (5.5 M in decane, 3.34 mL, 18.37 mmol). The reaction was stirred for 5 min and vanadyl acetylacetonate (390 mg, 1.47 mmol) was added as a solid in one portion. The reaction was removed from the cooling bath and stirred at ambient temperature for 8 h, at which point TLC analysis showed complete conversion to the desired product.

[0176] The reaction was concentrated onto silica gel. Silica gel chromatography was performed with refractive index detection (10-50% hexane / acetone). Product fractions were pooled and concentrated to give intermediates 1-3 as yellow oils (2.093 g, 65% yield). 1 H NMR (400 MHz, chloroform-d) δ 5.80 (ddt, J = 16.6, 10.5, 7.3 Hz, 1H), 5.27 (s, 1H), 5.17 - 5.05 (m, 2H), 4.04 (s, 1H), 3.59 (t, J = 2.5 Hz, 1H), 3.47 (t, J = 2.5 Hz, 1H), 3.23 - 3.06 (m, 2H), 2.25 (ddt, J = 13.7, 7.2, 1.2 Hz, 1H), 2.13 - 2.05 (m, 1H), 1.93 (d, J = 15.3 Hz, 1H), 1.80 (dd, J = 15.2, 2.1 Hz, 1H), 1.44 (s, 9H) ppm. LCMS: [M+H] + m / z = 270.2 amu.

[0177] Preparation of intermediates 1-4

[0178] [ka] A round bottom flask was charged with intermediate 1-3 (1.857 g, 6.90 mmol) and ethyl acetate (70 mL) was added to give a clear yellow solution. The reaction was stirred vigorously under nitrogen and sodium azide was added (1.65 g, 20.7 mmol) followed 5 min later by cerium(III) triflate (2.35 g, 4.18 mmol). The mixture was warmed to 50° C. for 2 h. TLC analysis indicated that the epoxide had been consumed.

[0179] The reaction was concentrated and dry loaded onto silica gel. Silica gel chromatography was performed with refractive index detection (10-30% hexanes / acetone). Product fractions were pooled and concentrated to give tert-butyl (((1S,2R,3S,4R)-1-allyl-4-azido-2,3-dihydroxycyclopentyl)methyl)carbamate (Intermediate 1-4) as a clear oil (1.254 g, 58% yield). 1 H NMR (400 MHz, chloroform-d) δ 5.80 (ddt, J = 16.6, 10.5, 7.3 Hz, 1H), 5.27 (s, 1H), 5.19 - 5.05 (m, 2H), 4.04 (s, 1H), 3.59 (t, J = 2.5 Hz, 1H), 3.47 (t, J = 2.4 Hz, 1H), 3.19 (dd, J = 14.2, 7.2 Hz, 1H), 3.13 (dd, J = 14.2, 5.2 Hz, 1H), 2.81 (s, 1H), 2.29 - 2.20 (m, 1H), 2.13 - 2.04 (m, 1H), 1.93 (d, J = 15.2 Hz, 1H), 1.80 (dd, J = 15.2, 2.1 Hz, 1H), 1.44 (s, 9H) ppm. LCMS: [M+H] + m / z = 313.2 amu.

[0180] Preparation of intermediates 1-5

[0181] [ka] A flask containing intermediate 1-4 (1.254 g, 4.005 mmol) was charged with acetone (20 mL) and 2,2'-dimethoxypropane (20 mL), followed by p-toluenesulfonic acid monohydrate (38.1 mg, 0.20 mmol). The reaction was stirred at room temperature for 2 h and quenched by the addition of pyridine (0.020 mL).

[0182] Silica gel was added to the solution and the volatiles were removed in vacuo. The residue was dry loaded and purified by silica gel chromatography with refractive index detection (0-15% hexanes / diethyl ether). Product fractions were pooled and concentrated to give a clear oil (1.253 g, 89% yield).

[0183] The clear oil product of the previous step (1.253 g, 3.53 mmol) was transferred to a round bottom flask and dissolved in tetrahydrofuran (9 mL) under nitrogen atmosphere. Water (0.64 mL, 35.3 mmol) was injected followed by trimethylphosphine (8.8 mL, 1 M solution in tetrahydrofuran, 8.8 mmol). The reaction was stirred for 2 h and the volatiles were removed first under a stream of nitrogen and then in vacuo. To the resulting crude amine was added a mixture of Hunig's base (3.08 mL, 17.65 mmol) and iso-propyl alcohol (15 mL) under nitrogen atmosphere. To the resulting solution was added 2-(4,6-dichloropyrimidin-5-yl)acetaldehyde (1.011 g, 5.295 mmol). The reaction was sealed and warmed to 75° C. for 20 h. At this point the reaction was complete by TLC analysis and was diluted with ethyl acetate (150 mL) and transferred to a separatory funnel The organic phase was washed successively with water and saturated aqueous ammonium chloride (50 mL portions), dried over sodium sulfate, filtered and concentrated onto silica gel.

[0184] The residue was dry loaded and purified by silica gel chromatography with refractive index detection (0-100% hexanes / EtOAc). Product fractions were pooled and concentrated to give tert-butyl (((3aR,4S,6R,6aS)-4-allyl-6-(4-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-4H-cyclopenta[d][1,3]dioxol-4-yl)methyl)carbamate (Intermediate 1-5) as a clear oil (841 mg, 52% yield). 1 H NMR (400 MHz, chloroform-d) δ 8.64 (s, 1H), 7.30 (d, J = 3.7 Hz, 1H), 6.62 (d, J = 3.7 Hz, 1H), 6.08 - 5.76 (m, 1H), 5.22 - 5.02 (m, 4H), 4.57 (d, J = 6.3 Hz, 1H), 3.53 (dd, J = 14.0, 8.6 Hz, 1H), 3.29 (dd, J = 14.1, 4.4 Hz, 1H), 2.51 - 2.33 (m, 2H), 2.27 (dd, J = 14.0, 8.0 Hz, 1H), 2.17 (dd, J = 13.8, 7.1 Hz, 1H), 1.61 (s, 3H), 1.46 (s, 9H), 1.32 (s, 3H) ppm. LCMS: [M+H] + m / z = 463.2 amu.

[0185] Preparation of Intermediate 1-6 R&S

[0186] [ka] A flask containing intermediate 1-5 (841 mg, 1.81 mmol) was charged with tert-butanol and water to give a biphasic solution (1:1, 20 mL). AD-mix-β was added (2.54 g, 1.4 g / mmol of substrate), followed by hydroquinidine, 1,4-phthalazinediyl diether (DHQD2PHAL) (42.3 mg, 0.0543 mmol) and osmium tetroxide (0.345 mL, 4% w / w aqueous solution, 0.0543 mmol). The reaction was stirred for 4 h and poured into a separatory funnel containing 1N aqueous sodium sulfite solution (50 mL) and ethyl acetate (100 mL). The aqueous phase was separated and the organic phase was further washed with 0.2 N HCl, 0.2 N NaOH and brine (50 mL portions). The organic phase was concentrated in vacuo and used without further purification.

[0187] The resulting crude diol was dissolved in tert-butanol (30 mL) and pH 7 phosphate buffer (20 mL). Sodium periodate (675 mg, 1.81 mmol) was added in one portion and the reaction was complete within 30 min as determined by TLC analysis. The reaction was poured into a separatory funnel containing ethyl acetate (100 mL) and saturated sodium bicarbonate (50 mL). The organic layer was dried over sodium sulfate, filtered and concentrated.

[0188] To the resulting oil was added methanol (20 mL) and pyridinium p-toluenesulfonate (PPTS) (45 mg, 0.18 mmol). The reaction was stirred for 3 h and TLC showed the starting material was consumed. Silica gel was added to the solution and the volatiles were removed in vacuo. The residue was dry loaded and purified by silica gel chromatography with refractive index detection (0-50% hexanes / EtOAc). Product fractions were pooled and concentrated to give intermediates 1-6 R&S as a mixture of stereoisomeric yellow oils (345 mg, 40% yield over three steps). 1H NMR (400 MHz, Chloroform-d) δ 8.63 - 8.53 (m, 1H), 7.28 (m, 1H), 6.57 (m, 1H), 5.52 - 4.93 (m, 2H), 4.93 - 4.38 (m, 1H), 4.03 - 3.59 (m, 2H), 3.42 (s, 3H), 3.36 - 3.02 (m, 1H), 2.75 - 2.38 (m, 2H), 2.40 - 1.80 (m, 2H), 1.44 (m, 9H), 1.24 - 1.19 (m, 6H) ppm. LCMS: [M+H] + m / z = 479.2 amu.

[0189] Preparation of Compounds 1 and 2: Step 1

[0190] [ka] A flame-dried vial was charged with 3-bromo-2-chloro-7-iodoquinoline (129.2 mg, 0.351 mmol) and charged with dry tetrahydrofuran (1 mL) under nitrogen atmosphere. After cooling (-78 °C), a solution of iso-propylmagnesium chloride (0.18 mL, 2 M in tetrahydrofuran, 0.36 mmol) was charged. After 10 min, the resulting solution was transferred via syringe to a cold (-78 °C) slurry of copper(I) bromide dimethylsulfide in tetrahydrofuran (1 mL). After 15 min, boron trifluoride diethyl etherate was charged (43 μL, 0.35 mmol) and after another 15 min, a solution of intermediate 1-6 R&S (42 mg, 0.087 mmol) in diethyl ether (6 mL) was charged.

[0191] The reaction was allowed to warm gradually over 2 h, at which point LC / MS analysis indicated consumption of starting material. The reaction was quenched with saturated aqueous ammonium chloride and ammonium hydroxide (2 mL portions). The resulting mixture was poured into a separatory funnel containing ethyl acetate (50 mL), and the organic phase was washed with water and brine (50 mL portions). The organic phase was concentrated onto silica gel, and the residue was purified by chromatography (5-20% hexanes / acetone). Two fractions were collected, the minor diastereomer eluting first (12 mg, 15% yield), followed by the major diastereomer eluting first (36 mg, 45% yield). Intermediate 1-7 R&S Fraction 1 (minor diastereomer): 1 H NMR (400 MHz, chloroform-d) δ 8.61 (d, J = 7.8 Hz, 1H), 8.42 (s, 1H), 7.83 (s, 1H), 7.74 (d, J = 8.5 Hz, 1H), 7.52 - 7.46 (m, 2H), 7.29 (d, J = 3.6 Hz, 1H), 6.62 (d, J = 3.6 Hz, 1H), 5.21 (t, J = 6.1 Hz, 1H), 4.91 (dt, J = 11.7, 6.6 Hz, 1H), 4.82 (t, J = 8.5 Hz, 1H), 4.71 (d, J = 7.1 Hz, 1H), 3.96 (d, J = 11.6 Hz, 1H), 3.79 (d, J = 11.5 Hz, 1H), 2.68 (t, J = 12.1 Hz, 1H), 2.47 (m, 2H), 1.27 (m, 6H), 1.06 (s, 9H) ppm. LCMS: [M+H] + m / z = 688.1 amu. Intermediate 1-7 R&S Fraction 2 (Major Diastereomer): 1H NMR (400 MHz, chloroform-d) δ 8.53 (s, 1H), 8.31 (d, J = 14.2 Hz, 1H), 7.80 (d, J = 4.8 Hz, 1H), 7.63 (t, J = 7.8 Hz, 1H), 7.41 (dd, J = 13.7, 8.1 Hz, 1H), 7.20 (s, 1H), 6.52 (d, J = 3.6 Hz, 1H), 5.18 - 4.70 (m, 3H), 4.59 (dd, J = 16.7, 7.4 Hz, 1H), 4.35 - 4.17 (m, 1H), 3.33 (dd, J = 10.8, 3.0 Hz, 1H), 2.71 - 1.82 (m, 2H). 1.40 (s, 3H), 1.27 (s, 3H), 1.05 (s, 9H) ppm. LCMS: [M+H] + m / z = 688.1 amu.

[0192] Preparation of Compounds 1 and 2: Step 2

[0193] [ka] Both diastereomers prepared in step 1 were subjected to the following general procedure. The vial containing the first fraction of intermediate 1-7 R&S (10.6 mg, 0.015 mmol) was dissolved in methanol (1 mL) to give a clear solution, followed by treatment with 4N hydrochloric acid in dioxane (50 μL, 0.15 mmol). The reaction was stirred at room temperature for 90 min, concentrated to dryness, and the residue was dissolved in DMSO.

[0194] After filtration, reverse phase HPLC was performed (5-50% acetonitrile / water + 0.25% acetic acid, 20 mm x 250 mm C18 column, 40 mL / min gradient over 15 min, detection wavelength of 242 nM, total of 3 injections). Product fractions were pooled, frozen and concentrated on a lyophilizer to give the product as a white solid (3.0 mg, 38% yield). Fraction 1 (compound 2): 1H NMR (400 MHz, アセトニトリル-d3) δ 8.52 (s, 1H), 8.50 (s, 1H), 7.86 (dd, J = 1.7, 0.9 Hz, 1H), 7.77 (d, J = 8.5 Hz, 1H), 7.66 (dd, J = 8.5, 1.7 Hz, 1H), 7.55 - 7.44 (m, 1H), 6.59 (d, J = 3.7 Hz, 1H), 5.11 - 4.91 (m, 1H), 4.49 - 4.32 (m, 2H), 3.83 (d, J = 4.7 Hz, 1H), 3.46 (d, J = 10.6 Hz, 1H), 2.80 (d, J = 10.6 Hz, 1H), 2.48 - 2.33 (m, 2H), 1.75 - 1.64 (m, 2H) ppm. LCMS: [M+H] + m / z = 548.2 amu. Drawing points 2 (compound 1): 1 H NMR (400 MHz, アセトニトリル-d3) δ 8.61 (s, 1H), 8.49 (s, 1H), 8.01 (s, 1H), 7.91 (d, J = 8.5 Hz, 1H), 7.67 (dd, J = 8.5, 1.9 Hz, 1H), 7.50 (d, J = 4.2 Hz, 1H), 6.58 (d, J = 3.7 Hz, 1H), 4.91 (q, J = 9.3 Hz, 1H), 4.55 (dd, J = 8.7, 5.0 Hz, 1H), 4.07 - 3.95 (m, 2H), 3.46 - 3.21 (m, 1H), 2.70 (dd, J = 13.6, 6.4 Hz, 1H), 2.55 - 2.38 (m, 2H), 2.05 - 2.00 (m, 2H) ppm. LCMS: [M+H] + m / z = 548.2 amu.

[0195] Preparation of compounds 3 and 4

[0196]

change

[0197] The residue was taken up in benzene (2 mL) and concentrated again. To the crude oil was added methanol (0.9 mL) and 4N hydrochloric acid in dioxane (0.1 mL). After 45 min, the yellow solution was concentrated, dissolved in DMSO (1 mL) and purified by reverse phase HPLC (5-25% acetonitrile / water + 0.25% acetic acid, 20 mm x 250 mm C18 column, 40 mL / min gradient over 15 min, detection wavelength of 242 nM, total of 5 injections). Product fractions were pooled, frozen and concentrated on a lyophilizer to give the product as a white solid (1.1 mg, 37% yield). Fraction 1 (compound 4): 1 H NMR (400 MHz, acetonitrile-d3) δ 8.51 (s, 1H), 8.06 (s, 1H), 7.85 (dd, J = 1.7, 0.8 Hz, 1H), 7.77 (d, J = 8.5 Hz, 1H), 7.65 (dd, J = 8.5, 1.6 Hz, 1H), 7.10 (d, J = 3.6 Hz, 1H), 6.41 (d, J = 3.6 Hz, 1H), 5.62 (s, 2H), 4.96 - 4.75 (m, 1H), 4.45 - 4.35 (m, 1H), 4.31 (dd, J = 7.7, 4.8 Hz, 1H), 3.81 (d, J = 4.8 Hz, 1H), 3.45 (d, J = 10.6 Hz, 1H), 2.79 (d, J = 10.7 Hz, 1H), 2.36 (ddd, J = 13.7, 8.6, 4.8 Hz, 2H), 1.74 - 1.61 (m, 2H) ppm. LCMS: [M+H] + m / z = 529.1 amu. Fraction 2 (compound 3):1 H NMR (400 MHz, アセトニトリル-d3) δ 8.51 (s, 1H), 8.02 (s, 1H), 7.87 (d, J = 1.5 Hz, 1H), 7.77 (d, J = 8.5 Hz, 1H), 7.65 (dd, J = 8.6, 1.7 Hz, 1H), 6.99 (d, J = 3.6 Hz, 1H), 6.36 (d, J = 3.6 Hz, 1H), 5.61 (s, 2H), 4.75 (q, J = 9.0 Hz, 1H), 4.53 (t, J = 8.3 Hz, 1H), 4.37 (dd, J = 8.0, 5.1 Hz, 1H), 3.88 (d, J = 4.9 Hz, 1H), 3.53 (d, J = 10.9 Hz, 1H), 2.86 (d, J = 10.8 Hz, 1H), 2.44 (dd, J = 12.8, 7.5 Hz, 1H), 2.32 (dd, J = 13.6, 9.6 Hz, 1H), 2.05 - 2.00 (m, 2H) ppm. LCMS: [M+H] + m / z = 529.1 amu.

[0198] Preparation of compound 5 and 6: ステップ1

[0199]

change

[0200] The reaction was maintained at -45°C for 2 h and then warmed to 0°C over an additional 2 h, at which point LC / MS analysis indicated consumption of starting material. The reaction was quenched with saturated aqueous ammonium chloride and ammonium hydroxide (10 mL portions). The resulting mixture was poured into a separatory funnel containing ethyl acetate (150 mL), and the organic phase was washed with water and brine (50 mL portions). The organic phase was concentrated onto silica gel, and the residue was purified by chromatography (15-45% hexanes / EtOAc). Two fractions were collected, the minor diastereomer eluting first (67.2 mg, 25% yield), followed by the major diastereomer eluting first (158.1 mg, 59% yield). Intermediate 1-8 R&S Fraction 1 - Contains Rotamers (Minor Diastereomers): 1H NMR (500 MHz, クロロホルム-d) δ 8.62 (s, 1H), 8.29 (s, 1H), 7.84 (m, 1H), 7.67 (d, J = 8.5 Hz, 1H), 7.45 (t, J = 9.3 Hz, 1H), 7.29 (d, J = 3.6 Hz, 1H), 7.23 - 7.18 (m, 2H), 6.72 (d, J = 8.5 Hz, 2H), 6.63 (t, J = 7.9 Hz, 1H), 5.30 - 4.87 (m, 4H), 4.86 - 4.67 (m, 1H), 3.98 (d, J = 11.6 Hz, 1H), 3.74 - 3.72 (m, 4H), 2.77 - 2.20 (m, 4H), 1.65 - 1.50 (m, 6H), 1.27 (m, 9H), 1.06 (s, 6H), 0.95 - 0.78 (m, 2H) ppm. LCMS: [M+H] + m / z = 923.2 amu. Intermediate body 1-8 R&S drawing points 2 - Return to the opposite sex (main body): 1 H NMR (500 MHz, クロロホルム-d) δ 8.65 (s, 1H), 8.32 (d, J = 10.7 Hz, 1H), 7.91 (s, 1H), 7.70 (d, J = 8.5 Hz, 1H), 7.48 (s, 1H), 7.29 (m, 8H), 6.78 (d, J = 8.7 Hz, 2H), 6.64 (d, J = 3.7 Hz, 1H), 5.33 - 4.85 (m, 5H), 4.71 (dd, J = 22.2, 7.4 Hz, 1H), 4.49 - 4.30 (m, 1H), 3.79 (s, 3H), 3.57 - 3.36 (m, 1H), 2.82 - 2.46 (m, 1H), 2.44 - 2.30 (m, 1H), 2.06 - 1.97 (m, 2H), 1.69 (m, 3H), 1.54 - 1.41 (m, 9H), 1.18 (s, 3H) ppm. LCMS: [M+H] + m / z = 923.2 amu.

[0201] Preparation of Compounds 5 and 6: Step 2

[0202] [ka] Both diastereomers prepared in step 1 above were subjected to the following general procedure. The vial containing the major diastereomer of intermediate 1-8 R&S (8.0 mg, 0.0087 mmol) was dissolved in dichloromethane (2 mL) and trifluoroacetic acid was injected (1 mL). The reaction was stirred for 90 minutes and poured into a separatory funnel containing saturated sodium bicarbonate. The organic phase was separated and the aqueous phase was washed two more times with dichloromethane. The combined organic extracts were concentrated to dryness and dissolved in chloroform and methanol (1:1, 2 mL). Triacetoxyborohydride (7.0 mg, 0.03 mmol) and 37% formaldehyde (20 μL, 0.021 mmol) were added and the reaction was stirred for 60 minutes. The solution was poured into a separatory funnel containing 1N sodium hydroxide (10 mL) and the aqueous phase was washed three times with dichloromethane.

[0203] After concentration, the crude material was dissolved in dioxane (300 μL) and treated with ammonium hydroxide (300 μL). The reaction vial was sealed and heated to 120° C. for 40 h. The solvent was removed under a stream of nitrogen and then in vacuo. The crude solid was dissolved in trifluoroacetic acid (500 μL) and heated to 100° C. for 40 min. After concentration, the resulting oily solid was dissolved in DMSO and purified by reverse phase HPLC (2.5-25% acetonitrile / water + 0.25% acetic acid, 20 mm×250 mm C18 column, 40 mL / min gradient over 15 min, detection wavelength of 242 nM, total of 3 injections). Product fractions were pooled, frozen and concentrated on a lyophilizer to give the product as a white solid (2.5 mg, 55% yield). Fraction 1 (compound 5): 1H NMR (400 MHz, アセトニトリル-d3) δ 8.25 (s, 1H), 8.12 (s, 1H), 7.62 (d, J = 8.3 Hz, 1H), 7.53 (t, J = 1.1 Hz, 1H), 7.35 (dd, J = 8.3, 1.6 Hz, 1H), 7.14 (d, J = 3.7 Hz, 1H), 6.47 (d, J = 3.6 Hz, 1H), 5.71 (s, 2H), 5.60 (s, 2H), 4.88 (dt, J = 10.0, 7.7 Hz, 1H), 4.35 (dd, J = 7.3, 4.8 Hz, 1H), 3.94 LCMS: [M+H] + m / z = 524.1 amu. Drawing points 2 (compound 6): 1 H NMR (400 MHz, アセトニトリル-d3) δ 8.29 - 8.17 (m, 1H), 8.09 (s, 1H), 7.58 (d, J = 8.3 Hz, 1H), 7.49 (dd, J = 1.6, 0.7 Hz, 1H), 7.32 (dd, J = 8.3, 1.7 Hz, 1H), 7.08 (d, J = 3.6 Hz, 1H), 6.44 (d, J = 3.6 Hz, 1H), 5.71 (s, 2H), 5.58 (s, 2H), 4.92 (dt, J = 10.0, 8.4 Hz, 1H), 4.34 (dd, J = 7.4, 4.8 Hz, 1H), 3.91 (d, J = 4.8 Hz, 1H), 3.36 (t, J = 8.4 Hz, 1H), 3.03 (d, J = 9.7 Hz, 1H), 2.77 (d, J = 9.7 Hz, 1H), 2.67 (dd, J = 13.5, 10.0 Hz, 1H), 2.43 - 2.36 (m, 2H), 2.15 (s, 3H) ppm. LCMS: [M+H] +m / z = 524.1 amu.

[0204] Preparation of Compounds 7 and 8

[0205] [ka] Both diastereomers prepared in step 1 of the preparation of compounds 5 and 6 were subjected to the following general procedure. Intermediate 1-8 The vial containing fraction 1 of R&S was dissolved in dioxane (630 μL) and injected with ammonium hydroxide (630 μL). The reaction vial was sealed and heated to 100° C. for 27 h. The solvent was removed under a stream of nitrogen and then in vacuo. The crude solid was dissolved in trifluoroacetic acid (910 μL), treated with thioanisole (36 μL, 0.30 mmol) and heated to 70° C. for 60 min.

[0206] After concentration in vacuo, the resulting oily solid was dissolved in methanol (2 mL) and treated with potassium carbonate (20.8 mg, 0.15 mmol). The mixture was stirred for 1 h, filtered, concentrated, dissolved in DMSO, and purified by reverse-phase HPLC (3-15% acetonitrile / water + 0.25% acetic acid, 20 mm x 250 mm C18 column, 25 mL / min gradient over 15 min, detection wavelength of 242 nM, total of 5 injections). Product fractions were pooled, frozen and concentrated on a lyophilizer to give the product as a white solid (10.5 mg, 60% yield for 3 steps). Fraction 1 (compound 7): 1H NMR (400 MHz, アセトニトリル-d3) δ 8.23 ​​(s, 1H), 8.14 (s, 1H), 7.71 - 7.49 (m, 2H), 7.34 (dd, J = 8.4, 1.7 Hz, 1H), 7.19 (d, J = 3.7 Hz, 1H), 6.49 (d, J = 3.6 Hz, 1H), 5.71 (s, 2H), 5.57 (s, 2H), 4.92 (dt, J = 9.7, 8.3 Hz, 1H), 4.44 - 4.30 (m, 2H), 3.91 (d, J = 4.9 Hz, 1H), 3.56 (d, J = 10.6 Hz, 1H), 2.83 (d, J = 10.7 Hz, 1H), 2.50 - 2.35 (m, 2H), 1.84 - 1.70 (m, 2H). ppm. LCMS: [M+H] + m / z = 510.1 amu. Drawing points 2 (compound 8): 1 H NMR (400 MHz, アセトニトリル-d3) δ 8.22 (s, 1H), 8.11 (s, 1H), 7.64 - 7.50 (m, 2H), 7.33 (dd, J = 8.3, 1.7 Hz, 1H), 7.08 (d, J = 3.6 Hz, 1H), 6.44 (d, J = 3.6 Hz, 1H), 5.69 (s, 2H), 5.56 (s, 2H), 4.85 (q, J = 8.9 Hz, 1H), 4.43 (dd, J = 8.0, 4.9 Hz, 2H), 3.93 (d, J = 5.0 Hz, 1H), 3.53 (d, J = 10.7 Hz, 1H), 2.90 (d, J = 10.7 Hz, 1H), 2.53 - 2.34 (m, 2H), 2.15 - 2.08 (m, 2H) ppm. LCMS: [M+H] + m / z = 510.1 amu.

[0207] To a suspension of compound 7 (10.5 mg, 0.021 mmol) in water (4 mL) was added a 0.1 N solution of methanesulfonic acid (21 μL, 1.02 equiv, 0.021 mmol). The mixture was sonicated for 60 min, became clear and homogeneous, and was frozen and concentrated on a lyophilizer to give the salt as a white fluffy powder (11.5 mg). 1 H NMR (400 MHz, heavy water) δ 8.51 (s, 1H), 8.15 (s, 1H), 7.79 (d, J = 8.4 Hz, 1H), 7.61 (d, J = 1.6 Hz, 1H), 7.50 - 7.36 (m, 2H), 6.78 (d, J = 3.7 Hz, 1H), 5.11 (q, J = 8.9 Hz, 1H), 4.94 (dd, J = 12.2, 6.3 Hz, 1H), 4.63 - 4.56 (m, 1H), 4.18 (d, J = 4.7 Hz, 1H), 3.86 (d, J = 12.5 Hz, 1H), 3.36 (d, J = 12.5 Hz, 1H), 2.78 - 2.71 (m, 1H), 2.60 (dd, J = 14.2, 10.3 Hz, 1H), 2.56 - 2.46 (m, 1H), 2.22 (dd, J = 14.2, 8.5 Hz, 1H), 1.94 (s, 3H) ppm. LCMS: [M+H] + m / z = 510.1 amu.

[0208] Preparation of Compounds 9 and 10

[0209] [ka] Both diastereomers prepared in step 1 for the preparation of compounds 5 and 6 were subjected to the following general procedure: A vial containing fraction 1 of intermediate 1-8 R&S (67.2 mg, 0.072 mmol) was dissolved in trifluoroacetic acid (3.0 mL), treated with thioanisole (86 μL, 0.72 mmol) and heated to 70° C. for 60 min.

[0210] After concentration in vacuo, the resulting oily solid was dissolved in methanol (3 mL) and treated with potassium carbonate (49.8 mg, 0.36 mmol). The mixture was stirred for 1 h, filtered, concentrated, dissolved in DMSO, and purified by reverse-phase HPLC (5-25% acetonitrile / water + 0.25% acetic acid, 20 mm x 250 mm C18 column, 25 mL / min gradient over 15 min, detection wavelength of 242 nM, total of 5 injections). Product fractions were pooled, frozen and concentrated on a lyophilizer to give the product as a white solid (19.5 mg, 51% yield for two steps). Fraction 1 (Compound 9): 1 H NMR (400 MHz, acetonitrile-d3) δ 8.58 (s, 1H), 8.23 ​​(s, 1H), 7.65 - 7.51 (m, 3H), 7.35 (dd, J = 8.4, 1.6 Hz, 1H), 6.67 (d, J = 3.7 Hz, 1H), 5.57 (s, 2H), 5.19 - 5.03 (m, 1H), 4.49 (dd, J = 7.9, 4.8 Hz, 1H), 4.43 - 4.28 (m, 1H), 3.92 (d, J = 4.7 Hz, 1H), 3.57 (d, J = 10.6 Hz, 1H), 2.84 (d, J = 10.6 Hz, 1H), 2.55 - 2.37 (m, 2H), 1.85 - 1.75 (m, 2H) ppm. LCMS: [M+H] + m / z = 529.1 amu. Fraction 2 (Compound 10): 1H NMR (400 MHz, acetonitrile-d3) δ 8.54 (s, 1H), 8.22 (d, J = 0.7 Hz, 1H), 7.65 - 7.44 (m, 3H), 7.33 (dd, J = 8.3, 1.7 Hz, 1H), 6.62 (d, J = 3.7 Hz, 1H), 5.58 (s, 2H), 5.05 (dt, J = 10.0, 8.8 Hz, 1H), 4.54 (dd, J = 8.5, 4.7 Hz, 1H), 4.51 - 4.46 (m, 1H), 3.95 (d, J = 4.7 Hz, 1H), 3.56 (d, J = 10.7 Hz, 1H), 2.92 (d, J = 10.7 Hz, 1H), 2.54 - 2.42 (m, 2H), 2.14 - 1.99 (m, 2H) ppm. LCMS: [M+H] + m / z = 529.1 amu.

[0211] To a suspension of compound 9 (9.5 mg, 0.017 mmol) in water (4 mL) was added a 0.1 N solution of methanesulfonic acid (18 μL, 1.02 equiv, 0.018 mmol). The mixture was sonicated for 60 min, became clear and homogeneous, and was frozen and concentrated on a lyophilizer to give the salt as a white fluffy powder (10.5 mg). 1H NMR (400 MHz, heavy water) δ 8.75 (s, 1H), 8.57 (s, 1H), 7.95 (d, J = 8.4 Hz, 1H), 7.80 - 7.53 (m, 3H), 6.83 (d, J = 3.7 Hz, 1H), 5.23 (q, J = 8.8 Hz, 1H), 5.05 (dd, J = 12.2, 6.3 Hz, 1H), 4.92 - 4.86 (m, 1H), 4.74 (s, 3H), 4.29 (d, J = 4.7 Hz, 1H), 3.96 (d, J = 12.5 Hz, 1H), 3.47 (d, J = 12.5 Hz, 1H), 2.87 (dd, J = 13.4, 6.3 Hz, 1H), 2.70 (dd, J = 14.2, 10.3 Hz, 1H), 2.65 - 2.55 (m, 1H), 2.33 (dd, J = 14.2, 8.5 Hz, 1H) ppm. LCMS: [M+H] + m / z = 529.1 amu.

[0212] [Example 2] Synthesis of compounds 11~26 Modulation of intermediate 2-1

[0213]

change

[0214] Preparation of intermediate 2-2

[0215] [ka] To an oven-dried flask under argon was added tris(dibenzylideneacetone)dipalladium(0) (575 mg, 0.630 mmol) and (R)-p-(CF3)3-t-BuPHOX (1.11 g, 1.88 mmol). Sequential vacuum-argon cycles (3x) were performed and degassed toluene (450 mL) was injected to give a dark catalyst solution which was stirred for 45 min. Raw intermediate 2-1 (13.15 g, 41.31 mmol) was injected dropwise and the reaction was stirred under argon for 17 h. TLC analysis showed complete conversion and the reaction was filtered through a celite pad and concentrated to give an oil. The oil was dissolved in DCM / Hexane (1:1) and purified by silica gel chromatography, exchanging DCM / Hexane (1:1) to DCM and finally to 5% EtOAc / DCM. Product fractions were pooled and concentrated to dryness to give (R)-2-allyl-2-(((4-methoxybenzyl)oxy)methyl)cyclopentan-1-one (Intermediate 2-2, 10.45 g, 92% yield) as a clear yellow oil. Further analysis by chiral SFC measurement (IC-Daicell, 100 bar, 40° C., 5% IPA / CO2 over 12 min) showed Intermediate 2-2 to be in 80% ee, with peak integration confirming the racemic sample at 210 nM. Proton NMR and LC / MS were consistent with the expected product. 1 H NMR (400 MHz, CDCl3): δ 7.22 - 7.16 (m, 2H), 6.89 - 6.83 (m, 2H), 5.77 - 5.52 (m, 1H), 5.12 - 4.98 (m, 2H), 4.47 - 4.30 (m, 2H), 3.80 LCMS: [M+Na] + m / z = 297.2 amu.

[0216] Preparation of intermediate 2-3

[0217] [ka] A solution of intermediate 2-2 (1.89 g, 1.0 equiv, 6.89 mmol) in dry tetrahydrofuran was treated dropwise via cannula with a cold (-78 °C) solution of lithium hexamethyldisilazide (8.6 mL, 1.25 equiv, 8.6 mmol). After 15 min, a solution of sulfuryl-tert-butylimine chloride (1.78 g, 1.20 equiv, 8.26 mmol) in tetrahydrofuran was added quickly via cannula.

[0218] After 30 min at constant temperature, the reaction was quenched with saturated aqueous ammonium chloride and diluted with diethyl ether. The mixture was transferred to a separatory funnel, the aqueous phase was separated, and the organic phase was further washed with 0.5N HCl, water, and 3:1 brine / saturated sodium bicarbonate solution. The organic phase was dried over sodium sulfate, filtered, and concentrated to give a crude oily product, which was then purified by silica gel chromatography (0-100% EtOAc / hexanes with refractive index detection) to give (S)-5-allyl-5-(((4-methoxybenzyl)oxy)methyl)cyclopent-2-en-1-one (Intermediate 2-3). Intermediate 2-3 also stains with potassium permanganate on TLC. Proton NMR and LC / MS were consistent with the expected product. 1H NMR (400 MHz, CDCl3): δ 7.67 (dt, J = 5.6, 2.7 Hz, 1H), 7.23 - 7.12 (m, 2H), 6.93 - 6.78 (m, 2H), 6.16 (dt, J = 5.8, 2.2 Hz, 1H), 5.56 (dddd, J = 16.7, 10.0, 8.2, 6.5 Hz, 1H), 5.12 - 4.93 (m, 2H), 4.47 - 4.27 (m, 2H), 3.79 (s, 3H), 3.50 (d, J = 8.7 Hz, 1H), 3.36 (d, J = 8.7 Hz, 1H), 2.81 (dt, J = 19.2, 2.5 Hz, 1H), 2.54 (ddd, J = 19.2, 2.8, 2.1 Hz, 1H), 2.26 (ddt, J = 13.6, 6.5, 1.3 Hz, 1H), 2.18 (ddt, J = 13.5, 8.2, 1.0 Hz, 1H)ppm. LCMS: [M+Na] + m / z = 295.2 amu.

[0219] Modulation of intermediate 2-4

[0220]

change

[0221] Preparation of intermediate 2-5

[0222] [ka] Dichloromethane (180 mL, 0.1 M) was added to a flask containing intermediate 2-4 (4.875 g, 1.00 equiv, 17.76 mmol) and cooled to 0° C., followed by the addition of m-CPBA (75% w / w, 6.13 g, 1.50 equiv, 26.65 mmol) and sodium bicarbonate (3.73 g, 2.50 equiv, 44.4 mmol). After 2 h, TLC indicated the reaction was not complete, so the reaction was transferred to a circulating cold bath at −15° C. and stirred overnight (14 h). After stirring overnight, TLC confirmed the reaction was complete. The mixture was poured into sodium sulfite (250 mL, saturated) and stirred for 15 min before being transferred to a separatory funnel. The mixture was diluted with diethyl ether (500 mL) and the aqueous layer was separated. The ether layer was further washed with half-saturated and fully saturated sodium bicarbonate (200 mL portions). The organic phase was dried over magnesium sulfate, filtered and concentrated to dryness to give (1R,2R,3S,5S)-3-allyl-3-(((4-methoxybenzyl)oxy)methyl)-6-oxabicyclo[3.1.0]hexan-2-ol (Intermediate 2-5, 5.50 g, crude, quantitative) as a yellow oil which was pure by LC / MS and proton NMR analysis. 1 H NMR (400 MHz, CDCl3): δ 7.25 - 7.19 (m, 2H), 6.90 - 6.84 (m, 2H), 5.70 (ddt, J = 16.8, 10.3, 7.4 Hz, 1H), 5.12 - 5.05 (m, 2H), 4.47 (d, J = 11.3 Hz, 1H), 4.34 (d, J = 11.4 Hz, 1H), 4.07 (s, 1H), 3.84 - 3.78 (m, 5H), 3.61 (dd, J = 2.9, 1.8 Hz, 1H), 3.42 (dd, J = 2.9, 1.8 Hz, 1H), 3.25 (d, J = 8.9 Hz, 1H), 2.53 - 2.41 (m, 1H), 2.12 (ddt, J = 13.6, 7.2, 1.2 Hz, 1H), 1.82 (d, J = 15.1 Hz, 1H), 1.69 (dd, J = 15.1, 1.9 Hz, 1H) ppm. LCMS: [M+H] + m / z = 291.2 amu.

[0223] Preparation of intermediate 2-6

[0224] [ka] A round bottom flask was charged with intermediate 2-5 (139.4 mg, 1.0 equiv, 0.48 mmol) and dry acetonitrile (0.1 M) to give a clear yellow solution. The reaction was stirred vigorously under nitrogen gas and sodium azide was added (150 mg, 5.0 equiv, 2.3 mmol). After 5 min, cerium(III) triflate (305 mg, 1.1 equiv, 0.52 mmol) was added. The reaction was warmed to 50° C. for a period of 5 h until NMR analysis showed that the epoxide was consumed. The reaction was poured into 30 mL of ethyl acetate and 50% concentrated ammonium chloride (1:1). The mixture was filtered through Celite and the filter cake was washed with ethyl acetate and water. The biphasic mixture was separated and the organic phase was dried over sodium sulfate. The drying agent was filtered off, and the filtrate was concentrated onto silica gel and purified by flash chromatography (10-60% ethyl acetate / hexanes) to give (1S,2R,3S,5R)-3-allyl-5-azido-3-(((4-methoxybenzyl)oxy)methyl)cyclopentane-1,2-diol (intermediate 2-6, 70.0 mg, 45% yield), as confirmed by NMR and LC / MS analyses. 1H NMR (400 MHz, CDCl3): δ 7.22 (d, J = 8.7 Hz, 2H), 6.90 (d, J = 8.7 Hz, 2H), 5.73 (ddt, J = 16.7, 10.4, 7.4 Hz, 1H), 5.18 - 5.03 (m, 3H), 4.54 - 4.38 (m, 2H), 3.91 (t, J = 5.5 Hz, 1H), 3.86 - 3.80 (m, 4H), 3.50 (d, J = 9.0 Hz, 1H), 3.41 (d, J = 9.2 Hz, 1H), 3.33 (d, J = 9.0 Hz, 1H), 3.09 (d, J = 6.5 Hz, 1H), 2.28 (ddt, J = 13.8, 7.4, 1.2 Hz, 1H), 2.11 (ddt, J = 13.8, 7.3, 1.2 Hz, 1H), 1.98 (dd, J = 14.2, 7.2 Hz, 1H), 1.52 - 1.40 (m, 1H) ppm. LCMS: [M+H] + m / z = 334.2 amu.

[0225] Preparation of intermediate 2-7

[0226] [ka] A flask containing intermediate 2-6 (70.0 mg, 1.0 equiv, 0.21 mmol) was charged with acetone:2,2'-dimethoxypropane (total 0.2 M, 10:1 mixture) and p-toluenesulfonic acid monohydrate (2.0 mg, 0.01 mmol, 5 mol%). The reaction was stirred at room temperature for 90 min until complete as determined by TLC. The solution was diluted with ethyl acetate (15 mL), transferred to a separatory funnel, and washed with sodium bicarbonate (10 mL). The organic portion was dried over sodium sulfate, filtered, and concentrated to give the crude acetonide, which did not require further purification (80.0 mg, crude, quantitative).

[0227] A mixture of the crude acetonide (185 mg, 1.0 equiv, 0.495 mmol) and water (0.09 mL, 10.0 equiv, 4.95 mmol) was treated with trimethylphosphine in THF (1.3 mL, 1 M, 2.5 equiv, 1.23 mmol). The reaction was stirred for 2 h and stripped of volatiles under a stream of nitrogen gas. After 1 h under high vacuum, the residue was dissolved in isopropyl alcohol (2 mL, 0.25 M) and Hunig's base (0.65 mL, 5 equiv, 3.72 mmol) was added followed by 2-(4,6-dichloropyrimidin-5-yl)acetaldehyde (142 mg, 1.5 equiv, 0.74 mmol). The reaction was sealed and heated to 75° C. for 12 h. LC / MS analysis indicated amine starting material remained. An additional and equal portion of Hunig's base and aldehyde was added and the reaction was resealed and heated to 85° C. for 5 h.

[0228] The reaction was poured into concentrated sodium bicarbonate and ethyl acetate. The organic phase was separated, dried over sodium sulfate, filtered, and concentrated to an oil which was purified by flash chromatography (0-100% ethyl acetate / hexanes). The product fractions were concentrated to dryness to give (1S,2R,3S,5R)-3-allyl-5-(4-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-(((4-methoxybenzyl)oxy)methyl)cyclopentane-1,2-diol (Intermediate 2-7, 137 mg, 58% yield) as a yellow oil. 1H NMR (400 MHz, CDCl3): δ 8.63 (s, 1H), 7.30 (d, J = 8.6 Hz, 2H), 7.24 (d, J = 3.6 Hz, 1H), 6.89 (d, J = 8.7 Hz, 2H), 6.60 (d, J = 3.6 Hz, 1H), 5.87 (dddd, J = 16.2, 10.8, 7.8, 6.9 Hz, 1H), 5.21 - 5.09 (m, 2H), 5.01 (ddt, J = 17.1, 7.7, 5.7 Hz, 2H), 4.62 (d, J = 6.8 Hz, 1H), 4.51 (s, 2H), 3.82 (s, 3H), 3.51 (s, 2H), 2.48 (ddd, J = 19.3, 13.8, 7.6 Hz, 2H), 2.33 - 2.17 (m, 2H), 1.52 (s, 3H), 1.32 - 1.15 (m, 3H) ppm. LCMS: [M+H] + m / z = 484.2 amu.

[0229] Alternative Preparation of Intermediate 2-3

[0230] [ka] Three reactions were carried out in parallel. To a solution of intermediate 2-2 (32.0 g, 117 mmol, 1.00 equiv.) in dichloromethane (300 mL), TEA (38.8 g, 383 mmol, 53.3 mL, 3.29 equiv.) and TMSOTf (28.4 g, 128 mmol, 23.1 mL, 1.10 equiv.) were added at -20 °C. The mixture was then stirred at 25 °C for 1 h. TLC (petroleum ether / acetone = 6 / 1, material R f =0.5, product R f=0.9) indicated the reaction was complete. The reaction mixture was quenched with saturated aqueous NaHCO3 (500 mL) at 0 °C and then extracted with dichloromethane (200 mL). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to give intermediate 2-2a (123 g, crude) as a yellow oil. The crude intermediate 2-2a was used in the next step without further purification.

[0231] [ka] Three reactions were carried out in parallel. To a mixture of intermediate 2-2a (41.0 g, 118 mmol, 1.00 equiv.) and allyl methyl carbonate (41.2 g, 355 mmol, 3.00 equiv.) in MeCN (250 mL) was added Pd2(dba)3 (3.25 g, 3.55 mmol, 0.03 equiv.). The mixture was stirred at 25 °C for 10 h. TLC (petroleum ether / acetone = 6 / 1, material R f =0.9, product R f =0.4) indicated the reaction was complete. The reaction mixtures were combined for workup. The combined mixture was filtered and washed with EtOAc (300 mL, 3 times). The filtrate was concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 30 / 1) to give intermediate 2-3 (53.0 g, 195 mmol, 54.8% yield) as a yellow oil. 1H NMR (400 MHz, CDCl3): δ 7.67 (dt, J = 5.6, 2.7 Hz, 1H), 7.23 - 7.12 (m, 2H), 6.93 - 6.78 (m, 2H), 6.16 (dt, J = 5.8, 2.2 Hz, 1H), 5.56 (dddd, J = 16.7, 10.0, 8.2, 6.5 Hz, 1H), 5.12 - 4.93 (m, 2H), 4.47 - 4.27 (m, 2H), 3.79 (s, 3H), 3.50 (d, J = 8.7 Hz, 1H), 3.36 (d, J = 8.7 Hz, 1H), 2.81 (dt, J = 19.2, 2.5 Hz, 1H), 2.54 (ddd, J = 19.2, 2.8, 2.1 Hz, 1H), 2.26 (ddt, J = 13.6, 6.5, 1.3 Hz, 1H), 2.18 (ddt, J = 13.5, 8.2, 1.0 Hz, 1H) ppm. LCMS: [M+Na] + m / z = 295.2 amu.

[0232] Alternative Preparation of Intermediate 2-4

[0233] [ka] Four reactions were carried out in parallel. To a solution of R-CBS (5.09 g, 18.4 mmol, 5.36 mL, 0.20 equiv.) and N,N-diethylaniline borane (21.0 g, 129 mmol, 22.9 mL, 1.40 equiv.) in THF (200 mL) was added intermediate 2-3 (25.0 g, 91.8 mmol, 1.00 equiv.) slowly under nitrogen atmosphere at -10 °C. The mixture was stirred at -10 °C for 1 h. TLC (petroleum ether / acetone = 6 / 1, material R f =0.4, product R f=0.35, PMA) indicated the reaction was complete. The four reactions were combined for workup. The reaction mixture was quenched with saturated aqueous NaHCO3 (1000 mL) and extracted with EtOAc (1000 mL, 2 times). The combined organic layers were washed with saturated aqueous NaHCO3 (2000 mL) and H2O (2000 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 30 / 1) to give intermediate 2-4 (73.0 g, 266 mmol, 72.5% yield) as a yellow oil. 1 H NMR (400 MHz, CDCl3): δ 7.39 - 7.17 (m, 2H), 6.92 - 6.84 (m, 2H), 5.83 (dtd, J = 5.8, 2.4, 1.0 Hz, 1H), 5.80 - 5.66 (m, 2H), 5.11 - 5.00 (m, 2H), 4.47 (dtd, J = 5.8, 2.2, 1.1 Hz, 1H), 4.44 (s, 1H), 3.81 (s, 3H), 3.49 (d, J = 1.2 Hz, 2H), 2.98 (d, J = 5.9 Hz, 1H), 2.34 - 2.20 (m, 2H), 2.20 - 2.07 (m, 2H) ppm. LCMS: [M+Na] + m / z = 297.1 amu.

[0234] Alternative Preparation of Intermediate 2-5

[0235] [ka] Two reactions were carried out in parallel. To a solution of intermediate 2-4 (25.0 g, 91.1 mmol, 1.00 equiv.) in DCM (250 mL) was added m-CPBA (27.8 g, 137 mmol, 85.0% purity, 1.50 equiv.) and NaHCO3 (19.1 g, 228 mmol, 8.86 mL, 2.50 equiv.) at 0 °C. The mixture was stirred at 0 °C for 6 h. TLC (petroleum ether / acetone = 6 / 1, material R f=0.35, product R f =0.2) indicated the reaction was complete. The reaction mixtures were combined for workup. The mixture was quenched with saturated aqueous Na2SO3 (3000 mL) and extracted with EtOAc (1000 mL, 3 times). The combined organic layers were washed with saturated aqueous NaHCO3 (3000 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the crude product. The crude product (Intermediate 2-5, 66.0 g, crude) was obtained as a colorless oil and was used in the next step without further purification. 1 H NMR (400 MHz, CDCl3): δ 7.25 - 7.19 (m, 2H), 6.90 - 6.84 (m, 2H), 5.70 (ddt, J = 16.8, 10.3, 7.4 Hz, 1H), 5.12 - 5.05 (m, 2H), 4.47 (d, J = 11.3 Hz, 1H), 4.34 (d, J = 11.4 Hz, 1H), 4.07 (s, 1H), 3.84 - 3.78 (m, 5H), 3.61 (dd, J = 2.9, 1.8 Hz, 1H), 3.42 (dd, J = 2.9, 1.8 Hz, 1H), 3.25 (d, J = 8.9 Hz, 1H), 2.53 - 2.41 (m, 1H), 2.12 (ddt, J = 13.6, 7.2, 1.2 Hz, 1H), 1.82 (d, J = 15.1 Hz, 1H), 1.69 (dd, J = 15.1, 1.9 Hz, 1H) ppm. LCMS: [M+H] + m / z = 291.2 amu.

[0236] Alternative Preparation of Intermediate 2-6

[0237] [ka] Two reactions were carried out in parallel. To a solution of intermediate 2-5 (33.0 g, 114 mmol, 1.00 equiv) in EtOH (500 mL) in H2O (100 mL) was added NaN3 (22.2 g, 341 mmol, 3.00 equiv) and NH4Cl (18.2 g, 341 mmol, 3.00 equiv). The mixture was stirred at 75 °C for 10 h. Then NaN3 (5.00 g, 76.9 mmol, 6.77e-1 equiv) was added and the mixture was stirred at 75 °C for 12 h. HPLC (intermediate 2-6: RT = 2.110 min, starting material: RT = 1.978 min) showed that most of the starting material was consumed. The two batches were combined for workup. The reaction mixture was diluted with H2O (2000 mL) and extracted with EtOAc (1000 mL, 3 times). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 10 / 1, TLC: petroleum ether / ethyl acetate = 1 / 1, product R f =0.75) to give intermediate 2-6 (21.0 g, 63.0 mmol, 27.7% yield) as a colorless oil. 1 H NMR (400 MHz, CDCl3): δ 7.22 (d, J = 8.7 Hz, 2H), 6.90 (d, J = 8.7 Hz, 2H), 5.73 (ddt, J = 16.7, 10.4, 7.4 Hz, 1H), 5.18 - 5.03 (m, 3H), 4.54 - 4.38 (m, 2H), 3.91 (t, J = 5.5 Hz, 1H), 3.86 - 3.80 (m, 4H), 3.50 (d, J = 9.0 Hz, 1H), 3.41 (d, J = 9.2 Hz, 1H), 3.33 (d, J = 9.0 Hz, 1H), 3.09 (d, J = 6.5 Hz, 1H), 2.28 (ddt, J = 13.8, 7.4, 1.2 Hz, 1H), 2.11 (ddt, J = 13.8, 7.3, 1.2 Hz, 1H), 1.98 (dd, J = 14.2, 7.2 Hz, 1H), 1.52 - 1.40 (m, 1H) ppm. LCMS: [M+H] + m / z = 334.2 amu.

[0238] Alternative Preparation of Intermediate 2-7

[0239] [ka] To a solution of intermediate 2-6 (21.0 g, 63.0 mmol, 1.00 equiv) in acetone (300 mL) was added TsOH·HO (599 mg, 3.15 mmol, 0.05 equiv) and 2,2'-dimethoxypropane (65.6 g, 630 mmol, 77.2 mL, 10.0 equiv). The mixture was stirred at 25 °C for 2 h. TLC (petroleum ether / ethyl acetate = 1 / 1, material R f =0.75, product R f =1, I2) indicated the reaction was complete. The reaction mixture was diluted with saturated aqueous NaHCO3 (10 mL) and H2O (200 mL) and then extracted with EtOAc (200 mL, 3 times). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to give the crude product (Intermediate 2-6a). Intermediate 2-6a (23.0 g, crude) was obtained as a yellow oil and used in the next step without further purification.

[0240] To a solution of intermediate 2-6a (23.0 g, 61.6 mmol, 1.00 equiv.) in THF (50.0 mL) was added PMe3 (1 M, 154 mL, 2.50 equiv.) and HO (11.1 g, 616 mmol, 11.1 mL, 10.0 equiv.). The mixture was stirred at 25 °C for 2 h. TLC (petroleum ether / ethyl acetate = 5 / 1, material R f =0.8, product R f = 0.3, I2) indicated the reaction was complete. The mixture was concentrated under high vacuum to give the crude product (Intermediate 2-6b). Intermediate 2-6b (25.0 g, crude) was obtained as a yellow oil and used in the next step without further purification.

[0241] To a solution of intermediate 2-6b (20.0 g, 57.6 mmol, 1.00 equiv) and 2-(4,6-dichloropyrimidin-5-yl)acetaldehyde (27.5 g, 144 mmol, 2.50 equiv) in i-PrOH (300 mL) was added TEA (29.1 g, 288 mmol, 40.1 mL, 5.00 equiv). The mixture was stirred at 75° C. for 36 h. LCMS showed the desired material, TLC (petroleum ether / ethyl acetate=5 / 1, material R f =0.3, product R f = 0.6, I2) showed the formation of one major spot. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 10 / 1) to give intermediate 2-7 (13.3 g, 26.6 mmol, yield 46.2%, purity 96.8%) as a yellow oil. Intermediate 2-6b: 1 H NMR (400 MHz, CDCl3): δ 7.27-7.30 (m, 2H), 6.86-6.93 (m, 2H), 5.69-5.93 (m, 1H), 5.01-5.16 (m, 2H), 4.23-4.56 (m, 4H), 3.83 (s, 3 H), 3.24-3.49 (m, 3H), 1.79-2.59 (m, 4H), 1.57 (s, 3H), 1.53 (s, 3H) ppm. Intermediate 2-7: 1H NMR (400 MHz, CDCl3): δ 8.63 (s, 1H), 7.30 (d, J = 8.6 Hz, 2H), 7.24 (d, J = 3.6 Hz, 1H), 6.89 (d, J = 8.7 Hz, 2H), 6.60 (d, J = 3.6 Hz, 1H), 5.87 (dddd, J = 16.2, 10.8, 7.8, 6.9 Hz, 1H), 5.21 - 5.09 (m, 2H), 5.01 (ddt, J = 17.1, 7.7, 5.7 Hz, 2H), 4.62 (d, J = 6.8 Hz, 1H), 4.51 (s, 2H), 3.82 (s, 3H), 3.51 (s, 2H), 2.48 (ddd, J = 19.3, 13.8, 7.6 Hz, 2H), 2.33 - 2.17 (m, 2H), 1.52 (s, 3H), 1.32 - 1.15 (m, 3H) ppm. LCMS: [M+H] + m / z = 484.2 amu.

[0242] Preparation of Intermediate 2-8 and Epi-Intermediate 2-8

[0243] [ka] A flask containing intermediate 2-7 (449 mg, 1.0 equiv, 0.93 mmol) was charged with THF (3 mL), water (1 mL) and 2,6-dimethylpyridine (0.22 mL) followed by gentle heating to obtain a homogeneous solution. Solid sodium periodate (794 mg, 4.0 equiv, 3.71 mmol) was added followed by osmium tetroxide (4% solution in water, 0.18 mL, 3 mol%, 0.03 mmol) and the reaction was stirred at ambient temperature for 2 h. TLC analysis showed complete conversion and the reaction was poured into 1N aqueous sodium sulfite solution (2 volumes) followed by dilution with ethyl acetate (4 volumes). The aqueous phase was separated and the organic phase was further washed with concentrated sodium bicarbonate. The resulting organic phase was dried over sodium sulfate, filtered, concentrated to dryness and used directly in the following step.

[0244] To prepare the Grignard reagent, a dry flask was charged with benzyl (3-bromo-7-iodoquinolin-2-yl)(4-methoxybenzyl)carbamate (710 mg, 1.27 equiv, 1.18 mmol) and dry THF (8 mL) was injected to give a homogenous solution. After cooling to -78 °C, isopropylmagnesium chloride solution (2 M in THF, 0.584 mL, 1.26 equiv, 1.17 mmol) was injected. The reaction was stirred for 1 h and then allowed to warm to ambient temperature. LC / MS confirmed the iodine-magnesium exchange and the solution was re-cooled to -78 °C.

[0245] The crude aldehyde prepared above was dissolved in dry THF (8 mL) and cooled to -78 °C, and the Grignard reagent prepared above was transferred to the reaction flask via cannula. The resulting homogenous solution was stirred for 1 h and assessed by LC / MS, which indicated consumption of the aldehyde. The cooling bath was removed and the reaction was warmed for 5 min and then poured into concentrated ammonium chloride solution (50 mL). The reaction flask was then washed with ethyl acetate (150 mL). After vigorous stirring, the mixture was transferred to a separatory funnel and the organic phase was separated. The organic phase was dried over sodium sulfate, filtered, concentrated to dryness, and purified by reverse phase HPLC (30 mm column, 50-100% acetonitrile / water + 0.25% acetic acid, 254 nM detection, 12 injections). Two fractions were collected (fraction 1 corresponding to intermediate 2-8, 250 mg, 28% yield; fraction 2 corresponding to Epi-intermediate 2-8, 200 mg, 23% yield; combined yield for two steps: 51%), with fraction 1 eluting first and fraction 2 eluting second. Fraction 1 (Intermediate 2-8): 1 H NMR (400 MHz, acetonitrile-d3, 70℃): δ 8.59 (s, 1H), 8.48 (s, 1H), 7.89 (s, 1H), 7.81 (d, J = 8.5 Hz, 1H), 7.65 (dd, J = 8.5, 1.6 Hz, 1H), 7.59 (d, J = 3.7 Hz, 1H), 7.37 (d, J = 8.6 Hz, 2H), 7.34 - 7.23 (m, 7H), 6.98 - 6.89 (m, 2H), 6.80 (d, J = 8.7 Hz, 2H), 6.66 (d, J = 3.7 Hz, 1H), 5.44 (s, 1H), 5.23 - 5.08 (m, 3H), 4.99 (d, J = 4.8 Hz, 3H), 4.59 (dd, J = 15.8, 6.5 Hz, 3H), 3.76 (dd, J = 21.6, 10.5 Hz, 6H), 3.55 (d, J = 4.2 Hz, 1H), 2.57 (dd, J = 13.5, 7.8 Hz, 1H), 2.42 - 2.33 (m, 1H), 2.12 - 2.06 (m, 2H), 1.77 (p, J = 2.4 Hz, 1H), 1.47 (s, 3H), 1.15 (s, 3H) ppm. LCMS: [M+H] + m / z = 962.3 amu. Draw points 2 (Epi-intermediate 2-8): 1 H NMR (400 MHz, アセトトトリル-d3, 70℃): δ 1H NMR (400 MHz, acetonitrile-d3) δ 8.58 (s, 1H), 8.52 - 8.40 (m, 1H), 7.96 - 7.83 (m, 1H), 7.77 (dd, J = 8.5, 1.7 Hz, 1H), 7.64 - 7.51 (m, 2H), 7.43 - 7.19 (m, 8H), 7.02 - 6.85 (m, 2H), 6.86 - 6.73 (m, 2H), 6.73 - 6.60 (m, 1H), 5.44 (s, 2H), 5.22 - 5.13 (m, 3H), 5.05 (dd, J = 7.3, 5.5 Hz, 1H), 5.00 (s, 3H), 4.86 (d, J = 7.3 Hz, 1H), 4.55 (s, 2H), 3.78 (s, 3H), 3.73 (s, 3H), 2.60 (dd, J = 13.2, 7.6 Hz, 1H), 2.25 (dd, J = 13.3, 11.5 Hz, 1H), 2.12 (dd, J = 14.5, 3.1 Hz, 1H), 2.07 - 1.98 (m, 2H), 1.51 (s, 3H), 1.28 (s, 3H) ppm. LCMS: [M+H] + m / z = 962.3 amu.

[0246] Preparation of intermediate 2-8a

[0247] [ka] A flask containing Intermediate 2-8 and Epi-Intermediate 2-8 (approximately 1:1 mixture, 12.98 g, 13.47 mmol) was dissolved in DCM (250 mL) and solid Dess-Martin periodinane (6.86 g, 16.17 mmol) was added in one portion. The reaction was stirred for 3 h, at which point the conversion was deemed complete by TLC analysis (4:1 DCM / EtOAc). The reaction mixture was diluted with 50% saturated sodium sulfite (200 mL) and stirred for 30 min. Volatiles were removed by evaporation and diethyl ether (500 mL) was added.

[0248] The mixture was transferred to a separatory funnel and the organic phase was further washed with 10% sodium bicarbonate (100 mL, twice) and saturated sodium bicarbonate (100 mL). The aqueous phase was further washed with additional diethyl ether (200 mL) and the combined organic washes were dried over magnesium sulfate, then filtered and concentrated to give a white foam (12.95 g, quantitative yield) which was used without further purification.

[0249] The isolated ketone was dissolved in dry DCM (120 mL) and solid RuCl[(S,S)-Tsdpen](p-cymene) was added (800 mg, 0.0933 mmol) under a nitrogen atmosphere. Formic acid-triethylamine azeotrope (6.18 g, 5.30 mmol) was added and the reaction was stirred at ambient temperature for 4 days. At this point, LC / MS analysis indicated complete conversion and the reaction was concentrated onto silica gel.

[0250] Flash chromatography (0-30% EtOAc / DCM) afforded intermediate 2-8a after concentration to a white foam (10.37 g, 10.765 mmol, 80% yield over two steps). NMR analysis showed a diastereoselectivity of >20:1 for the process.

[0251] Intermediate 2-8a can be carried forward in the same manner as described for the mixture of intermediate 2-8 and Epi-intermediate 2-8 to provide the biologically active stereoisomers without separation of the isomeric mixture. 1H NMR (400 MHz, アセトニトリル-d3): δ 8.61 (s, 1H), 8.52 (s, 1H), 7.88 (s, 1H), 7.84 (d, J = 8.5 Hz, 1H), 7.70 - 7.63 (m, 2H), 7.43 - 7.36 (m, 2H), 7.36 - 7.24 (m, 8H), 6.99 - 6.91 (m, 2H), 6.87 - 6.78 (m, 2H), 6.69 (d, J = 3.7 Hz, 1H), 5.18 (s, 3H), 5.13 - 5.06 (m, 1H), 5.00 (s, 2H), 4.63 - 4.51 (m, 2H), 3.82 - 3.66 (m, 9H), 2.56 (dd, J = 13.4, 7.6 Hz, 1H), 2.00 (s, 6H) ppm. LCMS: [M+H] + m / z = 962.2 amu.

[0252] Modulation of Intermediate 2-9 and Epi-Intermediate 2-9

[0253]

change

[0254] The diol (95 mg, 0.11 mmol) was placed under a nitrogen atmosphere (3× high vacuum / nitrogen cycles) and a solution of (cyanomethylene)trimethylphosphorane (0.5 M, Sigma-Aldrich, 1.8 mL, 0.90 mmol) was injected. The vessel was sealed and warmed to 50° C., and the reaction was deemed complete within 4 h by LC / MS analysis. The reaction was quenched with methanol, transferred onto silica gel, and concentrated to dryness. Flash chromatography (0–50% EtOAc / DCM) afforded intermediate 2-9 as a white solid after concentration of the product fractions (78 mg, 84% yield). 1H NMR (400 MHz, acetonitrile-d3): δ 8.60 (s, 1H), 8.50 (s, 1H), 7.90 - 7.76 (m, 2H), 7.64 - 7.53 (m, 2H), 7.38 - 7.17 (m, 7H), 6.84 - 6.75 (m, 2H), 6.67 (d, J = 3.7 Hz, 1H), 5.22 - 5.01 (m, 2H), 4.97 (s, 2H), 4.70 (d, J = 6.2 Hz, 1H), 4.48 (d, J = 8.8 Hz, 1H), 3.81 (d, J = 8.8 Hz, 1H), 3.72 (s, 3H), 2.58 (dd, J = 13.4, 6.8 Hz, 1H), 2.51 - 2.43 (m, 1H), 2.37 (dd, J = 12.5, 6.9 Hz, 1H), 2.03 (dd, J = 12.6, 9.0 Hz, 1H), 1.54 (s, 3H), 1.26 (s, 3H) ppm. LCMS: [M+H] + m / z = 824.2 amu.

[0255] Preparation of intermediate 2-9a

[0256] [ka] Intermediate 2-9a was prepared using the same procedure used to prepare intermediate 2-9 and was obtained as a white solid (6.40 g, 78% yield). 1H NMR (400 MHz, acetonitrile-d3): δ 8.60 (s, 1H), 8.50 (s, 1H), 7.90 - 7.76 (m, 2H), 7.64 - 7.53 (m, 2H), 7.38 - 7.17 (m, 7H), 6.84 - 6.75 (m, 2H), 6.67 (d, J = 3.7 Hz, 1H), 5.22 - 5.01 (m, 4H), 4.97 (s, 2H), 4.70 (d, J = 6.2 Hz, 1H), 4.48 (d, J = 8.8 Hz, 1H), 3.81 (d, J = 8.8 Hz, 1H), 3.72 (s, 3H), 2.58 (dd, J = 13.4, 6.8 Hz, 1H), 2.51 - 2.43 (m, 1H), 2.37 (dd, J = 12.5, 6.9 Hz, 1H), 2.03 (dd, J = 12.6, 9.0 Hz, 1H), 1.54 (s, 3H), 1.34 - 1.19 (s, 3H) ppm. LCMS: [M+H] + m / z = 824.2 amu.

[0257] Preparation of Intermediate 2-10 and Epi-Intermediate 2-10

[0258] [ka] A flask containing intermediate 2-7 (1.0 equiv, 0.93 mmol) is charged with THF (3 mL), water (1 mL) and 2,6-dimethylpyridine (0.22 mL) followed by gentle heating to obtain a homogeneous solution. Solid sodium periodate (4.0 equiv, 3.71 mmol) is added followed by osmium tetroxide (4% solution in water, 0.18 mL, 3 mol%, 0.03 mmol) and the reaction is stirred at ambient temperature for 2 h. The reaction is then poured into 1N aqueous sodium sulfite solution (2 volumes) followed by dilution with ethyl acetate (4 volumes). The aqueous phase is separated and the organic phase is further washed with concentrated sodium bicarbonate. The resulting organic phase is dried over sodium sulfate, filtered, concentrated to dryness and used directly in the subsequent step.

[0259] To prepare the Grignard reagent, a dry flask is charged with benzyl (3-bromo-7-iodoquinolin-2-yl)(4-methoxybenzyl)carbamate (1.27 equiv., 1.18 mmol) and dry THF (8 mL) is injected to obtain a homogeneous solution. After cooling to -78 °C, an isopropylmagnesium chloride solution (2 M in THF, 0.584 mL, 1.26 equiv., 1.17 mmol) is injected. The reaction is stirred for 1 h and then allowed to warm to ambient temperature. The solution is then re-cooled to -78 °C.

[0260] The crude aldehyde prepared above is dissolved in dry THF (8 mL) and cooled to -78 °C, and the Grignard reagent prepared above is transferred to the reaction flask via cannula. The resulting homogeneous solution is stirred for 1 h. The cooling bath is then removed, and the reaction is warmed for 5 min and then poured into concentrated ammonium chloride solution (50 mL). The reaction flask is then washed with ethyl acetate (150 mL). After vigorous stirring, the mixture is transferred to a separatory funnel and the organic phase is separated. The organic phase is dried over sodium sulfate, filtered, concentrated to dryness, and purified by reverse-phase HPLC (30 mm column, 50-100% acetonitrile / water + 0.25% acetic acid, 254 nM detection, 12 injections). Two fractions are collected (fraction 1 corresponding to intermediate 2-10, fraction 2 corresponding to Epi-intermediate 2-10), with fraction 1 eluting first and fraction 2 eluting second.

[0261] Preparation of Intermediate 2-11 and Epi-Intermediate 2-11

[0262] [ka] Epi-intermediate 2-10 is also subjected to the following general procedure to generate the epimer of intermediate 2-11. Intermediate 2-10 (1.0 equiv, 0.12 mmol) is dissolved in 1:1 acetonitrile / dichloromethane (5 mL). A reaction vial is charged with a solution of DDQ (1.3 equiv, 0.15 mmol) in pH 7 phosphate buffer (0.34 mL) and an equal volume of acetonitrile / dichloromethane (1:1, 5 mL), the mixture is transferred into the aqueous buffer solution, and the flask is cooled to 0° C. The substrate is transferred into the vial via pipette and stirred at 0° C. for 2 h. The reaction is removed from the ice bath and stirred at ambient temperature for 5 h. The reaction is poured into 0.2 N sodium carbonate (100 mL) and the aqueous phase is washed three times with DCM (50 mL portions). The combined organic washes are dried over sodium sulfate, filtered, concentrated onto silica gel, and purified by flash chromatography (0-100% DCM / EtOAc) to give the diol.

[0263] The diol (0.11 mmol) is placed under an atmosphere of nitrogen (3x high vacuum / nitrogen cycles) and a solution of (cyanomethylene)trimethylphosphorane (0.5 M, Sigma-Aldrich, 1.8 mL, 0.90 mmol) is injected. The vessel is sealed and warmed to 50 °C. The reaction is quenched with methanol, transferred onto silica gel, and concentrated to dryness. Flash chromatography (0-50% EtOAc / DCM) gives intermediate 2-11.

[0264] Preparation of Compounds 11 and 12

[0265] [ka] Epi-intermediate 2-11 was also subjected to the following general procedure to produce compound 12, the epimer of compound 11. Intermediate 2-11 (1.0 equiv.) was dissolved in methanol (0.03 M) to give a solution, followed by dissolution of 4N hydrochloric acid in dioxane (10 equiv.). The reaction was stirred at room temperature until complete as determined by LC / MS analysis, concentrated to dryness, and the residue was dissolved in DMSO.

[0266] After filtration, reverse phase HPLC is performed (acetonitrile / water + 0.25% acetic acid, 20 mm x 250 mm C18 column, 40 mL / min gradient, 242 nM detection wavelength, 3 injections total). The product fractions are pooled, frozen and concentrated on a lyophilizer to give compound 11.

[0267] Preparation of Compounds 13 and 14

[0268] [ka] Epi-intermediate 2-11 was also subjected to the following general procedure to produce compound 14, the epimer of compound 13. A vial of intermediate 2-11 (1.0 equiv.) was treated with dioxane (0.1 M) to give a clear solution, followed by ammonium hydroxide (0.1 M). The reaction was stirred at 100° C. until complete, cooled, and the solvent was concentrated in vacuo.

[0269] To the crude material is added methanol (0.03M) and 4N hydrochloric acid in dioxane (10 equivalents). Upon completion as determined by LC / MS analysis, the solution is concentrated, dissolved in DMSO, filtered, and purified by reverse phase HPLC (acetonitrile / water + 0.25% acetic acid, 20 mm x 250 mm C18 column, 40 mL / min gradient over 15 min, detection wavelength of 242 nM). Product fractions are pooled, frozen and concentrated on a lyophilizer to give compound 13.

[0270] Preparation of Compounds 15 and 16

[0271] [ka] Epi-intermediate 2-9 was also subjected to the following general procedure to produce compound 16. Intermediate 2-9 (45 mg, 1.0 equiv, 0.05 mmol) was placed in a microwave vial and dioxane was added (1.36 mL) followed by ammonium hydroxide (1.36 mL). The mixture became homogeneous upon heating to 100° C. and intermediate 2-9 was consumed over 16 h with the formation of the ammonia nucleophilic displacement product. The solvent was then stripped, first under a stream of nitrogen and then under high vacuum. TFA was then injected (1.37 mL) followed immediately by thioanisole (0.13 mL). The reaction was sealed and heated to 80° C. for a period of 75 min, at which point LC / MS analysis showed compound 15 and its corresponding trifluoroacetate salt. The solvent was removed under a stream of nitrogen and then under high vacuum for 1 h. The residue was dissolved in methanol (2 mL) and potassium carbonate was added (75.4 mg, 10.0 equiv, 0.55 mmol). After stirring at ambient temperature for 1 h, the mixture was filtered through a pad of Celite and all contact surfaces were washed with copious amounts of ethyl acetate and methanol (10:1). After concentration, the residue was dissolved in DMSO containing a small amount of acetic acid, filtered, and purified by reverse phase HPLC (30 mm, 45 mL / min, 254 nM, 5-25% acetonitrile / water + 0.25% acetic acid). The product fractions were frozen and concentrated to dryness on a lyophilizer to give compound 15 as a free base form. Fraction 1: 1H NMR (400 MHz, Acetonitrile-d3 + 3 vol% d-TFA): δ 8.62 (s, 1H), 8.13 (s, 1H), 7.76 (d, J = 8.4 Hz, 1H), 7.62 (d, J = 1.7 Hz, 1H), 7.45 (dd, J = 8.5, 1.4 Hz, 1H), 7.41 (d, J = 3.7 Hz, 1H), 6.81 (d, J = 3.7 Hz, 1H), 5.14 - 4.96 (m, 2H), 4.39 (dd, J = 8.4, 4.4 Hz, 1H), 4.33 (d, J = 8.8 Hz, 1H), 3.84 (d, J = 4.4 Hz, 1H), 3.74 (d, J = 8.9 Hz, 1H), 2.56 (dd, J = 12.3, 6.6 Hz, 1H), 2.48 (dd, J = 13.9, 10.2 Hz, 1H), 2.05 - 2.00 (m, 2H) ppm. LCMS: [M+H] + m / z = 511.1 amu. Drawing points 2: 1 H NMR (400 MHz, Acetonitrile-d3 + 3 vol% d-TFA): δ 8.58 (s, 1H), 8.19 (s, 1H), 7.77 - 7.70 (m, 2H), 7.44 - 7.39 (m, 2H), 6.84 (dd, J = 3.7, 1.0 Hz, 1H), 5.15 (m, 1H), 4.97 (m, 1H), 4.66 (m, 1H), 3.79 (m, 1H), 2.64 - 2.60 (m, 2H), 2.05 - 2.00 (m, 2H) ppm. LCMS: [M+H] + m / z = 511.1 amu.

[0272] Preparation of compounds 17 and 18

[0273]

change

[0274] Preparation of the mesylate salt of compound 15

[0275] [ka] The free base of compound 15 was suspended in water and methanesulfonic acid was added (32 mg in 2 mL of water). The resulting solution was gently heated and sonicated until a homogeneous solution was obtained, which was then frozen and concentrated to dryness in a lyophilizer. The resulting oily solid was redissolved in water, frozen and concentrated in the same manner to give a water-soluble white solid (approximately 3 mg / mL). Further experiments were more easily carried out with this salt (32.0 mg, 91% yield for two steps). 1 H NMR (400 MHz, heavy water) δ 8.69 (d, J = 0.9 Hz, 1H), 8.23 ​​(d, J = 0.7 Hz, 1H), 7.84 (d, J = 8.3 Hz, 1H), 7.68 - 7.59 (m, 1H), 7.58 - 7.46 (m, 2H), 6.88 (d, J = 3.6 Hz, 1H), 5.29 - 5.12 (m, 2H), 4.61 (dd, J = 9.2, 4.3 Hz, 1H), 4.45 (d, J = 8.9 Hz, 1H), 4.05 (d, J = 4.3 Hz, 1H), 3.91 (d, J = 9.0 Hz, 1H), 2.68 - 2.58 (m, 2H), 2.24 - 2.02 (m, 2H) ppm. LC / MS: [M+H] + m / z = 511.1 amu.

[0276] Preparation of compound 19

[0277] [ka] A vial was charged with tetrakis(triphenylphosphine)palladium(0) (35 mg, 0.0303 mmol) and intermediate 2-9a and dissolved in THF (3 mL). MeAl (2 M in toluene, 0.33 mL) was then injected and the reaction was heated at 60° C. for 2 h. The reaction was determined to be complete by LC / MS analysis and was cooled to ambient temperature, at which point 1 mL of methanol was added.

[0278] The solvent was removed under a stream of nitrogen gas followed by vacuum. The residue was dissolved in TFA (5 mL) and thioanisole (5 eq, 0.20 mL) was injected followed by heating to 65° C. for 1 h. LC / MS analysis indicated that the conversion was not complete so an additional 5 mL of TFA was injected and the reaction was heated to 80° C. in a sealed vessel for an additional 1 h, after which LC / MS analysis indicated the reaction was complete.

[0279] The solvent was removed under a stream of nitrogen gas followed by vacuum, and the residue was dissolved in MeOH (3 mL) and then K2CO3 (3 equiv, 140 mg) in H2O (1 mL) was injected. The reaction was stirred for 1 h, then neutralized with 0.5 mL of AcOH and filtered. The filtrate was then purified by HPLC (10-20% H2O / CH3CN+0.25% AcOH).

[0280] The combined HPLC fractions were concentrated by evaporation to 50%, and 2M aqueous Na2CO3 (25 mL) was added to precipitate the product, which was collected by filtration. The filtrate was lyophilized to dryness and transferred to a vial to give compound 19 (113 mg, 0.2214 mmol, 65.64% yield) as a white solid. 1H NMR (400 MHz, acetonitrile-d3+ 1% d-TFA): δ 8.93 (s, 1H), 8.61 (s, 1H), 7.84 - 7.67 (m, 3H), 7.50 (dd, J = 8.3, 1.6 Hz, 1H), 7.03 (d, J = 3.8 Hz, 1H), 5.31 - 5.08 (m, 2H), 4.54 (dd, J = 8.7, 4.3 Hz, 1H), 4.43 (d, J = 8.9 Hz, 1H), 3.92 (d, J = 4.2 Hz, 1H), 3.80 (d, J = 8.9 Hz, 1H), 2.93 (s, 3H), 2.69 (dd, J = 12.3, 6.5 Hz, 1H), 2.58 (dd, J = 14.0, 10.4 Hz, 1H), 2.21 (dd, J = 14.0, 8.4 Hz, 1H) ppm. LC / MS: [M+H] + m / z = 510.1 amu.

[0281] Preparation of Compounds 20 and 21

[0282] [ka] A microwave vial was charged with intermediate 2-9a (12.4 mg, 0.015 mmol) and palladium on carbon (10%, 4 mg) and charged with methanol under a nitrogen atmosphere. The reaction was placed under hydrogen gas (balloon, atmospheric pressure) and monitored by LC / MS.

[0283] After 2 hours, additional palladium on carbon (10%, 4 mg) was added and hydrogenation was continued for 2 hours. The reaction was filtered and concentrated. The filtrate was dissolved in TFA (1 mL) and thioanisole (50 μL, 5% by volume) was injected. The reaction was sealed and heated to 70° C. for 1 hour.

[0284] The solvent was removed under a stream of nitrogen. The residue was dissolved in MeOH (1 mL) and stirred with 2N aqueous K2CO3 (50 μL, 5% by volume) for 1 h. The reaction was filtered, and the filtrate was washed with MeOH (1 mL) and purified by HPLC (20 mm, 23 mL / min, 10-30% water / ACN+0.25% TFA buffer). The active fractions were frozen and lyophilized to give compounds 20 and 21. Compound 20: 1 H NMR (400 MHz, acetonitrile-d3): δ 8.51 (d, J = 1.5 Hz, 1H), 8.17 (dt, J = 9.2, 1.0 Hz, 1H), 7.76 (dd, J = 8.4, 1.5 Hz, 1H), 7.62 - 7.51 (m, 2H), 7.41 (dd, J = 8.2, 1.7 Hz, 1H), 6.97 - 6.83 (m, 1H), 6.68 - 6.51 (m, 1H), 5.31 - 4.88 (m, 2H), 4.56 - 4.44 (m, 1H), 4.36 (dd, J = 8.8, 1.5 Hz, 1H), 3.83 (d, J = 4.3 Hz, 1H), 3.73 (dd, J = 8.9, 1.5 Hz, 1H), 2.65 (dd, J = 12.4, 6.7 Hz, 1H), 2.49 (dd, J = 13.9, 10.4 Hz, 1H), 2.20 - 2.04 (m, 1H) ppm. LC / MS: [M+H] + m / z = 452.1 amu. Compound 21: 1H NMR (400 MHz, acetonitrile-d3): δ 9.00 (s, 1H), 8.86 (s, 1H), 8.15 - 8.09 (m, 1H), 7.74 (d, J = 8.3 Hz, 1H), 7.69 (d, J = 3.7 Hz, 1H), 7.62 (d, J = 1.5 Hz, 1H), 7.38 (dd, J = 8.3, 1.6 Hz, 1H), 6.88 (d, J = 9.3 Hz, 1H), 6.81 (d, J = 3.7 Hz, 1H), 5.17 - 5.04 (m, 2H), 4.46 (dd, J = 8.6, 4.3 Hz, 1H), 4.36 (d, J = 8.9 Hz, 1H), 3.83 (d, J = 4.3 Hz, 1H), 3.71 (d, J = 8.9 Hz, 1H), 2.61 (dd, J = 12.3, 6.6 Hz, 1H), 2.49 (dd, J = 13.9, 10.4 Hz, 1H), 2.15 - 2.10 (m, 2H) ppm. LC / MS: [M+H] + m / z = 418.2 amu.

[0285] Preparation of compound 22

[0286] [ka] A microwave vial was charged with intermediate 2-9a (22 mg, 0.0267 mmol) and tetrakis(triphenylphosphine)palladium(0) (5 mg, 0.0043 mmol) and charged with THF (0.3 mL) under argon, followed by triethylsilane (5 μL) and after 5 h the reaction was filtered and concentrated.

[0287] The filtrate was dissolved in TFA (1 mL) and thioanisole (50 μL, 5% by volume) was injected. The reaction was sealed and heated to 70° C. for 1 h. The solvent was then removed under a stream of nitrogen gas followed by vacuum. The crude residue was dissolved in MeOH (1 mL) and stirred with 2N aqueous K2CO3 (50 μL, 5% by volume) for 1 h.

[0288] The mixture was filtered, and the filtrate was washed with MeOH (1 mL) and purified by HPLC (20 mm, 23 mL / min, 10-30% water / ACN+0.25% TFA buffer). The active fraction was frozen and lyophilized to give compound 22 (5.0 mg, 37% yield) as a white solid. 1 H NMR (400 MHz, acetonitrile-d3): δ 8.98 (s, 1H), 8.83 (s, 1H), 8.53 (s, 1H), 7.73 - 7.63 (m, 3H), 7.42 (dd, J = 8.3, 1.5 Hz, 1H), 6.81 (dd, J = 3.7, 1.0 Hz, 1H), 5.20 - 5.00 (m, 2H), 4.47 (dd, J = 8.6, 4.3 Hz, 1H), 4.36 (d, J = 8.9 Hz, 1H), 3.83 (d, J = 4.3 Hz, 1H), 3.73 (d, J = 8.8 Hz, 1H), 2.55 - 2.43 (m, 3H), 2.13 (dd, J = 13.9, 8.4 Hz, 1H) ppm. LC / MS: [M+H] + m / z = 496.1 amu.

[0289] Preparation of compound 23 Compound 23 was synthesized using intermediate 2-9a as the starting material and the corresponding amine nucleophile (2N methylamine in methanol) by the general procedure used for compound 15. Compound 23 was obtained as a white solid. 1H NMR (500 MHz, methanol-d4): δ 8.34 (d, J = 2.9 Hz, 1H), 8.19 (s, 1H), 7.71 - 7.60 (m, 2H), 7.36 (dt, J = 8.4, 1.9 Hz, 1H), 7.29 (dd, J = 3.7, 1.5 Hz, 1H), 6.63 (d, J = 3.5 Hz, 1H), 5.20 - 5.10 (m, 2H), 4.61 (dd, J = 8.3, 4.4 Hz, 1H), 4.56 (d, J = 8.8 Hz, 1H), 4.02 (d, J = 4.4 Hz, 1H), 3.89 (d, J = 7.6 Hz, 1H), 3.11 (s, 3H), 2.73 - 2.61 (m, 2H), 2.18 (dd, J = 13.4, 8.3 Hz, 1H), 2.10 - 2.04 (m, 1H) ppm. LCMS: [M+H] + m / z = 525.1 amu.

[0290] Preparation of compound 24 Compound 24 was synthesized using intermediate 2-9a as the starting material and the corresponding amine nucleophile (2N ethylamine in methanol) by the general procedure used for compound 15. Compound 24 was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6): δ 8.53 (s, 1H), 8.25 (s, 1H), 7.67 (d, J = 8.3 Hz, 1H), 7.58 (s, 1H), 7.49 (s, 1H), 7.30 - 7.20 (m, 1H), 6.90 - 6.80 (m, 1H), 5.05 - 4.92 (m, 2H), 4.32 (dd, J = 8.8, 4.0 Hz, 1H), 4.25 (d, J = 8.7 Hz, 1H), 3.70 (d, J = 4.0 Hz, 1H), 3.60 (d, J = 8.8 Hz, 1H), 2.51 (dd, J = 12.1, 6.3 Hz, 1H), 2.35 (dd, J = 13.8, 10.5 Hz, 1H), 1.92 - 1.70 (m, 2H), 1.19 (t, J = 7.2 Hz, 3H) ppm. LCMS: [M+H] + m / z = 539.1 amu.

[0291] Preparation of compound 25 Compound 25 was synthesized using intermediate 2-9a as the starting material and the corresponding amine nucleophile (isopropylamine) by the general procedure used for compound 15. Compound 25 was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6): δ 8.54 (s, 1H), 8.26 (s, 1H), 7.69 (d, J = 8.3 Hz, 1H), 7.59 (s, 1H), 7.51 (s, 1H), 7.30 - 7.22 (m, 1H), 6.96 - 6.87 (m, 1H), 5.08 - 4.89 (m, 2H), 4.34 (dd, J = 8.8, 4.0 Hz, 1H), 4.26 (d, J = 8.8 Hz, 1H), 4.20 - 4.10 (m, 1H), 3.72 (d, J = 4.0 Hz, 1H), 3.61 (d, J = 8.8 Hz, 1H), 2.52 (dd, J = 12.2, 6.3 Hz, 1H), 2.41 - 2.31 (m, 1H), 1.89 (dd, J = 13.7, 8.1 Hz, 1H), 1.79 (dd, J = 12.2, 9.6 Hz, 1H), 1.24 (d, J = 6.4 Hz, 6H) ppm. LCMS: [M+H] + m / z = 553.1 amu.

[0292] Preparation of compound 26 Compound 26 was synthesized using intermediate 2-9a as the starting material and the corresponding amine nucleophile (n-propylamine) by the general procedure used for compound 15. Compound 26 was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6): δ 8.54 (s, 1H), 8.27 (s, 1H), 7.69 (d, J = 8.3 Hz, 1H), 7.64 - 7.40 (m, 2H), 7.27 (dd, J = 8.3, 1.6 Hz, 1H), 6.99 - 6.73 (m, 1H), 5.07 - 4.92 (m, 2H), 4.40 - 4.30 (m, 1H), 4.26 (d, J = 8.8 Hz, 1H), 3.72 (d, J = 4.0 Hz, 1H), 3.61 (d, J = 8.8 Hz, 1H), 2.52 (dd, J = 12.2, 6.3 Hz, 1H), 2.37 (dd, J = 13.8, 10.6 Hz, 1H), 1.96 - 1.73 (m, 2H), 1.60 (q, J = 7.3 Hz, 2H), 0.90 (t, J = 7.4 Hz, 3H) ppm. LCMS: [M+H] + m / z = 553.1 amu.

[0293] [Example 3] Synthesis of other compounds General Route 1

[0294] [ka] Using this general approach, various compounds herein can be synthesized. Introduction of various groups at the R position is achieved by adapting the starting allylic 2-oxocyclopentane-1-carboxylate compound prior to decarboxylative enantioselective allylic alkylation. Introduction of the nucleoside can then be achieved by heterocyclyl or cycloalkyl cyclization followed by nucleophilic addition with Grignard or other organometallic reagents, or nucleophiles.

[0295] In other embodiments, the compounds of the invention have nucleosides containing spiro-carbocycles or spiro-heterocyclyls, the ring size being 5-6 members, which may also be synthesized by this general approach. The heteroatoms in the spiro-heterocyclyl are selected from oxygen, nitrogen and sulfur. The spiro ring may have any of the configurations and substitution patterns described herein. In a preferred embodiment, the R group is an optionally substituted aryl or heteroaryl, with one, two or three optional substituents independently selected from H, F, Cl, Br, methyl, NH2, N(CH3)2, OH or O(CH3).

[0296] General Route 2

[0297] [ka] Alternatively, the approach of General Route 1 is not limited to the use of decarboxylative enantioselective allylic alkylation. Beta-ketoester building blocks can be used in place of various nucleophiles in nucleophilic addition chemistry. Specifically, beta-ketoesters can undergo nucleophilic addition to unsaturated nitro compounds. After reduction and spontaneous lactam formation, compounds bearing lactams at the indicated positions in the scheme of General Route 2 can be obtained.

[0298] General Route 3

[0299] [ka] The carbonyl of the compound obtained in general route 2 can be replaced by modification of the aryl fragment. A ketone can be introduced by Negishi coupling of benzyl (3-bromo-7-iodoquinolin-2-yl) (4-methoxybenzyl) carbamate with 2-tert-butoxy-2-oxoethylzinc bromide. The ester is converted to its corresponding Mander's reagent and reacted in the presence of lithium enolate of cyclopentane to generate a new 1,3-dicarbonyl species known to participate in the Mannich chemistry mentioned by Y. Numajiri, BP Pritchett, K. Chiyoda and BM Stoltz, published in J. Am. Chem. Soc. (2015), Vol. 137, pp. 1040-1043. From here, selective bromination and cyclization of the benzyl ketone generates the corresponding bicyclic species, which are easily converted to their corresponding nucleoside analogues.

[0300] Biological Experiments PRMT5 enzyme inhibition assay A chemiluminescence immunoassay (CLIA) to measure the amount of PRMT5-mediated histone methylation was performed in a 96-well plate format according to the manufacturer's protocol (BPS Biosciences, Cat#52002L). 96-well plates precoated with histone H4 peptide were rehydrated prior to use. A master mix containing the PRMT5 substrate S-adenosylmethionine, reaction buffer, and purified PRMT5 enzyme was added to the ready wells. Wells without either the S-adenosylmethionine substrate or the PRMT5 enzyme served as negative controls. Ten test concentrations of test compounds were prepared, ranging from 100 μM to 10 nM, using a 1:3 serial dilution in dH2O. Five microliters of each test concentration was added to the ready wells. Five microliters of 1% DMSO in water without inhibitor was added to the negative control wells and also to the positive control wells to serve as ready wells. The final reaction volume in each well was 50 μL, and the final concentrations of test compounds in each well ranged from 10 μM to 1 nM. EPZ015666, a known inhibitor of PRMT5, was tested in parallel reaction wells as an assay validation control. The 96-well plate was incubated at room temperature for 1 hour.

[0301] After incubation was completed, the reaction supernatant from each well was discarded. The wells were washed with 1xTBST and incubated in blocking buffer for 1 hour. The wells were incubated with a primary antibody that recognizes the methylated Arg3 residue of histone H4 for 1 hour. The wells were washed again with 1xTBST and then incubated with an HRP-conjugated secondary antibody for 30 minutes. The wells were washed again with 1xTBST and an equal part of chemiluminescence reagent was added to each well. Luminescence was quantified on a plate reader.

[0302] Luminescence values ​​were normalized to positive and negative controls and data were converted to %PRMT5 activity. Inhibitor concentration-dependence curves were plotted against %PRMT5 activity and IC values ​​were calculated for each compound. 50 IC values ​​were determined. See Table 3. "A" is an IC of 100 nM or less. 50 "B" stands for IC of 101nM to 500nM. 50 "C" stands for IC > 500nM50 Represents.

[0303] Cell line proliferation delay assay Cells were seeded at a density of 1,000-5,000 cells per well in 48-well tissue culture plates. After a 24-hour resting period, cells were treated with compounds at 1 μM, 0.2 μM, 0.04 μM, 0.008 μM, 0.0016 μM, and 0.00032 μM. One group of cells was treated with the vehicle in which the compounds were prepared and served as a control. Cells were counted prior to treatment and this count was used as a baseline for calculation of growth inhibition. Cells were grown in the presence of compounds for 6 days and counted on the sixth day. All cell counts were performed using a Synentec Cellavista plate imager. Growth inhibition was calculated as the ratio of cell population doublings in the presence of compound versus the absence of compound and expressed as the maximum percentage growth inhibition achieved over the range of compound tested concentrations ("maximum effect" or "ME"), calculated by fitting curves to the data points from each dose-response assay using the Proc NLIN function in SAS for Windows, version 9.2 (SAS Institute, Inc.).

[0304] Assignment of sensitive and resistant cohorts and calculation of mean maximum effect size Human cancer cell lines were grouped as "sensitive" or "resistant" to PRMT5 inhibition based on whether their growth was delayed by JNJ-64619178 (i.e., (1S,2R,3R,5R)-3-(2-(2-amino-3-bromoquinolin-7-yl)ethyl)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopentane-1,2-diol) (data not shown, see Table 4). These sensitive and resistant cohorts were examined for response to each compound and ME was calculated for each cell line using the same technique described above. The average ME for the sensitive and resistant cohorts was calculated as the arithmetic mean of the group. See Table 3. "A" represents a percent maximal effect of 60% or more, "B" represents a percent maximal effect of 30% to 59%, and "C" represents a percent maximal effect of 29% or less.

[0305] Caco-2 assay (P app A B) The extent of bidirectional human intestinal permeability of compounds was estimated using a Caco-2 cell permeability assay. Caco-2 cells were seeded on polyethylene membranes in 96-well plates. Growth medium was refreshed every 4-5 days until the cells formed a confluent cell monolayer. HBSS containing 10 mM HEPES at pH 7.4 was used as the transport buffer. Compounds were tested in duplicate at 2 μM in both directions. Digoxin, nadolol and metoprolol were included as standards. Digoxin was tested in duplicate at 10 μM in both directions, while nadolol and metoprolol were tested in duplicate at 2 μM in the AB direction. Final DMSO concentrations were adjusted to less than 1% for all experiments. Plates were incubated for 2 hours in a CO2 incubator at 37°C with saturated humidity and 5% CO2. After incubation, all wells were mixed with acetonitrile containing the internal standard and the plates were centrifuged at 4,000 rpm for 10 minutes. 100 μL of supernatant was collected from each well and diluted with 100 μL of distilled water for LC / MS / MS analysis. The concentrations of test and control compounds in the starting, donor and receiver solutions were quantified by LC / MS / MS using the analyte / internal standard peak area ratios.

[0306] The apparent permeability coefficient, P app (cm / s) was calculated using the following formula: P app =(dC r / dt) × V r / (A×C0) [In the formula, dC r / dt is the cumulative concentration of compound in the receiver chamber as a function of time (μM / s); V r is the solution volume in the receiver chamber (0.075 mL on the apical side and 0.25 mL on the basolateral side); A is the surface area for transport, which is 0.0804 cm for the area of ​​the monolayer. 2 and C0 is the initial concentration in the donor chamber (μM). appThe scores are shown in Table 3. "A" represents a score of 1 or greater, "B" represents a score between 0.2 and 0.99, and "C" represents a score less than 0.2.

[0307] The discharge ratio was calculated using the following formula: Emission ratio=P app (BA) / P app (AB)

[0308] The percent recovery was calculated using the following formula: Recovery rate (%) = 100 × [(V r ×C r )+(V d ×C d )] / (V d ×C0) [where Vd is the volume in the donor chamber, which is 0.075 mL on the apical side and 0.25 mL on the basolateral side; C d and C r are the final concentrations of transport compound in the donor and receiver chambers, respectively].

[0309] Determining the metabolic stability of compounds The metabolic stability of the compounds was determined in hepatocytes from human, mouse and rat. Compounds were diluted to 5 μM in Williams' E medium from a 10 mM stock solution. 10 μL of each compound was aliquoted into wells of a 96-well plate, and the reaction was initiated by aliquoting 40 μL of a 625,000 cell / mL suspension into each well. The plate was incubated at 37° C. with 5% CO2. At each corresponding time point, the reaction was stopped by quenching with ACN containing an internal standard (IS) at 1:3. The plate was shaken at 500 rpm for 10 minutes and then centrifuged at 3,220×g for 20 minutes. The supernatant was transferred to another 96-well plate containing the dilution solution. The supernatant was analyzed by LC / MS / MS. Half-life (min) and CL of exemplary compounds measured in mouse hepatocytes int Value (μL / min / 10 6 The half-life of the CL cells is shown in Table 3. With respect to half-life, "A" indicates more than 50 minutes, "B" indicates 15 to 50 minutes, and "C" indicates less than 15 minutes. intRegarding "A", 100μL / min / 10 6 "B" stands for 50-100 μL / min / 10 6 "C" stands for 50 μL / min / 10 6 Represents less than a cell.

[0310] The percent of compound remaining after incubation was calculated using the following formula: Remaining compound (%) = Peak area ratio of test compound to internal standard at the end point / Peak area ratio of test compound to internal standard at the start point

[0311] Compound half-life and CL int was calculated using the following formula: C t =C0*e -k*t (first order kinetics);C t If = 1 / 2C0, t 1 / 2 = ln2 / k = 0.693 / k; and CL int =k / (1,000,000 cells / mL)

[0312] Activity-guided selection of inhibitors A subgenus of PRMT5 inhibitors with desirable properties was identified using a combination of in vitro data.

[0313] In particular, the above-mentioned assays (e.g., cell line proliferation delay assay, PRMT5 enzyme inhibition assay, Caco-2 assay (P app Results from the study (AB), measurement of the metabolic stability of the compounds, and assignment of sensitive and resistant cohorts and calculation of the mean maximum effect) were used to select compounds having the structural and functional characteristics defined in the subgenus of formula (V).

[0314] In particular, as described above, desirable properties of compounds tested in sensitive and resistant cell lines are that they have an average ME against drug-sensitive cell lines of Table 4 of at least 60% or more, and an average ME against drug-resistant cell lines of Table 4 of about 60% or less.

[0315] One of skill in the art will readily recognize that the results of additional in vitro assays (e.g., CYP enzyme inhibition, hERG inhibition, compound solubility, target specificity analysis), as well as the results of in vivo assays (e.g., rodent xenograft studies, rodent pharmacokinetic and single dose saturation studies, rodent maximum tolerated dose studies, and oral bioavailability) may be used to identify other subgenuses of PRMT5 inhibitors, or to narrow the subgenus determined using other results, such as that of Formula (V).

[0316] Examples of Estimated PRMT5 Binding Affinities of Exemplary Compounds The structure of compound 15 bound to PRMT5:MEP50 was determined by X-ray crystallography.

[0317] [ka]

[0318] The Molecular Operating Environment (MOE) software package (Chemical Computing Group, Montreal, Canada) version 2019.0102 was used to generate conformational ensembles of example compounds by flexible alignment to the central core structure of compound 15 seen in the X-ray structure. This provided an ensemble of energetically achievable conformers of all substituents of interest.

[0319] Energy minimization and estimated approximate binding affinity in the crystal structure of PRMT5:MEP50 were determined using two methods: (i) running GB-IV using the AMBER10:EHT force field in the MOE software package described above ("Method 1"), and (ii) running MMGB-SA using the OPLS3e force field in the Prime module of the Schrodinger software suite (v. 2020-1, Schrodinger LLC, New York, NY) ("Method 2").

[0320] In some embodiments, compounds of the invention have a predicted binding affinity, as determined by one of these methods, that is better than the predicted binding affinity of compound 15. In preferred embodiments, compounds of the invention have a predicted binding affinity, as determined by both of these methods, that is better than the predicted binding affinity of compound 15.

[0321] The estimated binding affinities of exemplary compounds determined by each method are shown in Table 5. Values ​​are in arbitrary units ("AU"), with more negative values ​​corresponding to higher estimated affinities. The estimated binding affinity of compound 15 using method 1 is -8.84 and -15.08 using method 2.

[0322] [Table 3]

[0323] [Table 4] TIFF0007609798000160.tif146161

[0324] [Table 5] TIFF0007609798000162.tif251152TIFF0007609798000163.tif253152TIFF0007609798000164.tif225153TIFF0007609798000165.tif99161 (Additional Note) (Appendix 1) Formula I: [ka] or a pharma- ceutically acceptable salt thereof. [In the formula, [ka] is a double bond or a single bond, Ar is an optionally substituted aryl or heteroaryl, where one, two or three optional substituents are halogen, hydroxyl, cyano, -C(O)NH, -C(O)N(R a )2, -C(O)O(R a ), -C(O)R a , -N(R a )2, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl, optionally substituted C1-C4 alkyl, and optionally substituted C2-C6 alkenyl; R1 is H, halogen, C1-C4 alkyl, C2-C4 alkenyl, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl or N(R a )2, X1 and X2 are each independently CH or N; Y1 is (CH2) n or C(O), with the proviso that when Y2 or Y3 is C(O), then Y1 cannot be C(O); Y2 is CH2, C(O), S, SO2, O or NR a However, when Y1 or Y3 is C(O), Y2 cannot be C(O); [ka] is a single bond, Y3 is CH2 or C(O), provided When Y1 or Y2 is C(O), Y3 cannot be C(O); [ka] is a single bond, or Y2 and Y3 are CH; [ka] is a double bond, Z is CH2, O, S or NH; R ais independently, at each occurrence, H, C1-C4 alkyl, or C3-C6 cycloalkyl; m is 0, 1, 2 or 3; n is 1 or 2]. (Appendix 2) Formula Ia: [ka] or a pharma- ceutically acceptable salt thereof. (Appendix 3) Formula Ib: [ka] or a pharma- ceutically acceptable salt thereof. (Appendix 4) Formula II: [ka] or a pharma- ceutically acceptable salt thereof. [In the formula, [ka] is, independently in each occurrence, a double bond or a single bond; R1 is H, halogen, C1-C4 alkyl, C2-C4 alkenyl, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl or N(R a )2, R2 is H, halogen, hydroxyl, cyano, -C(O)NH2, -C(O)N(R a )2, -C(O)O(R a ), -C(O)R a , -N(R a )2, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl, optionally substituted C1-C4 alkyl or optionally substituted C2-C6 alkenyl; W is C(R b) or C(O), provided that when W is C(O), V is NH, and [ka] is a single bond, V is CH, NH, or N, provided that when V is CH or N, W is C(R b ) between W and V [ka] is a double bond, X1 and X2 are each independently CH or N; Y1 is (CH2) n or C(O), with the proviso that when Y2 or Y3 is C(O), then Y1 cannot be C(O); Y2 is CH2, C(O), S, SO2, O or NR a However, when Y1 or Y3 is C(O), Y2 cannot be C(O); Between Y2 and Y3 [ka] is a single bond, Y3 is CH2 or C(O), provided When Y1 or Y2 is C(O), Y3 cannot be C(O); Between Y2 and Y3 [ka] is a single bond, or Y2 and Y3 are CH; [ka] is a double bond, Z is CH2, O, S or NH; R a is independently, at each occurrence, H, C1-C4 alkyl, or C3-C6 cycloalkyl; R bis H, halogen, hydroxyl, cyano, -C(O)NH2, -C(O)N(R a )2, -C(O)O(R a ), -C(O)R a , -N(R a )2, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl, optionally substituted C1-C4 alkyl or optionally substituted C2-C6 alkenyl; m is 0, 1, 2 or 3; n is 1 or 2]. (Appendix 5) Formula IIa: [ka] or a pharma- ceutically acceptable salt thereof. (Appendix 6) Formula IIb: [ka] or a pharma- ceutically acceptable salt thereof. (Appendix 7) Formula III: [ka] or a pharma- ceutically acceptable salt thereof. [In the formula, Ar is an optionally substituted aryl or heteroaryl, where one, two or three optional substituents are halogen, hydroxyl, cyano, -C(O)NH, -C(O)N(R a )2, -C(O)O(R a ), -C(O)R a , -N(R a )2, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl, optionally substituted C1-C4 alkyl, and optionally substituted C2-C6 alkenyl; R1 is H, halogen, C1-C4 alkyl, C2-C4 alkenyl, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl or N(R a )2, X1 and X2 are each independently CH or N; Y2 is CH2, S, O or N(R a ) and Z is CH2, O, S or NH; R a is independently, at each occurrence, H, C1-C4 alkyl, or C3-C6 cycloalkyl; m is 0, 1, 2 or 3. (Appendix 8) Formula IIIa: [ka] or a pharma- ceutically acceptable salt thereof. (Appendix 9) Formula IIIb: [ka] or a pharma- ceutically acceptable salt thereof. (Appendix 10) Formula IV: [ka] or a pharma- ceutically acceptable salt thereof. [In the formula, R1 is H, halogen, C1-C4 alkyl, C2-C4 alkenyl, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl or N(R a )2, R2 is H, halogen, hydroxyl, cyano, -C(O)NH2, -C(O)N(R a )2, -C(O)O(R a ), -C(O)R a , -N(R a )2, -O(R a), C3-C6 cycloalkyl, C3-C7 heterocyclyl, optionally substituted C1-C4 alkyl or optionally substituted C2-C6 alkenyl; X1 and X2 are each independently CH or N; V is CH or N; Y2 is CH2, S, O or N(R a ) and Z is CH2, O, S or NH; R a is independently, at each occurrence, H, C1-C4 alkyl, or C3-C6 cycloalkyl; R b is H, halogen, hydroxyl, cyano, -C(O)NH2, -C(O)N(R a )2, -C(O)O(R a ), -C(O)R a , -N(R a )2, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl, optionally substituted C1-C4 alkyl or optionally substituted C2-C6 alkenyl. (Appendix 11) Formula IVa: [ka] or a pharma- ceutically acceptable salt thereof. (Appendix 12) Formula IVb: [ka] or a pharma- ceutically acceptable salt thereof. (Appendix 13) Formula V: [ka] or a pharma- ceutically acceptable salt thereof. [In the formula, R1 is H, halogen, C1-C4 alkyl, C2-C4 alkenyl, -O(Ra ), C3-C6 cycloalkyl, C3-C7 heterocyclyl or N(R a )2, R2 is H, halogen, hydroxyl, cyano, -C(O)NH2, -C(O)N(R a )2, -C(O)O(R a ), -C(O)R a , -N(R a )2, -O(R a ), C3-C6 cycloalkyl, C3-C7 heterocyclyl, optionally substituted C1-C4 alkyl or optionally substituted C2-C6 alkenyl; X2 is CH or N; Y2 is CH2, S, O or N(R a ) and R a is independently, at each occurrence, H, C1-C4 alkyl, or C3-C6 cycloalkyl. wherein the compound has a PRMT5 IC of about 100 nM or less. 50 or a pharma- ceutically acceptable salt thereof. (Appendix 14) Compounds described in Appendix 13 having an average percent maximal efficacy against drug-resistant cell lines of Table 4 of about 60% or less. (Appendix 15) A compound according to claim 13 or 14 having an average percent maximum effect of about 60% or more against a drug-sensitive cell line of Table 4. (Appendix 16) Formula Va: [ka] or a pharma- ceutically acceptable salt thereof. (Appendix 17) Formula Vb: [ka] or a pharma- ceutically acceptable salt thereof. (Appendix 18) The compound according to any one of claims 1, 4, 7, or 10, wherein X1 is CH. (Appendix 19) The compound according to any one of claims 1 to 17, wherein X2 is CH. (Appendix 20) The compound according to any one of claims 1 to 17, wherein X2 is N. (Appendix 21) The compound of any one of claims 4, 5, 6, 10, 11, or 12, wherein V is N. (Appendix 22) R b is Cl or NH2. (Appendix 23) R b The compound according to any one of claims 4, 5, 6, 10, 11, or 12, wherein is NH2. (Appendix 24) The compound of any one of claims 1 to 12 or 18 to 23, wherein Z is CH2. (Appendix 25) The compound according to any one of claims 1 to 24, wherein Y2 is N(H). (Appendix 26) The compound according to any one of claims 1 to 24, wherein Y2 is O. (Appendix 27) The compound according to any one of claims 1 to 26, wherein R1 is Cl or NH2. (Appendix 28) The compound according to any one of claims 1 to 26, wherein R1 is NH2. (Appendix 29) The compound according to any one of claims 4 to 6 or 10 to 28, wherein R2 is Cl or Br. (Appendix 30) The compound according to any one of claims 4 to 6 or 10 to 28, wherein R2 is Br. (Appendix 31) X1 is CH, X2 is CH, V is N, and R bA compound according to any one of claims 4 to 7, wherein: is NH2, Z is CH2, Y2 is N(H), R1 is Cl or NH2, and R2 is Br. (Appendix 32) X1 is CH, X2 is CH, V is N, and R b is NH2, Z is CH2, Y2 is N(H) or O, R1 is Cl or NH2, and R2 is Br. (Appendix 33) [ka] TIFF0007609798000192.tif251154TIFF0007609798000193.tif215161TIFF0007609798000194.tif216159TIFF0007609798000195.tif208159TIFF0007609798000196.tif207160TIFF0007609798000197.tif249155TIFF0007609798000198.tif99149, or a pharma- ceutically acceptable salt thereof. (Appendix 34) [ka] TIFF0007609798000200.tif199163, or a pharma- ceutically acceptable salt thereof. (Appendix 35) [ka] or a pharma- ceutically acceptable salt thereof. (Appendix 36) [ka] or a pharma- ceutically acceptable salt thereof. (Appendix 37) [ka] or a pharma- ceutically acceptable salt thereof. (Appendix 38) [ka] or a pharma- ceutically acceptable salt thereof. (Appendix 39) [ka] or a pharma- ceutically acceptable salt thereof. (Appendix 40) [ka] or a pharma- ceutically acceptable salt thereof. (Appendix 41) [ka] or a pharma- ceutically acceptable salt thereof. (Appendix 42) [ka] or a pharma- ceutically acceptable salt thereof. (Appendix 43) [ka] TIFF0007609798000210.tif249162TIFF0007609798000211.tif252161TIFF0007609798000212. tif230165TIFF0007609798000213.tif209156TIFF0007609798000214.tif208153TIFF00076097 98000215.tif249162TIFF0007609798000216.tif205156TIFF0007609798000217.tif208161TIFF0007609798000218.tif247165TIFF0007609798000219.tif52151, or a pharma- ceutically acceptable salt thereof. (Appendix 44) [ka] TIFF0007609798000221.tif248153TIFF0007609798000222.tif252163TIFF0007609798000223.tif25116 1TIFF0007609798000224.tif251156TIFF0007609798000225.tif228163TIFF0007609798000226.tif2091 64TIFF0007609798000227.tif223157TIFF0007609798000228.tif248156TIFF0007609798000229.tif249165TIFF0007609798000230.tif225163TIFF0007609798000231.tif130164, or a pharma- ceutically acceptable salt thereof. (Appendix 45) [ka] TIFF0007609798000233.tif200156, or a pharma- ceutically acceptable salt thereof. (Appendix 46) [ka] TIFF0007609798000235.tif245161TIFF0007609798000236.tif157148, or a pharma- ceutically acceptable salt thereof. (Appendix 47) A compound selected from Table 2. (Appendix 48) A pharmaceutical composition comprising a compound according to any one of claims 1 to 47 and a pharma- ceutically acceptable diluent or excipient. (Appendix 49) A method of treating a disease characterized by increased expression of PRMT5 in a subject, comprising administering to a subject identified as in need of such treatment an effective amount of a compound according to any one of claims 1 to 47 or a pharma- ceutically acceptable salt thereof. (Appendix 50) 50. The method of claim 49, wherein the disease is selected from cancer, a metabolic disease, and a hematological disease. (Appendix 51) 51. The method of claim 49 or 50, wherein the disease is cancer. (Appendix 52) Cancers include carcinomas (e.g., carcinomas of the endometrium, bladder, breast, colon (e.g., colorectal carcinoma, e.g., colon adenocarcinoma and colon adenoma), sarcomas (e.g., Kaposi's sarcoma, osteosarcoma, tumors of mesenchymal origin, e.g., fibrosarcoma or rhabdomyosarcoma), kidney, epidermis, liver, lung (e.g., adenocarcinoma, small cell lung carcinoma and non-small cell lung carcinoma), esophagus, gallbladder, ovary, pancreas (e.g., exocrine pancreatic carcinoma), stomach, cervix, thyroid, nose, head and neck, prostate, and skin (e.g., squamous cell carcinoma), human breast cancer (e.g., primary breast tumors, node-negative breast cancer, invasive ductal adenocarcinoma of the breast, nonendometrioid breast cancer), familial melanoma, melanoma, hematopoietic tumors of the lymphoid lineage (e.g., leukemia, 52. The method of claim 51, wherein the tumor is selected from acute lymphocytic leukemia, mantle cell lymphoma, chronic lymphocytic leukemia, B-cell lymphoma (e.g., diffuse large B-cell lymphoma), T-cell lymphoma, multiple myeloma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma, and Burkitt's lymphoma), and hematopoietic tumors of the myeloid lineage, such as acute and chronic myeloid leukemia, myelodysplastic syndrome, promyelocytic leukemia, tumors of the central or peripheral nervous system, such as astrocytoma, neuroblastoma, glioma or schwannoma, seminoma, teratocarcinoma, xeroderma pigmentosum, retinoblastoma, keratoacanthoma, and follicular thyroid carcinoma. (Appendix 53) 53. The method of claim 51 or 52, wherein the cancer is selected from colorectal cancer, head and neck cancer, pancreatic cancer, sarcoma, melanoma, myeloma, lymphoma, lung cancer (including non-small cell lung cancer and small cell lung cancer), breast cancer, ovarian cancer, liver cancer, gastric cancer, endometrial cancer, renal cancer, bladder cancer, and acute myeloid leukemia. (Appendix 54) 54. The method of claim 53, wherein the cancer is selected from colorectal cancer, head and neck cancer, pancreatic cancer, sarcoma, melanoma, myeloma, lymphoma, non-small cell lung cancer, breast cancer, ovarian cancer, liver cancer, gastric cancer, bladder cancer, and acute myeloid leukemia. (Appendix 55) A method for inhibiting PRMT5 in a cell, comprising contacting the cell with a compound according to any one of appendix 1 to 47 or a pharma- ceutically acceptable salt thereof, such that PRMT5 is inhibited in the cell. (Appendix 56) 56. The method of claim 55, wherein the cell is a cancer cell. (Appendix 57) 57. The method of claim 55 or 56, wherein cell proliferation is inhibited, cell death is induced, or both.

Claims

1. Formula I: 【Chemistry 1】 or a pharma- ceutically acceptable salt thereof. [In the formula, 【Chemistry 2】 is a double bond or a single bond, Ar is an optionally substituted aryl or heteroaryl, wherein one, two or three optional substituents are halogen, hydroxyl, cyano, -C(O)NH 2 , -C(O)N(R a ) 2 , -C(O)O(R a ), -C(O)R a , -N(R a ) 2 , -O(R a ), C 3 ~C 6 Cycloalkyl, C 3 ~C 7 Heterocyclyl, optionally substituted C 1 ~C 4 Alkyl and optionally substituted C 2 ~C 6 alkenyl, R 1 H, halogen, C 1 ~C 4 Alkyl, C 2 ~C 4 Alkenyl, -O(R a ), C 3 ~C 6 Cycloalkyl, C 3 ~C 7 Heterocyclyl or N(R a ) 2 and X 1 and X 2 are each independently CH or N; Y 1 (CH 2 ) n or C(O), where Y 2 Or Y 3 If C(O), then Y 1 cannot be C(O), Y 2 CH 2 , C(O), S, SO 2 , O or NR a However, Y 1 Or Y 3 If is C(O), then Y 2 cannot be C(O), 【Chemistry 3】 is a single bond, Y 3 CH 2 or C(O), where Y 1 Or Y 2 If C(O), then Y 3 cannot be C(O), 【Chemistry 4】 is a single bond, or Y 2 and Y 3 is CH, 【Chemistry 5】 is a double bond, Z is CH 2 , O, S or NH; R a are, in each case independently, H, C 1 ~C 4 Alkyl or C 3 ~C 6 is cycloalkyl, m is 0, 1, 2 or 3; n is 1 or 2].

2. Formula Ia: 【Chemistry 6】 2. The compound of claim 1 having the structure:

3. Formula II: 【Chemistry 7】 or a pharma- ceutically acceptable salt thereof. [In the formula, 【Chemistry 8】 is, independently in each occurrence, a double bond or a single bond; R 1 H, halogen, C 1 ~C 4 Alkyl, C 2 ~C 4 Alkenyl, -O(R a ), C 3 ~C 6 Cycloalkyl, C 3 ~C 7 Heterocyclyl or N(R a ) 2 and R 2 is H, halogen, hydroxyl, cyano, -C(O)NH 2 , -C(O)N(R a ) 2 , -C(O)O(R a ), -C(O)R a , -N(R a ) 2 , -O(R a ), C 3 ~C 6 Cycloalkyl, C 3 ~C 7 Heterocyclyl, optionally substituted C 1 ~C 4 Alkyl or optionally substituted C 2 ~C 6 alkenyl, W is C(R b ) or C(O), provided that when W is C(O), V is NH, and 【Chemistry 9】 is a single bond, V is CH, NH, or N, provided that when V is CH or N, W is C(R b ) between W and V 【Chemistry 10】 is a double bond, X 1 and X 2 are each independently CH or N; Y 1 (CH 2 ) n or C(O), where Y 2 Or Y 3 If C(O), then Y 1 cannot be C(O), Y 2 CH 2 , C(O), S, SO 2 , O or NR a However, Y 1 Or Y 3 If is C(O), then Y 2 cannot be C(O), Y 2 and Y 3 Between 【Chemistry 11】 is a single bond, Y 3 CH 2 or C(O), where Y 1 Or Y 2 If C(O), then Y 3 cannot be C(O), Y 2 and Y 3 Between 【Chemistry 12】 is a single bond, or Y 2 and Y 3 is CH and Y 2 and Y 3 Between 【Chemistry 13】 is a double bond, Z is CH 2 , O, S or NH; R a are, in each case independently, H, C 1 ~C 4 Alkyl or C 3 ~C 6 is cycloalkyl, R b is H, halogen, hydroxyl, cyano, -C(O)NH 2 , -C(O)N(R a ) 2 , -C(O)O(R a ), -C(O)R a , -N(R a ) 2 , -O(R a ), C 3 ~C 6 Cycloalkyl, C 3 ~C 7 Heterocyclyl, optionally substituted C 1 ~C 4 Alkyl or optionally substituted C 2 ~C 6 alkenyl, m is 0, 1, 2 or 3; n is 1 or 2].

4. Formula IIa: 【Chemistry 14】 4. The compound of claim 3 having the structure: or a pharma- ceutically acceptable salt thereof.

5. Formula III: 【Chemistry 15】 or a pharma- ceutically acceptable salt thereof. [In the formula, Ar is an optionally substituted aryl or heteroaryl, wherein one, two or three optional substituents are halogen, hydroxyl, cyano, -C(O)NH 2 , -C(O)N(R a ) 2 , -C(O)O(R a ), -C(O)R a , -N(R a ) 2 , -O(R a ), C 3 ~C 6 Cycloalkyl, C 3 ~C 7 Heterocyclyl, optionally substituted C 1 ~C 4 Alkyl and optionally substituted C 2 ~C 6 alkenyl, R 1 H, halogen, C 1 ~C 4 Alkyl, C 2 ~C 4 Alkenyl, -O(R a ), C 3 ~C 6 Cycloalkyl, C 3 ~C 7 Heterocyclyl or N(R a ) 2 and X 1 and X 2 are each independently CH or N; Y 2 CH 2 , S, O or N(R a ) and Z is CH 2 , O, S or NH; R a are, in each case independently, H, C 1 ~C 4 Alkyl or C 3 ~C 6 is cycloalkyl, m is 0, 1, 2 or 3.

6. Formula IIIa: 【Chemistry 16】 6. The compound of claim 5 having the structure: or a pharma- ceutically acceptable salt thereof.

7. Formula IV: 【Chemistry 17】 or a pharma- ceutically acceptable salt thereof. [In the formula, R 1 H, halogen, C 1 ~C 4 Alkyl, C 2 ~C 4 Alkenyl, -O(R a ), C 3 ~C 6 Cycloalkyl, C 3 ~C 7 Heterocyclyl or N(R a ) 2 and R 2 is H, halogen, hydroxyl, cyano, -C(O)NH 2 , -C(O)N(R a ) 2 , -C(O)O(R a ), -C(O)R a , -N(R a ) 2 , -O(R a ), C 3 ~C 6 Cycloalkyl, C 3 ~C 7 Heterocyclyl, optionally substituted C 1 ~C 4 Alkyl or optionally substituted C 2 ~C 6 alkenyl, X 1 and X 2 are each independently CH or N; V is CH or N; Y 2 CH 2 , S, O or N(R a ) and Z is CH 2 , O, S or NH; R a are, in each case independently, H, C 1 ~C 4 Alkyl or C 3 ~C 6 is cycloalkyl, R b is H, halogen, hydroxyl, cyano, -C(O)NH 2 , -C(O)N(R a ) 2 , -C(O)O(R a ), -C(O)R a , -N(R a ) 2 , -O(R a ), C 3 ~C 6 Cycloalkyl, C 3 ~C 7 Heterocyclyl, optionally substituted C 1 ~C 4 Alkyl or optionally substituted C 2 ~C 6 alkenyl].

8. Formula IVa: 【Chemistry 18】 8. The compound of claim 7 having the structure: or a pharma- ceutically acceptable salt thereof.

9. Formula Va: 【Chemistry 19】 or a pharma- ceutically acceptable salt thereof. [In the formula, R 1 H, halogen, C 1 ~C 4 Alkyl or N(R a ) 2 and R 2 is H or a halogen, X 2 is CH, Y 2 is O, R a is, independently in each occurrence, H or C 1 ~C 4 is alkyl] and the compound has a PRMT5 IC of 100 nM or less. 50 or a pharma- ceutically acceptable salt thereof. 【Request 10】 【Chemical 20】 10. The compound of claim 9, having a structure selected from:

11. X 1 8. The compound of claim 1, 3, 5 or 7, wherein is CH.

12. X 2 9. The compound according to claim 1, wherein is CH.

13. 9. The compound of claim 3, 4, 7, or 8, wherein V is N.

14. R b NH 2 9. The compound of claim 3, 4, 7, or 8, wherein

15. Z is CH 2 9. The compound according to any one of claims 1 to 8, wherein

16. Y 2 9. The compound according to claim 1, wherein is N(H).

17. Y 2 The compound according to any one of claims 1 to 8, wherein is O.

18. R 1 is Cl or NH 2 9. The compound according to any one of claims 1 to 8, wherein

19. R 2 10. The compound according to any one of claims 3 to 4 or 7 to 9, wherein is Br.

20.

21. 【change】 【change】 【change】 【change】 【change】 【change】 【change】 or a pharma- ceutically acceptable salt thereof.

21.

22. 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 or a pharma- ceutically acceptable salt thereof.

22.

23. 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 or a pharma- ceutically acceptable salt thereof.

23.

24. 【change】 【change】 or a pharma- ceutically acceptable salt thereof.

24. 24. A pharmaceutical composition comprising a compound according to any one of claims 1 to 23 and a pharma- ceutically acceptable diluent or excipient.

25. 24. Use of a compound according to any one of claims 1 to 23, or a pharma- ceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of a disease characterized by increased expression of PRMT5.

26. the disease is selected from cancer, metabolic disease, and blood disease; 26. The use according to claim 25, wherein the cancer is selected from carcinoma, sarcoma, tumors of mesenchymal origin, renal cancer, epidermal cancer, liver cancer, lung cancer, esophageal cancer, gallbladder cancer, ovarian cancer, pancreatic cancer, gastric cancer, cervical cancer, thyroid cancer, nasal cancer, head and neck cancer, prostate cancer, skin cancer, human breast cancer, familial melanoma, melanoma, hematopoietic tumors of lymphoid lineage, hematopoietic tumors of myeloid lineage, tumors of the central or peripheral nervous system, seminoma, teratocarcinoma, xeroderma pigmentosum, retinoblastoma, keratoacanthoma, and follicular thyroid cancer.

27. The use of claim 26, wherein the cancer is selected from endometrial, bladder, breast, colon carcinoma, colon adenocarcinoma, colon adenoma, Kaposi's sarcoma, osteosarcoma, fibrosarcoma, rhabdomyosarcoma, adenocarcinoma, small cell and non-small cell lung cancer, exocrine pancreatic carcinoma, squamous cell carcinoma, primary breast tumors, node-negative breast cancer, invasive ductal adenocarcinoma of the breast, non-endometrioid breast cancer, leukemia, acute lymphocytic leukemia, mantle cell lymphoma, chronic lymphocytic leukemia, B-cell lymphoma, diffuse large B-cell lymphoma, T-cell lymphoma, multiple myeloma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma, Burkitt's lymphoma, acute and chronic myeloid leukemia, myelodysplastic syndromes, promyelocytic leukemia, astrocytoma, neuroblastoma, glioma, and schwannoma.

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