Piperidinyl-methyl-purineamines as NSD2 inhibitors and Anti-cancer agents

Abstract

To provide a compound, composition, and method for inhibiting nuclear SET domain-containing protein 2 (NSD2).SOLUTION: Disclosed is a compound of Formula (I) or an enantiomer, an enantiomeric mixture, or a pharmaceutically acceptable salt thereof. Further disclosed are a pharmaceutical composition comprising such compounds, and methods of using such compounds for treating a disease or condition mediated by nuclear SET domain-containing protein 2 (NSD2).SELECTED DRAWING: None

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of PCT / CN2019 / 100542, filed August 14, 2019, which is incorporated by reference in its entirety.

[0002] The present invention relates to compounds, compositions and methods for inhibiting nuclear SET domain-containing protein 2 (NSD2). [Background technology]

[0003] Nuclear receptor-binding SET domain protein 2 (NSD2), also known as Wolf-Hirschhorn syndrome candidate 1 (WHSC1) or multiple myeloma SET domain (MMSET), is an epigenetic modifier that is thought to have a driving role in cancer formation. Both NSD2 overexpression and point mutations that increase its catalytic activity are associated with several human cancers. (Coussens et al., J. Biol. Chem. 293, 13750-13654 (2018)).

[0004] NSD2 is dysregulated by the t(4;14)(p16.3;q32.3) translocation in approximately 15% of multiple myeloma (MM) cases. Increased expression of NSD2 in T(4;14)+ MM cell lines is associated with increased levels of H3K36me2 and an inverse decrease in H3K27me3 levels. Such correlations suggest a causal relationship between NSD2 histone methyltransferase (HMT) activity and MM oncogenes.

[0005] High expression of NSD2 protein has been shown in different human cancer types, including bladder, brain, gastrointestinal, lung, liver, ovarian, skin, uterine, breast, prostate, and glioblastoma. (Coussens et al., Ezponda et al. Oncogene, 32:2882-2890 (2013) supra). Notably, NSD2 is one of the most frequently mutated genes in pediatric cancer genomes. The NSD2SET domain variant E1099K was identified in both acute lymphoblastic leukemia tumors and cell lines with increased H3K36me2 lacking the t(4;14) translocation. Sequencing results of >1000 pediatric cancer genomes representing 21 different cancers revealed the E1099K variant in 14% of t(12;21)ETV6-RUNX1, including acute lymphoblastic leukemia (ALL). NSD2 is also one of the most frequently mutated genes found in mantle cell lymphoma tumors, where both E1099K and T1150A variants have been observed. The E1099K variant has also been reported in chronic lymphocytic leukemia (CLL), lung cancer, and gastric cancer. (Coussens et al., supra). In general, upregulation of NSD2 is associated with aggressive tumor behavior and poor prognosis. (Ezponda et al., supra).

[0006] NSD2 is a promising target for cancer therapy, and there remains a need for selective inhibitors of NSD2. Summary of the Invention

[0007] The present invention provides novel compounds that inhibit NSD2, as well as compositions and methods for treating or preventing diseases or conditions mediated by NSD2.

[0008] In one aspect, the present invention provides a compound of formula (I): [ka] or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof, wherein: A is N or CR 9 and R 9 is hydrogen or halo, L is a bond or C 1-4 is alkylene, R 1 is H, or R 1 and R 2 together with NH form a 5-8 membered heterocyclyl containing 1-2 heteroatoms selected from N, O and S as ring members, said 5-8 membered heterocyclyl being unsubstituted or substituted by an oxo substituent; R 2 teeth, (i) Hydrogen, -C 1-6 Alkyl, -HaloC 1-6 Alkyl, -hydroxyC 1-6 Alkylene, -HydroxyhaloC 1-6 Alkylene, -C 1-6 Alkoxy C 1-6 Alkylene, -HaloC 1-6 Alkoxy C 1-6 Alkylene, or -C 3-8 Cycloalkoxy(C 1-6 alkyl), (ii) Cyano, -cyanoC 1-6 Alkylene, -C 1-6 Alkylthio C 1-6 Alkyl, -C 2-6 Alkenyl, -HaloC 2-6 Alkenyl, -C 2-6 Alkynyl, -C 1-4 Alkyl SOC 1-4 Alkyl, -C 1-4 AlkylSO 2 C 1-4 Alkyl, -SO 2 R 8 , or -C(C 1-4 alkyl)=NO(C 1-4 alkyl), (iii)-C 1-4 Alkylcarbonyl, -(CR a R b ) p -C(=O)-OR 10, or -C(=O)-(CR a R b ) q R 11 -(In the formula, R 11 is C 3-7 cycloalkyl, 5- to 6-membered heterocyclyl, or 5- to 6-membered heteroaryl, each of which is independently unsubstituted or C 1-6 Alkyl or C 1-6 substituted with alkoxy; (iv)-(CR a R b ) r -C(=O)-NR 12 R 13 (In the formula, R 12 is hydrogen or C 1-6 is alkyl, R 13 is hydrogen, -C 1-6 alkyl or 5-6 membered heterocycle, or R 12 and R 13 taken together form a 5- to 6-membered heterocycle, said 5- to 6-membered heterocycle being unsubstituted or substituted with alkyl; (v) 5-6-membered heterocyclyl C 0-6 Alkyl or 5-6 membered heterocyclyl (haloC 1-4 alkyl), wherein each said heterocyclyl group is unsubstituted or substituted by oxo; (vi) 5- to 9-membered heteroaryl C 0-6 Alkyl or 5-9 membered heteroaryl (haloC 1-4 alkyl), wherein each said heteroaryl group is unsubstituted or 1-4 Alkyl, -HaloC 1-4 Alkyl, -hydroxyC 1-4 Alkylene, -C 1-4 Alkoxy, -HaloC 1-4 Alkoxy, halo, hydroxy, cyano, oxide, -aminocarbonylC 0-6 Alkyl, -C 1-4 Alkylaminocarbonyl C 0-6 Alkyl, -diC 1-4 Alkylaminocarbonyl C 0-6 Alkyl or -C3-7 substituted by cycloalkyl; R 3a , R 3b , R 4a , R 4b , R 5a , R 5b , R 6a , and R 6b are independently hydrogen, halo, cyano, hydroxyl, -C 1-6 Alkyl, -HaloC 1-6 Alkyl, -hydroxyC 1-6 Alkylene, -C 1-6 Alkoxy, -C 1-6 Alkoxy C 1-6 Alkylene, -HaloC 1-6 Alkoxy C 1-6 Alkylene, -HydroxyhaloC 1-6 Alkylene, aryl, -C(=O)-OR 14 , or -(CR a R b ) s -C(=O)-NR 15 R 16 or R 3a and R 3b , R 4a and R 4b , R 5a and R 5b , or R 6a and R 6b forms an oxo substituent, R 7 -H, -C 1-4 Alkoxy, Halo, C 1-4 alkyl, or 3-8 membered heterocyclyl unsubstituted or substituted by halo; R 8 is C 3-8 Cycloalkyl(C 0-6 alkyl); 4-6 membered heterocyclyl containing 1-3 heteroatoms selected from N, O, and S 0-6 alkyl; aryl or 5-9 membered heteroaryl containing 1-3 heteroatoms selected from N, O, and S; 0-6 is alkyl, R 8is unsubstituted or contains 1 to 3 R 17 is replaced by R 17 is halo, hydroxy, cyano, -C 1-6 Alkyl, -HaloC 1-6 Alkyl, -C 1-6 Alkoxy, -HaloC 1-6 Alkoxy, -NR a C(=O)CR c =C(R c ) 2 , or -(CR a R b ) t -NR a -C(=O)-R 18 and R a , R b , R c , R 10 , R 14 , R 15 , and R 16 are independently hydrogen or -C 1-4 is alkyl, R 18 -C 1-4 Alkyl, or -C 1-4 is haloalkyl, p, q, r, s, and t are independently 0 to 4.

[0009] In another aspect, the present invention provides pharmaceutical compositions comprising a therapeutically effective amount of a compound of formula (I) or a subformula thereof or a pharma- ceutically acceptable salt thereof and one or more pharma- ceutically acceptable carriers.

[0010] In yet another aspect, the present invention provides a combination, in particular a pharmaceutical combination, comprising a therapeutically effective amount of a compound of formula (I) or a subformula thereof, or a pharma- ceutically acceptable salt thereof, and one or more therapeutically active agent(s).

[0011] The compounds of the present invention can be used alone or in combination with one or more therapeutically active agent(s) to treat or prevent diseases or conditions mediated by NSD2, more particularly diseases or conditions characterized by overexpression or unwanted upregulation of NSD2. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] The present invention provides compositions and methods for treating or preventing diseases or conditions mediated by NSD2.

[0013] definition For purposes of interpreting this specification, the following definitions will apply and where appropriate, terms used in the singular will also include the plural and vice versa.

[0014] As used herein, "-C 1-6 The term "alkyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing no unsaturation, having from 1 to 6 carbon atoms, and attached to the remainder of the molecule by a single bond. 1-4 The term "alkyl" should be construed accordingly. 1-6 Examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, and 1,1-dimethylethyl (t-butyl).

[0015] As used herein, "-C 2-6 The term "alkenyl" refers to a straight or branched hydrocarbon chain radical group consisting solely of carbon and hydrogen atoms, containing at least one double bond, having from 2 to 6 carbon atoms and attached to the remainder of the molecule by a single bond. 2-4 The term "alkenyl" should be construed accordingly. 2-6 Examples of alkenyl include, but are not limited to, ethenyl, prop-1-enyl, but-1-enyl, pent-1-enyl, pent-4-enyl, and penta-1,4-dienyl.

[0016] As used herein, "-C 2-6 The term "alkynyl" refers to a straight or branched hydrocarbon chain radical group consisting solely of carbon and hydrogen atoms, containing at least one triple bond, having from 2 to 6 carbon atoms and attached to the remainder of the molecule by a single bond. 2-4 The term "alkynyl" should be construed accordingly. 2-6 Examples of alkynyl include, but are not limited to, ethynyl, prop-1-ynyl, but-1-ynyl, pent-1-ynyl, pent-4-ynyl, and penta-1,4-diynyl.

[0017] As used herein, "-C 1-6 The term "alkoxy" refers to a group of the formula -OR a In the formula, R a In general, C 1-6 It is an alkyl group. 1-6 Examples of alkoxy include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, pentoxy, and hexoxy.

[0018] As used herein, "-C 1-6 Alkoxy C 1-6 The term "alkylene" refers to a group of the formula -R a -OR a In the formula, each R a are independently C as defined above 1-6 are alkyl groups. The oxygen atom may be bonded to any carbon atom in any alkyl group. 1-6 Alkoxy C 1-6 Examples of alkyl include, but are not limited to, methoxy-methyl, methoxy-ethyl, ethoxy-ethyl, 1-ethoxy-propyl, and 2-methoxy-butyl.

[0019] As used herein, "-C 1-4 The term "alkylcarbonyl" refers to a group of the formula -C(=O)-R aIn the formula, R a is the C defined above. 1-4 It is an alkyl group.

[0020] As used herein, "-C 1-4 Alkylthio C 1-4 The term "alkyl" means a group of the formula -R a -SR a In the formula, each R a are independently C as defined above 1-4 It is an alkyl group.

[0021] As used herein, "-hydroxyC 1-6 The term "alkylene" means 1-6 C, as defined above, in which one of the hydrogen atoms of the alkyl group is replaced by OH. 1-6 It means an alkyl group. Hydroxy C 1-6 Examples of alkyl include, but are not limited to, ethane-1-olyl, 2-methylpropan-1-olyl, hydroxy-methyl, 2-hydroxy-ethyl, 2-hydroxy-propyl, 3-hydroxy-propyl, and 5-hydroxy-pentyl.

[0022] As used herein, "-aminocarbonyl C 0-6 The term "alkyl" means a group of the formula -R a -C(=O)-NH 2 In the formula, R a is a single bond or C as defined above 1-6 It is an alkyl group.

[0023] As used herein, "-C 1-4 Alkylaminocarbonyl C 0-6 The term "alkyl" means a group of the formula -R a1 -C(=O)-NH-R a2 In the formula, R a1 is a single bond or C as defined above 1-6 is an alkyl group, R a2is C as defined above. 1-6 (which is an alkyl group).

[0024] As used herein, "-diC 1-4 Alkylaminocarbonyl C 0-6 The term "alkyl" refers to a group of the formula -R a1 -C(=O)-N(R a2 )-R a2 In the formula, R a1 is a single bond or C as defined above 1-6 is an alkyl group, and each R a2 is the C defined above. 1-4 are alkyl groups which may be the same or different.

[0025] As used herein, "-C 3-8 Cycloalkyl C 0-6 The term "alkyl" refers to a group consisting solely of carbon and hydrogen atoms, having 3 to 8 carbon atoms, and containing a single bond (i.e., C 3-8 cycloalkyl) or C as defined above 1-6 It refers to a stable monocyclic saturated hydrocarbon group that is attached to the rest of the molecule by an alkyl group. 3-7 Cycloalkyl" and "C 3-7 Cycloalkyl C 0-4 The term "alkyl" should be construed accordingly. 3-8 Cycloalkyl C 0-6 Examples of alkyl include, but are not limited to, cyclopropyl, cyclopropyl-methyl, cyclobutyl, cyclobutyl-ethyl, cyclopentyl, cyclopentyl-propyl, cyclohexyl, cycloheptaptyl, and cyclooctyl.

[0026] As used herein, "-C 3-8 Cycloalkoxy C 1-6 The term "alkyl" refers to a group of the formula -R a -OR b In the formula, R a are independently C as defined above 1-6 is an alkyl group, R b is the C defined above.3-8 Cycloalkyl. C 3-8 Cycloalkoxy C 1-6 Examples of alkyl include, but are not limited to, cyclopropoxymethyl and cyclobutoxymethyl.

[0027] "Halo" refers to bromo, chloro, fluoro, or iodo.

[0028] As used herein, "-haloC 1-6 The term "alkyl" refers to a C alkyl group, as defined above, substituted by one or more halo groups, as defined above. 1-6 Refers to an alkyl group. HaloC 1-6 Examples of alkyl include, but are not limited to, trifluoromethyl, difluoromethyl, fluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,3-dibromopropan-2-yl, 3-bromo-2-fluoropropyl, and 1,4,4-trifluorobutan-2-yl. 1-4 The term "alkyl" should be construed accordingly.

[0029] As used herein, "-haloC 1-6 Alkoxy C 1-4 The term "alkylene" refers to a group of the formula -R a1 -OR a2 In the formula, R a1 is the C defined above. 1-6 is an alkyl group, R a2 is the halo C defined above 1-6 Alkyl. HaloC 1-6 Alkoxy C 1-4 Examples of alkyl include, but are not limited to, (difluoromethoxyl)methyl, (2,2,2-trifluoroethoxy)methyl, and (2,2-difluoroethoxy)methyl.

[0030] As used herein, "-hydroxyl haloC 1-6 The term "alkylene" refers to a haloC, as defined above, substituted with one or more hydroxyl groups. 1-6Refers to the alkyl group. Hydroxyl halo C 1-6 Examples of alkyl include, but are not limited to, 2,2-difluoroethan-1-olyl, 2-fluoroethan-1-olyl, and 2,2,2-trifluoroethan-1-olyl.

[0031] As used herein, "-cyanoC 1-6 The term "alkylene" refers to a group of the formula -R a —CN group, where R a is the C defined above. 1-4 It is an alkyl group.

[0032] As used herein, "-haloC 2-6 The term "alkenyl" refers to any of the C1, C2, and C3 groups defined above substituted by one or more halo groups defined above. 2-6 Refers to an alkenyl group.

[0033] As used herein, the term "heterocyclyl" or "heterocyclic" refers to a stable 4-7 membered non-aromatic monocyclic group containing one, two, or three heteroatoms independently selected from nitrogen, oxygen, and sulfur. The heterocyclyl group may be attached via a carbon atom or a heteroatom. The term "5- to 6-membered heterocyclyl" shall be interpreted accordingly. Examples of heterocyclyl include, but are not limited to, azetidinyl, oxetanyl, pyrrolinyl, pyrrolidyl, tetrahydrofuryl, tetrahydrothienyl, piperidyl, piperazinyl, tetrahydropyranyl, or morpholinyl or perhydroazepinyl.

[0034] As used herein, "heterocyclyl C 0-6 The term "alkyl" means a C alkyl group defined above, 1-6 It refers to a heterocycle as defined above attached to the remainder of the molecule by an alkyl group.

[0035] As used herein, the term "heteroaryl" refers to a 5-9 membered aromatic monocyclic or fused ring group containing 1, 2, 3, or 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur. The heteroaryl group may be attached via a carbon atom or a heteroatom. The term "5-6 membered heteroaryl" shall be interpreted accordingly. Examples of 5-6 membered monocyclic heteroaryl include, but are not limited to, furyl, pyrrolyl, thienyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl, tetrazolyl, pyrazinyl, pyridazinyl, pyrimidyl, or pyridyl. Examples of fused heteroaryls include, but are not limited to, 9-membered heteroaryls, such as benzofuranyl, 2,3-dihydrobenzofuranyl, 1,3-dihydroisobenzofuranyl, benzo[d][1,3]dioxol-5-yl, imidazo[1,2-a]pyridinyl, pyrazolo[1,5-a]pyridinyl, 1H-indazolyl, and 1H-benzo[d]-imidazolyl.

[0036] As used herein, "heteroaryl C 0-6 The term "alkyl" means a C alkyl group defined above, 1-6 It refers to a heteroaryl ring as defined above attached to the remainder of the molecule by an alkyl group.

[0037] As used herein, "heteroaryl (haloC 1-4 The term "halo (alkyl)" refers to haloC as defined above. 1-4 Heteroaryl (haloC) refers to a heteroaryl ring as defined above that is bonded to the remainder of the molecule by an alkyl. 1-4 An illustrative example of alkyl) is fluoro(pyridin-2-yl)methyl.

[0038] As used herein, "IC 50 " refers to the molar concentration of inhibitor or modulator that produces 50% inhibition.

[0039] As used herein, "protected derivatives" refers to derivatives of inhibitors in which reactive sites are blocked with protecting groups. Protected derivatives may be useful in the preparation of inhibitors or may be active as inhibitors themselves. Examples of protecting groups include, but are not limited to, acetyl, tetrahydropyran, methoxymethyl ether, β-methoxyethoxymethyl ether, ρ-methoxybenzyl, methylthiomethyl ether, pivaloyl, silyl ether, carbobenzyloxy, benzyl, tert-butoxycarbonyl, ρ-methoxyphenyl, 9-fluorenylmethyloxycarbonyl, acetal, ketal, acylal, dithiane, methyl ester, benzyl ester, tert-butyl ester, and silyl ester. A comprehensive list of suitable protecting groups can be found in TW Greene, Protecting Groups in Organic Synthesis, 3rd edition, John Wiley & Sons, Inc. 1999.

[0040] The term "disease or condition mediated by nuclear SET domain-containing protein 2 (NSD2)" refers to a disease or condition that is directly or indirectly regulated by NSD2.

[0041] As used herein, the term "subject" refers to mammals, primates (e.g., humans, male or female), dogs, rabbits, guinea pigs, pigs, rats, and mice. In certain embodiments, the subject is a primate. In yet other embodiments, the subject is a human.

[0042] As used herein, the terms "inhibit," "inhibition," or "inhibiting" refer to the reduction or suppression of a given condition, symptom, or disorder, or disease, or a significant decrease in the baseline activity of a biological activity or process.

[0043] As used herein, the terms "treat," "treating," or "treatment" mean to alleviate or ameliorate a disease or disorder, including those that may not be discernible to the patient (i.e., to delay or arrest the onset of the disease or at least one of its clinical symptoms), or to alleviate or ameliorate at least one physical parameter or biomarker associated with a disease or disorder.

[0044] As used herein, the terms "prevent," "preventing," or "prevention" of any disease or disorder means the prophylactic treatment of a disease or disorder or the delay of the onset or progression of a disease or disorder.

[0045] As used herein, a subject is "in need of" a treatment if such subject would benefit biologically, medically, or in quality of life from the treatment.

[0046] As used herein, the term "therapeutically effective amount" of a compound of the invention refers to an amount of a compound of the invention that elicits a biological or medical response in a subject, such as reducing or inhibiting enzyme or protein activity, ameliorating symptoms, alleviating a condition, slowing or delaying the progression of a disease, or preventing a disease.

[0047] As used herein, the term "anti-cancer agent" or antineoplastic agent refers to a therapeutic agent useful for treating or controlling the growth of cancer cells.

[0048] As used herein, the term "anti-inflammatory agent" refers to a therapeutic agent that reduces inflammation (redness, swelling, and / or pain) in the body. Anti-inflammatory agents block certain substances in the body that cause inflammation.

[0049] As used herein, the term "pharmaceutical composition" refers to a compound of the present invention or a pharma- ceutical acceptable salt thereof, together with at least one pharma- ceutical acceptable carrier, in a form suitable for oral or parenteral administration.

[0050] As used herein, the term "pharmaceutically acceptable carrier" refers to a substance useful for the preparation or use of a pharmaceutical composition, and is known to those skilled in the art (e.g., Remington The Science and Practice of Pharmacy, 22 nd Ed. Pharmaceutical Press, 2013, pp. 1049-1070), suitable diluents, solvents, dispersion media, surfactants, antioxidants, preservatives, isotonicity agents, buffers, emulsifiers, absorption delaying agents, salts, drug stabilizers, binders, excipients, disintegrants, lubricants, wetting agents, sweeteners, flavoring agents, dyes, and combinations thereof.

[0051] As used herein, the terms "a," "an," "the," and similar terms as used in the context of the present invention (particularly in the context of the claims) are to be construed to encompass both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context.

[0052] Description of the Preferred Embodiments The present invention provides novel compounds that inhibit NSD2, as well as compositions and methods for treating or preventing conditions mediated by NSD2.

[0053] Various enumerated embodiments of the invention are described herein. The features specified in each embodiment may be combined with other specified features to provide further embodiments of the invention.

[0054] Embodiment 1. A compound of formula (I) above, or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof.

[0055] The compound of formula (I) according to Embodiment 1, wherein A is N, or an enantiomer, enantiomer mixture, or pharmaceutically acceptable salt thereof.

[0056] Embodiment 3. R 3a is hydrogen or halo, and R 3b is hydrogen, halo, -hydroxyl, -C 1-6 alkoxy, or cyano, or R 4a is hydrogen or halo, and R 4b is hydrogen, halo, -C 1-6 alkoxyC 1-6 alkylene, -C 1-6 alkyl, or -haloC 1-6 alkyl, or R 5a is hydrogen, and R 5b is hydrogen or -C 1-6 alkyl, or R 5a and R 5b together form an oxo substituent, the compound of formula (I) according to Embodiment 1 or 2, or an enantiomer, enantiomer mixture, or pharmaceutically acceptable salt thereof.

[0057] Embodiment 4. R 6a is hydrogen or halo, R 6b is hydrogen, -haloC 1-6 alkoxyC 1-6 alkylene, -hydroxyC 1-6 alkylene, -hydroxyhaloC 1-6 alkylene, carboxyl, phenyl, or -(CR a R b ) t -C(O)-NR 15 R 16 is, R a , R b , R 15 and R 16 are independently hydrogen or -C1-4 is alkyl, t is between 0 and 1, or R 6a and R 6b A compound of formula (I) as defined in any one of embodiments 1 to 3, or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof, wherein together form an oxo substituent.

[0058] Embodiment 5.R 7 But, H, -C 1-4 The compound of formula (I) according to any one of embodiments 1 to 4, or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof, wherein R is alkoxy, or halo.

[0059] Embodiment 6.R 1 , R 3a , R 3b , R 4a , R 4b , R 5a , R 5b , R 6a , R 6b , and R 7 A compound of formula (I) according to any one of embodiments 1 to 5, or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof, wherein is hydrogen.

[0060] Embodiment 7. The compound of formula (I) according to embodiment 1, wherein the compound is a compound of formula (II). [ka] or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof.

[0061] Embodiment 8. The compound is a compound of formula (IIA), a compound of formula (II) as described in embodiment 7, [ka] or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof.

[0062] Embodiment 9.R 2 is hydrogen, -C 1-6 Alkyl, -HaloC 1-6 Alkyl, -hydroxyC 1-6 Alkylene, -HydroxyhaloC 1-6 Alkylene, -C 1-6 Alkoxy C 1-6 Alkylene, -HaloC 1-6 Alkoxy C 1-6 Alkylene, or -C 3-8 Cycloalkoxy(C 1-6 alkyl), or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof.

[0063] Embodiment 10.R 2 But -C 1-4 Alkyl carbonyl, -(CR a R b ) p -C(O)-OR 10 (In the formula, R 10 is hydrogen or -C 1-4 alkyl), -C(O)-(CR a R b ) q -R 11 (In the formula, R 11 But, C 3-6 cycloalkyl, 5- to 6-membered heterocyclyl, or 5- to 6-membered heteroaryl, each of which is independently unsubstituted or 1-6 Alkyl or -C 1-6 alkoxy), or -(CR a R b ) r -C(O)-NR 12 R 13 (In the formula, R 12 is hydrogen or -C 1-6 is alkyl, R 13 is hydrogen, -C 1-6 alkyl, or a 5- to 6-membered heterocycle; or R 12 and R 13 together form a 5- to 6-membered heterocyclic ring, the 5- to 6-membered heterocyclic ring being unsubstituted or 1-4 substituted with alkyl; R a , R b , and R 11 are independently hydrogen or -C 1-4 is alkyl, The compound according to any one of embodiments 1 to 8, wherein p, q and r are independently 0 to 2, or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof.

[0064] Embodiment 11.R 2 but, pyridin-2-yl, (pyridin-2-yl)methyl, or (pyridin-2-yl)ethyl, wherein the pyridin-2-yl is independently unsubstituted or contains one to two -C 1-4 Alkyl, -C 1-4 Haloalkyl, hydroxyl, -Hydroxy C 1-4 Alkylene, Cyano, -C 1-4 Alkoxy, -C 1-6 Alkoxy C 1-6 Alkylene, -HaloC 1-4 Alkoxy, halo or -C 3-7 substituted with cycloalkyl; Thiazol-4-yl, thiazol-2-ylmethyl, oxazol-2-yl, oxazol-2-ylmethyl, pyridazin-3-yl, pyridazin-3-ylmethyl, pyrazin-2-yl, pyrazin-2-ylmethyl, pyrimidin-4-yl, pyrimidin-4-ylmethyl, each of which is independently unsubstituted or -C 1-4 substituted with alkyl or halo; 1H-pyrazol-3-yl, (1H-pyrazol-3-yl)methyl, 1-methyl-1H-pyrazol-4-yl, (1,2,3-triazol-4-yl), isoxazol-3-yl, each of which is independently unsubstituted or -C1-4 Alkyl, halo or -C 3-7 cycloalkyl substituted), 1,4-dioxan-2-yl, Pyridin-2(1H)-onyl, and The compound according to any one of embodiments 1 to 8, selected from pyridin-1-oxid-2-yl, or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof.

[0065] Embodiment 12.R 8 The compound according to any one of embodiments 1-11, or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof, wherein is cyclopropyl, cyclobutyl, cyclohexyl, or azetidinyl.

[0066] Embodiment 13.R 8 is 2,3-dihydrobenzofuran-5-yl, 1,3-dihydroisobenzofuran-5-yl, benzo[d][1,3]dioxol-5-yl, imidazo[1,2-a]pyridin-6-yl, or pyrazolo[1,5-a]pyridin-6-yl, or an enantiomer, mixture of enantiomers, or a pharma- ceutically acceptable salt thereof.

[0067] Embodiment 14.R 8 is unsubstituted or contains 1 to 2 halo, -C 1-6 Alkyl or -C 1-6 alkoxy substituted, pyridyl; unsubstituted or substituted with 1-2 halo or -C 1-6 1H-indazol-5-yl, substituted with alkyl; or unsubstituted or -C 1-6 The compound according to any one of embodiments 1 to 13, which is 1H-benzo[d]-imidazol-6-yl substituted with alkyl, or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof.

[0068] Embodiment 15.R 8However, 1 to 3 R 17 is a phenyl substituted with R 17 But halo, hydroxy, cyano, -C 1-6 Alkyl, -HaloC 1-6 Alkyl, -C 1-6 Alkoxy, -HaloC 1-6 Alkoxy, -NR d C(O)CR d =C(R d ) 2 , or -(CR a R b ) t -NR d -C(O)-R 18 and R a , R b , and R d are independently hydrogen or -C 1-4 is alkyl, R 18 But -C 1-4 is haloalkyl, The compound according to any one of embodiments 1 to 13, or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof, wherein t is 0 to 1.

[0069] Embodiment 16.R 8 However, 1 to 3 R 17 phenyl substituted with R 17 But halo, hydroxy, -C 1-6 Alkyl, -HaloC 1-6 Alkyl, -C 1-6 Alkoxy, or -HaroC 1-6 The compound according to any one of embodiments 1 to 13, or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof, wherein:

[0070] Embodiment 17. A compound of formula (II): [ka] or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof. (In the formula, R 2 is hydrogen, -C 1-6 Alkyl, -HaloC 1-6 Alkyl, -hydroxyC 1-6 Alkylene, -HydroxyhaloC 1-6 Alkylene, -C 1-6 Alkoxy C 1-6 Alkylene, -HaloC 1-6 Alkoxy C 1-6 Alkylene, or -C 3-8 Cycloalkoxy(C 1-6 alkyl), R 8 1 to 3 R 17 is a phenyl substituted with R 17 ,Halo,-C 1-6 Alkyl, -HaloC 1-6 Alkyl, -C 1-6 Alkoxy or -haloC 1-6 (alkoxy).

[0071] Embodiment 18. The compound is a compound of formula (IIA), a compound of formula (II) as described in embodiment 17, [ka] or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof.

[0072] Embodiment 19.R 2 is hydrogen, 2,2-difluoroethyl, 2-methylpropan-1-olyl, ethane-1-olyl, 2,2-difluoroethane-1-olyl, 2-fluoroethane-1-olyl, 2,2,2-trifluoroethane-1-olyl, difluoromethoxyl, or 2,2,2-trifluoroethoxyl, or an enantiomer, mixture of enantiomers, or a pharma-ceutically acceptable salt thereof.

[0073] Embodiment 20.R 2 but, [ka] 19. The compound according to any one of embodiments 1 to 18, wherein:

[0074] Embodiment 21. The compound of any one of embodiments 1-20, or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof, wherein the compound is a compound in Table 2.

[0075] Embodiment 22. The compound is 1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(2,4,5-trifluorophenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol, 1-(3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(2-(difluoromethyl)-4-fluorophenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol; 1-(3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(2,5-difluoro-4-methoxyphenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol, 1-(3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(2-(difluoromethyl)-4-methoxyphenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol; 1-(3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(3,5-difluoro-4-methoxyphenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol, 1-(3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(4-methoxy-2-(trifluoromethyl)phenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol, 1-(3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(2-chloro-4-methoxyphenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol, 1-(3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(2-(difluoromethyl)-4,5-difluorophenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol; 1-(3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(2,3,4-trifluorophenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol, 1-(3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(4-fluoro-2-methoxyphenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol, and 22. The compound according to embodiment 21, selected from the group consisting of: 1-(3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6'-chloro-5'-fluoro-[2,2'-bipyridine]-5-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol; or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof.

[0076] Embodiment 23. The compound of embodiment 22, wherein the compound is in the (R) configuration, the (S) configuration, or a mixture thereof.

[0077] Embodiment 24. The compound of embodiment 1, wherein the compound is (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(2,4,5-trifluorophenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol, or a pharmaceutically acceptable salt thereof. Alternatively, the compound is (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(2,4,5-trifluorophenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol, or a pharmaceutically acceptable salt thereof.

[0078] Embodiment 25. The compound of embodiment 1, wherein the compound is (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(2-(difluoromethyl)-4-fluorophenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol, or a pharma-ceutically acceptable salt thereof. Alternatively, the compound is (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(2-(difluoromethyl)-4-fluorophenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol, or a pharma-ceutically acceptable salt thereof.

[0079] Embodiment 26. The compound of embodiment 1, wherein the compound is (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(2,5-difluoro-4-methoxyphenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol, or a pharmaceutically acceptable salt thereof. Alternatively, the compound is (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(2,5-difluoro-4-methoxyphenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol, or a pharmaceutically acceptable salt thereof.

[0080] Embodiment 27. The compound of embodiment 1, wherein the compound is (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(2-(difluoromethyl)-4-methoxyphenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol, or a pharma-ceutically acceptable salt thereof. Alternatively, the compound is (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(2-(difluoromethyl)-4-methoxyphenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol, or a pharma-ceutically acceptable salt thereof.

[0081] Embodiment 28. The compound of embodiment 1, wherein the compound is (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(3,5-difluoro-4-methoxyphenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol, or a pharma-ceutically acceptable salt thereof. Alternatively, the compound is (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(3,5-difluoro-4-methoxyphenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol, or a pharma-ceutically acceptable salt thereof.

[0082] Embodiment 29. The compound of embodiment 1, wherein the compound is (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(4-methoxy-2-(trifluoromethyl)phenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol, or a pharma-ceutically acceptable salt thereof. Alternatively, the compound is (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(4-methoxy-2-(trifluoromethyl)phenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol, or a pharma-ceutically acceptable salt thereof.

[0083] Embodiment 30. The compound of embodiment 1, wherein the compound is (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(2-chloro-4-methoxyphenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol, or a pharma-ceutically acceptable salt thereof. Alternatively, the compound is (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(2-chloro-4-methoxyphenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol, or a pharma-ceutically acceptable salt thereof.

[0084] Embodiment 31. The compound of embodiment 1, wherein the compound is (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(2-(difluoromethyl)-4,5-difluorophenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol, or a pharma-ceutically acceptable salt thereof. Alternatively, the compound is (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(2-(difluoromethyl)-4,5-difluorophenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol, or a pharma-ceutically acceptable salt thereof.

[0085] Embodiment 32. The compound of embodiment 1, wherein the compound is (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(2,3,4-trifluorophenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol, or a pharma-ceutically acceptable salt thereof. Alternatively, the compound is (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(2,3,4-trifluorophenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol, or a pharma-ceutically acceptable salt thereof.

[0086] Embodiment 33. The compound of embodiment 1, wherein the compound is (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(4-fluoro-2-methoxyphenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol, or a pharma-ceutically acceptable salt thereof. Alternatively, the compound is (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(4-fluoro-2-methoxyphenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol, or a pharma-ceutically acceptable salt thereof.

[0087] Embodiment 34. The compound of embodiment 1, wherein the compound is (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6'-chloro-5'-fluoro-[2,2'-bipyridine]-5-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol, or a pharmaceutically acceptable salt thereof. Alternatively, the compound is (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6'-chloro-5'-fluoro-[2,2'-bipyridine]-5-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol, or a pharmaceutically acceptable salt thereof.

[0088] Embodiment 35. A pharmaceutical composition comprising a compound according to any one of embodiments 1 to 34 and one or more pharma- ceutically acceptable carriers.

[0089] Embodiment 36. A combination comprising a compound according to any one of embodiments 1 to 34 and one or more additional therapeutically active agents.

[0090] Embodiment 37. The combination of embodiment 36, wherein said one or more additional therapeutically active agents is an anti-cancer agent, an analgesic agent, an anti-inflammatory agent, or a combination thereof.

[0091] Embodiment 38. A compound according to any one of embodiments 1 to 34, optionally in combination with a second therapeutic agent, for use in the treatment of a disease or condition mediated by nuclear SET domain-containing protein 2 (NSD)2.

[0092] Embodiment 39. The compound of embodiment 38, wherein the second therapeutic agent is an anti-cancer agent, an analgesic agent, an anti-inflammatory agent, or a combination thereof.

[0093] Embodiment 40. Use of a compound according to any one of embodiments 1 to 34, optionally in combination with a second therapeutic agent, in the manufacture of a medicament for a disease or condition mediated by NSD2.

[0094] Embodiment 41. A method for treating a disease or condition mediated by nuclear SET domain-containing protein 2 (NSD2), comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to any one of embodiments 1 to 34, optionally in combination with a second therapeutic agent, thereby treating said disease or condition mediated by NSD2.

[0095] Embodiment 42. A method for treating a disease or condition that would benefit from or be treatable by inhibition of nuclear SET domain-containing protein 2 (NSD2), comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to any one of embodiments 1 to 34, optionally in combination with a second therapeutic agent, thereby treating the disease or condition that would benefit from or be treatable by inhibition by NSD2.

[0096] Embodiment 43. The method of embodiment 41 or 42, wherein the disease or condition mediated by NSD2 or the disease or condition that benefits from or is treatable by inhibition of NSD2 is breast cancer, cervical cancer, skin cancer (particularly squamous cell carcinoma), ovarian cancer, gastric cancer, prostate cancer, pancreatic cancer, lung cancer, hepatocellular carcinoma, head and neck cancer, peripheral nerve sheath tumor, osteosarcoma, multiple myeloma, neuroblastoma, leukemia (particularly acute lymphoblastic leukemia), non-Hodgkin's lymphoma (particularly mantle cell lymphoma), or pulmonary arterial hypertension.

[0097] Embodiment 44. The method of embodiment 41 or 42, wherein the disease or condition mediated by NSD2 or that benefits from or is treatable by inhibition of NSD2 is lung cancer.

[0098] Embodiment 45. The method of embodiment 44, wherein the lung cancer is small cell or non-small cell lung cancer.

[0099] Embodiment 46 The method according to embodiment 41 or 42, wherein the disease or condition mediated by NSD2 or that benefits from or is treatable by inhibition of NSD2 is leukemia.

[0100] Embodiment 47. The method of embodiment 46, wherein the leukemia is acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic myelogenous leukemia (CML), or chronic myelomonocytic leukemia (CMML).

[0101] Embodiment 48. The method of embodiment 41 or 42, wherein the disease or condition mediated by NSD2 or that benefits from or is treatable by inhibition of NSD2 is skin cancer.

[0102] Embodiment 49. The method of embodiment 48, wherein the skin cancer is melanoma, basal cell carcinoma, or squamous cell carcinoma.

[0103] Embodiment 50. The method of embodiment 41 or 42, wherein the disease or condition mediated by NSD2 or that benefits from or is treatable by inhibition of NSD2 is lymphoma.

[0104] Embodiment 51. The method of embodiment 50, wherein the lymphoma is Hodgkin's lymphoma or non-Hodgkin's lymphoma.

[0105] Embodiment 52. The method of embodiment 50, wherein the lymphoma is mantle cell lymphoma or diffuse large B-cell lymphoma.

[0106] Embodiment 53. The method of embodiment 41 or 42, wherein the disease or condition mediated by NSD2 or that benefits from or is treatable by inhibition of NSD2 is myeloma.

[0107] Embodiment 54 The method of any one of embodiments 41-53, wherein the compound is administered orally.

[0108] Unless otherwise indicated, the term "compounds of the invention" or "compound of the invention" refers to compounds of formula (I), subformulas thereof, and exemplified compounds, as well as salts thereof, and all stereoisomers (including diastereoisomers and enantiomers), rotamers, tautomers, and isotopically labeled compounds (including deuterium substitutions), and inherently formed moieties.

[0109] Depending on the selection of starting materials and procedures, the compounds may exist in one of the possible stereoisomers or as a mixture thereof, depending on the number of asymmetric carbon atoms, e.g., as pure optical isomers or as stereoisomeric mixtures, such as racemic and diastereoisomeric mixtures. The present invention is intended to include all such possible stereoisomers, including racemic mixtures, diastereomeric mixtures, and optically pure forms. Optically active (R) and (S) stereoisomers may be prepared using chiral synthons or chiral reagents or resolved using conventional techniques. Where possible, substituents at atoms with unsaturated double bonds may exist in cis-(Z) or trans-(E) form. When the compound contains a disubstituted cycloalkyl, the cycloalkyl substituent may have a cis or trans configuration.

[0110] Any asymmetric atom (e.g., carbon, etc.) of the compound(s) of the invention can be present in racemic or enantiomerically enriched, e.g., (R), (S), or (R,S) configuration. In certain embodiments, each asymmetric atom has at least 50% enantiomeric excess, at least 60% enantiomeric excess, at least 70% enantiomeric excess, at least 80% enantiomeric excess, at least 90% enantiomeric excess, at least 95% enantiomeric excess, or at least 99% enantiomeric excess in the (R) or (S) configuration.

[0111] Thus, as used herein, the compounds of the invention may be in the form of one of the possible stereoisomers, rotamers, atropisomers, tautomers, or mixtures thereof, for example, as substantially pure geometric (cis or trans) stereoisomers, diastereomers, optical isomers (enantiomers), racemates, or mixtures thereof.

[0112] Any mixtures of stereoisomers obtained can be separated on the basis of the physical chemical differences of the components, for example, by chromatography and / or fractional crystallization, into pure or substantially pure geometric or optical isomers, diastereomers, racemates.

[0113] Any resulting racemic forms of the compounds of the present invention or intermediates thereof can be resolved into their optical antipodes by known methods, for example by separation of their diastereomeric salts obtained with optically active acids or bases and liberation of the optically active acidic or basic compounds. Thus, in particular, basic moieties can be used to resolve the compounds of the present invention into their optical antipodes, for example by fractional crystallization of salts formed with optically active acids, for example tartaric acid, dibenzoyltartaric acid, diacetyltartaric acid, di-O,O'-p-toluoyltartaric acid, mandelic acid, malic acid, or camphor-10-sulfonic acid. The racemic compounds or racemic intermediates of the present invention can also be resolved by chiral chromatography, for example by high pressure liquid chromatography (HPLC) using a chiral adsorbent.

[0114] Any formula given herein is also intended to represent unlabeled and isotopically labeled forms of compounds.Isotopically labeled compounds have the structure shown by the formula given herein, except that one or more atoms are replaced by atoms with selected atomic mass or mass number.The isotopes that can be incorporated in the compounds of the present invention include, for example, hydrogen isotopes.

[0115] In addition, certain isotopes, particularly deuterium (i.e. 2Incorporation of H or D) may result in certain therapeutic benefits due to greater metabolic stability, such as increased half-life or reduced dosage requirements in vivo, or improved therapeutic index or tolerability. It is understood that deuterium in this context is considered a substituent of the compound of formula (I) or its subformulas. The concentration of deuterium can be defined by the isotopic enrichment factor. The term "isotopic enrichment factor" as used herein refers to the ratio between the isotopic abundance and the natural abundance of a particular isotope. When a substituent in a compound of the present invention is designated as deuterium, such compound has an isotopic enrichment factor for each designated deuterium atom of at least 3500 (52.5% deuterium incorporation at each designated deuterium atom), at least 4000 (60% deuterium incorporation), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium incorporation), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% deuterium incorporation), at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), at least 6600 (99% deuterium incorporation), or at least 6633.3 (99.5% deuterium incorporation). It is understood that the term "isotopic enrichment factor" can be applied to any isotope in the same manner as described for deuterium.

[0116] Other examples of isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, e.g., 3 H, 11 C. 13 C. 14 C. 15 N, 18 F, 31 P, 32 P, 35 S, 36 Cl, 123 I, 124 I, 125 Thus, the present invention includes, for example, 3 H and 14 Radioisotopes such as C, or 2 H and13 It should be understood to include compounds incorporating any one or more of the above isotopes, including those in which non-radioactive isotopes such as C are present. Such isotopically labeled compounds may be useful in metabolic studies ( 14 C), reaction kinetic studies (e.g. 2 H or 3 H), detection or imaging techniques such as positron emission tomography (PET) or single photon emission computed tomography (SPECT), including drug or substrate tissue distribution assays, or radiation treatment of patients. 18 F or labeled compounds may be particularly desirable for PET or SPECT studies. Isotopically labeled compounds of formula (I) or subformulas thereof can generally be prepared by conventional techniques known to those skilled in the art, or by processes similar to those described in the accompanying examples, substituting the appropriate isotopically labeled reagent for the non-labeled reagent previously used.

[0117] The compounds of the present invention may be obtained in free form or as a salt thereof. As used herein, the term "salt" or "salts" refers to an acid addition salt or a base addition salt of the compounds of the present invention. "Salt" specifically includes "pharmaceutically acceptable salts". The term "pharmaceutically acceptable salts" refers to salts that retain the biological effectiveness and properties of the compounds of the present invention and are typically not biologically or otherwise undesirable. In many cases, the compounds of the present invention are capable of forming acid and / or base salts by virtue of the presence of amino and / or carboxyl groups or groups similar thereto.

[0118] Pharmaceutically acceptable acid addition salts can be formed with inorganic and organic acids. Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc. Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, sulfosalicylic acid, etc.

[0119] Pharmaceutically acceptable base addition salts can be formed with inorganic and organic bases. Inorganic bases from which salts can be derived include, for example, ammonium salts and metals from columns I-XII of the periodic table. In certain embodiments, salts are derived from sodium, potassium, ammonium, calcium, magnesium, iron, silver, zinc, and copper, with particularly suitable salts including ammonium, potassium, sodium, calcium, and magnesium salts. Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like. Particular organic amines include isopropylamine, benzathine, cholate, diethanolamine, diethylamine, lysine, meglumine, piperazine, and tromethamine.

[0120] In another aspect, the present invention provides an ester of an amine salt, selected from the group consisting of acetate, ascorbate, adipate, aspartate, benzoate, besylate, bromide / hydrobromide, bicarbonate / carbonate, bisulfate / sulfate, camphorsulfonate, caprate, chloride / hydrochloride, chlorotheophylline, citrate, ethanedisulfonate, fumarate, gluceptate, gluconate, glucuronate, glutamate, glutarate, glycolate, hippurate, hydroiodide / iodide, isethionate, lactate, lactobionate, lauryl sulfate, malate, maleate, and the like. The compounds of the invention are provided in the form of an iodate, malonate, mandelate, mesylate, methylsulfate, mucate, naphthate, napsylate, nicotinate, nitrate, octadecanoate, oleate, oxalate, palmitate, pamoate, phosphate / hydrogen phosphate / dihydrogen phosphate, polygalacturonate, propionate, sebacate, stearate, succinate, sulfosalicylate, sulfate, tartrate, tosylate, trifenatate, trifluoroacetate, or xinafoate salt.

[0121] Processes for Making the Compounds of the Invention All methods described herein can be performed in any suitable order unless otherwise indicated or otherwise clearly contradicted by context.

[0122] Compounds of formula (I) may generally be prepared as illustrated in Scheme 1, 1 , R 2 , R 3 , ... are as defined above. As shown in Scheme 1, Boc-protected 3-CN or 3-CO 2 Et piperidine is reacted with a strong base (e.g., NaHMDS) and R 2 In the presence of electrophiles including S N 2 or S N Upon Ar reaction, compounds 2 or 2' bearing a quaternary center were generated, which were further hydrolyzed to the corresponding carboxylic acid 3 or primary amide 3' under either basic (e.g., LiOH) or oxidative hydrolysis (e.g., H2O2, NaOH) conditions. The acid 3 or amide 3' further undergoes Curtius or Hofmann rearrangement to generate the protected quaternary amine 4. After selective Boc deprotection, piperidine 5 reacts with fluorinated pyridine 6 under heating in the presence of an organic base (e.g., DIPEA) to give the coupling product 7. Ester 7 is further reduced by a mild reducing agent (e.g., NaBH4, LiCl) to benzylic alcohol 8, which undergoes Mitsunobu reaction with Boc-protected adenine intermediate B to generate compound 9. After deprotection under acidic or hydrogenation conditions, the chiral target molecule 10 is obtained, followed by SFC separation.

[0123] [ka] The present invention further includes any variation of the process, for example using intermediate products available at any stage thereof as starting materials and carrying out the remaining steps, forming the starting materials in situ under the reaction conditions, or using the reaction components in the form of their salts or optically pure materials. The compounds and intermediates of the present invention can be converted into each other according to methods generally known to those skilled in the art.

[0124] Pharmacology and utility In one aspect, the present invention provides a compound of formula (I) or a subformula thereof, or a pharma- ceutically acceptable salt thereof, useful in therapy, in particular for treating or preventing a disease or condition mediated by NSD2.

[0125] In another aspect, the present invention provides the use of a compound of formula (I) or a subformula thereof, or a pharma- ceutically acceptable salt thereof, for treating a disease or condition that benefits from or is treatable by inhibition of NSD2, and for the manufacture of a medicament for treating a disease or condition that is treatable by inhibition of NSD2.

[0126] Examples of diseases or conditions that are mediated by NSD2 or that would benefit from or be treatable by inhibition of NSD2 include, but are not limited to, breast cancer, cervical cancer, skin cancer (particularly cutaneous squamous cell carcinoma), ovarian cancer, gastric cancer, prostate cancer, pancreatic cancer, lung cancer, hepatocellular carcinoma, head and neck cancer, peripheral nerve sheath tumor, osteosarcoma, multiple myeloma, neuroblastoma, leukemia (particularly acute lymphoblastic leukemia), non-Hodgkin's lymphoma (particularly mantle cell lymphoma), and pulmonary arterial hypertension.

[0127] Pharmaceutical Compositions, Dosages, and Administration In another aspect, the present invention provides a pharmaceutical composition comprising a compound of the present invention, or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable carrier.

[0128] In further embodiments, the composition comprises at least two pharma- ceutically acceptable carriers, such as those described herein. Pharmaceutical compositions can be formulated for specific routes of administration, such as oral administration, parenteral administration (e.g., injection, infusion, transdermal, or topical administration), and rectal administration. Topical administration can also involve inhalation or intranasal application. Pharmaceutical compositions of the present invention can be made in solid form (including, but not limited to, capsules, tablets, pills, granules, powders, or suppositories), or in liquid form (including, but not limited to, solutions, suspensions, or emulsions). Tablets can be either film-coated or enteric-coated according to methods known in the art. Typically, pharmaceutical compositions are tablets or gelatin capsules that contain the active ingredient along with one or more of the following: a) diluents, such as lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, and / or glycine; b) lubricants, such as silica, talcum, stearic acid, its magnesium or calcium salts, and / or polyethylene glycol, also for tablets. c) if desired, binders, such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, and / or polyvinylpyrrolidone; d) disintegrants, such as starch, agar, alginic acid or its sodium salt, or effervescent mixtures, and e) Absorbents, colouring agents, flavouring agents and sweetening agents.

[0129] In another embodiment, the compounds of the present invention may be combined with other therapeutic agents, such as other anti-cancer agents, anti-allergy agents, anti-nausea agents (or anti-emetics), analgesics, cytoprotective agents, and combinations thereof.

[0130] In one embodiment, the other therapeutic agent is an anti-cancer or chemotherapeutic agent. Examples of anti-cancer agents contemplated for use in the combination therapy of the present invention include erlotinib, bortezomib, fulvestrant, sunitib, imatinib mesylate, letrozole, finasunate, platins such as oxaliplatin, carboplatin, and cisplatin, finasunate, fluorouracil, rapamycin, leucovorin, lapatinib, lonafamib, sorafenib, gefitinib, capmutotecin, topotecan, bryostatin, adezelesin, anthracyclines, carzelesin, bizelin, dolastine, dasertin ... vincristine, auristatins, duocarmycins, eleutherobin, taxols such as paclitaxel or docetaxel, cyclophosphamide, doxorubicin, vincristine, prednisone or prednisolone, other alkylating agents such as mechlorethamine, chlorambucil, and ifosfamide, antimetabolites such as azathioprine or mercaptopurine, other microtubule inhibitors (vinca alkaloids such as vincristine, vinblastine, vinorelbine, vindesine, and taxanes), podophyllotoxins (etoposide, teniposide, etoposide phosphate, and epipodophyllotoxin), topoisomerase inhibitors, other cytotoxins, such as actinomycin, daunorubicin, barbicin, idarubicin, edrecolomab, epirubicin, bleomycin, plicamycin, mitomycin, and other anticancer antibodies (cetuximab, bevacizumab, ibritumomab, abagovomab, adecatumumab, afutuzumab, alacizumab, alemtuzumab, anatumomab, apolizumab, bavituximab, belimumab, bivatuzumab mertansine, blinatumomab, bleomycin ... ntuximab vedotin, cantuzumab mertansine, catumazomab, cetuximab, sitatuzumab bogatox, cixutumumab, clivatuzumab tetraxetan, conatumumab, dacetuzumab, daclizumab, detumomab, ecromeximab, edrecolomab, elotuzumab, epratuzumab, ertumaxomab, etaracizumab, farletuzumab, figitumumab, fresolimumab, galiximab, gemtuzumab vedotin, gemtuzumab, ibritumomab tiuxetan, inotuzumab ozogamicin, intetumumab, ipilimumab,Iratumumab, labetuzumab, lexatuzumab, lintuzumab, lucatuzumab, rumilisimab, mapatumumab, matumumab, milatuzumab, mitumomab, nacolomab, tafenatox, naptumomab estafenatox, necitumumab, nimotuzumab, ofatumumab, olaratumab, oportuzumab monat oregovomab, panitumumab, pemtumomab, pertuzumab, pintumomab, pritumumab, ramucirumab, rilotumumab, lobatumumab, rituximab, sibrotuzumab, tacatuzumab tetraxetan, taplitumomab paptox, tenatumomab, ticilimumab, tigatuzumab, tositumomab, or 131 I-Tositumomab, Trastuzumab, Tremelimumab, Tuocotuzumab-Celmoleukin, Veltuzumab, Vicilizumab, Volosixumab, Votumumab, Zalutumumab, Zanolimumab, IGN-101, MDX-010, ABX-EGR, EMD72000, ior-t1, MDX-220, MRA, H-11scFv, huJ591, TriGem, TriAb, R3, MT-201, G-250, ACA-125, Onyvax-105, CD:-960, Cea-Vac, BrevaRexAR54, IMC-1C11, G These include, but are not limited to, antibody drug conjugates comprising any of the above agents (particularly the auristatins MMAE and MMAF, maytansinoids such as DM-1, calicheamicin, or various cytotoxins).

[0131] In another embodiment, the compound of the invention is selected from the group consisting of anastrozole (ARIMIDEX®), bicalutamide (CASODEX®), bleomycin sulfate (BLENOXANE®), busulfan (MYLERAN®), busulfan injection (BUSULFEX®, capecitabine (XELODA®), N4-pentoxycarbonyl-5-deoxy-5-fluorocytidine, carboplatin (PARAPLATIN®), carmustine (BiCNU®), chlorambucil (LE UKERAN®), cisplatin (PLATINOL), cladribine (LEUSTATIN®), cyclophosphamide (CYTOXAN® or NEOSAR®), cytarabine, cytosine arabinoside (CYTOSAR-U®), cytarabine liposome injection (DEPOCYT®), dacarbazine (DTIC-Dome®), dactinomycin (actinomycin D, COSMEGAN®), daunorubicin hydrochloride (CERUBIDINE®), quercetin (QQE®), cyclophosphamide (CYTOXAN® or NEOSAR ... Daunorubicin liposome injection (DAUNOXOME®), dexamethasone, docetaxel (TAXOTERE®), doxorubicin hydrochloride (ADRIAMYCIN®), RUBEX®), etoposide (VEPESID®), fludarabine phosphate (FLUDARA®), 5-fluorouracil (ADRUCIL®, EFUDEX®), flutamide (EULEXIN®), tezacitibine, gemcitabine (difluorooxycytidine), hydroxyurea (HYDREA®), idarubicin (IDAMYCIN®), ifosfamide (IFEX®), irinotecan (CAMPTOSAR®), L-asparaginase (ELSPAR®), leucovorin calcium, melphalan (ALKERAN®), 6-mercaptopurine (PURINETHOL®), methotrexate (FOLEX®), mitoxantrone (NOVANTRONE®), gemtuzumab ozogamicin (MYLOTARG®),Combined with another therapeutic agent selected from paclitaxel (TAXOL®), nab-paclitaxel (ABRAXANE®), phoenix (Yttrium 90 / MX-DTPA), pentostatin, polipheprosan 20 with carmustine implant (GLIADEL®), tamoxifen citrate (NOLVADEX®), teniposide (VUMON®), 6-thioguanine, thiotepa, tirapazamine (TIRAZONE®), injectable topotecan hydrochloride (HYCAMPTIN®), vinblastine (VELBAN®), vincristine (ONCOVIN®), and vinorelbine (NAVELBINE®).

[0132] In another embodiment, the compound of the present invention is combined with another therapeutic agent capable of inhibiting BRAF, MEK, CDK4 / 6, SHP-2, HDAC, EGFR, MET, mTOR, PI3K, or AKT, or a combination thereof. In a particular embodiment, the compound of the present invention is combined with another therapeutic agent selected from vemurafinib, debrafinib, LGX818, trametinib, MEK162, LEE011, PD-0332991, panobinostat, belinostat, romidepsin, cetuximab, gefitinib, erlotinib, lapatinib, panitumumab, vandetanib, INC280, everolimus, simolimus, BMK120, BYL719, or CLR457, or a combination thereof.

[0133] In another embodiment, the therapeutic agent for use with the compound of the present invention is selected based on the disease or condition being treated.For example, in the treatment of melanoma, the other therapeutic agent can be selected from aldesleukin (e.g., PROLEUKIN®), dabrafenib (e.g., TAFINLAR®), dacarbazine, recombinant interferon alpha-2b (e.g., INTRON® A), ipilimumab, trametinib (e.g., MEKINIST®), pegylated interferon alpha-2b (e.g., PEGINTRON®, SYLATRON®), vemurafenib (e.g., ZELBORAF®), and ipilimumab (e.g., YERVOY®).

[0134] For the treatment of ovarian cancer, other therapeutic agents may be selected from doxorubicin hydrochloride (Adriamycin®), carboplatin (PARAPLATIN®), cyclophosphamide CYTOXAN®, NEOSAR®), cisplatin (PLATINOL®, PLATINOL-AQ®), doxorubicin hydrochloride liposomal (DOXIL®, DOX-SL®, EVACET®, LIPODOX®), gemcitabine hydrochloride (GEMZAR®), topotecan hydrochloride (HYCAMTIN®), and paclitaxel (TAXOL®).

[0135] For the treatment of thyroid cancer, the other therapeutic agent may be selected from doxorubicin hydrochloride (Adriamycin®), cabozantinib-S-malate (COMETRIQ®), and vandetanib (CAPRELSA®).

[0136] For the treatment of colon cancer, other therapeutic agents may be selected from fluorouracil (e.g., ADRUCIL®, EFUDEX®, FLUOROPLEX®), bevacizumab (AVASTIN®), irinotecan hydrochloride (CAMPTOSTAR®), capecitabine (XELODA®), cetuximab (ERBITUX®), oxaliplatin (ELOXATIN®), leucovorin calcium (WELLCOVORIN®), regorafenib (STIVARGA®), panitumumab (VECTIBIX®), and ZIV-aflibercept (ZALTRAP®).

[0137] For the treatment of lung cancer, other therapeutic agents include methotrexate, methotrexate LPF (e.g., FOLEX®, FOLEXPFS®, Abitrexate®, MEXATE®, MEXATE-AQ®), paclitaxel (TAXOL®), paclitaxel albumin-stabilized nanoparticle formulation (ABRAXANE®), afatinib dimaleate (GILOTRIF®), pembrolizumab (PEN), rifametinib (RIA), rifametinib riboflavin ... It may be selected from trexedine disodium (ALIMTA®), bevacizumab (AVASTIN®), carboplatin (PARAPLATIN®), cisplatin (PLATINOL, PLATINOL-AQ®), crizotinib (XALKORI®), erotinib hydrochloride (TARCEVA®), gefitinib (IRESSA®), and gemcitabine hydrochloride (GEMZAR®).

[0138] For the treatment of pancreatic cancer, other therapeutic agents may be selected from fluorouracil (ADRUCIL®), EFUDEX®, FLUOROPLEX®), erlotinib hydrochloride (TARCEVA®), gemcitabine hydrochloride (GEMZAR®), and mitomycin or mitomycin C (MITOZYTREXTM, MUTAMYCIN®).

[0139] For the treatment of cervical cancer, other therapeutic agents may be selected from bleomycin (BLENOXANE®), cisplatin (PLATINOL®, PLATINOL-AQ®), and topotecan hydrochloride (HYCAMTIN®).

[0140] For the treatment of head and neck cancer, the other therapeutic agent may be selected from methotrexate, methotrexate LPF (e.g., FOLEX®, FOLEX PFS®, Abitrexate®, MEXATE®, MEXATE-AQ®), fluorouracil (ADRUCIL®, EFUDEX®, FLUOROPLEX®), bleomycin (BLENOXANE®), cetuximab (ERBITUX®), cisplatin (PLATINOL®, PLATINOL-AQ®), and docetaxel (TAXOTERE®).

[0141] For the treatment of leukemia, including chronic myelomonocytic leukemia (CMML), other therapeutic agents may be selected from bosutinib (BOSULIF®), cyclophosphamide (CYTOXAN®, NEOSAR®), cytarabine (CYTOSAR-U®, TARABINE PFS®), dasatinib (SPRYCEL®), imatinib mesylate (GLEEVEC®), ponatinib (ICLUSIG®), nilotinib (TASIGNA®), and omacetaxine mepesuccinate (SYNRIBO®).

[0142] In another aspect, the present invention provides pharmaceutical compositions comprising at least one compound of the present invention (e.g., a compound of Formula (I) or subformulas thereof), or a pharma- ceutically acceptable salt thereof, alone or in combination with other anti-cancer agents, in association with a pharma- ceutically acceptable carrier suitable for administration to a human or animal subject.

[0143] In combination therapy, the compositions are formulated together as a combination therapy or as separate compositions. The compounds of the present invention and other therapeutic agents can be manufactured and / or formulated by the same or different manufacturers. The structures of therapeutic agents identified by code numbers, generic names, or trade names can be obtained from the actual version of the standard compendium "The Merck Index" or databases such as Patents International (e.g., IMS World Publications). Other therapeutic agents that can be used in combination with the compounds of the present invention can be prepared and administered as described in the art, for example in the documents listed above.

[0144] Optionally, the pharmaceutical composition may include a pharma- ceutically acceptable carrier as described above. The pharmaceutical composition or combination of the present invention may be a unit dose of about 0.5-1000 mg of active ingredient(s), or about 1-500 mg, or about 1-250 mg, or about 1-150 mg, or about 0.5-100 mg, or about 1-50 mg of active ingredient for a subject of about 50-70 kg. The therapeutically effective dose of the compound, pharmaceutical composition, or combination thereof depends on the species, weight, age, and individual condition of the subject to be treated, the disorder or disease being treated, or its severity. A physician, clinician, or veterinarian of ordinary skill can readily determine the effective amount of each of the active ingredients required to prevent, treat, or inhibit the progression of the disorder or disease. The above-mentioned dosage characteristics are demonstrable in vitro and in vivo tests, advantageously using mammals, e.g., mice, rats, dogs, monkeys, or isolated organs, tissues, and preparations thereof. The compounds of the invention can be applied in vitro in the form of solutions, for example aqueous solutions, and in vivo enterally, parenterally, advantageously intravenously, for example as a suspension or in aqueous solution.

[0145] In another aspect, the present invention provides a method for treating a human or animal subject suffering from a cell proliferative disease such as cancer, comprising administering to the subject a therapeutically effective amount of a compound of the present invention or a pharma- ceutically acceptable salt thereof alone or in combination with another anti-cancer agent. In a combination therapy, the compound of the present invention and another anti-cancer agent(s) can be administered simultaneously, concurrently, or sequentially without being limited to a specific time, and such administration provides a therapeutically effective level of the two compounds in the patient's body. Furthermore, the compound of the present invention and another therapeutic agent can be incorporated into the combination therapy (i) before the release of the combination product to the physician (e.g., in the case of a kit containing the compound of the present invention and another therapeutic agent), (ii) immediately before administration by the physician himself (or under the guidance of the physician), or (iii) in the patient himself, for example, during the sequential administration of the compound of the present invention and another therapeutic agent.

[0146] In one embodiment, the compound of the present invention and the other anticancer drug(s) are generally administered sequentially in any order by injection or oral administration. The administration regimen may vary depending on the stage of the disease, the physical condition of the patient, the safety profile of each drug, and the tolerability of each drug, as well as other criteria known to the attending physician and the physician(s) administering the combination. The compound of the present invention and the other anticancer drug(s) may be administered minutes, hours, days, or even weeks apart from each other, depending on the particular cycle used for treatment. Furthermore, the cycle may include administering one drug more frequently than the other during the treatment cycle, and at different doses for each administration of the drug.

[0147] In yet another embodiment, the compounds of the present invention may be combined with other anti-cancer agents, anti-allergy agents, anti-nausea agents (or anti-emetics), analgesics, cytoprotective agents, and combinations thereof.

[0148] In some cases, patients may experience allergic reactions to the compounds of the present invention and / or other anti-cancer drug(s) during or after administration. Therefore, to minimize the risk of allergic reactions, anti-allergy agents may be administered. Suitable anti-allergy agents include corticosteroids such as dexamethasone (e.g., DECADRON®), beclomethasone (e.g., BECLOVENT®), hydrocortisone (also known as cortisone, hydrocortisone sodium phosphate, hydrocortisone sodium phosphate, e.g., ALA-CORT®, hydrocortisone phosphate, Solu-CORTEF®, HYDROCORT Acetate®, and LANACORT®), prednisolone (e.g., DELTA-Cortel®, ORAPRED®, PEDIAPRED®, and PRELONE®), prednisone (e.g., DELTASONE, LIQUID RED®, DIACETATE®, and DIACETATE®), and DIACETATE®. RED®, METICORTEN®, and ORASONE®, methylprednisolone (6-methylprednisolone, methylprednisolone acetate, methylprednisone sodium succinate, e.g., DURALONE®, MEDRALONE®, MEDROL®, M-PREDNISOL®, and SOLU-MEDROL®), antihistamines (e.g., BENADRYL®, hydroxyzine, and cyproheptadine), bronchodilators, e.g., beta-adrenergic receptor agonists, albuterol (e.g., PROVENTIL®, terbutaline (BRETHINE®).

[0149] In other cases, patients may experience nausea during and after administration of the compounds of the present invention and / or other anticancer drug(s). Therefore, antiemetics may be administered to prevent nausea (upper stomach) and vomiting. Suitable antiemetics include aprepitant (EMEND®), ondansetron (ZOFRAN®), granisetron HCl (KYTRIL®), lorazepam (ATIVAN®), dexamethasone (DECADRON®), prochlorperazine (COMPAZINE®), casopitant (REZONIC® and Zunrisa®), and combinations thereof.

[0150] In still other cases, medications to alleviate pain experienced during treatment are prescribed to make patients more comfortable. Common painkillers available over the counter, such as TYLENOL®, are often used. Opioid analgesics, such as hydrocodone / paracetamol or hydrocodone / acetaminophen (e.g., VICODIN®), morphine (e.g., ASTRAMORPH® or AVINZA®), oxycodone (e.g., OXYCONTIN® or PERCOCET®), oxymorphone hydrochloride (OPANA®), and fentanyl (e.g., DURAGESIC®), are also useful for moderate or severe pain.

[0151] Additionally, cytoprotectants (neuroprotectants, free radical scavengers, cardioprotectants, anthracycline extravasation neutralizers, nutrients, etc.) may be used as adjunctive therapy to protect normal cells from treatment toxicity and limit organ toxicity. Suitable cytoprotectants include amifostine (ETHYOL®), glutamine, dimesna (TAVOCEPT®), mesna (MESNEX®), dexrazoxane (ZINECARD® or TOTECT®), xaliproden (XAPRILA®), and leucovorin (also known as calcium leucovorin, citrovorum factor, and folic acid).

[0152] In yet another embodiment, the compounds of the present invention can be used in conjunction with known therapeutic processes, such as administration of hormones or in radiation therapy. In certain instances, the compounds of the present invention can be used as radiosensitizers, particularly for the treatment of tumors that exhibit poor sensitivity to radiation therapy.

[0153] In yet another aspect, the present invention provides a kit comprising one or more compounds of the present invention and another therapeutic agent as described above. A representative kit comprises (a) a compound of formula (I) or a subformula thereof, or a pharma- ceutically acceptable salt thereof, and (b) at least one other therapeutic agent, such as those described above, and such a kit may further comprise a package insert or other labeling, including administration instructions. The kit of the present invention may be used, for example, to administer different dosage forms for oral and parenteral dosage forms, to administer two or more separate pharmaceutical compositions at different dosage intervals, or to titrate separate compositions together, and at least one pharmaceutical composition comprises a compound of formula (I) or a subformula thereof.

[0154] Temperatures are given in degrees Celsius. The structure of final products, intermediates and starting materials is confirmed by standard analytical methods, e.g., microanalysis and spectroscopic characteristics, e.g., MS, IR, NMR. The abbreviations used are conventional in the art.

[0155] All starting materials, building blocks, reagents, acids, bases, dehydrating agents, solvents, and catalysts utilized to synthesize the compounds of the present invention are either commercially available or can be prepared by any of the organic synthesis methods known to those skilled in the art ((Houben-Weyl 4th Ed. 1952, Methods of Organic Synthesis, Thieme, Volume 21). Unless otherwise indicated, starting materials are generally available from commercial sources.

[0156] The examples herein are merely illustrative of the present invention and do not limit the scope of the invention as claimed. In addition, the compounds of the present invention can be prepared by organic synthesis methods known to those skilled in the art, as shown in the following examples. If desired, conventional protecting groups are used to protect reactive functional groups according to standard practices, see, for example, TW Greene and PG M Huts in "Protecting Groups in Organic Synthesis", John Wiley and Sons, 1991.

[0157] Abbreviation Abbreviations used herein are defined as follows: "1x" for 1st, "2x" for 2nd, "3x" for 3rd, "°C" for degrees Celsius, "aq" for aqueous, "FCC" for flash column chromatography, "eq" for equivalent or equivalents, "g" for gram or grams, "mg" for milligram or milligrams, "L" for liter or liters, "mL" for milliliter or milliliters, "μL" for microliter or microliters, "N" for normal, "M" for mole, "nM" for nanomole, "mol" for mole or moles, "mmol" for millimole or millimoles, "min" for minute or minutes, "h" or "hrs" for hour or hours, "RT" for room temperature, "ON" for overnight, "atm" for atmosphere, "psi" for pounds per square inch, "conc." for concentrate, "sat" or "sat'd" for saturated, "MW" for molecular weight, "mw" or "μwave" for microwave, "mp" for melting point, "Wt" for weight, "MS" or "Mass" for mass spectrometry Spec', 'ESI' for electrospray ionization mass spectrometry, 'HR' for high resolution, 'HRMS' for high resolution mass spectrometry, 'LCMS' or 'LC-MS' for liquid chromatography mass spectrometry, 'HPLC' for high pressure liquid chromatography, 'RP HPLC' for reversed phase HPLC, 'TLC' or 'tlc' for thin layer chromatography, 'NMR' for nuclear magnetic resonance spectroscopy, 'nOe' for nuclear Overhauser effect spectroscopy, and ' 1"H", "δ" for delta, "s" for singlet, "d" for doublet, "t" for triplet, "q" for quartet, "m" for multiplet, "br" for broad, "Hz" for Hertz, "ee" for enantiomeric excess, as well as "α", "β", "R", "r", "S", "s", "E", and "Z" are stereochemical designations well known to those of skill in the art.

[0158] The abbreviations used herein below have the corresponding meanings. [Table 1-1] [Table 1-2]

[0159] High performance liquid chromatography (HPLC) was performed using an Agilent 1260 HPLC system (Santa Clara, CA). The analytical column was a reversed-phase Phenomenex Kinetex C18-2.6 μm, 4.6×50 mm. Gradient elution (flow rate 2.0 mL / min) was used, starting with 5% methanol / 95% water and progressing to 95% methanol / 5% water over 10 min. All solvents contained 0.1% formic acid (FA). Compounds were detected by ultraviolet (UV) absorption at 214, 254, and 300 nm. HPLC solvents were purchased from Sigma Aldrich (St. Louis, MO).

[0160] Mass spectrometry was performed on an Agilent system (Agilent 1260 HPLC and Agilent 6130 mass spectrometer detector; column: Phenomenex Kinetex 2.6um C18, column size 4.6 x 50 mm, column temperature 40 °C, gradient: 5-95% methanol in water with 0.1% FA over 2 min; flow rate 2.0 mL / min (or 5-50% polar gradient over 2.0 min, or 50-95% non-polar gradient over 2.0 min); mass spectrometer molecular weight scan range 100-1000 or 100-1500; capillary voltage 4000 V). All masses are reported as the mass of the protonated parent ion unless otherwise indicated.

[0161] Nuclear magnetic resonance (NMR) analyses were performed using a Bruker 400 MHz NMR. Spectral references were either TMS or known solvent chemical shifts.

[0162] Chiral Preparative HPLC Method Used for Purification of Examples SFC chiral screening was performed on a Thar Instruments Investigator system. The Thar Investigator system consists of: ●ALIAS automatic sampler ●Thar Liquid Delivery Module (0~10mL / min) ●Thar SFC 10 position column oven ●Waters 2998 PDA ●Thar automatic back pressure regulator

[0163] All Thar components are part of the SuperPure Discovery series. The system was run at 3.0 mL / min and maintained at 38° C. The system backpressure was set at 100 bar. Each sample was screened through a battery of ten 5 μM columns: ●5μm 4.6×150mm ChiralPak AD ●5μm 4.6×150mm ChiralCel OD ●5μm 4.6×150mm ChiralCel OJ ●5μm 4.6×150mm ChiralPak AS ●5μm 4.6×250mm ChiralPak AY ●5μm 4.6×250mm ChiralCel OZ ●5μm 4.6×150mm ChiralPak IC ●5μm 4.6×150mm ChiralPak IG ●5μm 4.6×250mm Regis Whelk-O1 ●5μm 4.6×250mm ChromegaChiral CC4

[0164] The system ran a gradient from 5% co-solvent to 50% co-solvent in 9 min, followed by a 10 min hold at 50% co-solvent, a switch back to 5% co-solvent and a 0.5 min hold at the initial conditions. Between each gradient there was a 4 min equilibration procedure where 5% co-solvent was run on the next column being screened. Typical solvents screened were MeOH, EtOH, IPA, MeOH + 0.5% NH 3 , EtOH+0.5% NH 3 , IPA + 0.1% NH 3 Once separation is found using one of the gradient methods, an isocratic method can be developed and, if necessary, scaled up for separation on the Thar Prep80 system.

[0165] Intermediates Intermediate A: Methyl 2-(3,4-difluorophenyl)-5-fluoroisonicotinate [ka] 1,4-Dioxane (Pd(dppf)Cl 2 Methyl 2-bromo-5-fluoroisonicotinate (110 g, 470.04 mmol, 1.0 equiv.), (3,4-difluorophenyl)boronic acid (111.34 g, 705.06 mmol, 1.5 equiv.), Pd(dppf)Cl in 1 L 2(8.60 g, 11.75 mmol, 0.025 equiv.) and potassium carbonate (129.92 g, 940.08 mmol, 2.0 equiv.) were added to a mixture of 2 The mixture was heated to 100° C. under reduced pressure for 16 h. 2 The mixture was diluted with 200 mL of HO and 1.3 L of EtOAc and stirred for 15 min. The organic layer was separated, washed with brine (500 mL) and added Na 2 SO 4 The mixture was dried at 40° C., concentrated and the residue was purified by combi-flash (5% to 50% EtOAc in PE) and trituration (PE:EA=50 / 1, 100 mL) to give methyl 2-(3,4-difluorophenyl)-5-fluoroisonicotinate (Intermediate A). 1 H NMR (CDCl 3 400MHz):δ 8.56(d,J=2.0Hz,1H),8.05(d,J=5.6Hz,1H),7.79(ddd,J=2.4,8.4,10.8Hz,1H),7.64(td,J=2.8,8.8Hz,1H),7.26-7.11(m,1H),3.94(s,3H).

[0166] Intermediate B [ka] To a solution of the compound 9H-purin-6-amine (15 g, 111 mmol) in THF (500 mL) was added di-tert-butyl dicarbonate (Boc 2 HO) (96.91 g, 444 mmol) and DMAP (1.36 g, 11.1 mmol) were added. The resulting mixture was stirred at room temperature for 16 h. THF was removed in vacuo. The residue was dissolved in EtOAc (400 mL) and HCl solution (H 2 The organic layer was partitioned between Na 2 SO 4 and concentrated to give tert-butyl 6-(bis(tert-butoxycarbonyl)amino)-9H-purine-9-carboxylate (Intermediate B-1).

[0167] To a solution of tert-butyl 6-(bis(tert-butoxycarbonyl)amino)-9H-purine-9-carboxylate (Intermediate B-1) (48 g, 110 mmol) in MeOH (500 mL), saturated NaHCO 3 A solution (250 mL) was added. The reaction mixture was stirred at 60° C. for 2 h. MeOH was removed in vacuo. The residue was extracted with a solution of MeOH:DCM (1:10) (300 mL×7). The combined organic layers were washed with H 2 Wash with O (150 mL) and add Na 2 SO 4 The mixture was dried at 40° C. and concentrated. The residue was washed with a solution of EtOAc:PE (1:10) (100 mL) and filtered. The filter cake was dried in vacuum to give intermediate B. 1 H NMR (400MHz, DMSO-d 6 )δ8.70(s,1H),8.48(s,1H),1.36(s,18H). LC-MS:[M+H)] + =336.2.

[0168] Intermediate C: Methyl 5-fluoro-2-(3-fluorophenyl)isonicotinate [ka] Methyl 5-fluoro-2-(3-fluorophenyl)isonicotinate (Intermediate C) was prepared using a procedure similar to that of Intermediate A. 1 H-NMR (400MHz, CDCl 3 )δ 8.66(s,1H),8.18(d,J=6.4Hz,1H),7.76-7.72(m,2H),7.48-7.42(m,1H),7.15-7.11(m,1H),4.01(s,3H).

[0169] Intermediate D: Methyl 5-fluoro-2-(3-fluoro-4-methoxyphenyl)isonicotinate [ka] Methyl 2-bromo-5-fluoroisonicotinate (1.0 g, 4.27 mmol), (3-fluoro-4-methoxyphenyl)boronic acid (1.089 g, 6.41 mmol), Cs in dioxane (10 mL). 2 CO 3 (2.78 g, 8.55 mmol), and PdCl 2 (dppf)·CH 2 Cl 2 A suspension of the adduct (0.349 g, 0.427 mmol) was stirred under Ar at 100° C. for 2 h. The mixture was then diluted with water and extracted with EtOAc, and the organic layer was washed with Na 2 SO 4 The residue was purified by CombinFlash (PE / EA, EA: 20-40%, 50 min) to give methyl 5-fluoro-2-(3-fluoro-4-methoxyphenyl)isonicotinate (Intermediate D). 1 H NMR (400MHz, DMSO-d 6 )δ8.79(t,1H),8.24(t,1H),7.95-7.87(m,2H),7.27(t,3H),3.92(d,6H). LC-MS:[M+H] + =279.9.

[0170] Intermediate F-2: 2-(2-(difluoromethyl)-4-fluorophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane [ka] To a solution of 2-bromo-5-fluorobenzaldehyde (8 g, 39.4 mmol) in DCM (30 mL) was added DAST (10.41 mL, 79 mmol) at 0° C., and the mixture was then cooled to 5° C. with N 2 The mixture was stirred at room temperature under reduced pressure for 18 hours. The reaction mixture was washed with water, and the organic layer was extracted with Na 2 SO 4 The mixture was dried over 1000 ml of ethyl acetate, filtered and concentrated in vacuo. The residue was purified by CombinFlash (elution gradient: 0% to 10% EtOAc in n-hexane, 30 min) to give 1-bromo-2-(difluoromethyl)-4-fluorobenzene (Intermediate F-1). LC-MS: [M+H]+ =225.0,227.0.

[0171] 1-Bromo-2-(difluoromethyl)-4-fluorobenzene (Intermediate F-1) (2.2 g, 9.78 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane (2.98 g, 11.73 mmol), and KOAc (1.919 g, 19.56 mmol) in dioxane (30 mL), and PdCl 2 (dppf)·CH 2 Cl 2 A suspension of the adduct (0.798 g, 0.978 mmol) was added to N 2 The mixture was stirred under atmospheric pressure at 110° C. for 3 hours. The mixture was diluted with water, extracted with EtOAc, and the organic layer was washed with Na 2 SO 4 The mixture was dried at 40° C., filtered and concentrated in vacuo. The residue was purified by CombinFlash (elution gradient: 0% to 10% EtOAc in PE, 30 min) to give 2-(2-(difluoromethyl)-4-fluorophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (Intermediate F-2) as a colorless syrup. LC-MS: [M+H] + =272.3.

[0172] Intermediate L-2: 2-(2-(difluoromethyl)-4,5-difluorophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane [ka] 1-Bromo-2-(difluoromethyl)-4,5-difluorobenzene (Intermediate L-1) and 2-(2-(difluoromethyl)-4,4,5,5-difluorophenyl)-4,5-tetramethyl-1,3,2-dioxaborolane (Intermediate L-2) were prepared following procedures similar to the preparation of Intermediates F-1 and F-2, respectively.

[0173] Intermediate L-1: 1 H NMR (CDCl 3,400MHz):δ 7.54-7.45(m,2H),6.97-6.70(t,J1=54.8Hz,J2=109.6Hz,1H).

[0174] Intermediate L-2: 1 H NMR (CDCl 3 ,400MHz):δ 7.75-7.66(m,1H),7.57-7.50(m,1H),7.47-7.14(m,1H),1.36(s,12H).

[0175] Intermediate O-1: 6-bromo-2-chloro-3-fluoropyridine [ka] 6-Bromo-2-chloro-3-fluoropyridine (3.0 g, 0.014 mol, 1.0 equiv.) and i-Pr in anhydrous THF (30 mL). 3 O 3 To a solution of B (4.0 g, 0.021 mol, 1.5 equiv) was added n-BuLi (9 mL, 0.021 mol, 2.5 M sol, 1.5 equiv) dropwise at -78 °C. The resulting mixture was stirred at -78 °C for 1 h and then warmed to 20 °C for an additional 1 h. The reaction was quenched with saturated NH 4 The mixture was quenched with Cl (50 mL), acidified to pH = 4 with concentrated HCl, and stirred at 20 °C for 1 h. The mixture was extracted with ethyl acetate (50 mL x 3). The combined organic layers were washed with Na 2 SO 4 and concentrated in vacuo to give 6-bromo-2-chloro-3-fluoropyridine (Intermediate O-1). LC-MS: [M+H] + =176.0.

[0176] Intermediate EO was prepared following a procedure similar to the preparation of intermediate A from the reaction of methyl 2-bromo-5-fluoroisonicotinate with the corresponding boronic acid or ester. [Table 2-1] [Table 2-2]

[0177] Intermediate 1-1: 5-fluoro-2-(3-fluoro-4-methoxyphenyl)isonicotinaldehyde [ka] 2-Bromo-5-fluoroisonicotinaldehyde (8 g, 27.5 mmol), (3-fluoro-4-methoxyphenyl)boronic acid (4.67 g, 27.5 mmol), PdCl in dioxane (50 mL) 2 (dppf)·CH 2 Cl 2 A suspension of the adduct (1.121 g, 1.373 mmol) and cesium carbonate (13.42 g, 41.2 mmol) was stirred at 100° C. under nitrogen atmosphere for 3 h. The reaction mixture was filtered and the filtrate was concentrated in vacuum to give a black residue. The residue was redissolved in DCM (60 mL), washed with water (20 mL), brine (20 mL), dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuum. The residue was purified by silica gel column chromatography (EtOAc in hexane and DCM, EtOAc / DCM / hexane=8 / 20 / 100) to give 5-fluoro-2-(3-fluoro-4-methoxyphenyl)isonicotinaldehyde (Intermediate 1-1). 1 H NMR (400 MHz, CDCl 3 )δ 10.51(d,J=2.1Hz,1H),8.77(d,J=1.9Hz,1H),8.07(dd,J=5.3,2.1Hz,1H),7.85(dd,J=12 .5,2.4Hz,1H),7.78(d,J=8.6Hz,1H),7.11(td,J=8.6,2.2Hz,1H),4.01(d,J=2.2Hz,3H). LC-MS:[M+H] + =249.9.

[0178] Intermediate 22-7: tert-butyl (3-(cyclopropoxymethyl)piperidin-3-yl)carbamate [ka] To a solution of 1-(tert-butyl) 3-ethylpiperidine-1,3-dicarboxylate (70.0 g, 272 mmol, 1.0 equiv.) in THF (1.4 L), LiHMDS (435 mL, 435 mmol, 1.6 equiv., 1 M in THF) was added N 2 The mixture was added dropwise at −70° C. under atmospheric pressure. After stirring at −78° C. for 1 h, ((chloromethoxy)methyl)benzene (56.0 g, 354 mmol, 1.3 equiv.) was added at −78° C. The resulting mixture was warmed to 15-20° C. and stirred for 16 h. The reaction was quenched with water (800 mL) and extracted with EtOAc (800 mL×3). The combined organic layers were washed with anhydrous Na 2 SO 4 The residue was purified by Combi Flash (10% EtOAc in PE) to give 1-(tert-butyl) 3-ethyl 3-((benzyloxy)methyl)piperidine-1,3-dicarboxylate (22-1). 1 H NMR (400 MHz, CDCl 3 )δ ppm 7.20-7.40(5H,m),4.50(2H,q,J=12.8Hz),4.10-4.20(2H,m),3.65-3.85(2H,m),3.45-3.60(2H,m),3.3 0-3.45(2H,m),1.90-2.10(1H,m),1.65-1.75(1H,m),1.50-1.60(2H,m),1.45(9H,s),1.20-1.30(3H,m). LC-MS:[M+H-100] + =278.1.

[0179] A mixture of 1-(tert-butyl) 3-ethyl 3-((benzyloxy)methyl)piperidine-1,3-dicarboxylate (22-1) (50.0 g, 132 mmol, 1.0 equiv.) and Pd / C (5.00 g, 10% wet) in dioxane / 2N HCl (1 L, v / v=1 / 1) was diluted with H 2 The mixture was stirred under a balloon at 20° C. for 16 h. The mixture was then filtered and the filtrate was concentrated in vacuo. The pH of the residue was adjusted with NaHCO 3The mixture was adjusted to 8-9 with aqueous solution of 1,2-dichlorophenyl ether. Then, THF (500 mL) and CbzCl (45.0 g, 265 mmol, 2.0 equiv.) were added. The resulting mixture was stirred at 5-10 °C for 24 h. The mixture was extracted with EtOAc (500 mL × 3). The combined organic layer was washed with anhydrous Na 2 SO 4 The residue was purified by Combi Flash (30% EtOAc in PE) to give 1-benzyl 3-ethyl 3-(hydroxymethyl)piperidine-1,3-dicarboxylate (22-2). 1 H NMR (400 MHz, CDCl 3 )δ ppm 7.30-7.40(5H,m),5.05-5.20(2H,s),4.05-4.20(2H,m),2.60-4.05(7H,m),1.55-1.95(4H,m),1.15-1.30(3H,m).

[0180] To a solution of 1-benzyl 3-ethyl 3-(hydroxymethyl)piperidine-1,3-dicarboxylate (22-2) (31.0 g, 96.5 mmol, 1.0 equiv.) and Py (15.3 g, 193 mmol, 2.0 equiv.) in DCM (300 mL) was added Tf 2 O (57.0 g, 193 mmol, 2.0 equiv.) 2 The mixture was then warmed to 10-15°C and stirred for 1 h. The reaction was quenched with water (500 mL) and extracted with DCM (500 mL x 2). The combined organic layer was washed with water (500 mL x 3) and diluted with anhydrous Na 2 SO 4 The residue was purified by Combi Flash (10% EtOAc in PE) to give 1-benzyl 3-ethyl 3-((((trifluoromethyl)sulfonyl)oxy)methyl)piperidine-1,3-dicarboxylate (22-3). 1 H NMR (400 MHz, CDCl 3)δ ppm 7.30-7.45(5H,m),5.05-5.25(2H,s),4.45-4.75(2H,m),4.05-4.25(2H,m),3.60-3.85(2H,m), 3.20-3.40(3H,m),1.95-2.10(1H,m),1.75-1.90(1H,m),1.60-1.75(2H,m),1.15-1.30(3H,m).

[0181] To a solution of compound cyclopropanol (3.20 g, 55.1 mmol, 1.0 equiv) in THF (100 mL) was added NaH (60% in mineral oil, 2.70 g, 66.1 mmol, 1.2 equiv) at 0 °C. After stirring at 10-15 °C for 30 min, a solution of 1-benzyl 3-ethyl 3-((((trifluoromethyl)sulfonyl)oxy)methyl)piperidine-1,3-dicarboxylate (22-3) (27.0 g, 60.6 mmol, 1.1 equiv) in THF (100 mL) was added at 0 °C. The resulting mixture was stirred at 10-15 °C for 3 h. The reaction was quenched with water (500 mL) and extracted with EtOAc (500 mL × 2). The combined organic layer was washed with water (500 mL × 3) and diluted with anhydrous Na 2 SO 4 The mixture was dried at 40° C. and concentrated in vacuo. The residue was purified by Combi Flash (20% EtOAc in PE) to give 1-benzyl 3-ethyl 3-(cyclopropoxymethyl)piperidine-1,3-dicarboxylate (22-4). 1 H NMR (400 MHz, CDCl 3 )δ ppm 7.25-7.45(5H,m),5.05-5.25(2H,s),4.05-4.20(2H,m),3.86(1H,d,J=13.2Hz),3.30-3.60(5H,m) ,3.10-3.25(1H,m),1.90-2.05(1H,m),1.50-1.70(3H,m),1.95(3H,t,J=6.8Hz),0.30-0.55(4H,m).

[0182] MeOH / THF / H 21-Benzyl 3-ethyl 3-(cyclopropoxymethyl)piperidine-1,3-dicarboxylate (22-4) (10.0 g, 27.6 mmol, 1.0 equiv.) and LiOH.H 2 A mixture of 2,4-dimethylformamide (11.6 g, 276 mmol, 10.0 equiv.) and 1,2-dimethylformamide (1.0 g, 276 mmol, 10.0 equiv.) was stirred at 20° C. for 16 h. The mixture was then concentrated in vacuo to remove THF and MeOH. The residue was diluted with water (300 mL) and extracted with EtOAc (200 mL). The pH of the aqueous layer was adjusted to 5 with 1N HCl, and then the aqueous layer was extracted with EtOAc (300 mL×3). The combined organic layers were washed with anhydrous Na 2 SO 4 Drying at rt and concentration in vacuo afforded 1-((benzyloxy)carbonyl)-3-(cyclopropoxymethyl)piperidine-3-carboxylic acid (22-5). 1 H NMR (400 MHz, CDCl 3 )δ ppm 7.25-7.45(5H,m),5.14(2H,s),3.79(1H,d,J=13.6Hz),3.15-3.65(6H,m),1.90-2.00(1H,m),1.50-1.80(3H,m),0.30-0.60(4H,m).

[0183] 1-(((benzyloxy)carbonyl)-3-(cyclopropoxymethyl)piperidine-3-carboxylic acid (22-5) (6.00 g, 18 mmol, 1.0 equiv.), DPPA (5.50 g, 19.8 mmol, 1.1 equiv.), and Et 3 A solution of N (3.60 g, 36 mmol, 2.0 equiv.) was dissolved in N 2 The reaction mixture was stirred at 80° C. for 2 h under reduced pressure. The reaction was then concentrated in vacuo. The residue was dissolved in THF (100 mL). KOH (3.03 g, 54 mmol, 3.0 equiv.) and water (10 mL) were then added. The reaction mixture was stirred at 15-20° C. for 4 h. Boc 2 O (5.90 g, 27 mmol, 1.5 equiv.) and K 2 CO 3(10.0 g, 72 mmol, 4.0 equiv.) was added. The reaction mixture was stirred at 15-20 °C for 16 h. The mixture was extracted with EtOAc (200 mL × 3). The combined organic layer was washed with anhydrous Na 2 SO 4 The residue was purified by Combi Flash (15% EtOAc in PE) to give benzyl 3-((tert-butoxycarbonyl)amino)-3-(cyclopropoxymethyl)piperidine-1-carboxylate (22-6). 1 H NMR (400 MHz, CDCl 3 )δ ppm 7.25-7.45(5H,m),5.05-5.25(2H,m),4.40-4.80(1H,brs),3.50-4.05(4H,m),3.00-3. 35(3H,m),2.15-2.55(1H,m),1.60-1.75(1H,m),1.35-1.55(11H,m),0.30-0.60(4H,m).

[0184] A mixture of benzyl 3-((tert-butoxycarbonyl)amino)-3-(cyclopropoxymethyl)piperidine-1-carboxylate (22-6) (4.90 g, 12.1 mmol) and Pd / C (500 mg, 10% wet) in MeOH (100 mL) was diluted with H 2 The mixture was stirred under a balloon for 1 h at 15-20° C. Then, the Pd / C was filtered off, and the filtrate was concentrated in vacuo to give tert-butyl (3-(cyclopropoxymethyl)piperidin-3-yl)carbamate (Intermediate 22-7). 1 H NMR (400 MHz, CDCl 3 )δ ppm 4.82(1H,brs),3.80(1H,d,J=10.0Hz),3.53(1H,d,J=9.6Hz),3.20-3.30(1H,m),3.01(1H,d,J=12.4H z),2.85-2.95(1H,m),2.55-2.65(2H,m),2.05-2.20(1H,brs),1.30-1.70(13H,m),0.35-0.60(4H,m).

[0185] Intermediate 47-6: tert-butyl (3-(2,2-difluoroethyl)piperidin-3-yl)carbamate [ka] To a solution of 1-benzyl 3-methylpiperidine-1,3-dicarboxylate (5 g, 18.03 mmol) in THF (70 mL) was added 1M LHMDS in THF (19.83 mL, 19.83 mmol) dropwise at -78°C for 30 min. The mixture was stirred at -78°C for another 30 min. 1,1-difluoro-2-iodoethane (6.92 g, 36.1 mmol) was then added. The reaction mixture was stirred at -78°C for 2 h. The temperature was then allowed to slowly rise to room temperature and the mixture was stirred at room temperature overnight. The reaction was quenched by adding water (10 mL). Most of the THF was removed under reduced pressure. The residue was extracted with ethyl acetate (30 mL x 2) and the combined organic phase was washed with water (20 mL), brine (20 mL), dried over anhydrous magnesium sulfate, filtered and concentrated. The crude product was purified by flash chromatography (elution gradient: 5% to 20% EtOAc in hexanes, 40 min) to give 1-benzyl 3-methyl 3-(2,2-difluoroethyl)piperidine-1,3-dicarboxylate (47-1). LC-MS: [M+H] + =341.9.

[0186] To a solution of 1-benzyl 3-methyl 3-(2,2-difluoroethyl)piperidine-1,3-dicarboxylate (47-1) (6 g, 17.58 mmol) in methanol (50 mL) was added lithium hydroxide (4.21 g, 176 mmol) in water (30 mL). The mixture was stirred at room temperature overnight and the solvent was removed. The residue was diluted with water (20 mL), then the mixture was washed with ethyl acetate (5 mL) and the aqueous layer was acidified to PH=3 with HCl. The mixture was then extracted with DCM (20 mL×3). The combined organic layers were dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo to give crude 1-((benzyloxy)carbonyl)-3-(2,2-difluoroethyl)piperidine-3-carboxylic acid (47-2). LC-MS: [M+H] + =328.1.

[0187] To a solution of 1-((benzyloxy)carbonyl)-3-(2,2-difluoroethyl)piperidine-3-carboxylic acid (47-2) (5 g, 15.28 mmol) in toluene (70 mL) was added TEA (4.26 mL, 30.6 mmol) and DPPA (3.70 mL, 16.80 mmol). The mixture was stirred at 100° C. for 2 h under nitrogen protection. The mixture was diluted with ethyl acetate (60 mL) and washed with water (20 mL), brine (60 mL). The organic phase was dried over anhydrous magnesium sulfate, filtered and concentrated to give crude benzyl 3-(2,2-difluoroethyl)-3-isocyanatopiperidine-1-carboxylate (47-3). LC-MS: [M+H] + =325.1.

[0188] To a solution of benzyl 3-(2,2-difluoroethyl)-3-isocyanatopiperidine-1-carboxylate (47-3) (5.7 g, 12.30 mmol) in 1,4-dioxane (25 mL) was added 6M HCl (25 mL, 150 mmol). The reaction mixture was stirred at 45° C. for 30 h. The mixture was cooled to room temperature and diluted with 20 mL of water. It was then washed with ethyl acetate (5 mL). The aqueous phase was basified with sodium hydroxide to adjust the pH value to 9. The mixture was then extracted with DCM (30 mL×3). The organic phases were combined, dried over anhydrous magnesium sulfate, filtered and concentrated to give crude benzyl 3-amino-3-(2,2-difluoroethyl)piperidine-1-carboxylate (47-4). LC-MS: [M+H]+=299.2.

[0189] To a solution of benzyl 3-amino-3-(2,2-difluoroethyl)piperidine-1-carboxylate (Intermediate 47-4) (4 g, 13.41 mmol) in DCM (50 mL) was added Boc anhydride (4.67 mL, 20.11 mmol) and DIPEA (7.03 mL, 40.2 mmol) with N 2The mixture was added at room temperature under atmospheric pressure. The mixture was stirred at 40° C. for 20 h. The mixture was cooled to room temperature, diluted with DCM (30 mL), washed with water (20 mL), brine (20 mL), dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo to give the crude product. The crude product was purified by flash chromatography (elution gradient: 5% to 20% EtOAc in hexanes, 30 min) to give benzyl 3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoroethyl)piperidine-1-carboxylate (47-5). LC-MS: [M+H] + =399.0.

[0190] A solution of benzyl 3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoroethyl)piperidine-1-carboxylate (47-5) (4 g, 10.04 mmol) in methanol (50 mL) was diluted with 10% Pd(OH) on carbon. 2 Wet (1.410 g, 0.502 mmol) was added and the mixture was stirred under a hydrogen balloon at room temperature for 3 hours. The reaction mixture was filtered and concentrated to give crude tert-butyl (3-(2,2-difluoroethyl)piperidin-3-yl)carbamate (Intermediate 47-6). LC-MS: [M+H] + =265.0.

[0191] Intermediate 78-5: Methyl tert-butyl 3-(methylcarbamoyl)piperidin-3-ylcarbamate [ka] H 2 (NH) in O (8 mL) 4 ) 2 CO 3 To a solution of (2.13 g, 22 mmol) was added another solution of 1-benzylpiperidin-3-one (500 mg, 2 mmol) in EtOH (8 mL). Then, trimethylsilyl cyanide (TMSCN) (0.74 mL, 5.5 mmol) was added dropwise at room temperature. The reaction mixture was stirred at 70° C. for 16 h. EtOH was removed in vacuo. The resulting mixture was extracted with EtOAc (20 mL×3). The organic layer was washed with H2 Wash with 20 mL of O and 20 mL of saline. 2 SO 4 The residue was recrystallized from EtOAc:PE (1:5) to give 7-benzyl-1,3,7-triazaspiro[4.5]decane-2,4-dione (78-1). 1 H NMR (400MHz, CD 3 OD)δ 7.46-7.17(m,5H),3.54(dd,J=29.8,13.2Hz,2H),2.90(d,J=10.8Hz,1H),2.69(d, J=11.2Hz,1H),2.32(d,J=11.2Hz,1H),2.08(t,J=10.6Hz,1H),1.88-1.59(m,4H). LC-MS(UV 214):(M+H) + =260.1

[0192] A mixture of 7-benzyl-1,3,7-triazaspiro[4.5]decane-2,4-dione (78-1) (1 g, 3.86 mmol) in 10 mL of 2.0 M aqueous KOH was heated at 120 °C for 16 h. The mixture was cooled to room temperature. Concentrated HCl solution (H 2 12.0 M in HO) was added to the mixture to adjust the pH to 4-5. 2 The O was removed in vacuo. The residue was washed with a solution of MeOH:DCM (10%, 30 mL) and filtered. The filtrate was diluted with Na 2 SO 4 Drying at 40° C. and concentration gave crude 3-amino-1-benzylpiperidine-3-carboxylic acid (78-2). LC-MS [M+H] + =235.3.

[0193] 3-Amino-1-benzylpiperidine-3-carboxylic acid (78-2) (5.7 g, 24.33 mmol), K 2 CO 3 (13.45g, 97.31mmol), Boc 2 O (8.2 mL, 36.49 mmol), THF (100 mL), and H 2A mixture of 2,4-dichlorophenyl ether (100 mL) and 1,2,4-tetramethylphenyl ether (1,2,4-tetramethylphenyl ether) was stirred at room temperature for 16 h. The reaction mixture was extracted with EtOAc and the combined organic layers were washed with water, brine, dried over anhydrous sodium sulfate and concentrated to give a residue. The residue was purified by combi-flash (silica gel, KMnO 4 , N.H. 3 H 2 0:MeOH:DCM, 20:100:1000) to give 1-benzyl-3-(tert-butoxycarbonylamino)piperidine-3-carboxylic acid (78-3). LC-MS: [M+H] + =335.3.

[0194] To a solution of 1-benzyl-3-(tert-butoxycarbonylamino)piperidine-3-carboxylic acid (78-3) (670 mg, 2.00 mmol), HATU (1.5 g, 4.00 mmol), and DIPEA (778 mg, 6.00 mmol) in DCM (20 mL) was added CH 3 NH 2 (35% in MeOH, 2 mL) was added. The mixture was stirred at room temperature for 1 h. The reaction mixture was diluted with water, extracted with EtOAc, and the combined organic phase was washed with saturated NaHCO 3 Washing with aqueous solution (20 mL), brine (20 mL), drying over anhydrous sodium sulfate, and concentration gave a residue. The residue was purified by flash chromatography (elution gradient: 10% to 50% EtOAc in PE, 30 min) to give tert-butyl 1-benzyl-3-(methylcarbamoyl)piperidin-3-ylcarbamate (78-4). LC-MS: [M+H] + =348.3.

[0195] To a solution of tert-butyl 1-benzyl-3-(methylcarbamoyl)piperidin-3-ylcarbamate (78-4) (290 mg, 0.836 mmol) in EtOH (10 mL), Pd(OH) 2 (87 mg) was added. The resulting mixture was 2 The mixture was stirred under atmosphere at room temperature for 4 hours. The mixture was filtered, and the filtrate was concentrated to give crude methyl tert-butyl 3-(methylcarbamoyl)piperidin-3-ylcarbamate (Intermediate 78-5).1 H NMR (400MHz, DMSO-d 6 )δ 7.87(s,1H),7.29(s,1H),3.62-3.59(m,1H),3.04-2.87(m,3H),2.61-2.60( m, 3H), 1.83-1.76 (m, 3H), 1.53-1.51 (m, 1H), 1.38 (s, 9H), 1.27-1.25 (m, 1H). LC-MS(UV 214):[M+H] + =258.4.

[0196] Intermediate 89-6: Methyl (3-(2,3-dimethylbutanoyl)piperidin-3-yl)carbamate [ka] To a solution of tert-butyl 3-cyanopiperidine-1-carboxylate (10 g, 47.6 mmol, 1.0 equiv) in dry THF (100 mL) was added 1M LiHMDS (71.4 mL, 71.4 mmol, 1.5 equiv) dropwise under nitrogen at −78° C. The mixture was stirred at −78° C. for 40 min. Then, 3-methylbutanal (6.1 g, 71.4 mmol, 1.5 equiv) was added in one portion at −78° C. The mixture was then stirred at room temperature for 4 h. The mixture was quenched with water (200 mL), extracted with EA (100 mL×2), washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by flash chromatography (PE / EA=92 / 8-90 / 10-70 / 30) to give tert-butyl 3-cyano-3-(1-hydroxy-3-methylbutyl)piperidine-1-carboxylate (89-1). 1 H NMR (400MHz, CD 3OD)δ ppm 4.57(d,J=13.6Hz,0.5H),4.24-3.93(m,1.5H),3.56(dd,J=1.6,10.8Hz,0.5H),3.52-3.42(m,0.5H),3.09-2.56(m,2H),2.22-2.09 (m,0.5H),2.01-1.82(m,1.5H),1.78-1.53(m,4H),1.48(s,9H),1.41-1.29(m,1H),0.99(dd,J=1.6,6.8Hz,3H),0.95-0.89(m,3H). LC-MS:[M+H] + =297.2

[0197] CH 2 Cl 2 To a solution of tert-butyl 3-cyano-3-(1-hydroxy-3-methylbutyl)piperidine-1-carboxylate (89-1) (5.5 g, 18.5 mmol, 1.0 equiv.) in 100 mL of ethyl acetate was added DMP (11.8 g, 27.8 mmol, 1.5 equiv.). The mixture was stirred at room temperature for 30 min. The mixture was diluted with H 2 Dilute with HO (100 mL) and CH 2 Cl 2 (100 mL×2). The organic layer was washed with brine (200 mL), dried over sodium sulfate, filtered, concentrated and purified by combi-flash (PE / EA=80 / 20) to give tert-butyl 3-cyano-3-(3-methylbutanoyl)piperidine-1-carboxylate (89-2). 1 H NMR (400 MHz, CDCl 3 )δ ppm 4.59-3.90(m,2H),3.22-2.92(m,1H),2.87-2.58(m,3H),2.29-2.03(m,2H),1.98-1.69(m,3H),1.50(s,9H),1.03-0.89(m,6H). LC-MS:[M+H] + =295.1.

[0198] To a solution of tert-butyl 3-cyano-3-(3-methylbutanoyl)piperidine-1-carboxylate (89-2) (3.3 g, 11.2 mmol, 1.0 equiv.) in THF (40 mL) was added LiHMDS (12.3 mL, 12.3 mmol, 1.1 equiv., 1 M in THF) and cooled to 30° C. for 1 hour. 2 The mixture was stirred at -78°C for 20 min. Then MeI (1.8 g, 12.3 mmol, 1.1 equiv) was added to the mixture. The mixture was stirred at room temperature for an additional 4 h. The reaction was cooled to 100°C and cooled to 30°C. 2 The mixture was quenched with 2H2O (40 mL) and extracted with EtOAc (40 mL x 2). The organic layers were combined, washed with brine (100 mL), dried over sodium sulfate, filtered, concentrated, and purified by combi-flash (5% EtOAc in hexanes) to give tert-butyl 3-cyano-3-(2,3-dimethylbutanoyl)piperidine-1-carboxylate (89-3). 1 H NMR (400MHz, CD 3 OD)δ ppm 4.50-4.02(m,2H),3.23-2.64(m,3H),2.27-1.72(m,5H),1.49(s,9H),1.16-1.06(m,3H),1.01-0.88(m,6H). LC-MS:[M+H] + =310.2.

[0199] To a mixture of tert-butyl 3-cyano-3-(2,3-dimethylbutanoyl)piperidine-1-carboxylate (89-3) (3.3 g, 11 mmol, 1.0 equiv.) in MeOH (40 mL), NaOH (21 mL, 21 mmol, 1 M / L) and H 2 O 2 (17 mL, 30% aqueous solution) was added. The mixture was stirred at room temperature for 16 h. The mixture was diluted with saturated Na 2 SO 3 The mixture was quenched with aqueous solution (100 mL) and extracted with EtOAc (100 mL×2). The combined organic layers were washed with brine (150 mL) and diluted with anhydrous Na 2 SO 4The residue was purified by silica gel column chromatography (15% to 30% EtOAc in PE) to give tert-butyl 3-carbamoyl-3-(2,3-dimethylbutanoyl)piperidine-1-carboxylate (89-4). 1 H NMR (400 MHz, CDCl 3 )δ ppm 7.15(brs,1H),5.35(brs,1H),4.78-2.28(m,5H),1.94-1.57(m,5H),1.47(s,9H),1.00(dd,J=7.2,20.0Hz,3H),0.90-0.80(m,6H). LC-MS:[M+H] + =327.0.

[0200] A solution of tert-butyl 3-carbamoyl-3-(2,3-dimethylbutanoyl)piperidine-1-carboxylate (89-4) (1.37 g, 4.2 mmol, 1.0 equiv) in MeOH (30 mL) was treated with PhI(OAc) 2 (1.6 g, 5.04 mmol, 1.2 equiv) and KOH (589 mg, 10.5 mmol, 2.5 equiv) were added. The reaction mixture was stirred at 0° C. for 15 h. The mixture was stirred at room temperature for an additional 2 h. The mixture was cooled to 5° C. and cooled to 5° C. for 1 h. 2 The mixture was diluted with 20 mL of 2H2O (40 mL) and extracted with EtOAc (40 mL x 2). The combined organic layers were washed with brine (40 mL) and anhydrous Na 2 SO 4 The residue was purified by silica gel column chromatography (20% EtOAc in PE) to give tert-butyl 3-(2,3-dimethylbutanoyl)-3-((methoxycarbonyl)amino)piperidine-1-carboxylate (89-5). 1 H NMR (400MHz, CD 3 OD):δ 4.74-4.42(m,1H),4.14-3.91(m,1H),3.80-3.48(m,3H),3.29-3.12(m,1 H),2.96-2.64(m,2H),1.98-1.50(m,5H),1.43(s,9H),1.01-0.85(m,9H). LC-MS:[M+H] + =357.1.

[0201] tert-Butyl 3-(2,3-dimethylbutanoyl)-3-((methoxycarbonyl)amino)piperidine-1-carboxylate in DCM (10 mL) To a solution of (89-5) (500 mg, 1.40 mmol, 1.0 equiv) was added TFA (10 mL) at room temperature. The mixture was stirred at room temperature for 30 min. The mixture was concentrated in vacuo and diluted with H 2 Dilute with NH3 H2O (15 mL) and 2 Basify to pH=11 with O and add CHCl 3 The combined organic layer was washed with brine (30 mL) and anhydrous Na 2 SO 4 Drying at 40° C., filtering and concentrating gave methyl (3-(2,3-dimethylbutanoyl)piperidin-3-yl)carbamate (intermediate 89-6). 1 H NMR (400MHz, CD 3 OD)δ ppm 3.63(s,3H),3.27-3.20(m,1H),3.07-2.95(m,1H),2.92-2.81(m,2H),2.66-2.55(m,1H),2.10-2.00( m,1H),1.94-1.76(m,2H),1.72-1.57(m,1H),1.54-1.44(m,1H),1.00-0.95(m,3H),0.91-0.74(m,6H). LC-MS:[M+H] + =257.1.

[0202] Intermediate 113-5: tert-butyl (3-(2,2-difluoro-1-hydroxyethyl)piperidin-3-yl)carbamate [ka] To a solution of 1-benzyl 3-ethyl 3-((tert-butoxycarbonyl)amino)piperidine-1,3-dicarboxylate (5.0 g, 12.30 mmol) in THF (80 mL) and methanol (80 mL) was added lithium chloride (10.43 g, 246 mmol) and sodium tetrahydroborate (9.31 g, 246 mmol) with N2 The reaction mixture was added at room temperature under N 2 The mixture was stirred at room temperature under atmosphere for 18 hours. The reaction mixture was diluted with water and extracted with EtOAc (50 mL×3), and the combined organic phase was washed with water (50 mL), brine (50 mL), dried over anhydrous sodium sulfate, and concentrated to give the crude product. The crude product was purified by flash chromatography (elution gradient: 10% to 50% EtOAc in PE, 30 min) to give benzyl 3-((tert-butoxycarbonyl)amino)-3-(hydroxymethyl)piperidine-1-carboxylate (113-1). LC-MS: [M+H] + =365.2.

[0203] To a solution of benzyl 3-((tert-butoxycarbonyl)amino)-3-(hydroxymethyl)piperidine-1-carboxylate (113-1) (6.2 g, 17.01 mmol) in DCM (50 mL) was added Dess-Martin periodinane (14.43 g, 34.0 mmol) with N 2 The reaction mixture was added at 0 °C under N 2 The mixture was stirred at room temperature under ambient atmosphere for 2 hours. The reaction mixture was diluted with 30 mL of saturated aqueous NaHCO3, extracted with DCM (40 mL x 3), and the combined organic phase was washed with water (50 mL), brine (50 mL), dried over anhydrous sodium sulfate, and concentrated to give the crude product. The crude product was purified by flash chromatography (elution gradient: 10% to 40% EtOAc in PE, 30 min) to give benzyl 3-((tert-butoxycarbonyl)amino)-3-formylpiperidine-1-carboxylate (113-2). LC-MS: [M+H] + =362.1.

[0204] A solution of benzyl 3-((tert-butoxycarbonyl)amino)-3-formylpiperidine-1-carboxylate (113-2) (3 g, 8.28 mmol) and (difluoro(phenylsulfonyl)methyl)trimethylsilane (4.38 g, 16.56 mmol) in anhydrous THF (50 mL) was diluted with a solution of TBAT (0.447 g, 0.828 mmol) in 15 mL of anhydrous THF and N2 After the addition, the reaction mixture was allowed to warm slowly to room temperature and was then cooled to room temperature under N 2 The mixture was stirred at room temperature under atmosphere for 18 hours. Then, TBAF (10.76 mL, 10.76 mmol) was added and the reaction mixture was stirred at room temperature for another 30 minutes. The reaction mixture was diluted with saturated NaCl solution, extracted with EtOAc (50 mL×3), and the combined organic phase was washed with water (40 mL), brine (40 mL), dried over anhydrous sodium sulfate, and concentrated to give the crude product. The crude product was purified by flash chromatography (elution gradient: 10% to 50% EtOAc in hexanes, 40 min) to give benzyl 3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxy-2-(phenylsulfonyl)ethyl)piperidine-1-carboxylate (113-3). LC-MS: [M+H] + =555.2.

[0205] To a solution of benzyl 3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxy-2-(phenylsulfonyl)ethyl)piperidine-1-carboxylate (113-3) (2.4 g, 4.33 mmol) in DMF (50 mL) was added 40 mL of HOAc / NaOAc (1:1) buffer (4 mol / L). Magnesium turnings (1.578 g, 64.9 mmol) were added in portions. The reaction mixture was stirred at room temperature for 4 h. The reaction was quenched with water and extracted with EtOAc (30 mL×3). The combined organic layers were washed with water (30 mL), brine (30 mL) and diluted with anhydrous Na 2 SO 4 The mixture was dried over 1000 ml of ethyl acetate, filtered and concentrated to give a residue. The residue was purified by flash chromatography (elution gradient: 0% to 40% EtOAc in PE, 40 min) to give benzyl, 3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidine-1-carboxylate (113-4). LC-MS: [M+H] + =415.2.

[0206] To a solution of benzyl 3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidine-1-carboxylate (113-4) (1.0 g, 2.413 mmol) in MeOH (50 mL) was added Pd(OH) 2 (0.068 g, 0.483 mmol) was added at room temperature and the reaction mixture was diluted with H 2 Stirred at room temperature under atmosphere for 0.5 hours. The reaction mixture was filtered and concentrated to give tert-butyl (3-(2,2-difluoro-1-hydroxyethyl)piperidin-3-yl)carbamate (Intermediate 113-5). LC-MS: [M+H]+=281.

[0207] Intermediate 156-2: 1-(3-amino-1-(4-(hydroxymethyl)-6-(2,4,5-trifluorophenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol [ka] To a mixture of methyl 5-fluoro-2-(2,4,5-trifluorophenyl)isonicotinate (Intermediate E) (2.5 g, 8.77 mmol) and 1-(3-aminopiperidin-3-yl)-2,2-difluoroethan-1-ol (Intermediate 113-5A) (3.58 g, 8.77 mmol) in DMSO (40 ml) was added DIPEA (40 ml, 229 mmol). The mixture was heated to 100° C. for 3 hours. The mixture was cooled to room temperature and diluted with EtOAc (50 ml) and H 2 The layers were separated and the aqueous layer was extracted with EtOAc (50 mL x 3). The combined organic layers were washed with brine and then with Na 2 SO 4 The mixture was dried at 40° C. and filtered. The filtrate was concentrated. The residue was purified and eluted with EtOAc (0-50%) in n-hexane to give methyl 5-(3-amino-3-(2,2-difluoro-1-hydroxyethyl)piperidin-1-yl)-2-(2,4,5-trifluorophenyl)isonicotinate (intermediate 156-1). 1 H NMR (400 MHz, CDCl 3)δ 8.52(s,1H),8.09-7.98(m,1H),7.92(ddd,J=11.3,9.1,7.2Hz,1H),7.04(td ,J=10.2,6.4Hz,1H),5.92(td,J=55.2,3.5Hz,1H),3.98(s,3H),3.65(td,J= 12.9,3.5Hz,1H),3.36-3.21(m,2H),3.11(d,J=11.8Hz,1H),2.99(ddd,J=12 .2,10.2,3.2Hz,1H),2.02(tdd,J=15.2,9.7,4.6Hz,1H),1.88-1.65(m,3H). LC-MS: [M+H] + =446.1.

[0208] To a mixture of methyl 5-(3-amino-3-(2,2-difluoro-1-hydroxyethyl)piperidin-1-yl)-2-(2,4,5-trifluorophenyl)isonicotinate (Intermediate 156-1) (3.2 g, 7.18 mmol) and LiCl (6.09 g, 144 mmol) in MeOH (100 ml) and THF (100 ml) was added NaBH4 (5.44 g, 144 mmol) at 0° C. The mixture was stirred at room temperature for 4 h. LC-MS showed that most of the SM had been consumed. The mixture was purified by H 2 The mixture was quenched with 2,5-dichloromethane (50 ml) and extracted with EtOAc (50 ml x 4). The combined organic layers were washed with brine and 2 SO 4 The mixture was dried at 40° C. and filtered. The filtrate was concentrated. The filtrate was concentrated and purified by eluting with MeOH in DCM (0-10%) to give 1-(3-amino-1-(4-(hydroxymethyl)-6-(2,4,5-trifluorophenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol (Intermediate 156-2). 1 H NMR (400 MHz, CDCl 3)δ 8.47(s,1H),7.89(ddd,J=11.2,9.1,7.2Hz,1H),7.81(d,J=2.0Hz,1H),7.04(td,J=10.2,6.5Hz,1H),5.92(td,J=55.2,3.6H z,1H),5.12-4.62(m,2H),3.68(td,J=12.5,3.6Hz,1H),3.09(dd,J=25.6,10.6Hz,3H),2.97-2.89(m,1H),2.09-1.72(m,4H). LC-MS:[M+H] + =418.2,419.1.

[0209] Intermediate 158-3: tert-butyl (tert-butoxycarbonyl) (9-((5-(3-(tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidin-1-yl)-2-(2-(difluoromethyl)-4-fluorophenyl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate [ka] To a solution of tert-butyl (3-(2,2-difluoro-1-hydroxyethyl)piperidin-3-yl)carbamate (Intermediate 113-5) (1.828 g, 4.48 mmol) and methyl 2-(2-(difluoromethyl)-4-fluorophenyl)-5-fluoroisonicotinate (Intermediate F) (1.34 g, 4.48 mmol) in DMSO (15 mL), DIPEA (15.64 mL, 90 mmol) was added at room temperature and the reaction mixture was cooled to 5° C. for 2 h. 2 The mixture was stirred under atmospheric pressure at 80° C. for 8 hours. The reaction mixture was diluted with water (40 mL) and extracted with EtOAc (30 mL×3). The combined organic layer was washed with water (20 mL×3), brine (20 mL), and anhydrous Na 2 SO 4The residue was purified by flash chromatography (elution gradient: 0% to 10% MeOH in DCM for 40 min) to give methyl 5-(3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidin-1-yl)-2-(2-(difluoromethyl)-4-fluorophenyl)isonicotinate (Intermediate 158-1). LC-MS: [M+H] + =460.1.

[0210] To a solution of methyl 5-(3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidin-1-yl)-2-(2-(difluoromethyl)-4-fluorophenyl)isonicotinate (Intermediate 158-1) (1.5 g, 3.27 mmol) in THF (30 mL) and MeOH (30 mL) was added NaBH 4 (2.471 g, 65.3 mmol) and LiCl (2.77 g, 65.3 mmol) were added. The reaction mixture was then cooled to 5° C. 2 The mixture was stirred at 20° C. under atmosphere for 2 hours. The reaction mixture was diluted with 30 mL of water and extracted with EtOAc (20 mL×3). The combined organic layers were washed with water (20 mL), brine (20 mL), and concentrated to give the crude product. The crude product was purified by flash chromatography (elution gradient: 0% to 10% MeOH in DCM for 30 min) to give tert-butyl (3-(2,2-difluoro-1-hydroxyethyl)-1-(6-(2-(difluoromethyl)-4-fluorophenyl)-4-(hydroxymethyl)pyridin-3-yl)piperidin-3-yl)carbamate (Intermediate 158-2). LC-MS: [M+H] + =432.

[0211] A mixture of tert-butyl (3-(2,2-difluoro-1-hydroxyethyl)-1-(6-(2-(difluoromethyl)-4-fluorophenyl)-4-(hydroxymethyl)pyridin-3-yl)piperidin-3-yl)carbamate (Intermediate 158-2) (500 mg, 1.159 mmol), Intermediate B (389 mg, 1.159 mmol), and PPh3 (912 mg, 3.48 mmol) was added DEAD (0.550 mL, 3.48 mmol) and the reaction mixture was refluxed with N 2 The mixture was stirred at 0° C. under atmosphere for 30 min. The reaction mixture was diluted with 20 ml of water and extracted with EtOAc (20 mL×3). The combined organic layers were washed with water (20 mL), brine (20 mL) and concentrated to give the crude product. The crude product was purified by flash chromatography (elution gradient: 0% to 10% MeOH in DCM for 30 min) to give tert-butyl (tert-butoxycarbonyl) (9-((5-(3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidin-1-yl)-2-(2-(difluoromethyl)-4-fluorophenyl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate (Intermediate 158-3). LC-MS: [M+H] + =749.

[0212] Intermediate 160-3: tert-butyl (tert-butoxycarbonyl) (9-((5-(3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidin-1-yl)-2-(2,5-difluoro-4-methoxyphenyl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate [ka] To a solution of methyl 2-(2,5-difluoro-4-methoxyphenyl)-5-fluoroisonicotinate (Intermediate G) (2.0 g, 6.73 mmol) and tert-butyl (3-(2,2-difluoro-1-hydroxyethyl)piperidin-3-yl)carbamate (Intermediate 113-5) (2.263 g, 8.07 mmol) in DMSO (10 mL) was added DIPEA (23.50 mL, 135 mmol) and N 2 The mixture was added under atmospheric pressure and stirred at 120° C. for 6 h. The reaction mixture was diluted with water and extracted with EtOAc, and the combined organic phase was washed with water, brine, and sodium chloride. 2 SO 4The crude product was purified by flash chromatography (elution gradient: 10% to 50% EtOAc in hexanes, 30 min) to give methyl 5-(3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidin-1-yl)-2-(2-(difluoromethyl)-4-fluorophenyl)isonicotinate (Intermediate 160-1). LC-MS: [M+H]+=558.1.

[0213] To a solution of methyl 5-(3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidin-1-yl)-2-(2-(difluoromethyl)-4-fluorophenyl)isonicotinate (Intermediate 160-1) (1.9 g, 3.41 mmol) in THF (30 mL) and methanol (30 mL) was added LiCl (0.068 g, 3.41 mmol) and NaBH 4 (0.068 g, 3.41 mmol) was added at 0° C. The reaction mixture was then cooled to 5° C. 2 The mixture was stirred under reduced pressure at room temperature for 2 hours. The reaction mixture was diluted with water, extracted with EtOAc, and the combined organic phase was washed with water, brine, dried over sodium sulfate, and concentrated to give the crude product. The crude product was purified by flash chromatography (elution gradient: 10% to 60% EtOAc in n-hexane, 40 min) to give tert-butyl (3-(2,2-difluoro-1-hydroxyethyl)-1-(6-(2,5-difluoro-4-methoxyphenyl)-4-(hydroxymethyl)pyridin-3-yl)piperidin-3-yl)carbamate (Intermediate 160-2). LC-MS: [M+H] + =530.2.

[0214] A mixture of tert-butyl (3-(2,2-difluoro-1-hydroxyethyl)-1-(6-(2,5-difluoro-4-methoxyphenyl)-4-(hydroxymethyl)pyridin-3-yl)piperidin-3-yl)carbamate (Intermediate 160-2) (1.2 g, 2.3 mmol), Intermediate B (0.76 g, 2.3 mmol), and PPh 3(1.783 g, 6.80 mmol) was dissolved in DEAD (1.1 mL, 6.8 mmol) and 2 The reaction mixture was cooled to 0 °C and cooled to 10 °C. 2 The mixture was stirred under atmospheric pressure for 0.5 h at 0° C. The reaction mixture was quenched with water and extracted with EtOAc, and the combined organic phase was washed with water, brine, and sodium chloride. 2 SO 4 The crude product was purified by flash chromatography (elution gradient: 10% to 50% EtOAc in n-hexane, 50 min) to give tert-butyl (tert-butoxycarbonyl) (9-((5-(3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidin-1-yl)-2-(2,5-difluoro-4-methoxyphenyl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate (Intermediate 160-3). LC-MS: [M+H]+=847.3.

[0215] Following a procedure similar to that for the preparation of tert-butyl (3-(2,2-difluoro-1-hydroxyethyl)-1-(6-(2,5-difluoro-4-methoxyphenyl)-4-(hydroxymethyl)pyridin-3-yl)piperidin-3-yl)carbamate (Intermediate 160-2), the following intermediates (Intermediate Nos. 168-2, 170-2, 172-2) were prepared by reacting the corresponding fluoroisonicotinate with 1-(3-aminopiperidin-3-yl)-2,2-difluoroethan-1-ol (Intermediate 113-5A) (Intermediate Nos. 162-2, 164-2, and 166-2) or tert-butyl (3-(2,2-difluoro-1-hydroxyethyl)piperidin-3-yl)carbamate (Intermediate 113-5). [Table 3-1] [Table 3-2]

[0216] Intermediate 168-3: tert-Butyl (tert-butoxycarbonyl) (9-((5-(3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidin-1-yl)-2-(2-chloro-4-methoxyphenyl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate [ka] A mixture of tert-butyl (1-(6-(2-chloro-4-methoxyphenyl)-4-(hydroxymethyl)pyridin-3-yl)-3-(2,2-difluoro-1-hydroxyethyl)piperidin-3-yl)carbamate (Intermediate 168-2) (0.5 g, 0.947 mmol, 1.0 equiv.), Intermediate B (0.318 g, 0.947 mmol, 1.1 equiv.), and n-PPh 3 (0.745 g, 2.84 mmol, 3 equiv.) was dissolved in DIAD (0.552 mL, 2.84 mmol, 3 equiv.) and N 2 Over a period of 20 min at 0° C. After addition, the reaction mixture was cooled to 5° C. 2 The mixture was stirred at room temperature under atmosphere for 1 h. The reaction mixture was diluted with 20 mL of water and extracted with EtOAc (20 mL×3). The combined organic layers were washed with water (20 mL), brine (20 mL) and concentrated to give the crude product. The crude product was purified by flash chromatography (elution gradient: 20% to 70% EtOAc in n-hexane, 50 min) to give tert-butyl (tert-butoxycarbonyl) (9-((5-(3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidin-1-yl)-2-(2-chloro-4-methoxyphenyl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate (Intermediate 168-3). LC-MS: [M+H] + =845.4.

[0217] Intermediate 170-4: tert-butyl (tert-butoxycarbonyl) (9-((5-(3-(tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidin-1-yl)-2-(2-(difluoromethyl)-4,5-difluorophenyl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate [ka] Step 1. To a solution of tert-butyl (3-(2,2-difluoro-1-hydroxyethyl)-1-(6-(2-(difluoromethyl)-4,5-difluorophenyl)-4-(hydroxymethyl)pyridin-3-yl)piperidin-3-yl)carbamate (Intermediate 170-2) (0.22 g, 0.40 mmol, 1.0 equiv.) in dry DCM (5 mL) was added thionyl chloride (0.146 mL, 2.02 mmol) dropwise at 0° C. The reaction mixture was diluted with saturated K 2 CO 3 The solution was diluted with DCM, washed with water (10 mL), brine (10 mL x 2), and added with Na 2 SO 4 Drying at 40° C. and concentrating under reduced pressure afforded tert-butyl (1-(4-(chloromethyl)-6-(2-(difluoromethyl)-4,5-difluorophenyl)pyridin-3-yl)-3-(2,2-difluoro-1-hydroxyethyl)piperidin-3-yl)carbamate (intermediate 170-3). LC-MS: [M+H] + =568.1.

[0218] Step 2. Mix tert-butyl (1-(4-(chloromethyl)-6-(2-(difluoromethyl)-4,5-difluorophenyl)pyridin-3-yl)-3-(2,2-difluoro-1-hydroxyethyl)piperidin-3-yl)carbamate (Intermediate 170-3) (0.24 g, 0.42 mmol, 1.0 equiv.), Intermediate B (0.14 g, 0.42 mmol), and K in dry DMF (5 mL). 2 CO 3A suspension of (0.175 g, 1.27 mmol, 3.0 equiv) was stirred at room temperature for 16 h and quenched with water. The mixture was extracted with ethyl acetate (10 mL x 3). The combined organic layers were washed with water (10 mL x 3), brine (10 mL) and sodium chloride. 2 SO 4 The crude product was purified by flash chromatography (elution gradient: 10% to 70% EtOAc in hexanes, 50 min) to give tert-butyl (tert-butoxycarbonyl) (9-(5-(3-(tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidin-1-yl)-2-(2-(difluoromethyl)-4,5-difluorophenyl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate (Intermediate 170-4). LC-MS: [M+H] + =867.3.

[0219] Intermediate 172-4: tert-Butyl (tert-butoxycarbonyl) (9-((5-(3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidin-1-yl)-2-(2,3,4-trifluorophenyl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate [ka] Following the same procedure as described in Intermediate 170-4, tert-butyl (3-(2,2-difluoro-1-hydroxyethyl)-1-(4-(hydroxymethyl)-6-(2,3,4-trifluorophenyl)pyridin-3-yl)piperidin-3-yl)carbamate (Intermediate 172-2) was used to obtain tert-butyl (tert-butoxycarbonyl)(9-((5-(3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidin-1-yl)-2-(2,3,4-trifluorophenyl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate (Intermediate 172-4). LC-MS: [M+H] + =835.2.

[0220] Intermediate 174-3: tert-Butyl (tert-butoxycarbonyl) (9-((5-(3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidin-1-yl)-2-(4-fluoro-2-methoxyphenyl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate [ka] A mixture of tert-butyl (3-(2,2-difluoro-1-hydroxyethyl)-1-(6-(4-fluoro-2-methoxyphenyl)-4-(hydroxymethyl)pyridin-3-yl)piperidin-3-yl)carbamate (Intermediate 174-2) (310 mg, 0.61 mmol), Intermediate B (203 mg, 0.61 mmol), and PPh 3 (477 mg, 1.82 mmol), DEAD (0.288 mL, 1.82 mmol) was added dropwise at 0 °C, and the reaction mixture was then cooled to 5 °C. 2 The mixture was stirred at room temperature under atmosphere for 0.5 hours. The reaction mixture was diluted with 20 mL of water and extracted with EtOAc (20 mL×3). The combined organic layers were washed with water (20 mL), brine (20 mL) and concentrated to give the crude product. The crude product was purified by flash chromatography (elution gradient: 10% to 70% EtOAc in PE, 50 min) to give tert-butyl (tert-butoxycarbonyl) (9-((5-(3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidin-1-yl)-2-(4-fluoro-2-methoxyphenyl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate (Intermediate 174-3). LC-MS: [M+H] + =829.2.

[0221] Intermediate 175-3: tert-Butyl (tert-butoxycarbonyl) (9-((5-(3-(tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidin-1-yl)-6'-chloro-5'-fluoro-[2,2-bipyridine]-4-yl)methyl)-9H-purin-6-yl)carbamate [ka] Following the same procedure as intermediate 174-3, tert-butyl (tert-butoxycarbonyl) (9-((5-(3-(tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidin-1-yl)-6'-chloro-5'-fluoro-[2,2'-bipyridine]-4-yl)methyl)-9H-purin-6-yl)carbamate (intermediate 175-3) was obtained. LC-MS: [M+H] + =834.1.

[0222] Intermediate 177-4: Methyl (3-(6-methylpyridin-2-yl)piperidin-3-yl)carbamate hydrochloride

[0223] Intermediate 177-5: Methyl 2-(3,4-difluorophenyl)-5-(3-((methoxycarbonyl)amino)-3-(6-methylpyridin-2-yl)piperidin-1-yl) isonicotinate

[0224] Intermediate 177-6: Methyl (1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridin-3-yl)-3-(6-methylpyridin-2-yl)piperidin-3-yl)carbamate

[0225] Intermediate 177-7: tert-butyl (tert-butoxycarbonyl) (9-((2-(3,4-difluorophenyl)-5-(3-((methoxycarbonyl)amino)-3-(6-methylpyridin-2-yl)piperidin-1-yl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate [ka] To a solution of tert-butyl 3-cyanopiperidine-1-carboxylate (3.00 g, 14.3 mmol, 1.0 equiv.) and 2-fluoro-6-methylpyridine (1.66 g, 14.9 mmol, 1.05 equiv.) in THF (30 mL) was added KHMDS (17 mL, 17 mmol, 1.2 equiv., 1 M in THF) and N 2 The mixture was added dropwise under atmospheric pressure at -70°C. The resulting mixture was warmed to 25°C and stirred for an additional 12 h. The reaction was quenched with water (50 mL) and extracted with EtOAc (30 mL x 3). The combined organic layers were washed with anhydrous Na 2 SO 4 It was dried at 40° C., filtered and concentrated in vacuo. The residue was purified by combi flash (20% EtOAc in PE) to give tert-butyl 3-cyano-3-(6-methylpyridin-2-yl)piperidine-1-carboxylate (177-1). 1 H NMR (400 MHz, CDCl 3 )δ ppm 7.64(t,J=7.6Hz,1H),7.48(d,J=7.6Hz,1H),7.12(d,J=8.0Hz,1H),4.61-4.14(m,2H),3.54-3.21(m,1H),2.94-2 .73(m,1H),2.55(s,3H),2.40-2.24(m,1H),2.23-2.12(m,1H),2.06-1.92(m,1H),1.84-1.75(m,1H),1.51(s,9H). LC-MS:[M+H] + =302.0.

[0226] To a solution of tert-butyl 3-cyano-3-(6-methylpyridin-2-yl)piperidine-1-carboxylate (177-1) (2.70 g, 8.97 mmol, 1.0 equiv) in MeOH (30 mL) was added NaOH (10 mL, 1 M) and H 2 O 2 (5 mL, 30%) was added. The reaction mixture was stirred at 25° C. for 12 h and saturated Na 2 SO 3(100 mL) and stirred at 25° C. for 1 h. The resulting mixture was concentrated in vacuo to remove MeOH. The mixture was extracted with EtOAc (1000 mL×3). The combined organic layers were washed with anhydrous Na 2 SO 4 The mixture was dried at 40° C., filtered and concentrated in vacuo. The residue was purified by silica gel (PE / EtOAc 5 / 1 to 1 / 1) to give tert-butyl 3-carbamoyl-3-(6-methylpyridin-2-yl)piperidine-1-carboxylate (177-2). 1 H NMR (400 MHz, CDCl 3 )δ ppm 7.54(t,J=7.6Hz,1H),7.34(d,J=7.6Hz,1H),7.03(d,J=7.6Hz,1H),5.48(brs,1H),4.67-4.24(m,1H),3.91-3.4 3(m,2H),3.21-2.95(m,1H),2.72-2.55(m,1H),2.53(s,3H),2.32-2.02(m,1H),1.68-1.54(m,2H),1.48(s,9H). LC-MS:[M+H] + =320.1.

[0227] To a solution of tert-butyl 3-carbamoyl-3-(6-methylpyridin-2-yl)piperidine-1-carboxylate (177-2) (2.40 g, 7.51 mmol, 1.0 equiv.) and KOH (1.05 g, 18.8 mmol, 2.5 equiv.) in MeOH (30 mL) was added PhI(OAc) 2 (2.42 g, 7.51 mmol, 1.0 equiv) was added at 0° C. The resulting mixture was stirred at 25° C. for 16 h. The reaction was concentrated in vacuo. The residue was purified by silica gel column (PE / EtOAc 10 / 1 to 3 / 1) to give tert-butyl 3-((methoxycarbonyl)amino)-3-(6-methylpyridin-2-yl)piperidine-1-carboxylate (177-3). 1 H NMR (400 MHz, CDCl 3) δ ppm 7.55 (t, J = 8.0 Hz, 1H), 7.26 (d, J = 8.0 Hz, 1H), 7.01 (d, J = 7.6 Hz, 1H), 4.31 - 3.98 (m, 2H), 3.62 (s, 3H), 3.14 - 2.82 (m, 1H), 2.53 (s, 3H), 2.48 - 2.09 (m, 1H), 1.82 - 1.67 (m, 2H), 1.66 - 1.57 (m, 1H), 1.50 (s, 9H). LC-MS: [M + H] + = 350.3.

[0228] To a mixture of tert-butyl 3-((methoxycarbonyl)amino)-3-(6-methylpyridin-2-yl)piperidine-1-carboxylate (177-3) (1.5 g, 4.30 mmol, 1.0 eq) in DCM (10 mL) was added HCl / dioxane (10 mL, 4 M), and the resulting mixture was stirred at 25 °C for 1 h. The mixture was concentrated in vacuo to afford methyl (3-(6-methylpyridin-2-yl)piperidin-3-yl)carbamate hydrochloride (Intermediate 177-4). 1 H NMR (400 MHz, CD 3 OD) δ ppm 8.48 - 8.39 (m, 1H), 7.92 - 7.78 (m, 2H), 4.33 - 4.25 (m, 1H), 4.09 - 4.01 (m, 1H), 3.68 (s, 3H), 3.50 - 3.39 (m, 1H), 3.30 - 3.17 (m, 1H), 2.91 (s, 3H), 2.44 - 2.31 (m, 1H), 2.30 - 2.14 (m, 2H), 2.06 - 1.94 (m, 1H). LC-MS: [M + H] + = 250.0.

[0229] To a solution of methyl (3-(6-methylpyridin-2-yl)piperidin-3-yl)carbamate hydrochloride (Intermediate 177-4) (1.22 g, 4.27 mmol, 1.0 equiv.) in DMSO (10 mL), DIEA (3.86 g, 29.9 mmol, 7.0 equiv.) and methyl 2-(3,4-difluorophenyl)-5-fluoroisonicotinate (Intermediate A) (1.14 g, 4.27 mmol, 1.0 equiv.) were added. The reaction mixture was stirred at 120° C. for 3 h. The reaction mixture was diluted with water (100 mL) and extracted with EtOAc (20 mL×3). The combined organic layers were washed with brine (20 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography (PE / EA10 / 1-3 / 1) to give methyl 2-(3,4-difluorophenyl)-5-(3-((methoxycarbonyl)amino)-3-(6-methylpyridin-2-yl)piperidin-1-yl)isonicotinate (177-5). 1 H NMR (400 MHz, CDCl 3 )δ ppm 8.52(s,1H),7.94(s,1H),7.87-7.78(m,1H),7.71-7.65(m,1H),7.57(t,J=7.6Hz ,1H),7.35(d,J=8.0Hz,1H),7.26(d,J=7.6Hz,1H),7.02(d,J=7.6Hz,1H),6.75(b rs,1H),4.09(s,3H),3.67(s,3H),3.49-3.34(m,2H),3.12-2.95(m,2H),2.86-2. 74(m,1H),2.54(s,3H),2.40-2.28(m,1H),2.16-1.98(m,1H),1.86-1.81(m,1H). LC-MS:[M+H] + =497.3.

[0230] To a mixture of methyl 2-(3,4-difluorophenyl)-5-(3-((methoxycarbonyl)amino)-3-(6-methylpyridin-2-yl)piperidin-1-yl)isonicotinate (177-5) (600 mg, 1.21 mmol, 1.0 equiv.) and LiCl (512 mg, 12.1 mmol, 10 equiv.) in MeOH / THF (20 mL, v / v=1 / 1) was added NaBH 4(914 mg, 24.2 mmol, 20 equiv.) was added. The reaction mixture was stirred at 25° C. for 1 h. The reaction was quenched with water (50 mL) and extracted with EtOAc (20 mL×3). The combined organic layers were washed with Na 2 SO 4 Drying at 40° C. and concentration in vacuo afforded methyl (1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridin-3-yl)-3-(6-methylpyridin-2-yl)piperidin-3-yl)carbamate (177-6). LC-MS: [M+H] + =469.1.

[0231] A mixture of methyl (1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridin-3-yl)-3-(6-methylpyridin-2-yl)piperidin-3-yl)carbamate (177-6) (500 mg, 1.07 mmol, 1.0 equiv.), intermediate B (537 mg, 1.60 mmol, 1.5 equiv.), and Bu in THF (10 mL). 3 To a solution of P (324 mg, 1.60 mmol, 1.5 equiv) was added DIAD (324 mg, 1.60 mmol, 1.5 equiv). The reaction mixture was stirred at 25 °C for 3 h. The reaction was concentrated in vacuo and the residue was purified by silica gel column chromatography (eluent: PE / EA10 / 1-3:1) to give tert-butyl (tert-butoxycarbonyl) (9-((2-(3,4-difluorophenyl)-5-(3-((methoxycarbonyl)amino)-3-(6-methylpyridin-2-yl)piperidin-1-yl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate (177-7). 1 H NMR (400 MHz, CDCl 3)δ ppm 8.96(s,1H),8.53(s,1H),8.25(s,1H),7.70-7.57(m,2H),7.50-7.43(m,1H),7.36(d,J=8.0Hz,1H) ,7.20-7.11(m,1H),7.06(d,J=8.4Hz,1H),6.51(s,1H),5.84(d,J=15.6Hz,1H),5.45(d,J=15.2Hz, 1H),3.64-3.56(m,1H),3.55(s,3H),3.53-3.46(m,1H),3.17-3.09(m,1H),3.02-2.94(m,1H),2.66 -2.58(m,1H),2.57(s,3H),2.48-2.37(m,1H),2.19-2.05(m,1H),1.94-1.84(m,1H),1.47(s,18H). LC-MS:[M+H] + =786.3.

[0232] Intermediate 269-4: 3-(1-methyl-1H-pyrazol-4-yl)piperidin-3-amine [ka] To a solution of 4-bromo-1H-pyrazole (10.0 g, 68 mmol, 1.0 equiv) in DMF (100 mL) was added NaH (5.44 g, 136 mmol, 1.5 equiv) at 10 °C. After 30 min, MeI (15.0 g, 102 mmol, 2.0 equiv) was added. The mixture was stirred at 30-35 °C for 2 h, quenched with water (200 mL), and extracted with EtOAc (200 mL × 3). The combined organic layers were washed with Na 2 SO 4 and concentrated in vacuo to give 4-bromo-1-methyl-1H-pyrazole (269-1). 1 H NMR (400 MHz, CDCl 3 )δ ppm 7.43(s,1H),7.37(s,1H),3.88(s,3H).

[0233] To a solution of 4-bromo-1-methyl-1H-pyrazole (269-1) (10.6 g, 66.1 mmol, 1.1 equiv.) in THF (140 mL), n-BuLi (29.4 mL, 72.1 mmol, 1.2 equiv., 2.5 M in THF) was added 2 After 30 min, a solution of benzyl 3-oxopiperidine-1-carboxylate (14.0 g, 60.1 mmol, 1.0 equiv) in THF (10 mL) was added. The reaction was stirred at -70°C for 2 h. The reaction was quenched with water (200 mL) and extracted with EtOAc (200 mL x 3). The combined organic layers were washed with brine and diluted with Na 2 SO 4 The mixture was dried at 40° C. and concentrated in vacuo. The residue was purified by combi flash (70-90% EtOAc in PE) to give 3-hydroxy-3-(1-methyl-1H-pyrazol-4-yl)piperidine-1-carboxylate (269-2). LC-MS: [M+H] + =316.1.

[0234] NaN in DCM (30 mL) 3 To a mixture of benzyl 3-hydroxy-3-(1-methyl-1H-pyrazol-4-yl)piperidine-1-carboxylate (Intermediate 269-2) (1.3 g, 19 mmol, 2.0 equiv.) was added TFA (6.00 g, 47.6 mmol, 5.0 equiv.) at 0 °C. After 10 min, a solution of benzyl 3-hydroxy-3-(1-methyl-1H-pyrazol-4-yl)piperidine-1-carboxylate (Intermediate 269-2) (3.00 g, 9.52 mmol, 1.0 equiv.) in DCM (10 mL) was added. The reaction mixture was stirred at 30-35 °C for 16 h. The reaction was chromatographed with NH 3 H 2 The mixture was quenched with O (3 mL), diluted with water (20 mL) and extracted with DCM (300 mL x 3). The combined organic layers were washed with anhydrous Na 2 SO 4 The mixture was dried at 40° C. and concentrated in vacuo. The residue was purified by Combi Flash (40% EtOAc in PE) to give benzyl 3-azido-3-(1-methyl-1H-pyrazol-4-yl)piperidine-1-carboxylate (269-3). LC-MS: [M+H] + =341.1.

[0235] A mixture of benzyl 3-azido-3-(1-methyl-1H-pyrazol-4-yl)piperidine-1-carboxylate (269-3) (1.00 g, 2.9 mmol, 1.0 equiv) and Pd / C (100 mg, 10% wet) in EtOAc (10 mL) was diluted with H 2 The mixture was stirred with a balloon for 16 hours at 30-35° C. The Pd / C was filtered and the filtrate was concentrated in vacuo to give 3-(1-methyl-1H-pyrazol-4-yl)piperidin-3-amine (Intermediate 269-4).

[0236] Intermediate 269-6: Methyl 2-(3,4-difluorophenyl)-5-(3-((methoxycarbonyl)amino)-3-(1-methyl-1H-pyrazol-4-yl)piperidin-1-yl) isonicotinate [ka] Following similar procedures as in the preparation of intermediate 177-5 and the corresponding intermediates, methyl (5-(3-amino-3-(1-methyl-1H-pyrazol-4-yl)piperidin-1-yl)-2-(3,4-difluorophenyl)isonicotinate intermediate 269-5) was prepared. 1 H NMR (400 MHz, CDCl 3 )δ ppm 8.46(s,1H),7.88(s,1H),7.75-7.85(m,1H),7.65-7.70(m,1H),7.53(s,1H),7.50(s,1H),7.15-7.25(m,1 H),3.97(s,3H),3.89(s.3H),3.10-3.30(m,3H),2.90-3.00(m,1H),2.05-2.15(m,1H),1.65-1.90(m,3H).

[0237] Intermediate 269-5 (500 mg, 1.17 mmol, 1.0 equiv) in THF (10 mL) and Et 3To a solution of N (400 mg, 3.51 mmol, 3.0 equiv) was added methyl carbonochloridate (200 mg, 1.75 mmol, 1.5 equiv) at 0 °C. The reaction was stirred at 25-30 °C for 1 h. The reaction was quenched with water (50 mL) and extracted with EtOAc (50 mL × 3). The combined organic layers were washed with Na 2 SO 4 The residue was purified by combi flash (60% EtOAc in PE) to give intermediate 269-6. 1 H NMR (400 MHz, CDCl 3 )δ ppm 8.51(s,1H),7.95(s,1H),7.80-7.90(m,1H),7.60-7.70(m,1H),7.40(s,1H),7.35(s,1H),7.15-7.25(m,1H),6.66(brs,1H),4.03(s .3H),3.85(s,3H),3.64(s,3H),3.30-3.40(m,2H),2.85-3.10(m,2H),2.80(d,J=12.0Hz,1H),1.95-2.05(m,1H),1.60-1.85(m,2H). LC-MS:[M+H] + =486.3.

[0238] Intermediate 269-8: tert-Butyl (tert-butoxycarbonyl) (1-((2-(3,4-difluorophenyl)-5-(3-((methoxycarbonyl)amino)-3-(1-methyl-1H-pyrazol-4-yl)piperidin-1-yl)pyridin-4-yl)methyl)-1H-imidazo[4,5-c]pyridin-4-yl)carbamate [ka] Following the same procedure as in the preparation of intermediate 177-6 and the corresponding intermediates, methyl (1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridin-3-yl)-3-(1-methyl-1H-pyrazol-4-yl)piperidin-3-yl)carbamate (intermediate 269-7) was prepared. LC-MS: [M+H] + =458.2.

[0239] Intermediate 269-8 was prepared using a similar procedure to intermediate 177-7 in example 177, by replacing intermediate 177-6 with intermediate 269-7. LC-MS: [M+H] + =717.4.

[0240] Intermediate 271-7: 3-(pyridin-3-yl)piperidin-3-amine [ka] N 2 To a solution of nicotinaldehyde (17.85 g, 166.7 mmol, 1.0 equiv) in THF (250 mL) at -78 °C under atmospheric pressure was added a solution of 3-butenylmagesium bromide (400 mL, 200 mmol, 0.5 M, 1.2 equiv) dropwise. The reaction mixture was stirred at -78 °C for 2 h, quenched with saturated ammonium chloride solution (300 mL) and separated. The aqueous layer was extracted with ethyl acetate (100 mL x 2). The combined organic layers were washed with brine (200 mL), dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by Combi-flash (30-50% EA in PE) to give 1-(pyridin-3-yl)pent-4-en-1-ol (271-1). 1 HNMR (CDCl 3 400MHz):δ 8.55(d,J=1.6Hz,1H),8.51(dd,J=1.6Hz and 4.8Hz,1H),7.71(dt,J=8.0Hz and 2.0Hz,1H),7.29(dd,J=4.8Hz and 8.0Hz,1H),5.90-5.79(m,1H),5.06(dd,J=1.6Hz and 17.2Hz,1H),5.02(dd,J=1.2Hz and 10.4Hz,1H),4.77(dd,J=5.6Hz and 8.0Hz), 2.34 (brs, 1H), 2.24-2.10 (m, 2H), 1.98-1.88 (m, 1H), 1.86-1.77 (m, 1H).

[0241] CH 2 Cl 2To a solution of 1-(pyridin-3-yl)pent-4-en-1-ol (271-1) (10 g, 61.27 mmol, 1 equiv.) in 10 mL of ethyl acetate (200 mL) was added Dess-Martin periodinane (38.98 g, 91.9 mmol, 1.5 equiv.) in portions at 16-24 °C and the suspension was stirred for 30 min at 16-24 °C. The mixture was poured into saturated sodium bicarbonate solution (300 mL), cooled and stirred for 30 min. The mixture was filtered and the aqueous layer was separated by CH 2 Cl 2 (100 mL×2). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was purified by Combi-flash (20-30% EA in PE) to give 1-(pyridin-3-yl)pent-4-en-1-one (271-2). 1 HNMR (CDCl 3 400MHz):δ 9.18(d,J=1.6Hz,1H),8.78(dd,J=1.6Hz and 4.8Hz,1H),8.23(dt,J=8.0Hz and 2.0Hz,1H),7.43(dd,J=4.8Hz and 8.0Hz, 1H), 5.94-5.84 (m, 1H), 5.12-5.06 (m, 1H), 5.05-5.01 (m, 1H), 2.10 (t, J=7.2Hz, 2H), 2.55-2.48 (m, 2H).

[0242] To a mixture of 1-(pyridin-3-yl)pent-4-en-1-one (271-2) (8.64 g, 53.6 mmol, 1 equiv.) and 2-methylpropane-2-sulfinamide (12.99 g, 107.2 mmol, 2.0 equiv.) in anhydrous THF (120 mL), titanium ethoxide (30.57 g, 134.0 mmol, 2.5 equiv.) was added and the reaction mixture was refluxed at 80° C. for 16 h. The mixture was cooled to room temperature, poured into brine (300 mL), and stirred for 30 min. The mixture was filtered through a celite pad and the aqueous layer was extracted with ethyl acetate (200 mL×2). The combined organic layers were washed with brine (200 mL), dried over anhydrous sodium sulfate, and concentrated. The residue was purified by Combi-flash (20-30% EA in PE) to give (E)-2-methyl-N-(1-(pyridin-3-yl)pent-4-en-1-ylidene)propane-2-sulfinamide (271-3). 1 H NMR (CDCl 3 400MHz):δ 9.05(s,1H),8.70(dd,J=1.6Hz and 4.8Hz,1H),8.10(d,J=7.6Hz,1H),7.38(dd,J=4.8Hz and 8.0Hz, 1H), 5.93-5.81 (m, 1H), 5.09-5.01 (m, 2H), 3.45-3.37 (m, 1H), 3.32-3.25 (m, 1H), 2.55-2.35 (m, 2H).

[0243] To a solution of (E)-2-methyl-N-(1-(pyridin-3-yl)pent-4-en-1-ylidene)propane-2-sulfinamide (271-3) (9.85 g, 37.3 mmol, 1 equiv.) in anhydrous THF (120 mL) was added vinylmagnesium bromide (90 mL, 90 mmol, 2.4 equiv., Et 2A solution of 1M in 2H2O (1M in 2H2O) was added dropwise and the resulting mixture was stirred at -78 °C for 2 h. The reaction was quenched with saturated ammonium chloride (200 mL). The mixture was extracted with ethyl acetate (100 mL x 3) and the combined organic layers were washed with brine (100 ml), dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by Combi-flash (30-70% EA in PE) to give 2-methyl-N-(3-(pyridin-3-yl)hepta-1,6-dien-3-yl)propane-2-sulfinamide (271-4). 1 H NMR (CDCl 3 400MHz):δ 8.89(d,J=1.6Hz,1H),8.53(dd,J=1.6Hz and 4.8Hz,1H),7.82-7.78(m,1H),7.29(dd,J=4.8Hz and 8.0Hz,1H),6.03(dd,J=10.8Hz and 17.2Hz,1H),5.87-5.78(m,1H),5.37(d,J=10.8Hz,1H),5.30(d,J=17.2Hz,1H),5.05-4.96 (m,2H),3.75(s,1H),2.43-2.35(m,1H),2.26-2.18(m,1H),2.07-1.97(m,2H),1.24(s,9H).

[0244] A solution of 2-methyl-N-(3-(pyridin-3-yl)hepta-1,6-dien-3-yl)propane-2-sulfinamide (271-4) (6.90 g, 23.6 mmol, 1.0 equiv.) in methanol (400 mL) at -78 °C was added with O until the solution turned blue. 3 Excess O was bubbled in. 3 N 2The mixture was concentrated in vacuum to remove about 200 mL of methanol. Diphenylmethanilamine (6.48 g, 35.4 mmol, 1.5 equiv), sodium cyanoborohydride (3.72 g, 59.0 mmol, 2.5 equiv), and acetic acid (1.42 g, 23.6 mmol, 1.0 equiv) were added to the above solution and the reaction mixture was stirred at 11-17 °C for 16 h. The mixture was concentrated in vacuum and the residue was diluted with water (100 mL) and extracted with dichloromethane (100 mL x 3). The combined organic layers were concentrated in vacuum and the residue was purified by Combi Flash (30-100% EA in PE) to give N-(1-benzhydryl-3-(pyridin-3-yl)piperidin-3-yl)-2-methylpropane-2-sulfinamide (271-6). 1 H NMR (CDCl 3 400MHz):δ 8.57(d,J=2.0Hz,1H),8.47(dd,J=1.6Hz and 4.8Hz,1H),7.64(dt,J=8.0Hz and 1.6Hz,1H),7.40-7.37(m,4H),7.33-7.28(m,4H),7.25-7.20(m,3H),5.53(brs,1H),4.39(s,1H),3.06-2.99(m,2H) ),2.58(d,J=12.8Hz,1H),2.33-2.25(m,1H),2.09-2.01(m,1H),1.96-1.89(m,2H),1.77-1.73(m,1H),1.28(s,9H). LC-MS: M+H] + =448.2.

[0245] To a solution of N-(1-benzhydryl-3-(pyridin-3-yl)piperidin-3-yl)-2-methylpropane-2-sulfinamide (271-6) (2.34 g, 5.23 mmol, 1.0 equiv) in TFA (24 mL) was added Et 3 SiH (4.68 mL) was added and the reaction mixture was stirred at 80° C. for 16 h. The mixture was concentrated in vacuo, and the residue was diluted with water (50 mL) and extracted with ethyl acetate (30 mL×2) to remove impurities. The aqueous layer was neutralized with sodium hydroxide and concentrated in vacuo to give crude 3-(pyridin-3-yl)piperidin-3-amine (Intermediate 217-7). LC-MS: [M+H]+ =177.9.

[0246] Intermediate 271-9: (5-(3-amino-3-(pyridin-3-yl)piperidin-1-yl)-2-(3,4-difluorophenyl)pyridin-4-yl)methanol [ka] Following similar procedures as in the preparation of intermediate 177-5 and the corresponding intermediates, methyl 5-(3-amino-3-(pyridin-3-yl)piperidin-1-yl)-2-(3,4-difluorophenyl)isonicotinate (intermediate 271-8) was prepared. 1 H NMR (CDCl 3 400MHz):δ 8.90(d,J=2.0Hz,1H),8.54(dd,J=1.6Hz and 4.8Hz,1H),8.46(s,1H),7.98(dt,J=8.0Hz and 1.6Hz,1H),7.88(s,1H),7.85-7.79(m,1H),7.69-7.66(m,1H),7.30(dd,J=4.8Hz and 8.0Hz,1H),7.24-7.19(m,1H),3.97(s,3H),3.34-3.23(m,3H),2.97(td,J=12Hz and 2.4Hz,1H),2.21-2.15(m,1H),2.08(dt,J=12.8Hz and 4.4Hz, 1H), 1.88-1.84 (m, 1H), 1.82-1.76 (m, 1H). LC-MS:[M+H] + =425.2,

[0247] Intermediate 271-9 was prepared using a similar procedure as intermediate 177-6 in example 177, by replacing intermediate 177-5 with intermediate 271-8. 1 H NMR (CDCl 3400MHz):δ 8.81(d,J=2.0Hz,1H),8.52(dd,J=1.6Hz and 4.8Hz,1H),8.41(s,1H),7.88(dt,J=8.0Hz and 1.6Hz,1H),7.85-7.79(m,1H),7.71(s,1H),7.69-7.66(m,1H),7.30(dd,J=4.8Hz and 8.0Hz,1H),7.25-7.18(m,1H),4.92(d,J=14Hz,1H),4.73(d,J=14Hz,1H),3.20(t,J=11.6 Hz,2H),3.09(d,J=11.6Hz,1H),3.01-2.94(m,1H),2.14-2.08(m,2H),1.92-1.87(m,2H). LC-MS:[M+H] + =397.3.

[0248] Intermediate 272-2: 1-Benzyl 3-methyl 2-oxopiperidine-1,3-dicarboxylate [ka] To a solution of piperidin-2-one (10 g, 100 mmol, 1.0 equiv) in dry THF (200 mL) was added NaH (6 g, 60 wt%, 150 mmol, 1.5 equiv) in portions below 0° C. The resulting mixture was stirred at 0° C. for 0.5 h and at 30° C. for 1 h. A solution of benzyl carbonochloridate (26 g, 151.31 mmol, 1.5 equiv) in dry THF (50 mL) was then added to the suspension below 0° C. for 30 min. The resulting mixture was stirred at 30° C. for 3 h. The reaction was quenched with saturated NH 4 The mixture was quenched with Cl (50 mL) and extracted with EtOAc (50 mL x 2). The combined organic layers were washed with brine (50 mL) and diluted with Na 2 SO 4 After filtration and concentration, the residue was purified by silica gel chromatography to give benzyl 2-oxopiperidine-1-carboxylate (272-1). 1 H NMR (CDCl 3,400MHz):δ ppm 7.45-7.32(m,5H),5.28(s,2H),3.78-3.70(m,2H),2.56-2.50(m,2H),1.86-1.80(m,4H).

[0249] To a solution of benzyl 2-oxopiperidine-1-carboxylate (272-1) (9.1 g, 39.1 mmol, 1.0 equiv) in dry THF (200 mL), LDA (30 mL, 2 M, 60 mmol, 1.54 equiv) was added N 2 The mixture was stirred at -70° C. for 30 min under reduced pressure. After stirring at -70° C. for 1 h, methyl carbonochloridate (6.23 g, 65.93 mmol, 1.69 equiv) was added to the mixture over 30 min at below -70° C. The resulting mixture was stirred at -70° C. for 1 h. The reaction mixture was diluted with saturated NH 4 The mixture was quenched with Cl (50 mL) below -70 °C. The mixture was extracted with EtOAc (100 mL x 3). The combined organic layers were washed with brine (100 mL) and dried over Na2SO4. After filtration and concentration, the residue was purified by reverse phase chromatography to give 1-benzyl 3-methyl 2-oxopiperidine-1,3-dicarboxylate (Intermediate 272-2). 1 H NMR (400 MHz, CDCl 3 )δ ppm 7.45-7.32(m,2H),5.29(s,2H),3.80-3.79(d,J=5.2Hz,1H),3.77(s,3H),3.76-3.74(d,J=7.6Hz,1 H),3.58-3.54(dd,J=6.8Hz,J=8.0Hz,1H),2.30-2.16(m,1H),2.14-2.06(m,2H),2.02-1.93(m,1H).

[0250] Intermediate 272-3: (2-Methoxypyridin-3-yl)boronic acid [ka] A suspension of 3-bromo-2-methoxypyridine (9 g, 47.87 mmol, 1.0 equiv), Mg (2.33 g, 95.73 mmol, 2.0 equiv), and LiCl (2.54 g, 59.83 mmol, 1.25 equiv) in dry THF (200 mL) was heated over N 2 The mixture was stirred at 35° C. for 2 hours under reduced pressure. Then, B(OMe) 3 (10 g, 96.23 mmol, 2.01 equiv) was added to the reaction mixture below 0° C. The resulting mixture was then warmed to 35° C. with stirring for 2 h. The reaction was quenched with HCl (0.1 M, 200 mL) to pH=6. The mixture was extracted with EtOAc (100 mL×3). The combined organic layers were washed with brine (100 mL) and diluted with Na 2 SO 4 After filtration and concentration, the residue was washed with chloroform (10 mL) to give (2-methoxypyridin-3-yl)boronic acid (intermediate 272-3). 1 H NMR (400MHz, DMSO-d 6 ) δ ppm 8.19-8.17(dd,J 1 =2.0Hz,J 2 =4.8Hz,1H),7.89-7.88(d,J=2.0Hz,1H),3.98-6.95(dd,J 1 =4.8Hz,J 2 =6.8Hz,1H),3.87(s,3H).

[0251] Intermediate 272-5: 1-benzyl 3-methyl 3-(2-methoxypyridin-3-yl)-2-oxopiperidine-1,3-dicarboxylate [ka] Pb(OAc) in chloroform (20 mL) 4 (6.38 g, 14.39 mmol, 1.0 equiv.) and Hg(OAc) 2 (1.17 g, 3.67 mmol, 0.26 equiv.) was added to a suspension of (2-methoxypyridin-3-yl)boronic acid (272-3) (2.2 g, 14.38 mmol, 1.0 equiv.) in N 2 The suspension was added under N 2The mixture was stirred at 40° C. for 1 h under reduced pressure and at 30° C. for 16 h. The reaction mixture of (2-methoxypyridin-3-yl)plumbanetriyl triacetate (272-4) was used directly in the next step. Then, a solution of 1-benzyl 3-methyl 2-oxopiperidine-1,3-dicarboxylate (272-2) (3.9 g, 13.23 mmol, 0.93 equiv.) in chloroform (20 mL) and pyridine (3 mL) was added to the suspension. The resulting mixture was stirred at 40° C. for 1 h under reduced pressure and at 30° C. for 16 h. The reaction mixture of (2-methoxypyridin-3-yl)plumbanetriyl triacetate (272-4) was used directly in the next step. Then, a solution of 1-benzyl 3-methyl 2-oxopiperidine-1,3-dicarboxylate (272-2) (3.9 g, 13.23 mmol, 0.93 equiv.) in chloroform (20 mL) and pyridine (3 mL) was added to the suspension. 2 The mixture was stirred at 40° C. for 12 hours under reduced pressure. LC-MS showed that intermediate 272-4 was consumed. The reaction mixture was cooled to room temperature and filtered. The filtrate was purified by H 2 SO 4 (1M, 20mL x 4), washed with water (20mL x 2), and then with Na 2 SO 4 After filtration and concentration, the residue was purified by preparative HPLC (FA) to give 1-benzyl 3-methyl 3-(2-methoxypyridin-3-yl)-2-oxopiperidine-1,3-dicarboxylate (intermediate 272-5). 1 H NMR (400 MHz, CDCl 3 )δ ppm 8.12-8.11(m,1H),7.45-7.44(m,2H),7.40-7.31(m,4H),6.88-6.85(m,1H),5.33(s,2H),3.91(s,3H),3.88-3.85( m,1H),3.79(s,3H),3.76-3.69(m,1H),2.66-2.60(m,1H),2.56-2.50(m,1H),1.90-1.82(m,1H),1.77-1.68(m,1H). LC-MS: [M+H] + =398.9.

[0252] Intermediate 272-10: Methyl (3-(2-methoxypyridin-3-yl)piperidin-3-yl)carbamate [ka] To a solution of 1-benzyl 3-methyl 3-(2-methoxypyridin-3-yl)-2-oxopiperidine-1,3-dicarboxylate (Intermediate 272-5) (1.7 g, 4.27 mmol, 1.0 equiv) in dry THF (50 mL) and dry THF (50 mL) was added NaBH 4 (0.3 g, 7.93 mmol, 1.86 equiv.) was added below 0° C. After the addition, the reaction mixture was stirred at 30° C. for 2 h. Then, Pd / C (0.45 g, 0.427 mmol, 0.1 equiv.) was added to the mixture, and the resulting mixture was subjected to H 2 (15 Psi) for 12 h at 30° C. The mixture was filtered and the filtrate was concentrated under reduced pressure to give 3-(2-methoxypyridin-3-yl)piperidine-3-carboxylate (272-6).

[0253] Methyl 3-(2-methoxypyridin-3-yl)piperidine-3-carboxylate (272-6) (2.2 g, 8.79 mmol, 1.0 equiv.) and Boc in MeOH (10 mL). 2 A solution of 272-3H (5.75 g, 26.37 mmol, 3.0 equiv.) was stirred for 16 h at 30° C. The reaction mixture was concentrated and the residue was purified by silica gel column chromatography to give 1-(tert-butyl) 3-methyl 3-(2-methoxypyridin-3-yl)piperidine-1,3-dicarboxylate (272-7). 1 H NMR (400 MHz, CDCl 3 )δ ppm 8.08-8.07(d,J=4.0Hz,1H),7.63-7.61(dd,J 1 =1.6Hz,J 2 =8.6Hz,1H),6.90-6.86(dd,J 1 =5.2Hz,J 2 =7.6Hz,1H),4.15-4.10(m,1H),3.92(s,3H),3.82-3.66(m,2H),3.61(s,3H),3.22(m,1H),2.23-2.19(m,2H),1.71(m,4H),1.32(s,9H). LC-MS:[M+H] + =351.0.

[0254] MeOH (10 mL) and H2 A solution of 1-(tert-butyl) 3-(2-methoxypyridin-3-yl)piperidine-1,3-dicarboxylate (272-7) (0.85 g, 2.43 mmol, 1.0 equiv) and LiOH (0.233 g, 9.70 mmol, 4.0 equiv) in 2H2O (2 mL) was heated to 40 °C with stirring for 2 h and at 80 °C for 4 h. The reaction mixture was concentrated and the residue was acidified with HCl (2N) to pH = 3-4. The mixture was extracted with chloroform / i-PrOH (v / v = 3:1, 10 mL x 4). The combined organic layers were washed with water (10 mL) and concentrated with NaCl. 2 SO 4 The mixture was filtered and the filtrate was concentrated to give 1-(tert-butoxycarbonyl)-3-(2-methoxypyridin-3-yl)piperidine-3-carboxylic acid (272-8). 1 H NMR (400MHz, CD 3 OD)δppm 8.05-8.04(d,J=3.2Hz,1H),7.75-7.73(dd,J 1 =1.6Hz,J 2 =7.2Hz,1H),6.96(s,1H),4.14-4.10(m,1H),4.10-3.89(m,4H),3.62-3.58( m, 1H), 3.30-3.28 (m, 1H), 2.33-2.28 (m, 2H), 1.83-1.79 (m, 2H), 1.27 (s, 9H).

[0255] To a solution of 1-(tert-butoxycarbonyl)-3-(2-methoxypyridin-3-yl)piperidine-3-carboxylic acid (272-8) (0.8 g, 2.38 mmol, 1.0 equiv.) in dry toluene (20 mL) was added DPPA (1.31 g, 4.76 mmol, 2.0 equiv.) and TEA (0.963 g, 9.51 mmol, 4.0 equiv.) in sequence with N 2At 28° C., 100 mL of MeOH was added under reduced pressure. The reaction mixture was stirred at 80° C. for 12 h. Then, MeOH (20 mL) was added to the mixture, and the mixture was stirred at 80° C. for 12 h. The mixture was concentrated under reduced pressure, and the residue was purified by silica gel chromatography to give tert-butyl 3-((methoxycarbonyl)amino)-3-(2-methoxypyridin-3-yl)piperidine-1-carboxylate (272-9). 1 H NMR (400 MHz, CDCl 3 ) δ ppm 8.098.074(dd,J 1 =2.0Hz,J 2 =5.2Hz,1H),7.67-7.64(dd,J 1 =1.6Hz,J 2 =7.6Hz,1H),6.92-6.88(dd,J 1 =4.8Hz,J 2 =7.6Hz,1H),5.78-5.49(m,1H),4.72-4.68(d,13.6Hz,1H),3.99(s,3H),3.53(s,3H),2.98-2.80(m,3H),1.91-1.71(m,4H),1.83(s,9H).

[0256] To a solution of tert-butyl 3-((methoxycarbonyl)amino)-3-(2-methoxypyridin-3-yl)piperidine-1-carboxylate (272-9) (0.6 g, 1.64 mmol, 1.0 equiv) in dry DCM (5 mL) was added TFA (1 mL) below 0° C. After addition, the mixture was allowed to warm to 30° C. with stirring for 3 h. The mixture was concentrated under reduced pressure to give methyl (3-(2-methoxypyridin-3-yl)piperidin-3-yl)carbamate (intermediate 272-10). LC-MS: [M+H] + =266.2.

[0257] Intermediate 272-13: tert-butyl (tert-butoxycarbonyl) (9-((2-(3,4-difluorophenyl)-5-(3-((methoxycarbonyl)amino)-3-(2-methoxypyridin-3-yl)piperidin-1-yl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate [ka] Intermediate 272-13 was prepared following similar procedures as for the preparation of intermediate 177-7 and the corresponding intermediates. 1 H NMR (400 MHz, CDCl 3 )δ ppm 8.93(s,1H),8.50(s,1H),8.27(s,1H),8.14-8.12(dd,J 1 =1.6Hz,J 2 =5.2Hz,1H),7.77-7.75(dd,J 1 =1.6Hz,J 2 =7.6Hz,1H),7.70-7.64(m,1H),7.48-7.46(m,1H),7.20-7.13(dd,J 1 =5.2Hz,J 2 =7.6Hz,1H),6.60(s,1H),5.89-5.85(d,J=15.6Hz,1H),5.43-5.39(d,J=15.6Hz,1H),4.04-4.01(m,1H),3.98(s,3H),3. 49(s,3H),3.36-3.34(m,1H),3.14-3.11(m,1H),2.94-2.84(m,2H),2.21-2.18(m,2H),1.88-1.85(m,1H),1.47(s,18H). LC-MS:[M+H] + =802.5.

[0258] Intermediate 273-5a: tert-butyl 3-cyano-3-(1-(difluoromethyl)-1H-pyrazol-3-yl)piperidine-1-carboxylate

[0259] Intermediate 273-5b: tert-butyl 3-cyano-3-(1-(difluoromethyl)-1H-pyrazol-5-yl)piperidine-1-carboxylate [ka] N 2To a solution of tert-butyl 3-cyanopiperidine-1-carboxylate (25 g, 119.0 mmol, 1.0 equiv) in THF (300 mL) at −70° C. under atmospheric pressure was added LiHMDS (178 mL, 178.57 mmol, 1.5 equiv) dropwise. The reaction mixture was stirred at −70° C. for 30 min and then diluted with CH 3 CHO (7.85 g, 178.57 mmol, 3 equiv.) was added. The reaction was cooled to NH 4 It was quenched with aqueous Cl (200 mL) and extracted with EtOAc (100 mL×3). The combined organic layers were washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by Combi Flash (EtOAc in PE, 10-50%) to give tert-butyl 3-cyano-3-(1-hydroxyethyl)piperidine-1-carboxylate (273-1). 1 H NMR (CDCl 3 400MHz):δppm 4.39-4.05(m,2H),3.75-3.68(m,2H),2.94-2.76(m,2H),2.39-2.20(m,1H),1.94-1.70(m,4H),1.47(s,9H),1.39(t,J=6.4Hz,3H).

[0260] To a solution of tert-butyl 3-cyano-3-(1-hydroxyethyl)piperidine-1-carboxylate (273-1) (9 g, 35.43 mmol, 1.0 equiv.) in DCM (150 mL) was added DMP (22.5 g, 53.15 mmol, 1.5 equiv.). The reaction mixture was stirred at 20° C. for 1 h. The reaction was cooled to 30° C. with NaHCO 3 Aqueous solution (150 mL) and Na 2 SO 3 The mixture was quenched with aqueous NaHCO (150 mL) and extracted with EtOAc (150 mL x 3). The combined organic layers were 3 Washing with aqueous solution (150 mL), drying over anhydrous sodium sulfate, filtering and concentrating gave crude tert-butyl 3-acetyl-3-cyanopiperidine-1-carboxylate (273-2). 1 H NMR (CDCl 3400MHz): δ 4.50-3.85(m,2H),3.20-3.05(m,1H),2.95-2.60(m,1H),2.46(s,3H),2.25-1.70(m,4H),1.48(s,9H).

[0261] To a solution of tert-butyl 3-acetyl-3-cyanopiperidine-1-carboxylate (273-2) (12 g, 47.62 mmol, 1.0 equiv.) in toluene (100 mL) was added DMF-DMA (28.3 g, 238.1 mmol, 5.0 equiv.). The reaction was heated to 100° C. and stirred for 2 h. LC-MS showed a major product. The reaction was concentrated to give tert-butyl (E)-3-cyano-3-(3-(dimethylamino)acryloyl)piperidine-1-carboxylate (273-3). LCMS: [M+H] + =308.0.

[0262] To a mixture of tert-butyl (E)-3-cyano-3-(3-(dimethylamino)acryloyl)piperidine-1-carboxylate (273-3) (5.6 g, 18.22 mmol) in EtOH (30 mL) was added hydrazine hydrate (8.85 mL, 182 mmol), and the mixture was then stirred at 70° C. under Ar atmosphere for 8 h. Water (50 mL) was added, the mixture was extracted with EtOAc, and the combined organic phases were worked up under aqueous conditions and concentrated in vacuo. The residue was purified by flash chromatography (PE / EA, EA: 40%, 30 min) to give tert-butyl 3-cyano-3-(1H-pyrazol-3-yl)piperidine-1-carboxylate (273-4). 1 H NMR (400 MHz, CDCl 3 )δ ppm 7.63(d,J=2.0Hz,1H),6.39(d,J=2.8Hz,1H),4.56-4.46(m,1H),4.25-4.07(m,1H),3.20(d,J=12.0Hz, 1H),2.80-2.90(m,1H),2.37(d,J=13.2Hz,1H),2.10-1.91(m,2H),1.77(d,J=11.0Hz,1H),1.48(s,9H).

[0263] tert-Butyl 3-cyano-3-(1H-pyrazol-3-yl)piperidine-1-carboxylate (273-4) (500 mg, 1.809 mmol) and Cs in DMF (10 mL) 2 CO 3 (2948 mg, 9.05 mmol) was added dropwise with methyl 2-chloro-2,2-difluoroacetate (523 mg, 3.62 mmol) and the reaction mixture was stirred at 60° C. for 24 hours. The reaction mixture was diluted with ammonium chloride solution and the mixture was extracted with ethyl acetate. The organic layer was dried over sodium sulfate, concentrated in vacuo and the residue was purified by flash chromatography (PE / EA, EA: 40%, 30 min) to give tert-butyl 3-cyano-3-(1-(difluoromethyl)-1H-pyrazol-3-yl)piperidine-1-carboxylate (intermediate 273-5a) and tert-butyl 3-cyano-3-(1-(difluoromethyl)-1H-pyrazol-5-yl)piperidine-1-carboxylate (intermediate 273-5b). LC-MS: [M-100] + =227.1.

[0264] Intermediate 273-11: tert-Butyl tert-butoxycarbonyl (9-((5-(3-(1-(difluoromethyl)-1H-pyrazol-3-yl)-3-((methoxycarbonyl)amino)piperidin-1-yl)-2-(4-fluorophenyl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate [ka]

[0265] Intermediate 273-11 was prepared following the same procedure as in the preparation of intermediate 177-7 and the corresponding intermediates. LC-MS: [M+H] + =792.3.

[0266] Intermediate 281-7: Benzyl 3-carbamoyl-3-(2-chlorothiazol-4-yl)piperidine-1-carboxylate [ka] A solution of 1-benzyl 3-methylpiperidine-1,3-dicarboxylate (20 g, 72.12 mmol, 1.0 equiv) in THF (120 mL) was cooled to −78 °C and a solution of LiHMDS (108.18 mL, 108.18 mmol, 1.5 equiv) was added dropwise over 20 min, then stirred at −78 °C for 30 min, followed by CH 3 CHO (3.81 g, 86.54 mmol, 1.2 equiv.) was added. The reaction mixture was stirred at 32° C. for 1 h. The mixture was diluted with saturated NH 4 The mixture was quenched with Cl solution (150 mL) and extracted with EtOAc (200 mL×3). The combined organic phase was washed with brine (400 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by flash column (petroleum ether:EtOAc=9:1-5:1) to give 1-benzyl 3-methyl 3-(1-hydroxyethyl)piperidine-1,3-dicarboxylate (281-1). 1 H NMR (CDCl 3 400MHz):δ 7.36-7.28(m,5H),5.12(s,2H),4.04-3.92(m,1H),3.84-3.77(m,1H),3.66-3.46(m,4H),3.29-3.20(m,1H),3.11 -2.92(m,1H),2.09-1.99(m,1H),1.84-1.79(m,1H),1.70-1.63(m,1H),1.54-1.36(m,1H),1.16(d,J=6.4Hz,3H).

[0267] CH 2 Cl 2 To a mixture of 1-benzyl 3-(1-hydroxyethyl)piperidine-1,3-dicarboxylate (281-1) (17 g, 52.9 mmol, 1.0 equiv.) in 10 mL of 1,2-dichloro-2,4-diphenyl-2,5-diphenyl-1,6-diphenyl-2,7-diphenyl-2,8-diphenyl-2,9-diphenyl-2,7-diphenyl-2,8 ... 3 solution (300 mL), stirred for 30 min until pH was 8, and then added CH 2 Cl 2(300 mL), continued stirring for 30 min, filtered, separated and concentrated the organic phase. The crude product was purified by flash column (petroleum ether: EtOAc = 10: 1) to give 1-benzyl 3-methyl 3-acetylpiperidine-1,3-dicarboxylate (281-2). 1 H NMR (CDCl 3 400MHz): δ.7.34-7.28(m,5H),5.11(s,2H),4.20-3.16(m,7H),2.28-1.84(m,5H),1.76-1.59(m,2H).

[0268] To a solution of 1-benzyl 3-methyl 3-acetylpiperidine-1,3-dicarboxylate (Intermediate 281-2) (15.0 g, 46.9 mmol, 1.0 equiv) in MeOH (200 mL) was added Br 2 (2.6 ml, 51.67 mmol, 1.1 equiv) was added portionwise over 5 min. The reaction mixture was stirred at 60° C. for 1.5 h. The mixture was concentrated and the crude was diluted with EtOAc (300 mL) and saturated Na 2 SO 3 Solution (300 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give 1-benzyl 3-methyl 3-(2-bromoacetyl)piperidine-1,3-dicarboxylate (281-3). LCMS: [M+H] + =399.9.

[0269] To a mixture of 1-benzyl 3-methyl 3-(2-bromoacetyl)piperidine-1,3-dicarboxylate (281-3) (16 g, 32.14 mmol, 1.0 equiv.) and thiourea (4.89 g, 64.28 mmol, 2.0 equiv.) in EtOH (200 mL) was added NaHCO 3(5.40 g, 64.28 mmol, 2.0 equiv) was added. The resulting mixture was heated at 90° C. with stirring for 1.5 h and concentrated in vacuo. The residue was dissolved in EtOAc (300 mL), washed with saturated brine (300 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by flash column (petroleum ether:EtOAc=8:1 to 1:1) to give 1-benzyl 3-methyl 3-(2-aminothiazol-4-yl)piperidine-1,3-dicarboxylate (281-4). 1 H NMR (CDCl 3 400MHz):δ.7.41-7.30(m,5H),6.41-6.23(m,1H),5.53-5.47(m,2H),5.16(s,2H),4.36-4.09(m,2H),3.77-3.68(m,1 H),3.62-3.59(m,3H),3.54-3.48(m,1H),3.37-3.18(m,1H),2.40-2.25(m,2H),2.12-2.01(m,1H),1.61-1.55(m,2H).

[0270] CH 3 To a solution of 1-benzyl 3-methyl 3-(2-aminothiazol-4-yl)piperidine-1,3-dicarboxylate (281-4) (5 g2, 13.32 mmol2, 1.0 equiv) in CN (50 mL) was added CuCl (3.30 g, 33.29 mmol, 2.5 equiv) and heated to 60° C., followed by portionwise addition of t-BuONO (3.43 g, 33.29 mmol, 2.5 equiv) over 5 min. The resulting mixture was then stirred at 80° C. under nitrogen for 1 h. The mixture was cooled to 30° C., diluted with water (50 mL), extracted with EtOAc (60 ml×3), and the organic phase was washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. Two batches of the same scale were performed and combined. The crude product was purified by flash column (petroleum ether:EtoAc=15:1 to 5:1) to give 1-benzyl 3-methyl 3-(2-chlorothiazol-4-yl)piperidine-1,3-dicarboxylate (281-5). 1H NMR(DMSO-d6 400MHz):δ.7.57(s,1H),7.37-7.31(m,5H),5.07(s,2H),4.28-4.13(m,1H),3.69-3.63(m,1H),3.55( s,3H),3.58-3.51(m,1H),3.28-3.18(m,1H),2.32-2.27(m,1H),2.12-2.02(m,1H),1.61-1.42(m,2H).

[0271] MeOH / H 2 To a solution of 1-benzyl 3-methyl 3-(2-chlorothiazol-4-yl)piperidine-1,3-dicarboxylate (281-5) (7.0 g, 17.73 mmol, 1.0 equiv) in 2H2O (80 mL / 10 mL) was added LiOH. 2 2H2O (7.4 g, 177.27 mmol, 10.0 equiv) was added and the resulting mixture was stirred at 30 °C for 2.0 h. The mixture was acidified with 2 N HCl to pH 5-6, diluted with water (40 mL) and extracted with EtOAc (80 mL x 3). The combined organic phase was washed with brine (250 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give 1-((benzyloxy)carbonyl)-3-(2-chlorothiazol-4-yl)piperidine-3-carboxylic acid (281-6). LCMS: [M+Na] + =403.0.

[0272] A solution of 1-((benzyloxy)carbonyl)-3-(2-chlorothiazol-4-yl)piperidine-3-carboxylic acid (281-6) (5.6 g, 14.70 mmol, 1.0 equiv) and NH 4To a solution of Cl (1.57 g, 29.41 mmol, 2.0 equiv.) was added DIEA (7.60 g, 58.82 mmol, 4.0 equiv.) and HATU (11.18 g, 29.41 mmol, 2.0 equiv.). The resulting mixture was stirred at 30° C. for 1 h and the desired MS was observed by LC-MS. The mixture was diluted with water (40 mL) and extracted with EtOAc (80 mL×3). The combined organic phase was washed with brine (200 mL×5), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by flash column (petroleum ether:EtOAc=5:1 to 1:1) to give benzyl 3-carbamoyl-3-(2-chlorothiazol-4-yl)piperidine-1-carboxylate (intermediate 281-7). 1 H NMR (DMSO-d 6 400MHz):δ.7.44-7.31(m,6H),7.13(s,1H),7.01(s,1H),5.06(s,2H),4.07-4.03(m,1H),3.78-3.63(m,1H),3.58-3.46(m,1H),3.18(br s,1H),2.33-2.25(m,1H),2.13-2.08(m,1H),1.61-1.46(m,1H),1.40-1.29(m,1H).

[0273] Intermediate 281-10: Methyl 5-(3-(2-chlorothiazol-4-yl)-3-((methoxycarbonyl)amino)piperidin-1-yl)-2-(4-fluorophenyl)isonicotinate [ka] Following similar procedures as in the preparation of intermediate 177-3 and the corresponding intermediates, 3-(2-chlorothiazol-4-yl)-3-((methoxycarbonyl)amino)piperidine-1-carboxylate (intermediate 281-8) was prepared. 1 H NMR (CDCl 3400MHz):δ.7.37-7.30(m,5H),7.18-7.07(m,1H),5.44-5.11(m,3H),4.23-4.08(m,1H),3.92-3.89(m,1H),3.74- 3.56(m,4H),3.14(t,J=10.4Hz,1H),2.71-2.50(m,1H),2.25-2.12(m,1H),1.71-1.65(m,1H),1.60-1.49(m,1H).

[0274] A solution of benzyl 3-(2-chlorothiazol-4-yl)-3-((methoxycarbonyl)amino)piperidine-1-carboxylate (Intermediate 281-8) (1.5 g, 3.66 mmol, 1.0 equiv) in TFA (12.0 mL) was stirred at 66° C. for 4.0 h. The mixture was concentrated in vacuo and the residue was dissolved in EtOAc (20 mL) and saturated NaHCO 3 The solution was washed with (40 mL×3), dried over anhydrous sodium sulfate, filtered and concentrated to give methyl (3-(2-chlorothiazol-4-yl)piperidin-3-yl)carbamate (281-9).

[0275] To a solution of methyl (3-(2-chlorothiazol-4-yl)piperidin-3-yl)carbamate (281-9) (1.3 g, 3.63 mmol, 1.0 equiv) and intermediate C (903.7 mg, 3.63 mmol, 1.0 equiv) in DMSO (20 mL) was added DIEPA (1.87 g, 14.51 mmol, 4.0 equiv). The reaction mixture was stirred at 120° C. for 12 h. The desired MS was observed by LCMS. The mixture was cooled to 30° C., diluted with water (20 mL), and extracted with EtOAc (30 mL×3). The organic phase was washed with brine (60 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by flash column (petroleum ether: EtOAc = 15: 1 to 6: 1) to give methyl 5- (3- (2-chlorothiazol-4-yl) -3- ( (methoxycarbonyl) amino) piperidin-1-yl) -2- (4-fluorophenyl) isonicotinate (intermediate 281-10). 1 H NMR (CDCl 3400MHz):δ.8.53(s,1H),7.97(s,1H),7.96-7.92(m,2H),7.18-7.12(m,3H),6.82(brs,1H),4.04(s,3H),3.66(s,3H),3. 50(d,J=12.4Hz,1H),3.32(d,J=10.0Hz,1H),3.10-2.98(m,2H),2.93-2.84(m,1H),2.06-1.95(m,2H),1.87-1.76(m,1H).

[0276] Intermediate 281-12: tert-butyl (tert-butoxycarbonyl) (9-((5-(3-(2-chlorothiazol-4-yl)-3-((methoxycarbonyl)amino)piperidin-1-yl)-2-(4-fluorophenyl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate [ka] Following a procedure similar to that for the preparation of intermediate 177-6, methyl (3-(2-chlorothiazol-4-yl)-1-(6-(4-fluorophenyl)-4-(hydroxymethyl)pyridin-3-yl)piperidin-3-yl)carbamate (intermediate 281-11) was prepared. 1 H NMR (CDCl 3 400MHz):δ.8.45(s,1H),7.94-7.90(m,2H),7.64(s,1H),7.15-7.11(m,3H ),7.15-7.10(m,2H),5.75(brs,1H),4.89-4.78(m,2H),3.63(s,3H),3.60 -3.56(m,1H),3.30-3.26(m,1H),3.15-3.11(m,1H),2.99-2.92(m,1H),2. 60-2.57(m,1H),2.14-2.07(m,1H),2.00-1.93(m,1H),1.85-1.79(m,1H).

[0277] Intermediate 281-12 was prepared following the same procedure as for the preparation of intermediate 177-7 and the corresponding intermediate. LCMS: [M+H] + =794.4

[0278] Intermediate 282-4: 1-Benzyl 3-methyl 3-(2H-1,2,3-triazol-4-yl)piperidine-1,3-dicarboxylate [ka] To a solution of 1-benzyl 3-methylpiperidine-1,3-dicarboxylate (30 g, 108 mmol, 1.0 equiv.) in THF (350 mL), LiHMDS (163 mL, 163 mmol, 1.5 equiv.) was added N 2 The mixture was added dropwise under atmospheric pressure at -65°C. After stirring for 30 min, paraformaldehyde (9.72 g, 324 mmol, 3.0 equiv) was added. The reaction mixture was stirred at 13-23°C for 1 h. The reaction was quenched with water (300 mL), extracted with EtOAc (300 mL x 3), and washed with brine (500 mL x 2). The combined organic layers were washed with Na 2 SO 4 The residue was purified by CombiFlash (20-40% EtOAc in PE) to give 1-benzyl 3-methyl 3-(hydroxymethyl)piperidine-1,3-dicarboxylate (282-1). 1 H NMR (400 MHz, CDCl 3 )δppm 7.40-7.26(m,5H),5.15(s,2H),4.06-3.80(m,1H),3.75-3.60(m,6H),3.50-3.25( m,1H),3.15-2.85(m,1H),1.84-1.71(m,1H),1.70-1.62(m,1H),1.61-1.50(m,2H). LC-MS:[M+H] + =308.0.

[0279] To a solution of 1-benzyl 3-methyl 3-(hydroxymethyl)piperidine-1,3-dicarboxylate (282-1) (13.4 g, 43.6 mmol, 1.0 equiv.) in dry DCM (150 mL) was added DMP (36.97 g, 87.21 mmol, 2.0 equiv.) in portions at 0 °C. The resulting mixture was stirred at 11-20 °C for 2 h. The mixture was diluted with saturated NaHCO 3Pour into solution (300 mL), extract with DCM (300 mL x 3), and add NaHCO 3 Wash with aqueous solution (400 mL x 2) and add Na 2 SO 4 The residue was purified by flash chromatography (PE / EA=80 / 20) to give 1-benzyl 3-methyl 3-formylpiperidine-1,3-dicarboxylate (282-2). 1 H NMR (400 MHz, CDCl 3 )δ ppm 9.69-9.53(m,1H),7.43-7.28(m,5H),5.14(s,2H),4.42-4.04(m,1H),3. 86-3.55(m,5H),3.35-3.11(m,1H),2.28-1.90(m,2H),1.73-1.54(m,2H).

[0280] To a solution of 1-benzyl 3-methyl 3-formylpiperidine-1,3-carboxylate (282-2) (6.5 g, 21.29 mmol, 1.0 equiv.) and compdimethyl(1-diazo-2-oxopropyl)phosphonate (8.17 g, 42.58 mmol, 2.0 equiv.) in dry methanol (120 mL) was added K 2 CO 3 (8.8 g, 63.87 mmol, 3.0 equiv) was added. The reaction mixture was stirred at 15-22 °C for 16 h. The mixture was concentrated. The residue was purified by flash chromatography (PE / EA = 60 / 40) to give 1-benzyl 3-methyl 3-ethynylpiperidine-1,3-dicarboxylate (282-3). 1 H NMR (400 MHz, CDCl 3 )δ ppm 7.41-7.29(m,5H),5.15(s,2H),4.21-4.07(m,1H),4.00-3.82(m,1H),3.75(s,3H),3.54-3.41(m,1 H),3.14-2.99(m,1H),2.26-2.17(m,1H),2.10-1.82(m,3H),1.92-1.83(m,1H),1.68-1.58(m,1H).

[0281] 1-Benzyl 3-methyl in DMF / MeOH (40 mL / 4 mL) 3 To a solution of -ethynylpiperidine-1,3-dicarboxylate (Intermediate 282-3) (4 g, 13.27 mmol, 1.0 equiv.), TMSN 3 (2.3 g, 19.91 mmol, 1.5 equiv.) was added. The mixture was stirred at 100-110 °C for 16 h, diluted with water (100 mL), extracted with EA (100 mL × 4), washed with brine (100 mL × 3), and concentrated in 10 mL of NaCl. 2 SO 4 Drying at 40° C., filtration and concentration gave the crude compound, which was purified by CombiFlash (DCM / MeOH=10 / 1) to give 1-benzyl 3-methyl 3-(2H-1,2,3-triazol-4-yl)piperidine-1,3-dicarboxylate (Intermediate 282-4). LC-MS: [M+H] + =345.0.

[0282] Intermediate 282-5a: 1-benzyl 3-methyl 3-(2-methyl-2H-1,2,3-triazol-4-yl)piperidine-1,3-dicarboxylate

[0283] Intermediate 282-5b: 1-benzyl 3-methyl 3-(1-methyl-1H-1,2,3-triazol-4-yl)piperidine-1,3-dicarboxylate [ka] To a solution of 1-benzyl 3-methyl 3-(2H-1,2,3-triazol-4-yl)piperidine-1,3-dicarboxylate (Intermediate 282-4) (3.3 g, 9.58 mmol, 1.0 equiv.) in DMF (40 mL) was added Cs 2 CO 3(15.6 g, 47.9 mmol, 5 eq.) and MeI (9.3 g, 28.74 mmol, 3 eq.) were added. The reaction mixture was stirred at 13-18 °C for 16 h. The mixture was filtered and the filtrate was concentrated to give crude compounds which were purified by preparative HPLC to give 1-benzyl 3-methyl 3-(2-methyl-2H-1,2,3-triazol-4-yl)piperidine-1,3-dicarboxylate (Intermediate 282-5a) and 1-benzyl 3-methyl 3-(1-methyl-1H-1,2,3-triazol-4-yl)piperidine-1,3-dicarboxylate (Intermediate 282-5b).

[0284] Intermediate 282-5a: 1 H NMR (400MHz, CD 3 OD)δ ppm 7.63-7.47(m,1H),7.44-7.27(m,5H),5.15-5.07(m,2H),4.40-4.15(m,1H),4.09(s,3H),3.90-3. 70(m,1H),3.60(brs,4H),3.48-3.32(m,1H),2.38(brs,1H),2.26-2.10(m,1H),1.70-1.55(m,2H). LC-MS:[M+H] + =359.3.

[0285] Intermediate 282-5b: 1 H NMR (400MHz, CD 3 OD)δ ppm 7.81(s,1H),7.44-7.27(m,5H),5.12(s,2H),4.48-4.04(m,1H),4.00(s,3H),3.94-3.82(m,0.6 H),3.73-3.43(m,2H),3.60(s,3H),3.28-3.17(m,0.4H),2.48-2.10(m,2H),1.82-1.47(m,2H).

[0286] Intermediate 282-6: 1-((benzyloxy)carbonyl)-3-(1-methyl-1H-1,2,3-triazol-4-yl)piperidine-3-carboxylate [ka] Intermediate 282-6 was prepared using a similar procedure to intermediate 281-6 in example 281, by replacing intermediate 281-5 with intermediate 282-5a. LC-MS: [M+H] + =345.4.

[0287] Intermediate 282-7: Benzyl 3-carbamoyl-3-(1-methyl-1H-1,2,3-triazol-4-yl)piperidine-1-carboxylate [ka] Intermediate 282-7 was prepared using a similar procedure to intermediate 281-7 in example 281, by replacing intermediate 281-6 with intermediate 282-6. LC-MS: [M+H] + =344.1.

[0288] Intermediate 282-8: benzyl 3-((methoxycarbonyl)amino)-3-(1-methyl-1H-1,2,3-triazol-4-yl)piperidine-1-carboxylate [ka] Intermediate 282-8 was prepared using a similar procedure to intermediate 177-3 in example 177, by replacing intermediate 177-3 with intermediate 281-7. 1 H NMR (400 MHz, CDCl 3 )δ ppm 7.64-7.30(m,6H),5.29-5.10(m,2H),4.09-3.71(m,6H),3.57(s,3H),3.30(brs, 1H), 2.69-2.45 (m, 1H), 2.37-2.15 (m, 1H), 1.82-1.65 (m, 1H), 1.55-1.45 (m, 1H). LC-MS:[M+H] + =374.1.

[0289] Intermediate 282-9: Methyl (3-(1-methyl-1H-1,2,3-triazol-4-yl)piperidin-3-yl)carbamate [ka] Intermediate 282-9 was prepared using a similar procedure as intermediate 269-4 in example 269, by replacing intermediate 269-3 with intermediate 281-8. 1 H NMR (400MHz, CD 3 OD)δ ppm 7.81(s,1H),4.06(s,3H),3.56(s,3H),3.40-3.32(m,1H),3.13-2.89(m,2H),2.71-2.62 (m,1H),2.39(d,J=13.6Hz,1H),2.08-1.99(m,1H),1.79-1.66(m,1H),1.57-1.47(m,1H).

[0290] Intermediate 282-10: Methyl 2-(3,4-difluorophenyl)-5-(3-((methoxycarbonyl)amino)-3-(1-methyl-1H-1,2,3-triazol-4-yl)piperidin-1-yl)isonicotinate [ka] Intermediate 282-10 was prepared using a similar procedure as intermediate 177-5 in example 177, by replacing intermediate 177-4 with intermediate 282-9. 1 H NMR (400 MHz, CDCl 3 )δ ppm 8.56(s,1H),7.98(s,1H),7.87-7.81(m,12Hz,1H),7.72-7.67(m,1H),7.64(s,1) H),7.27-7.20(m,1H),6.87(brs,1H),4.07(s,3H),4.03(s,3H),3.72-3.68(m,0. 5H),3.66(s,3H),3.60-3.50(m,0.5H),3.35(d,J=11.2Hz,1H),3.18(d,J=12.4Hz ,1H),3.10(td,J=11.6Hz,2.8Hz,1H),2.94(d,J=11.2Hz,1H),2.09-1.70(m,3H). LC-MS:[M+H] + =487.5.

[0291] Intermediate 282-11: Methyl (1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridin-3-yl)-3-(1-methyl-1H-1,2,3-triazol-4-yl)piperidin-3-yl)carbamate [ka] Intermediate 282-11 was prepared using a similar procedure as intermediate 177-6 in example 177, by replacing intermediate 177-5 with intermediate 282-10. 1 H NMR (400 MHz, CDCl 3 )δ ppm 8.46(s,1H),7.86-7.80(m,1H),7.70-7.66(m,1H),7.65(s,1H),7.57(s,1H),7.25- 7.19(m,1H),5.75(brs,1H),4.85-4.77(m,2H),4.08(s,3H),3.95(brs,1H),3.62(s ,3H),3.69-3.52(m,1H),3.42(d,J=11.2Hz,1H),3.13(d,J=11.6Hz,1H),3.03-2.94 (m,1H),2.64-2.52(m,1H),2.25-2.10(m,1H),2.02-1.94(m,1H),1.85-1.77(m,1H). LC-MS:[M+H] + =459.1.

[0292] Intermediate 282-12: tert-Butyl (tert-butoxycarbonyl) (9-((2-(3,4-difluorophenyl)-5-(3-((methoxycarbonyl)amino)-3-(1-methyl-1H-1,2,3-triazol-4-yl)piperidin-1-yl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate [ka] Intermediate 282-12 was prepared using a similar procedure to intermediate 177-7 in example 177, by replacing intermediate 177-6 with intermediate 282-11. LC-MS: [M+H] + =776.3.

[0293] Intermediate: 284-4: 3-(5-methylisoxazol-3-yl)piperidin-3-amine [ka] EtOH / H 2 Benzyl 3-((tert-butoxycarbonyl)amino)-3-formylpiperidine-1-carboxylate (238 mg, 0.656 mmol) and hydroxylamine HCl (38 mg, 0.547 mmol) in 2O (1:1, 20 mL) and K 2 CO 3 (91 mg, 0.656 mmol) was stirred at room temperature overnight and then added EA (20 mL) and H 2 The layers were separated and the aqueous layer was extracted with EA (10 mL×2). The combined organic layers were washed with brine and then with Na 2 SO 4 The mixture was dried over 1000 ml and concentrated. The filtrate was concentrated to give benzyl (E)-3-((tert-butoxycarbonyl)amino)-3-((hydroxyimino)methyl)piperidine-1-carboxylate (284-1). LC-MS: [M+H-100] + =277.9.

[0294] To a mixture of benzyl (E)-3-((tert-butoxycarbonyl)amino)-3-((hydroxyimino)methyl)piperidine-1-carboxylate (284-1) (2 g, 5.3 mmol) in DMF (20 mL) was added NCS (1.061 g, 7.95 mmol). The mixture was stirred at 40° C. for 2 h. The mixture was then diluted with EA (20 mL) and H 2 The layers were separated and the aqueous layer was extracted with EA (20 mL×2). The combined organic layers were washed with brine and 2 SO 4 The filtrate was concentrated to give crude benzyl (Z)-3-((tert-butoxycarbonyl)amino)-3-(chloro(hydroxyimino)methyl)piperidine-1-carboxylate (284-2).

[0295] To a mixture of crude benzyl (Z)-3-((tert-butoxycarbonyl)amino)-3-(chloro(hydroxyimino)methyl)piperidine-1-carboxylate (284-2) (2 g, 4.86 mmol) in DCM (50 mL) was added 2-chloroprop-1-ene (7.43 g, 97 mmol) and TEA (3.41 mL, 24.28 mmol). The mixture was heated to 40° C. overnight. Then the mixture was concentrated. The residue was purified and eluted with EA in n-hexane (0-50%) to give benzyl 3-((tert-butoxycarbonyl)amino)-3-(5-methylisoxazol-3-yl)piperidine-1-carboxylate (284-3). LC-MS: [M+H] + =415.9.

[0296] A mixture of benzyl 3-((tert-butoxycarbonyl)amino)-3-(5-methylisoxazol-3-yl)piperidine-1-carboxylate (284-3) (700 mg, 1.685 mmol) in HBr in HOAc (10 mL) was stirred at room temperature for 8 h. The mixture was concentrated and the residue was adjusted to pH=10-12 with an aqueous solution of 1 M NaOH. The mixture was then extracted with i-PrOH / DCM (1:3 20 mL×3). The combined organic layers were washed with Na 2 SO 4 The mixture was dried at 40° C. and filtered. The filtrate was concentrated to give 3-(5-methylisoxazol-3-yl)piperidin-3-amine (Intermediate 284-4). LC-MS: [M+H] + =182.0.

[0297] Intermediate 284-6: (5-(3-amino-3-(5-methylisoxazol-3-yl)piperidin-1-yl)-2-(3,4-difluorophenyl)pyridin-4-yl)methanol [ka] Methyl 5-(3-amino-3-(5-methylisoxazol-3-yl)piperidin-1-yl)-2-(3,4-difluorophenyl)isonicotinate (intermediate 284-5) was prepared from the corresponding intermediate following a procedure similar to that described in Example 177-5. LC-MS: [M+H] + =428.9,429.8.

[0298] Intermediate 284-6 was prepared using a similar procedure to intermediate 177-6 in example 177, by replacing intermediate 177-5 with intermediate 284-5. LC-MS: [M+H] + =400.9,401.9.

[0299] Intermediate 285-4: 3-(2-(benzyloxy)-1-fluoroethyl)piperidin-3-amine [ka] To a solution of 2-(benzyloxy)acetaldehyde (0.652 g, 4.34 mmol) in 0.025 M aqueous NaOH (15 mL, 0.375 mmol), 2-(benzyloxy)acetaldehyde (0.652 g, 4.34 mmol) and N,N,N-trimethylhexadecan-1-aminium chloride (0.139 g, 0.434 mmol) were added at room temperature, and the mixture was cooled to 10° C. for 2 h. 2 The mixture was stirred at room temperature under atmospheric pressure for 3 hours. The reaction mixture was diluted with water and extracted with EtOAc (20 ml×3), and the combined organic phase was washed with water (20 ml), brine (20 ml), dried over anhydrous sodium sulfate, and concentrated in vacuo to give a residue. The residue was purified by flash chromatography (elution gradient: 0% to 30% EtOAc in PE, 30 min) to give tert-butyl 3-(2-(benzyloxy)-1-hydroxyethyl)-3-nitropiperidine-1-carboxylate (285-1). 1 H NMR (400MHz, DMSO-d 6)δ ppm.40-7.24(m,6H),5.84(dd,J=28.5,6.2Hz,1H),4.79-4.56(m,1H),4.54-4.44(m ,2H),4.07-3.91(m,1H),3.85(s,1H),3.59(ddd,J=27.8,10.5,4.9Hz,1H),3.39(dd d,J=12.0,10.4,6.3Hz,1H),3.18(dd,J=22.8,13.2Hz,1H),2.72(s,1H),2.41(t,J= 15.6Hz, 1H), 1.95-1.77 (m, 1H), 1.61 (dt, J = 14.0, 3.9Hz, 1H), 1.37 (d, J = 2.5Hz, 9H). LC-MS:[M+H-100] + =281.2.

[0300] To a solution of tert-butyl 3-(2-(benzyloxy)-1-hydroxyethyl)-3-nitropiperidine-1-carboxylate (285-1) (0.93 g, 2.445 mmol) in DCM (20 mL) was added DAST (0.646 mL, 4.89 mmol) at 0° C. and the mixture was cooled to 5° C. with N 2 The mixture was stirred at 0° C. under ambient atmosphere for 3 h. The reaction mixture was then diluted with water, extracted with EtOAc (20 mL×3), and worked up in aqueous conditions to give a residue that when concentrated in vacuo was purified by flash chromatography (elution gradient: 0% to 40% EtOAc in PE, 30 min) to give tert-butyl 3-(2-(benzyloxy)-1-fluoroethyl)-3-nitropiperidine-1-carboxylate (285-2). LC-MS: [M+H-100]+=383.1.

[0301] To a solution of tert-butyl 3-(2-(benzyloxy)-1-fluoroethyl)-3-nitropiperidine-1-carboxylate (285-2) (200 mg, 0.684 mmol) in EtOH (10 mL) was added Zn powder (44.7 mg, 0.684 mmol) at room temperature, and the mixture was then purified by N 2After stirring at 0° C. for 3 h under ambient conditions, 4M HCl (2.281 mL, 6.84 mmol) was added dropwise at 0° C. The reaction mixture was filtered and concentrated to give a residue. The residue was purified by flash chromatography (elution gradient: 0% to 40% EtOAc in PE, 30 min) to give tert-butyl 3-amino-3-(2-(benzyloxy)-1-fluoroethyl)piperidine-1-carboxylate (285-3). LC-MS: [M+H] + =263.2.

[0302] To a solution of tert-butyl 3-amino-3-(2-(benzyloxy)-1-fluoroethyl)piperidine-1-carboxylate (285-3) (180 mg, 0.511 mmol) in DCM (15 mL) was added TFA (5 mL, 64.9 mmol) at room temperature and the mixture was purified by N 2 Stirred at room temperature under atmosphere for 1 hour.The mixture was then concentrated in vacuo to give 3-(2-(benzyloxy)-1-fluoroethyl)piperidin-3-amine (Intermediate 285-4) as a TFA salt.LC-MS:[M+H] + =253.2.

[0303] Intermediate 287-6: 2,2,2-trifluoroacetaldehyde compound and N-benzyl-3-(1,2-difluoroethyl)piperidin-3-amine (1:1) [ka] AlLiH in THF (200 mL) 4 (13.42 g, 353.45 mmol) was stirred at 80° C. for 2 h. The mixture was then added to ethyl 2-fluoroacetate (150 g, 1.41 mol) in THF (100 mL) at −60° C. and stirred at −60° C. for 1 h. The mixture was quenched with EtOH (20 mL) at −60° C. and washed with ice water (500 mL) and H 2 SO 4(50 mL) and extracted with MTBE (150 mL x 3). The organic layer was concentrated in vacuum (25 °C) to give a residue. The residue was purified by distillation (-0.095 MPa, 25-50 °C) to give 1-ethoxy-2-fluoroethan-1-ol (287-1). 1 H NMR (400MHz, CD 3 OD)δ=4.30-4.16(m,2H),3.61(q,J=7.1Hz,2H),1.20-1.18(m,3H).

[0304] To a solution of tert-butyl 3-nitropiperidine-1-carboxylate (40 g, 173.72 mmol) in THF (40 mL), 1-ethoxy-2-fluoroethan-1-ol (287-1) (93.90 g, 868.58 mmol), K 2 CO 3 (12.00 g, 86.86 mmol) was added and stirred at 40° C. for 4 hours. The mixture was concentrated in vacuo to give a residue. The residue was purified by column chromatography (PE / EA=50:1 to 10:1) to give tert-butyl 3-(2-fluoro-1-hydroxyethyl)-3-nitropiperidine-1-carboxylate (287-2). 1 H NMR (400 MHz, CDCl 3 )δ=4.78-4.62(m,1H),4.60-4.50(m,1H),4.50-4.41(m,1H),4.22-4.02(m,1H),3.99-3.60(m, 1H),3.48-2.92(m,2H),2.57-2.37(m,1H),2.34-2.10(m,1H),1.74-1.64(m,2H),1.47(m,9H). LCMS:[M+H-56] + =237.3.

[0305] To a solution of tert-butyl 3-(2-fluoro-1-hydroxyethyl)-3-nitropiperidine-1-carboxylate (287-2) (25 g, 85.53 mmol) in DCM (100 mL) was added BAST (85.15 g, 384.87 mmol) and stirred at 50° C. for 4 h. The mixture was diluted with NaHCO 3Quenched with aqueous solution (1.0 L) and extracted with EA (200 mL×3). The organic layer was concentrated in vacuo to give a residue. The residue was purified by column chromatography (PE:EA=100:1-20:1) to give tert-butyl 3-(1,2-difluoroethyl)-3-nitropiperidine-1-carboxylate (287-3). LCMS: [M+H-56] + =239.3.

[0306] To a solution of tert-butyl 3-(1,2-difluoroethyl)-3-nitropiperidine-1-carboxylate (287-3) (10.0 g, 33.98 mmol) in MeOH (100 mL) was added Pd / C (5.0 g, 10% purity) and heated with H 2 (15 Psi) at 40° C. for 16 h. The mixture was filtered and concentrated in vacuo to give crude tert-butyl 3-amino-3-(1,2-difluoroethyl)piperidine-1-carboxylate (287-4). LCMS: [M+H-56] + =209.3.

[0307] A solution of tert-butyl 3-amino-3-(1,2-difluoroethyl)piperidine-1-carboxylate (287-4) (8.1 g, 30.65 mmol) and benzaldehyde (4.88 g, 45.97 mmol) in EtOH (100 mL) was stirred at 40 °C for 1 h. 4 (5.8 g, 153.23 mmol) was added to the mixture and stirred at 40° C. for 16 h. The reaction was detected by LCMS. The mixture was diluted with NH 4 Quench with aqueous Cl (100 mL) and H 2 It was diluted with O (200 mL) and extracted with EA (50 mL x 3). The organic layer was concentrated in vacuo to give tert-butyl 3-(benzylamino)-3-(1,2-difluoroethyl)piperidine-1-carboxylate (287-5). LCMS: [M+H] + =355.4.

[0308] To a solution of tert-butyl 3-(benzylamino)-3-(1,2-difluoroethyl)piperidine-1-carboxylate (287-5) (7.0 g, 19.75 mmol) in DCM (20 mL) was added TFA (10 mL) and stirred at 40° C. for 3 h. The mixture was diluted with NaHCO 3 Quenched with aqueous solution (200 mL) and extracted with DCM (50 mL x 3). The organic layer was concentrated in vacuo to give 2,2,2-trifluoroacetaldehyde compound and N-benzyl-3-(1,2-difluoroethyl)piperidin-3-amine (1:1) (Intermediate 287-6). LCMS: [M+H] + =254.9.

[0309] Intermediate 288-9: 3-(6-methyl-1,4-dioxan-2-yl)piperidin-3-amine [ka] To a stirred solution of NaH (7.7 g, 180 mmol, 1.0 equiv) in anhydrous THF (120 mL) was added prop-2-en-1-ol (10.4 g, 180 mmol, 1.0 equiv) dropwise at 0° C. The reaction mixture was stirred for 15 min at 0° C. and for another 15 min at 15° C. Further, 2-chloroacetic acid was added in anhydrous to another suspension of NaH (7.7 g, 180 mmol, 1.0 equiv) and THF (60 mL) at 0° C. After stirring for 15 min at 0° C., the previously prepared solution was added and the reaction mixture was stirred for 12 h at 15° C. The reaction mixture was quenched with water (100 mL) and the aqueous layer was acidified and extracted with EA (200 mL×3) and anhydrous Na 2 SO 4 The mixture was dried at 40° C., filtered and concentrated to give 2-(allyloxy)acetic acid (288-1). 1 H NMR (400MHz, CDCl3) δ ppm 5.98-5.85 (m, 1H), 5.39-5.22 (m, 2H), 4.15 (s, 2H), 4.13 (m, 2H). LC-MS:[M+Na]+=161.1.

[0310] To a solution of 2-(allyloxy)acetic acid (288-1) (9.2 g, 79.23 mmol, 1.0 equiv) in anhydrous DCM (60 mL) was added DMF (115 mg, 1.58 mmol, 0.02 equiv) and (COCl)2 (12.1 g, 95.07 mmol, 1.2 equiv) dropwise at 15° C. The reaction mixture was stirred at 15° C. for 2 h. The reaction mixture was concentrated in vacuo to give crude 2-(allyloxy)acetyl chloride (288-2). LC-MS: [M+C 7 H 9 N-HCl+H]+=206.2.

[0311] To a solution of 1-benzyl 3-methylpiperidine-1,3-dicarboxylate (16.9 g, 60.94 mmol, 1.0 equiv) in anhydrous THF (170 mL) was added a solution of LiHMDS (61 mL, 60.94 mmol, 1.0 equiv) dropwise at -78 °C. The reaction mixture was stirred at -78 °C for 1 h, 2-(allyloxy)acetyl chloride (288-2) (8.2 g, 60.94 mmol, 1.0 equiv) was added dropwise at -78 °C, and the reaction was stirred at -78 °C for 1 h and at 15 °C for an additional 1 h. The reaction mixture was diluted with saturated NH 4 Quench with Cl (200 mL), extract with EA (100 mL × 3), and 2 SO 4 Drying at 40° C., filtration and concentration gave crude 1-benzyl 3-methyl 3-(2-(allyloxy)acetyl)piperidine-1,3-dicarboxylate (288-3). LC-MS: [M+H] + =376.2.

[0312] To a solution of 1-benzyl 3-(2-(allyloxy)acetyl)piperidine-1,3-dicarboxylate (288-3) (20.4 g, 53.34 mmol, 1.0 equiv) in MeOH (100 mL) was added NaBH 4 (2.0 g, 53.34 mmol, 1.0 equiv) was added portionwise at 15° C. The reaction was stirred at 15° C. for 3 h. The reaction mixture was diluted with saturated NH 4The mixture was quenched with Cl (100 mL), acidified with 2N HCl to PH=4-5, extracted with EtOAc (100 ml×3), and the combined organic phase was washed with water (100 ml), brine (100 mL), dried over anhydrous sodium sulfate, and concentrated in vacuo to give a residue. The residue was purified by column chromatography (SiO 2 , PE / EA=20:1 to 1:1) to give 1-benzyl 3-methyl 3-(2-(allyloxy)-1-hydroxyethyl)piperidine-1,3-dicarboxylate (288-4). LC-MS: [M+H] + =378.3.

[0313] To a solution of 1-benzyl 3-(2-(allyloxy)-1-hydroxyethyl)piperidine-1,3-dicarboxylate 3-(288-4) (10.4 g, 27.55 mmol, 1.0 equiv.) in dry MeCN (100 mL) was added NIS (10.6 g, 4.684 mmol, 1.7 equiv.). The reaction mixture was stirred at 90° C. for 3 h. The reaction mixture was diluted with saturated Na 2 SO 3 (50 mL), extracted with EA (100 mL × 3), and 2 SO 4 The residue was purified by column chromatography (SiO 2 , PE / EA=20 / 1 to 2 / 1) to give 1-benzyl 3-methyl 3-(6-(iodomethyl)-1,4-dioxan-2-yl)piperidine-1,3-dicarboxylate (288-5). LC-MS: [M+H] + =504.2.

[0314] A solution of 1-benzyl 3-methyl 3-(6-(iodomethyl)-1,4-dioxan-2-yl)piperidine-1,3-dicarboxylate (288-5) (4.3 g, 8.54 mmol, 1.0 equiv.) and n-Bu in anhydrous toluene (200 mL). 3 To a solution of SnH (2.5 g, 8.54 mmol, 1.0 equiv.), AIBN (140 mg, 0.85 mmol, 0.1 equiv.) was added and the reaction mixture was cooled to 5° C. 2 The reaction mixture was stirred under H 2The mixture was quenched with a solution of KF (0.5 g, 8.54 mmol, 1.0 equiv) in 2O (50 mL), diluted with water (100 mL), extracted with EA (100 mL × 3), and purified with anhydrous Na 2 SO 4 The residue was purified by column chromatography (SiO 2 , PE / EA=20 / 1 to 1 / 1) to give 1-benzyl 3-methyl 3-(6-methyl-1,4-dioxan-2-yl)piperidine-1,3-dicarboxylate (288-6). LC-MS: [M+H] + =378.3.

[0315] To a solution of 1-benzyl 3-methyl 3-(6-methyl-1,4-dioxan-2-yl)piperidine-1,3-dicarboxylate (288-6) (3.6 g, 9.54 mmol, 1.0 equiv) in MeOH (50 mL) was added H 2 LiOH H in O (50 mL) 2 A solution of MeOH (4.0 g, 95.38 mmol, 10.0 equiv) was added at room temperature. The reaction was stirred at room temperature for 16 h. The reaction mixture was concentrated in vacuo to remove most of the MeOH, diluted with 20 mL of water, and the aqueous phase was washed with EA (10 mL x 3). The aqueous phase was then acidified with 2 N HCl to PH = 3-4, extracted with EA (50 mL x 3), and washed with anhydrous Na 2 SO 4 Drying at 40° C. and concentration in vacuo afforded 1-((benzyloxy)carbonyl)-3-(6-methyl-1,4-dioxan-2-yl)piperidine-3-carboxylate (288-7). LC-MS: [M+H] + =364.3.

[0316] To a solution of 1-((benzyloxy)carbonyl)-3-(6-methyl-1,4-dioxan-2-yl)piperidine-3-carboxylate (288-7) (2.1 g, 5.78 mmol, 1.0 equiv.) in anhydrous THF (800 mL) was added TEA (1.7 g, 17.34 mmol, 3.0 equiv.) and DPPA (2.2 g, 8.09 mmol, 1.4 equiv.) at 20° C., the reaction mixture was stirred at 70° C. for 2 h, cooled to 20° C., and then purified by H2 A solution of KOH (1.0 g, 17.34 mmol, 3.0 equiv) in 2O (10 mL) was added. The reaction mixture was stirred at 20° C. for 2 h. The reaction mixture was concentrated, extracted with EA (50 mL×3), washed with water (50 mL), brine (50 mL). The organic layer was extracted with Na 2 SO 4 The crude product was purified by column chromatography (SiO 2 , PE / EA=20 / 1 to EA) to give impure product, which was further purified by preparative HPLC to give 3-amino-3-(6-methyl-1,4-dioxan-2-yl)piperidine-1-carboxylate benzyl (288-8). 1 H NMR (400 MHz, CDCl 3 )δ ppm 7.39-7.21(m,5H),5.06(m,2H),3.96-2.86(m,10H),1.82-1.61(m,2H),1.52-1.31(m,2H),1.30-0.96(m,3H). LC-MS:[M+H] + =335.3.

[0317] To a solution of benzyl 3-amino-3-(6-methyl-1,4-dioxan-2-yl)piperidine-1-carboxylate (288-8) (1.0 g, 2.99 mmol, 1.0 equiv) in MeOH (10 mL) was added Pd(OH) 2 (104 mg, 10 wt%) was added and the reaction mixture was stirred at 4°C for 2 h. 2 The mixture was stirred under atmosphere (16 Psi) for 8 hours at 20° C. The reaction mixture was filtered and concentrated in vacuo to give crude 3-(6-methyl-1,4-dioxan-2-yl)piperidin-3-amine (Intermediate 288-9).

[0318] Intermediate 292-3: 3-Oxa-1,7-diazaspiro[4.5]decan-2-one [ka] THF / MeOH / H 2To a solution of 1-benzyl 3-ethyl 3-(hydroxymethyl)piperidine-1,3-dicarboxylate (Intermediate 22-2) (2.0 g, 6.97 mmol, 1.0 equiv) in 2H2O (1:1:1, 30 mL), LiOH . H 2 O (0.87 g, 20.9 mmol, 3.0 equiv) was added at 20° C. The mixture was stirred at 20° C. for 8 h. Water (50 mL) was added and the mixture was extracted with EtOAc (100 mL×3). The aqueous phase was acidified to pH=2-3 with 1 M HCl and then extracted with EtOAc (100 mL×3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated to give 1-(tert-butoxycarbonyl)-3-(hydroxymethyl)piperidine-3-carboxylate (292-1). 1 H NMR (CDCl 3 400MHz):δ 4.05-3.85(m,1H),3.85-3.65(m,3H),3.45-3.20(m,1H),3.14-2.95(m,1 H),1.97-1.85(m,1H),1.80-1.70(m,1H),1.62-1.52(m,2H),1.44(s,9H).

[0319] To a solution of 1-(tert-butoxycarbonyl)-3-(hydroxymethyl)piperidine-3-carboxylate (292-1) (1.8 g, 6.94 mmol, 1.0 equiv.) in toluene (40 mL), TEA (1.4 g, 13.9 mmol, 2.0 equiv.) and DPPA (2.7 g, 10.4 mmol, 1.5 equiv.) were added in N 2 The reaction mixture was then stirred at 110° C. for 8 hours. 2 O (100 mL) was added and the mixture was extracted with EtOAc (100 mL×3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography (PE:EA=1:1) to give tert-butyl 2-oxo-3-oxa-1,7-diazaspiro[4.5]decane-7-carboxylate (292-2). 1 H NMR (CDCl 3400MHz):δ 5.87(brs,1H),4.22(d,J=8.8Hz,1H),4.07(d,J=8.8Hz,1H),3.66-3.42(m,2H),3.38 -3.18(m,2H),1.85-1.78(m,2H),1.75-1.65(m,1H),1.60-1.52(m,1H),1.48(s,9H).

[0320] A solution of tert-butyl 2-oxo-3-oxa-1,7-diazaspiro[4.5]decane-7-carboxylate (292-2) (0.6 g, 2.34 mmol, 1.0 equiv) in HCl / dioxane (5 mL, 4N) was stirred for 2 h at 20° C. The mixture was concentrated to give crude 3-oxa-1,7-diazaspiro[4.5]decan-2-one (intermediate 292-3).

[0321] Intermediate 293-8: 3-(1,1-difluoroprop-1-en-2-yl)piperidin-3-amine [ka] N 2 To a solution of tert-butyl 3-cyanopiperidine-1-carboxylate (25 g, 119.0 mmol, 1.0 equiv) in THF (300 mL) at −70° C. under atmospheric pressure was added LiHMDS (178 mL, 178.57 mmol, 1.5 equiv) dropwise. The reaction mixture was stirred at −70° C. for 30 min and then diluted with CH 3 CHO (7.85 g, 178.57 mmol, 3 equiv.) was added. The reaction was cooled to NH 4 It was quenched with aqueous Cl (200 mL) and extracted with EtOAc (100 mL×3). The combined organic layers were washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by Combi-flash (EtOAc in PE, 10%-50%) to give tert-butyl 3-(1-hydroxyethyl)-3-methylpiperidine-1-carboxylate (293-1).

[0322] To a solution of tert-butyl 3-(1-hydroxyethyl)-3-methylpiperidine-1-carboxylate (Intermediate 293-1) (9 g, 35.43 mmol, 1.0 equiv.) in DCM (150 mL) was added DMP (22.5 g, 53.15 mmol, 1.5 equiv.). The reaction mixture was stirred at 20° C. for 1 h. The reaction was cooled to 30° C. with NaHCO 3 (150mL) aqueous solution and Na 2 SO 3 (150 mL) and extracted with EtOAc (150 mL x 3). The combined organic layers were washed with NaHCO 3 Washing with aqueous solution (150 mL), drying over anhydrous sodium sulfate, filtering and concentrating gave tert-butyl 3-acetyl-3-cyanopiperidine-1-carboxylate (293-2). 1 H NMR (400 MHz, CDCl 3 )δ ppm 4.50-3.85(m,2H),3.20-3.05(m,1H),2.95-2.60(m,1H),2.46(s,3H),2.25-1.70(m,4H),1.48(s,9H).

[0323] To a solution of tert-butyl 3-acetyl-3-cyanopiperidine-1-carboxylate (293-2) (5 g, 19.84 mmol, 1.0 equiv) in DCM (100 mL) was added TFA (15 mL). The mixture was stirred at 20° C. for 2 h. The reaction mixture was then concentrated. The residue was diluted with EtOAc (150 mL) and diluted with NaHCO 3 Aqueous solution (150 mL) was added. Then, CbzCl (4.0 g, 23.81 mmol, 1.2 equiv.) was added. The reaction mixture was stirred at 20° C. for another 1 h. The product was extracted with EtOAc (50 mL×3). The combined organic layers were washed with brine (50 mL) and washed with anhydrous Na 2 SO 4 The residue was purified by combi-flash (EtOAc in PE, 0% to 30%) to give benzyl 3-acetyl-3-cyanopiperidine-1-carboxylate (293-3). 1 H NMR (400 MHz, CDCl 3)δ ppm 7.45-7.30(m,5H),5.25-5.10(m,2H),4.50-4.05(m,2H),3.20(d,J=13.6 MHz, 1H), 2.95-2.75 (m, 1H), 2.47 (d, J=16.8MHz, 3H), 2.25-1.60 (m, 4H).

[0324] To a solution of benzyl 3-acetyl-3-cyanopiperidine-1-carboxylate (293-3) (5 g, 17.46 mmol, 1.0 equiv.) and 2-((difluoromethyl)sulfonyl)pyridine (5 g, 26.19 mmol, 1.5 equiv.) in THF (50 mL) was added KHMDS (26 mL, 26.19 mmol, 1.5 equiv.) and N 2 The reaction mixture was stirred at -70°C for 2 hours. The reaction mixture was diluted with NH 4 The mixture was quenched with aqueous Cl (50 mL) and extracted with EtOAc (50 mL x 3). The organic layer was washed with brine (100 mL) and anhydrous Na 2 SO 4 Drying at 40° C., filtration and concentration gave 3-cyano-3-(1,1-difluoro-2-hydroxy-1-(pyridin-2-ylsulfonyl)propan-2-yl)piperidine-1-carboxylate (293-4). LC-MS: [M+H] + =480.0.

[0325] To a solution of benzyl 3-cyano-3-(1,1-difluoro-2-hydroxy-1-(pyridin-2-ylsulfonyl)propan-2-yl)piperidine-1-carboxylate (293-4) (12 g, crude) in DMF (30 mL) was added saturated NH 4 Cl (30 mL) and aqueous HCl (30 mL, 6 M) were added. The reaction mixture was stirred at 80° C. for 48 h. The reaction was cooled and 2 The mixture was diluted with 200 mL of EtOAc (50 mL×3). The mixture was extracted with EtOAc (50 mL×3). The combined organic layers were washed with brine (100 mL) and diluted with Na 2 SO 4The residue was purified by combi-flash (EtOAc in PE, 0% to 20%) to give benzyl 3-cyano-3-(1,1-difluoroprop-1-en-2-yl)piperidine-1-carboxylate (293-5). 1 H NMR (400 MHz, CDCl 3 )δ ppm 7.45-7.30(m,5H),5.30-5.15(m,2H),4.65-4.15(m,2H),3.10-2.70(m,2H),2.25(d,J=11.2MHz,1H),2.10-1.60(m,6H). LC-MS:[M+H] + =320.9.

[0326] To a mixture of benzyl 3-cyano-3-(1,1-difluoroprop-1-en-2-yl)piperidine-1-carboxylate (293-5) (2.7 g, 8.44 mmol, 1.0 equiv.) in MeOH (100 mL), 1 M NaOH solution (12.6 mL, 1.5 equiv.) was added H 2 O 2 (30%, 6.68 g, 59 mmol, 7.0 equiv) was added. The reaction was stirred at 18° C. for 36 h. The reaction was diluted with saturated Na 2 S 2 O 3 (50 mL). The aqueous layer was extracted with EtOAc (50 mL×3). The combined organic layers were washed with Na 2 SO 4 The residue was purified by combi-flash (EtOAc in PE, 0-40%) to give benzyl 3-carbamoyl-3-(1,1-difluoroprop-1-en-2-yl)piperidine-1-carboxylate (293-6). 1 H NMR (CDCl 3 400MHz): δ ppm 7.42-7.30(m,5H),5.22-5.08(m,2H),4.48(brs,1H),3.99(brs,1H),3.17-2.88(m,2H),2.53(brs,1H),1.76-1.46(m,7H).

[0327] To a solution of benzyl 3-carbamoyl-3-(1,1-difluoroprop-1-en-2-yl)piperidine-1-carboxylate (293-6) (1.34 g, 4 mmol, 1.0 equiv.) and KOH (560 mg, 10 mmol, 2.5 equiv.) in MeOH (30 mL), PhI(OAc) 2 (1.53 g, 4.8 mmol, 1.2 equiv) was added at 0° C. The resulting mixture was stirred at 15° C. for 3 h. The reaction was diluted with saturated NH 4 The mixture was quenched with aqueous Cl (50 mL). The aqueous layer was extracted with EtOAc (50 mL×3). The combined organic layers were washed with Na 2 SO 4 The residue was purified by combi-flash (EtOAc in PE, 0-30%) to give benzyl 3-(1,1-difluoroprop-1-en-2-yl)-3-((methoxycarbonyl)amino)piperidine-1-carboxylate (293-7). 1 H NMR (400 MHz, CDCl 3 )δ ppm 7.42-7.30(m,5H),5.24-5.08(m,2H),4.30(d,J=14.2Hz,1H),4.18-4.04(m,1H),3.60(s,3H),2.82(brs,2H),1.73-1.60(m,4H),1.57(s,3H). LC-MS:[M+H] + =369.0.

[0328] To a solution of benzyl 3-(1,1-difluoroprop-1-en-2-yl)-3-((methoxycarbonyl)amino)piperidine-1-carboxylate (293-7) (600 mg, 1.63 mmol, 1.0 equiv) in DCM (15 mL) was added TMSI (977 mg, 4.89 mmol, 3.0 equiv). The reaction was stirred at 16 °C for 18 h. The reaction was quenched with water (10 mL). The aqueous layer was adjusted to pH = 4-5 with 1 M HCl solution. The mixture was partitioned between DCM and water. The aqueous layer was lyophilized to give 3-(1,1-difluoroprop-1-en-2-yl)piperidin-3-amine HCl salt (intermediate 293-8). 1 H NMR (400MHz, DMSO-d 6)δ ppm 3.73(d,J=13.4Hz,1H),3.17(d,J=12.8Hz,2H),2.98(t,J=10.2Hz,1H),2.27( d,J=13.6Hz,1H),2.01-1.78(m,2H),1.71(t,J=3.4Hz,3H),1.65-1.50(m,1H). LC-MS:[M+H] + =177.1

[0329] Intermediate 295-6: 3-(1-fluoroethyl)piperidin-3-amine [ka] A solution of 1-benzyl 3-methyl 3-((tert-butoxycarbonyl)amino)piperidine-1,3-dicarboxylate (160 mg, 0.41 mmol) in MeOH (10 mL) was treated with LiCl (510 mg, 12.3 mmol) and NaBH 4 (462 mg, 12.3 mmol) was added at room temperature. The reaction mixture was stirred at this temperature until no starting material remained. The reaction mixture was diluted with EA and washed with 1N HCl and brine. The organic layer was extracted with Na 2 SO 4 and concentrated in vacuo to give the crude alcohol. LC-MS: [M+H-100] + =265.0.

[0330] To a solution of the alcohol (560 mg, 1.537 mmol) in DCM (15 mL) was added Dess-Martin reagent (782 mg, 1.844 mmol). The mixture was stirred at room temperature for 2 h. The mixture was diluted with NaHCO 3 aqueous solution (10 mL) and the layers were separated. The aqueous layer was extracted with DCM (10 ml x 2). The combined organic layers were washed with brine and 2 SO 4 The mixture was dried over 1000 ml and concentrated. The filtrate was concentrated. The residue was purified and eluted with EA (0-50%) in n-hexane to give benzyl 3-((tert-butoxycarbonyl)amino)-3-formylpiperidine-1-carboxylate (295-1). LC-MS: [M+H-100] + =624.8.

[0331] A solution of benzyl 3-((tert-butoxycarbonyl)amino)-3-formylpiperidine-1-carboxylate (295-1) (100 mg, 0.276 mmol) in THF (10 mL) was diluted with CH 3 Aqueous MgBr was added dropwise at -78°C. The mixture was stirred at -78°C for an additional 1 h and warmed to 0°C for an additional 2 h. The mixture was diluted with NH 4 The mixture was quenched with aqueous Cl (10 ml) and extracted with EA (20 ml×3). The combined organic layers were washed with brine and 2 SO 4 The residue was purified and eluted with EA (0-50%) in n-hexane to give the alcohol. LC-MS: [M+H-100] + = 279.0. To a solution of the alcohol (100 mg, 0.264 mmol) in DCM (3 mL) was added TFA (0.5 mL). The resulting mixture was stirred at room temperature for 2 h. The mixture was concentrated under vacuum (below 40°C) to give benzyl 3-amino-3-(1-hydroxyethyl)piperidine-1-carboxylate (295-2). LC-MS [M+H] + =279.0.

[0332] To a solution of benzyl 3-amino-3-(1-hydroxyethyl)piperidine-1-carboxylate (295-2) (330 mg, 1.186 mmol), DMAP (14.48 mg, 0.119 mmol), and DIPEA (0.958 mL, 5.34 mmol) in DCM (20 mL) was added NsCl (657 mg, 2.96 mmol) in portions. The mixture was cooled to 5° C. for 1 hour and then cooled to 5° C. 2 The reaction was stirred overnight at room temperature under NaHCO 3 aqueous solution (20 mL). The layers were separated. The aqueous layer was extracted with DCM (20 mL x 2). The combined organic layers were washed with brine and 2 SO 4The mixture was dried over 1000 ml and concentrated. The filtrate was concentrated. The residue was purified and eluted with n-hexane in EA (0-30%) to give benzyl 2-methyl-1-((4-nitrophenyl)sulfonyl)-1,5-diazaspiro[2.5]octane-5-carboxylate (295-3). LC-MS: [M+H] + =445.8.

[0333] To a solution of benzyl 2-methyl-1-((4-nitrophenyl)sulfonyl)-1,5-diazaspiro[2.5]octane-5-carboxylate (295-3) (420 mg, 0.943 mmol) in THF (10 mL), 1 M TBAF in THF (0.606 ml, 0.606 mmol) was added 2 The mixture was heated to 45° C. overnight. The mixture was cooled to room temperature and added EA (15 mL) and H 2 The layers were separated and the aqueous layer was extracted with EA (15 mL×3). The combined organic layers were washed with brine and then with Na 2 SO 4 The mixture was dried over 1000 ml of ethyl acetate and filtered. The filtrate was concentrated and purified by preparative HPLC to give benzyl 3-(1-fluoroethyl)-3-((4-nitrophenyl)sulfonamido)piperidine-1-carboxylate (295-4). LC-MS: [M+H] + =465.8.

[0334] CH 3 A mixture of thioglycolic acid (71.2 mg, 0.773 mmol) and DBU235 (98 mg, 1.547 mmol) in CN (10 mL) was 2 The mixture was stirred under reduced pressure for 5 min. Then, 3 Benzyl 3-(1-fluoroethyl)-3-((4-nitrophenyl)sulfonamido)piperidine-1-carboxylate (295-4) (120 mg, 0.258 mmol) in CN (2 mL) was added. The mixture was stirred at room temperature overnight and concentrated. The residue was treated with EA (10 mL) and NaHCO 3 Aqueous solution (5mL x 2), H 2 20 mL, washed with saline, and then with Na 2 SO 4The mixture was dried over 1000 ml and filtered. The filtrate was concentrated and the residue was purified by preparative HPLC to give benzyl 3-amino-3-(1-fluoroethyl)piperidine-1-carboxylate (295-5). LC-MS: [M+H] + =281.1.

[0335] To a solution of benzyl 3-amino-3-(1-fluoroethyl)piperidine-1-carboxylate (295-5) (120 mg, 0.428 mmol) in MeOH (15 ml), Pd(OH)2 (60 mg) was added with N 2 The mixture was then added under H 2 Stirred under balloon for 3 hours. The mixture was filtered and the filtrate was concentrated to give 3-(1-fluoroethyl)piperidin-3-amine (Intermediate 295-6). LC-MS: [M+H] + =147.1

[0336] Intermediate 297-6: 3-((methylthio)methyl)piperidin-3-amine [ka] To a solution of 1-benzyl 3-ethyl 3-((((trifluoromethyl)sulfonyl)oxy)methyl)piperidine-1,3-dicarboxylate (22-3) (18.0 g, 39.7 mmol, 1.0 equiv) in THF (200 mL) was added MeSNa (11.0 g, 159 mmol, 4.0 equiv) at 0 °C. The resulting mixture was stirred at 15-20 °C for 16 h. The reaction was quenched with water (300 mL) and extracted with EtOAc (300 mL × 3). The combined organic layer was washed with anhydrous Na 2 SO 4 The residue was purified by Combi Flash (30% EtOAc in PE) to give 1-benzyl 3-ethyl 3-((methylthio)methyl)piperidine-1,3-dicarboxylate (297-1). 1 H NMR (400 MHz, CDCl 3)δ ppm 7.25-7.45(5H,m),5.13(2H,s),4.05-4.25(2H,m),3.80-4.00(1H,m),3.25-3.60( 3H, m), 2.60-2.85 (2H, m), 2.00-2.15 (4H, m), 1.55-1.75 (3H, m), 1.10-1.30 (3H, m).

[0337] MeOH / THF / H 2 1-Benzyl 3-ethyl 3-((methylthio)methyl)piperidine-1,3-dicarboxylate (297-1) (10.0 g, 28.5 mmol, 1.0 equiv.) in 2O (120 mL, v / v / v=1 / 1 / 1) and LiOH . H 2 A mixture of 2H2O (12.0 g, 285 mmol, 10.0 equiv.) was stirred at 20° C. for 16 h. The reaction was concentrated in vacuo. The residue was diluted with water (200 mL) and extracted with MTBE (100 mL). The pH of the aqueous layer was then adjusted to 5 with 1H HCl and extracted with EtOAc (200 mL×3). The combined organic layers were washed with anhydrous Na 2 SO 4 and concentrated in vacuo to give crude 1-((benzyloxy)carbonyl)-3-((methylthio)methyl)piperidine-3-carboxylate (297-2). LC-MS: [M+H] + =324.1.

[0338] 1-((benzyloxy)carbonyl)-3-((methylthio)methyl)piperidine-3-carboxylate (297-2) (8.00 g, 24.7 mmol, 1.0 equiv.), DPPA (7.50 g, 27.2 mmol, 1.1 equiv.), and Et 3 N (5.00 g, 48.5 mmol, 2.0 equiv.) 2 The mixture was stirred at 80° C. for 1 h under reduced pressure. The reaction was then concentrated in vacuo. The residue was dissolved in dioxane (50 mL) and 6N HCl (50 mL) and the mixture was stirred at 25-30° C. for 16 h. The reaction was then concentrated in vacuo. The pH of the aqueous layer was adjusted to 9 with 1N NaOH. The mixture was then extracted with EtOAc (200 mL×3). The combined organic layers were washed with anhydrous Na 2SO 4 The residue was dissolved in DCM (100 mL) and diluted with DIPEA (10.0 g, 74.2 mmol, 3.0 equiv.) and Boc 2 0 (11.0 g, 48.4 mmol) was added. The resulting mixture was stirred at 25-30 °C for 16 h. The reaction was concentrated in vacuo. The residue was purified by Combi Flash (20% EtOAc in PE) to give benzyl 3-((tert-butoxycarbonyl)amino)-3-((methylthio)methyl)piperidine-1-carboxylate (297-5). LC-MS: [M+H-100] + =295.1.

[0339] CH 3 To a solution of benzyl 3-((tert-butoxycarbonyl)amino)-3-((methylthio)methyl)piperidine-1-carboxylate (297-5) (2.37 g, 6.00 mmol, 1 equiv.) in CN (50 mL) was added TMSI (3.60 g, 18 mmol, 3 equiv.) dropwise at about 0° C. After addition, the reaction mixture was stirred at the same temperature for 2 h. The reaction mixture was carefully quenched with MeOH (20 mL) and HCl / dioxane (5 mL, 4 M). The reaction mixture was concentrated. Water (20 mL) was added and the mixture was extracted with MTBE (40 mL×4). The aqueous layer was lyophilized to give crude 3-((methylthio)methyl)piperidin-3-amine (intermediate 297-6).

[0340] Intermediate 299-3: 2,2,2-trifluoroacetaldehyde compound and 1-(3-aminopiperidin-3-yl)-2,2,2-trifluoroethan-1-ol (1:1) [ka] To a mixture of tert-butyl 3-nitropiperidine-1-carboxylate (5.0 g, 21.71 mmol, 1.0 equiv.) and 2,2,2-trifluoroethane-1,1-diol (10.08 g, 86.86 mmol, 4.0 equiv.) in THF (50 mL) was added K 2 CO 3(1.8 g, 13.03 mmol, 0.6 equiv) was added at 60° C. for 16 h. 1N HCl (10 mL) was added to the mixture and extracted with EA (40 mL×3). The combined organic layers were washed with brine (30 mL) and diluted with Na 2 SO 4 Dry at 40° C., filter and concentrate in vacuo to give tert-butyl 3-nitro-3-(2,2,2-trifluoro-1-hydroxyethyl)piperidine-1-carboxylate (299-1). H NMR: 1 H NMR (400 MHz, CDCl 3 )δ ppm 5.01-4.71(m,2H),4.67-4.34(m,1H),4.07-3.35(m,2H),2.88-2.13(m,2H),1.95-1.52(m,2H),1.50-1.36(m,9H). LCMS:[M+H-56] + =273.3.

[0341] EtOH / H 2 tert-Butyl 3-nitro-3-(2,2,2-trifluoro-1-hydroxyethyl)piperidine-1-carboxylate (299-1) (4.0 g, 12.89 mmol, 1.0 equiv.) and NH 4 To a solution of Cl (3.45 g, 64.45 mmol, 5.0 equiv.), Zn (8.43 g, 128.91 mmol, 10.0 equiv.) was added 2 The mixture was stirred at 80° C. for 16 h, worked up under aqueous conditions, filtered and concentrated in vacuo. The crude was purified by column chromatography (PE:EA=10:1) to give tert-butyl 3-amino-3-(2,2,2-trifluoro-1-hydroxyethyl)piperidine-1-carboxylate (299-2). H NMR: 1 H NMR (400 MHz, CDCl 3 )δ ppm 4.05-3.93(m,1H),3.69-3.50(m,2H),3.22-2.74(m,2H),1.90-1.70(m,2H),1.68-1.60(m,2H),1.47(s,9H).

[0342] To a solution of tert-butyl 3-amino-3-(2,2,2-trifluoro-1-hydroxyethyl)piperidine-1-carboxylate (299-2) (1.2 g, crude) in DCM (10 mL), TFA (2 mL) was added dropwise under nitrogen protection, and the mixture was stirred for 2 h at 25° C. The mixture was concentrated to give 2,2,2-trifluoroacetaldehyde compound and 1-(3-aminopiperidin-3-yl)-2,2,2-trifluoroethan-1-ol (1:1) (intermediate 299-3).

[0343] Intermediate 300-6: (5-(3-amino-3-(1,2,2-trifluoroethyl)piperidin-1-yl)-2-(3,4-difluorophenyl)pyridin-4-yl)methanol [ka] To a solution of tert-butyl 3-nitropiperidine-1-carboxylate (3 g, 13.03 mmol) in THF (30 mL), 2 CO 3 (0.540 g, 3.91 mmol) and 1-ethoxy-2,2-difluoroethanol (4.93 g, 39.1 mmol) were added and the reaction mixture was heated at 55 °C with N 2 The mixture was stirred under atmospheric pressure for 18 hours. The reaction mixture was diluted with water and extracted with EtOAc (20 mL×3), and the combined organic phase was washed with water (20 ml), brine (20 ml), dried over anhydrous sodium sulfate, and concentrated in vacuo to give a residue. The residue was purified by flash chromatography (elution gradient: 0% to 40% EtOAc in PE, 30 min) to give tert-butyl 3-(2,2-difluoro-1-hydroxyethyl)-3-nitropiperidine-1-carboxylate (300-1). LC-MS: [M+H] + =311.1

[0344] A solution of tert-butyl 3-(2,2-difluoro-1-hydroxyethyl)-3-nitropiperidine-1-carboxylate (300-1) (2.0 g, 6.45 mmol) in BAST (20 mL) was diluted with N 2The mixture was stirred under atmospheric pressure at 60° C. for 4 hours. The reaction mixture was diluted with water, extracted with EtOAc, and the combined organic phase was washed with water, saturated NaHCO 3 The solution was washed with brine, dried over sodium sulfate and concentrated to give the crude product. The reaction mixture was diluted with water and extracted with EtOAc (20ml x 3), and the combined organic phase was washed with water (20ml), brine (20ml), dried over anhydrous sodium sulfate and concentrated in vacuo to give a residue. The crude product was purified by flash chromatography (elution gradient: 0% to 30% EtOAc in PE, 30 min) to give tert-butyl 3-nitro-3-(1,2,2-trifluoroethyl)piperidine-1-carboxylate (300-2). LC-MS: [M+H] + =313.1

[0345] To a solution of tert-butyl 3-nitro-3-(1,2,2-trifluoroethyl)piperidine-1-carboxylate (300-2) (1.36 g, 4.35 mmol) in EtOH (30 mL) and water (15 mL) was added Fe (2.432 g, 43.5 mmol) and NH 4 Cl (1.165 g, 21.77 mmol) was added and the reaction mixture was cooled to 10° C. 2 The mixture was stirred at 60° C. under ambient atmosphere for 18 hours. The reaction mixture was filtered, the filtrate was diluted with water, extracted with EtOAc (30 ml×3), and the combined organic phase was washed with water (20 ml), brine (20 ml), dried over anhydrous sodium sulfate, and concentrated in vacuo to give a residue. The crude product was purified by flash chromatography (elution gradient: 0% to 50% EtOAc in PE, 30 min) to give tert-butyl 3-amino-3-(1,2,2-trifluoroethyl)piperidine-1-carboxylate (300-3). LC-MS: [M+H] + =283.2.

[0346] To a solution of tert-butyl 3-amino-3-(1,2,2-trifluoroethyl)piperidine-1-carboxylate (300-3) (700 mg, 2.480 mmol) in DCM (12 mL) was added TFA (4 mL) and the reaction mixture was purified by N 2The mixture was stirred at room temperature under atmospheric pressure for 1 hour. The reaction mixture was concentrated to give 3-(1,2,2-trifluoroethyl)piperidin-3-amine (300-4). LC-MS: [M+H] + =183.2.

[0347] To a solution of methyl 2-(3,4-difluorophenyl)-5-fluoroisonicotinate (Intermediate A) (0.847 g, 3.17 mmol) and 3-(1,2,2-trifluoroethyl)piperidin-3-amine (300-4) (1.3 g, 3.17 mmol) in DMSO (30 mL) was added DIPEA (15 mL) at room temperature and the reaction mixture was cooled to 5° C. 2 The mixture was stirred at 80° C. under ambient atmosphere for 8 hours. The reaction mixture was diluted with water, extracted with EtOAc (20 ml×3), worked up under aqueous conditions, and concentrated in vacuo to give a residue. The crude product was purified by flash chromatography (elution gradient: 0% to 30% EtOAc in PE, 30 min) to give methyl 5-(3-amino-3-(1,2,2-trifluoroethyl)piperidin-1-yl)-2-(3,4-difluorophenyl)isonicotinate (300-5). LC-MS: [M+H] + =430.1.

[0348] To a solution of methyl (5-(3-amino-3-(1,2,2-trifluoroethyl)piperidin-1-yl)-2-(3,4-difluorophenyl)isonicotinate 300-5) (0.65 g, 1.514 mmol) in THF (20 mL) and methanol (20 mL) was added lithium chloride (1.284 g, 30.3 mmol) and sodium tetrahydroborate (1.145 g, 30.3 mmol) at room temperature, and the reaction mixture was diluted with N 2The mixture was stirred at room temperature under atmospheric pressure for 3 hours. The reaction mixture was diluted with water and extracted with EtOAc (20 mL×3), and the combined organic phase was washed with water (20 mL), brine (20 mL), dried over anhydrous sodium sulfate, and concentrated in vacuo to give a residue. The crude product was purified by flash chromatography (elution gradient: 10% to 40% EtOAc in PE, 30 min) to give (5-(3-amino-3-(1,2,2-trifluoroethyl)piperidin-1-yl)-2-(3,4-difluorophenyl)pyridin-4-yl)methanol (Intermediate 300-6). LC-MS: [M+H] + =402.2.

[0349] Intermediate 300-7 [ka] To a solution of (5-(3-amino-3-(1,2,2-trifluoroethyl)piperidin-1-yl)-2-(3,4-difluorophenyl)pyridin-4-yl)methanol (Intermediate 300-6) (500 mg, 1.246 mmol), Intermediate B (501 mg, 1.495 mmol), and triphenylphosphine (980 mg, 3.74 mmol) in THF (90 mL) was added DEAD (0.592 mL, 3.74 mmol) dropwise at 0 °C and the reaction mixture was quenched with N 2 The mixture was stirred at 0° C. under ambient atmosphere for 0.5 h. The reaction mixture was diluted with water and extracted with EtOAc (20 ml×3), and the combined organic phase was washed with water (20 ml), brine (20 ml), dried over anhydrous sodium sulfate, and concentrated in vacuo to give a residue. The crude product was purified by flash chromatography (elution gradient: 0%-10% MeOH in DCM, 50 min) to give intermediate 300-7. LC-MS: [M+H] + =719.2.

[0350] Intermediate 301-8: 3-amino-1-(4-(chloromethyl)-6-(3,4-difluorophenyl)pyridin-3-yl)-3-(2,2-difluoroethyl)piperidin-4-ol [ka] A solution of LDA (6.94 mL, 13.88 mmol) in THF (9 mL) was diluted with a solution of 1-(tert-butyl) 3-methyl 4-hydroxypiperidine-1,3-dicarboxylate (1.5 g, 5.78 mmol) in THF with N 2 After addition under atmospheric pressure at −78° C. and stirring at this temperature for 10 min, a solution of 1,1-difluoro-2-iodoethane (1.443 g, 7.52 mmol) in HMPA (5 mL, 28.7 mmol) was added at −15° C. and the reaction mixture was immersed in N 2 The mixture was stirred at room temperature under atmospheric pressure for 30 minutes. The reaction mixture was diluted with water and extracted with EtOAc (20 ml x 3), and the combined organic phase was washed with water (20 ml), brine (20 ml), dried over anhydrous sodium sulfate, and concentrated in vacuo to give a residue. The residue was purified by flash chromatography (elution gradient: 0% to 30% EtOAc in PE, 30 min) to give 1-(tert-butyl) 3-methyl 3-(2,2-difluoroethyl)-4-hydroxypiperidine-1,3-dicarboxylate (301-1). LC-MS: [M+H] + =324.1.

[0351] A solution of 1-(tert-butyl) 3-methyl 3-(2,2-difluoroethyl)-4-hydroxypiperidine-1,3-dicarboxylate (301-1) (0.6 g, 1.856 mmol) in methanol (5 mL) was diluted with a solution of LiOH (700 mg, 29.2 mmol) in water (5 mL) and diluted with N 2 The mixture was stirred at room temperature for 5 hours under atmospheric pressure. The pH of the reaction mixture was adjusted to 4 by adding 3M HCl solution, extracted with EtOAc (20 ml x 3), and the combined organic phase was washed with water (20 ml), brine (20 ml), dried over anhydrous sodium sulfate, and concentrated in vacuo to give crude 1-(tert-butoxycarbonyl)-3-(2,2-difluoroethyl)-4-hydroxypiperidine-3-carboxylate (301-2). LC-MS: [M+H] + =310.1.

[0352] To a solution of 1-(tert-butoxycarbonyl)-3-(2,2-difluoroethyl)-4-hydroxypiperidine-3-carboxylic acid (301-2) (0.4 g, 1.293 mmol) in toluene (10 mL), DPPA (0.534 g, 1.940 mmol) and TEA (0.721 mL, 5.17 mmol) were added with N 2 The mixture was added at room temperature under atmospheric pressure and the reaction was stirred at 100° C. for 18 hours. The reaction mixture was diluted with water and extracted with EtOAc (20 ml×3), and the combined organic phase was washed with water (20 ml), brine (20 ml), dried over anhydrous sodium sulfate, and concentrated in vacuo to give a residue. The residue was purified by flash chromatography (elution gradient: 0% to 50% EtOAc in PE, 30 min) to give tert-butyl 3a-(2,2-difluoroethyl)-2-oxohexahydrooxazolo[4,5-c]pyridine-5(4H)-carboxylate (303-3). 1 H NMR (400MHz, DMSO-d 6 )δ 8.12(s,1H),6.43-5.89(m,1H),4.41-4.15(m,2H),4.13(dt,J=12.9,2.5Hz,1H),3.03-2.75(m,2H),2.03-1.74(m,4H),1.39(s,9H). LC-MS:[M+H] + =307.1.

[0353] To a solution of tert-butyl 3a-(2,2-difluoroethyl)-2-oxohexahydrooxazolo[4,5-c]pyridine-5(4H)-carboxylate (301-3) (400 mg, 1.306 mmol) in DCM (9 mL) was added TFA (3 mL, 38.9 mmol) 2 The mixture was added at room temperature under atmospheric pressure and the reaction was stirred at room temperature for 1 hour. The reaction mixture was concentrated in vacuo to give crude 3a-(2,2-difluoroethyl)hexahydrooxazolo[4,5-c]pyridin-2(3H)-one (301-4). LC-MS: [M+H] + =207.1.

[0354] To a solution of methyl 2-(3,4-difluorophenyl)-5-fluoroisonicotinate (Intermediate A) (259 mg, 0.970 mmol) and 3a-(2,2-difluoroethyl)hexahydrooxazolo[4,5-c]pyridin-2(3H)-one (301-4) in DMSO (10 mL), DIPEA (10 mL, 57.3 mmol) was added with N 2 The mixture was added at room temperature under atmospheric pressure and the reaction was stirred at 120° C. for 18 hours. The reaction mixture was diluted with water and extracted with EtOAc (20 ml×3), and the combined organic phase was washed with water (20 ml×3), brine (20 ml), dried over anhydrous sodium sulfate, and concentrated in vacuo to give a residue. The residue was purified by flash chromatography (elution gradient: 0% to 10% MeOH in DCM, 30 min) to give methyl 5-(3a-(2,2-difluoroethyl)-2-oxohexahydrooxazolo[4,5-c]pyridin-5(4H)-yl)-2-(3,4-difluorophenyl)isonicotinate (301-5). LC-MS: [M+H] + =454.1.

[0355] To a solution of methyl 5-(3a-(2,2-difluoroethyl)-2-oxohexahydrooxazolo[4,5-c]pyridin-5(4H)-yl)-2-(3,4-difluorophenyl)isonicotinate (301-5) (40 mg, 0.088 mmol) in THF (5 mL) and methanol (5 mL) was added LiCl (74.8 mg, 1.764 mmol) and NaBH 4 (33.4 mg, 0.882 mmol) 2 The mixture was added at room temperature under atmospheric pressure and the reaction was stirred at room temperature for 5 hours. The reaction mixture was diluted with water and extracted with EtOAc (20 ml x 3), and the combined organic phase was washed with water (20 ml), brine (20 ml), dried over anhydrous sodium sulfate, and concentrated in vacuo to give a residue. The residue was purified by flash chromatography (elution gradient: 0% to 10% MeOH in DCM, 30 min) to give 3a-(2,2-difluoroethyl)-5-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridin-3-yl)hexahydrooxazolo[4,5-c]pyridin-2(3H)-one (301-6). LC-MS: [M+H]+ =426.2.

[0356] To a solution of 3a-(2,2-difluoroethyl)-5-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridin-3-yl)hexahydrooxazolo[4,5-c]pyridin-2(3H)-one (301-6) (100 mg, 0.235 mmol) in MeOH (10 mL) and water (10 mL) was added KOH (132 mg, 2.351 mmol) and N 2 At room temperature under atmospheric pressure, the reaction was stirred at 100° C. for 18 hours. The reaction mixture was diluted with water, extracted with EtOAc (20 ml×3), and the combined organic phase was washed with water (20 ml), brine (20 ml), dried over anhydrous sodium sulfate, and concentrated in vacuo to give a residue. The residue was purified by flash chromatography (0% to 10% MeOH in DCM, 30 min elution gradient) to give 3-amino-3-(2,2-difluoroethyl)-1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridin-3-yl)piperidin-4-ol (301-7). LC-MS: [M+H] + =400.1.

[0357] To a solution of 3-amino-3-(2,2-difluoroethyl)-1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridin-3-yl)piperidin-4-ol (301-7) (130 mg, 0.325 mmol) in DCM (10 mL) was added thionyl chloride (0.119 mL, 1.627 mmol) with N 2 The reaction was stirred at room temperature for 5 hours under atmospheric conditions at 0° C. The reaction was diluted with 5 mL of saturated K 2 CO 3 The reaction mixture was quenched with aqueous solution at 0° C. The reaction mixture was extracted with DCM (20 mL×3) and the combined organic phase was washed with water (20 mL), brine (20 mL), dried over anhydrous sodium sulfate and concentrated in vacuo to give crude 3-amino-1-(4-(chloromethyl)-6-(3,4-difluorophenyl)pyridin-3-yl)-3-(2,2-difluoroethyl)piperidin-4-ol (Intermediate 301-8). LC-MS: [M+H] + =418.1.

[0358] Intermediate 302-9: 3-amino-3-(6-chloropyridin-2-yl)-1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridin-3-yl)piperidin-4-ol [ka] To a solution of m-CPBA (23.17 g, 94 mmol) and 2-chloro-6-iodopyridine (15 g, 62.6 mmol) in DCM (450 mL) was added TfOH (22.25 mL, 251 mmol) dropwise at 0° C. The reaction mixture was then stirred at room temperature for 2 h, mesitylene (9.54 mL, 68.9 mmol) was added dropwise at 0° C., and the reaction mixture was purified by N 2 The mixture was stirred at room temperature under atmospheric pressure for 18 hours. The solvent was removed in vacuo to give the crude product. The crude product was dissolved in Et 2 Recrystallization in O at -20 °C gave (6-chloropyridin-2-yl)(mesityl)iodonium trifluoromethanesulfonate (302-1). LC-MS: [M+H] + =358.

[0359] To a solution of 1-tert-butyl 3-methyl 4-oxopiperidine-1,3-dicarboxylate (1.3 g, 5.05 mmol) in DMF (30 mL), potassium tert-butoxide (0.680 g, 6.06 mmol) was added at room temperature, and the reaction mixture was stirred at room temperature for 10 min, after which (6-chloropyridin-2-yl)(mesityl)iodonium trifluoromethanesulfonate (Intermediate 302-1) (3.52 g, 5.56 mmol) was added at room temperature, and the reaction mixture was cooled to 5° C. 2 The mixture was stirred at room temperature under atmosphere for 18 hours. The reaction mixture was diluted with water and extracted with EtOAc, and the combined organic phase was washed with water, brine, dried over sodium sulfate, and concentrated to give the crude product. The crude product was purified by flash chromatography (elution gradient: 0% to 30% EtOAC in hexanes, 30 min) to give 1-(tert-butyl) 3-methyl 3-(6-chloropyridin-2-yl)-4-oxopiperidine-1,3-dicarboxylate (302-2). LC-MS: [M+H]+ =369.0

[0360] To a solution of 1-(tert-butyl) 3-methyl 3-(6-chloropyridin-2-yl)-4-oxopiperidine-1,3-dicarboxylate (intermediate 302-2) (1.0 g, 2.71 mmol) in MeOH (30 mL) was added sodium tetrahydroborate (0.205 g, 5.42 mmol) at 0° C., and the reaction mixture was stirred at 0° C. for 30 min. The reaction mixture was diluted with water, extracted with EtOAc, and the combined organic phase was washed with water, brine, dried over sodium sulfate, and concentrated to give the crude product. The crude product was purified by flash chromatography (elution gradient: 0% to 30% EtOAC in hexanes, 30 min) to give 1-(tert-butyl) 3-methyl 3-(6-chloropyridin-2-yl)-4-hydroxypiperidine-1,3-dicarboxylate (302-3). LC-MS: [M+H] + =371.0

[0361] To a solution of 1-(tert-butyl) 3-methyl 3-(6-chloropyridin-2-yl)-4-hydroxypiperidine-1,3-dicarboxylate (302-3) (0.9 g, 2.427 mmol) in methanol (5 mL) and water (5 mL), LiOH (0.581 g, 24.27 mmol) was added at room temperature, and the reaction mixture was cooled to 10° C. for 2 h. 2 The mixture was stirred at room temperature under ambient atmosphere for 5 hours. The pH of the reaction mixture was adjusted to 4 by adding 5M aqueous HCl, extracted with EtOAc, and the combined organic phases were washed with water, brine, dried over sodium sulfate, and concentrated to give the crude product. The crude product was purified by flash chromatography (elution gradient: 0% to 10% MeOH in DCM, 30 min) to give 1-(tert-butoxycarbonyl)-3-(6-chloropyridin-2-yl)-4-hydroxypiperidine-3-carboxylate (302-4). LC-MS: [M+H] + =357.1.

[0362] To a solution of 1-(tert-butoxycarbonyl)-3-(6-chloropyridin-2-yl)-4-hydroxypiperidine-3-carboxylic acid (302-4) (0.8 g, 2.242 mmol) in toluene (20 mL), DPPA (0.926 g, 3.36 mmol) and TEA (1.250 mL, 8.97 mmol) were added at room temperature, and the reaction mixture was cooled to 5° C. for 1 h. 2 After stirring at 100 °C under atmospheric pressure for 3 h, methanol (0.091 mL, 2.242 mmol) was added and the reaction mixture was cooled to 10 °C. 2 The mixture was stirred at 100° C. under ambient atmosphere for 18 h. The reaction mixture was diluted with water, extracted with EtOAc, and the combined organic phases were washed with water, brine, dried over sodium sulfate, and concentrated to give the crude product. The crude product was purified by flash chromatography (elution gradient: 0% to 10% MeOH in DCM, 30 min) to give tert-butyl 3a-(6-chloropyridin-2-yl)-2-oxohexahydrooxazolo[4,5-c]pyridine-5(4H)-carboxylate (302-5). LC-MS: [M+H] + =354.1.

[0363] To a solution of tert-butyl 3a-(6-chloropyridin-2-yl)-2-oxohexahydrooxazolo[4,5-c]pyridine-5(4H)-carboxylate (302-5) (400 mg, 0.113 mmol) in DCM (9 mL) was added TFA (3 mL) at room temperature and the reaction mixture was stirred at room temperature for 3 h. The reaction mixture was concentrated to give crude 3a-(6-chloropyridin-2-yl)hexahydrooxazolo[4,5-c]pyridin-2(3H)-one (302-6). LC-MS: [M+H] + =254.1

[0364] To a solution of methyl 2-(3,4-difluorophenyl)-5-fluoroisonicotinate (Intermediate A) (87 mg, 0.326 mmol) and 3a-(6-chloropyridin-2-yl)hexahydrooxazolo[4,5-c]pyridin-2(3H)-one (302-6) (120 mg, 0.326 mmol) in DMSO (5 mL), DIPEA (5 mL) was added and the reaction mixture was cooled to 5° C. for 1 hour and 10 min. 2The mixture was stirred at 100° C. under ambient atmosphere for 5 hours. The reaction mixture was diluted with water, extracted with EtOAc, and the combined organic phase was washed with water, brine, dried over sodium sulfate, and concentrated to give the crude product. The crude product was purified by flash chromatography (elution gradient: 0% to 10% MeOH in DCM, 30 min) to give methyl 5-(3a-(6-chloropyridin-2-yl)-2-oxohexahydrooxazolo[4,5-c]pyridin-5(4H)-yl)-2-(3,4-difluorophenyl)isonicotinate (302-7). LC-MS: [M+H] + =501.1

[0365] To a solution of methyl 5-(3a-(6-chloropyridin-2-yl)-2-oxohexahydrooxazolo[4,5-c]pyridin-5(4H)-yl)-2-(3,4-difluorophenyl)isonicotinate (302-7) (70 mg, 0.140 mmol) in methanol (5 mL) and THF (5 mL) was added lithium chloride (59.2 mg, 1.398 mmol) and sodium tetrahydroborate (52.9 mg, 1.398 mmol) and the reaction mixture was purged with N 2 The mixture was stirred at room temperature under atmospheric pressure for 5 hours. The reaction mixture was diluted with water and extracted with EtOAc, and the combined organic phase was washed with water, brine, dried over sodium sulfate, and concentrated to give the crude product. The crude product was purified by flash chromatography (elution gradient: 0% to 10% MeOH in DCM, 30 min) to give 3a-(6-chloropyridin-2-yl)-5-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridin-3-yl)hexahydrooxazolo[4,5-c]pyridin-2(3H)-one (302-8). LC-MS: [M+H] + =473.1.

[0366] To a solution of 3a-(6-chloropyridin-2-yl)-5-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridin-3-yl)hexahydrooxazolo[4,5-c]pyridin-2(3H)-one (302-8) (50 mg, 0.106 mmol) in methanol (5 mL) was added a solution of KOH (59.3 mg, 1.057 mmol) in water (5 ml) at room temperature and the reaction mixture was cooled to rt with N 2 The mixture was stirred at 100° C. under atmospheric pressure for 8 hours. The reaction mixture was diluted with water and extracted with EtOAc, and the combined organic phase was washed with water, brine, dried over sodium sulfate, and concentrated to give the crude product. The crude product was purified by flash chromatography (elution gradient: 0% to 10% MeOH in DCM, 30 min) to give 3-amino-3-(6-chloropyridin-2-yl)-1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridin-3-yl)piperidin-4-ol (Intermediate 302-9). LC-MS: [M+H] + =447.1

[0367] Intermediate 303-3: 3-(2,2,2-trifluoroethyl)piperidin-3-amine [ka] Togni reagent (6.42 g, 19.46 mmol) and tetrakis(acetonitrile)copper(I) hexafluorophosphate (483 mg, 1.297 mmol) were dissolved in N 2 The reagent 1 (3 g, 12.97 mmol) and TMSN were then dissolved in DMA (50 mL) under reduced pressure. 3 (2.99 g, 25.9 mmol) was added. The mixture was stirred at room temperature overnight. The mixture was diluted with EA (50 mL) and H 2 The layers were separated and the aqueous layer was extracted with EA (30 ml×3). The combined organic layers were washed with brine (20 ml) and diluted with Na 2 SO 4The filtrate was concentrated and the residue was purified by elution with EA in n-hexane (0-20%) to give benzyl 3-azido-3-(2,2,2-trifluoroethyl)piperidine-1-carboxylate (303-1). 1 H NMR (400 MHz, CDCl 3 7.31-7.51(m,1H),5.03-5.34(m,1H),3.86(d,J=12.80Hz,1H),3.20(d,J=13.55Hz,1 H),2.39(br.s.,1H),1.92-2.01(m,1H),1.81(d,J=9.54Hz,2H)),1.54-1.72(m,1H).

[0368] A solution of benzyl 3-azido-3-(2,2,2-trifluoroethyl)piperidine-1-carboxylate (303-1) (1.3 g, 0.15 mmol) in MeOH (20 ml) was treated with indium powder (1.3 g, 11.4 mmol) and NH 4 Cl (0.609 g, 11.38 mmol) was added. The mixture was heated to reflux in a sealed tube for 2 h. The mixture was cooled to room temperature and filtered. The filtrate was concentrated. The residue was purified by preparative HPLC to give benzyl 3-amino-3-(2,2,2-trifluoroethyl)piperidine-1-carboxylate TFA salt (303-2). LC-MS: [M+H] + =316.9.

[0369] To a solution of benzyl 3-amino-3-(2,2,2-trifluoroethyl)piperidine-1-carboxylate (303-2) (500 mg, 1.58 mmol) in MeOH (40 ml), Pd(OH) 2 (222 mg, 1.581 mmol) 2 The mixture was then added under H 2 Stirred under a balloon for 3 hours. The mixture was filtered and the filtrate was concentrated to 3-(2,2,2-trifluoroethyl)piperidin-3-amine (Intermediate 303-3). 1 H NMR (CDCl 3)δ:3.57-3.35(m,4H),3.02-2.84(m,2H),2.43(q,J=11.1Hz,2H),2.25(dddd,J=1 7.3,13.2,8.6,4.2Hz,1H),1.96(d,J=13.1Hz,1H),1.84-1.59(m,2H)LC-MS:[M+H] + =183.0.

[0370] Intermediate 304-7: (1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridin-3-yl)-3-ethynylpiperidin-3-yl)carbamate [ka] To a solution of 1-benzyl 3-ethyl 3-formylpiperidine-1,3-dicarboxylate (6.5 g, 20.4 mmol, 1.0 equiv.) and dimethyl (1-diazo-2-oxopropyl)phosphonate (7.8 g, 40.8 mmol, 2.0 equiv.) in dry methanol (120 mL) was added K 2 CO 3 (8.5 g, 61.2 mmol, 3.0 equiv.) was added. The mixture was stirred at 1-7 °C for 18 h, filtered, and the filtrate was concentrated. The residue was purified by flash chromatography (PE / EA = 70 / 30) to give 1-benzyl 3-methyl 3-ethynylpiperidine-1,3-dicarboxylate (304-1). 1 H NMR (CDCl 3 400MHz):δ 7.48-7.28(m,5H),5.14(s,2H),4.25-4.09(m,1H),4.00-3.84(m,1H),3.73(s,3H),3.48 (d,J=12.8Hz,1H),3.14-2.97(m,1H),2.20(s,1H),2.06-1.80(m,3H),1.64-1.60(m,1H).

[0371] To a solution of 1-benzyl 3-methyl 3-ethynylpiperidine-1,3-dicarboxylate (Intermediate 304-1) (3.1 g, 10.3 mmol, 1.0 equiv) in THF (50 mL) and water (15 mL) was added LiOH.H 2O (1.7 g, 41.1 mmol, 4.0 equiv) was added. The mixture was stirred at 50° C. for 2 h. The mixture was diluted with water (100 mL) and extracted with EA (80 mL). The aqueous layer was acidified to pH=2 with 1N HCl (aq). The suspension was extracted with DCM (80 mL×2) and Na 2 SO 4 and concentrated to give crude 1-((benzyloxy)carbonyl)-3-ethynylpiperidine-3-carboxylate (304-2). 1 H NMR (CDCl 3 400MHz):δ 10.03(brs,1H),7.40-7.31(m,5H),5.17(s,2H),4.29-4.16(m,1H),4.02-3.88(m,1H),3.49(d,J=1 2Hz, 1H), 3.15-2.98 (m, 1H), 2.24 (s, 1H), 2.12-2.06 (m, 1H), 2.04-1.92 (m, 2H), 1.67-1.64 (m, 1H).

[0372] To a solution of 1-((benzyloxy)carbonyl)-3-ethynylpiperidine-3-carboxylate (304-2) (2.7 g, 9.4 mmol, 1.0 equiv) in DCM (125 mL) was added NH 4 Cl (1 g, 18.8 mmol, 2.0 equiv.), HATU (7.1 g, 18.8 mmol, 2.0 equiv.), and DIEA (4.9 g, 37.6 mmol, 4 equiv.) were added. The reaction mixture was stirred at 25° C. for 16 h. The mixture was washed with water (100 mL) and brine (150 mL) and diluted with Na 2 SO 4 The residue was purified by flash chromatography (PE / EA=50 / 50) to give benzyl 3-carbamoyl-3-ethynylpiperidine-1-carboxylate (304-3). 1 H NMR (400 MHz, CDCl 3):δ 7.35-7.21(m,5H),6.68-6.59(d,J=36.8Hz,1H),5.87(s,1H),5.11-5.01(m,2H),4.25-4.02(m,2H),3 .20-3.14(m,1H),2.89-2.64(m,1H),2.26(d,J=20.8Hz,1H),2.12-1.71(m,3H),1.57(d,J=10Hz,1H).

[0373] To a solution of benzyl 3-carbamoyl-3-ethynylpiperidine-1-carboxylate (304-3) (2.5 g, 8.7 mmol, 1.0 equiv.) in methanol (100 mL) was added KOH (1.2 g, 21.8 mmol, 2.5 equiv.). Then, PhI(OAc) 2 (2.8 g, 8.7 mmol, 1.0 equiv) was added. The resulting mixture was stirred at 3-10 °C for 30 min. The color changed from colorless to yellow. The reaction was concentrated, diluted with water (150 mL), extracted with EA (100 mL × 2), and diluted with Na 2 SO 4 The residue was purified by flash chromatography (PE / EA=70 / 30) to give benzyl 3-ethynyl-3-((methoxycarbonyl)amino)piperidine-1-carboxylate (304-4). 1 H NMR (400MHz, CD 3 OD):δ 7.48-7.30(m,5H),5.20-4.86(m,3H),4.13-3.83(m,2H),3.65(s,3H),3.51-3.22(m,2H),2.69-2.30(m,2H),1.95-1.64(m,3H).

[0374] A mixture of benzyl 3-ethynyl-3-((methoxycarbonyl)amino)piperidine-1-carboxylate (304-4) (800 mg, 2.53 mmol, 1.0 equiv) and TFA (10 mL) in DCM (10 mL) was stirred at 75° C. for 12 h. The mixture was concentrated, diluted with DCM (20 mL) and saturated NaHCO 3 Pour into the solution (50 mL) and add CHCl 3 Extract with / i-PrOH = 3 / 1 (50mL x 4) and Na 2SO 4 and concentrated to give crude methyl (3-ethynylpiperidin-3-yl)carbamate (304-5). LCMS: [M+H] + =182.9.

[0375] To a mixture of methyl (3-ethynylpiperidin-3-yl)carbamate (304-5) (600 mg, 2.53 mmol, 1.0 equiv) and methyl 2-(3,4-difluorophenyl)-5-fluoroisonicotinate (Intermediate A) (670 mg, 2.53 mmol, 1.0 equiv) in DMSO (10 mL) was added DIEA (2.3 g, 7.0 equiv). The reaction mixture was stirred at 120° C. for 2.5 h. The mixture was cooled, diluted with water (100 mL), extracted with EA (50 mL×2), washed with brine (100 mL), and purified by HPLC. 2 SO 4 The mixture was dried at 40° C. and concentrated. The residue was purified by flash chromatography (PE / EA=10 / 1 to 70 / 30) to give methyl 2-(3,4-difluorophenyl)-5-(3-ethynyl-3-((methoxycarbonyl)amino)piperidin-1-yl)isonicotinate (304-6). LCMS: [M+H] + =430.2.

[0376] To a solution of methyl 2-(3,4-difluorophenyl)-5-(3-ethynyl-3-((methoxycarbonyl)amino)piperidin-1-yl)isonicotinate (Intermediate 304-6) (280 mg, 0.652 mmol, 1.0 equiv) in dry THF (12 mL) was added L-selectride (3.9 mL) 2 The solution was stirred at 0° C. for 30 min. The mixture was diluted with saturated NH 4 Quench with Cl solution (30 mL), dilute with water (100 mL), extract with EA (50 mL x 2), and add Na 2 SO 4The mixture was dried at 40° C. and concentrated. The residue was purified by flash chromatography (PE / EA=1 / 1) to give methyl (1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridin-3-yl)-3-ethynylpiperidin-3-yl)carbamate (intermediate 304-7). LCMS: 402.1 [M+H] + =402.1.

[0377] Intermediate 305-5: 3-(difluoromethyl)piperidin-3-amine [ka] N 2 To a solution of 1-benzyl 3-ethyl 3-formylpiperidine-1,3-dicarboxylate (7.0 g, 21.9 mmol, 1.0 equiv) in DCM (100 mL) at 0° C. under ambient conditions was added DAST (14.1 g, 87.8 mmol, 4.0 equiv) dropwise. The reaction was warmed to 10° C. and stirred for 16 h. The reaction was cooled to 0° C. and NaHCO 3 The mixture was carefully quenched with aqueous solution (100 mL). The mixture was extracted with DCM (50 mL×3). The combined organic layers were washed with Na 2 SO 4 The mixture was dried at 40° C., filtered and concentrated in vacuo. The residue was purified by silica gel column (PE / EA=10 / 1) to obtain 4.0 g of crude product, which was further purified by preparative HPLC (0.1% TFA as additive) to obtain 1-benzyl 3-ethyl 3-(difluoromethyl)piperidine-1,3-dicarboxylate 1-benzyl (305-1). 1 H NMR (400 MHz, CDCl 3 ):δ 7.45-7.28(m,5H),5.89(t,J=15.2Hz,1H),5.16(s,2H),4.55-4.37(m,1H),4.27-4.05(m,2H),4.04-3.8 7(m,1H),3.28-3.10(m,1H),3.05-2.90(m,1H),2.25-2.10(m,1H),1.80-1.60(m,3H),1.30-1.12(m,3H).

[0378] THF (10 mL), MeOH (10 mL), and H 2 To a mixture of 1-benzyl 3-ethyl 3-(difluoromethyl)piperidine-1,3-dicarboxylate (305-1) (2.6 g, 7.62 mmol, 1 equiv.) in 2H2O (10 mL) was added LiOH.H 2 2H2O (3.2 g, 76.2 mmol, 10 equiv.) was added. The reaction was stirred at 40° C. for 16 h. The reaction mixture was cooled and acidified to pH=3 with 6 M HCl, then extracted with DCM (30 mL×3). The combined organic layers were washed with brine (50 mL) and diluted with Na 2 SO 4 Drying at 40° C., filtering and concentrating gave 1-((benzyloxy)carbonyl)-3-(difluoromethyl)piperidine-3-carboxylate (305-2). 1 H NMR (400 MHz, CDCl 3 ):δ 7.45-7.25(m,5H),5.93(t,J=15.6Hz,1H),5.17(s,2H),4.55-4.42(m,1H),4.05-3.9 5(m,1H),3.25-3.15(m,1H),3.05-2.95(m,1H),2.28-2.15(m,1H),1.85-1.68(m,3H). LCMS:[M+H] + =314.

[0379] N 2 1-((benzyloxy)carbonyl)-3-(difluoromethyl)piperidine-3-carboxylic acid (305-2) (3.4 g, 10.9 mmol, 1.0 equiv.) in toluene (50 mL) under atmospheric pressure 3 To a mixture of N (2.2 g, 21.72 mmol, 2.0 equiv.) was added DPPA (3.3 g, 11.9 mmol, 1.1 equiv.). The reaction was heated and stirred at 60° C. for 3 h. The reaction mixture was cooled, diluted with EtOAc (100 mL), and then diluted with H 2 The organic layer was washed with O (50 mL × 2). 2 SO 4 Drying at 40° C., filtration and concentration afforded crude benzyl 3-(difluoromethyl)-3-isocyanatopiperidine-1-carboxylate (305-3).

[0380] To a solution of benzyl 3-(difluoromethyl)-3-isocyanatopiperidine-1-carboxylate (305-3) (3.4 g, 10.9 mmol, 1.0 equiv) in dioxane (10 mL) was added 6 M HCl (10 mL). The reaction was stirred at 40° C. for 16 h. The reaction mixture was concentrated to remove dioxane and then diluted with H 2 The mixture was diluted with 20 mL of 2H2O and extracted with EtOAc (20 mL). The aqueous phase was basified with aqueous NaOH to pH = 10. The mixture was extracted with DCM (50 mL x 3). The combined organic layers were washed with Na 2 SO 4 Drying at 40° C., filtration and concentration afforded benzyl 3-amino-3-(difluoromethyl)piperidine-1-carboxylate (305-4). 1 H NMR (400 MHz, CDCl 3 ):δ 7.45-7.28(m,5H),5.56(t,J=16.0Hz,1H),5.15(s,2H),3.88-3.80(m,1H), 3.65-3.50(m,1H),3.35-3.00(m,2H),1.87-1.65(m,4H),1.30-1.15(m,2H). LCMS:[M+H] + =285.

[0381] A mixture of benzyl 3-amino-3-(difluoromethyl)piperidine-1-carboxylate (Intermediate 305-4) (1.8 g, 6.33 mmol, 1 equiv.) and Pd / C (10%, 1.5 g) in EtOH (36 mL) was diluted with H 2 Hydrogenated under a balloon for 3 h. The reaction mixture was filtered and the filtrate was concentrated to dryness to give 3-(difluoromethyl)piperidin-3-amine (Intermediate 305-5). 1 H NMR (400MHz, CD 3 OD):δ 5.68(t,J=16.4Hz,1H),5.15(s,2H),2.88-2.75(m,1H),2.80-2.65(m,1H),2.63-2.52(m,2H),1.80-1.65(m,2H),1.62-1.51(m,2H).

[0382] Intermediate 306-9: tert-Butyl (tert-butoxycarbonyl) (9-((5-((3R,5R)-3-((tert-butoxycarbonyl)amino)-5-(fluoromethyl)piperidin-1-yl)-2-(3-fluoro-4-methoxyphenyl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate [ka] Ethyl 5-bromonicotinate (20.0 g, 86.9 mmol, 1 equiv.), NH 2 Boc (12.2g, 104mmol, 1.2eq.), Xant-Phos (1.60g, 2.77mmol, 0.03eq.), Pd 2 (dba) 3 (1.60 g, 1.75 mmol, 0.02 equiv.), and Cs 2 CO 3 (40.0 g, 123 mmol, 1.4 equiv.), and dioxane (300 mL) were added to a mixture of N 2 The mixture was stirred at 110° C. under atmosphere for 16 hours. The reaction mixture was then concentrated to dryness. Water (800 mL) was added, and the reaction mixture was extracted with EtOAc (300 mL×2). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by combi flash (EtOAc / PE=0-40%) to give ethyl 5-((tert-butoxycarbonyl)amino)nicotinate (306-1). 1 H NMR (400 MHz, CDCl 3 ):δ 8.89(d,J=2.0Hz,1H),8.72(d,J=2.8Hz,1H),8.47(s,1H),6.99(brs,1H),4.41(q,J=7.6Hz,2H),1.54(s,9H),1.40(t,J=7.2Hz,3H).

[0383] To a solution of LAH (3.05 g, 80.3 mmol, 1.6 equiv) in THF (200 mL) was added a solution of ethyl 5-((tert-butoxycarbonyl)amino)nicotinate (306-1) (13.5 g, 50.7 mmol, 1 equiv) in THF (200 mL) dropwise at 0° C. After addition, the reaction mixture was stirred at about 0° C. for 2 h. The reaction mixture was carefully quenched with water (3 mL), 10% NaOH (3 mL), and water (10 mL). The reaction mixture was filtered and the filtrate was concentrated to dryness. The residue was purified by combi Flash (EtOAc / PE=0-100%) to give tert-butyl (5-(hydroxymethyl)pyridin-3-yl)carbamate (306-2). 1 HNMR (400MHz, CDCl 3 ):δ 8.33(d,J=2.4Hz,1H),8.22(d,J=1.6Hz,1H),8.01(s,1H),7.23(brs,1H),4.69(s,2H),1.53(s,9H).

[0384] tert-Butyl (5-(hydroxymethyl)pyridin-3-yl)carbamate (Intermediate 306-2) (10.0 g, 44.6 mmol, 1 equiv.) and PtO 2 Mixture of (3.0 g) with H 2 Hydrogenated under atmosphere (50 psi) at 50° C. for 7 days. The reaction mixture was filtered and the filtrate was concentrated to dryness to give tert-butyl (5-(hydroxymethyl)piperidin-3-yl)carbamate (306-3). LC-MS: [M+H] + =230.1

[0385] THF (100 mL) and H 2 tert-Butyl (5-(hydroxymethyl)piperidin-3-yl)carbamate (306-3) (10.0 g, impure, 1 equiv.) and NaHCO in 2H2O (100 mL). 3To a suspension of (10.9 g, 130 mmol, 3 equiv.) was added CbzCl (8.20 g, 47.4 mmol, 1.1 equiv.) dropwise at about 0° C. After addition, the reaction mixture was stirred at about 20° C. for 16 h. The reaction mixture was poured into water (400 mL) and extracted with EtOAc (200 mL×3). The combined organic layers were washed with Na 2 SO 4 The residue was purified by combi Flash (EtOAc / PE=0-70%) to give benzyl 3-((tert-butoxycarbonyl)amino)-5-(hydroxymethyl)piperidine-1-carboxylate benzyl (306-4). 1 H NMR (400 MHz, CDCl 3 ):δ 7.40-7.25(m,5H),5.25-5.05(m,2H),4.80-4.10(m,3H),3.65-3.40(m,4H),2.60- 2.45(m,1H),2.15-2.00(m,1H),1.98-1.65(m,2H),1.43(s,9H),1.15-1.00(m,1H). LC-MS:[M+Na] + =387.1.

[0386] To a mixture of benzyl 3-((tert-butoxycarbonyl)amino)-5-(hydroxymethyl)piperidine-1-carboxylate (306-4) (3.50 g, 9.60 mmol, 1.0 equiv) in THF (60 mL) was added Et 3 N (13.6g, 134mmol, 14.0eq), FSO 2 (CF 2 ) 3 CF 3 (5.80 g, 19.2 mmol, 2.0 equiv.), and TEA.3HF (6.19 g, 38.4 mmol, 4.0 equiv.) were added. The reaction mixture was stirred at 25° C. for 12 h. The reaction mixture was concentrated in vacuo. The residue was purified by silica gel column chromatography (eluent: PE / EA10 / 1-3 / 1) to give benzyl 3-((tert-butoxycarbonyl)amino)-5-(fluoromethyl)piperidine-1-carboxylate (306-5). LC-MS: [M+Na] + =389.1.

[0387] To a solution of benzyl 3-((tert-butoxycarbonyl)amino)-5-(fluoromethyl)piperidine-1-carboxylate (306-5) (2.50 g, 6.83 mmol, 1.0 equiv) in MeOH (30 mL) was added Pd(OH) 2 / C (250 mg, 10 wt%) was added. The reaction mixture was stirred at 25° C. for 1 h. The mixture was filtered and the filtrate was concentrated in vacuo to give tert-butyl (5-(fluoromethyl)piperidin-3-yl)carbamate (305-6). LC-MS: [M+H] + =233.1.

[0388] To a solution of tert-butyl (5-(fluoromethyl)piperidin-3-yl)carbamate (306-6) (1.50 g, 6.51 mmol, 1.0 equiv.) in DMSO (15 mL) was added DIEA (5.89 g, 45.6 mmol, 7.0 equiv.) and methyl 5-fluoro-2-(3-fluoro-4-methoxyphenyl)isonicotinate (Intermediate D) (1.82 g, 6.51 mmol, 1.0 equiv.). The reaction mixture was stirred at RT for 2 h at 4° C. for 1 h at 4° C. for 2 h at 4° C. 2 The mixture was stirred at 120° C. for 3 h under reduced pressure. The reaction mixture was diluted with water (100 mL) and extracted with EtOAc (20 mL×3). The combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography (PE / EA10 / 1-2 / 1) and further purified by preparative HPLC (0.05% NH as an additive). 3 H 2 0) to give methyl 5-((3R,5R)-3-((tert-butoxycarbonyl)amino)-5-(fluoromethyl)piperidin-1-yl)-2-(3-fluoro-4-methoxyphenyl)isonicotinate (306-7). 1 H NMR (400MHz, CD 3OD):δ 8.45(s,1H),7.93(s,1H),7.76-7.67(m,2H),7.19(t,J=8.4Hz,1H),4.51 (d,J=6.0Hz,1H),4.39(d,J=6.0Hz,1H),4.02(s,3H),3.94(s,3H),3.92-3 .86(m,1H),3.35-3.24(m,1H),3.20-3.08(m,2H),3.01-2.92(m,1H),2.4 6-2.31(m,1H),1.84-1.76(m,1H),1.64-1.54(m,1H),1.42-1.41(m,10H). LC-MS:[M+H] + =492.2

[0389] To a mixture of methyl 5-((3R,5R)-3-((tert-butoxycarbonyl)amino)-5-(fluoromethyl)piperidin-1-yl)-2-(3-fluoro-4-methoxyphenyl)isonicotinate (306-7) (120 mg, 0.24 mmol, 1.0 equiv.) and LiCl (102 mg, 2.44 mmol, 10 equiv.) in THF (2 mL) and MeOH (2 mL) was added NaBH 4 (181 mg, 4.88 mmol, 20 equiv) was added in portions. After the addition, the mixture was stirred at 25° C. for 1 h. Then, water (20 mL) was added and the mixture was extracted with EtOAc (10 mL×3). The combined organic phase was washed with brine (10 mL×2) and anhydrous Na 2 SO 4 Drying at 40° C., filtering and concentrating in vacuo gave tert-butyl ((3R,5R)-1-(6-(3-fluoro-4-methoxyphenyl)-4-(hydroxymethyl)pyridin-3-yl)-5-(fluoromethyl)piperidin-3-yl)carbamate (306-8). LC-MS: [M+H] + =464.3

[0390] tert-Butyl ((3R,5R)-1-(6-(3-fluoro-4-methoxyphenyl)-4-(hydroxymethyl)pyridin-3-yl)-5-(fluoromethyl)piperidin-3-yl)carbamate (Intermediate 306-8) (100 mg, 0.22 mmol, 1.0 equiv., Intermediate B (109 mg, 0.32 mmol, 1.5 equiv.), n-Bu 3 To a mixture of P (66.0 mg, 0.32 mmol, 1.5 equiv.) and THF (10 mL) was added DIAD (66.0 mg, 0.32 mmol, 1.5 equiv.) dropwise. After addition, the mixture was rinsed with N 2 The mixture was stirred under reduced pressure at 25° C. for 12 hours. The mixture was concentrated in vacuo. The residue was purified by silica gel column (PE / EA=5:1 to 1:1) to give tert-butyl (tert-butoxycarbonyl(9-((5-((3R,5R)-3-(tert-butoxycarbonyl)amino)-5-(fluoromethyl)piperidin-1-yl)-2-(3-fluoro-4-methoxyphenyl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate (intermediate 306-9). LC-MS: [M+Na] + =803.4

[0391] Intermediate 311-11: Methyl 5-(3-(benzyl(methoxycarbonyl)amino)-2-((difluoromethoxy)methyl)piperidin-1-yl)-2-(3,4-difluorophenyl)isonicotinate [ka] To a solution of 3-aminopicolinic acid (50 g, 360 mmol, 1.0 equiv) in anhydrous MeOH (500 mL), add concentrated H 2 SO 4 (106 g, 1090 mmol, 3.0 equiv.) was added dropwise. The reaction was stirred at 80 °C for 48 h. SOCl 2 (130 g, 1090 mmol, 3.0 equiv) was added to the reaction and stirred at 80° C. for an additional 48 h. The reaction was concentrated, diluted with water (500 mL) and K 2 CO 3 Basify with (s) to pH=9, extract with EA (500mL×3), wash with brine (500mL), and add Na2 SO 4 and concentrated to give methyl 3-aminopicolinate (311-1). 1 H NMR (400 MHz, CDCl 3 )δ=8.04(d,J=4.3Hz,1H),7.20(dd,J=4.3,8.4Hz,1H),7.04(dd,J=1.3,8.4Hz,1H),5.76(br s,2H),3.98-3.92(m,3H).

[0392] Methyl 3-aminopicolinate (311-1) (27 g, 180 mmol, 1.0 equiv.) and K 2 CO 3 To a solution of (48 g, 350 mmol, 2.0 equiv) was added methyl carbonochloridate (33 g, 350 mmol, 2.0 equiv) at 20 °C for 18 h. The reaction mixture was diluted with water (500 mL) and concentrated. The residue was acidified with 1N HCl to PH = 3, extracted with EA (500 mL x 3) and washed with saturated NaHCO 3 (500 mL) and brine (300 mL), and then washed with Na 2 SO 4 and concentrated to give methyl 3-((methoxycarbonyl)amino)picolinate (311-2). 1 H NMR (400 MHz, CDCl 3 )δ=10.39(br s,1H),8.82(dd,J=1.4,8.7Hz,1H),8.34(dd,J=1.5,4.4Hz,1H),7.51-7.40(m,1H),3.99(s,3H),3.78(s,3H).

[0393] Methyl 3-((methoxycarbonyl)amino)picolinate (Intermediate 311-2) (22 g, 105 mmol, 1.0 equiv.) in AcOH (220 mL) and PtO 2 (5 g, 23 wt%) solution in H 2 The mixture was stirred under 25° C. and 5 MPa for 144 h. The reaction mixture was filtered and concentrated to give methyl (2S,3R)-3-((methoxycarbonyl)amino)piperidine-2-carboxylate (311-3).

[0394] To a solution of methyl (2S,3R)-3-((methoxycarbonyl)amino)piperidine-2-carboxylate (311-3) (30 g, ca. 105 mmol, 1.0 equiv.) in THF (300 mL) was added TEA (53 g, 525 mmol, 5.0 equiv.) and Boc 2 2H2O (69 g, 315 mmol, 3.0 equiv) was added at 20° C. for 18 h. The reaction mixture was concentrated, diluted with water (100 mL), acidified with 1N HCl to PH=3, extracted with EA (100 mL×3), washed with brine (100 mL), and diluted with Na 2 SO 4 The residue was purified by combi-flash (EA%=0%-50%) to give 1-(tert-butyl) 2-methyl(2S,3R)-3-((methoxycarbonyl)amino)piperidine-1,2-dicarboxylate (311-4).

[0395] To a solution of 1-(tert-butyl) 2-methyl(2S,3R)-3-((methoxycarbonyl)amino)piperidine-1,2-dicarboxylate (311-4) (33 g, 104 mmol, 1.0 equiv.) in anhydrous THF (660 mL) was added LiAlH 4 (5.15 g, 136 mmol, 1.3 equiv.) was added at -78°C to -10°C for 2 h. The reaction mixture was diluted with saturated NH 4 Cl (700 mL), acidified with 1N HCl to PH=5, extracted with EA (500 mL×3), washed with brine (500 mL), and added Na 2 SO 4 The residue was purified by combi-flash (EA%=0%-70%) to give tert-butyl (2S,3R)-2-(hydroxymethyl)-3-((methoxycarbonyl)amino)piperidine-1-carboxylate (311-5). 1 H NMR (400 MHz, CDCl 3)δ:5.70(br s,1H),4.44(br d,J=5.5Hz,1H),3.92(br d,J=10.9Hz,2H),3.86-3.71(m,2H),3.64(br s, 3H), 2.98-2.64 (m, 2H), 1.92-1.80 (m, 1H), 1.73-1.63 (m, 1H), 1.61-1.50 (m, 2H), 1.44 (s, 9H). LC-MS:[M+H] + =289.2.

[0396] To a mixture of tert-butyl (2S,3R)-2-(hydroxymethyl)-3-((methoxycarbonyl)amino)piperidine-1-carboxylate (311-5) (5 g, 17 mmol, 1.0 equiv), BnBr (12 g, 69 mmol, 1.0 equiv), and TBAI (3.1 g, 9 mmol, 0.5 equiv) in anhydrous THF (50 mL) was added NaH (2.6 g, 65 mmol, 3.8 equiv) at 20 °C. After the addition, the reaction was stirred at 60 °C for 2 h. The reaction mixture was diluted with saturated NH 4 Pour into a solution of Cl (200 mL), extract with EA (100 mL × 3), wash with brine (50 mL), and add Na 2 SO 4 The residue was purified by combi-flash (EA%=0%-50%) to give tert-butyl (2S,3R)-3-(benzyl(methoxycarbonyl)amino)-2-((benzyloxy)methyl)piperidine-1-carboxylate (311-6). 1 H NMR (400 MHz, CDCl 3 )δ:7.53-6.90(m,10H),4.90-3.31(m,12H),3.08-2.74(m,1H),1.97-1.79(m,1H),1.62-1.50(m,1H),1.50-1.42(m,1H),1.41-1.28(m,9H). LC-MS:[M+H] + =469.4.

[0397] A solution of tert-butyl (2S,3R)-3-(benzyl(methoxycarbonyl)amino)-2-((benzyloxy)methyl)piperidine-1-carboxylate (Intermediate 311-6) (7.2 g, 15 mmol, 1.0 equiv) in DCM (40 mL) and TFA (20 mL) was stirred at 20° C. for 20 h. The reaction mixture was stirred at 20° C. for 20 h. 2 Dilute with HO (100 mL) and K 2 CO 3 Basify with (s) to pH=9, extract with EA (100mL×3), wash with brine, and add Na 2 SO 4 Drying at 40° C. and concentration gave methyl benzyl ((2S,3R)-2-((benzyloxy)methyl)piperidin-3-yl)carbamate (311-7). LC-MS: [M+H] + =369.5.

[0398] CDCl 3 A mixture of methyl 2-(3,4-difluorophenyl)-5-fluoroisonicotinate (Intermediate A) (30 g, 0.112 mmol, 1.0 equiv) and m-CPBA (30 g, 0.174 mmol, 1.55 equiv) in 100 mL of ethyl acetate was stirred at 65° C. for 20 h. Additional m-CPBA (30 g, 0.174 mmol, 1.55 equiv) was added to the reaction mixture and the reaction was stirred at 65° C. for 72 h. The reaction mixture was diluted with water (300 mL) and diluted with K 2 CO 3 Basify with (s) to pH=8, extract with EA (300mL×3), wash with brine (300mL), and add Na 2 SO 4 The crude was purified by trituration with PE / MTBE=1 / 1 (100 mL) to give 2-(3,4-difluorophenyl)-5-fluoro-4-(methoxycarbonyl)pyridine 1-oxide (311-7A). LC-MS: [M+H] + =284.2.

[0399] A mixture of methyl benzyl ((2S,3R)-2-((benzyloxy)methyl)piperidin-3-yl)carbamate (311-7) (17 g, 0.06 mol, 4.0 equiv.) and 2-(3,4-difluorophenyl)-5-fluoro-4-(methoxycarbonyl)pyridine 1-oxide (311-7A) (6 g, ca. 15 mmol, 1.0 equiv.) in DIPEA (40 mL) and DMSO (40 mL) was stirred at 120° C. for 5 h. 2 Dilute with 200 mL of HO (100 mL), extract with EA (50 mL × 3), wash with brine (20 mL), and 2 SO 4 The residue was purified by combi-flash (EA%=10%-100%) to give 5-(3-(benzyl(methoxycarbonyl)amino)-2-((benzyloxy)methyl)piperidin-1-yl)-2-(3,4-difluorophenyl)-4-(methoxycarbonyl)pyridine 1-oxide (311-8). LCMS: [M+H] + =632.5.

[0400] 5-(3-(benzyl(methoxycarbonyl)amino)-2-((benzyloxy)methyl)piperidin-1-yl)-2-(3,4-difluorophenyl)-4-(methoxycarbonyl)pyridine 1-oxide (311-8) (5200 mg, ca. 4.69 mmol, 1.0 equiv.) and Pd(OH) in MeOH (25 mL) and EA (25 mL). 2 A mixture of (2000 mg, 38 wt%) was stirred at 60° C. and 50 Psi for 20 h. The reaction mixture was filtered and concentrated. The residue was purified by combi-flash (EA%=0%-60%) to give methyl 5-(3-(benzyl(methoxycarbonyl)amino)-2-((benzyloxy)methyl)piperidin-1-yl)-2-(3,4-difluorophenyl)isonicotinate (311-9). LCMS: [M+H] + =616.7.

[0401] Methyl 5-(3-(benzyl(methoxycarbonyl)amino)-2-((benzyloxy)methyl)piperidin-1-yl)-2-(3,4-difluorophenyl)isonicotinate (311-9) (1500 mg, 2.44 mmol, 1.0 equiv) and Pd(OH) in EA (45 mL). 2 A mixture of (1500 mg, 100 wt%) was stirred at 60° C. and 50 Psi for 18 h. The reaction mixture was filtered and concentrated. The residue was purified by combi-flash (EA%=0%-50%) to give methyl 5-(3-(benzyl(methoxycarbonyl)amino)-2-(hydroxymethyl)piperidin-1-yl)-2-(3,4-difluorophenyl)isonicotinate (311-10). LCMS: [M+H] + =526.5.

[0402] To a solution of methyl 5-(3-(benzyl(methoxycarbonyl)amino)-2-(hydroxymethyl)piperidin-1-yl)-2-(3,4-difluorophenyl)isonicotinate (311-10) (300 mg, 0.571 mmol) and CuI (21.74 mg, 0.114 mmol) in MeCN (10 mL) was added 2,2-difluoro-2-(fluorosulfonyl)acetic acid (0.113 mL, 1.142 mmol) dropwise at 55° C. and the reaction mixture was stirred at 55° C. for 2 h. The reaction mixture was diluted with EA and washed with water and brine. The organic layer was extracted with Na 2 SO 4 The crude product was purified by flash chromatography (elution gradient: 0% to 40% EtOAc in PE, 30 min) to give methyl 5-(3-(benzyl(methoxycarbonyl)amino)-2-((difluoromethoxy)methyl)piperidin-1-yl)-2-(3,4-difluorophenyl)isonicotinate (Intermediate 311-11). LCMS: [M+H] + =576.3. EXAMPLES

[0403] Example 1: (R)-9-((5-(3-aminopiperidin-1-yl)-2-(3-fluoro-4-methoxyphenyl)pyridin-4-yl)methyl)-9H-purin-6-amine [ka] A suspension of 5-fluoro-2-(3-fluoro-4-methoxyphenyl)isonicotinaldehyde (Intermediate 1-1) (800 mg, 3.21 mmol), (R)-tert-butyl piperidin-3-ylcarbamate (643 mg, 3.21 mmol), and potassium carbonate (887 mg, 6.42 mmol) in DMF (15 mL) was stirred at 100° C. for 20 h. The mixture was cooled to 100° C. and cooled to 37° C. for 20 h. 2 The mixture was diluted with 20 mL of H2O (30 mL) and extracted with EtOAc (20 mL x 3). The combined organic layers were 2 Wash with 20 mL of O (20 mL × 2) and saline (20 mL), and then with anhydrous Na 2 SO 4 The residue was purified by silica gel column chromatography (0-30% EtOAc in PE, 30 min) to give tert-butyl (R)-(1-(6-(3-fluoro-4-methoxyphenyl)-4-formylpyridin-3-yl)piperidin-3-yl)carbamate (1-2). 1H NMR(400MHz,DMSO-d6)δ 10.24(s,1H),8.63(s,1H),8.01(s,1H),7.93-7.83(m,2H),7.27(t,J=8.8Hz,1H),7.04(d,J=7.5Hz,1H),3.90(s,3H),3.63(s,1H),3.42-3. 37(m,1H),3.30-3.23(m,1H),3.06-2.94(m,1H),2.84(dd,J=11.7,9.0Hz,1H),1.85(d,J=11.2Hz,2H),1.70(d,J=10.5Hz,1H),1.40(s,10H). LC-MS:[M+H] + =429.9.

[0404] To a suspension of tert-butyl (R)-(1-(6-(3-fluoro-4-methoxyphenyl)-4-formpyridin-3-yl)piperidin-3-yl)carbamate (1-2) (260 mg, 0.605 mmol) in MeOH (15 mL) was added sodium borohydride (22.9 mg, 0.605 mmol). The mixture was stirred at room temperature for 30 min and the solvent was removed under reduced pressure. The residue was redissolved in DCM (30 mL), worked up under aqueous conditions and concentrated. The residue was purified by flash chromatography (10-50% EtOAc in PE, 30 min) to give tert-butyl (R)-(1-(6-(3-fluoro-4-methoxyphenyl)-4-(hydroxymethyl)pyridin-3-yl)piperidin-3-yl)carbamate (1-3). 1 H NMR (400MHz, CD 3 OD)δ 8.26(s,1H),7.93(s,1H),7.75-7.62(m,2H),7.21(t,J=8.5Hz,1H),5.52(s, 1H),4.88-4.72(m,2H),3.95(s,3H),3.71(dt,J=9.5,4.8Hz,1H),3.21(dd,J =11.2,3.7Hz,1H),3.07(d,J=11.6Hz,1H),2.83(t,J=10.7Hz,1H),2.66(t,J =10.0Hz,1H),2.06-1.86(m,2H),1.78(q,J=13.1,11.6Hz,1H),1.47(s,9H). LC-MS: [M+H] + =431.9.

[0405] A mixture of tert-butyl (R)-(1-(6-(3-fluoro-4-methoxyphenyl)-4-(hydroxymethyl)pyridin-3-yl)piperidin-3-yl)carbamate (1-3) (370 mg, 0.857 mmol), intermediate B (300 mg, 0.895 mmol), and PPh 3 (704 mg, 2.68 mmol) was added dropwise to a solution of DEAD (0.425 mL, 2.68 mmol) at 0° C., and the mixture was stirred at room temperature for 1 h. 2The mixture was diluted with 20 mL of 2H2O and extracted with EtOAc (20 mL x 3). The combined organic layers were washed with brine (20 mL) and 2 SO 4 The residue was purified by silica gel column chromatography (10-50% EtOAc in hexanes, 40 min) to give tert-butyl (R)-(tert-butoxycarbonyl)(9-((5-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-2-(3-fluoro-4-methoxyphenyl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate (1-4). 1 H NMR (400MHz, CD 3 OD)δ 8.88(s,1H),8.76(s,1H),8.45(s,1H),7.54-7.44(m,2H),7.26(s,1H),7.13(t,J =8.6Hz,1H),5.90-5.72(m,2H),3.90(s,3H),3.85(d,J=4.6Hz,1H),3.42-3.37(m, 1H),3.18-3.09(m,1H),2.99(t,J=10.1Hz,1H),2.88(d,J=10.1Hz,1H),1.97(d,J= 10.2Hz, 2H), 1.82 (d, J=6.8Hz, 1H), 1.64-1.54 (m, 1H), 1.44 (s, 9H), 1.36 (s, 18H). LC-MS:[M+H] + =749.0.

[0406] To a solution of tert-butyl (R)-(tert-butoxycarbonyl)(9-((5-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-2-(3-fluoro-4-methoxyphenyl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate (1-4) (360 mg, 0.481 mmol) in DCM (9 mL) was added TFA (3 mL). The mixture was stirred at room temperature for 1 h. The mixture was concentrated in vacuo. The residue was purified by preparative HPLC (basic conditions, NH 3 H 2 O% = 0.05%, MECN / H 2O=0-95%, 12 min) to give (R)-9-((5-(3-aminopiperidin-1-yl)-2-(3-fluoro-4-methoxyphenyl)pyridin-4-yl)methyl)-9H-purin-6-amine (Example 1). 1 H NMR (400MHz, CD 3 OD)δ 8.59-8.32(m,3H),7.69-7.53(m,2H),7.37(s,1H),7.17(td,J=8.6,1.7Hz,1H),5.85-5.63(m,2H),3.93(d,J=1.8Hz,3H),3.62(s,1H),3.56 -3.44(m,1H),3.19-3.07(m,2H),3.04(d,J=9.5Hz,1H),2.22-2.12(m,1H),2.04(s,1H),1.89(dd,J=9.0,4.2Hz,1H),1.76(d,J=10.1Hz,1H). LC-MS:[M+H] + =449.1.

[0407] Following procedures similar to those described in Example 1, Examples 2-21 can be prepared. [Table 4-1] [Table 4-2] [Table 4-3] [Table 4-4] [Table 4-5] [Table 4-6] [Table 4-7]

[0408] Example 22: (R)-9-((5-(3-amino-3-(cyclopropoxymethyl)piperidin-1-yl)-2-(3,4-difluorophenyl)pyridin-4-yl)methyl)-9H-purin-6-amine [ka] A mixture of tert-butyl (3-(cyclopropoxymethyl)piperidin-3-yl)carbamate (Intermediate 22-7) (300 mg, 1.11 mmol, 1.0 equiv), methyl 2-(3,4-difluorophenyl)-5-fluoroisonicotinate (Intermediate A) (296 mg, 1.11 mmol, 1.0 equiv), and DIPEA (1.00 g, 7.77 mmol, 7.0 equiv) in DMSO (3 mL) was cooled to 10° C. for 2 h. 2 The mixture was stirred at 120° C. for 3 h under reduced pressure. The mixture was quenched with water (200 mL) at 10° C. and extracted with EtOAc (100 mL×3). The combined organic layer was washed with anhydrous Na 2 SO 4 The residue was purified by Combi Flash (20% EtOAc in PE) to give methyl 5-(3-((tert-butoxycarbonyl)amino)-3-(cyclopropoxymethyl)piperidin-1-yl)-2-(3,4-difluorophenyl)isonicotinate (22-8). 1 H NMR (400 MHz, CDCl 3 )δ ppm 8.48(1H,s),7.86(1H,s),7.83(1H,dd,J=10.0,2.0Hz),7.67(1H,d,J=8.4Hz) ,7.23(1H,dd,J=8.4,1.6Hz),5.63(1H,brs),3.80-4.05(4H,m),3.57(1H,d,J =9.6Hz),3.15-3.35(3H,m),2.80-2.95(2H,m),2.30-2.45(1H,m),1.85-2.00 (1H,m),1.65-1.75(1H,m),1.46(9H,s),1.25-1.40(1H,m),0.40-0.60(4H,m).

[0409] To a solution of methyl 5-(3-((tert-butoxycarbonyl)amino)-3-(cyclopropoxymethyl)piperidin-1-yl)-2-(3,4-difluorophenyl)isonicotinate (22-8) (330 mg, 0.638 mmol, 1.0 equiv) in THF / MeOH (10 mL, v / v=1 / 1) was added LiCl (270 mg, 6.38 mmol, 10.0 equiv), followed by NaBH 4 (480 mg, 12.75 mmol, 20.0 equiv) was added at 15-20 °C. The resulting mixture was stirred at 30 °C for 2 h. The reaction was quenched with water (100 mL) and extracted with EtOAc (100 mL × 3). The combined organic layer was washed with anhydrous Na 2 SO 4 and concentrated in vacuo to give crude tert-butyl (3-(cyclopropoxymethyl)-1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridin-3-yl)piperidin-3-yl)carbamate (22-9). 1 H NMR (400 MHz, CDCl 3 )δ ppm 8.43(1H,s),7.75-7.85(1H,m),7.60-7.70(2H,m),7.15-7.25(1H,m),4.79 (2H,s),4.76(1H,brs),3.65-3.85(3H,m),3.35-3.45(1H,m),3.25-3.35(1H ,m),3.05-3.15(1H,m),2.80-2.95(2H,m),2.05-2.15(1H,m),1.85-1.95(1 H,m),1.65-1.75(1H,m),1.55-1.60(1H,m),1.44(9H,s),0.40-0.60(4H,m).

[0410] A solution of tert-butyl (3-(cyclopropoxymethyl)-1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridin-3-yl)piperidin-3-yl)carbamate (22-9) (300 mg, 0.613 mmol, 1.0 equiv.), intermediate B (310 mg, 0.919 mmol, 1.5 equiv.), and Bu in THF (10 mL). 3To a solution of P (372 mg, 1.84 mmol, 3.0 equiv) was added DIAD (372 mg, 1.84 mmol, 3.0 equiv) at 0 °C. The mixture was stirred at 20-30 °C for 2 h. The reaction was concentrated in vacuo. The residue was purified by Combi Flash (30-40% EtOAc in PE) to give tert-butyl (tert-butoxycarbonyl) (9-((5-(3-(tert-butoxycarbonyl)amino)-3-(cyclopropoxymethyl)piperidin-1-yl)-2-(3,4-difluorophenyl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate (22-10). 1 H NMR (400 MHz, CDCl 3 )δ ppm 8.88(1H,s),8.52(1H,s),8.25(1H,s),7.60-7.70(1H,m),7.35-7.45(1H,m),7.10-7.25(2H, m),5.70(1H,d,J=15.6Hz),5.53(1H,d,J=15.6Hz),5.19(1H,brs),3.75-3.90(2H,m),3.40-3 intermediate.50(1H,m),3.25-3.35(1H,m),2.90-3.10(3H,m),2.15-2.30(1H,m),1.90-2.0 0(1H,m),1.70-1.80(1H,m),1.55-1.65(1H,m),1.45(18H,s),1.35(9H,s),0.40-0.60(4H,m).

[0411] A solution of tert-butyl (tert-butoxycarbonyl)(9-((5-(3-((tert-butoxycarbonyl)amino)-3-(cyclopropoxymethyl)piperidin-1-yl)-2-(3,4-difluorophenyl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate (22-10) (270 mg, 0.335 mmol) in 20% TFA in DCM (7.5 mL) was stirred at 15-20 °C for 2 h. The pH of the reaction was then adjusted to 0.05 with NH 3 H 2 The mixture was adjusted to 8-9 with O and extracted with DCM (100 mL x 3). The combined organic layer was washed with anhydrous Na 2 SO 4The residue was purified by preparative HPLC (0.1% NH 3 H 2 03C) and purification by SFC to give (R)-9-((5-(3-amino-3-(cyclopropoxymethyl)piperidin-1-yl)-2-(3,4-difluorophenyl)pyridin-4-yl)methyl)-9H-purin-6-amine (Example 22). 1 H NMR (400MHz, CD 3 OD)δ ppm 8.51(1H,s),8.25(1H,d,J=4.0Hz),7.70-7.80(1H,m),7.50-7.60(1H,m),7.25-7.40(2H,m),5.66(2H,s),3.60-3. 80(1H,m),3.45-3.55(1H,d,J=9.6Hz),2.90-3.15(4H,m),1.55-2.00(4H,m),1.25-1.40(1H,m),0.35-0.65(4H,m). LC-MS:[M+H] + =506.8.

[0412] Examples 23-46 can be prepared following procedures similar to those described in Example 22. [Table 5-1] [Table 5-2] [Table 5-3] [Table 5-4] [Table 5-5] [Table 5-6] [Table 5-7]

[0413] Example 47: (S)-9-((5-(3-amino-3-(2,2-difluoroethyl)piperidin-1-yl)-2-(3,4-difluorophenyl)pyridin-4-yl)methyl)-9H-purin-6-amine [ka]

[0414] To a solution of methyl 2-(3,4-difluorophenyl)-5-fluoroisonicotinate (Intermediate A) (1.35 g, 5.05 mmol) and tert-butyl (3-(2,2-difluoroethyl)piperidin-3-yl)carbamate (Intermediate 47-6) (1.3 g, 5.05 mmol) in DMSO (30 mL), DIPEA (20 mL) was added at room temperature and the reaction mixture was cooled to 5° C. 2 The mixture was stirred at 120° C. under atmosphere for 8 hours. The reaction mixture was diluted with water and extracted with EtOAc (20 mL×3), and the combined organic phase was washed with water (20 mL), brine (20 mL), dried over anhydrous sodium sulfate, and concentrated to give the crude product. The crude product was purified by flash chromatography (elution gradient: 0% to 30% EtOAc in PE, 30 min) to give methyl 5-(3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoroethyl)piperidin-1-yl)-2-(3,4-difluorophenyl)isonicotinate (47-7). LC-MS: [M+H] + =512.0.

[0415] To a solution of methyl 5-(3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoroethyl)piperidin-1-yl)-2-(3,4-difluorophenyl)isonicotinate (47-7) (1.2 g, 2.346 mmol) in THF (30 mL) and methanol (30 mL), lithium chloride (1.97 g, 46.9 mmol) and sodium tetrahydroborate (1.78 g, 46.9 mmol) were added at room temperature, and the reaction mixture was diluted with N 2The mixture was stirred at room temperature under atmosphere for 5 hours. The reaction mixture was diluted with water (20 mL), extracted with EtOAc (20 mL×3), and the combined organic phase was washed with water (20 mL), brine (20 mL), dried over anhydrous sodium sulfate, and concentrated to give a residue. The residue was purified by flash chromatography (elution gradient: 10% to 50% EtOAc in hexanes, 30 min) to give tert-butyl (3-(2,2-difluoroethyl)-1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridin-3-yl)piperidin-3-yl)carbamate (47-8). LC-MS: [M+H] + =484.2.

[0416] To a solution of tert-butyl (3-(2,2-difluoroethyl)-1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridin-3-yl)piperidin-3-yl)carbamate (47-8) (865 mg, 1.789 mmol), Intermediate B (600 mg, 1.789 mmol), and triphenylphosphine (939 mg, 3.58 mmol) in THF (20 mL) was added DEAD (0.567 mL, 3.58 mmol) dropwise at 0 °C and the reaction mixture was quenched with N 2 The mixture was stirred at 0° C. under atmosphere for 0.5 h. The reaction mixture was diluted with water (20 mL) and extracted with EtOAc (20 mL×3), and the combined organic phase was washed with water (10 mL), brine (20 mL), dried over sodium sulfate, and concentrated to give the crude product. The crude product was purified by flash chromatography (elution gradient: 0% to 10% MeOH in DCM, 30 min) to give tert-butyl (tert-butoxycarbonyl) (9-(5-(3-(tert-butoxycarbonyl)amino)-3-(2,2-difluoroethyl)piperidin-1-yl)-2-(3,4-difluorophenyl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate (47-9). LC-MS: [M+H] + =801.1.

[0417] To a solution of tert-butyl (tert-butoxycarbonyl)(9-((5-(3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoroethyl)piperidin-1-yl)-2-(3,4-difluorophenyl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate (47-9) (700 mg, 0.875 mmol) in DCM (18 mL) was added TFA (6 mL) and the reaction mixture was cooled to 10° C. with N 2 The mixture was stirred at room temperature under atmospheric pressure for 2 hours. The reaction mixture was concentrated in vacuo to give the crude product. The crude product was purified by preparative HPLC (basic conditions, NH 3 H 2 O% = 0.05%, MECN / H 2 O = 0-95%, 12 min) to give the pure racemic product, which was isolated by SFC to give (S)-9-((5-(3-amino-3-(2,2-difluoroethyl)piperidin-1-yl)-2-(3,4-difluorophenyl)pyridin-4-yl)methyl)-9H-purin-6-amine (Example 47). 1 H NMR (400MHz, CD 3 OD)δ 8.49(s,1H),8.23(d,J=3.9Hz,2H),7.75(ddd,J=12.2,7.8,2.2Hz,1H),7.53(s,1H),7.34-7.22(m,2H),6.38-6.02(m,1H),5.73 -5.52(m,2H),3.00(d,J=9.4Hz,3H),2.92(d,J=10.5Hz,1H),2.11(q,J=18.5,18.0Hz,2H),1.92(s,1H),1.73(d,J=28.9Hz,3H). LC-MS:[M+H] + =501.3.

[0418] Examples 48-77 can be prepared from the corresponding intermediates following procedures similar to those described in Example 47. [Table 6-1] [Table 6-2] [Table 6-3] [Table 6-4] [Table 6-5] [Table 6-6] [Table 6-7] [Table 6-8] [Table 6-9]

[0419] Example 78: (R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(3-fluorophenyl)pyridin-3-yl)-N-methylpiperidine-3-carboxamide [ka]

[0420] To a solution of methyl tert-butyl 3-(methylcarbamoyl)piperidin-3-ylcarbamate (intermediate 78-5) (200 mg, 0.777 mmol), methyl 5-fluoro-2-(3-fluorophenyl)isonicotinate (intermediate C) (161 mg, 0.648 mmol) in DMSO (4 mL), DIPEA (4 mL) was added at room temperature and the reaction mixture was stirred at 120 °C overnight. The reaction mixture was diluted with water, extracted with EtOAc, and the combined organic phase was washed with water (20 mL), brine (20 mL), dried over anhydrous sodium sulfate, and concentrated to give a residue. The residue was purified by flash chromatography (elution gradient: 0% to 30% EtOAc in PE, 30 min) to give methyl 5-(3-(tert-butoxycarbonylamino)-3-(methylcarbamoyl)piperidin-1-yl)-2-(3-fluorophenyl)isonicotinate (78-6). LC-MS: [M+H] + =487.3.

[0421] To a mixture of methyl 5-(3-(tert-butoxycarbonylamino)-3-(methylcarbamoyl)piperidin-1-yl)-2-(3-fluorophenyl)isonicotinate (78-6) (150 mg, 0.308 mmol), LiCl (130 mg, 3.08 mmol), THF (5 mL), and MeOH (5 mL) was added NaBH 4 (232 mg, 6.16 mmol) was added portionwise. After the addition, the mixture was stirred at room temperature for 3 h. The reaction was then quenched by adding water (10 mL), extracted with EtOAc, and the combined organic phase was washed with water (20 mL), brine, dried over anhydrous sodium sulfate, and concentrated to give a residue. The residue was purified by flash chromatography (elution gradient: 10% to 50% EtOAc in PE, 30 min) to give tert-butyl 1-(6-(3-fluorophenyl)-4-(hydroxymethyl)pyridin-3-yl)-3-(methylcarbamoyl)piperidin-3-ylcarbamate (78-7). LC-MS: [M+H] + =459.2.

[0422] To a solution of tert-butyl 1-(6-(3-fluorophenyl)-4-(hydroxymethyl)pyridin-3-yl)-3-(methylcarbamoyl)piperidin-3-ylcarbamate (78-7) (129 mg, 0.282 mmol), intermediate B (142 mg, 0.423 mmol), and n-Bu3P (171 mg, 0.845 mmol) in THF (5 mL) was added DIAD (171 mg, 0.845 mmol) 2 After the addition, the mixture was subjected to N 2 The mixture was stirred under reduced pressure for 1 h at room temperature, quenched with water and extracted with EtOAc. The combined organic phase was worked up under aqueous conditions and concentrated to give a residue. The residue was purified by flash chromatography (elution gradient: 10% to 70% EtOAc in PE, 30 min) to give tert-butyl (tert-butoxycarbonyl) (9-(5-(3-(tert-butoxycarbonyl)amino)-3-(methylcarbamoyl)piperidin-1-yl)-2-(3-fluorophenyl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate (78-8). LC-MS: [M+H] + =776.3

[0423] To a solution of tert-butyl (tert-butoxycarbonyl) (9-((5-(3-((tert-butoxycarbonyl)amino)-3-(methylcarbamoyl)piperidin-1-yl)-2-(3-fluorophenyl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate (78-8) (100 mg, 0.232 mmol) in DCM (9 mL) was added TFA (3 mL) at room temperature. The reaction mixture was stirred at room temperature for 1 h. The mixture was concentrated in vacuo to give a residue. The residue was purified by preparative HPLC (basic conditions, NH 3 H 2 O% = 0.05%, MEOH / H 2O = 0-95%, 12 min) to give 40 mg of pure racemic product, which was isolated by SFC to give (R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(3-fluorophenyl)pyridin-3-yl)-N-methylpiperidine-3-carboxamide (Example 78). 1 H NMR (400MHz, CD 3 OD)δ 8.39(s,1H),8.17(s,1H),8.11(s,1H),7.44-7.39(m,2H),7.32-7.27(m,1H),7.18(s,1H),7.01-6.97(m,1H),5.62-5.51(m,2H),3.32-3. 29(m,1H),3.02-2.99(m,1H),2.91-2.78(m,2H),2.69(s,3H),2.09-2 .01(m,1H),1.97-1.87(m,1H),1.70-1.67(m,1H),1.55-1.51(m,1H). LC-MS:[M+H] + =476.2

[0424] Following procedures similar to those described in Example 78, Examples 79-88 can be prepared. [Table 7-1] [Table 7-2] [Table 7-3]

[0425] Example 89: 1-(3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(3,4-difluorophenyl)pyridin-3-yl)piperidin-3-yl)-2,3-dimethylbutan-1-one [ka] To a solution of methyl (3-(2,3-dimethylbutanoyl)piperidin-3-yl)carbamate (Intermediate 89-6) (367 mg, 1.4 mmol, 1.0 equiv.) and methyl 2-(3,4-difluorophenyl)-5-fluoroisonicotinate (Intermediate A) (383 mg, 1.4 mmol, 1.0 equiv.) in DMSO (10 mL) was added DIPEA (724 mg, 5.6 mmol, 4.0 equiv.). The mixture was diluted with N 2 The mixture was stirred at 120° C. for 5 hours under atmospheric pressure. 2 The mixture was diluted with 20 mL of 2H2O and extracted with EtOAc (20 mL x 2). The combined organic layers were washed with brine (30 mL) and anhydrous Na 2 SO 4 The residue was purified by silica gel column chromatography (30% EtOAc in PE) to give methyl 2-(3,4-difluorophenyl)-5-(3-(2,3-dimethylbutanoyl)-3-((methoxycarbonyl)amino)piperidin-1-yl)isonicotinate (89-7). 1 H NMR (400MHz, CD 3 OD)δ ppm 8.52(s,0.5H),8.50(s,0.5H),8.02(s,0.5H),7.98(s,0.5H),7.94-7.86(m,1H),7.80-7.73(m,1H),7.3 9-7.30(m,1H),4.00(s,1.5H),3.99(s,1.5H),3.75(d,J=12.4Hz,0.5H),3.67(d,J=12.0Hz,0.5H),3.59 (s,1.5H),3.54(s,1.5H),3.42-3.34(m,1H),3.23(d,J=12.4Hz,0.5H),3.13-3.00(m,1.5H),2.97-2.88 (m,1H),2.30-2.20(m,1H),2.01-1.82(m,3H),1.78-1.66(m,1H),0.99-0.94(m,3H),0.90-0.84(m,6H). LC-MS:[M+H] + =504.2.

[0426] CH 3To a mixture of methyl 2-(3,4-difluorophenyl)-5-(3-(2,3-dimethylbutanoyl)-3-((methoxycarbonyl)amino)piperidin-1-yl)isonicotinate (89-7) (416 mg, 0.82 mmol, 1.0 equiv) in OH (10 mL) was added NaBH 4 (312 mg, 8.2 mmol, 10 equiv.) was added at room temperature. The mixture was stirred at room temperature for 20 min. Another NaBH 4 (312 mg, 8.2 mmol, 10 equiv) was added and the mixture was stirred at room temperature for an additional 20 min. 2 The mixture was diluted with 20 mL of 2H2O and extracted with EtOAc (20 mL x 2). The combined organic layers were washed with brine (20 mL) and anhydrous Na 2 SO 4 The residue was purified by silica gel column chromatography (30% EtOAc in hexane) to give methyl (1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridin-3-yl)-3-(1-hydroxy-2,3-dimethylbutyl)piperidin-3-yl)carbamate (89-8). 1 H NMR (400MHz, CD 3 OD)δ ppm 8.34(s,0.4H),8.33(s,0.6H),7.91(s,0.4H),7.90(s,0.6H),7.88-7.81(m,1H),7.76-7 .70(m,1H),7.39-7.32(m,1H),4.84-4.69(m,1H),3.90-3.64(m,1H),3.63-3.55(m,3H), 3.49-3.43(m,0.3H),3.28-3.23(m,0.6H),3.18-3.02(m,2H),2.86-2.76(m,1H),2.31-2 .15(m,1H),2.00-1.86(m,1H),1.82-1.54(m,3H),1.21-1.20(m,1H),0.95-0.78(m,9H). LC-MS:[M+H] + =478.2.

[0427] A mixture of methyl (1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridin-3-yl)-3-(1-hydroxy-2,3-dimethylbutyl)piperidin-3-yl)carbamate (Intermediate 89-8) (273 mg, 0.57 mmol, 1.0 equiv.), Intermediate B (192 mg, 0.57 mmol, 1.0 equiv.), and Bu in THF (15 mL). 3 To a solution of P (578 mg, 2.86 mmol, 5.0 equiv.), DIEA (578 mg, 2.86 mmol, 5.0 equiv.) was added 2 The reaction mixture was then cooled to 0° C. 2 The mixture was stirred at room temperature under H 2 The mixture was diluted with 20 mL of 2H2O and extracted with EtOAc (20 mL x 2). The combined organic layers were washed with brine (30 mL) and anhydrous Na 2 SO 4 The residue was purified by silica gel column chromatography (50% EtOAc in PE) to give tert-butyl (tert-butoxycarbonyl) (9-((2-(3,4-difluorophenyl)-5-(3-(1-hydroxy-2,3-dimethylbutyl)-3-((methoxycarbonyl)amino)piperidin-1-yl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate (89-9). 1 H NMR (400MHz, CD 3 OD)δ ppm 8.91-8.81(m,1H),8.80-8.70(m,1H),8.56-8.50(m,1H),7.76-7.67(m,1H),7.59-7.51(m,1H),7.36-7.23(m,2H),5.92-5.64(m,2H) ),4.04-3.65(m,2H),3.60-3.48(m,3H),3.30-2.75(m,3H),2.33-2.13(m,2H),1.88-1.70(m,4H),1.36(s,18H),0.98-0.93(m,9H). LC-MS:[M+H] + =795.3.

[0428] A mixture of tert-butyl (tert-butoxycarbonyl) (9-((2-(3,4-difluorophenyl)-5-(3-(1-hydroxy-2,3-dimethylbutyl)-3-(methoxycarbonyl)amino)piperidin-1-yl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate (89-9) (279 mg, 0.37 mmol, 1.0 equiv.) and DMP (237 mg, 0.56 mmol, 1.5 equiv.) in DCM (10 ml) was stirred at room temperature for 4 h. The mixture was diluted with saturated NaHCO 3 The mixture was quenched with aqueous solution (30 mL) and extracted with DCM (30 mL x 2). The combined organic layers were washed with saturated NaHCO 3 Wash with aqueous solution (20 mL x 2) and saline (30 mL), and then with anhydrous Na 2 SO 4 The residue was purified by silica gel column chromatography (50% EtOAc in PE) to give tert-butyl (tert-butoxycarbonyl) (9-((2-(3,4-difluorophenyl)-5-(3-(2,3-dimethylbutanoyl)-3-((methoxycarbonyl)amino)piperidin-1-yl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate (89-10). LC-MS: [M+H] + =793.5.

[0429] A mixture of tert-butyl (tert-butoxycarbonyl)(9-((2-(3,4-difluorophenyl)-5-(3-(2,3-dimethylbutanoyl)-3-((methoxycarbonyl)amino)piperidin-1-yl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate (89-10) (280 mg, crude, 0.34 mmol, 1 equiv.) in acetic acid and 10 mL of 35% HBr was stirred at room temperature for 20 h. LC-MS showed that most of the starting material had been consumed. The reaction mixture was concentrated in vacuo and H 2 The aqueous layer was diluted with 20 mL of EtOAc (30 mL×3) and washed with EtOAc (30 mL×3) to remove impurities. 3 H 2The mixture was basified to pH = 11 with O and extracted with EtOAc (30 mL x 2). 2 SO 4 Drying at 40° C., filtration and concentration gave the crude product which was purified by preparative HPLC to give 1-(3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(3,4-difluorophenyl)pyridin-3-yl)piperidin-3-yl)-2,3-dimethylbutan-1-one (Example 89). 1 H NMR (400MHz, CD 3 OD)δ 8.52(d,J=4.0Hz,1H),8.32-8.17(m,2H),7.83-7.65(m,1H),7.60-7.48(m,1H),7.37-7.15(m,2H),5.67(d,J=3.6Hz,2H),3.4 1(t,J=12.0Hz,1H),3.14-2.87(m,4H),2.08-1.61(m,5H),1.06(t,J=8.0Hz,3H),0.95(d,J=6.8Hz,3H),0.90(d,J=6.8Hz,3H). LC-MS:[M+H] + =535.3.

[0430] Following procedures similar to those described in Example 89, Examples 90-112 can be prepared. [Table 8-1] [Table 8-2] [Table 8-3] [Table 8-4] [Table 8-5] [Table 8-6] [Table 8-7] [Table 8-8]

[0431] Example 113 and Example 114: (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(3,4-difluorophenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol and (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(3,4-difluorophenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol [ka] To a solution of tert-butyl (3-(2,2-difluoro-1-hydroxyethyl)piperidin-3-yl)carbamate (Intermediate 113-5) (1.2 g, 4.28 mmol) and methyl 2-(3,4-difluorophenyl)-5-fluoroisonicotinate (Intermediate A) (1.144 g, 4.28 mmol) in DMSO (30 mL), DIPEA (14.95 mL, 86 mmol) was added at room temperature and the reaction mixture was cooled to 5° C. with N 2 The mixture was stirred under atmospheric pressure at 120° C. for 8 hours. The reaction mixture was diluted with water (40 mL) and extracted with EtOAc (30 mL×3). The combined organic layer was washed with water (20 mL×3), brine (20 mL), and anhydrous Na 2 SO 4 The mixture was dried over 1000 ml of ethyl acetate, filtered and concentrated to give a residue. The residue was purified by flash chromatography (elution gradient: 10% to 50% EtOAc in PE, 40 min) to give methyl 5-(3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidin-1-yl)-2-(3,4-difluorophenyl)isonicotinate (113-6). LC-MS: [M+H] + =528.2.

[0432] To a solution of methyl 5-(3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidin-1-yl)-2-(3,4-difluorophenyl)isonicotinate (113-6) (1.2 g, 2.275 mmol) in THF (30 mL) and MeOH (30 mL) was added lithium chloride (1.929 g, 45.5 mmol) and sodium tetrahydroborate (1.721 g, 45.5 mmol) at 0° C., and the reaction mixture was irradiated with N 2 The mixture was stirred at room temperature under atmosphere for 4 hours. The reaction mixture was diluted with 30 mL of water and extracted with EtOAc (20 mL×3). The combined organic layers were washed with water (20 mL), brine (20 mL) and concentrated to give the crude product. The crude product was purified by flash chromatography (elution gradient: 20% to 70% EtOAc in PE, 40 min) to give tert-butyl (3-(2,2-difluoro-1-hydroxyethyl)-1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridin-3-yl)piperidin-3-yl)carbamate (113-7). LC-MS: [M+H] + =500.2.

[0433] To a solution of tert-butyl (3-(2,2-difluoro-1-hydroxyethyl)-1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridin-3-yl)piperidin-3-yl)carbamate (Intermediate 113-7) (800 mg, 1.602 mmol), Intermediate B (645 mg, 1.922 mmol), and triphenylphosphine (1260 mg, 4.80 mmol) in THF (20 mL) was added DEAD (0.761 mL, 4.80 mmol) dropwise at 0 °C. After addition, the reaction mixture was immersed in N 2The mixture was stirred at 0° C. under atmosphere for 0.5 h. The reaction mixture was diluted with 20 mL of water and extracted with EtOAc (20 mL×3). The combined organic layers were washed with water (20 mL), brine (20 mL), and concentrated to give the crude product. The crude product was purified by flash chromatography (elution gradient: 20% to 70% EtOAc in PE, 50 min) to give tert-butyl (tert-butoxycarbonyl) (9-(5-(3-(tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidin-1-yl)-2-(3,4-difluorophenyl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate (113-8). LC-MS: [M+H] + =817.2.

[0434] To a solution of tert-butyl (tert-butoxycarbonyl)(9-((5-(3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidin-1-yl)-2-(3,4-difluorophenyl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate (113-8) (600 mg, 0.735 mmol) in DCM (30 mL) was added TFA (10 mL) and the reaction mixture was cooled to 37° C. with N 2 The mixture was stirred at room temperature under atmospheric pressure for 2 hours. The reaction mixture was concentrated in vacuo to give the crude product. The crude product was purified by preparative HPLC (basic condition NH3H2O%=0.05%, MEOH / H2O=0-95%, 10 min) to give 330 mg of pure racemic product. The racemic products were isolated by SFC to give (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(3,4-difluorophenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol (Example 113) and (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(3,4-difluorophenyl)pyridin-3-yl)-2,2-difluoroethan-1-ol (Example 114).

[0435] Example 113: 1H NMR (CD 3 OD,400MHz)δ(ppm):8.48(s,1H),8.22(d,2H),7.71-7.77(m,1H),7.51-7.54(m,1H),7.29(s,1H),7.24-7.27(t,1H),5.9 1-6.19(t,1H),5.65(s,2H),3.65(br,1H),3.20(d,1H),2.99(m,2H),2.84(t,1H),1.93-1.99(m,1H),1.66~1.86(m,3H). LC-MS:[M+H] + =517.2.

[0436] Example 114: 1 H NMR (CD 3 OD-d6,400MHz)δ(ppm):8.49(s,1H),8.22(d,2H),7.69-7.75(m,1H),7.48-7.51(m,1H),7.24-7.31(t,1H),7.21(s, 1H), 5.89-6.18(t,1H),5.65(s,2H),3.95(br,1H),3.25(d,1H),2.90-3.10(b,3H),1.82-1.88(m,3H),1.63(br,1H). LC-MS:[M+H] + =517.2.

[0437] Following procedures similar to those described in Examples 113 and 114, Examples 115-155 can be prepared. [Table 9-1] [Table 9-2] [Table 9-3] [Table 9-4] [Table 9-5] [Table 9-6] [Table 9-7] [Table 9-8] [Table 9-9] [Table 9-10] [Table 9-11] [Table 9-12] [Table 9-13]

[0438] Example 156 and Example 157: (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(2,4,5-trifluorophenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol and (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(2,4,5-trifluorophenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol [ka] Step 1. 1-(3-amino-1-(4-(hydroxymethyl)-6-(2,4,5-trifluorophenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol (Intermediate 156-2) (700 mg, 1.677 mmol, 1.0 equiv.), Intermediate B (80 mg, 0.240 mmol, 1.5 equiv.), and n-Bu in anhydrous THF (7 mL). 3To a solution of P (509 mg, 2.516 mmol, 1.5 equiv) was added DIAD (509 mg, 2.516 mmol, 1.5 equiv) under nitrogen at 0° C. The reaction mixture was stirred at 0° C. for 30 min. The reaction mixture was concentrated in vacuo and the residue was purified by preparative HPLC (base) to give intermediate 156-3. 1 H NMR (400 MHz, CDCl 3 )δ=8.93- 8.85(m,1H),8.57(s,1H),8.22-8.14(m,1H),7.92-7.76(m,1H),7.45-7.32(m,1H),7.01-6.83(m,1H),6.20-5.80(m,1H),5.72-5.53(m,2) H),3.82-3.63(m,1H),3.26-3.06(m,2H),3.05-2.89(m,2H),2.28-2.12(m,1H),1.89-1.79(m,2H),1.75-1.63(m,1H),1.49-1.36(m,18H). LCMS:[M+H] + =735.5.

[0439] Step 2. To a solution of intermediate 156-3 (630 mg, 0.858 mmol) in anhydrous DCM (15 mL) was added TFA (5 mL) and the reaction was stirred for 3 h at 20° C. The reaction mixture was concentrated in vacuo, basified with ammonium hydroxide to pH=8, and concentrated in vacuo. The residue was purified by preparative HPLC (base) and SFC to give (R)-1-((R)-3-amino-1-(4-(6-amino-9H-purin-9-yl)methyl)-6-(2,4,5-trifluorophenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol (Example 156) and (S)-1-((R)-3-amino-1-(4-(6-amino-9H-purin-9-yl)methyl)-6-(2,4,5-trifluorophenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol (Example 157).

[0440] Example 156: 1H NMR (400MHz, CD3OD) δ ppm 8.52(d,J=1.9Hz,1H),8.20(dd,J=5.6,2.0Hz,2H),7.78(dddd,J=11.4,9.2, 7.1,2.0Hz,1H),7.42-7.05(m,2H),6.35-5.80(m,1H),5.67(d,J=2.0Hz,2H) ,3.68(d,J=14.0Hz,1H),3.22(s,1H),3.14-3.00(m,2H),2.88(t,J=11.2Hz, 1H),2.03(t,J=11.5Hz,1H),1.82(t,J=12.9Hz,2H),1.71(d,J=13.0Hz,1H). LC-MS: [M+H]+=535.2.

[0441] Example 157: 1 H NMR(400MHz,CD3OD)δ ppm 8.55(s,1H),8.25(s,1H),8.21(s,1H),7.87-7.74(m,1H),7.25-7.15(m,1H),7.09(s,1H),6.26-5.88(m,1H) ,5.70(s,2H),4.10-3.90(m,1H),3.30-3.20(m,1H),3.17-2.87(m,3H),2.03-1.80(m,3H),1.75-1.60(m,1H). LC-MS:[M+H]+=535.4.

[0442] Following a procedure similar to the preparation of Examples 156 and 157 (Step 2), Examples 158 and 159 were prepared. [Table 10]

[0443] Example 160 and Example 161: (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(2,5-difluoro-4-methoxyphenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol and (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(2,5-difluoro-4-methoxyphenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol [ka] To a solution of tert-butyl (tert-butoxycarbonyl)(9-((5-(3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidin-1-yl)-2-(2,5-difluoro-4-methoxyphenyl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate (Intermediate 160-3) (200 mg, 0.237 mmol) in DCM (18 mL) was added TFA (36 mL) and the reaction mixture was cooled to 4° C. with N 2 Stirred at room temperature under atmosphere for 30 minutes. The reaction mixture was concentrated in vacuum to obtain crude product. The crude product was purified by preparative HPLC and SFC to obtain (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(2,5-difluoro-4-methoxyphenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol (Example 160) and (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(2,5-difluoro-4-methoxyphenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol (Example 161).

[0444] Example 160: 1 H NMR (400MHz, CD 3OD)δ ppm 8.48(s,1H),8.20(d,J=1.6Hz,2H),7.58(dd,J=12.2,7.3Hz,1H),7.11(d,J=1.3Hz,1H),6.90(dd,J=12.6,7.1Hz,1H ),6.06(td,J=55.1,3.9Hz,1H),5.67(s,2H),3.87(s,3H),3.75-3.58(m,1H),3.25-2.75(m,4H),2.26-1.60(m,4H). LC-MS:[M+H] + =547.2,548.2.

[0445] Example 161: 1 H NMR (400MHz, CD 3 OD)δ=8.51-8.44(m,1H),8.24-8.16(m,2H),7.62-7.48(m,1H),7.03(s,1H),6.93-6.79(m,1H),6.25- 5.86(m,1H),5.71-5.59(m,2H),4.00(m,1H),3.88-3.80(m,3H),3.28-2.87(m,4H),1.99-1.56(m,4H). LC-MS:[M+H] + =547.4.

[0446] Example 162 and Example 163: (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(2-(difluoromethyl)-4-methoxyphenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol and (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(2-(difluoromethyl)-4-methoxyphenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol [ka] Intermediate 162-2 (57 mg, 0.129 mmol), Intermediate B (43.1 mg, 0.129 mmol), and PPh in THF (5 mL) 3(101 mg, 0.386 mmol) was added DEAD (0.061 mL, 0.386 mmol). The reaction mixture was cooled to 5° C. 2 The mixture was stirred at 0° C. under ambient atmosphere for 30 min. The reaction mixture was quenched with water, extracted with EtOAc, and the combined organic phase was washed with water, brine, dried over sodium sulfate, and concentrated to give the crude product. The crude product was purified by flash chromatography (elution gradient: 0% to 10% MeOH in DCM for 30 min) to give the intermediate. LC-MS: [M+H] + =761.

[0447] To a solution of the intermediate (200 mg, 0.263 mmol) in DCM (18 mL) was added TFA (36 mL, 467 mmol) and the reaction mixture was rinsed with N 2 The mixture was stirred at room temperature under atmospheric pressure for 30 minutes. The crude product was purified by preparative HPLC (basic condition, NH 3 H 2 O% = 0.05%, MeOH / H 2 O = 0 ~ 95%, 10 min) to give the pure product, which was then further purified to give (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl) methyl)-6-(2-(difluoromethyl)-4-methoxyphenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol (Example 162) and (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purin-9-yl) methyl)-6-(2-(difluoromethyl)-4-methoxyphenyl)pyridin-3-yl)piperidin-3-yl)-2,2-difluoroethan-1-ol (Example 163).

[0448] Example 162: 1 H NMR (400MHz, CD 3OD)δ ppm 8.48(s,1H),8.21(d,J=3.7Hz,2H),7.34-7.26(m,1H),7.17(d,J=2.6Hz,1H),7.07(dt ,J=8.6,1.8Hz,1H),6.95-6.61(m,2H),6.07(td,J=55.1,3.9Hz,1H),5.68(s,2H),3.76 -3.61(m,1H),3.22(d,J=11.4Hz,1H),3.17-3.08(m,1H),3.04(d,J=11.4Hz,1H),2.93 -2.81(m,1H),2.02(dd,J=13.9,8.9Hz,1H),1.91-1.75(m,2H),1.70(d,J=12.7Hz,1H). LC-MS:[M+H]+=560.9,562.0.

[0449] Example 163: 1 H NMR (400MHz, CD 3 OD)δ ppm 8.55(s,1H),8.30(d,J=14.6Hz,2H),7.34(d,J=8.6Hz,1H),7.20(d,J=2.6Hz,1H),7.10(dd,J=8.6,2.6Hz,1H),7.01(s,1H),6.85(t,J=55 .3Hz,1H),6.19(td,J=53.9,2.6Hz,1H),5.68(d,J=3.3Hz,2H),4.20(s,1H),3.50-3.34(m,2H),3.19-2.98(m,2H),2.02(d,J=42.5Hz,4H). LC-MS:[M+H]+=560.9,562.0.

[0450] Following similar procedures to the preparation of Examples 162 and 163, Examples 164-167 were prepared from Intermediate B and the corresponding intermediates. [Table 11-1] [Table 11-2] [Table 11-3]

[0451] Following the same procedures as in the preparation of Example 168 and Example 169, Examples 168-176 were prepared from the corresponding intermediates. [Table 12-1] [Table 12-2] [Table 12-3] [Table 12-4]

[0452] Example 177: 9-((5-(3-amino-3-(6-methylpyridin-2-yl)piperidin-1-yl)-2-(3,4-difluorophenyl)pyridin-4-yl)methyl)-9H-purin-6-amine [ka] A solution of tert-butyl (tert-butoxycarbonyl)(9-((2-(3,4-difluorophenyl)-5-(3-((methoxycarbonyl)amino)-3-(6-methylpyridin-2-yl)piperidin-1-yl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate (177-7) (700 mg, 0.89 mmol, 1.0 equiv) in HBr / AcOH (33%, 10 mL) was stirred at 25-30 °C for 16 h. The reaction was concentrated in vacuo. The residue was purified by preparative HPLC (0.1% NH as additive). 4 HCO 3 ) and purified by 9-((5-( 3 -amino-3-(6-methylpyridin-2-yl)piperidin-1-yl)-2-(3,4-difluorophenyl)pyridin-4-yl)methyl)-9H-purin-6-amine (Example 177). 1 H NMR (400MHz, DMSO-d 6)δ ppm 8.53(s,1H),8.29(s,1H),8.14(s,1H),7.93-7.82(m,1H),7.72-7.65(m,1H),7.64-7.59(m,1 H),7.56(d,J=7.6Hz,1H),7.52-7.42(m,1H),7.37(s,1H),7.29(brs,2H),7.12(d,J=7.6Hz,1H ),5.55(dd,J=16.0Hz,28.4Hz,2H),3.44(d,J=11.2Hz,1H),3.17-3.08(m,1H),3.06-3.01(m, 1H), 3.00-2.90 (m, 1H), 2.47 (s, 3H), 2.24-2.14 (m, 1H), 2.13-1.99 (m, 1H), 1.77-1.65 (m, 1H). LC-MS:[M+H] + =528.3.

[0453] Following similar procedures as in the preparation of Example 177 and the corresponding intermediates, Examples 178-284 were prepared. [Table 13-1] [Table 13-2] [Table 13-3] [Table 13-4] [Table 13-5] [Table 13-6] [Table 13-7] [Table 13-8] [Table 13-9] [Table 13-10]

Table 13-11

Table 13-12

Table 13-13

Table 13-14

Table 13-15

Table 13-16

Table 13-17

Table 13-18

Table 13-19

Table 13-20

Table 13-21

Table 13-22

Table 13-23

Table 13-24

Table 13-25

Table 13-26

[0454] Examples 285-291 were prepared from the corresponding intermediates following procedures similar to those described in Example 47. [Table 14-1] [Table 14-2] [Table 14-3] [Table 14-4] [Table 14-5] [Table 14-6]

[0455] Example 301: 3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(3,4-difluorophenyl)pyridin-3-yl)-3-(2,2-difluoroethyl)piperidin-4-ol [ka] To a solution of intermediate B (104 mg, 0.311 mmol) and intermediate 301-8 (130 mg, 0.311 mmol) in DMF (10 mL) was added K 2 CO 3 (215 mg, 1.556 mmol) 2 The mixture was added at room temperature under atmospheric pressure and the reaction was stirred at room temperature for 18 hours. The reaction mixture was diluted with water and extracted with EtOAc (20 mL x 3), and the combined organic phase was washed with water (20 mL x 3), brine (20 mL), dried over anhydrous sodium sulfate, and concentrated in vacuo to give a residue. The residue was purified by flash chromatography (elution gradient: 0% to 10% MeOH in DCM, 30 min) to give intermediate 301-9. LC-MS: [M+H] + =311.1.

[0456] To a solution of intermediate 301-9 (60 mg, 0.084 mmol) in DCM (9 mL), TFA (3 mL, 38.9 mmol) was added N 2 The mixture was added at room temperature under atmospheric pressure and the reaction was stirred at room temperature for 2 hours. The reaction mixture was concentrated in vacuo to give a residue. The residue was purified by preparative HPLC (basic conditions, NH 3 H 2 O% = 0.05%, MeCN / H2 O=0-95%, 12 min) to give 3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(3,4-difluorophenyl)pyridin-3-yl)-3-(2,2-difluoroethyl)piperidin-4-ol (Example 301). 1 H NMR (400MHz, CD 3 OD)δ 8.58(s,1H),8.36(s,1H),8.27(s,1H),7.82(ddd,J=11.2,8.1,2.2Hz,1H),7.63( dt,J=5.8,2.9Hz,1H),7.47(s,1H),7.31(dt,J=10.2,8.4Hz,1H),6.41(tt,J=55. 3,4.7Hz,1H),5.65(s,2H),3.94(dd,J=10.4,5.3Hz,1H),3.50-3.38(m,2H),3.11 -2.93(m,2H),2.69-2.36(m,2H),2.07(dd,J=14.1,4.8Hz,1H),2.01-1.84(m,1H). LC-MS:[M+H]+=517.2.

[0457] Following procedures similar to those used to prepare Example 47 and the corresponding intermediates, Examples 302-310 were prepared. [Table 15-1] [Table 15-2]

[0458] Example 306: 9-((5-((3R,5R)-3-amino-5-(fluoromethyl)piperidin-1-yl)-2-(3-fluoro-4-methoxyphenyl)pyridin-4-yl)methyl)-9H-purin-6-amine [ka] To a mixture of tert-butyl (tert-butoxycarbonyl) (9-((5-((3R,5R)-3-((tert-butoxycarbonyl)amino)-5-(fluoromethyl)piperidin-1-yl)-2-(3-fluoro-4-15 methoxyphenyl)pyridin-4-yl)methyl)-9H-purin-6-yl)carbamate (Intermediate 306-9) (0.15 g, 0.19 mmol) in DCM (1 mL) was added HCl / dioxane (10.0 mL, 4 M) and the resulting mixture was stirred at 25° C. for 4 h. The mixture was concentrated in vacuo. The residue was purified by preparative HPLC (0.1% NH as additive). 4 HCO 3 ) to give 9-((5-((3R,5R)-3-amino-5-(fluoromethyl)piperidin-1-yl)-2-(3-fluoro-4-methoxyphenyl)pyridin-4-yl)methyl)-9H-purin-6-amine (Example 306). 1 H NMR (400MHz, DMSO-d 6 ):δ 8.48(s,1H),8.33(s,1H),8.14(s,1H),7.67(dd,J=2.0Hz,12.8Hz,1H),7.51(d,J=8.8Hz,1H),7.30(brs,2H),7.34-7.17(m,2 H),5.54(s,2H),4.64-4.37(m,2H),3.85(s,3H),3.18-3.03(m,3H),2.92-2.77(m,2H),2.48-2.32(m,1H),1.68-1.49(m,2H). LC-MS:[M+Na] + =481.2

[0459] Following similar procedures as in the preparation of Example 306 and the corresponding intermediates, Examples 307-310 were prepared. [Table 16]

[0460] Following the same procedure as for the preparation of Example 47 and the corresponding intermediates, Examples 311-321 were prepared. [Table 17-1] [Table 17-2] [Table 17-3]

[0461] Example 322: (S)-3-(3-amino-1-(4-((6-amino-9H-purin-9-yl)methyl)-6-(3,4-difluorophenyl)pyridin-3-yl)piperidin-3-yl)-N,N-dimethylpropanamide [ka]

[0462] Following procedures similar to the preparation of Example 89 and the corresponding intermediates, Example 322 was prepared. 1 H NMR (400MHz, CD 3 OD)δ:8.50(s,1H),8.24(d,J=10.8Hz,2H),7.73(ddd,J=12.3,7.7,2.3Hz,1H),7.51(t,J=5.7Hz,1H),7.28(d,J=8.5Hz,2H),5. 85-5.51(m,2H),3.10(s,3H),2.99(d,J=19.4Hz,4H),2.93(s,3H),2.53(t,J=8.1Hz,2H),2.08-1.71(m,4H),1.71-1.41(m,2H). LC-MS:[M+H] + =535.8,536.8.

[0463] Following similar procedures to the preparation of Example 1 and the corresponding intermediates, Examples 323-325 were prepared. [Table 18]

[0464] Biological assays Compounds of the invention may be evaluated for their ability to inhibit NSD2 using the assays described below and others known in the art.

[0465] LC-MS / MS-based NSD2 enzyme assay In this assay, LC-MS / MS technology was used to monitor SAH production from the NSD2 enzymatic reaction. The enzymatic reaction was performed on white proxy plates and 384-well microplates (Perkin Elmer). The reaction mixture (10 μL) consisted of 8 nM NSD2[1-1365], 1 μM SAM (USB, 10601), and 400 nM nucleosomes (purified from mouse liver) in reaction buffer (20 mM Tris-HCl (pH 8.0), 0.01% Tween 20, 10 mM MgCl2, 50 mM NaCl, 1 mM DTT, 1 mM TCEP (pH 7.5)). After 90 min of incubation at room temperature, the reaction was stopped by adding 3 μL of quenching solution containing 320 nM d4-SAH in 2.5% TFA.

[0466] For inhibition assays, compound solutions were transferred to wells by Mosquito™ (TTP LabTech). Inhibition assays were performed by preincubating various concentrations of inhibitors with a 5uL reaction mixture containing 16 nM NSD2[1-1365] and 2uM SAM (USB, 10601) in reaction buffer. After 20 minutes of preincubation, 5uL of a solution containing 800nM nucleosomes (purified from mouse liver) was added in reaction buffer to start the reaction. After 90 minutes, the reaction was stopped by 3μL of quenching solution containing 320nM d4-SAH in 2.5% TFA.

[0467] SAH production from the enzymatic assay was monitored by LC-MS / MS on an API4000 triple quadrupole mass spectrometer using a TurboIon Spray (Applied Biosystem) coupled with a Prominence UFLC (Shimadzu). Liquid chromatography was performed on a Chromolith FastGradient HPLC column (RP-18e, 25-2 mm, Merck) at a flow rate of 0.8 mL / min. The column was directly connected to a TurboIon Electrospray operated in positive ion mode. Mobile phase A was 0.1% FA and 2% methanol in water, and mobile phase B was 0.1% FA in ACN. The injection volume was 3 μl and the autosampler was kept at 4° C. SAH and d4-SAH were monitored simultaneously. Data were acquired and processed by Analyst software.

[0468] To quantify the SAH formed, d4-SAH was added as an internal standard (IS). A series of SAH (Sigma, A9384) solutions at various concentra...

Claims

1. Compounds of formula (I) 【Chemistry 1】 or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof, wherein: A is N or CR ９ and R ９ is hydrogen or halo, L is a bond or C １－４ is alkylene, R １ is H, or R １ and R ２ together with NH form a 5-8 membered heterocyclyl containing 1-2 heteroatoms selected from N, O and S as ring members, said 5-8 membered heterocyclyl being unsubstituted or substituted by an oxo substituent; R ２ teeth, (i) hydrogen, -C １－６ Alkyl, -haloC １－６ Alkyl, -hydroxy C １－６ Alkylene, -hydroxyhaloC １－６ Alkylene, -C １－６ Alkoxy C １－６ Alkylene, -haloC １－６ Alkoxy C １－６ Alkylene, or -C ３－８ Cycloalkoxy (C １－６ alkyl), (ii) cyano, -cyanoC １－６ Alkylene, -C １－６ Alkylthio C １－６ Alkyl, -C ２－６ Alkenyl, -haloC ２－６ Alkenyl, -C ２－６ Alkynyl, -C １－４ Alkyl SOC １－４ Alkyl, -C １－４ AlkylSO ２ C １－４ Alkyl, -SO ２ R ８ , or -C(C １－４ alkyl)=N-O(C １－４ alkyl), (iii)-C １－４ Alkylcarbonyl, -(CR ａ R ｂ ) ｐ -C(=O)-OR １０ or -C(=O)-(CR ａ R ｂ ) ｑ R １１ - (wherein, R １１ is C ３－７ cycloalkyl, 5- to 6-membered heterocyclyl, or 5- to 6-membered heteroaryl, each of which is independently unsubstituted or １－６ Alkyl or C １－６ substituted with alkoxy), (iv) -(CR ａ R ｂ ) ｒ -C(=O)-NR １２ R １３ (In the formula, R １２ is hydrogen or C １－６ alkyl, R １３ is hydrogen, -C １－６ alkyl or a 5- to 6-membered heterocycle, or R １２ and R １３ taken together form a 5- to 6-membered heterocyclic ring, which is unsubstituted, or １－４ substituted with alkyl) (v) 5- to 6-membered heterocyclyl C ０－６ Alkyl or 5-6 membered heterocyclyl (haloC １－４ alkyl), wherein each said heterocyclyl group is unsubstituted or substituted by oxo; (vi) 5- to 9-membered heteroaryl C ０－６ Alkyl or 5-9 membered heteroaryl (haloC １－４ wherein each said heteroaryl group is unsubstituted or is selected from the group consisting of -C １－４ Alkyl, -haloC １－４ Alkyl, -hydroxy C １－４ Alkylene, -C １－４ Alkoxy, -haloC １－４ Alkoxy, halo, hydroxy, cyano, oxide, aminocarbonyl C ０－６ Alkyl, -C １－４ Alkylaminocarbonyl C ０－６ Alkyl, -diC １－４ Alkylaminocarbonyl C ０－６ Alkyl or -C ３－７ substituted by cycloalkyl; R ３ａ , R ３ｂ , R ４ａ , R ４ｂ , R ５ａ , R ５ｂ , R ６ａ , and R ６ｂ are independently hydrogen, halo, cyano, hydroxyl, -C １－６ Alkyl, -haloC １－６ Alkyl, -hydroxy C １－６ Alkylene, -C １－６ Alkoxy, -C １－６ Alkoxy C １－６ Alkylene, -haloC １－６ Alkoxy C １－６ Alkylene, -hydroxyhaloC １－６ Alkylene, aryl, -C(=O)-OR １４ , or - (CR ａ R ｂ ) ｓ -C(=O)-NR １５ R １６ or R ３ａ and R ３ｂ , R ４ａ and R ４ｂ , R ５ａ and R ５ｂ , or R ６ａ and R ６ｂ forms an oxo substituent, R ７ is H, -C １－４ Alkoxy, halo, C １－４ alkyl, or 3-8 membered heterocyclyl unsubstituted or substituted by halo; R ８ is C ３－８ Cycloalkyl (C ０－６ alkyl); 4-6 membered heterocyclyl C containing 1-3 heteroatoms selected from N, O, and S ０－６ alkyl; aryl or 5-9 membered heteroaryl containing 1-3 heteroatoms selected from N, O, and S; ０－６ alkyl, R ８ is unsubstituted or has 1 to 3 R １７ is replaced by R １７ is halo, hydroxy, cyano, -C １－６ Alkyl, -haloC １－６ Alkyl, -C １－６ Alkoxy, -haloC １－６ Alkoxy, -NR ａ C(=O)CR ｃ = C(R ｃ ) ２ , or -(CR ａ R ｂ ) ｔ -NR ａ -C(=O)-R １８ and R ａ , R ｂ , R ｃ , R １０ , R １４ , R １５ , and R １６ are independently hydrogen or -C １－４ is alkyl, R １８ is -C １－４ Alkyl, or -C １－４ is haloalkyl, A compound of formula (I) as defined above, or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof, wherein p, q, r, s, and t are independently 0 to 4.

2. 2. A compound of formula (I) according to claim 1, wherein A is N, or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof.

3. R ３ａ is hydrogen or halo, R ３ｂ is hydrogen, halo, -hydroxyl, -C １－６ alkoxy or cyano, or R ４ａ is hydrogen or halo, R ４ｂ is hydrogen, halo, -C １－６ Alkoxy C １－６ Alkylene, -C １－６ Alkyl or -HaroC １－６ is alkyl, or R ５ａ is hydrogen, R ５ｂ is hydrogen or -C １－６ is alkyl, or R ５ａ and R ５ｂ 3. A compound of formula (I) according to claim 1 or 2, or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof, wherein together form an oxo substituent.

4. R ６ａ is hydrogen or halo; R ６ｂ is hydrogen, -haloC １－６ Alkoxy C １－６ Alkylene, -hydroxy C １－６ Alkylene, -hydroxyhaloC １－６ Alkylene, carboxyl, phenyl, or -(CR ａ R ｂ ) ｔ -C(O)-NR １５ R １６ and R ａ , R ｂ , R １５ , and R １６ are independently hydrogen or -C １－４ is alkyl, t is between 0 and 1; or R ６ａ and R ６ｂ A compound of formula (I) according to any one of claims 1 to 3, or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof, wherein together form an oxo substituent.

5. R ７ But H, -C １－４ 5. A compound of formula (I) according to any one of claims 1 to 4, or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof, which is alkoxy, or halo.

6. R １ , R ３ａ , R ３ｂ , R ４ａ , R ４ｂ , R ５ａ , R ５ｂ , R ６ａ , R ６ｂ , and R ７ A compound of formula (I) according to any one of claims 1 to 5, or a pharma- ceutically acceptable salt thereof, wherein is hydrogen.

7. The compound of formula (I) according to claim 1, wherein the compound is a compound of formula (II): 【Chemistry 2】 or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof.

8. R ２ is hydrogen, -C １－６ Alkyl, -HaroC １－６ Alkyl, -hydroxy C １－６ Alkylene, -hydroxyhaloC １－６ Alkylene, -C １－６ Alkoxy C １－６ Alkylene, -HaroC １－６ Alkoxy C １－６ Alkylene, or -C ３－８ Cycloalkoxy (C １－６ 8. The compound of claim 1, wherein R is an alkyl group, or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof.

9. R ２ is hydrogen, 2,2-difluoroethyl, 2-methyl-propan-1-olyl, ethane-1-olyl, 2,2-difluoroethane-1-olyl, 2-fluoroethane-1-olyl, 2,2,2-trifluoroethane-1-olyl, difluoromethoxyl, or 2,2,2-trifluoroethoxyl, or an enantiomer, mixture of enantiomers, or a pharma-ceutically acceptable salt thereof.

10. R ２ but, 【Chemistry 3】 8. The compound of claim 1, wherein:

11. R ２ But, -C １－４ Alkyl carbonyl, - (CR ａ R ｂ ) ｐ -C(O)-OR １０ (In the formula, R １０ is hydrogen or -C １－４ alkyl), -C(O)-(CR ａ R ｂ ) ｑ -R １１ (In the formula, R １１ But, C ３－６ cycloalkyl, 5- to 6-membered heterocyclyl, or 5- to 6-membered heteroaryl, each of which is independently unsubstituted or -C １－６ Alkyl or -C １－６ substituted with alkoxy), or - (CR ａ R ｂ ) ｒ -C(O)-NR １２ R １３ (Where, R １２ is hydrogen or -C １－６ is alkyl, R １３ is hydrogen, -C １－６ alkyl, or a 5- to 6-membered heterocycle; or R １２ and R １３が together form a 5- to 6-membered heterocyclic ring, said 5- to 6-membered heterocyclic ring being unsubstituted or １－４ substituted with alkyl; R ａ , R ｂ , and R １１ are independently hydrogen or -C １－４ is alkyl, 8. The compound of any one of claims 1 to 7, wherein p, q and r are independently 0 to 2, or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof.

12. R ２ but, pyridin-2-yl, (pyridin-2-yl)methyl, or (pyridin-2-yl)ethyl, wherein the pyridin-2-yl is independently unsubstituted or contains one to two -C １－４ Alkyl, -C １－４ Haloalkyl, hydroxyl, -Hydroxy C １－４ Alkylene, cyano, -C １－４ Alkoxy, -C １－６ Alkoxy C １－６ Alkylene, -haloC １－４ Alkoxy, halo or -C ３－７ substituted with cycloalkyl), Thiazol-4-yl, thiazol-2-ylmethyl, oxazol-2-yl, oxazol-2-ylmethyl, pyridazin-3-yl, pyridazin-3-ylmethyl, pyrazin-2-yl, pyrazin-2-ylmethyl, pyrimidin-4-yl, pyrimidin-4-ylmethyl, each of which is independently unsubstituted or -C １－４ substituted with alkyl or halo); 1H-pyrazol-3-yl, (1H-pyrazol-3-yl)methyl, 1-methyl-1H-pyrazol-4-yl, (1,2,3-triazol-4-yl), isoxazol-3-yl, each of which is independently unsubstituted or -C １－４ Alkyl, halo or -C ３－７ substituted with cycloalkyl), 1,4-dioxan-2-yl, pyridin-2(1H)-onyl, and 8. The compound according to any one of claims 1 to 7, or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof, selected from pyridin-1-oxid-2-yl.

13. R ８ 13. The compound of any one of claims 1 to 12, or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof, wherein is cyclopropyl, cyclobutyl, cyclohexyl, or azetidinyl.

14. R ８ is 2,3-dihydrobenzofuran-5-yl, 1,3-dihydroisobenzofuran-5-yl, benzo[d][1,3]dioxol-5-yl, imidazo[1,2-a]pyridin-6-yl, or pyrazolo[1,5-a]pyridin-6-yl, or an enantiomer, mixture of enantiomers, or a pharma- ceutically acceptable salt thereof.

15. R ８ is unsubstituted or has 1 to 2 halo, -C １－６ Alkyl or -C １－６ alkoxy substituted pyridyl; unsubstituted or substituted with 1-2 halo or -C １－６ 1H-indazol-5-yl, substituted with alkyl; or unsubstituted or -C １－６ 13. The compound of any one of claims 1 to 12, which is 1H-benzo[d]-imidazol-6-yl substituted with alkyl, or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof.

16. R ８ But 1 to 3 R １７ is a phenyl substituted with R １７ is halo, hydroxy, cyano, -C １－６ Alkyl, -haloC １－６ Alkyl, -C １－６ Alkoxy, -haloC １－６ Alkoxy, -NR ｄ C(O)CR ｄ = C(R ｄ ) ２ , or -(CR ａ R ｂ ) ｔ -NR ｄ -C(O)-R １８ and R ａ , R ｂ , and R ｄ are independently hydrogen or -C １－４ is alkyl, R １８ But, C １－４ is haloalkyl, 13. The compound according to any one of claims 1 to 12, or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof, wherein t is 0 to 1.

17. R ８ But 1 to 3 R １７ phenyl substituted with R １７ halo, hydroxy, -C １－６ Alkyl, -haloC １－６ Alkyl, -C １－６ Alkoxy, or -haloC １－６ 13. The compound of any one of claims 1 to 12, or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof, which is alkoxy.

18. 2. The compound of claim 1, or an enantiomer, a mixture of enantiomers, or a pharma- ceutically acceptable salt thereof, wherein the compound is selected from the compounds of Table 2 or a pharma- ceutically acceptable salt thereof.

19. 19. A pharmaceutical composition comprising a compound according to any one of claims 1 to 18, or a pharma- ceutically acceptable salt thereof, and one or more pharma- ceutically acceptable carriers.

20. A combination comprising a compound according to any one of claims 1 to 18, or a pharma- ceutically acceptable salt thereof, and one or more therapeutically active agents.

21. 21. The combination of claim 20, wherein the one or more therapeutically active agents are anti-cancer agents, analgesics, or anti-inflammatory agents.

22. 20. A method of treating a disease or condition mediated by nuclear SET domain-containing protein 2 (NSD2), comprising administering to a subject in need of such treatment a therapeutically effective amount of a compound according to any one of claims 1 to 18, or a pharma- ceutically acceptable salt thereof.

23. 23. The method of claim 22, wherein the disease or condition mediated by NSD2 is selected from solid tumors, leukemia, myeloma, lymphoma, and hypertension.

24. 24. The method of claim 23, wherein the disease or condition mediated by NSD2 is breast cancer, cervical cancer, skin cancer, ovarian cancer, gastric cancer, prostate cancer, pancreatic cancer, lung cancer, hepatocellular carcinoma, head and neck cancer, peripheral nerve sheath tumor, osteosarcoma, multiple myeloma, neuroblastoma, leukemia, non-Hodgkin's lymphoma, or pulmonary arterial hypertension.

25. 23. The method of claim 22, wherein the disease or condition mediated by NSD2 is acute lymphoblastic leukemia, cutaneous squamous cell carcinoma, or mantle cell lymphoma.

26. 20. Use of a compound according to any one of claims 1 to 18, optionally in combination with a second therapeutic agent, in the manufacture of a medicament for a disease or condition mediated by nuclear SET domain-containing protein 2 (NSD2).

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