Piperidinyl-methylpurine as an NSD2 inhibitor and anticancer agent

Abstract

This invention provides compounds, compositions, and methods for inhibiting nuclear SET domain-containing protein 2 (NSD2). [Solution] The present invention provides a compound of formula (I), or an enantiomer thereof, an enantiomer mixture thereof, or a pharmaceutically acceptable salt thereof. The present invention further provides pharmaceutical compositions comprising such compounds, and methods of using such compounds to treat diseases or conditions mediated by NSD2. TIFF2026143534000288.tif60156

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims the rights to PCT / CN2019 / 100542, filed on 14 August 2019, which is incorporated herein 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 Wolff-Hirschhorn syndrome candidate 1 (WHSC1) or multiple myeloma SET domain (MMSET), is an epigenetic modifier thought to play a driving role in oncogenesis. Both NSD2 overexpression and point mutations that increase its catalytic activity have been 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 NSD2 expression in T(4;14)+ MM cell lines is associated with increased H3K36me2 levels and an inversely proportional decrease in H3K27me3 levels. Such correlations suggest a causal relationship between NSD2 histone methyltransferase (HMT) activity and the MM oncogene.

[0005] High expression of the NSD2 protein has been shown in various human cancer species, including bladder cancer, brain cancer, gastrointestinal cancer, lung cancer, liver cancer, ovarian cancer, skin cancer, uterine cancer, breast cancer, prostate cancer, and glioblastoma (Coussens et al., Ezponda et al., Oncogene, 32:2882-2890 (2013)). In particular, NSD2 is one of the most frequently mutated genes in childhood cancer genomes. The NSD2SET domain variant E1099K, accompanied by an increase in H3K36me2 lacking the t(4;14) translocation, was identified in both acute lymphoblastic leukemia tumors and cell lines. Sequencing results from >1000 childhood 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, with both E1099K and T1150A variants observed. The E1099K variant has also been reported in chronic lymphocytic leukemia (CLL), lung cancer, and gastric cancer (Coussens et al., see above). In general, upregulation of NSD2 is associated with aggressive tumor behavior and poor prognosis (Ezponda et al., see above).

[0006] NSD2 is a promising target for cancer treatment, and there remains a need for selective inhibitors of NSD2. [Overview of the project]

[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 embodiment, the present invention relates to a compound of formula (I), [ka] or provides the enantiomer, enantiomer mixture, or pharmaceutically acceptable salt thereof, in the formula, A is N or CR 9 , and R 9 is hydrogen or halo, L is a bond or C 1-4 alkylene, R 1 is H, or R 1 and R 2 together with NH form a 5- to 8-membered heterocyclyl containing 1 to 2 heteroatoms selected from N, O and S as ring members, and said 5- to 8-membered heterocyclyl is unsubstituted or substituted by an oxo substituent, R 2 is (i) hydrogen, -C 1-6 alkyl, haloC 1-6 alkyl, hydroxyC 1-6 alkylene, hydroxyhaloC 1-6 alkylene, -C 1-6 alkoxyC 1-6 alkylene, haloC 1-6 alkoxyC 1-6 alkylene, or -C 3-8 cycloalkoxy(C 1-6 alkyl), (ii) cyano, cyano-C 1-6 alkylene, -C 1-6 alkylthioC 1-6 alkyl, -C 2-6 alkenyl, haloC 2-6 alkenyl, -C 2-6 alkynyl, -C 1-4 alkylSOC 1-4 alkyl, -C 1-4 alkylSO2C 1-4 alkyl, -SO2R 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 Rb ) q R 11 -(In the formula, R 11 C 3-7 They are cycloalkyl, 5-6 membered heterocyclyl, or 5-6 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 It is alkyl, R 13 is hydrogen, -C 1-6 It is either an alkyl group or a 5-6 membered heterocycle, or R 12 and R 13 These combine to form a 5-6 membered heterocycle, and the 5-6 membered heterocycle is either unsubstituted or alkyl-substituted. (v) 5-6 member heterocycline C 0-6 Alkyl or 5-6 member heterocyclyl (halo C) 1-4 Alkyl) (wherein each of the heterocyclyl groups is either unsubstituted or substituted with an oxo), (vi) 5-9 member heteroaryl C 0-6 Alkyl or 5-9 member heteroaryl (halo C) 1-4 Alkyl) (wherein each of the heteroaryl groups is unsubstituted or -C 1-4 Alkyl, -halo C 1-4 Alkyl, -hydroxyC 1-4 Alkylene, -C 1-4 Alkoxy, -halo C 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 3-7substituted by cycloalkyl), selected from the group consisting of, R 3a , R 3b , R 4a , R 4b , R 5a , R 5b , R 6a , and R 6b are each independently hydrogen, halo, cyano, hydroxyl, -C 1-6 alkyl, halo-C 1-6 alkyl, hydroxy-C 1-6 alkylene, -C 1-6 alkoxy, -C 1-6 alkoxy-C 1-6 alkylene, halo-C 1-6 alkoxy-C 1-6 alkylene, hydroxyhalo-C 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 together form an oxo substituent, R 7 is H, -C 1-4 alkoxy, halo, C 1-4 alkyl, or 3- to 8-membered heterocyclyl which is unsubstituted or substituted by halo, R 8 is C 3-8 cycloalkyl(C 0-6 alkyl); 4- to 6-membered heterocyclyl-C 0-6 alkyl containing 1 to 3 heteroatoms selected from N, O and S; aryl or 5- to 9-membered heteroaryl-C 0-6 alkyl containing 1 to 3 heteroatoms selected from N, O and S, and R 8is either unsubstituted or has 1 to 3 R 17 Replaced by, R 17 is halo, hydroxy, cyano, -C 1-6 Alkyl, -halo C 1-6 Alkyl, -C 1-6 Alkoxy, -halo C 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 These are, independently, hydrogen or -C 1-4 It is alkyl, R 18 is -C 1-4 Alkyl, or -C 1-4 It is a haloalkyl, p, q, r, s, and t are independently between 0 and 4.

[0009] In another embodiment, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I) or a partial thereof, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers.

[0010] In yet another aspect, the present invention provides a combination, particularly a pharmaceutically acceptable combination, comprising a therapeutically effective amount of a compound of formula (I) or a subformula thereof or a pharmaceutically acceptable salt thereof, and one or more therapeutic activators.

[0011] The compounds of the present invention can be used alone or in combination with one or more therapeutic activators to treat or prevent diseases or conditions mediated by NSD2, more specifically, diseases or conditions characterized by overexpression or undesirable upregulation of NSD2. [Modes for carrying out the invention]

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

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

[0014] When used in this specification, "-C 1-6 The term "alkyl" refers to a linear or branched hydrocarbon chain group consisting only of carbon and hydrogen atoms, not unsaturated, having 1 to 6 carbon atoms, and bonded to the rest of the molecule by single bonds. 1-4 The term "alkyl" is interpreted accordingly. 1-6 Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, and 1,1-dimethylethyl (t-butyl).

[0015] When used in this specification, "-C 2-6 The term "alkenyl" refers to a linear or branched hydrocarbon chain radical group consisting only of carbon and hydrogen atoms, containing at least one double bond, having 2 to 6 carbon atoms, and bonded to the rest of the molecule by single bonds. 2-4 The term "Alkenil" is interpreted accordingly. 2-6 Examples of alkenyls include, but are not limited to, ethenyl, prop-1-enyl, buto-1-enyl, pento-1-enyl, pento-4-enyl, and penta-1,4-dienyl.

[0016] When used in this specification, "-C 2-6 The term "alkynyl" refers to a linear or branched hydrocarbon chain radical group consisting only of carbon and hydrogen atoms, containing at least one triple bond, having 2 to 6 carbon atoms, and bonded to the rest of the molecule by single bonds. 2-4 The term "Alkinnil" is interpreted accordingly. 2-6 Examples of alkynyls include, but are not limited to, ethinyl, prop-1-inyl, buto-1-inyl, pento-1-inyl, pento-4-inyl, and penta-1,4-diinyl.

[0017] When used in this specification, "-C 1-6 The term "alkoxy" is derived from the formula -OR a It refers to the base of, and in the formula, R a Generally, C as defined above. 1-6 It is an alkyl group. 1-6 Examples of alkoxys include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, pentoxy, and hexoxy.

[0018] When used in this specification, "-C 1-6 Alkoxy C 1-6 The term "alkylene" is derived from formula -R a -OR a It refers to the base of, and in the formula, each R a This is independently of the C defined above. 1-6 It is an alkyl group. The oxygen atom can be bonded to any carbon atom in any alkyl group. C 1-6 Alkoxy C 1-6 Examples of alkyl groups include, but are not limited to, methoxymethyl, methoxyethyl, ethoxyethyl, 1-ethoxypropyl, and 2-methoxybutyl.

[0019] When used in this specification, "-C 1-4 The term "alkylcarbonyl" refers to the formula -C(=O)-R aIt refers to the base of, and in the formula, R a This is the C defined above. 1-4 It is an alkyl group.

[0020] When used in this specification, "-C 1-4 Alkylthio C 1-4 The term "alkyl" is a formula -R a -SR a It refers to the base of, and in the formula, each R a This is independently of the C defined above. 1-4 It is an alkyl group.

[0021] When used in this specification, "-hydroxyC 1-6 The term "alkylene" is C 1-6 One of the hydrogen atoms of the alkyl group is replaced by an OH group, as defined above. 1-6 It means alkyl group. Hydroxy C 1-6 Examples of alkyl groups include, but are not limited to, ethane-1-olyl, 2-methylpropane-1-olyl, hydroxymethyl, 2-hydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl, and 5-hydroxypentyl.

[0022] When used in this specification, "-aminocarbonyl C 0-6 The term "alkyl" is a formula -R a -C(=O)-NH2 refers to the group, and in the formula, R a This is a single bond or C as defined above. 1-6 It is an alkyl group.

[0023] When used in this specification, "-C 1-4 Alkylaminocarbonyl C 0-6 The term "alkyl" is a formula -R a1 -C(=O)-NH-R a2 It refers to the base of, and in the formula, R a1 This is a single bond or C as defined above. 1-6 It is an alkyl group, R a2 This is the C defined above.1-6 It is an alkyl group.

[0024] When used in this specification, "-C 1-4 Alkylaminocarbonyl C 0-6 The term "alkyl" is derived from the formula -R a1 -C(=O)-N(R a2 )-R a2 It refers to the base of, and in the formula, R a1 This is a single bond or C as defined above. 1-6 It is an alkyl group, and each R a2 This is the C defined above. 1-4 These are alkyl groups, and they can be the same or different.

[0025] When used in this specification, "-C 3-8 Cycloalkyl C 0-6 The term "alkyl" refers to a molecule consisting only of carbon and hydrogen atoms, having 3 to 8 carbon atoms and a single bond (i.e., C 3-8 By cycloalkyl) or C as defined above 1-6 This refers to a stable monocyclic saturated hydrocarbon group that is bonded to the rest of the molecule by an alkyl group. 3-7 "Cycloalkyl" and "C 3-7 Cycloalkyl C 0-4 The term "alkyl" is interpreted accordingly. 3-8 Cycloalkyl C 0-6 Examples of alkyl groups include, but are not limited to, cyclopropyl, cyclopropyl-methyl, cyclobutyl, cyclobutyl-ethyl, cyclopentyl, cyclopentyl-propyl, cyclohexyl, cycloheptaptyl, and cyclooctyl.

[0026] When used in this specification, "-C 3-8 Cycloalkoxy C 1-6 The term "alkyl" is derived from the formula -R a -OR b It refers to the base of, and in the formula, R a This is independently of the C defined above. 1-6 It is an alkyl group, R b This is the C defined above. 3-8It is a cycloalkyl compound. 3-8 Cycloalkoxy C 1-6 Examples of alkyl groups include, but are not limited to, cyclopropoxymethyl and cyclobutoxymethyl.

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

[0028] When used in this specification, "-HALO C" 1-6 The term "alkyl" refers to a C defined above, which is substituted by one or more halo groups as defined above. 1-6 Refers to an alkyl group. Halo C 1-6 Examples of alkyl groups 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" is interpreted accordingly.

[0029] When used in this specification, "-HALO C" 1-6 Alkoxy C 1-4 The term "alkylene" is derived from formula -R a1 -OR a2 It refers to the base of, and in the formula, R a1 This is the C defined above. 1-6 It is an alkyl group, R a2 This is the Halo C defined above. 1-6 It is alkyl. Halo C 1-6 Alkoxy C 1-4 Examples of alkyl groups include, but are not limited to, (difluoromethoxyl)methyl, (2,2,2-trifluoroethoxy)methyl, and (2,2-difluoroethoxy)methyl.

[0030] When used in this specification, "-hydroxylhalo C 1-6 The term "alkylene" refers to a compound C as defined above, which is substituted with one or more hydroxyl groups. 1-6Refers to an alkyl group. Hydroxylhalogen C 1-6 Examples of alkyl groups include, but are not limited to, 2,2-difluoroethane-1-olyl, 2-fluoroethane-1-olyl, and 2,2,2-trifluoroethane-1-olyl.

[0031] When used in this specification, "-CyanoC 1-6 The term "alkylene" is derived from formula -R a - Refers to the base of CN, in the formula, R a This is the C defined above. 1-4 It is an alkyl group.

[0032] When used in this specification, "-HALO C" 2-6 The term "alkenyl" refers to a C defined above, which is substituted by one or more halo groups as defined above. 2-6 This refers to the alkenyl group.

[0033] As used herein, the terms “heterocyclyl” or “heterocyclic” refer to a stable 4- to 7-membered non-aromatic monocyclic group containing one, two, or three heteroatoms individually selected from nitrogen, oxygen, and sulfur. Heterocyclyl groups may be bonded via carbon atoms or heteroatoms. The term “5- to 6-membered heterocyclyl” is interpreted accordingly. Examples of heterocyclyls include, but are not limited to, azetidinyl, oxetanyl, pyrrolinyl, pyrrolidyl, tetrahydrofuryl, tetrahydrothienyl, piperidyl, piperazinyl, tetrahydropyranil, or morpholinyl or perhydroazepinyl.

[0034] When used in this specification, "heterocyclyl C 0-6 The term "alkyl" refers to a single bond or the C defined above. 1-6 This refers to the heterocycle defined above, which is bonded to the rest of the molecule by an alkyl group.

[0035] As used herein, the term “heteroaryl” refers to a 5- to 9-membered aromatic monocyclic or fused cyclic group containing one, two, three, or four heteroatoms individually selected from nitrogen, oxygen, and sulfur. Heteroaryl groups may be bonded via carbon atoms or heteroatoms. The term “5- to 6-membered heteroaryl” is interpreted accordingly. Examples of 5- to 6-membered monocyclic heteroaryls include, but are not limited to, furyl, pyrrolyl, thienyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl, tetrazolyl, pyrazinyl, pyridadinyl, pyrimidyl, or pyridyl. Examples of condensed heteroaryls include, but are not limited to, nine-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] When used in this specification, "heteroaryl C 0-6 The term "alkyl" refers to a single bond or the C defined above. 1-6 This refers to the heteroaryl ring defined above, which is bonded to the rest of the molecule by an alkyl group.

[0037] When used herein, "heteroaryl (halo C) 1-4 The term "alkyl)" refers to the halo C defined above. 1-4 This refers to the heteroaryl ring defined above, which is bonded to the rest of the molecule by an alkyl group. Heteroaryl (halo C 1-4 An example of an alkyl group is fluoro(pyridine-2-yl)methyl.

[0038] The term "IC" as used in this specification 50 "50% inhibition" refers to the molar concentration of the inhibitor or modifier that produces 50% inhibition.

[0039] As used herein, the term "protected derivative" refers to a derivative of an inhibitor wherein a reactive site is blocked by a protecting group. Protected derivatives are useful for the preparation of inhibitors, or may themselves be active as inhibitors. Examples of protecting groups include, but are not limited to, acetyl, tetrahydropyran, methoxymethyl ether, β-methoxyethoxymethyl ether, p-methoxybenzyl, methylthiomethyl ether, pivaloyl, silyl ether, carbobenzyloxy, benzyl, tert-butoxycarbonyl, p-methoxyphenyl, 9-fluorenylmethyloxycarbonyl, acetal, ketal, acyral, dithiane, methyl ester, benzyl ester, tert-butyl ester, and silyl ester. A comprehensive list of suitable protecting groups can be found in T.W. 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 still 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 reduce or improve a disease or disorder, including those that are not recognizable to the patient (i.e., delay or suppress the onset of the disease or at least one of its clinical symptoms), or to reduce or improve at least one physical parameter or biomarker associated with the disease or disorder.

[0044] As used herein, the terms “prevent,” “preventing,” or “prevention” of any disease or disorder mean the preventive treatment of a disease or disorder, or delaying the onset or progression of a disease or disorder.

[0045] As used herein, a subject is a subject that “needs” treatment if such subject would benefit from treatment in a biological, medical, or quality of life sense.

[0046] As used herein, the term “therapeutic dose” of the compound of the present invention refers to the amount of the compound of the present invention that elicits a biological or medical response to the subject, such as reduction or inhibition of enzyme or protein activity, improvement of symptoms, alleviation of a condition, delay or stabilization of disease progression, or prevention of disease.

[0047] As used herein, the terms “antineoplastic agent” refer to therapeutic agents that are useful in 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 in the body (redness, swelling, and / or pain). Anti-inflammatory agents work by blocking certain substances in the body that cause inflammation.

[0049] As used herein, the term “pharmaceutical composition” means a compound of the present invention or a pharmaceutically acceptable salt thereof, accompanied by at least one pharmaceutically acceptable carrier, in a form suitable for oral or parenteral administration.

[0050] As used herein, the term “pharmaceutically acceptable carrier” means a substance useful for the preparation or use of a pharmaceutical composition, for example, one known to those skilled in the art (e.g., Remington, The Science and Practice of Pharmacy, 22 nd See Ed. Pharmaceutical Press, 2013, pp. 1049-1070), which include suitable diluents, solvents, dispersions, surfactants, antioxidants, preservatives, isotonic agents, buffers, emulsifiers, absorption retarders, salts, drug stabilizers, binders, excipients, disintegrants, lubricants, wetting agents, sweeteners, flavorings, dyes, and combinations thereof.

[0051] When used herein, the terms “a,” “an,” “the,” and similar terms used in the context of the present invention (particularly in the context of the claims) should be interpreted as encompassing both singular and plural forms, unless otherwise indicated herein or unless the context clearly contradicts this interpretation.

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

[0053] Various listed embodiments of the present invention are described herein. Features identified in each embodiment may be combined with other designated features to provide further embodiments of the present invention.

[0054] Embodiment 1. The compound of formula (I) described above, or its enantiomer, enantiomer mixture, or pharmaceutically acceptable salt.

[0055] Embodiment 2.A is N, the compound of formula (I) described in Embodiment 1, or its enantiomer, enantiomer mixture, or pharmaceutically acceptable salt.

[0056] Embodiment 3. R 3a However, it is hydrogen or halo, and R 3b However, hydrogen, halo, -hydroxyl, -C 1-6 It is either an alkoxy or cyano, or R 4a However, it is hydrogen or halo, and R 4b However, hydrogen, halo, -C 1-6 Alkoxy C 1-6 Alkylene, -C 1-6 Alkyl, or -HaruC 1-6 It is alkyl, or R 5a However, it is hydrogen, and R 5b However, hydrogen or -C 1-6 It is alkyl, or R 5a and R 5b A compound of formula (I) as described in Embodiment 1 or 2, or an enantiomer, enantiomer mixture thereof, or a pharmaceutically acceptable salt thereof, wherein together these compounds form an oxo substituent.

[0057] Embodiment 4. R 6a However, it is hydrogen or halo, R 6b However, hydrogen, -halo C 1-6 Alkoxy C 1-6 Alkylene, -hydroxyC 1-6 Alkylene, -hydroxyhalo C 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 However, independently, hydrogen or -C1-4 is alkyl, t is 0 to 1, or R 6a and R 6b together form an oxo substituent, the compound of formula (I) according to any one of Embodiments 1 to 3, or an enantiomer, a mixture of enantiomers, or a pharmaceutically acceptable salt thereof.

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

[0059] Embodiment 6. R 1 , R 3a , R 3b , R 4a , R 4b , R 5a , R 5b , R 6a , R 6b , and R 7 are hydrogen, the compound of formula (I) according to any one of Embodiments 1 to 5, or an enantiomer, a mixture of enantiomers, or a pharmaceutically acceptable salt thereof.

[0060] Embodiment 7. The compound of formula (I) according to Embodiment 1, wherein said compound is a compound of formula (II), **Chemical Formula** or an enantiomer, a mixture of enantiomers, or a pharmaceutically acceptable salt thereof.

[0061] Embodiment 8. The compound of formula (II) according to Embodiment 7, wherein said compound is a compound of formula (IIA), **Chemical Formula** or an enantiomer, a mixture of enantiomers, or a pharmaceutically acceptable salt thereof.

[0062] Embodiment 9.R 2 However, hydrogen, -C 1-6 Alkyl, -halo C 1-6 Alkyl, -hydroxyC 1-6 Alkylene, -hydroxyhalo C 1-6 Alkylene, -C 1-6 Alkoxy C 1-6 Alkilen, -Halo C 1-6 Alkoxy C 1-6 Alkylene, or -C 3-8 Cycloalkoxy(C 1-6 A compound according to any one of Embodiments 1 to 8, or an enantiomer thereof, an enantiomer mixture thereof, or a pharmaceutically acceptable salt thereof, which is alkyl.

[0063] Embodiment 10.R 2 However, -C 1-4 Alkylcarbonyl, -(CR a R b ) p -C(O)-OR 10 (In the formula, R 10 is hydrogen or -C 1-4 (It is alkyl.) -C(O)-(CR a R b ) q -R 11 (In the formula, R 11 However, C 3-6 They are cycloalkyl, 5-6 membered heterocyclyl, or 5-6 membered heteroaryl, each 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 (In the formula, R 12 However, hydrogen or -C 1-6 It is alkyl, R 13 However, hydrogen, -C 1-6 Alkyl, or a 5-6 membered heterocycle, R 12 and R 13 Together they form a 5-6 membered heterocycle, and the 5-6 membered heterocycle is either unsubstituted or -C 1-4 (It is substituted with alkyl) R a , R b , and R 11 However, independently, hydrogen or -C 1-4 It is alkyl, A compound according to any one of Embodiments 1 to 8, or an enantiomer thereof, an enantiomer mixture thereof, or a pharmaceutically acceptable salt thereof, wherein p, q, and r are independently 0 to 2.

[0064] Embodiment 11.R 2 but, Pyridine-2-yl, (pyridine-2-yl)methyl, or (pyridine-2-yl)ethyl (wherein the formula, the pyridine-2-yl is independently unsubstituted or 1-2-C) 1-4 Alkyl, -C 1-4 Haloalkyl, hydroxyl, -HydroxyC 1-4 Alkylene, cyano, -C 1-4 Alkoxy, -C 1-6 Alkoxy C 1-6 Alkilen, -Halo C 1-4 Alkoxy, halo, or -C 3-7 (substituted with cycloalkyl groups), Thiazol-4-yl, thiazole-2-ylmethyl, oxazol-2-yl, oxazol-2-ylmethyl, pyridazine-3-yl, pyridazine-3-ylmethyl, pyrazine-2-yl, pyrazine-2-ylmethyl, pyrimidine-4-yl, pyrimidine-4-ylmethyl (each of these independently is either unsubstituted or -C 1-4 (Substituted with alkyl or halo) 1H-pyrazole-3-yl, (1H-pyrazole-3-yl)methyl, 1-methyl-1H-pyrazole-4-yl, (1,2,3-triazole-4-yl), isoxazole-3-yl (each of these independently unsubstituted or -C1-4 Alkyl, halo, or -C 3-7 (substituted with cycloalkyl groups), 1,4-dioxane-2-yl, Pyridine-2(1H)-onyl, and A compound selected from pyridine-1-oxide-2-yl, as described in any one of Embodiments 1 to 8, or an enantiomer thereof, an enantiomer mixture thereof, or a pharmaceutically acceptable salt thereof.

[0065] Embodiment 12.R 8 The compound described in any one of Embodiments 1 to 11, or an enantiomer, enantiomer mixture, or pharmaceutically acceptable salt thereof, wherein the compound is cyclopropyl, cyclobutyl, cyclohexyl, or azetidinil.

[0066] Embodiment 13.R 8 The compound described in any one of Embodiments 1 to 11, or an enantiomer thereof, an enantiomer mixture thereof, or a pharmaceutically acceptable salt thereof, wherein the compound is 2,3-dihydrobenzofuran-5-yl, 1,3-dihydroisobenzofuran-5-yl, benzo[d][1,3]dioxol-5-yl, imidazo[1,2-a]pyridine-6-yl, or pyrazolo[1,5-a]pyridine-6-yl.

[0067] Embodiment 14.R 8 However, it is either unsubstituted or has 1-2 halos, -C 1-6 Alkyl, or -C 1-6 Pyridyl substituted with alkoxy; unsubstituted, or with one or two halos or -C 1-6 Alkyl-substituted, 1H-indazole-5-yl; or unsubstituted, or -C 1-6 A compound according to any one of Embodiments 1 to 13, or an enantiomer, enantiomer mixture, or pharmaceutically acceptable salt thereof, which is 1H-benzo[d]-imidazole-6-yl substituted with an alkyl group.

[0068] Embodiment 15.R 8However, 1 to 3 R 17 It is a phenyl substituted with, R 17 However, halo, hydroxy, cyano, -C 1-6 Alkyl, -halo C 1-6 Alkyl, -C 1-6 Alkoxy, -halo C 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 However, independently, hydrogen or -C 1-4 It is alkyl, R 18 However, -C 1-4 It is a haloalkyl, A compound according to any one of Embodiments 1 to 13, or an enantiomer thereof, an enantiomer mixture thereof, or a pharmaceutically acceptable salt thereof, wherein t is 0 to 1.

[0069] Embodiment 16.R 8 However, 1 to 3 R 17 A phenyl substituted with R 17 However, halo, hydroxy, -C 1-6 Alkyl, -halo C 1-6 Alkyl, -C 1-6 Alkoxy, or -HaruC 1-6 A compound that is an alkoxy, as described in any one of Embodiments 1 to 13, or an enantiomer thereof, an enantiomer mixture thereof, or a pharmaceutically acceptable salt thereof.

[0070] Embodiment 17. Compound of formula (II), [ka] or its enantiomer, enantiomer mixture, or pharmaceutically acceptable salt (In the formula, R 2 is hydrogen, -C 1-6 Alkyl, -halo C 1-6 Alkyl, -hydroxyC 1-6 Alkylene, -hydroxyhalo C 1-6 Alkilen, -C 1-6 Alkoxy C 1-6 Alkilen, -Halo C 1-6 Alkoxy C 1-6 Alkylene, or -C 3-8 Cycloalkoxy(C 1-6 It is alkyl, R 8 This is 1 to 3 R's 17 It is a phenyl substituted with, R 17 is, halo, -C 1-6 Alkyl, -halo C 1-6 Alkyl, -C 1-6 Alkoxy, or -halo-C 1-6 It is an alkoxy.

[0071] Embodiment 18. The compound is a compound of formula (IIA), a compound of formula (II) as described in Embodiment 17, [ka] or its enantiomer, enantiomer mixture, or pharmaceutically acceptable salt.

[0072] Embodiment 19.R 2 The compound according to any one of Embodiments 1 to 18, or an enantiomer thereof, an enantiomer mixture thereof, or a pharmaceutically acceptable salt thereof, wherein the compound is hydrogen, 2,2-difluoroethyl, 2-methylpropane-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.

[0073] Embodiment 20.R 2 but, [ka] The compound described in any one of Embodiments 1 to 18, or its enantiomer, enantiomer mixture, or pharmaceutically acceptable salt thereof.

[0074] Embodiment 21. The compound according to any one of Embodiments 1 to 20, wherein the compound is a compound of Table 2, or an enantiomer thereof, an enantiomer mixture thereof, or a pharmaceutically acceptable salt thereof.

[0075] Embodiment 22. The compound is 1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2,4,5-trifluorophenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol, 1-(3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2-(difluoromethyl)-4-fluorophenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol; 1-(3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2,5-difluoro-4-methoxyphenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol, 1-(3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2-(difluoromethyl)-4-methoxyphenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol; 1-(3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(3,5-difluoro-4-methoxyphenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol, 1-(3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(4-methoxy-2-(trifluoromethyl)phenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol, 1-(3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2-chloro-4-methoxyphenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol, 1-(3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2-(difluoromethyl)-4,5-difluorophenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol; 1-(3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2,3,4-trifluorophenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol, 1-(3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(4-fluoro-2-methoxyphenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol, and A compound selected from the group consisting of 1-(3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6'-chloro-5'-fluoro-[2,2'-bipyridine]-5-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol, according to Embodiment 21, or its enantiomer, enantiomer mixture, or pharmaceutically acceptable salt.

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

[0077] Embodiment 24. The compound according to Embodiment 1, wherein the compound is (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2,4,5-trifluorophenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol, or a pharmaceutically acceptable salt thereof. Alternatively, the compound is (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2,4,5-trifluorophenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol, or a pharmaceutically acceptable salt thereof.

[0078] Embodiment 25. The compound according to Embodiment 1, wherein the compound is (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2-(difluoromethyl)-4-fluorophenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol, or a pharmaceutically acceptable salt thereof. Alternatively, the compound is (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2-(difluoromethyl)-4-fluorophenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol, or a pharmaceutically acceptable salt thereof.

[0079] Embodiment 26. The compound according to Embodiment 1, wherein the compound is (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2,5-difluoro-4-methoxyphenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol, or a pharmaceutically acceptable salt thereof. Alternatively, the compound is (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2,5-difluoro-4-methoxyphenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol, or a pharmaceutically acceptable salt thereof.

[0080] Embodiment 27. The compound according to Embodiment 1, wherein the compound is (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2-(difluoromethyl)-4-methoxyphenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol, or a pharmaceutically acceptable salt thereof. Alternatively, the compound is (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2-(difluoromethyl)-4-methoxyphenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol, or a pharmaceutically acceptable salt thereof.

[0081] Embodiment 28. The compound according to Embodiment 1, wherein the compound is (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(3,5-difluoro-4-methoxyphenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol, or a pharmaceutically acceptable salt thereof. Alternatively, the compound is (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(3,5-difluoro-4-methoxyphenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol, or a pharmaceutically acceptable salt thereof.

[0082] Embodiment 29. The compound according to Embodiment 1, wherein the compound is (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(4-methoxy-2-(trifluoromethyl)phenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol, or a pharmaceutically acceptable salt thereof. Alternatively, the compound is (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(4-methoxy-2-(trifluoromethyl)phenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol, or a pharmaceutically acceptable salt thereof.

[0083] Embodiment 30. The compound according to Embodiment 1, wherein the compound is (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2-chloro-4-methoxyphenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol, or a pharmaceutically acceptable salt thereof. Alternatively, the compound is (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2-chloro-4-methoxyphenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol, or a pharmaceutically acceptable salt thereof.

[0084] Embodiment 31. The compound according to Embodiment 1, wherein the compound is (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2-(difluoromethyl)-4,5-difluorophenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol, or a pharmaceutically acceptable salt thereof. Alternatively, the compound is (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2-(difluoromethyl)-4,5-difluorophenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol, or a pharmaceutically acceptable salt thereof.

[0085] Embodiment 32. The compound according to Embodiment 1, wherein the compound is (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2,3,4-trifluorophenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol, or a pharmaceutically acceptable salt thereof. Alternatively, the compound is (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2,3,4-trifluorophenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol, or a pharmaceutically acceptable salt thereof.

[0086] Embodiment 33. The compound according to Embodiment 1, wherein the compound is (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(4-fluoro-2-methoxyphenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol or a pharmaceutically acceptable salt thereof. Alternatively, the compound is (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(4-fluoro-2-methoxyphenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol or a pharmaceutically acceptable salt thereof.

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

[0088] Embodiment 35. A pharmaceutical composition comprising a compound described in any one of Embodiments 1 to 34 and one or more pharmaceutically acceptable carriers.

[0089] Embodiment 36. A combination comprising one of the compounds described in any one of Embodiments 1 to 34 and one or more additional therapeutic agents.

[0090] Embodiment 37. The combination according to Embodiment 36, wherein the one or more additional therapeutic active agents are anticancer agents, analgesics, anti-inflammatory agents, or a combination thereof.

[0091] Embodiment 38. A compound according to any one of Embodiments 1 to 34, optionally combined 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 according to Embodiment 38, wherein the second therapeutic agent is an anticancer agent, an analgesic, an anti-inflammatory agent, or a combination thereof.

[0093] Embodiment 40. Use of any one of Embodiments 1 to 34, optionally combined 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 described in any one of Embodiments 1 to 34, optionally in combination with a second therapeutic agent, thereby treating the disease or condition mediated by NSD2.

[0095] Embodiment 42. A method for treating a disease or condition that benefits from or is 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 described in any one of Embodiments 1 to 34, optionally in combination with a second therapeutic agent, thereby treating the disease or condition that benefits from or is treatable by inhibition by NSD2.

[0096] Embodiment 43. The method according to 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 (especially 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 (especially acute lymphoblastic leukemia), non-Hodgkin lymphoma (especially mantle cell lymphoma), or pulmonary hypertension.

[0097] Embodiment 44. The method according to 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 lung cancer.

[0098] Embodiment 45. The method according to 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 the disease or condition that benefits from or is treatable by inhibition of NSD2, is leukemia.

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

[0101] Embodiment 48. The method according to 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 skin cancer.

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

[0103] Embodiment 50. The method according to 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 lymphoma.

[0104] Embodiment 51. The method according to Embodiment 50, wherein the lymphoma is Hodgkin lymphoma or non-Hodgkin lymphoma.

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

[0106] Embodiment 53. The method according to 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 myeloma.

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

[0108] Unless otherwise specified, the terms “compounds of the present invention” or “compound of the present invention” refer to formula (I), its subformula, and the exemplary compounds, as well as their salts, and all stereoisomers (including diastereoisomers and enantiomers), rotational isomers, tautomers, and isotopically labeled compounds (including deuterium substitutions), and uniquely formed moieties.

[0109] Depending on the selection of starting materials and procedures, compounds may exist in one form of any possible stereoisomer or as a mixture thereof, depending on the number of chiral carbon atoms, for example, as a pure optical isomer or as a mixture of stereoisomers such as a racemate and a mixture of diastereoisomers. The present invention is intended to encompass all such possible stereoisomers, including racemic mixtures, diasteriomeric mixtures, and optically pure forms. Optically active (R) and (S) stereoisomers may be prepared using chiral synthons or chiral reagents, or they may be divided using conventional techniques. Where possible, substituents on atoms with unsaturated double bonds may exist in cis-(Z) or trans-(E) configurations. If the compound contains a disubstituted cycloalkyl, the cycloalkyl substituent may have a cis or trans configuration.

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

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

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

[0113] Any resulting racemic mixture of the compound or intermediate of the present invention can be separated into optical counterparts by known methods, for example, by separation of its diastereomer salt obtained using an optically active acid or base and liberation of the optically active acid or basic compound. In particular, the basic portion can be used to separate the compound of the present invention into its optical counterparts by fractional crystallization of the salt formed using an optically active acid, for example, tartaric acid, dibenzoyl tartaric acid, diacetyl tartaric acid, di-O, O'-p-toluyl tartaric acid, mandelic acid, malic acid, or camphor-10-sulfonic acid. The racemic compound or racemic intermediate of the present invention can also be separated by chiral chromatography, for example, high-pressure liquid chromatography (HPLC) using a chiral adsorbent.

[0114] Any formulas given herein are also intended to represent both the unlabeled and isotopically labeled forms of a compound. An isotopically labeled compound has the structure represented by the formulas given herein, except that one or more atoms are replaced by atoms having a selected atomic mass or mass number. Isotopes that can be incorporated into the compounds of the present invention include, for example, isotopes of hydrogen.

[0115] Furthermore, certain isotopes, especially deuterium (i.e., 2The incorporation of H or D may result in certain therapeutic benefits due to greater metabolic stability, such as an increased half-life or reduced dose requirement in vivo, or improved therapeutic index or tolerability. In this context, deuterium is understood to be a substituent of the compound of formula (I) or its subformula. The concentration of deuterium may be defined by the isotopic enrichment factor. As used herein, the term “isotopic enrichment factor” means the ratio between the isotopic abundance and the endemic abundance of a particular isotope. Where a substituent in a compound of the present invention is indicated as deuterium, such a compound has an isotopic enrichment factor of at least 3500 (52.5% deuterium incorporation in 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) for each designated deuterium atom. It should be 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 the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, for example, respectively. 3 H, 11 C, 13 C, 14 C, 15 N, 18 F, 31 P, 32 P, 35 S, 36 Cl, 123 I, 124 I, 125 I is included. Therefore, the present invention is, for example, 3 H and 14 Radioactive isotopes such as C, or 2 H and13 It should be understood that this includes compounds that incorporate one or more of the above isotopes, including those that contain non-radioactive isotopes such as 13C. Such isotope-labeled compounds are used in metabolic studies. 14 Using C), reaction dynamics studies (for example, 2 H or 3 It is useful in detection or imaging techniques such as positron emission tomography (PET) or single-photon emission tomography (SPECT), including drug or substrate tissue distribution assays (using H), or in radiation therapy for patients. 18 F or the labeled compound may be particularly desirable for PET or SPECT studies. Isotope-labeled compounds of formula (I) or its subformulas can generally be prepared by conventional techniques known to those skilled in the art, or by a process similar to that described in the accompanying examples, using a suitable isotope-labeling reagent instead of the previously used unlabeled reagent.

[0117] The compounds of the present invention are obtained in free form or as salts thereof. As used herein, the terms “salt” or “salts” refer to acid-added or base-added salts of the compounds of the present invention. “Salt” includes, in particular, “pharmaceutically acceptable salts.” The term “pharmaceutically acceptable salt” refers to a salt that retains the biological efficacy and properties of the compounds of the present invention and is not typically biologically or otherwise undesirable. In many cases, the compounds of the present invention can form acid salts and / or base salts in the presence of amino and / or carboxyl groups or similar groups.

[0118] Pharmaceutically acceptable acid addition salts can be formed using inorganic and organic acids. Inorganic acids that can induce salt formation include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid. Organic acids that can induce salt formation 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, and sulfosalicylic acid.

[0119] pharmaceutically acceptable base addition salts can be formed using inorganic and organic bases. Inorganic bases from which salts can be derived include, for example, ammonium salts and metals from columns I to XII of the periodic table. In certain embodiments, salts are derived from sodium, potassium, ammonium, calcium, magnesium, iron, silver, zinc, and copper, with particularly preferred 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, and basic ion exchange resins. Certain organic amines include isopropylamine, benzathine, choline, diethanolamine, diethylamine, lysine, meglumine, piperazine, and tromethamine.

[0120] In another embodiment, the present invention relates to acetate, ascorbate, adipine, aspartate, benzoate, besilate, bromide / hydrobromide, bicarbonate / carbonate, bisulfate / sulfate, camphor sulfonate, caprine, chloride / hydrochloride, chlorotheophylline, citrate, ethanedisulfonate, fumarate, gluceptate, gluconate, glucuronate, glutamate, glutarate, glycolate, hippurate, hydroiodide / iodide, isethionate, lactate, lactobionate, lauryl sulfate, malate, malate The present invention provides compounds in the form of iodates, malonates, mandelates, mesylates, methylsulfates, mucate, naphthates, napsylates, nicotinates, nitrates, octadecanates, oleates, oxalates, palmitates, pamoates, phosphates / hydrogen phosphates / dihydrogen phosphates, polygalacturonates, propions, sebacinates, stearates, succinates, sulfosalicylates, sulfates, tartrates, tosylates, trifenatate, trifluoroacetates, or xinafoates.

[0121] Process for producing the compound of the present invention All methods described herein may be performed in any preferred order, unless otherwise indicated or unless otherwise clearly contradicted by the context.

[0122] The compound of formula (I) can generally be prepared as illustrated in Scheme 1, R 1 , R 2 , R 3 ... are as defined above. As shown in Scheme 1, Boc-protected 3-CN or 3-CO2Et piperidine is S in the presence of a strong base (e.g., NaHMDS) and an electrophile containing R2. N 2 or S N The Ar reaction yields compound 2 or 2' having a quaternary center, which is further hydrolyzed to the corresponding carboxylic acid 3 or primary amide 3' under either basic conditions (e.g., LiOH) or oxidative hydrolysis conditions (e.g., H2O2, NaOH). Acid 3 or amide 3' undergoes further Curtius rearrangement or Hoffmann rearrangement to produce 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 obtain coupling product 7. Ester 7 is further reduced with a mild reducing agent (e.g., NaBH4, LiCl) to benzyl alcohol 8, which undergoes a Mitsunobu reaction with Boc-protected adenine intermediate B to produce compound 9. After deprotection under acidic or hydrogenation conditions, a chiral target molecule 10 is obtained, followed by SFC separation.

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

[0124] Pharmacology and practical applications In one embodiment, the present invention provides compounds of formula (I) or a subform of the same, or pharmaceutically acceptable salts thereof, that are useful for therapeutic purposes, particularly for treating or preventing diseases or conditions mediated by NSD2.

[0125] In another aspect, the present invention provides the use of compounds of formula (I) or a subform of the same, or pharmaceutically acceptable salts thereof, for treating diseases or conditions that benefit from or are treatable by NSD2 inhibition, and for the manufacture of pharmaceuticals for treating diseases or conditions that are treatable by NSD2 inhibition.

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

[0127] Pharmaceutical composition, dosage, and administration In another embodiment, the present invention provides a pharmaceutical composition comprising a compound of the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0128] In further embodiments, the composition comprises at least two pharmaceutically acceptable carriers, such as those described herein. The pharmaceutical composition 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 may also involve inhalation or intranasal application. The pharmaceutical compositions of the present invention can be prepared 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 may be either film-coated or enteric-coated according to methods known in the art. Typically, the pharmaceutical composition is a tablet or gelatin capsule comprising an active ingredient together with one or more of the following: a) Diluents, e.g., 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, the same in the case of tablets. c) If desired, a binder, such as aluminum magnesium 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, e) Absorbents, colorants, flavorings, and sweeteners.

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

[0130] In one embodiment, the other therapeutic agent is an anticancer agent or chemotherapeutic agent. Examples of anticancer agents considered for use in combination therapy of the present invention include erlotinib, bortezomib, fulvestrant, sunitib, imatinib mesylate, letrozole, finasnate, platinum, e.g., oxaliplatin, carboplatin, and cisplatin, finasnate, fluorouracil, rapamycin, leucovorin, lapatinib, ronafamib, sorafenib, gefitinib, capumtothecin, topotecan, briostatin, adeceresin, anthracycline, carzeresin, bizeresin, and drasta Thin, auristatin, duocalmycin, eleuterobin, taxol, e.g., paclitaxel or docetaxel, cyclophasfamide, doxorubicin, vincristine, prednisone or prednisolone, other alkylating agents, e.g., mechloretamine, chlorambucil, and ifosfamide, antimetabolites, e.g., azathioprine or mercaptopurine, other microtubule inhibitors (vinca alkaloids such as vincristine, vinblastine, vinorelbine, vindesine, and taxanes), podophyllotoxin (etoposide, Teniposide, etoposide phosphate, and epipodophyllotoxin), topoisomerase inhibitors, other cytotoxins, such as actinomycin, daunorubicin, barbicin, idarubicin, edrecolomab, epirubicin, bleomycin, pricamycin, mitomycin, and other anticancer antibodies (cetuximab, bevacizumab, ibritumomab, avagovomab, adecatumumab, aftuzumab, aracizumab, alemtuzumab, anatumomab, apolizumab, bavituximab, belimumab, vivacuzumab meltansine, blinatumomab, ble Ntuximab vedotin, cantuzumab meltansine, catumazomab, cetuximab, sitatuzumab bogatox, ixutumumab, cribatuzumab tetraxetan, conatumumab, dasetuzumab, dacrizumab, detumomab, eclomeximab, edrecolomab, elotuzumab, epratuzumab, erzumaxomab, etalacizumab, farletuzumab, figtumumab, fresolimmubab, galiximab, gemtuzumab vedotin, gemtuzumab, ibritumomab tiuxetan, inotuzumab ozogamicin, intetumumab, ipilimumab,Iratumumab, Labetuzumab, Lexatuzumab, Lintuzumab, Lucatuzumab, Lumiricimab, Mapatumumab, Matsumumab, Miratuzumab, Mitsumomab, Nacolomab, Tafenatox, Naptumomab Estafenatox, Necitumumab, Nimotuzumab, Ofatumumab, Olaratumab, Oporutuzumab Monattox X, olegobomab, panitumumab, pemtumomab, pertuzumab, pintumomab, pritumumab, ramucirumab, rilotumumab, lobatumumab, rituximab, sibrotuzumab, tacutuzumab tetraxetan, tapritumomab paptox, tenatumomab, tisilinumab, tigatuzumab, tositumomab or 131 I-Tositumomab, Trastuzumab, Tremelimumab, Tuokotsuzumab, Cermolleukin, Beltuzumab, Bicilizumab, Borocicumab, Botumumab, Zaltumumab, 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 This includes, but is not limited to, lioMab-H, ING-1, anti-LCGMAbs, MT-103, KSB-303, Therex, KW2871, anti-HMI.24, anti-PTHrP, 2C4 antibody, SGN-30, TRAIL-RIMAb, prostate cancer antibody, H22xKi-r, ABX-Mai, Imteran, Monopharm-C), as well as antibody-drug conjugates containing any of the above drugs (in particular, auristatin MMAE and MMAF, mayancinoids such as DM-1, calicheamicin, or various cytotoxins).

[0131] In another embodiment, the compounds of the present invention include anastrozole (ARIMIDEX®), bicalutamide (CASODEX®), bleomycin sulfate (BLENOXANE®), busulfan (MYLERAN®), busulfan injection (BUSULFEX®), capecitabine (XELODA®), N4-pentoxycarbonyl-5-deoxy-5-fluorocytidine, carboplatin (PARAPLATIN®), carmustine (BiCNU®), and 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®), citrate Daunorubicin acid liposome injection (DAUNOXOME®), dexamethasone, docetaxel (TAXOTERE®), doxorubicin hydrochloride (ADRIAMYCIN®, RUBEX®), etoposide (VEPESID®), fludarabine phosphate (FLUDARA®), 5-fluorouracil (ADRUCIL®, EFUDEX®), flutamide (EULEXIN®), tezacitibin, 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®),Combine with another therapeutic agent selected from paclitaxel (TAXOL®), nab-paclitaxel (ABRAXANE®), Phoenix (Yttrium90 / MX-DTPA), pentostatin, polyfeprosan 20 (GLIADEL®) with carmustine implant, tamoxifen citrate (NOLVADEX®), teniposide (VUMON®), 6-thioguanine, thiotepa, tirapazamine (TIRAZONE®), topotecan hydrochloride for injection (HYCAMPTIN®), vinblastine (VELBAN®), vincristine (ONCOVIN®), and vinorelbine (NAVELBINE®).

[0132] In another embodiment, the compounds of the present invention are 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 specific embodiment, the compounds of the present invention are combined with another therapeutic agent selected from vemurafinib, debrafinib, LGX818, trametinib, MEK162, LEE011, PD-0332991, panobinostat, bellinostat, 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 compounds of the present invention is selected based on the disease or condition being treated. For example, in the treatment of melanoma, other therapeutic agents may 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 liposomes (DOXIL®, DOX-SL®, EVACET®, LIPODOX®), gemcitabine hydrochloride (GEMZAR®), topotecan hydrochloride (HYCAMTIN®), and paclitaxel (TAXOL®).

[0135] For the treatment of thyroid cancer, other therapeutic agents 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] Other therapeutic agents for the treatment of lung cancer include methotrexate, methotrexate LPF (e.g., FOLEX®, FOLEXPFS®, Abitrexate®, MEXATE®, MEXATE-AQ®), paclitaxel (TAXOL®), paclitaxel albumin-stabilized nanoparticle formulation (ABRAXANE®), afatinib dimaleate (GILOTRIF®), and pemelamine. The following drugs may be selected: trexed disodium (ALIMTA®), bevacizumab (AVASTIN®), carboplatin (PARAPLATIN®), cisplatin (PLATINOL, PLATINOL-AQ®), crizotinib (XALKORI®), elotinib 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, other therapeutic agents 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 can be selected from bosutinib (BOSULIF®), cyclophosphamide (CYTOXAN®, NEOSAR®), cytarabine (CYTOXAR-U®, TARABINE PFS®), dasatinib (SPRYCEL®), imatinib mesylate (GLEEVEC®), ponatinib (ICLUSIG®), nilotinib (TASIGNA®), and omasetaxin mepesuccinate (SYNRIBO®).

[0142] In another embodiment, the present invention provides a pharmaceutical composition comprising at least one compound of the present invention (e.g., a compound of formula (I) or a subformula thereof) or a pharmaceutically acceptable salt thereof, alone or in combination with other anticancer agents, and a pharmaceutically acceptable carrier suitable for administration to human or animal subjects.

[0143] In combination therapy, the compositions are formulated together as a combination therapeutic agent or as separate compositions. The compounds of the present invention and other therapeutic agents may 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 may be obtained from the practical version of the standard overview “The Merck Index” or from databases, e.g., Patents International (e.g., IMS World Publications). Other therapeutic agents that can be used in combination with the compounds of the present invention may be prepared and administered as described in the art, e.g., in the above document.

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

[0145] In another aspect, the present invention provides a method for treating a human or animal subject suffering from a cell proliferation disorder such as cancer, comprising administering to the subject a therapeutically effective amount of the compound of the present invention or a pharmaceutically acceptable salt thereof, alone or in combination with other anticancer agents. In combination therapy, the compound of the present invention and other anticancer agents may be administered simultaneously, concurrently, or sequentially without being limited to a specific time, and such administration provides therapeutically effective levels of the two compounds in the patient's body. Furthermore, the compound of the present invention and other therapeutic agents may be incorporated into combination therapy (i) before the combination product is made public to a physician (for example, in the case of a kit containing the compound of the present invention and other therapeutic agents), (ii) immediately before administration by the physician himself (or under the guidance of a physician), or (iii) by the patient himself, for example, during the sequential administration of the compound of the present invention and other therapeutic agents.

[0146] In one embodiment, the compounds of the present invention and other anticancer agents(s) are generally administered sequentially in any order by infusion or orally. The administration regimen may vary depending on the stage of the disease, the patient's physical condition, the safety profiles of the individual drugs, the tolerability of the individual drugs, and other criteria known to the physician(s) administering the combination. The compounds of the present invention and other anticancer agents(s) may be administered at intervals of minutes, hours, days, or even weeks from each other, depending on the specific cycle used for treatment. Furthermore, the cycle may involve administering one drug more frequently than others during the treatment cycle, and at different doses with each administration of the drugs.

[0147] In yet another embodiment, the compounds of the present invention may be combined with other anticancer agents, anti-allergic 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 and / or other anticancer agents of the present invention during or after administration. Therefore, antiallergic agents may be administered to minimize the risk of allergic reactions. Suitable antiallergic agents include corticosteroids, e.g., dexamethasone (e.g., DECADRON®), beclomethasone (e.g., BECLOVENT®), hydrocortisone (cortisone, hydrocortisone sodium phosphate, hydrocortisone sodium phosphate, e.g., also known as ALA-CORT®, hydrocortisone phosphate, Solu-CORTEF®, HYDROCORT Acetate®, and LANACORT®), prednisolone (e.g., DELTA-Cortel®, ORAPRED®, PEDIAPRED®, and PRELONE®), and prednisone (e.g., DELTASONE, LIQUID This includes 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., β-adrenergic receptor agonists, albuterol (e.g., PROVENTIL®), terbutaline (BRETHINE®)).

[0149] In other cases, patients may experience nausea during and after administration of the compounds and / or other anticancer agents of the present invention. 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 other cases, medications to alleviate pain experienced during treatment are prescribed to make the patient more comfortable. Common analgesics available without a doctor's prescription, 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] Furthermore, cytoprotective agents (neuroprotective agents, free radical scavengers, cardioprotective agents, anthracycline extravasation antagonists, nutrients, etc.) can be used as adjunctive therapies to protect normal cells from therapeutic toxicity and limit organ toxicity. Suitable cytoprotective agents include amifostine (ETHYOL®), glutamine, dimesna (TAVOCEPT®), mesna (MESNEX®), dexrazoxane (ZINECARD® or TOTECT®), xaliprodene (XAPRILA®), and leucovorin (also known as calcium leucovorin, citroborum factor, and folic acid).

[0152] In yet another embodiment, the compounds of the present invention may be used in known therapeutic processes, such as in conjunction with hormone administration or in radiotherapy. In certain examples, the compounds of the present invention may be used as radiosensitizers, particularly for the treatment of tumors that are poorly sensitive to radiotherapy.

[0153] In yet another embodiment, the present invention provides a kit comprising one or more compounds of the present invention and another therapeutic agent as described above. A typical kit comprises (a) a compound of formula (I) or a subformula thereof, or a pharmaceutically acceptable salt thereof, and (b) at least one other therapeutic agent, for example, as described above, and such a kit may further comprise other labeling, including a package insert or instructions for administration. 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 dosing intervals, or to titrate separate compositions with respect to each other, wherein at least one pharmaceutical composition comprises a compound of formula (I) or a subformula thereof.

[0154] Temperatures are expressed in Celsius. The structures of the final product, intermediates, and starting materials are confirmed by standard analytical methods, such as trace analysis and spectroscopic properties, e.g., MS, IR, NMR. The abbreviations used are those conventional in the relevant art.

[0155] All starting materials, building blocks, reagents, acids, bases, dehydrating agents, solvents, and catalysts used to synthesize the compounds of the present invention are commercially available or can be produced by any organic synthesis method known to those skilled in the art (Houben-Weyl 4th Ed. 1952, Methods of Organic Synthesis, Thiem, 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 claimed invention. Furthermore, 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. Where desired, conventional protecting groups are used to protect reactive functional groups according to standard practice; see, for example, TW Greene and PGMWuts in “Protecting Groups in Organic Synthesis”, John Wiley and Sons, 1991.

[0157] Abbreviation The abbreviations used herein are defined as follows: 1× for one pass, 2× for two passes, 3× for three passes, °C for degrees Celsius, aq for aqueous solution, 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 moles, "nM" for nanomoles, "mole" for moles, "mmol" for millimoles, "min" for minutes, "h" or "hrs" for hours, "RT" for room temperature, "ON" for overnight, "atm" for air, "psi" for pounds per square inch, "conc." for concentrates, "sat" or "sat'd" for saturation, "MW" for molecular weight, "mw" or "μwave" for microwaves, "mp" for melting point, "Wt" for weight, and "MS" or "Mass" for mass spectrometry. For electrospray ionization mass spectrometry, use "ESI"; for high resolution, use "HR"; for high resolution mass spectrometry, use "HRMS"; for liquid chromatography mass spectrometry, use "LCMS" or "LC-MS"; for high-pressure liquid chromatography, use "HPLC"; for reversed-phase HPLC, use "RP HPLC"; for thin-layer chromatography, use "TLC" or "tlc"; for nuclear magnetic resonance spectroscopy, use "NMR"; for nuclear Overhauser effect spectroscopy, use "nOe"; for protons, use " 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 enantiomer excess, as well as "α", "β", "R", "r", "S", "s", "E", and "Z" are stereochemical representations well known to those skilled in the art.

[0158] The abbreviations used in this specification below have their 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 minutes. 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 using an Agilent system (Agilent 1260 HPLC and Agilent 6130 mass spectrometer detector; column: Phenomenex Kinetex 2.6um C18, column size 4.6 × 50 mm, column temperature 40°C, gradient: 5–95% methanol in water containing 0.1% FA over 2 minutes; flow rate 2.0 mL / min (or polar gradient of 5–50% over 2 minutes, or non-polar gradient of 50–95% over 2 minutes); molecular weight scanning range of the mass spectrometer 100–1000 or 100–1500; capillary voltage 4000 V). Unless otherwise indicated, all masses are reported as the mass of the protonated parent ion.

[0161] Nuclear magnetic resonance (NMR) analysis was performed using a Bruker 400 MHz NMR spectrum analyzer. The spectral reference was either TMS or a known solvent chemical shift.

[0162] Chiral preparative HPLC method used for purification of the example SFC chiral screening was performed using the Thar Instruments Investigator system. The Thar Investigator system consists of the following: ●ALI AS Automatic Sampler ●Thar liquid delivery module (0-10 mL / 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 back pressure was set to 100 bar. Each sample was screened using a battery of 10 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 performed a gradient from 5% co-solvent to 50% co-solvent over 9 minutes, followed by a 10-minute hold in the 50% co-solvent, a switchback to the 5% co-solvent, and a 0.5-minute hold under the initial conditions. Between each gradient, there was a 4-minute equilibration method in which the 5% co-solvent was run through the next column to be screened. Typical solvents screened were MeOH, EtOH, IPA, MeOH + 0.5% NH3, EtOH + 0.5% NH3, and IPA + 0.1% NH3. Once separation is detected 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] intermediate Intermediate A: Methyl 2-(3,4-difluorophenyl)-5-fluoroisonicotinate [ka] A mixture of methyl 2-bromo-5-fluoroisonicotinate (110 g, 470.04 mmol, 1.0 equivalent), (3,4-difluorophenyl)boronic acid (111.34 g, 705.06 mmol, 1.5 equivalent), Pd(dppf)Cl2 (8.60 g, 11.75 mmol, 0.025 equivalent), and potassium carbonate (129.92 g, 940.08 mmol, 2.0 equivalent) in 1,4-dioxane (Pd(dppf)Cl2 1 L) was heated to 100°C for 16 hours under N2. The mixture was diluted with H2O (500 mL) and SiO2 (1.3 L) and stirred for 15 minutes. The organic layer was separated, washed with saline solution (500 mL), dried over Na2SO4, concentrated, and the residue was purified by combi-flash (5% to 50% ethyl acetate in PE) and grinding (PE:EA = 50 / 1, 100 mL) to obtain methyl 2-(3,4-difluorophenyl)-5-fluoroisonicotinate (intermediate A). 1 H NMR(CDCl3400MHz):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 compound 9H-purine-6-amine (15 g, 111 mmol) in THF (500 mL), di-tert-butyl dicarbonate (Boc2O) (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 hours. The THF was removed under vacuum. The residue was partitioned between SiO (400 mL) and HCl solution (1.0 M in H2O) (200 mL). The organic layer was dried over Na2SO4 and concentrated to obtain tert-butyl-6-(bis(tert-butoxycarbonyl)amino)-9H-purine-9-carboxylate (intermediate B-1).

[0167] 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) was mixed with saturated NaHCO3 solution (250 mL). The reaction mixture was stirred at 60°C for 2 hours. The MeOH was removed under vacuum. The residue was extracted with a MeOH:DCM (1:10) solution (300 mL x 7 times). The combined organic layers were washed with H2O (150 mL), dried over Na2SO4, and concentrated. The residue was washed with an ELISA:PE (1:10) solution (100 mL) and filtered. The filter cake was dried under vacuum to obtain intermediate B. 1 H NMR (400MHz, DMSO-d6) 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 the same procedure as for intermediate A. 1 H-NMR (400MHz, CDCl3) 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] A suspension of methyl 2-bromo-5-fluoroisonicotinate (1.0 g, 4.27 mmol), (3-fluoro-4-methoxyphenyl)boronic acid (1.089 g, 6.41 mmol), Cs2CO3 (2.78 g, 8.55 mmol), and PdCl2(dppf)·CH2Cl2 adduct (0.349 g, 0.427 mmol) in dioxane (10 mL) was stirred at 100 °C for 2 hours under Ar. The mixture was then diluted with water, extracted with ethyl acetate, the organic layer was dried over Na2SO4, filtered, and concentrated under vacuum. The residue was purified by CombinFlash (PE / EA, EA: 20-40%, 50 minutes) to obtain methyl 5-fluoro-2-(3-fluoro-4-methoxyphenyl)isonicotinate (intermediate D). 1 H NMR (400MHz, DMSO-d6) 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), DAST (10.41 mL, 79 mmol) was added at 0°C, and the mixture was stirred at room temperature under N2 for 18 hours. The reaction mixture was washed with water, the organic layer was dried over Na2SO4, filtered, and concentrated under vacuum. The residue was purified by CombinFlash (elution gradient: 0%~10% siRNA in n-hexane, 30 minutes) to obtain 1-bromo-2-(difluoromethyl)-4-fluorobenzene (intermediate F-1). LC-MS: [M+H] + =225.0,227.0.

[0171] A suspension of 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), KOAc (1.919 g, 19.56 mmol), and PdCl2(dppf)·CH2Cl2 (0.798 g, 0.978 mmol) adducts in dioxane (30 mL) was prepared in N The mixture was stirred at 110°C for 3 hours under a 2 atmosphere. The mixture was diluted with water, extracted with ethyl acetate, the organic layer was dried over Na2SO4, filtered, and concentrated under vacuum. The residue was purified by CombinFlash (elution gradient: 0%~10% ethyl acetate in PE, 30 minutes) to obtain 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, respectively, following the same procedure as for the preparation of intermediates F-1 and F-2.

[0173] Intermediate L-1: 1 H NMR (CDCl3,400MHz): δ 7.54-7.45 (m, 2H), 6.97-6.70 (t, J1=54.8Hz, J2=109.6Hz, 1H).

[0174] Intermediate L-2: 1H NMR (CDCl3, 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] To a solution of 6-bromo-2-chloro-3-fluoropyridine (3.0 g, 0.014 mol, 1.0 equivalent) and i-Pr3O3B (4.0 g, 0.021 mol, 1.5 equivalents) in anhydrous THF (30 mL), n-BuLi (9 mL, 0.021 mol, 2.5 M sol, 1.5 equivalents) was added dropwise at -78°C. The resulting mixture was stirred at -78°C for 1 hour and then warmed to 20°C for another 1 hour. The reaction was quenched with saturated NH4Cl (50 mL), acidified to pH=4 with concentrated HCl, and stirred at 20°C for 1 hour. Extraction was performed with ethyl acetate (50 mL x 3). The combined organic layers were dried over Na2SO4 and concentrated under vacuum to obtain 6-bromo-2-chloro-3-fluoropyridine (intermediate O-1). LC-MS: [M+H] + =176.0.

[0176] Intermediate EO was prepared by the reaction of methyl 2-bromo-5-fluoroisonicotinate with the corresponding boronic acid or ester, following the same procedure as for the preparation of intermediate A. [Table 2-1] [Table 2-2]

[0177] Intermediate 1-1: 5-fluoro-2-(3-fluoro-4-methoxyphenyl)isonicotinaldehyde [ka] A suspension of 2-bromo-5-fluoroisonicotinaldehyde (8 g, 27.5 mmol), (3-fluoro-4-methoxyphenyl)boronic acid (4.67 g, 27.5 mmol), PdCl2(dppf)·CH2Cl2 adduct (1.121 g, 1.373 mmol), and cesium carbonate (13.42 g, 41.2 mmol) in dioxane (50 mL) was stirred at 100 °C for 3 hours under a nitrogen atmosphere. The reaction mixture was filtered, and the filtrate was concentrated under vacuum to obtain a black residue. The residue was redissolved in DCM (60 mL), washed with water (20 mL) and saline solution (20 mL), dried over anhydrous magnesium sulfate, filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography (Âi in hexane and DCM, Âi / DCM / hexane = 8 / 20 / 100) to obtain 5-fluoro-2-(3-fluoro-4-methoxyphenyl)isonicotinaldehyde (intermediate 1-1). 1 H NMR(400MHz,CDCl3)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)piperidine-3-yl)carbamate [ka] To a solution of 1-(tert-butyl)3-ethylpiperidine-1,3-dicarboxylate (70.0 g, 272 mmol, 1.0 equivalent) in THF (1.4 L), LiHMDS (435 mL, 435 mmol, 1.6 equivalents, 1 M in THF) was added dropwise at -70°C under an N2 atmosphere. After stirring at -78°C for 1 hour, ((chloromethoxy)methyl)benzene (56.0 g, 354 mmol, 1.3 equivalents) was added at -78°C. The resulting mixture was warmed to 15-20°C and stirred for 16 hours. The reaction was quenched with water (800 mL) and extracted with ELISA (800 mL x 3). The combined organic layers were dried over anhydrous Na2SO4 and concentrated under vacuum. The residue was purified by Combi Flash (10% butyl in PE) to obtain 1-(tert-butyl)3-ethyl3-((benzyloxy)methyl)piperidine-1,3-dicarboxylate (22-1). 1 H NMR(400MHz,CDCl3)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-ethyl3-((benzyloxy)methyl)piperidine-1,3-dicarboxylate (22-1) (50.0 g, 132 mmol, 1.0 equivalent) and Pd / C (5.00 g, 10% wet) in dioxane / 2N HCl (1 L, v / v=1 / 1) was stirred at 20°C for 16 hours under an H2 balloon. The mixture was then filtered, and the filtrate was concentrated under vacuum. The pH of the residue was adjusted to 8-9 with aqueous NaHCO3 solution. Then, THF (500 mL) and CbzCl (45.0 g, 265 mmol, 2.0 equivalents) were added. The resulting mixture was stirred at 5-10°C for 24 hours. The mixture was extracted with ELISA (500 mL x 3). The combined organic layers were dried over anhydrous Na2SO4 and concentrated under vacuum. The residue was purified by Combi Flash (30% ELISA in PE) to obtain 1-benzyl 3-ethyl 3-(hydroxymethyl)piperidine-1,3-dicarboxylate (22-2). 1 H NMR(400MHz, CDCl3)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 equivalent) and Py (15.3 g, 193 mmol, 2.0 equivalents) in DCM (300 mL), Tf2O (57.0 g, 193 mmol, 2.0 equivalents) was added dropwise at -70°C under N2. The mixture was then warmed to 10-15°C and stirred for 1 hour. 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), dried over anhydrous Na2SO4, and concentrated under vacuum. The residue was purified by Combi Flash (10% butyl in PE) to obtain 1-benzyl 3-ethyl 3-((((trifluoromethyl)sulfonyl)oxy)methyl)piperidine-1,3-dicarboxylate (22-3). 1H NMR(400MHz,CDCl3)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 the compound cyclopropanol (3.20 g, 55.1 mmol, 1.0 equivalent) in THF (100 mL), NaH (60% in mineral oil, 2.70 g, 66.1 mmol, 1.2 equivalents) was added at 0°C. After stirring at 10-15°C for 30 minutes, 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 equivalents) in THF (100 mL) was added at 0°C. The resulting mixture was stirred at 10-15°C for 3 hours. The reaction was quenched with water (500 mL) and extracted with ELISA (500 mL x 2). The combined organic layers were washed with water (500 mL x 3), dried over anhydrous Na2SO4, and concentrated under vacuum. The residue was purified using Combi Flash (20% ELISA in PE) to obtain 1-benzyl 3-ethyl 3-(cyclopropoxymethyl)piperidine-1,3-dicarboxylate (22-4). 1 H NMR(400MHz,CDCl3)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] A mixture of 1-benzyl 3-ethyl 3-(cyclopropoxymethyl)piperidine-1,3-dicarboxylate (22-4) (10.0 g, 27.6 mmol, 1.0 equivalent) and LiOH.H2O (11.6 g, 276 mmol, 10.0 equivalent) in MeOH / THF / H2O (120 mL, v / v / v = 1 / 1 / 1) was stirred at 20°C for 16 hours. The mixture was then concentrated under vacuum to remove THF and MeOH. The residue was diluted with water (300 mL) and extracted with ELISA (200 mL). The pH of the aqueous layer was adjusted to 5 with 1N HCl, and then the aqueous layer was extracted with ELISA (300 mL x 3). The combined organic layers were dried over anhydrous Na2SO4 and concentrated under vacuum to obtain 1-((benzyloxy)carbonyl)-3-(cyclopropoxymethyl)piperidine-3-carboxylic acid (22-5). 1 H NMR(400MHz,CDCl3)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] A solution of 1-(((benzyloxy)carbonyl)-3-(cyclopropoxymethyl)piperidine-3-carboxylic acid (22-5) (6.00 g, 18 mmol, 1.0 equivalent), DPPA (5.50 g, 19.8 mmol, 1.1 equivalent), and Et3N (3.60 g, 36 mmol, 2.0 equivalent) in toluene (100 mL) was stirred at 80°C for 2 hours under N2. The reaction mixture was then concentrated under vacuum. The residue was dissolved in THF (100 mL). Then, KO H (3.03 g, 54 mmol, 3.0 equivalents) and water (10 mL) were added. The reaction mixture was stirred at 15-20°C for 4 hours. Then, Boc2O (5.90 g, 27 mmol, 1.5 equivalents) and K2CO3 (10.0 g, 72 mmol, 4.0 equivalents) were added. The reaction mixture was stirred at 15-20°C for 16 hours. The mixture was extracted with HCl (200 mL x 3). The combined organic layer was dried over anhydrous Na2SO4 and concentrated under vacuum. The residue was purified by Combi Flash (15% HCl in PE) to obtain benzyl 3-((tert-butoxycarbonyl)amino)-3-(cyclopropoxymethyl)piperidine-1-carboxylate (22-6). 1 H NMR(400MHz,CDCl3)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 stirred at 15-20°C for 1 hour under an H2 balloon. Then, Pd / C was filtered off, and the filtrate was concentrated under vacuum to obtain tert-butyl(3-(cyclopropoxymethyl)piperidine-3-yl)carbamate (intermediate 22-7). 1H NMR(400MHz,CDCl3)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)piperidine-3-yl)carbamate [ka] To a solution of 1-benzyl 3-methylpiperidine-1,3-dicarboxylate (5 g, 18.03 mmol) in THF (70 mL), 1 M LHMDS in THF (19.83 mL, 19.83 mmol) was added dropwise at -78°C for 30 minutes. The mixture was stirred at -78°C for a further 30 minutes. Then, 1,1-difluoro-2-iodoethane (6.92 g, 36.1 mmol) was added. The reaction mixture was stirred at -78°C for 2 hours. Then, the temperature was slowly raised 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), the combined organic phase was washed with water (20 mL) and saline solution (20 mL), dried over anhydrous magnesium sulfate, filtered, and concentrated. The crude product was purified by flash chromatography (elution gradient: 5%-20% phenylethylamine in hexane, 40 minutes) to obtain 1-benzyl 3-methyl 3-(2,2-difluoroethyl)piperidine-1,3-dicarboxylate (47-1). LC-MS: [M+H] + =341.9.

[0186] 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 mixed with lithium hydroxide (4.21 g, 176 mmol) in water (30 mL). The mixture was stirred overnight at room temperature to remove the solvent. The residue was diluted with water (20 mL), and the mixture was then washed with ethyl acetate (5 mL). The aqueous layer was acidified to pH=3 with HCl. The mixture was then extracted with DCM (20 mL x 3). The combined organic layers were dried over anhydrous magnesium sulfate, filtered, and concentrated under vacuum to obtain 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), TEA (4.26 mL, 30.6 mmol) and DPPA (3.70 mL, 16.80 mmol) were added. The mixture was stirred at 100°C for 2 hours under nitrogen protection. The mixture was diluted with ethyl acetate (60 mL) and washed with water (20 mL) and saline solution (60 mL). The organic phase was dried over anhydrous magnesium sulfate, filtered, and concentrated to obtain crude benzyl 3-(2,2-difluoroethyl)-3-isocyanatopiperidine-1-carboxylate (47-3). LC-MS: [M+H] + =325.1.

[0188] A solution of benzyl 3-(2,2-difluoroethyl)-3-isocyanate piperidine-1-carboxylate (47-3) (5.7 g, 12.30 mmol) in 1,4-dioxane (25 mL) was mixed with 6 M HCl (25 mL, 150 mmol). The reaction mixture was stirred at 45 °C for 30 hours. 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 basicized with sodium hydroxide to adjust the pH to 9. The mixture was then extracted with DCM (30 mL x 3). The organic phases were combined, dried over anhydrous magnesium sulfate, filtered, and concentrated to obtain 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), anhydrous Boc (4.67 mL, 20.11 mmol) and DIPEA (7.03 mL, 40.2 mmol) were added at room temperature under an N2 atmosphere. The mixture was stirred at 40°C for 20 hours. The mixture was cooled to room temperature, diluted with DCM (30 mL), washed with water (20 mL) and saline solution (20 mL), dried over anhydrous magnesium sulfate, filtered, and concentrated under vacuum to obtain the crude product. The crude product was purified by flash chromatography (elution gradient: 5%~20% siRNA in hexane, 30 minutes) to obtain benzyl 3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoroethyl)piperidine-1-carboxylate (47-5). LC-MS:[M+H] + =399.0.

[0190] To 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), 10% Pd(OH)2 wet (1.410 g, 0.502 mmol) on carbon was added, and the mixture was stirred at room temperature under a hydrogen balloon for 3 hours. The reaction mixture was filtered and concentrated to obtain crude tert-butyl(3-(2,2-difluoroethyl)piperidine-3-yl)carbamate (intermediate 47-6). LC-MS: [M+H] + =265.0.

[0191] Intermediate 78-5: Methyl tert-butyl 3-(methylcarbamoyl)piperidine-3-ylcarbamate [ka] To a solution of (NH4)2CO3 (2.13 g, 22 mmol) in H2O (8 mL), another solution of 1-benzylpiperidine-3-one (500 mg, 2 mmol) in EtOH (8 mL) was added. 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 hours. EtOH was removed under vacuum. The resulting mixture was extracted with ELISA (20 mL × 3). The organic layer was washed with H2O (20 mL) and saline solution (20 mL), dried over Na2SO4, and concentrated under vacuum. The resulting residue was recrystallized from ELISA:PE (1:5) to obtain 7-benzyl-1,3,7-triazaspiro[4.5]decane-2,4-dione (78-1). 1 H NMR(400MHz,CD3OD)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 a 10 M 2.0 M aqueous KOH solution was heated at 120°C for 16 hours. The mixture was cooled to room temperature. A concentrated HCl solution (12.0 M in H2O) was added to the mixture to adjust the pH to 4-5. H2O was removed under vacuum. The residue was washed with a MeOH:DCM solution (10%, 30 mL) and filtered. The filtrate was dried over Na2SO4 and concentrated to obtain crude 3-amino-1-benzylpiperidine-3-carboxylic acid (78-2). LC-MS[M+H] + =235.3.

[0193] A mixture of 3-amino-1-benzylpiperidine-3-carboxylic acid (78-2) (5.7 g, 24.33 mmol), K2CO3 (13.45 g, 97.31 mmol), Boc2O (8.2 mL, 36.49 mmol), THF (100 mL), and H2O (100 mL) was stirred at room temperature for 16 hours. The reaction mixture was extracted with RINKAN, the combined organic layer was washed with water and saline solution, dried over anhydrous sodium sulfate, and concentrated to obtain the residue. The residue was purified by combi-flash (silica gel, KMnO4, NH3·H2O:MeOH:DCM, 20:100:1000) to obtain 1-benzyl-3-(tert-butoxycarbonylamino)piperidine-3-carboxylic acid (78-3). LC-MS: [M+H] + =335.3.

[0194] 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 mixed with CH3NH2 (35% in MeOH, 2 mL). The mixture was stirred at room temperature for 1 hour. The reaction mixture was diluted with water and extracted with RINKAN. The combined organic phase was washed with saturated NaHCO3 aqueous solution (20 mL) and saline solution (20 mL), dried over anhydrous sodium sulfate, and concentrated to obtain the residue. The residue was purified by flash chromatography (elution gradient: 10%~50% RINKAN in PE, 30 minutes) to obtain tert-butyl1-benzyl-3-(methylcarbamoyl)piperidine-3-ylcarbamate (78-4). LC-MS: [M+H] + =348.3.

[0195] To a solution of tert-butyl 1-benzyl-3-(methylcarbamoyl)piperidine-3-ylcarbamate (78-4) (290 mg, 0.836 mmol) in EtOH (10 mL), Pd(OH)2 (87 mg) was added. The resulting mixture was stirred at room temperature under an H2 atmosphere for 4 hours. The mixture was filtered, and the filtrate was concentrated to obtain crude methyl tert-butyl 3-(methylcarbamoyl)piperidine-3-ylcarbamate (intermediate 78-5). 1 H NMR(400MHz,DMSO-d6)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)piperidine-3-yl)carbamate [ka] To a solution of tert-butyl 3-cyanopiperidine-1-carboxylate (10 g, 47.6 mmol, 1.0 equivalent) in dry THF (100 mL), 1 M LiHMDS (71.4 mL, 71.4 mmol, 1.5 equivalent) was added dropwise under nitrogen at -78°C. The mixture was stirred at -78°C for 40 minutes. Then, 3-methylbutanal (6.1 g, 71.4 mmol, 1.5 equivalent) was added all at once at -78°C. The mixture was then stirred at room temperature for 4 hours. The mixture was quenched with water (200 mL), extracted with EA (100 mL x 2), washed with saline solution (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 obtain tert-butyl 3-cyano-3-(1-hydroxy-3-methylbutyl)piperidine-1-carboxylate (89-1). 1 H NMR(400MHz,CD3OD)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] 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 equivalent) in CH2Cl2 (100 mL), DMP (11.8 g, 27.8 mmol, 1.5 equivalent) was added. The mixture was stirred at room temperature for 30 minutes. The mixture was diluted with H2O (100 mL) and extracted with CH2Cl2 (100 mL x 2). The organic layer was washed with saline solution (200 mL), dried over sodium sulfate, filtered, concentrated, and purified by combi-flash (PE / EA = 80 / 20) to obtain tert-butyl 3-cyano-3-(3-methylbutanoyl)piperidine-1-carboxylate (89-2). 1 H NMR(400MHz,CDCl3)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 equivalent) in THF (40 mL), LiHMDS (12.3 mL, 12.3 mmol, 1.1 equivalents, 1 M in THF) was added dropwise at -78°C under N2. The mixture was stirred at -78°C for 20 minutes. Then, MeI (1.8 g, 12.3 mmol, 1.1 equivalents) was added to the mixture. The mixture was stirred at room temperature for a further 4 hours. The reaction product was quenched with H2O (40 mL) and extracted with ELISA (40 mL x 2). The organic layers were combined, washed with saline solution (100 mL), dried over sodium sulfate, filtered, concentrated, and purified by combi-flash (5% toluene in hexane) to obtain tert-butyl 3-cyano-3-(2,3-dimethylbutanoyl)piperidine-1-carboxylate (89-3). 1H NMR(400MHz,CD3OD)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] A mixture of tert-butyl 3-cyano-3-(2,3-dimethylbutanoyl)piperidine-1-carboxylate (89-3) (3.3 g, 11 mmol, 1.0 equivalent) in MeOH (40 mL) was mixed with NaOH (21 mL, 21 mmol, 1 M / L) and H2O2 (17 mL, 30% aqueous solution). The mixture was stirred at room temperature for 16 hours. The mixture was quenched with saturated Na2SO3 aqueous solution (100 mL) and extracted with ELISA (100 mL x 2). The combined organic layers were washed with brine (150 mL), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography (15%~30% ELISA in PE) to obtain tert-butyl 3-carbamoyl-3-(2,3-dimethylbutanoyl)piperidine-1-carboxylate (89-4). 1 H NMR(400MHz,CDCl3)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] To a solution of tert-butyl 3-carbamoyl-3-(2,3-dimethylbutanoyl)piperidine-1-carboxylate (89-4) (1.37 g, 4.2 mmol, 1.0 equivalent) in MeOH (30 mL), PhI(OAc)2 (1.6 g, 5.04 mmol, 1.2 equivalents) and KOH (589 mg, 10.5 mmol, 2.5 equivalents) were added. The reaction mixture was stirred at 0°C for 15 hours. The mixture was stirred at room temperature for a further 2 hours. The mixture was diluted with H2O (40 mL) and extracted with ELISA (40 mL x 2). The combined organic layers were washed with saline solution (40 mL), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography (20% ethyl acetate in PE) to obtain tert-butyl 3-(2,3-dimethylbutanoyl)-3-((methoxycarbonyl)amino)piperidine-1-carboxylate (89-5). 1 H NMR(400MHz,CD3OD):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 equivalent), TFA (10 mL) was added at room temperature. The mixture was stirred at room temperature for 30 minutes. The mixture was concentrated under vacuum, diluted with H2O (15 mL), basicized to pH=11 with NH3·H2O, and extracted with CHCl3 / i-PrOH (3:1, 20 mL × 3). The combined organic layers were washed with saline solution (30 mL), dried over anhydrous Na2SO4, filtered, and concentrated to obtain methyl (3-(2,3-dimethylbutanoyl)piperidine-3-yl)carbamate (intermediate 89-6). 1H NMR(400MHz,CD3OD)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)piperidine-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), lithium chloride (10.43 g, 246 mmol) and sodium tetrahydroborate (9.31 g, 246 mmol) were added at room temperature under an N2 atmosphere, and the reaction mixture was stirred at room temperature under an N2 atmosphere for 18 hours. The reaction mixture was diluted with water and extracted with RINKAN (50 mL x 3). The combined organic phase was washed with water (50 mL) and saline solution (50 mL), dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by flash chromatography (elution gradient: 10%~50% RINKAN in PE, 30 minutes) to obtain 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), Dess-Martin periodinane (14.43 g, 34.0 mmol) was added at 0°C under an N2 atmosphere, and the reaction mixture was stirred at room temperature under an N2 atmosphere for 2 hours. The reaction mixture was diluted with 30 mL of saturated NaHCO3 aqueous solution and extracted with DCM (40 mL x 3). The combined organic phase was washed with water (50 mL) and saline solution (50 mL), dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by flash chromatography (elution gradient: 10%~40% ethyl acetate in PE, 30 minutes) to obtain benzyl 3-((tert-butoxycarbonyl)amino)-3-formylpiperidine-1-carboxylate (113-2). LC-MS:[M+H] + =362.1.

[0204] To 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 50 mL of anhydrous THF, a solution of TBAT (0.447 g, 0.828 mmol) in 15 mL of anhydrous THF was added dropwise at -78°C under an N2 atmosphere. After addition, the reaction mixture was slowly warmed to room temperature and stirred at room temperature under an N2 atmosphere for 18 hours. Then, TBAF (10.76 mL, 10.76 mmol) was added, and the reaction mixture was stirred at room temperature for a further 30 minutes. This reaction mixture was diluted with saturated NaCl solution, extracted with RINKAN (50 mL x 3), the combined organic phase was washed with water (40 mL) and saline solution (40 mL), dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by flash chromatography (elution gradient: 10%-50% ELISA in hexane, 40 minutes) to obtain 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] 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 mixed with 40 mL of HOAc / NaOAc (1:1) buffer (4 mol / L). Magnesium shavings (1.578 g, 64.9 mmol) were added gradually. The reaction mixture was stirred at room temperature for 4 hours. The reaction was quenched with water and extracted with RINKAN (30 mL x 3). The combined organic layers were washed with water (30 mL) and saline (30 mL), dried over anhydrous Na2SO4, filtered, and concentrated to obtain the residue. The residue was purified by flash chromatography (elution gradient: 0% to 40% ELISA in PE, 40 minutes) to obtain 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), Pd(OH)2 (0.068 g, 0.483 mmol) was added at room temperature, and the reaction mixture was stirred at room temperature under an H2 atmosphere for 0.5 hours. The reaction mixture was filtered and concentrated to obtain tert-butyl(3-(2,2-difluoro-1-hydroxyethyl)piperidine-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)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol [ka] A mixture of methyl 5-fluoro-2-(2,4,5-trifluorophenyl) isonicotinate (intermediate E) (2.5 g, 8.77 mmol) and 1-(3-aminopiperidine-3-yl)-2,2-difluoroethane-1-ol (intermediate 113-5A) (3.58 g, 8.77 mmol) in DMSO (40 ml) was mixed with DIPEA (40 ml, 229 mmol). The mixture was heated to 100 °C for 3 hours. The mixture was cooled to room temperature and treated with SiO (50 ml) and H₂O (50 ml). The layers were separated, and the aqueous layer was extracted with SiO (50 ml x 3). The combined organic layers were washed with saline solution, dried over Na₂SO₄, and filtered. The filtrate was concentrated. The residue was purified and eluted with ethyl(0-50%) in n-hexane to obtain methyl 5-(3-amino-3-(2,2-difluoro-1-hydroxyethyl)piperidine-1-yl)-2-(2,4,5-trifluorophenyl)isonicotinate (intermediate 156-1). 1 1H NMR (400MHz, CDCl3)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] NaBH4 (5.44 g, 144 mmol) was added at 0°C to a mixture of methyl 5-(3-amino-3-(2,2-difluoro-1-hydroxyethyl)piperidine-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). The mixture was stirred at room temperature for 4 hours. LC-MS showed that most of the SM was consumed. The mixture was quenched with H2O (50 ml) and extracted with ELISA (50 ml x 4). The combined organic layers were washed with saline, dried over Na2SO4, and filtered. The filtrate was concentrated. The filtrate was concentrated and purified by elution with MeOH in DCM (0-10%) to obtain 1-(3-amino-1-(4-(hydroxymethyl)-6-(2,4,5-trifluorophenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol (intermediate 156-2). 1 H NMR(400MHz,CDCl3)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)piperidine-1-yl)-2-(2-(difluoromethyl)-4-fluorophenyl)pyridine-4-yl)methyl)-9H-purine-6-yl)carbamate [ka] To a solution of tert-butyl(3-(2,2-difluoro-1-hydroxyethyl)piperidine-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 stirred at 80°C for 8 hours under an N2 atmosphere. The reaction mixture was diluted with water (40 mL) and extracted with ELISA (30 mL x 3). The combined organic layers were washed with water (20 mL x 3) and saline solution (20 mL), dried over anhydrous Na2SO4, filtered, and concentrated to obtain the residue. The residue was purified by flash chromatography (elution gradient: 0%~10% MeOH in DCM, 40 minutes) to obtain methyl 5-(3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidine-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)piperidine-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), NaBH4 (2.471 g, 65.3 mmol) and LiCl (2.77 g, 65.3 mmol) were added. The reaction mixture was stirred at 20°C for 2 hours under an N2 atmosphere. The reaction mixture was diluted with 30 mL of water and extracted with ELISA (20 mL x 3). The combined organic layers were washed with water (20 mL) and saline solution (20 mL), concentrated, and the crude product was obtained. The crude product was purified by flash chromatography (elution gradient: 0%~10% MeOH in DCM, 30 minutes) to obtain tert-butyl(3-(2,2-difluoro-1-hydroxyethyl)-1-(6-(2-(difluoromethyl)-4-fluorophenyl)-4-(hydroxymethyl)pyridine-3-yl)piperidine-3-yl)carbamate (intermediate 158-2). LC-MS:[M+H] + = 432.

[0211] To a solution of tert-butyl(3-(2,2-difluoro-1-hydroxyethyl)-1-(6-(2-(difluoromethyl)-4-fluorophenyl)-4-(hydroxymethyl)pyridine-3-yl)piperidine-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) in THF (50 mL), DEAD (0.550 mL, 3.48 mmol) was added, and the reaction mixture was stirred at 0°C for 30 minutes under an N2 atmosphere. The reaction mixture was diluted with 20 mL of water and extracted with ELISA (20 mL x 3). The combined organic layer was washed with water (20 mL) and saline solution (20 mL), concentrated, and the crude product was obtained. The crude product was purified by flash chromatography (elution gradient: 0%~10% MeOH in DCM, 30 minutes) to obtain tert-butyl(tert-butoxycarbonyl)(9-((5-(3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidine-1-yl)-2-(2-(difluoromethyl)-4-fluorophenyl)pyridine-4-yl)methyl)-9H-purine-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)piperidine-1-yl)-2-(2,5-difluoro-4-methoxyphenyl)pyridine-4-yl)methyl)-9H-purine-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)piperidine-3-yl)carbamate (intermediate 113-5) (2.263 g, 8.07 mmol) in DMSO (10 mL), DIPEA (23.50 mL, 135 mmol) was added under an N2 atmosphere, and the reaction mixture was stirred at 120 °C for 6 hours. The reaction mixture was diluted with water, extracted with ethyl acetate, washed with water and saline solution, dried over Na2SO4, and concentrated to obtain the crude product. The crude product was purified by flash chromatography (elution gradient: 10%-50% ethyl acetate in hexane, 30 minutes) to obtain methyl 5-(3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidine-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)piperidine-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), LiCl (0.068 g, 3.41 mmol) and NaBH4 (0.068 g, 3.41 mmol) were added at 0°C. The reaction mixture was stirred at room temperature under N2 for 2 hours. The reaction mixture was diluted with water, extracted with ELISA, washed with water and brine, dried over sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by flash chromatography (elution gradient: 10%-60% ethyl phosphate in n-hexane, 40 minutes) to obtain tert-butyl(3-(2,2-difluoro-1-hydroxyethyl)-1-(6-(2,5-difluoro-4-methoxyphenyl)-4-(hydroxymethyl)pyridine-3-yl)piperidine-3-yl)carbamate (intermediate 160-2). LC-MS: [M+H] +=530.2.

[0214] To a solution of tert-butyl(3-(2,2-difluoro-1-hydroxyethyl)-1-(6-(2,5-difluoro-4-methoxyphenyl)-4-(hydroxymethyl)pyridine-3-yl)piperidine-3-yl)carbamate (intermediate 160-2) (1.2 g, 2.3 mmol), intermediate B (0.76 g, 2.3 mmol), and PPh3 (1.783 g, 6.80 mmol) in THF (20 mL), DEAD (1.1 mL, 6.8 mmol) was added dropwise at 0°C under an N2 atmosphere, and the reaction mixture was stirred at 0°C under an N2 atmosphere for 0.5 hours. The reaction mixture was quenched with water, extracted with ethyl acetate, washed with water and saline solution, dried over Na2SO4, and concentrated to obtain the crude product. The crude product was purified by flash chromatography (elution gradient: 10%-50% ethyl acetate in n-hexane, 50 minutes) to obtain tert-butyl(tert-butoxycarbonyl)(9-((5-(3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidine-1-yl)-2-(2,5-difluoro-4-methoxyphenyl)pyridine-4-yl)methyl)-9H-purine-6-yl)carbamate (intermediate 160-3). LC-MS: [M+H]+=847.3.

[0215] Following a procedure similar to that used for the preparation of tert-butyl(3-(2,2-difluoro-1-hydroxyethyl)-1-(6-(2,5-difluoro-4-methoxyphenyl)-4-(hydroxymethyl)pyridine-3-yl)piperidine-3-yl)carbamate (intermediate 160-2), the corresponding fluoroisonicotinate was reacted with 1-(3-aminopiperidine-3-yl)-2,2-difluoroethane-1-ol (intermediate 113-5A) (intermediate numbers 162-2, 164-2, and 166-2) or tert-butyl(3-(2,2-difluoro-1-hydroxyethyl)piperidine-3-yl)carbamate (intermediate 113-5) to prepare the following intermediates (intermediate numbers 168-2, 170-2, and 172-2). [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)piperidine-1-yl)-2-(2-chloro-4-methoxyphenyl)pyridine-4-yl)methyl)-9H-purine-6-yl)carbamate [ka] To a solution of tert-butyl(1-(6-(2-chloro-4-methoxyphenyl)-4-(hydroxymethyl)pyridine-3-yl)-3-(2,2-difluoro-1-hydroxyethyl)piperidine-3-yl)carbamate (intermediate 168-2) (0.5 g, 0.947 mmol, 1.0 equivalent), intermediate B (0.318 g, 0.947 mmol, 1.1 equivalent), and n-PPh3 (0.745 g, 2.84 mmol, 3 equivalents) in THF (20 mL), DIAD (0.552 mL, 2.84 mmol, 3 equivalents) was added dropwise over 20 minutes at 0°C under N2 atmosphere. After addition, the reaction mixture was stirred at room temperature under an N2 atmosphere for 1 hour. The reaction mixture was diluted with 20 mL of water and extracted with ELISA (20 mL x 3). The combined organic layers were washed with water (20 mL) and saline solution (20 mL), concentrated, and the crude product was obtained. The crude product was purified by flash chromatography (elution gradient: 20%~70% ethyl phosphate in n-hexane, 50 minutes) to obtain tert-butyl(tert-butoxycarbonyl)(9-((5-(3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidine-1-yl)-2-(2-chloro-4-methoxyphenyl)pyridine-4-yl)methyl)-9H-purine-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)piperidine-1-yl)-2-(2-(difluoromethyl)-4,5-difluorophenyl)pyridine-4-yl)methyl)-9H-purine-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)pyridine-3-yl)piperidine-3-yl)carbamate (intermediate 170-2) (0.22 g, 0.40 mmol, 1.0 equivalent) in dry DCM (5 mL), thionyl chloride (0.146 mL, 2.02 mmol) was added dropwise at 0°C. The reaction mixture was diluted with saturated K2CO3 solution, extracted with DCM, washed with water (10 mL) and saline solution (10 mL x 2), dried over Na2SO4, and concentrated under reduced pressure to obtain tert-butyl(1-(4-(chloromethyl)-6-(2-(difluoromethyl)-4,5-difluorophenyl)pyridine-3-yl)-3-(2,2-difluoro-1-hydroxyethyl)piperidine-3-yl)carbamate (intermediate 170-3). LC-MS:[M+H] + =568.1.

[0218] Step 2. A suspension of tert-butyl(1-(4-(chloromethyl)-6-(2-(difluoromethyl)-4,5-difluorophenyl)pyridine-3-yl)-3-(2,2-difluoro-1-hydroxyethyl)piperidine-3-yl)carbamate (intermediate 170-3) (0.24 g, 0.42 mmol, 1.0 equivalent), intermediate B (0.14 g, 0.42 mmol), and K2CO3 (0.175 g, 1.27 mmol, 3.0 equivalents) in dry DMF (5 mL) was stirred at room temperature for 16 hours and quenched with water. The mixture was extracted with ethyl acetate (10 mL x 3). The combined organic layer was washed with water (10 mL x 3) and saline solution (10 mL), dried over Na2SO4, filtered, and concentrated. The crude product was purified by flash chromatography (elution gradient: 10%-70% ethyl acetate in hexane, 50 minutes) to obtain tert-butyl(tert-butoxycarbonyl)(9-(5-(3-(tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidine-1-yl)-2-(2-(difluoromethyl)-4,5-difluorophenyl)pyridine-4-yl)methyl)-9H-purine-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)piperidine-1-yl)-2-(2,3,4-trifluorophenyl)pyridine-4-yl)methyl)-9H-purine-6-yl)carbamate [ka] Following a similar procedure as described for intermediate 170-4, tert-butyl(tert-butoxycarbonyl)(9-((5-(3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidine-1-yl)-2-(2,3,4-trifluorophenyl)pyridine-4-yl)methyl)-9H-purine-6-yl)carbamate (intermediate 172-4) was obtained from tert-butyl(3-(2,2-difluoro-1-hydroxyethyl)-1-(4-(hydroxymethyl)-6-(2,3,4-trifluorophenyl)pyridine-4-yl)methyl)-9H-purine-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)piperidine-1-yl)-2-(4-fluoro-2-methoxyphenyl)pyridine-4-yl)methyl)-9H-purine-6-yl)carbamate [ka] To a solution of tert-butyl(3-(2,2-difluoro-1-hydroxyethyl)-1-(6-(4-fluoro-2-methoxyphenyl)-4-(hydroxymethyl)pyridine-3-yl)piperidine-3-yl)carbamate (intermediate 174-2) (310 mg, 0.61 mmol), intermediate B (203 mg, 0.61 mmol), and PPh3 (477 mg, 1.82 mmol) in THF (10 mL), DEAD (0.288 mL, 1.82 mmol) was added dropwise at 0°C. After addition, the reaction mixture was stirred at room temperature under an N2 atmosphere for 0.5 hours. The reaction mixture was diluted with 20 mL of water and extracted with ELISA (20 mL x 3). The combined organic layer was washed with water (20 mL) and saline (20 mL), concentrated, and the crude product was obtained. The crude product was purified by flash chromatography (elution gradient: 10%-70% ethyl acetate in PE, 50 minutes) to obtain tert-butyl(tert-butoxycarbonyl)(9-((5-(3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidine-1-yl)-2-(4-fluoro-2-methoxyphenyl)pyridine-4-yl)methyl)-9H-purine-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)piperidine-1-yl)-6'-chloro-5'-fluoro-[2,2--bipyridine]-4-yl)methyl)-9H-purine-6-yl)carbamate [ka] Following the same procedure as for intermediate 174-3, tert-butyl(tert-butoxycarbonyl)(9-((5-(3-(tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidine-1-yl)-6'-chloro-5'-fluoro-[2,2'-bipyridine]-4-yl)methyl)-9H-purine-6-yl)carbamate (intermediate 175-3). LC-MS:[M+H] + =834.1.

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

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

[0224] Intermediate 177-6: Methyl(1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridine-3-yl)-3-(6-methylpyridine-2-yl)piperidine-3-yl)carbamate

[0225] Intermediate 177-7: tert-butyl(tert-butoxycarbonyl)(9-((2-(3,4-difluorophenyl)-5-(3-((methoxycarbonyl)amino)-3-(6-methylpyridine-2-yl)piperidine-1-yl)pyridine-4-yl)methyl)-9H-purine-6-yl)carbamate [ka] To a solution of tert-butyl 3-cyanopiperidine-1-carboxylate (3.00 g, 14.3 mmol, 1.0 equivalent) and 2-fluoro-6-methylpyridine (1.66 g, 14.9 mmol, 1.05 equivalents) in THF (30 mL), KHMDS (17 mL, 17 mmol, 1.2 equivalents, 1 M in THF) was added dropwise at -70°C under an N2 atmosphere. The resulting mixture was warmed to 25°C and stirred for a further 12 hours. The reaction product was quenched with water (50 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layer was dried over anhydrous sodium 2SO4, filtered, and concentrated under vacuum. The residue was purified by combi flash (20% ethyl acetate in PE) to obtain tert-butyl 3-cyano-3-(6-methylpyridine-2-yl)piperidine-1-carboxylate (177-1). 1 H NMR(400MHz,CDCl3)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-methylpyridine-2-yl)piperidine-1-carboxylate (177-1) (2.70 g, 8.97 mmol, 1.0 equivalent) in MeOH (30 mL), NaOH (10 mL, 1 M) and H2O2 (5 mL, 30%) were added. The reaction mixture was stirred at 25°C for 12 hours, quenched with saturated Na2SO3 (100 mL), and stirred at 25°C for 1 hour. The resulting mixture was concentrated under vacuum to remove MeOH. The mixture was extracted with ELISA (1000 mL × 3). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The residue was purified with silica gel (PE / ELISA 5 / 1~1 / 1) to obtain tert-butyl 3-carbamoyl-3-(6-methylpyridine-2-yl)piperidine-1-carboxylate (177-2). 1 H NMR(400MHz,CDCl3)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-methylpyridine-2-yl)piperidine-1-carboxylate (177-2) (2.40 g, 7.51 mmol, 1.0 equivalent) and KOH (1.05 g, 18.8 mmol, 2.5 equivalents) in MeOH (30 mL), PhI(OAc)2 (2.42 g, 7.51 mmol, 1.0 equivalent) was added at 0°C. The resulting mixture was stirred at 25°C for 16 hours. The reaction was concentrated under vacuum. The residue was purified by silica gel column (PE / siRNA 10 / 1~3 / 1) to obtain tert-butyl 3-((methoxycarbonyl)amino)-3-(6-methylpyridine-2-yl)piperidine-1-carboxylate (177-3). 1H NMR(400MHz,CDCl3)δ ppm 7.55(t,J=8.0Hz,1H),7.26(d,J=8.0Hz,1H),7.01(d,J=7.6Hz,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-methylpyridine-2-yl)piperidine-1-carboxylate (177-3) (1.5 g, 4.30 mmol, 1.0 equivalent) in DCM (10 mL), HCl / dioxane (10 mL, 4 M) was added, and the resulting mixture was stirred at 25°C for 1 hour. The mixture was concentrated under vacuum to obtain methyl(3-(6-methylpyridine-2-yl)piperidine-3-yl)carbamate hydrochloride (intermediate 177-4). 1 H NMR(400MHz,CD3OD)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),3.50-3.3 9(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-methylpyridine-2-yl)piperidine-3-yl)carbamate hydrochloride (intermediate 177-4) (1.22 g, 4.27 mmol, 1.0 equivalent) in DMSO (10 mL), DIEA (3.86 g, 29.9 mmol, 7.0 equivalent) and methyl 2-(3,4-difluorophenyl)-5-fluoroisonicotinate (intermediate A) (1.14 g, 4.27 mmol, 1.0 equivalent) were added. The reaction mixture was stirred at 120°C for 3 hours. The reaction mixture was diluted with water (100 mL) and extracted with butyl (20 mL x 3). The combined organic layers were washed with saline solution (20 mL), dried over sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography (PE / EA10 / 1~3 / 1) to obtain methyl 2-(3,4-difluorophenyl)-5-(3-((methoxycarbonyl)amino)-3-(6-methylpyridine-2-yl)piperidine-1-yl) isonicotinate (177-5). 1 H NMR(400MHz,CDCl3)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] NaBH4 (914 mg, 24.2 mmol, 20 equivalents) was added to a mixture of methyl 2-(3,4-difluorophenyl)-5-(3-((methoxycarbonyl)amino)-3-(6-methylpyridine-2-yl)piperidine-1-yl) isonicotinate (177-5) (600 mg, 1.21 mmol, 1.0 equivalent) and LiCl (512 mg, 12.1 mmol, 10 equivalents) in MeOH / THF (20 mL, v / v=1 / 1). The reaction mixture was stirred at 25°C for 1 hour. The reaction product was quenched with water (50 mL) and extracted with ELISA (20 mL x 3). The combined organic layers were dried over Na2SO4 and concentrated under vacuum to obtain methyl(1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridine-3-yl)-3-(6-methylpyridine-2-yl)piperidine-3-yl)carbamate (177-6). LC-MS:[M+H] + =469.1.

[0231] DIAD (324 mg, 1.60 mmol, 1.5 equivalents) was added to a solution of methyl (1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridine-3-yl)-3-(6-methylpyridine-2-yl)piperidine-3-yl) carbamate (177-6) (500 mg, 1.07 mmol, 1.0 equivalent), intermediate B (537 mg, 1.60 mmol, 1.5 equivalents), and Bu3P (324 mg, 1.60 mmol, 1.5 equivalents) in THF (10 mL). The reaction mixture was stirred at 25°C for 3 hours. The reaction product was concentrated in a vacuum, and the residue was purified by silica gel column chromatography (eluate: PE / EA10 / 1~3:1) to obtain tert-butyl(tert-butoxycarbonyl)(9-((2-(3,4-difluorophenyl)-5-(3-((methoxycarbonyl)amino)-3-(6-methylpyridine-2-yl)piperidine-1-yl)pyridine-4-yl)methyl)-9H-purine-6-yl)carbamate (177-7). 1H NMR(400MHz,CDCl3)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-pyrazole-4-yl)piperidine-3-amine [ka] To a solution of 4-bromo-1H-pyrazole (10.0 g, 68 mmol, 1.0 equivalent) in DMF (100 mL), NaH (5.44 g, 136 mmol, 1.5 equivalents) was added at 10°C. After 30 minutes, MeI (15.0 g, 102 mmol, 2.0 equivalents) was added. The mixture was stirred at 30-35°C for 2 hours, quenched with water (200 mL), and extracted with siRNA (200 mL x 3). The combined organic layer was dried over Na2SO4 and concentrated under vacuum to obtain 4-bromo-1-methyl-1H-pyrazole (269-1). 1 H NMR (400MHz, CDCl3) 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 equivalents) in THF (140 mL), n-BuLi (29.4 mL, 72.1 mmol, 1.2 equivalents, 2.5 M in THF) was added at -70°C under N2. After 30 minutes, a solution of benzyl 3-oxopiperidine-1-carboxylate (14.0 g, 60.1 mmol, 1.0 equivalent) in THF (10 mL) was added. The reaction mixture was stirred at -70°C for 2 hours. The reaction mixture was quenched with water (200 mL) and extracted with siRNA (200 mL x 3). The combined organic layers were washed with saline, dried over Na2SO4, and concentrated under vacuum. The residue was purified by combi flash (70-90% siRNA in PE) to obtain 3-hydroxy-3-(1-methyl-1H-pyrazole-4-yl)piperidine-1-carboxylate (269-2). LC-MS: [M+H] + =316.1.

[0234] To a mixture of NaN3 (1.3 g, 19 mmol, 2.0 equivalents) in DCM (30 mL), TFA (6.00 g, 47.6 mmol, 5.0 equivalents) was added at 0°C. After 10 minutes, a solution of benzyl 3-hydroxy-3-(1-methyl-1H-pyrazole-4-yl)piperidine-1-carboxylate (intermediate 269-2) (3.00 g, 9.52 mmol, 1.0 equivalent) in DCM (10 mL) was added. The reaction mixture was stirred at 30-35°C for 16 hours. The reaction was quenched with NH3·H2O (3 mL), diluted with water (20 mL), and extracted with DCM (300 mL x 3). The combined organic layers were dried over anhydrous Na2SO4 and concentrated under vacuum. The residue was purified by Combi Flash (40% ELISA in PE) to obtain benzyl 3-azido-3-(1-methyl-1H-pyrazole-4-yl)piperidine-1-carboxylate (269-3). LC-MS: [M+H] + =341.1.

[0235] A mixture of benzyl 3-azido-3-(1-methyl-1H-pyrazole-4-yl)piperidine-1-carboxylate (269-3) (1.00 g, 2.9 mmol, 1.0 equivalent) and Pd / C (100 mg, 10% wet) in 10 mL of siRNA was stirred with an H2 balloon at 30-35°C for 16 hours. The Pd / C was filtered, and the filtrate was concentrated under vacuum to obtain 3-(1-methyl-1H-pyrazole-4-yl)piperidine-3-amine (intermediate 269-4).

[0236] Intermediate 269-6: Methyl 2-(3,4-difluorophenyl)-5-(3-((methoxycarbonyl)amino)-3-(1-methyl-1H-pyrazole-4-yl)piperidine-1-yl) isonicotinate [ka] Methyl (5-(3-amino-3-(1-methyl-1H-pyrazole-4-yl)piperidine-1-yl)-2-(3,4-difluorophenyl)isonicotinate intermediate 269-5) was prepared following the same procedure as for the preparation of intermediate 177-5 and its corresponding intermediate. 1 H NMR(400MHz,CDCl3)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] To a solution of intermediate 269-5 (500 mg, 1.17 mmol, 1.0 equivalent) and Et3N (400 mg, 3.51 mmol, 3.0 equivalents) in THF (10 mL), methyl carbonochloridate (200 mg, 1.75 mmol, 1.5 equivalents) was added at 0°C. The reaction mixture was stirred at 25-30°C for 1 hour. The reaction mixture was quenched with water (50 mL) and extracted with ethyl acetate (50 mL x 3). The combined organic layer was dried over Na2SO4 and concentrated under vacuum. The residue was purified by combi flash (60% ethyl acetate in PE) to obtain intermediate 269-6. 1 H NMR(400MHz,CDCl3)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-pyrazole-4-yl)piperidine-1-yl)pyridine-4-yl)methyl)-1H-imidazo[4,5-c]pyridine-4-yl)carbamate [ka] Methyl (1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridine-3-yl)-3-(1-methyl-1H-pyrazole-4-yl)piperidine-3-yl)carbamate (intermediate 269-7) was prepared following the same procedure as for the preparation of intermediate 177-6 and its corresponding intermediate. LC-MS:[M+H] + =458.2.

[0239] Intermediate 269-8 was prepared by replacing intermediate 177-6 with intermediate 269-7 using the same procedure as for intermediate 177-7 in Example 177. LC-MS:[M+H] + =717.4.

[0240] Intermediate 271-7: 3-(pyridine-3-yl)piperidine-3-amine [ka] To a solution of nicotinaldehyde (17.85 g, 166.7 mmol, 1.0 equivalent) in THF (250 mL) at -78°C under an N2 atmosphere, a solution of 3-butenylmagesium bromide (400 mL, 200 mmol, 0.5 M, 1.2 equivalents) was added dropwise. The reaction mixture was stirred at -78°C for 2 hours, 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 saline solution (200 mL), dried over anhydrous sodium sulfate, and concentrated under vacuum. The residue was purified by Combi-flash (30-50% EA in PE) to obtain 1-(pyridine-3-yl)pento-4-en-1-ol (271-1). 1 HNMR(CDCl3400MHz):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] To a solution of 1-(pyridine-3-yl)pento-4-en-1-ol (271-1) (10 g, 61.27 mmol, 1 equivalent) in CH2Cl2 (200 mL), Dess-Martin periodinane (38.98 g, 91.9 mmol, 1.5 equivalents) was gradually added at 16-24°C, and the suspension was stirred at 16-24°C for 30 minutes. The mixture was poured into a saturated sodium bicarbonate solution (300 mL), allowed to stand slowly, and stirred for 30 minutes. This mixture was filtered, and the aqueous layer was extracted with CH2Cl2 (100 mL x 2). The combined organic layers were washed with saline solution (100 mL), dried over anhydrous sodium sulfate, and concentrated under vacuum. The residue was purified by Combi-flash (20-30% EA in PE) to obtain 1-(pyridine-3-yl)pento-4-en-1-one (271-2). 1 HNMR(CDCl3400MHz):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-(pyridine-3-yl)pento-4-en-1-one (271-2) (8.64 g, 53.6 mmol, 1 equivalent) and 2-methylpropan-2-sulfinamide (12.99 g, 107.2 mmol, 2.0 equivalents) in anhydrous THF (120 mL), titanium ethoxide (30.57 g, 134.0 mmol, 2.5 equivalents) was added, and the reaction mixture was refluxed at 80°C for 16 hours. The mixture was cooled to room temperature, poured into saline solution (300 mL), and stirred for 30 minutes. The mixture was filtered through a Celite pad, and the aqueous layer was extracted with ethyl acetate (200 mL x 2). The combined organic layers were washed with saline solution (200 mL), dried over anhydrous sodium sulfate, and concentrated. The residue was purified by Combi-flash (20-30% EA in PE) to obtain (E)-2-methyl-N-(1-(pyridine-3-yl)pento-4-ene-1-ylidene)propan-2-sulfinamide (271-3). 1 H NMR(CDCl3400MHz):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-(pyridine-3-yl)pento-4-en-1-ylidene)propan-2-sulfinamide (271-3) (9.85 g, 37.3 mmol, 1 equivalent) in anhydrous THF (120 mL), a solution of vinyl magnesium bromide (90 mL, 90 mmol, 2.4 equivalents, 1 M in Et2O) was added dropwise, and the resulting mixture was stirred at -78°C for 2 hours. The reaction product was quenched with saturated ammonium chloride (200 mL). The mixture was extracted with ethyl acetate (100 mL x 3), the combined organic layers were washed with saline solution (100 mL), dried over anhydrous sodium sulfate, and concentrated under vacuum. The residue was purified by Combi-flash (30-70% EA in PE) to obtain 2-methyl-N-(3-(pyridine-3-yl)hepta-1,6-dien-3-yl)propan-2-sulfinamide (271-4). 1 H NMR(CDCl3400MHz):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-(pyridine-3-yl)hepta-1,6-dien-3-yl)propan-2-sulfinamide (271-4) (6.90 g, 23.6 mmol, 1.0 equivalent) in methanol (400 mL) at -78 °C was bubbling with O3 until the solution turned blue. Excess O3 was removed with N2. The mixture was concentrated under vacuum, and approximately 200 mL of methanol was removed. Diphenylmethanylamine (6.48 g, 35.4 mmol, 1.5 equivalent), sodium borohydride (3.72 g, 59.0 mmol, 2.5 equivalent), and acetic acid (1.42 g, 23.6 mmol, 1.0 equivalent) were added to the above solution, and the reaction mixture was stirred at 11-17 °C for 16 hours. The mixture was concentrated under vacuum, the residue was diluted with water (100 mL), and extracted with dichloromethane (100 mL x 3). The combined organic layers were concentrated under vacuum, and the residue was purified by Combi Flash (30-100% EA in PE) to obtain N-(1-benzhydryl-3-(pyridine-3-yl)piperidine-3-yl)-2-methylpropane-2-sulfinamide (271-6). 1 H NMR(CDCl3400MHz):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-(pyridine-3-yl)piperidine-3-yl)-2-methylpropane-2-sulfinamide (271-6) (2.34 g, 5.23 mmol, 1.0 equivalent) in TFA (24 mL), Et3SiH (4.68 mL) was added, and the reaction mixture was stirred at 80°C for 16 hours. The mixture was concentrated under vacuum, the residue was diluted with water (50 mL), and the residue was extracted with ethyl acetate (30 mL x 2) to remove impurities. The aqueous layer was neutralized with sodium hydroxide and concentrated under vacuum to obtain crude 3-(pyridine-3-yl)piperidine-3-amine (intermediate 217-7). LC-MS: [M+H] + =177.9.

[0246] Intermediate 271-9: (5-(3-amino-3-(pyridine-3-yl)piperidine-1-yl)-2-(3,4-difluorophenyl)pyridine-4-yl)methanol [ka] Methyl 5-(3-amino-3-(pyridine-3-yl)piperidine-1-yl)-2-(3,4-difluorophenyl) isonicotinate (intermediate 271-8) was prepared following the same procedure as for the preparation of intermediate 177-5 and its corresponding intermediate. 1 H NMR(CDCl3400MHz):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 by replacing intermediate 177-5 with intermediate 271-8 using the same procedure as for intermediate 177-6 in Example 177. 1 H NMR(CDCl3400MHz):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 piperidine-2-one (10 g, 100 mmol, 1.0 equivalent) in dry THF (200 mL), NaH (6 g, 60 wt%, 150 mmol, 1.5 equivalents) was gradually added below 0°C. The resulting mixture was stirred at 0°C for 0.5 hours and then at 30°C for 1 hour. Next, a solution of benzylcarbonochloride (26 g, 151.31 mmol, 1.5 equivalents) in dry THF (50 mL) was added to the suspension below 0°C for 30 minutes. The resulting mixture was stirred at 30°C for 3 hours. The reaction was quenched with saturated NH4Cl (50 mL) and extracted with ELISA (50 mL x 2). The combined organic layers were washed with saline (50 mL) and dried over Na2SO4. After filtration and concentration, the residue was purified by silica gel chromatography to obtain benzyl 2-oxopiperidine-1-carboxylate (272-1). 1H NMR (CDCl3,400MHz): 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 equivalent) in dry THF (200 mL), LDA (30 mL, 2 M, 60 mmol, 1.54 equivalents) was added under N2 conditions at -70°C for 30 minutes. After stirring at -70°C for 1 hour, methyl carbonochloride (6.23 g, 65.93 mmol, 1.69 equivalents) was added to the mixture over 30 minutes at -70°C. The resulting mixture was stirred at -70°C for 1 hour. The reaction mixture was quenched at -70°C at -70°C with saturated NH4Cl (50 mL). The mixture was extracted with ELISA (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 obtain 1-benzyl 3-methyl 2-oxopiperidine-1,3-dicarboxylate (intermediate 272-2). 1 H NMR(400MHz,CDCl3)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-methoxypyridine-3-yl)boronic acid [ka] A suspension of 3-bromo-2-methoxypyridine (9 g, 47.87 mmol, 1.0 equivalent), Mg (2.33 g, 95.73 mmol, 2.0 equivalent), and LiCl (2.54 g, 59.83 mmol, 1.25 equivalent) in dry THF (200 mL) was stirred at 35°C for 2 hours under N2. Then, B(OMe)3 (10 g, 96.23 mmol, 2.01 equivalent) was added to the reaction mixture below 0°C. The resulting mixture was then heated to 35°C with stirring for 2 hours. The reaction product was quenched with HCl (0.1 M, 200 mL) to pH=6. The mixture was extracted with RINKAN (100 mL x 3). The combined organic layers were washed with saline (100 mL) and dried over Na2SO4. After filtration and concentration, the residue was washed with chloroform (10 mL) to obtain (2-methoxypyridine-3-yl)boronic acid (intermediate 272-3). 1 H NMR(400MHz,DMSO-d6)δ ppm 8.19-8.17(dd,J1=2.0Hz,J2=4.8Hz,1H),7.89-7.88(d,J=2.0Hz,1H),3.98-6.95(dd,J1=4.8Hz,J2=6.8Hz,1H),3.87(s,3H).

[0251] Intermediate 272-5: 1-benzyl 3-methyl 3-(2-methoxypyridine-3-yl)-2-oxopiperidine-1,3-dicarboxylate [ka] To a suspension of Pb(OAc)4 (6.38 g, 14.39 mmol, 1.0 equivalent) and Hg(OAc)2 (1.17 g, 3.67 mmol, 0.26 equivalents) in chloroform (20 mL), (2-methoxypyridine-3-yl)boronic acid (272-3) (2.2 g, 14.38 mmol) (1.0 equivalent) was added under N2 conditions at 30°C. The suspension was stirred under N2 conditions at 40°C for 1 hour, and then stirred at 30°C for 16 hours. The reaction mixture of (2-methoxypyridine-3-yl)plumbanetriyl triacetate (272-4) was used directly in the next step. Next, a solution of 1-benzyl 3-methyl 2-oxopiperidine-1,3-dicarboxylate (272-2) (3.9 g, 13.23 mmol, 0.93 equivalents) in chloroform (20 mL) and pyridine (3 mL) was added to the suspension. The resulting mixture was stirred at 40°C for 12 hours under N2. LC-MS showed that intermediate 272-4 had been consumed. The reaction mixture was cooled to room temperature and filtered. The filtrate was washed with H2SO4 (1 M, 20 mL x 4) and water (20 mL x 2) and dried over Na2SO4. After filtration and concentration, the residue was purified by preparative HPLC (FA) to obtain 1-benzyl 3-methyl 3-(2-methoxypyridine-3-yl)-2-oxopiperidine-1,3-dicarboxylate (intermediate 272-5). 1 H NMR(400MHz,CDCl3)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-methoxypyridine-3-yl)piperidine-3-yl)carbamate [ka] To a solution of 1-benzyl 3-methyl 3-(2-methoxypyridine-3-yl)-2-oxopiperidine-1,3-dicarboxylate (intermediate 272-5) (1.7 g, 4.27 mmol, 1.0 equivalent) in 50 mL of dry THF, NaBH4 (0.3 g, 7.93 mmol, 1.86 equivalents) was added below 0°C. After the addition, the reaction mixture was stirred at 30°C for 2 hours. Next, Pd / C (0.45 g, 0.427 mmol, 0.1 equivalent) was added to the mixture, and the resulting mixture was stirred at 30°C for 12 hours under H2 (15 Psi). The mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain 3-(2-methoxypyridine-3-yl)piperidine-3-carboxylate (272-6).

[0253] A solution of methyl 3-(2-methoxypyridine-3-yl)piperidine-3-carboxylate (272-6) (2.2 g, 8.79 mmol, 1.0 equivalent) and Boc2O (5.75 g, 26.37 mmol, 3.0 equivalents) in MeOH (10 mL) was stirred at 30°C for 16 hours. The reaction mixture was concentrated, and the residue was purified by silica gel column chromatography to obtain 1-(tert-butyl)3-methyl 3-(2-methoxypyridine-3-yl)piperidine-1,3-dicarboxylate (272-7). 1 H NMR(400MHz,CDCl3)δ ppm 8.08-8.07(d,J=4.0Hz,1H),7.63-7.61(dd,J1=1.6Hz,J2=8.6Hz,1H),6.90-6.86(dd,J1=5.2Hz,J2=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] A solution of 1-(tert-butyl)3-(2-methoxypyridine-3-yl)piperidine-1,3-dicarboxylate (272-7) (0.85 g, 2.43 mmol, 1.0 equivalent) and LiOH (0.233 g, 9.70 mmol, 4.0 equivalents) in MeOH (10 mL) and H2O (2 mL) was heated to 40 °C with stirring for 2 hours and 4 hours at 80 °C. The reaction mixture was concentrated, and the residue was acidified to pH=3-4 with HCl (2N). 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 dried over Na2SO4. The mixture was filtered, and the filtrate was concentrated to obtain 1-(tert-butoxycarbonyl)-3-(2-methoxypyridine-3-yl)piperidine-3-carboxylic acid (272-8). 1 H NMR(400MHz,CD3OD)δppm 8.05-8.04(d,J=3.2Hz,1H),7.75-7.73(dd,J1=1.6Hz,J2=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-methoxypyridine-3-yl)piperidine-3-carboxylic acid (272-8) (0.8 g, 2.38 mmol, 1.0 equivalent) in dry toluene (20 mL), DPPA (1.31 g, 4.76 mmol, 2.0 equivalents) and TEA (0.963 g, 9.51 mmol, 4.0 equivalents) were sequentially added under N2 conditions at 28 °C. The reaction mixture was stirred at 80 °C for 12 hours. Then, MeOH (20 mL) was added to the mixture, and the mixture was stirred at 80 °C for 12 hours. The mixture was concentrated under reduced pressure, and the residue was purified by silica gel chromatography to obtain tert-butyl 3-((methoxycarbonyl)amino)-3-(2-methoxypyridine-3-yl)piperidine-1-carboxylate (272-9). 1H NMR(400MHz,CDCl3)δ ppm 8.098.074(dd,J1=2.0Hz,J2=5.2Hz,1H),7.67-7.64(dd,J1=1.6Hz,J2=7.6Hz,1H),6.92-6.88(dd,J1=4.8Hz,J2=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-methoxypyridine-3-yl)piperidine-1-carboxylate (272-9) (0.6 g, 1.64 mmol, 1.0 equivalent) in dry DCM (5 mL), TFA (1 mL) was added below 0°C. After addition, the mixture was heated to 30°C with stirring for 3 hours. The mixture was concentrated under reduced pressure to obtain methyl (3-(2-methoxypyridine-3-yl)piperidine-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-methoxypyridine-3-yl)piperidine-1-yl)pyridine-4-yl)methyl)-9H-purine-6-yl)carbamate [ka] Intermediate 272-13 was prepared following the same procedure as for the preparation of intermediate 177-7 and its corresponding intermediate. 1H NMR(400MHz,CDCl3)δ ppm 8.93(s,1H),8.50(s,1H),8.27(s,1H),8.14-8.12(dd,J1=1.6Hz,J2=5.2Hz,1H),7.77-7.75(dd,J1=1.6Hz, J2=7.6Hz,1H),7.70-7.64(m,1H),7.48-7.46(m,1H),7.20-7.13(dd,J1=5.2Hz,J2=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-pyrazole-3-yl)piperidine-1-carboxylate

[0259] Intermediate 273-5b: tert-butyl 3-cyano-3-(1-(difluoromethyl)-1H-pyrazole-5-yl)piperidine-1-carboxylate [ka] To a solution of tert-butyl 3-cyanopiperidine-1-carboxylate (25 g, 119.0 mmol, 1.0 equivalent) in THF (300 mL) at -70°C under an N2 atmosphere, LiHMDS (178 mL, 178.57 mmol, 1.5 equivalents) was added dropwise. The reaction mixture was stirred at -70°C for 30 minutes, and then CH3CHO (7.85 g, 178.57 mmol, 3 equivalents) was added. The reaction was quenched with aqueous NH4Cl solution (200 mL) and extracted with siRNA (100 mL x 3). The combined organic layer was washed with saline solution (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by Combi Flash (siRNA in PE, 10-50%) to obtain tert-butyl 3-cyano-3-(1-hydroxyethyl)piperidine-1-carboxylate (273-1). 1 H NMR(CDCl3400MHz):δ ppm 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 equivalent) in DCM (150 mL), DMP (22.5 g, 53.15 mmol, 1.5 equivalent) was added. The reaction mixture was stirred at 20°C for 1 hour. The reaction product was quenched with aqueous NaHCO3 (150 mL) and aqueous Na2SO3 (150 mL), and extracted with RINKAN (150 mL x 3). The combined organic layer was washed with aqueous NaHCO3 (150 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain crude tert-butyl 3-acetyl-3-cyanopiperidine-1-carboxylate (273-2). 1 H NMR (CDCl3400MHz): 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 equivalent) in toluene (100 mL), DMF-DMA (28.3 g, 238.1 mmol, 5.0 equivalents) was added. The reaction mixture was heated to 100 °C and stirred for 2 hours. The main product was shown by LC-MS. The reaction mixture was concentrated to obtain tert-butyl(E)-3-cyano-3-(3-(dimethylamino)acryloyl)piperidine-1-carboxylate (273-3). LC-MS: [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), hydrazine hydrate (8.85 mL, 182 mmol) was added, and the mixture was stirred at 70°C for 8 hours under an Ar atmosphere. Water (50 mL) was added, and the mixture was extracted with ELISA. The combined organic phase was work-up under aqueous conditions and concentrated under vacuum. The residue was purified by flash chromatography (PE / EA, EA: 40%, 30 minutes) to obtain tert-butyl3-cyano-3-(1H-pyrazole-3-yl)piperidine-1-carboxylate (273-4). 1 H NMR(400MHz,CDCl3)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] To a solution of tert-butyl 3-cyano-3-(1H-pyrazole-3-yl)piperidine-1-carboxylate (273-4) (500 mg, 1.809 mmol) and Cs2CO3 (2948 mg, 9.05 mmol) in DMF (10 mL), methyl 2-chloro-2,2-difluoroacetate (523 mg, 3.62 mmol) was added dropwise, 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 under vacuum, and the residue was purified by flash chromatography (PE / EA, EA: 40%, 30 minutes) to obtain tert-butyl 3-cyano-3-(1-(difluoromethyl)-1H-pyrazole-3-yl)piperidine-1-carboxylate (intermediate 273-5a) and tert-butyl 3-cyano-3-(1-(difluoromethyl)-1H-pyrazole-5-yl)piperidine-1-carboxylate (intermediate 273-5b). LC-MS:[M-100] + =227.1.

[0264] Intermediate 273-11: tert-butyltert-butoxycarbonyl(9-((5-(3-(1-(difluoromethyl)-1H-pyrazole-3-yl)-3-((methoxycarbonyl)amino)piperidine-1-yl)-2-(4-fluorophenyl)pyridine-4-yl)methyl)-9H-purine-6-yl)carbamate [ka]

[0265] Intermediate 273-11 was prepared using the same procedure as for the preparation of intermediate 177-7 and its corresponding intermediate. 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 equivalent) in THF (120 mL) was cooled to -78°C, and a solution of LiHMDS (108.18 mL, 108.18 mmol, 1.5 equivalents) was added dropwise over 20 minutes. The mixture was then stirred at -78°C for 30 minutes, followed by the addition of CH3CHO (3.81 g, 86.54 mmol, 1.2 equivalents). The reaction mixture was stirred at 32°C for 1 hour. The mixture was quenched with saturated NH4Cl solution (150 mL) and extracted with RINKAN (200 mL x 3). The combined organic phases were washed with saline solution (400 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by flash column (petroleum ether: ELISA = 9:1 to 5:1) to obtain 1-benzyl 3-methyl 3-(1-hydroxyethyl)piperidine-1,3-dicarboxylate (281-1). 1 H NMR(CDCl3400MHz):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] To a mixture of 1-benzyl 3-(1-hydroxyethyl)piperidine-1,3-dicarboxylate (281-1) (17 g, 52.9 mmol, 1.0 equivalent) in CH2Cl2 (300 mL), Dess-Martin reagent (29.2 g, 68.8 mmol, 1.3 equivalents) was gradually added over 10 minutes at 0°C. The mixture was then stirred at 32°C for 2 hours. The mixture was quenched with saturated NaHCO3 solution (300 mL), stirred for 30 minutes until the pH became 8, diluted with CH2Cl2 (300 mL), stirred for another 30 minutes, filtered, separated, and the organic phase was concentrated. The crude product was purified by flash column (petroleum ether: siRNA = 10:1) to obtain 1-benzyl 3-methyl 3-acetylpiperidine-1,3-dicarboxylate (281-2).1 H NMR (CDCl3400MHz): 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 equivalent) in MeOH (200 mL), Br2 (2.6 ml, 51.67 mmol, 1.1 equivalent) was gradually added over 5 minutes. The reaction mixture was stirred at 60°C for 1.5 hours. The mixture was concentrated, the crude product was diluted with RINKAN (300 mL), washed with saturated Na2SO3 solution (300 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 1-benzyl 3-methyl 3-(2-bromoacetyl)piperidine-1,3-dicarboxylate (281-3). LCMS:[M+H] + =399.9.

[0269] A mixture of 1-benzyl 3-methyl 3-(2-bromoacetyl)piperidine-1,3-dicarboxylate (281-3) (16 g, 32.14 mmol, 1.0 equivalent) and thiourea (4.89 g, 64.28 mmol, 2.0 equivalents) in EtOH (200 mL) was mixed with NaHCO3 (5.40 g, 64.28 mmol, 2.0 equivalents). The resulting mixture was heated at 90°C for 1.5 hours with stirring and concentrated under vacuum. The residue was dissolved in siRNA (300 mL), washed with saturated saline solution (300 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by flash column (petroleum ether:siRNA = 8:1 to 1:1) to obtain 1-benzyl 3-methyl 3-(2-aminothiazole-4-yl)piperidine-1,3-dicarboxylate (281-4). 1H NMR(CDCl3400MHz):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,1H),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] To a solution of 1-benzyl 3-methyl 3-(2-aminothiazole-4-yl)piperidine-1,3-dicarboxylate (281-4) (5 g², 13.32 mmol², 1.0 equivalent) in CH3CN (50 mL), CuCl (3.30 g, 33.29 mmol, 2.5 equivalents) was added and heated to 60°C. Subsequently, t-BuONO (3.43 g, 33.29 mmol, 2.5 equivalents) was gradually added over 5 minutes. The resulting mixture was then stirred under nitrogen at 80°C for 1 hour. The mixture was cooled to 30°C, diluted with water (50 mL), extracted with RINKAN (60 ml × 3), the organic phase was washed with saline solution (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 = 15:1~5:1) to obtain 1-benzyl 3-methyl 3-(2-chlorothiazole-4-yl)piperidine-1,3-dicarboxylate (281-5). 1 H 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] 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 equivalent) in MeOH / H2O (80 mL / 10 mL), LiOH / H2O (7.4 g, 177.27 mmol, 10.0 equivalent) was added, and the resulting mixture was stirred at 30°C for 2.0 hours. The mixture was acidified to pH 5-6 with 2 N HCl, diluted with water (40 mL), and extracted with siRNA (80 mL x 3). The combined organic phases were washed with saline solution (250 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 1-((benzyloxy)carbonyl)-3-(2-chlorothiazol-4-yl)piperidine-3-carboxylic acid (281-6). LCMS:[M+Na] + =403.0.

[0272] To 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 equivalent) and NH4Cl (1.57 g, 29.41 mmol, 2.0 equivalents) in DMF (60 mL), DIEA (7.60 g, 58.82 mmol, 4.0 equivalents) and HATU (11.18 g, 29.41 mmol, 2.0 equivalents) were added. The resulting mixture was stirred at 30°C for 1 hour, and the desired MS was observed by LC-MS. The mixture was diluted with water (40 mL) and extracted with siRNA (80 mL x 3). The combined organic phase was washed with saline solution (200 mL x 5), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by flash column (petroleum ether: Â1 = 5:1 to 1:1) to obtain benzyl 3-carbamoyl-3-(2-chlorothiazole-4-yl)piperidine-1-carboxylate (intermediate 281-7). 1H NMR(DMSO-d6400MHz):δ.7.44-7.31(m,6H),7.13(s,1H),7.01(s,1H),5.0 6(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)piperidine-1-yl)-2-(4-fluorophenyl)isonicotinate [ka] 3-(2-chlorothiazole-4-yl)-3-((methoxycarbonyl)amino)piperidine-1-carboxylate (intermediate 281-8) was prepared following the same procedure as for the preparation of intermediate 177-3 and its corresponding intermediate. 1 H NMR(CDCl3400MHz):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-chlorothiazole-4-yl)-3-((methoxycarbonyl)amino)piperidine-1-carboxylate (intermediate 281-8) (1.5 g, 3.66 mmol, 1.0 equivalent) in TFA (12.0 mL) was stirred at 66°C for 4.0 hours. The mixture was concentrated under vacuum, the residue was dissolved in Â1 (20 mL), washed with saturated NaHCO3 solution (40 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain methyl (3-(2-chlorothiazole-4-yl)piperidine-3-yl)carbamate (281-9).

[0275] DIEPA (1.87 g, 14.51 mmol, 4.0 equivalents) was added to a solution of methyl (3-(2-chlorothiazole-4-yl)piperidine-3-yl)carbamate (281-9) (1.3 g, 3.63 mmol, 1.0 equivalent) and intermediate C (903.7 mg, 3.63 mmol, 1.0 equivalent) in DMSO (20 mL). The reaction mixture was stirred at 120 °C for 12 hours. The desired MS was observed by LC-MS. The mixture was cooled to 30 °C, diluted with water (20 mL), and extracted with RINKAN (30 mL x 3). The organic phase was washed with saline solution (60 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by flash column (petroleum ether: Â15:1~6:1) to obtain methyl 5-(3-(2-chlorothiazole-4-yl)-3-((methoxycarbonyl)amino)piperidine-1-yl)-2-(4-fluorophenyl) isonicotinate (intermediate 281-10). 1 H NMR(CDCl3400MHz):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-chlorothiazole-4-yl)-3-((methoxycarbonyl)amino)piperidine-1-yl)-2-(4-fluorophenyl)pyridine-4-yl)methyl)-9H-purine-6-yl)carbamate [ka] Methyl (3-(2-chlorothiazole-4-yl)-1-(6-(4-fluorophenyl)-4-(hydroxymethyl)pyridine-3-yl)piperidine-3-yl)carbamate (intermediate 281-11) was prepared following the same procedure as for the preparation of intermediate 177-6. 1 H NMR(CDCl3400MHz):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 using the same procedure as for the preparation of intermediate 177-7 and its 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 equivalent) in THF (350 mL), LiHMDS (163 mL, 163 mmol, 1.5 equivalents) was added dropwise at -65°C under an N2 atmosphere. After stirring for 30 minutes, paraformaldehyde (9.72 g, 324 mmol, 3.0 equivalents) was added. The reaction mixture was stirred at 13-23°C for 1 hour. The reaction mixture was quenched with water (300 mL), extracted with RINKAN (300 mL x 3), and washed with saline solution (500 mL x 2). The combined organic layers were dried over Na2SO4, filtered, and concentrated under vacuum. The residue was purified by CombiFlash (20-40% ELISA in PE) to obtain 1-benzyl 3-methyl 3-(hydroxymethyl)piperidine-1,3-dicarboxylate (282-1). 1 H NMR(400MHz,CDCl3)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 equivalent) in dry DCM (150 mL), DMP (36.97 g, 87.21 mmol, 2.0 equivalents) was gradually added at 0°C. The resulting mixture was stirred at 11-20°C for 2 hours. The mixture was poured into saturated NaHCO3 solution (300 mL), extracted with DCM (300 mL x 3), washed with NaHCO3 aqueous solution (400 mL x 2), dried over Na2SO4, and concentrated. The residue was purified by flash chromatography (PE / EA = 80 / 20) to obtain 1-benzyl 3-methyl 3-formylpiperidine-1,3-dicarboxylate (282-2). 1H NMR(400MHz,CDCl3)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 equivalent) and compdimethyl(1-diazo-2-oxopropyl)phosphonate (8.17 g, 42.58 mmol, 2.0 equivalents) in dry methanol (120 mL), K2CO3 (8.8 g, 63.87 mmol, 3.0 equivalents) was added. The reaction mixture was stirred at 15-22°C for 16 hours. The mixture was concentrated. The residue was purified by flash chromatography (PE / EA = 60 / 40) to obtain 1-benzyl 3-methyl 3-ethynylpiperidine-1,3-dicarboxylate (282-3). 1 H NMR(400MHz,CDCl3)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] To a solution of 1-benzyl 3-methyl 3-ethynylpiperidine-1,3-dicarboxylate (intermediate 282-3) (4g, 13.27 mmol, 1.0 equivalent) in DMF / MeOH (40 mL / 4 mL), TMSN3 (2.3 g, 19.91 mmol, 1.5 equivalents) was added. The mixture was stirred at 100-110°C for 16 hours, diluted with water (100 mL), extracted with EA (100 mL x 4), washed with saline solution (100 mL x 3), dried over Na2SO4, filtered, and concentrated to obtain the crude compound. This was purified by CombiFlash (DCM / MeOH = 10 / 1) to obtain 1-benzyl 3-methyl 3-(2H-1,2,3-triazole-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 equivalent) in DMF (40 mL), Cs2CO3 (15.6 g, 47.9 mmol, 5 equivalents) and MeI (9.3 g, 28.74 mmol, 3 equivalents) were added. The reaction mixture was stirred at 13-18°C for 16 hours. The mixture was filtered, and the filtrate was concentrated to obtain the crude compound, which was purified by preparative HPLC to obtain 1-benzyl 3-methyl 3-(2-methyl-2H-1,2,3-triazole-4-yl)piperidine-1,3-dicarboxylate (intermediate 282-5a) and 1-benzyl 3-methyl 3-(1-methyl-1H-1,2,3-triazole-4-yl)piperidine-1,3-dicarboxylate (intermediate 282-5b).

[0284] Intermediate 282-5a: 1 H NMR(400MHz,CD3OD)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,CD3OD)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-triazole-4-yl)piperidine-3-carboxylate [ka] Intermediate 282-6 was prepared using the same procedure as for 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-triazole-4-yl)piperidine-1-carboxylate [ka] Intermediate 282-7 was prepared by replacing intermediate 281-6 with intermediate 282-6 using the same procedure as for intermediate 281-7 in Example 281. LC-MS:[M+H] + =344.1.

[0288] Intermediate 282-8: Benzyl 3-((methoxycarbonyl)amino)-3-(1-methyl-1H-1,2,3-triazole-4-yl)piperidine-1-carboxylate [ka] Intermediate 282-8 was prepared by using the same procedure as for intermediate 177-3 in Example 177, but by replacing intermediate 177-3 with intermediate 281-7. 1 H NMR(400MHz,CDCl3)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-triazole-4-yl)piperidine-3-yl)carbamate [ka] Intermediate 282-9 was prepared by replacing intermediate 269-3 with intermediate 281-8 using the same procedure as for intermediate 269-4 in Example 269. 1 H NMR(400MHz,CD3OD)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-triazole-4-yl)piperidine-1-yl) isonicotinate [ka] Intermediate 282-10 was prepared by replacing intermediate 177-4 with intermediate 282-9 using the same procedure as for intermediate 177-5 in Example 177. 1H NMR(400MHz,CDCl3)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)pyridine-3-yl)-3-(1-methyl-1H-1,2,3-triazole-4-yl)piperidine-3-yl)carbamate [ka] Intermediate 282-11 was prepared by replacing intermediate 177-5 with intermediate 282-10 using the same procedure as for intermediate 177-6 in Example 177. 1 H NMR(400MHz,CDCl3)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-triazole-4-yl)piperidine-1-yl)pyridine-4-yl)methyl)-9H-purine-6-yl)carbamate [ka] Intermediate 282-12 was prepared by replacing intermediate 177-6 with intermediate 282-11 using the same procedure as for intermediate 177-7 in Example 177. LC-MS:[M+H] + =776.3.

[0293] Intermediate: 284-4: 3-(5-methylisoxazol-3-yl)piperidine-3-amine [ka] A mixture of benzyl 3-((tert-butoxycarbonyl)amino)-3-formylpiperidine-1-carboxylate (238 mg, 0.656 mmol), hydroxylamine HCl (38 mg, 0.547 mmol), and K2CO3 (91 mg, 0.656 mmol) in EtOH / H2O (1:1, 20 mL) was stirred overnight at room temperature and treated with EA (20 mL) and H2O (10 mL). The layers were separated, and the aqueous layer was extracted with EA (10 mL x 2). The combined organic layers were washed with brine, dried over Na2SO4, and concentrated. The filtrate was concentrated to obtain benzyl(E)-3-((tert-butoxycarbonyl)amino)-3-((hydroxyimino)methyl)piperidine-1-carboxylate (284-1). LC-MS: [M+H-100] + =277.9.

[0294] 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 mixed with NCS (1.061 g, 7.95 mmol). The mixture was stirred at 40°C for 2 hours. The mixture was then treated with EA (20 mL) and H2O (20 mL). The layers were separated, and the aqueous layer was extracted with EA (20 mL x 2). The combined organic layers were washed with brine, dried over Na2SO4, and concentrated. The filtrate was concentrated to obtain crude benzyl(Z)-3-((tert-butoxycarbonyl)amino)-3-(chloro(hydroxyimino)methyl)piperidine-1-carboxylate (284-2).

[0295] 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 mixed with 2-chloroprop-1-ene (7.43 g, 97 mmol) and TEA (3.41 mL, 24.28 mmol). The mixture was heated overnight to 40°C. The mixture was then concentrated. The residue was purified and eluted with EA in n-hexane (0-50%) to obtain benzyl 3-((tert-butoxycarbonyl)amino)-3-(5-methylisoxazole-3-yl)piperidine-1-carboxylate (284-3). LC-MS: [M+H] + =415.9.

[0296] A mixture of benzyl 3-((tert-butoxycarbonyl)amino)-3-(5-methylisoxazole-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 hours. 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 dried over Na2SO4 and filtered. The filtrate was concentrated to obtain 3-(5-methylisoxazole-3-yl)piperidine-3-amine (intermediate 284-4). LC-MS: [M + H] + =182.0.

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

[0298] Intermediate 284-6 was prepared by replacing intermediate 177-5 with intermediate 284-5 using the same procedure as for intermediate 177-6 in Example 177. LC-MS:[M+H] + =400.9,401.9.

[0299] Intermediate 285-4: 3-(2-(benzyloxy)-1-fluoroethyl)piperidine-3-amine [ka] To a solution of 2-(benzyloxy)acetaldehyde (0.652 g, 4.34 mmol) in 0.025 M NaOH aqueous solution (15 mL, 0.375 mmol), 2-(benzyloxy)acetaldehyde (0.652 g, 4.34 mmol) and N,N,N-trimethylhexadecane-1-aminium chloride (0.139 g, 0.434 mmol) were added at room temperature, and the mixture was stirred at room temperature under an N2 atmosphere for 3 hours. The reaction mixture was diluted with water, extracted with ELISA (20 ml x 3), washed with water (20 ml) and saline solution (20 ml), dried over anhydrous sodium sulfate, concentrated under vacuum, and the residue was obtained. The residue was purified by flash chromatography (elution gradient: 0% to 30% siRNA in PE, 30 minutes) to obtain tert-butyl 3-(2-(benzyloxy)-1-hydroxyethyl)-3-nitropiperidine-1-carboxylate (285-1). 1 H NMR(400MHz,DMSO-d6)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), DAST (0.646 mL, 4.89 mmol) was added at 0°C, and the mixture was stirred at 0°C for 3 hours under an N2 atmosphere. The reaction mixture was then diluted with water, extracted with ELISA (20 mL x 3), and a workup was performed under aqueous conditions. The residue was obtained by concentration under vacuum. The residue was purified by flash chromatography (elution gradient: 0%~40% ELISA in PE, 30 minutes) to obtain 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), Zn powder (44.7 mg, 0.684 mmol) was added at room temperature. The mixture was then stirred at 0°C for 3 hours under an N2 atmosphere, and 4M HCl (2.281 mL, 6.84 mmol) was added dropwise at 0°C. The reaction mixture was filtered and concentrated to obtain the residue. The residue was purified by flash chromatography (elution gradient: 0%~40% siRNA in PE, 30 minutes) to obtain tert-butyl of 3-amino-3-(2-(benzyloxy)-1-fluoroethyl)piperidine-1-carboxylate (285-3). LC-MS: [M+H] + =263.2.

[0302] To a solution of 3-amino-3-(2-(benzyloxy)-1-fluoroethyl)piperidine-1-carboxylate tert-butyl (285-3) (180 mg, 0.511 mmol) in DCM (15 mL), TFA (5 mL, 64.9 mmol) was added at room temperature, and the mixture was stirred at room temperature under an N2 atmosphere for 1 hour. The mixture was then concentrated under vacuum to obtain 3-(2-(benzyloxy)-1-fluoroethyl)piperidine-3-amine (intermediate 285-4) as the TFA salt. LC-MS: [M+H] + =253.2.

[0303] Intermediate 287-6: 2,2,2-trifluoroacetaldehyde compound and N-benzyl-3-(1,2-difluoroethyl)piperidine-3-amine (1:1) [ka] AlLiH4 (13.42 g, 353.45 mmol) in THF (200 mL) was stirred at 80°C for 2 hours. The mixture was then added to 2-ethyl fluoroethyl acetate (150 g, 1.41 mol) in THF (100 mL) at -60°C and stirred at -60°C for 1 hour. The mixture was quenched with EtOH (20 mL) at -60°C, poured into a solution of ice water (500 mL) and H2SO4 (50 mL), and extracted with MTBE (150 mL x 3). The organic layer was concentrated under vacuum (25°C) to obtain the residue. The residue was purified by distillation (-0.095 MPa, 25~50°C) to obtain 1-ethoxy-2-fluoroethane-1-ol (287-1). 1 H NMR (400MHz, CD3OD) 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-fluoroethane-1-ol (287-1) (93.90 g, 868.58 mmol) and K2CO3 (12.00 g, 86.86 mmol) were added, and the mixture was stirred at 40°C for 4 hours. The mixture was concentrated under vacuum to obtain the residue. The residue was purified by column chromatography (PE / EA = 50:1 to 10:1) to obtain tert-butyl 3-(2-fluoro-1-hydroxyethyl)-3-nitropiperidine-1-carboxylate (287-2). 1 H NMR(400MHz,CDCl3)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), BAST (85.15 g, 384.87 mmol) was added and the mixture was stirred at 50°C for 4 hours. The mixture was quenched with aqueous NaHCO3 solution (1.0 L) and extracted with EA (200 mL x 3). The organic layer was concentrated under vacuum to obtain the residue. The residue was purified by column chromatography (PE:EA = 100:1 to 20:1) to obtain 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), Pd / C (5.0 g, 10% purity) was added and the mixture was stirred at 40°C for 16 hours under H2 (15 Psi). The mixture was filtered and concentrated under vacuum to obtain 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 hour. NaBH4 (5.8 g, 153.23 mmol) was added to the mixture and stirred at 40°C for 16 hours. The reaction products were detected by LC-MS. The mixture was quenched with aqueous NH4Cl (100 mL), diluted with H2O (200 mL), and extracted with EA (50 mL x 3). The organic layer was concentrated under vacuum to obtain tert-butyl 3-(benzylamino)-3-(1,2-difluoroethyl)piperidine-1-carboxylate (287-5). LC-MS: [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), TFA (10 mL) was added and the mixture was stirred at 40°C for 3 hours. The mixture was quenched with aqueous NaHCO3 (200 mL) and extracted with DCM (50 mL x 3). The organic layer was concentrated under vacuum to obtain the 2,2,2-trifluoroacetaldehyde compound and N-benzyl-3-(1,2-difluoroethyl)piperidine-3-amine (1:1) (intermediate 287-6). LCMS:[M+H] + =254.9.

[0309] Intermediate 288-9: 3-(6-methyl-1,4-dioxan-2-yl)piperidine-3-amine [ka] To a stirred solution of NaH (7.7 g, 180 mmol, 1.0 equivalent) in anhydrous THF (120 mL), prop-2-en-1-ol (10.4 g, 180 mmol, 1.0 equivalent) was added dropwise at 0°C. The reaction mixture was stirred at 0°C for 15 minutes, and then stirred at 15°C for another 15 minutes. Furthermore, 2-chloroacetic acid was added to another suspension of NaH (7.7 g, 180 mmol, 1.0 equivalent) and THF (60 mL) in anhydrous water at 0°C, stirred at 0°C for 15 minutes, then the previously prepared solution was added, and the reaction mixture was stirred at 15°C for 12 hours. The reaction mixture was quenched with water (100 mL), the aqueous layer was acidified, extracted with EA (200 mL x 3), dried over anhydrous Na2SO4, filtered, and concentrated to obtain 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 equivalent) in anhydrous DCM (60 mL), DMF (115 mg, 1.58 mmol, 0.02 equivalents) and (COCl)2 (12.1 g, 95.07 mmol, 1.2 equivalents) were added dropwise at 15°C. The reaction mixture was stirred at 15°C for 2 hours. The reaction mixture was concentrated under vacuum to obtain crude 2-(allyloxy)acetyl chloride (288-2). LC-MS: [M+C7H9N-HCl+H]+=206.2.

[0311] A solution of 1-benzyl 3-methylpiperidine-1,3-dicarboxylate (16.9 g, 60.94 mmol, 1.0 equivalent) in anhydrous THF (170 mL) was mixed dropwise with a solution of LiHMDS (61 mL, 60.94 mmol, 1.0 equivalent) at -78°C. The reaction mixture was stirred at -78°C for 1 hour, and 2-(allyloxy)acetyl chloride (288-2) (8.2 g, 60.94 mmol, 1.0 equivalent) was added dropwise at -78°C. The reaction mixture was stirred at -78°C for 1 hour, and then stirred again at 15°C for another hour. The reaction mixture was quenched with saturated NH4Cl (200 mL), extracted with EA (100 mL x 3), dried over Na2SO4, filtered, and concentrated to obtain 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 equivalent) in MeOH (100 mL), NaBH4 (2.0 g, 53.34 mmol, 1.0 equivalent) was gradually added at 15°C. The reaction mixture was stirred at 15°C for 3 hours. The reaction mixture was quenched with saturated NH4Cl (100 mL), acidified to pH 4-5 with 2N HCl, extracted with ELISA (100 ml x 3), washed with water (100 mL) and saline solution (100 mL), dried over anhydrous sodium sulfate, concentrated under vacuum to obtain the residue. The residue was purified by column chromatography (SiO2, PE / EA = 20:1 to 1:1) to obtain 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 equivalent) in dry MeCN (100 mL), NIS (10.6 g, 4.684 mmol, 1.7 equivalents) was added. The reaction mixture was stirred at 90°C for 3 hours. The reaction mixture was quenched with saturated Na2SO3 (50 mL), extracted with EA (100 mL x 3), dried over anhydrous Na2SO4, and concentrated. The residue was purified by column chromatography (SiO2, PE / EA = 20 / 1~2 / 1) to obtain 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] To 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 equivalent) and n-Bu3SnH (2.5 g, 8.54 mmol, 1.0 equivalent) in anhydrous toluene (200 mL), AIBN (140 mg, 0.85 mmol, 0.1 equivalent) was added, and the reaction mixture was stirred under N2 for 16 hours. The reaction mixture was quenched with a solution of KF (0.5 g, 8.54 mmol, 1.0 equivalent) in H2O (50 mL), diluted with water (100 mL), extracted with EA (100 mL x 3), dried over anhydrous Na2SO4, and concentrated under vacuum to obtain the residue. The residue was purified by column chromatography (SiO2, PE / EA = 20 / 1 to 1 / 1) to obtain 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] 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 equivalent) in MeOH (50 mL) was added to a solution of LiOH·H2O (4.0 g, 95.38 mmol, 10.0 equivalent) in H2O (50 mL) at room temperature. The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was concentrated under vacuum to remove most of the MeOH, diluted with 20 mL of water, and the aqueous phase was washed with EA (10 mL x 3). Next, the aqueous phase was acidified to pH 3-4 with 2N HCl, extracted with EA (50 mL x 3), dried with anhydrous Na2SO4, and concentrated under vacuum to obtain 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 equivalent) in anhydrous THF (800 mL), TEA (1.7 g, 17.34 mmol, 3.0 equivalents) and DPPA (2.2 g, 8.09 mmol, 1.4 equivalents) were added at 20°C. The reaction mixture was stirred at 70°C for 2 hours, cooled to 20°C, and then a solution of KOH (1.0 g, 17.34 mmol, 3.0 equivalents) in H2O (10 mL) was added. The reaction mixture was stirred at 20°C for 2 hours. The reaction mixture was concentrated, extracted with EA (50 mL x 3), and washed with water (50 mL) and saline solution (50 mL). The organic layer was dried over Na2SO4 and concentrated under vacuum. The crude product was purified by column chromatography (SiO2, PE / EA = 20 / 1~EA) to obtain a non-pure product, which was further purified by preparative HPLC to obtain 3-amino-3-(6-methyl-1,4-dioxan-2-yl)piperidine-1-carboxylate benzyl (288-8). 1H NMR (400MHz, CDCl3) 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 equivalent) in MeOH (10 mL), Pd(OH)2 (104 mg, 10 wt%) was added, and the reaction mixture was stirred at 20°C for 8 hours under an H2 atmosphere (16 Psi). The reaction mixture was filtered and concentrated under vacuum to obtain crude 3-(6-methyl-1,4-dioxan-2-yl)piperidine-3-amine (intermediate 288-9).

[0318] Intermediate 292-3: 3-oxa-1,7-diazaspiro[4.5]decane-2-one [ka] A solution of 1-benzyl 3-ethyl 3-(hydroxymethyl)piperidine-1,3-dicarboxylate (intermediate 22-2) (2.0 g, 6.97 mmol, 1.0 equivalent) in THF / MeOH / H2O (1:1:1, 30 mL) is mixed with LiOH . H2O (0.87 g, 20.9 mmol, 3.0 equivalents) was added at 20°C. The mixture was stirred at 20°C for 8 hours. Water (50 mL) was added, and the mixture was extracted with ethyl acetate (100 mL x 3). The aqueous phase was acidified to pH 2-3 with 1 M HCl, and then extracted with ethyl acetate (100 mL x 3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 1-(tert-butoxycarbonyl)-3-(hydroxymethyl)piperidine-3-carboxylate (292-1). 1H NMR(CDCl3400MHz):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 equivalent) in toluene (40 mL), TEA (1.4 g, 13.9 mmol, 2.0 equivalents) and DPPA (2.7 g, 10.4 mmol, 1.5 equivalents) were added under N2 at 20°C. The reaction mixture was then stirred at 110°C for 8 hours. H2O (100 mL) was added, and the mixture was extracted with ELISA (100 mL × 3). The combined organic layer was washed with saline solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 1:1) to obtain tert-butyl 2-oxo-3-oxa-1,7-diazaspiro[4.5]decane-7-carboxylate (292-2). 1 H NMR(CDCl3400MHz):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 equivalent) in HCl / dioxane (5 mL, 4 N) was stirred at 20°C for 2 hours. The mixture was concentrated to obtain crude 3-oxa-1,7-diazaspiro[4.5]decane-2-one (intermediate 292-3).

[0321] Intermediate 293-8: 3-(1,1-difluoroprop-1-en-2-yl)piperidine-3-amine [ka] To a solution of tert-butyl 3-cyanopiperidine-1-carboxylate (25 g, 119.0 mmol, 1.0 equivalent) in THF (300 mL) at -70°C under an N2 atmosphere, LiHMDS (178 mL, 178.57 mmol, 1.5 equivalents) was added dropwise. The reaction mixture was stirred at -70°C for 30 minutes, and then CH3CHO (7.85 g, 178.57 mmol, 3 equivalents) was added. The reaction mixture was quenched with aqueous NH4Cl solution (200 mL) and extracted with siRNA (100 mL x 3). The combined organic layer was washed with saline solution (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by Combi-flash (siRNA in PE, 10%-50%) to obtain 3-(1-hydroxyethyl)-3-methylpiperidine-1-carboxylate tert-butyl (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 equivalent) in DCM (150 mL), DMP (22.5 g, 53.15 mmol, 1.5 equivalent) was added. The reaction mixture was stirred at 20°C for 1 hour. The reaction product was quenched with aqueous NaHCO3 (150 mL) and aqueous Na2SO3 (150 mL) and extracted with RINKAN (150 mL x 3). The combined organic layer was washed with aqueous NaHCO3 (150 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain tert-butyl 3-acetyl-3-cyanopiperidine-1-carboxylate (293-2). 1 H NMR (400MHz, CDCl3) 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 equivalent) in DCM (100 mL), TFA (15 mL) was added. The mixture was stirred at 20°C for 2 hours. The reaction mixture was then concentrated. The residue was diluted with  (150 mL), and aqueous NaHCO3 solution (150 mL) was added. Then CbzCl (4.0 g, 23.81 mmol, 1.2 equivalents) was added. The reaction mixture was stirred at 20°C for a further 1 hour. The product was extracted with  (50 mL x 3). The combined organic layers were washed with saline solution (50 mL), dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified by combi-flash ( in PE, 0%~30%) to obtain benzyl 3-acetyl-3-cyanopiperidine-1-carboxylate (293-3). 1 H NMR(400MHz,CDCl3)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 equivalent) and 2-((difluoromethyl)sulfonyl)pyridine (5 g, 26.19 mmol, 1.5 equivalents) in THF (50 mL), KHMDS (26 mL, 26.19 mmol, 1.5 equivalents) was added dropwise at -70°C under an N2 atmosphere. The reaction mixture was stirred at -70°C for 2 hours. The reaction product was quenched with aqueous NH4Cl solution (50 mL) and extracted with ELISA (50 mL x 3). The organic layer was washed with saline solution (100 mL), dried over anhydrous Na2SO4, filtered, and concentrated to obtain 3-cyano-3-(1,1-difluoro-2-hydroxy-1-(pyridine-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-(pyridine-2-ylsulfonyl)propan-2-yl)piperidine-1-carboxylate (293-4) (12 g, crude) in DMF (30 mL), saturated NH4Cl (30 mL) and aqueous HCl (30 mL, 6 M) were added. The reaction mixture was stirred at 80°C for 48 hours. The reaction mixture was cooled and diluted with H2O (50 mL). The mixture was extracted with RINKAN (50 mL x 3). The combined organic layers were washed with saline solution (100 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by combi-flash (RINKAN in PE, 0%~20%) to obtain benzyl 3-cyano-3-(1,1-difluoroprop-1-en-2-yl)piperidine-1-carboxylate (293-5). 1 H NMR(400MHz, CDCl3)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] 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 equivalent) in MeOH (100 mL) and 1 M NaOH solution (12.6 mL, 1.5 equivalents) was mixed with H2O2 (30%, 6.68 g, 59 mmol, 7.0 equivalents). The reaction mixture was stirred at 18°C ​​for 36 hours. The reaction mixture was quenched with saturated Na2S2O3 (50 mL). The aqueous layer was extracted with ELISA (50 mL x 3). The combined organic layers were dried over Na2SO4, filtered, concentrated, and dried. The residue was purified by combi-flash (0-40% siRNA in PE) to obtain benzyl 3-carbamoyl-3-(1,1-difluoroprop-1-en-2-yl)piperidine-1-carboxylate (293-6). 1H NMR (CDCl3400MHz): 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] 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 equivalent) and KOH (560 mg, 10 mmol, 2.5 equivalents) in MeOH (30 mL) was mixed with PhI(OAc)2 (1.53 g, 4.8 mmol, 1.2 equivalents) at 0°C. The resulting mixture was stirred at 15°C for 3 hours. The reaction product was quenched with saturated NH4Cl aqueous solution (50 mL). The aqueous layer was extracted with ELISA (50 mL x 3). The combined organic layers were dried over Na2SO4, filtered, and concentrated. The residue was purified by combi-flash (0-30% Â1 in PE) to obtain benzyl 3-(1,1-difluoroprop-1-en-2-yl)-3-((methoxycarbonyl)amino)piperidine-1-carboxylate (293-7). 1 H NMR(400MHz,CDCl3)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 equivalent) in DCM (15 mL), TMSI (977 mg, 4.89 mmol, 3.0 equivalent) was added. The reaction mixture was stirred at 16°C for 18 hours. The reaction mixture 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 freeze-dried to obtain 3-(1,1-difluoroprop-1-en-2-yl)piperidine-3-amine HCl salt (intermediate 293-8). 1 H NMR(400MHz,DMSO-d6)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)piperidine-3-amine [ka] To a solution of 1-benzyl 3-methyl 3-((tert-butoxycarbonyl)amino)piperidine-1,3-dicarboxylate (160 mg, 0.41 mmol) in MeOH (10 mL), LiCl (510 mg, 12.3 mmol) and NaBH4 (462 mg, 12.3 mmol) were 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 saline. The organic layer was dried over Na2SO4 and concentrated under vacuum to obtain crude alcohol. LC-MS: [M+H-100] + =265.0.

[0330] Dess-Martin reagent (782 mg, 1.844 mmol) was added to a solution of alcohol (560 mg, 1.537 mmol) in DCM (15 mL). The mixture was stirred at room temperature for 2 hours. The mixture was treated with aqueous NaHCO3 solution (10 mL) and the layers were separated. The aqueous layer was extracted with DCM (10 ml × 2). The combined organic layers were washed with saline solution, dried over Na2SO4, and concentrated. The filtrate was concentrated. The residue was purified and eluted with EA (0-50%) in n-hexane to obtain 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 mixed with an aqueous solution of CH3MgBr in 2-Me THF (3 M, 0.138 ml, 0.414 mmol) dropwise at -78°C. The mixture was stirred at -78°C for a further 1 hour and then warmed to 0°C for a further 2 hours. The mixture was quenched with an aqueous solution of NH4Cl (10 ml) and extracted with EA (20 ml x 3). The combined organic layers were washed with saline solution, dried over Na2SO4, and concentrated. The filtrate was concentrated. The residue was purified and eluted with EA (0-50%) in n-hexane to obtain alcohol. LC-MS: [M+H-100] + To a solution of alcohol (100 mg, 0.264 mmol) in 279.0.DCM (3 mL), TFA (0.5 mL) was added. The resulting mixture was stirred at room temperature for 2 hours. The mixture was concentrated under vacuum (below 40°C) to obtain 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), NsCl (657 mg, 2.96 mmol) was gradually added. The mixture was stirred overnight at room temperature under N2. The reaction product was treated with aqueous NaHCO3 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, dried over Na2SO4, and concentrated. The filtrate was concentrated. The residue was purified and eluted with n-hexane (0-30%) in EA to obtain benzyl 2-methyl-1-((4-nitrophenyl)sulfonyl)-1,5-diazaspiro[2.5]octane-5-carboxylate (295-3). LC-MS:[M+H] + =445.8.

[0333] 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) was mixed with 1 M TBAF (0.606 ml, 0.606 mmol) in THF under N2 conditions. The mixture was heated overnight to 45°C. The mixture was cooled to room temperature and treated with EA (15 mL) and H2O (10 mL). The layers were separated, and the aqueous layer was extracted with EA (15 mL x 3). The combined organic layers were washed with brine, dried over Na2SO4, and filtered. The filtrate was concentrated and purified by preparative HPLC to obtain benzyl 3-(1-fluoroethyl)-3-((4-nitrophenyl)sulfonamide)piperidine-1-carboxylate (295-4). LC-MS: [M+H] + =465.8.

[0334] A mixture of thioglycolic acid (71.2 mg, 0.773 mmol) and DBU235 (98 mg, 1.547 mmol) in CH3CN (10 mL) was stirred under N2 for 5 minutes. Then, benzyl 3-(1-fluoroethyl)-3-((4-nitrophenyl)sulfonamide)piperidine-1-carboxylate (295-4) (120 mg, 0.258 mmol) in CH3CN (2 mL) was added. The mixture was stirred overnight at room temperature and concentrated. The residue was treated with EA (10 mL), washed with aqueous NaHCO3 solution (5 mL x 2), H2O (10 mL), and brine, dried over Na2SO4, and filtered. The filtrate was concentrated, and the residue was purified by preparative HPLC to obtain benzyl 3-amino-3-(1-fluoroethyl)piperidine-1-carboxylate (295-5). LC-MS: [M+H] + =281.1.

[0335] A solution of benzyl 3-amino-3-(1-fluoroethyl)piperidine-1-carboxylate (295-5) (120 mg, 0.428 mmol) in MeOH (15 ml) was mixed with Pd(OH)2 (60 mg) under N2 conditions. The mixture was then stirred under an H2 balloon for 3 hours. The mixture was filtered, and the filtrate was concentrated to obtain 3-(1-fluoroethyl)piperidine-3-amine (intermediate 295-6). LC-MS: [M+H] + =147.1

[0336] Intermediate 297-6: 3-((methylthio)methyl)piperidine-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 equivalent) in THF (200 mL), MeSNa (11.0 g, 159 mmol, 4.0 equivalents) was added at 0°C. The resulting mixture was stirred at 15-20°C for 16 hours. The reaction product was quenched with water (300 mL) and extracted with ELISA (300 mL x 3). The combined organic layer was dried over anhydrous Na2SO4 and concentrated under vacuum. The residue was purified by Combi Flash (30% ELISA in PE) to obtain 1-benzyl 3-ethyl 3-((methylthio)methyl)piperidine-1,3-dicarboxylate (297-1). 1 H NMR(400MHz,CDCl3)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] 1-benzyl 3-ethyl 3-((methylthio)methyl)piperidine-1,3-dicarboxylate (297-1) (10.0 g, 28.5 mmol, 1.0 equivalent) and LiOH in MeOH / THF / H2O (120 mL, v / v / v=1 / 1 / 1) . A mixture of H2O (12.0 g, 285 mmol, 10.0 equivalents) was stirred at 20°C for 16 hours. The reaction was concentrated under vacuum. 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 ELISA (200 mL x 3). The combined organic layers were dried over anhydrous Na2SO4 and concentrated under vacuum to obtain 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 equivalent), DPPA (7.50 g, 27.2 mmol, 1.1 equivalent), and Et3N (5.00 g, 48.5 mmol, 2.0 equivalent) were stirred in toluene (100 mL) at 80°C for 1 hour under N2. The reaction mixture was then concentrated in vacuum. The residue was dissolved in dioxane (50 mL) and 6N HCl (50 mL), and the mixture was stirred at 25-30°C for 16 hours. The reaction mixture was then concentrated in vacuum. The pH of the aqueous layer was adjusted to 9 with 1N NaOH. The mixture was then extracted with ELISA (200 mL x 3). The combined organic layers were dried over anhydrous Na2SO4 and concentrated in vacuum. The residue was dissolved in DCM (100 mL), and DIPEA (10.0 g, 74.2 mmol, 3.0 equivalents) and Boc2O (11.0 g, 48.4 mmol) were added. The resulting mixture was stirred at 25-30°C for 16 hours. The reaction product was concentrated under vacuum. The residue was purified by Combi Flash (20% ELISA in PE) to obtain benzyl 3-((tert-butoxycarbonyl)amino)-3-((methylthio)methyl)piperidine-1-carboxylate (297-5). LC-MS: [M+H-100] + =295.1.

[0339] To a solution of benzyl 3-((tert-butoxycarbonyl)amino)-3-((methylthio)methyl)piperidine-1-carboxylate (297-5) (2.37 g, 6.00 mmol, 1 equivalent) in CH3CN (50 mL), TMSI (3.60 g, 18 mmol, 3 equivalents) was added dropwise at approximately 0°C. After addition, the reaction mixture was stirred at the same temperature for 2 hours. 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 x 4). The aqueous layer was freeze-dried to obtain crude 3-((methylthio)methyl)piperidine-3-amine (intermediate 297-6).

[0340] Intermediate 299-3: 2,2,2-trifluoroacetaldehyde compound and 1-(3-aminopiperidine-3-yl)-2,2,2-trifluoroethane-1-ol (1:1) [ka] A mixture of tert-butyl 3-nitropiperidine-1-carboxylate (5.0 g, 21.71 mmol, 1.0 equivalent) and 2,2,2-trifluoroethane-1,1-diol (10.08 g, 86.86 mmol, 4.0 equivalents) in THF (50 mL) was to be mixed with K2CO3 (1.8 g, 13.03 mmol, 0.6 equivalents) at 60°C for 16 hours. 1N HCl (10 mL) was added to the mixture and extracted with EA (40 mL x 3). The combined organic layer was washed with saline solution (30 mL), dried over Na2SO4, filtered, and concentrated under vacuum to obtain tert-butyl 3-nitro-3-(2,2,2-trifluoro-1-hydroxyethyl)piperidine-1-carboxylate (299-1). ¹H NMR: 1 H NMR (400MHz, CDCl3) 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] To a solution of tert-butyl 3-nitro-3-(2,2,2-trifluoro-1-hydroxyethyl)piperidine-1-carboxylate (299-1) (4.0 g, 12.89 mmol, 1.0 equivalent) and NH4Cl (3.45 g, 64.45 mmol, 5.0 equivalents) in EtOH / H2O (40 mL, v / v=4:1), Zn (8.43 g, 128.91 mmol, 10.0 equivalents) was added at 25°C under N2. The mixture was stirred at 80°C for 16 hours, work-up was performed under aqueous conditions, filtered, and concentrated under vacuum. The crude product was purified by column chromatography (PE:EA=10:1) to obtain tert-butyl 3-amino-3-(2,2,2-trifluoro-1-hydroxyethyl)piperidine-1-carboxylate (299-2). 1H NMR: 1 H NMR (400MHz, CDCl3) 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 at 25°C for 2 hours. The mixture was concentrated to obtain the 2,2,2-trifluoroacetaldehyde compound and 1-(3-aminopiperidine-3-yl)-2,2,2-trifluoroethane-1-ol (1:1) (intermediate 299-3).

[0343] Intermediate 300-6: (5-(3-amino-3-(1,2,2-trifluoroethyl)piperidine-1-yl)-2-(3,4-difluorophenyl)pyridine-4-yl)methanol [ka] To a solution of tert-butyl 3-nitropiperidine-1-carboxylate (3g, 13.03 mmol) in THF (30 mL), K2CO3 (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 stirred at 55°C under an N2 atmosphere for 18 hours. The reaction mixture was diluted with water and extracted with ELISA (20 mL x 3). The combined organic phase was washed with water (20 mL) and saline solution (20 mL), dried over anhydrous sodium sulfate, and concentrated under vacuum to obtain the residue. The residue was purified by flash chromatography (elution gradient: 0%~40% ELISA in PE, 30 minutes) to obtain 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 stirred at 60°C for 4 hours under an N2 atmosphere. The reaction mixture was diluted with water, extracted with ethyl acetate, and the combined organic phase was washed with water, saturated NaHCO3 solution, and saline solution. It was dried over sodium sulfate and concentrated to obtain the crude product. The reaction mixture was diluted with water, extracted with ethyl acetate (20 ml x 3), and the combined organic phase was washed with water (20 ml) and saline solution (20 ml). It was dried over anhydrous sodium sulfate and concentrated under vacuum to obtain the residue. The crude product was purified by flash chromatography (elution gradient: 0%~30% ethyl acetate in PE, 30 minutes) to obtain 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), Fe (2.432 g, 43.5 mmol) and NH4Cl (1.165 g, 21.77 mmol) were added, and the reaction mixture was stirred at 60°C for 18 hours under an N2 atmosphere. The reaction mixture was filtered, the filtrate was diluted with water, and extracted with RINKAN (30 ml x 3). The combined organic phase was washed with water (20 ml) and saline solution (20 ml), dried over anhydrous sodium sulfate, and concentrated under vacuum to obtain the residue. The crude product was purified by flash chromatography (elution gradient: 0%~50% RINKAN in PE, 30 minutes) to obtain 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), TFA (4 mL) was added, and the reaction mixture was stirred at room temperature under an N2 atmosphere for 1 hour. The reaction mixture was concentrated to obtain 3-(1,2,2-trifluoroethyl)piperidine-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)piperidine-3-amine (300-4) (1.3 g, 3.17 mmol) in DMSO (30 mL), DIPEA (15 mL) was added at room temperature, and the reaction mixture was stirred at 80 °C for 8 hours under an N2 atmosphere. The reaction mixture was diluted with water, extracted with ELISA (20 ml x 3), and a workup was performed under aqueous conditions. The mixture was concentrated under vacuum to obtain the residue. The crude product was purified by flash chromatography (elution gradient: 0%~30% ELISA in PE, 30 minutes) to obtain methyl 5-(3-amino-3-(1,2,2-trifluoroethyl)piperidine-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)piperidine-1-yl)-2-(3,4-difluorophenyl)isonicotinate 300-5) (0.65 g, 1.514 mmol) in THF (20 mL) and methanol (20 mL), lithium chloride (1.284 g, 30.3 mmol) and sodium tetrahydroborate (1.145 g, 30.3 mmol) were added at room temperature, and the reaction mixture was stirred at room temperature under an N2 atmosphere for 3 hours. The reaction mixture was diluted with water and extracted with ELISA (20 mL x 3). The combined organic phase was washed with water (20 mL) and saline solution (20 mL), dried over anhydrous sodium sulfate, and concentrated under vacuum to obtain the residue. The crude product was purified by flash chromatography (elution gradient: 10%-40% ethyl acetate in PE, 30 minutes) to obtain (5-(3-amino-3-(1,2,2-trifluoroethyl)piperidine-1-yl)-2-(3,4-difluorophenyl)pyridine-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)piperidine-1-yl)-2-(3,4-difluorophenyl)pyridine-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), DEAD (0.592 mL, 3.74 mmol) was added dropwise at 0°C, and the reaction mixture was stirred at 0°C for 0.5 hours under an N2 atmosphere. The reaction mixture was diluted with water and extracted with ELISA (20 ml x 3). The combined organic phase was washed with water (20 ml) and saline solution (20 ml), dried over anhydrous sodium sulfate, and concentrated under vacuum to obtain the residue. The crude product was purified by flash chromatography (elution gradient: 0%~10% MeOH in DCM, 50 minutes) to obtain intermediates 300-7. LC-MS: [M+H] + =719.2.

[0350] Intermediate 301-8: 3-amino-1-(4-(chloromethyl)-6-(3,4-difluorophenyl)pyridine-3-yl)-3-(2,2-difluoroethyl)piperidine-4-ol [ka] To a solution of LDA (6.94 mL, 13.88 mmol) in THF (9 mL), a solution of 1-(tert-butyl)3-methyl4-hydroxypiperidine-1,3-dicarboxylate (1.5 g, 5.78 mmol) in THF was added at -78°C under an N2 atmosphere. The mixture was stirred at this temperature for 10 minutes. Then, 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 stirred at room temperature under an N2 atmosphere for 30 minutes. The reaction mixture was diluted with water and extracted with ELISA (20 ml x 3). The combined organic phase was washed with water (20 ml) and saline solution (20 ml), dried over anhydrous sodium sulfate, and concentrated under vacuum to obtain the residue. The residue was purified by flash chromatography (elution gradient: 0%~30% phenylethylamine in PE, 30 minutes) to obtain 1-(tert-butyl)3-methyl3-(2,2-difluoroethyl)-4-hydroxypiperidine-1,3-dicarboxylate (301-1). LC-MS: [M+H] + =324.1.

[0351] To a solution of 1-(tert-butyl)3-methyl3-(2,2-difluoroethyl)-4-hydroxypiperidine-1,3-dicarboxylate (301-1) (0.6 g, 1.856 mmol) in methanol (5 mL), a solution of LiOH (700 mg, 29.2 mmol) in water (5 mL) was added at room temperature under an N2 atmosphere, and the reaction mixture was stirred at room temperature for 5 hours. The pH of the reaction mixture was adjusted to 4 by adding a 3 M HCl solution, extracted with ELISA (20 ml x 3), washed with water (20 ml) and saline solution (20 ml), dried over anhydrous sodium sulfate, and concentrated under vacuum to obtain 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 at room temperature under an N2 atmosphere, and the reaction mixture was stirred at 100 °C for 18 hours. The reaction mixture was diluted with water, extracted with ELISA (20 ml x 3), and the combined organic phase was washed with water (20 ml) and saline solution (20 ml). The mixture was dried over anhydrous sodium sulfate and concentrated under vacuum to obtain the residue. The residue was purified by flash chromatography (elution gradient: 0%~50% ethyl acetate in PE, 30 minutes) to obtain tert-butyl 3a-(2,2-difluoroethyl)-2-oxohexahydrooxazolo[4,5-c]pyridine-5(4H)-carboxylate (303-3). 1 H NMR(400MHz,DMSO-d6)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), TFA (3 mL, 38.9 mmol) was added at room temperature under an N2 atmosphere, and the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated under vacuum to obtain crude 3a-(2,2-difluoroethyl)hexahydrooxazolo[4,5-c]pyridine-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]pyridine-2(3H)-one (301-4) in DMSO (10 mL), DIPEA (10 mL, 57.3 mmol) was added at room temperature under an N2 atmosphere, and the reaction mixture was stirred at 120 °C for 18 hours. The reaction mixture was diluted with water, extracted with ELISA (20 ml x 3), washed with water (20 ml x 3) and saline solution (20 ml), dried over anhydrous sodium sulfate, concentrated under vacuum, and the residue was obtained. The residue was purified by flash chromatography (elution gradient: 0%~10% MeOH in DCM, 30 minutes) to obtain methyl 5-(3a-(2,2-difluoroethyl)-2-oxohexahydrooxazolo[4,5-c]pyridine-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]pyridine-5(4H)-yl)-2-(3,4-difluorophenyl)isonicotinate (301-5) (40 mg, 0.088 mmol) in THF (5 mL) and methanol (5 mL), LiCl (74.8 mg, 1.764 mmol) and NaBH4 (33.4 mg, 0.882 mmol) were added at room temperature under an N2 atmosphere, and the reaction mixture was stirred at room temperature for 5 hours. The reaction mixture was diluted with water, extracted with RINKAN (20 ml x 3), washed with water (20 ml) and saline solution (20 ml), dried over anhydrous sodium sulfate, and concentrated under vacuum to obtain the residue. The residue was purified by flash chromatography (elution gradient: 0%~10% MeOH in DCM, 30 minutes) to obtain 3a-(2,2-difluoroethyl)-5-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridine-3-yl)hexahydrooxazolo[4,5-c]pyridine-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)pyridine-3-yl)hexahydrooxazolo[4,5-c]pyridine-2(3H)-one (301-6) (100 mg, 0.235 mmol) in MeOH (10 mL) and water (10 mL), KOH (132 mg, 2.351 mmol) was added at room temperature under an N2 atmosphere, and the reaction mixture was stirred at 100 °C for 18 hours. The reaction mixture was diluted with water, extracted with ELISA (20 ml x 3), washed with water (20 ml) and saline solution (20 ml), dried over anhydrous sodium sulfate, and concentrated under vacuum to obtain the residue. The residue was purified by flash chromatography (elution gradient of 0% to 10% MeOH in DCM for 30 minutes) to obtain 3-amino-3-(2,2-difluoroethyl)-1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridine-3-yl)piperidine-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)pyridine-3-yl)piperidine-4-ol (301-7) (130 mg, 0.325 mmol) in DCM (10 mL), thionyl chloride (0.119 mL, 1.627 mmol) was added at 0°C under an N2 atmosphere, and the reaction mixture was stirred at room temperature for 5 hours. The reaction mixture was quenched at 0°C with 5 mL of saturated K2CO3 aqueous solution. The reaction mixture was extracted with DCM (20 mL x 3), the combined organic phase was washed with water (20 mL) and saline solution (20 mL), dried over anhydrous sodium sulfate, and concentrated under vacuum to obtain crude 3-amino-1-(4-(chloromethyl)-6-(3,4-difluorophenyl)pyridine-3-yl)-3-(2,2-difluoroethyl)piperidine-4-ol (intermediate 301-8). LC-MS: [M+H] + =418.1.

[0358] Intermediate 302-9: 3-amino-3-(6-chloropyridine-2-yl)-1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridine-3-yl)piperidine-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), TfOH (22.25 mL, 251 mmol) was added dropwise at 0°C. The reaction mixture was then stirred at room temperature for 2 hours, and mesitylene (9.54 mL, 68.9 mmol) was added dropwise at 0°C. The reaction mixture was stirred under an N2 atmosphere at room temperature for 18 hours. The solvent was removed under vacuum to obtain the crude product. The crude product was recrystallized in Et2O at -20°C to obtain (6-chloropyridine-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. The reaction mixture was stirred at room temperature for 10 minutes, then (6-chloropyridine-2-yl)(mesityl)iodonium trifluoromethanesulfonate (intermediate 302-1) (3.52 g, 5.56 mmol) was added at room temperature. The reaction mixture was stirred at room temperature under an N2 atmosphere for 18 hours. The reaction mixture was diluted with water, extracted with ethyl acetate, washed with water and saline solution, dried over sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by flash chromatography (elution gradient: 0%~30% EtOAC in hexane, 30 minutes) to obtain 1-(tert-butyl)3-methyl3-(6-chloropyridine-2-yl)-4-oxopiperidine-1,3-dicarboxylate (302-2). LC-MS: [M+H] + =369.0

[0360] To a solution of 1-(tert-butyl)3-methyl3-(6-chloropyridine-2-yl)-4-oxopiperidine-1,3-dicarboxylate (intermediate 302-2) (1.0 g, 2.71 mmol) in MeOH (30 mL), sodium tetrahydroborate (0.205 g, 5.42 mmol) was added at 0°C, and the reaction mixture was stirred at 0°C for 30 minutes. The reaction mixture was diluted with water, extracted with ELISA, washed with water and brine, dried over sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by flash chromatography (elution gradient: 0%~30% EtOAC in hexane, 30 minutes) to obtain 1-(tert-butyl)3-methyl3-(6-chloropyridine-2-yl)-4-hydroxypiperidine-1,3-dicarboxylate (302-3). LC-MS: [M+H] + =371.0

[0361] To a solution of 1-(tert-butyl)3-methyl3-(6-chloropyridine-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 stirred at room temperature under an N2 atmosphere for 5 hours. The pH of the reaction mixture was adjusted to 4 by adding 5 M aqueous HCl, extracted with ELISA, washed with water and brine, dried over sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by flash chromatography (elution gradient: 0%~10% MeOH in DCM, 30 minutes) to obtain 1-(tert-butoxycarbonyl)-3-(6-chloropyridine-2-yl)-4-hydroxypiperidine-3-carboxylate (302-4). LC-MS: [M+H] + =357.1.

[0362] To a solution of 1-(tert-butoxycarbonyl)-3-(6-chloropyridine-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. The reaction mixture was stirred at 100°C for 3 hours under an N2 atmosphere, then methanol (0.091 mL, 2.242 mmol) was added, and the reaction mixture was stirred at 100°C for 18 hours under an N2 atmosphere. The reaction mixture was diluted with water, extracted with siRNA, washed with water and brine, dried over sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by flash chromatography (elution gradient: 0%~10% MeOH in DCM, 30 minutes) to obtain tert-butyl3a-(6-chloropyridine-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-chloropyridine-2-yl)-2-oxohexahydrooxazolo[4,5-c]pyridine-5(4H)-carboxylate (302-5) (400 mg, 0.113 mmol) in DCM (9 mL), TFA (3 mL) was added at room temperature, and the reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was concentrated to obtain crude 3a-(6-chloropyridine-2-yl)hexahydrooxazolo[4,5-c]pyridine-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-chloropyridine-2-yl)hexahydrooxazolo[4,5-c]pyridine-2(3H)-one (302-6) (120 mg, 0.326 mmol) in DMSO (5 mL), DIPEA (5 mL) was added, and the reaction mixture was stirred at 100 °C for 5 hours under an N2 atmosphere. The reaction mixture was diluted with water, extracted with ethyl acetate, washed with water and saline solution, dried over sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by flash chromatography (elution gradient: 0%~10% MeOH in DCM, 30 minutes) to obtain methyl 5-(3a-(6-chloropyridine-2-yl)-2-oxohexahydrooxazolo[4,5-c]pyridine-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-chloropyridine-2-yl)-2-oxohexahydrooxazolo[4,5-c]pyridine-5(4H)-yl)-2-(3,4-difluorophenyl)isonicotinate (302-7) (70 mg, 0.140 mmol) in methanol (5 mL) and THF (5 mL), lithium chloride (59.2 mg, 1.398 mmol) and sodium tetrahydroborate (52.9 mg, 1.398 mmol) were added, and the reaction mixture was stirred at room temperature under an N2 atmosphere for 5 hours. The reaction mixture was diluted with water, extracted with ethyl acetate, washed with water and saline solution, dried over sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by flash chromatography (elution gradient: 0%~10% MeOH in DCM, 30 minutes) to obtain 3a-(6-chloropyridine-2-yl)-5-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridine-3-yl)hexahydrooxazolo[4,5-c]pyridine-2(3H)-one(302-8). LC-MS:[M+H] + =473.1.

[0366] To a solution of 3a-(6-chloropyridine-2-yl)-5-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridine-3-yl)hexahydrooxazolo[4,5-c]pyridine-2(3H)-one (302-8) (50 mg, 0.106 mmol) in methanol (5 mL), a solution of KOH (59.3 mg, 1.057 mmol) in water (5 mL) was added at room temperature, and the reaction mixture was stirred at 100 °C for 8 hours under an N2 atmosphere. The reaction mixture was diluted with water, extracted with ethyl acetate, washed with water and saline solution, dried over sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by flash chromatography (elution gradient: 0%~10% MeOH in DCM, 30 minutes) to obtain 3-amino-3-(6-chloropyridine-2-yl)-1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridine-3-yl)piperidine-4-ol (intermediate 302-9). LC-MS: [M+H] + =447.1

[0367] Intermediate 303-3: 3-(2,2,2-trifluoroethyl)piperidine-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 DMA (50 mL) under N2. Then, reagent 1 (3 g, 12.97 mmol) and TMSN3 (2.99 g, 25.9 mmol) were added. The mixture was stirred overnight at room temperature. The mixture was treated with EA (50 mL) and H2O (50 mL). The layers were separated, and the aqueous layer was extracted with EA (30 ml × 3). The combined organic layers were washed with saline solution (20 ml), dried over Na2SO4, and filtered. The filtrate was concentrated to purify the residue, which was eluted with EA in n-hexane (0-20%) to obtain benzyl 3-azido-3-(2,2,2-trifluoroethyl)piperidine-1-carboxylate (303-1). 1H NMR(400MHz, CDCl37.31-7.51(m,1H),5.03-5.34(m,1H),3.86(d,J=12.80Hz,1H),3.20(d,J=1 3.55Hz,1H),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 mixed with indium powder (1.3 g, 11.4 mmol) and NH4Cl (0.609 g, 11.38 mmol). The mixture was heated under reflux for 2 hours in a sealed tube. The mixture was cooled to room temperature and filtered. The filtrate was concentrated. The residue was purified by preparative HPLC to obtain 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) was added under N2 conditions. The mixture was then stirred under an H2 balloon for 3 hours. The mixture was filtered, and the filtrate was concentrated to obtain 3-(2,2,2-trifluoroethyl)piperidine-3-amine (intermediate 303-3). 1 H NMR(CDCl3)δ:3.57-3.35(m,4H),3.02-2.84(m,2H),2.43(q,J=11.1Hz,2H),2.25(dddd ,J=17.3,13.2,8.6,4.2Hz,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)pyridine-3-yl)-3-ethynylpiperidine-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 equivalent) and dimethyl (1-diazo-2-oxopropyl)phosphonate (7.8 g, 40.8 mmol, 2.0 equivalents) in dry methanol (120 mL), K2CO3 (8.5 g, 61.2 mmol, 3.0 equivalents) was added. The mixture was stirred at 1-7°C for 18 hours, filtered, and the filtrate was concentrated. The residue was purified by flash chromatography (PE / EA = 70 / 30) to obtain 1-benzyl 3-methyl 3-ethynylpiperidine-1,3-dicarboxylate (304-1). 1 H NMR(CDCl3400MHz):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 equivalent) in THF (50 mL) and water (15 mL), LiOH.H2O (1.7 g, 41.1 mmol, 4.0 equivalents) was added. The mixture was stirred at 50°C for 2 hours. 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 (aqueous solution). The suspension was extracted with DCM (80 mL x 2), dried over Na2SO4, and concentrated to obtain crude 1-((benzyloxy)carbonyl)-3-ethynylpiperidine-3-carboxylate (304-2). 1H NMR(CDCl3400MHz):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 equivalent) in DCM (125 mL), NH4Cl (1 g, 18.8 mmol, 2.0 equivalents) was added. HATU (7.1 g, 18.8 mmol, 2.0 equivalents) and DIEA (4.9 g, 37.6 mmol, 4 equivalents) were added. The reaction mixture was stirred at 25°C for 16 hours. The mixture was washed with water (100 mL) and saline solution (150 mL), dried over Na2SO4, and concentrated. The residue was purified by flash chromatography (PE / EA = 50 / 50) to obtain benzyl 3-carbamoyl-3-ethynylpiperidine-1-carboxylate (304-3). 1 H NMR(400MHz,CDCl3):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 equivalent) in methanol (100 mL), KOH (1.2 g, 21.8 mmol, 2.5 equivalents) was added. Then, PhI(OAc)2 (2.8 g, 8.7 mmol, 1.0 equivalent) was added. The resulting mixture was stirred at 3-10°C for 30 minutes. The color changed from colorless to yellow. The reaction product was concentrated, diluted with water (150 mL), extracted with EA (100 mL x 2), dried over Na2SO4, and concentrated. The residue was purified by flash chromatography (PE / EA = 70 / 30) to obtain benzyl 3-ethynyl-3-((methoxycarbonyl)amino)piperidine-1-carboxylate (304-4). 1 H NMR(400MHz,CD3OD):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 equivalent) and TFA (10 mL) in DCM (10 mL) was stirred at 75°C for 12 hours. The mixture was concentrated, diluted with DCM (20 mL), poured into saturated NaHCO3 solution (50 mL), extracted with CHCl3 / i-PrOH=3 / 1 (50 mL × 4), dried over Na2SO4, and concentrated to obtain crude methyl (3-ethynylpiperidine-3-yl)carbamate (304-5). LCMS:[M+H] + =182.9.

[0375] A mixture of methyl (3-ethynylpiperidine-3-yl)carbamate (304-5) (600 mg, 2.53 mmol, 1.0 equivalent) and methyl 2-(3,4-difluorophenyl)-5-fluoroisonicotinate (intermediate A) (670 mg, 2.53 mmol, 1.0 equivalent) in DMSO (10 mL) was mixed with DIEA (2.3 g, 7.0 equivalents). The reaction mixture was stirred at 120 °C for 2.5 hours. The mixture was cooled, diluted with water (100 mL), extracted with EA (50 mL x 2), washed with saline solution (100 mL), dried over Na2SO4, and concentrated. The residue was purified by flash chromatography (PE / EA = 10 / 1 to 70 / 30) to obtain methyl 2-(3,4-difluorophenyl)-5-(3-ethynyl-3-((methoxycarbonyl)amino)piperidine-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)piperidine-1-yl) isonicotinate (intermediate 304-6) (280 mg, 0.652 mmol, 1.0 equivalent) in dry THF (12 mL), L-selectlide (3.9 mL) was added dropwise at 0°C under N2. The solution was stirred at 0°C for 30 minutes. The mixture was quenched with saturated NH4Cl solution (30 mL), diluted with water (100 mL), extracted with EA (50 mL x 2), dried over Na2SO4, and concentrated. The residue was purified by flash chromatography (PE / EA=1 / 1) to obtain methyl(1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridine-3-yl)-3-ethynylpiperidine-3-yl)carbamate (intermediate 304-7). LCMS: 402.1[M+H] + =402.1.

[0377] Intermediate 305-5: 3-(difluoromethyl)piperidine-3-amine [ka] To a solution of 1-benzyl 3-ethyl 3-formylpiperidine-1,3-dicarboxylate (7.0 g, 21.9 mmol, 1.0 equivalent) in 100 mL of DCM at 0°C under an N2 atmosphere, DAST (14.1 g, 87.8 mmol, 4.0 equivalents) was added dropwise. The reaction mixture was warmed to 10°C and stirred for 16 hours. The reaction mixture was cooled to 0°C and carefully quenched with 100 mL of NaHCO3 aqueous solution. The mixture was extracted with 3 x 50 mL of DCM. The combined organic layers were dried over Na2SO4, filtered, and concentrated under vacuum. The residue was purified using a silica gel column (PE / EA = 10 / 1) to obtain 4.0 g of crude product, which was further purified by preparative HPLC (with 0.1% TFA as an additive) to obtain 1-benzyl 3-ethyl 3-(difluoromethyl)piperidine-1,3-dicarboxylate 1-benzyl(305-1). 1 H NMR(400MHz,CDCl3):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] A mixture of 1-benzyl 3-ethyl 3-(difluoromethyl)piperidine-1,3-dicarboxylate (305-1) (2.6 g, 7.62 mmol, 1 equivalent) in THF (10 mL), MeOH (10 mL), and H2O (10 mL) was mixed with LiOH.H2O (3.2 g, 76.2 mmol, 10 equivalents). The reaction mixture was stirred at 40°C for 16 hours. The reaction mixture was cooled, acidified to pH=3 with 6 M HCl, and then extracted with DCM (30 mL x 3). The combined organic layers were washed with saline solution (50 mL), dried over Na2SO4, filtered, and concentrated to obtain 1-((benzyloxy)carbonyl)-3-(difluoromethyl)piperidine-3-carboxylate (305-2). 1H NMR(400MHz,CDCl3):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] A mixture of 1-((benzyloxy)carbonyl)-3-(difluoromethyl)piperidine-3-carboxylic acid (305-2) (3.4 g, 10.9 mmol, 1.0 equivalent) and Et3N (2.2 g, 21.72 mmol, 2.0 equivalent) in toluene (50 mL) under an N2 atmosphere was mixed with DPPA (3.3 g, 11.9 mmol, 1.1 equivalent). The reaction mixture was heated and stirred at 60°C for 3 hours. The reaction mixture was cooled, diluted with ELISA (100 mL), and then washed with H2O (50 mL x 2). The organic layer was dried over Na2SO4, filtered, and concentrated to obtain 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 equivalent) in dioxane (10 mL), 6 M HCl (10 mL) was added. The reaction mixture was stirred at 40 °C for 16 hours. The reaction mixture was concentrated to remove the dioxane, then diluted with H₂O (20 mL) and extracted with ELISA (20 mL). The aqueous phase was basicized to pH=10 with aqueous NaOH solution. The mixture was extracted with DCM (50 mL × 3). The combined organic layers were dried over Na₂SO₄, filtered, and concentrated to obtain benzyl 3-amino-3-(difluoromethyl)piperidine-1-carboxylate (305-4). 1H NMR(400MHz,CDCl3):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 equivalent) and Pd / C (10%, 1.5 g) in EtOH (36 mL) was hydrogenated under an H2 balloon for 3 hours. The reaction mixture was filtered, and the filtrate was concentrated and dried to obtain 3-(difluoromethyl)piperidine-3-amine (intermediate 305-5). 1 H NMR(400MHz,CD3OD):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)piperidine-1-yl)-2-(3-fluoro-4-methoxyphenyl)pyridine-4-yl)methyl)-9H-purine-6-yl)carbamate [ka] A mixture of ethyl 5-bromonicotinate (20.0 g, 86.9 mmol, 1 equivalent), NH2Boc (12.2 g, 104 mmol, 1.2 equivalents), Xant-Phos (1.60 g, 2.77 mmol, 0.03 equivalents), Pd2(dba)3 (1.60 g, 1.75 mmol, 0.02 equivalents), and Cs2CO3 (40.0 g, 123 mmol, 1.4 equivalents), along with dioxane (300 mL), was stirred at 110°C for 16 hours under an N2 atmosphere. The reaction mixture was then concentrated and dried. Water (800 mL) was added, and the reaction mixture was extracted with siRNA (300 mL x 2). The combined organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by combi flash (siRNA / PE = 0-40%) to obtain ethyl 5-((tert-butoxycarbonyl)amino)nicotinate (306-1). 1 H NMR(400MHz,CDCl3):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] A solution of 3.05 g, 80.3 mmol, 1.6 equivalents of LAH in 200 mL of THF was mixed dropwise with a solution of ethyl 5-((tert-butoxycarbonyl)amino)nicotinate (306-1) (13.5 g, 50.7 mmol, 1 equivalent) in 200 mL of THF at 0°C. After addition, the reaction mixture was stirred at approximately 0°C for 2 hours. 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 and dried. The residue was purified by combi Flash (Â / PE = 0-100%) to obtain tert-butyl(5-(hydroxymethyl)pyridine-3-yl)carbamate (306-2). 1 HNMR (400MHz, CDCl3): δ 8.33 (d, J = 2.4 Hz, 1H), 8.22 (d, J = 1.6 Hz, 1H), 8.01 (s, 1H), 7.23 (brs, 1H), 4.69 (s, 2H), 1.53 (s, 9H).

[0384] A mixture of tert-butyl(5-(hydroxymethyl)pyridine-3-yl)carbamate (intermediate 306-2) (10.0 g, 44.6 mmol, 1 equivalent) and PtO2 (3.0 g) in AcOH (3 mL) and EtOH (300 mL) was hydrogenated at 50°C for 7 days under an H2 atmosphere (50 psi). The reaction mixture was filtered, the filtrate was concentrated and dried to obtain tert-butyl(5-(hydroxymethyl)piperidine-3-yl)carbamate (306-3). LC-MS: [M+H] + =230.1

[0385] To a suspension of tert-butyl(5-(hydroxymethyl)piperidine-3-yl)carbamate (306-3) (10.0 g, impurity, 1 equivalent) and NaHCO3 (10.9 g, 130 mmol, 3 equivalents) in THF (100 mL) and H2O (100 mL), CbzCl (8.20 g, 47.4 mmol, 1.1 equivalents) was added dropwise at approximately 0°C. After addition, the reaction mixture was stirred at approximately 20°C for 16 hours. The reaction mixture was poured into water (400 mL) and extracted with ethyl acetate (200 mL x 3). The combined organic layer was dried over Na2SO4, concentrated, and dried. The residue was purified by combi flash (ethyl acetate / PE = 0-70%) to obtain benzyl 3-((tert-butoxycarbonyl)amino)-5-(hydroxymethyl)piperidine-1-carboxylate benzyl (306-4). 1 H NMR(400MHz,CDCl3):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] A mixture of benzyl 3-((tert-butoxycarbonyl)amino)-5-(hydroxymethyl)piperidine-1-carboxylate (306-4) (3.50 g, 9.60 mmol, 1.0 equivalent) in THF (60 mL) was mixed with Et3N (13.6 g, 134 mmol, 14.0 equivalents), FSO2(CF2)3CF3 (5.80 g, 19.2 mmol, 2.0 equivalents), and TEA.3HF (6.19 g, 38.4 mmol, 4.0 equivalents). The reaction mixture was stirred at 25°C for 12 hours. The reaction mixture was concentrated under vacuum. The residue was purified by silica gel column chromatography (eluate: PE / EA10 / 1~3 / 1) to obtain benzyl 3-((tert-butoxycarbonyl)amino)-5-(fluoromethyl)piperidine-1-carboxylate (306-5). LC-MS:[M+Na] + =389.1.

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

[0388] To a solution of tert-butyl(5-(fluoromethyl)piperidine-3-yl)carbamate (306-6) (1.50 g, 6.51 mmol, 1.0 equivalent) in DMSO (15 mL), DIEA (5.89 g, 45.6 mmol, 7.0 equivalents) and methyl 5-fluoro-2-(3-fluoro-4-methoxyphenyl) isonicotinate (intermediate D) (1.82 g, 6.51 mmol, 1.0 equivalent) were added. The reaction mixture was stirred at 120 °C for 3 hours under N2. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with saline solution (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography (PE / EA10 / 1~2 / 1), and further purified by preparative HPLC (with 0.05% NH3·H2O as an additive) to obtain methyl 5-((3R,5R)-3-((tert-butoxycarbonyl)amino)-5-(fluoromethyl)piperidine-1-yl)-2-(3-fluoro-4-methoxyphenyl)isonicotinate (306-7). 1 H NMR(400MHz,CD3OD):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)piperidine-1-yl)-2-(3-fluoro-4-methoxyphenyl)isonicotinate (306-7) (120 mg, 0.24 mmol, 1.0 equivalent) and LiCl (102 mg, 2.44 mmol, 10 equivalents) in THF (2 mL) and MeOH (2 mL), NaBH4 (181 mg, 4.88 mmol, 20 equivalents) was gradually added. After the addition, the mixture was stirred at 25°C for 1 hour. Then, water (20 mL) was added, and the mixture was extracted with ELISA (10 mL x 3). The combined organic phases were washed with saline solution (10 mL x 2), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum to obtain tert-butyl((3R,5R)-1-(6-(3-fluoro-4-methoxyphenyl)-4-(hydroxymethyl)pyridine-3-yl)-5-(fluoromethyl)piperidine-3-yl)carbamate (306-8). LC-MS:[M+H] + =464.3

[0390] DIAD (66.0 mg, 0.32 mmol, 1.5 equivalents) was added dropwise to a mixture of tert-butyl((3R,5R)-1-(6-(3-fluoro-4-methoxyphenyl)-4-(hydroxymethyl)pyridine-3-yl)-5-(fluoromethyl)piperidine-3-yl) carbamate (intermediate 306-8) (100 mg, 0.22 mmol, 1.0 equivalent, intermediate B (109 mg, 0.32 mmol, 1.5 equivalents), n-Bu3P (66.0 mg, 0.32 mmol, 1.5 equivalents), and THF (10 mL)). After addition, the mixture was stirred under N2 at 25°C for 12 hours. The mixture was concentrated under vacuum. The residue was purified by silica gel column chromatography (PE / EA = 5:1 to 1:1) to obtain tert-butyl(tert-butoxycarbonyl(9-((5-((3R,5R)-3-(tert-butoxycarbonyl)amino)-5-(fluoromethyl)piperidine-1-yl)-2-(3-fluoro-4-methoxyphenyl)pyridine-4-yl)methyl)-9H-purine-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)piperidine-1-yl)-2-(3,4-difluorophenyl)isonicotinate [ka] To a solution of 3-aminopicolinic acid (50 g, 360 mmol, 1.0 equivalent) in anhydrous MeOH (500 mL), concentrated H2SO4 (106 g, 1090 mmol, 3.0 equivalents) was added dropwise. The reaction mixture was stirred at 80°C for 48 hours. SOCl2 (130 g, 1090 mmol, 3.0 equivalents) was added to the reaction mixture, and the mixture was stirred at 80°C for another 48 hours. The reaction mixture was concentrated, diluted with water (500 mL), basicized to pH=9 with K2CO3(s), extracted with EA (500 mL x 3), washed with saline solution (500 mL), dried over Na2SO4, and concentrated to obtain methyl 3-aminopicolinate (311-1). 1 H NMR (400MHz, CDCl3) 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] To a solution of methyl 3-aminopicolinate (311-1) (27 g, 180 mmol, 1.0 equivalent) and K2CO3 (48 g, 350 mmol, 2.0 equivalents) in MeCN (500 mL), methyl carbonochloride (33 g, 350 mmol, 2.0 equivalents) was added at 20°C for 18 hours. The reaction mixture was diluted with water (500 mL) and concentrated. The residue was acidified to pH=3 with 1N HCl, extracted with EA (500 mL x 3), washed with saturated NaHCO3 (500 mL) and saline solution (300 mL), dried over Na2SO4, and concentrated to obtain methyl 3-((methoxycarbonyl)amino)picolinate (311-2). 1H NMR(400MHz, CDCl3)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] A solution of methyl 3-((methoxycarbonyl)amino)picolinate (intermediate 311-2) (22 g, 105 mmol, 1.0 equivalent) and PtO2 (5 g, 23 wt%) in AcOH (220 mL) was stirred under H2 at 25°C and 5 MPa for 144 hours. The reaction mixture was filtered and concentrated to obtain 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, approximately 105 mmol, 1.0 equivalent) in THF (300 mL), TEA (53 g, 525 mmol, 5.0 equivalent) and Boc2O (69 g, 315 mmol, 3.0 equivalent) were added at 20°C for 18 hours. The reaction mixture was concentrated, diluted with water (100 mL), acidified to pH=3 with 1N HCl, extracted with EA (100 mL x 3), washed with saline solution (100 mL), dried over Na2SO4, and concentrated. The residue was purified by combi-flash (EA%=0%~50%) to obtain 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 equivalent) in anhydrous THF (660 mL), LiAlH4 (5.15 g, 136 mmol, 1.3 equivalents) was added at -78°C to -10°C for 2 hours. The reaction mixture was poured into saturated NH4Cl (700 mL), acidified to pH=5 with 1N HCl, extracted with EA (500 mL x 3), washed with saline solution (500 mL), dried over Na2SO4, and concentrated. The residue was purified by combi-flash (EA%=0%~70%) to obtain tert-butyl(2S,3R)-2-(hydroxymethyl)-3-((methoxycarbonyl)amino)piperidine-1-carboxylate (311-5). 1 H NMR(400MHz,CDCl3)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] A mixture of tert-butyl(2S,3R)-2-(hydroxymethyl)-3-((methoxycarbonyl)amino)piperidine-1-carboxylate (311-5) (5g, 17 mmol, 1.0 equivalent), BnBr (12g, 69 mmol, 1.0 equivalent), and TBAI (3.1g, 9 mmol, 0.5 equivalents) in anhydrous THF (50mL) was to be mixed with NaH (2.6g, 65 mmol, 3.8 equivalents) at 20°C. After the addition, the reaction mixture was stirred at 60°C for 2 hours. The reaction mixture was poured into a solution of saturated NH4Cl (200mL), extracted with EA (100mL x 3), washed with saline solution (50mL), dried over Na2SO4, and concentrated. The residue was purified by combi-flash (EA%=0%~50%) to obtain tert-butyl(2S,3R)-3-(benzyl(methoxycarbonyl)amino)-2-((benzyloxy)methyl)piperidine-1-carboxylate (311-6). 1 H NMR(400MHz,CDCl3)δ:7.53-6.90(m,10H),4.90-3.31(m,12H),3.08-2.74(m,1 H),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 equivalent) in DCM (40 mL) and TFA (20 mL) was stirred at 20°C for 20 hours. The reaction mixture was diluted with H2O (100 mL), basicized to pH=9 with K2CO3(s), extracted with EA (100 mL x 3), washed with saline, dried over Na2SO4, and concentrated to obtain methylbenzyl((2S,3R)-2-((benzyloxy)methyl)piperidine-3-yl)carbamate (311-7). LC-MS:[M+H] + =369.5.

[0398] A mixture of methyl 2-(3,4-difluorophenyl)-5-fluoroisonicotinate (intermediate A) (30 g, 0.112 mmol, 1.0 equivalent) and m-CPBA (30 g, 0.174 mmol, 1.55 equivalents) in CDCl3 (500 mL) was stirred at 65°C for 20 hours. Further m-CPBA (30 g, 0.174 mmol, 1.55 equivalents) was added to the reaction mixture, and the reaction was stirred at 65°C for 72 hours. The reaction mixture was diluted with water (300 mL), basicized to pH=8 with K2CO3(s), extracted with EA (300 mL x 3), washed with saline solution (300 mL), dried over Na2SO4, and concentrated. The crude product was purified by grinding with PE / MTBE = 1 / 1 (100 mL) to obtain 2-(3,4-difluorophenyl)-5-fluoro-4-(methoxycarbonyl)pyridine 1-oxide (311-7A). LC-MS: [M+H] + =284.2.

[0399] A mixture of methylbenzyl((2S,3R)-2-((benzyloxy)methyl)piperidine-3-yl)carbamate (311-7) (17 g, 0.06 mol, 4.0 equivalents) and 2-(3,4-difluorophenyl)-5-fluoro-4-(methoxycarbonyl)pyridine 1-oxide (311-7A) (6 g, approximately 15 mmol, 1.0 equivalent) in DIPEA (40 mL) and DMSO (40 mL) was stirred at 120 °C for 5 hours. The reaction mixture was diluted with H2O (100 mL), extracted with EA (50 mL x 3), washed with saline solution (20 mL), dried over Na2SO4, and concentrated. The residue was purified by combi-flash (EA%=10%~100%) to obtain 5-(3-(benzyl(methoxycarbonyl)amino)-2-((benzyloxy)methyl)piperidine-1-yl)-2-(3,4-difluorophenyl)-4-(methoxycarbonyl)pyridine 1-oxide(311-8). LCMS:[M+H] + =632.5.

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

[0401] A mixture of methyl 5-(3-(benzyl(methoxycarbonyl)amino)-2-((benzyloxy)methyl)piperidine-1-yl)-2-(3,4-difluorophenyl)isonicotinate (311-9) (1500 mg, 2.44 mmol, 1.0 equivalent) and Pd(OH)2 (1500 mg, 100 wt%) in EA (45 mL) was stirred at 60°C and 50 Psi for 18 hours. The reaction mixture was filtered and concentrated. The residue was purified by combi-flash (EA%=0%~50%) to obtain methyl 5-(3-(benzyl(methoxycarbonyl)amino)-2-(hydroxymethyl)piperidine-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)piperidine-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), 2,2-difluoro-2-(fluorosulfonyl)acetic acid (0.113 mL, 1.142 mmol) was added dropwise at 55°C, and the reaction mixture was stirred at 55°C for 2 hours. The reaction mixture was diluted with EA and washed with water and saline. The organic layer was dried over Na2SO4 and concentrated under vacuum to obtain the crude product. The crude product was purified by flash chromatography (elution gradient: 0%~40% ELISA in PE, 30 minutes) to obtain methyl 5-(3-(benzyl(methoxycarbonyl)amino)-2-((difluoromethoxy)methyl)piperidine-1-yl)-2-(3,4-difluorophenyl)isonicotinate (intermediate 311-11). LCMS:[M+H] + =576.3. [Examples]

[0403] Example 1: (R)-9-((5-(3-aminopiperidine-1-yl)-2-(3-fluoro-4-methoxyphenyl)pyridine-4-yl)methyl)-9H-purine-6-amine [ka] A suspension of 5-fluoro-2-(3-fluoro-4-methoxyphenyl)isonicotinaldehyde (intermediate 1-1) (800 mg, 3.21 mmol), (R)-tert-butylpiperidine-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 hours. The mixture was diluted with H₂O (30 mL) and extracted with ELISA (20 mL x 3). The combined organic layers were washed with H₂O (20 mL x 2) and saline solution (20 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography (0-30% ethyl acetate in PE, 30 minutes) to obtain tert-butyl(R)-(1-(6-(3-fluoro-4-methoxyphenyl)-4-formylpyridine-3-yl)piperidine-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-formpyridine-3-yl)piperidine-3-yl)carbamate (1-2) (260 mg, 0.605 mmol) in MeOH (15 mL), sodium borohydride (22.9 mg, 0.605 mmol) was added. The mixture was stirred at room temperature for 30 minutes, 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% ethyl acetate in PE, 30 minutes) to obtain tert-butyl(R)-(1-(6-(3-fluoro-4-methoxyphenyl)-4-(hydroxymethyl)pyridine-3-yl)piperidine-3-yl)carbamate (1-3). 1 1H NMR (400MHz, CD3OD) 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] To a solution of tert-butyl(R)-(1-(6-(3-fluoro-4-methoxyphenyl)-4-(hydroxymethyl)pyridine-3-yl)piperidine-3-yl)carbamate (1-3) (370 mg, 0.857 mmol), intermediate B (300 mg, 0.895 mmol), and PPh3 (704 mg, 2.68 mmol) in THF (15 mL), DEAD (0.425 mL, 2.68 mmol) was added dropwise at 0°C, and the mixture was stirred at room temperature for 1 hour. The mixture was diluted with H2O (20 mL) and extracted with ELISA (20 mL x 3). The combined organic layer was washed with saline solution (20 mL), dried over Na2SO4, filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography (10-50% ELISA in hexane, 40 minutes) to obtain tert-butyl(R)-(tert-butoxycarbonyl)(9-((5-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-2-(3-fluoro-4-methoxyphenyl)pyridine-4-yl)methyl)-9H-purine-6-yl)carbamate(1-4). 1 H NMR(400MHz,CD3OD)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)piperidine-1-yl)-2-(3-fluoro-4-methoxyphenyl)pyridine-4-yl)methyl)-9H-purine-6-yl)carbamate (1-4) (360 mg, 0.481 mmol) in DCM (9 mL), TFA (3 mL) was added. The mixture was stirred at room temperature for 1 hour. The mixture was concentrated under vacuum. The residue was purified by preparative HPLC (basic conditions, NH3H2O%=0.05%, MECN / H2O=0~95%, 12 minutes) to obtain (R)-9-((5-(3-aminopiperidine-1-yl)-2-(3-fluoro-4-methoxyphenyl)pyridine-4-yl)methyl)-9H-purine-6-amine (Example 1). 1 H NMR(400MHz,CD3OD)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] Examples 2 to 21 can be prepared by following the same procedure as described in Example 1. [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)piperidine-1-yl)-2-(3,4-difluorophenyl)pyridine-4-yl)methyl)-9H-purine-6-amine [ka] A mixture of tert-butyl(3-(cyclopropoxymethyl)piperidine-3-yl)carbamate (intermediate 22-7) (300 mg, 1.11 mmol, 1.0 equivalent), methyl 2-(3,4-difluorophenyl)-5-fluoroisonicotinate (intermediate A) (296 mg, 1.11 mmol, 1.0 equivalent), and DIPEA (1.00 g, 7.77 mmol, 7.0 equivalent) in DMSO (3 mL) was stirred at 120 °C for 3 hours under N2. The mixture was quenched with water (200 mL) at 10 °C and extracted with SiO (100 mL x 3 times). The combined organic layer was dried over anhydrous Na2SO4 and concentrated under vacuum. The residue was purified with Combi Flash (20% ELISA in PE) to obtain methyl 5-(3-((tert-butoxycarbonyl)amino)-3-(cyclopropoxymethyl)piperidine-1-yl)-2-(3,4-difluorophenyl)isonicotinate (22-8). 1¹H NMR (400 MHz, CDCl3) 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)piperidine-1-yl)-2-(3,4-difluorophenyl)isonicotinate (22-8) (330 mg, 0.638 mmol, 1.0 equivalent) in THF / MeOH (10 mL, v / v=1 / 1), LiCl (270 mg, 6.38 mmol, 10.0 equivalent), followed by NaBH4 (480 mg, 12.75 mmol, 20.0 equivalent), was added at 15-20°C. The resulting mixture was stirred at 30°C for 2 hours. The reaction product was quenched with water (100 mL) and extracted with ELISA (100 mL x 3). The combined organic layers were dried over anhydrous Na2SO4 and concentrated in vacuum to obtain crude tert-butyl(3-(cyclopropoxymethyl)-1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridine-3-yl)piperidine-3-yl)carbamate (22-9). 1¹H NMR (400 MHz, CDCl3) 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] To a solution of tert-butyl(3-(cyclopropoxymethyl)-1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridine-3-yl)piperidine-3-yl)carbamate (22-9) (300 mg, 0.613 mmol, 1.0 equivalent), intermediate B (310 mg, 0.919 mmol, 1.5 equivalents), and Bu3P (372 mg, 1.84 mmol, 3.0 equivalents) in THF (10 mL), DIAD (372 mg, 1.84 mmol, 3.0 equivalents) was added at 0°C. The mixture was stirred at 20-30°C for 2 hours. The reaction product was concentrated under vacuum. The residue was purified with Combi Flash (30-40% ELISA in PE) to obtain tert-butyl(tert-butoxycarbonyl)(9-((5-(3-(tert-butoxycarbonyl)amino)-3-(cyclopropoxymethyl)piperidine-1-yl)-2-(3,4-difluorophenyl)pyridine-4-yl)methyl)-9H-purine-6-yl)carbamate (22-10). 1H NMR(400MHz,CDCl3)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)piperidine-1-yl)-2-(3,4-difluorophenyl)pyridine-4-yl)methyl)-9H-purine-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 hours. The pH of the reaction mixture was then adjusted to 8-9 with NH3·H2O and extracted with DCM (100 mL x 3). The combined organic layer was dried over anhydrous Na2SO4 and concentrated under vacuum. The residue was purified by preparative HPLC (with 0.1% NH3·H2O as an additive) and SFC to obtain (R)-9-((5-(3-amino-3-(cyclopropoxymethyl)piperidine-1-yl)-2-(3,4-difluorophenyl)pyridine-4-yl)methyl)-9H-purine-6-amine (Example 22). 1H NMR(400MHz,CD3OD)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 to 46 can be prepared by following the same procedure as 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)piperidine-1-yl)-2-(3,4-difluorophenyl)pyridine-4-yl)methyl)-9H-purine-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)piperidine-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 stirred at 120 °C for 8 hours under an N2 atmosphere. The reaction mixture was diluted with water, extracted with ELISA (20 mL x 3), washed with water (20 mL) and saline solution (20 mL), dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by flash chromatography (elution gradient: 0%~30% ethyl acetate in PE, 30 minutes) to obtain methyl 5-(3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoroethyl)piperidine-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)piperidine-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 stirred at room temperature under an N2 atmosphere for 5 hours. The reaction mixture was diluted with water (20 mL), extracted with ELISA (20 mL x 3), washed with water (20 mL) and saline solution (20 mL), dried over anhydrous sodium sulfate, and concentrated to obtain the residue. The residue was purified by flash chromatography (elution gradient: 10%-50% phenylethylamine in hexane, 30 minutes) to obtain tert-butyl(3-(2,2-difluoroethyl)-1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridine-3-yl)piperidine-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)pyridine-3-yl)piperidine-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), DEAD (0.567 mL, 3.58 mmol) was added dropwise at 0°C, and the reaction mixture was stirred at 0°C for 0.5 hours under an N2 atmosphere. The reaction mixture was diluted with water (20 mL), extracted with RINKAN (20 mL x 3), washed with water (10 mL) and saline (20 mL), dried over sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by flash chromatography (elution gradient: 0%~10% MeOH in DCM, 30 minutes) to obtain tert-butyl(tert-butoxycarbonyl)(9-(5-(3-(tert-butoxycarbonyl)amino)-3-(2,2-difluoroethyl)piperidine-1-yl)-2-(3,4-difluorophenyl)pyridine-4-yl)methyl)-9H-purine-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)piperidine-1-yl)-2-(3,4-difluorophenyl)pyridine-4-yl)methyl)-9H-purine-6-yl)carbamate (47-9) (700 mg, 0.875 mmol) in DCM (18 mL), TFA (6 mL) was added, and the reaction mixture was stirred at room temperature under an N2 atmosphere for 2 hours. The reaction mixture was concentrated under vacuum to obtain the crude product. The crude product was purified by preparative HPLC (basic conditions, NH3H2O%=0.05%, MECN / H2O=0~95%, 12 minutes) to obtain a pure racemic product. The racemic product was isolated by SFC to obtain (S)-9-((5-(3-amino-3-(2,2-difluoroethyl)piperidine-1-yl)-2-(3,4-difluorophenyl)pyridine-4-yl)methyl)-9H-purine-6-amine (Example 47). 1 H NMR(400MHz,CD3OD)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 to 77 can be prepared from the corresponding intermediates by following a procedure similar to that 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-purine-9-yl)methyl)-6-(3-fluorophenyl)pyridine-3-yl)-N-methylpiperidine-3-carboxamide [ka]

[0420] To a solution of methyl tert-butyl 3-(methylcarbamoyl)piperidine-3-yl carbamate (intermediate 78-5) (200 mg, 0.777 mmol) and 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 overnight at 120 °C. The reaction mixture was diluted with water, extracted with ethyl acetate, washed with water (20 mL) and saline solution (20 mL), dried over anhydrous sodium sulfate, and concentrated to obtain the residue. The residue was purified by flash chromatography (elution gradient: 0%~30% ethyl acetate in PE, 30 minutes) to obtain methyl 5-(3-(tert-butoxycarbonylamino)-3-(methylcarbamoyl)piperidine-1-yl)-2-(3-fluorophenyl) isonicotinate (78-6). LC-MS:[M+H] + =487.3.

[0421] A mixture of methyl 5-(3-(tert-butoxycarbonylamino)-3-(methylcarbamoyl)piperidine-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 gradually mixed with NaBH4 (232 mg, 6.16 mmol). After the addition, the mixture was stirred at room temperature for 3 hours. Then, water (10 mL) was added to quench the reaction product, it was extracted with ethyl acetate, the combined organic phase was washed with water (20 mL) and saline solution, dried over anhydrous sodium sulfate, and concentrated to obtain the residue. The residue was purified by flash chromatography (elution gradient: 10%-50% ethyl acetate in PE, 30 minutes) to obtain tert-butyl 1-(6-(3-fluorophenyl)-4-(hydroxymethyl)pyridine-3-yl)-3-(methylcarbamoyl)piperidine-3-ylcarbamate (78-7). LC-MS: [M+H] + =459.2.

[0422] To a solution of tert-butyl 1-(6-(3-fluorophenyl)-4-(hydroxymethyl)pyridine-3-yl)-3-(methylcarbamoyl)piperidine-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), DIAD (171 mg, 0.845 mmol) was added dropwise at room temperature under an N2 atmosphere. After addition, the mixture was stirred at room temperature under N2 for 1 hour, quenched with water, and extracted with phenylethylamine. The combined organic phase was work-up under aqueous conditions and concentrated to obtain the residue. The residue was purified by flash chromatography (elution gradient: 10%-70% ethyl acetate in PE, 30 minutes) to obtain tert-butyl(tert-butoxycarbonyl)(9-(5-(3-(tert-butoxycarbonyl)amino)-3-(methylcarbamoyl)piperidine-1-yl)-2-(3-fluorophenyl)pyridine-4-yl)methyl)-9H-purine-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)piperidine-1-yl)-2-(3-fluorophenyl)pyridine-4-yl)methyl)-9H-purine-6-yl)carbamate (78-8) (100 mg, 0.232 mmol) in DCM (9 mL), TFA (3 mL) was added at room temperature. The reaction mixture was stirred at room temperature for 1 hour. The mixture was concentrated under vacuum to obtain the residue. The residue was purified by preparative HPLC (basic conditions, NH3H2O%=0.05%, MEOH / H2O=0~95%, 12 minutes) to obtain 40 mg of pure racemic product. The racemic product was isolated by SFC to obtain (R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(3-fluorophenyl)pyridine-3-yl)-N-methylpiperidine-3-carboxamide (Example 78). 1 H NMR(400MHz,CD3OD)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] Examples 79 to 88 can be prepared by following the same procedure as described in Example 78. [Table 7-1] [Table 7-2] [Table 7-3]

[0425] Example 89: 1-(3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(3,4-difluorophenyl)pyridine-3-yl)piperidine-3-yl)-2,3-dimethylbutan-1-one [ka] DIPEA (724 mg, 5.6 mmol, 4.0 equivalents) was added to a solution of methyl (3-(2,3-dimethylbutanoyl)piperidine-3-yl)carbamate (intermediate 89-6) (367 mg, 1.4 mmol, 1.0 equivalent) and methyl 2-(3,4-difluorophenyl)-5-fluoroisonicotinate (intermediate A) (383 mg, 1.4 mmol, 1.0 equivalent) in DMSO (10 mL). The mixture was stirred at 120 °C for 5 hours under an N2 atmosphere. The mixture was diluted with H2O (20 mL) and extracted with RINKAN (20 mL x 2). The combined organic layers were washed with saline solution (30 mL), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography (30% ethyl acetate in PE) to obtain methyl 2-(3,4-difluorophenyl)-5-(3-(2,3-dimethylbutanoyl)-3-((methoxycarbonyl)amino)piperidine-1-yl) isonicotinate (89-7). 1H NMR(400MHz,CD3OD)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] A mixture of methyl 2-(3,4-difluorophenyl)-5-(3-(2,3-dimethylbutanoyl)-3-((methoxycarbonyl)amino)piperidine-1-yl)isonicotinate (89-7) (416 mg, 0.82 mmol, 1.0 equivalent) in CH3OH (10 mL) was mixed with NaBH4 (312 mg, 8.2 mmol, 10 equivalents) at room temperature. The mixture was stirred at room temperature for 20 minutes. Another NaBH4 (312 mg, 8.2 mmol, 10 equivalents) was added, and the mixture was stirred at room temperature for another 20 minutes. The mixture was diluted with H2O (20 mL) and extracted with ELISA (20 mL x 2). The combined organic layers were washed with saline solution (20 mL), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography (30% ELISA in hexane) to obtain methyl (1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridine-3-yl)-3-(1-hydroxy-2,3-dimethylbutyl)piperidine-3-yl)carbamate (89-8). 1H NMR(400MHz,CD3OD)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] To a solution of methyl (1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridine-3-yl)-3-(1-hydroxy-2,3-dimethylbutyl)piperidine-3-yl)carbamate (intermediate 89-8) (273 mg, 0.57 mmol, 1.0 equivalent), intermediate B (192 mg, 0.57 mmol, 1.0 equivalent), and Bu3P (578 mg, 2.86 mmol, 5.0 equivalents) in THF (15 mL), DIEA (578 mg, 2.86 mmol, 5.0 equivalents) was added dropwise at 0°C under N2. The reaction mixture was then stirred at room temperature under N2 for 1 hour. The mixture was diluted with H2O (20 mL) and extracted with RINKAN (20 mL x 2). The combined organic layers were washed with saline (30 mL), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography (50% ethyl acetate in PE) to obtain tert-butyl(tert-butoxycarbonyl)(9-((2-(3,4-difluorophenyl)-5-(3-(1-hydroxy-2,3-dimethylbutyl)-3-((methoxycarbonyl)amino)piperidine-1-yl)pyridine-4-yl)methyl)-9H-purine-6-yl)carbamate(89-9). 1H NMR(400MHz,CD3OD)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)piperidine-1-yl)pyridine-4-yl)methyl)-9H-purine-6-yl)carbamate (89-9) (279 mg, 0.37 mmol, 1.0 equivalent) and DMP (237 mg, 0.56 mmol, 1.5 equivalents) in DCM (10 ml) was stirred at room temperature for 4 hours. The mixture was quenched with saturated NaHCO3 aqueous solution (30 mL) and extracted with DCM (30 mL x 2). The combined organic layers were washed with saturated NaHCO3 aqueous solution (20 mL x 2) and saline solution (30 mL), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum to obtain the residue. The residue was purified by silica gel column chromatography (50% ethyl acetate in PE) to obtain tert-butyl(tert-butoxycarbonyl)(9-((2-(3,4-difluorophenyl)-5-(3-(2,3-dimethylbutanoyl)-3-((methoxycarbonyl)amino)piperidine-1-yl)pyridine-4-yl)methyl)-9H-purine-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)piperidine-1-yl)pyridine-4-yl)methyl)-9H-purine-6-yl)carbamate (89-10) (280 mg, crude, 0.34 mmol, 1 equivalent) and 10 mL of 35% HBr in acetic acid was stirred at room temperature for 20 hours. LC-MS showed that most of the starting material had been consumed. The reaction mixture was concentrated under vacuum, diluted with H2O (30 mL), and washed with ELISA (30 mL x 3) to remove impurities. The aqueous layer was basicized to pH=11 with NH3·H2O and extracted with ELISA (30 mL x 2). The organic layer was dried over Na2SO4, filtered, and concentrated to obtain the crude product, which was purified by preparative HPLC to obtain 1-(3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(3,4-difluorophenyl)pyridine-3-yl)piperidine-3-yl)-2,3-dimethylbutan-1-one (Example 89). 1 H NMR(400MHz,CD3OD)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] Examples 90 to 112 can be prepared by following a procedure similar to that described in Example 89. [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] Examples 113 and 114: (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(3,4-difluorophenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol and (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(3,4-difluorophenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol [ka] To a solution of tert-butyl(3-(2,2-difluoro-1-hydroxyethyl)piperidine-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 stirred at 120 °C for 8 hours under an N2 atmosphere. The reaction mixture was diluted with water (40 mL) and extracted with ELISA (30 mL x 3). The combined organic layers were washed with water (20 mL x 3) and saline solution (20 mL), dried over anhydrous Na2SO4, filtered, and concentrated to obtain the residue. The residue was purified by flash chromatography (elution gradient: 10% to 50% ethyl acetate in PE, 40 minutes) to obtain methyl 5-(3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidine-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)piperidine-1-yl)-2-(3,4-difluorophenyl)isonicotinate (113-6) (1.2 g, 2.275 mmol) in THF (30 mL) and MeOH (30 mL), lithium chloride (1.929 g, 45.5 mmol) and sodium tetrahydroborate (1.721 g, 45.5 mmol) were added at 0°C, and the reaction mixture was stirred at room temperature under an N2 atmosphere for 4 hours. The reaction mixture was diluted with 30 mL of water and extracted with ELISA (20 mL x 3). The combined organic layers were washed with water (20 mL) and saline solution (20 mL), concentrated, and the crude product was obtained. The crude product was purified by flash chromatography (elution gradient: 20% to 70% ethyl acetate in PE, 40 minutes) to obtain tert-butyl(3-(2,2-difluoro-1-hydroxyethyl)-1-(6-(3,4-difluorophenyl)-4-(hydroxymethyl)pyridine-3-yl)piperidine-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)pyridine-3-yl)piperidine-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), DEAD (0.761 mL, 4.80 mmol) was added dropwise at 0°C. After addition, the reaction mixture was stirred at 0°C for 0.5 hours under an N2 atmosphere. The reaction mixture was diluted with 20 mL of water and extracted with ELISA (20 mL x 3). The combined organic layer was washed with water (20 mL) and saline solution (20 mL), concentrated, and the crude product was obtained. The crude product was purified by flash chromatography (elution gradient: 20%-70% ethyl acetate in PE, 50 minutes) to obtain tert-butyl(tert-butoxycarbonyl)(9-(5-(3-(tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidine-1-yl)-2-(3,4-difluorophenyl)pyridine-4-yl)methyl)-9H-purine-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)piperidine-1-yl)-2-(3,4-difluorophenyl)pyridine-4-yl)methyl)-9H-purine-6-yl)carbamate (113-8) (600 mg, 0.735 mmol) in DCM (30 mL), TFA (10 mL) was added, and the reaction mixture was stirred at room temperature under an N2 atmosphere for 2 hours. The reaction mixture was concentrated under vacuum to obtain the crude product. The crude product was purified by preparative HPLC (basic conditions NH3H2O%=0.05%, MEOH / H2O=0~95%, 10 min) to obtain 330 mg of pure racemic product. The racemic product was isolated by SFC to obtain (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(3,4-difluorophenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol (Example 113) and (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(3,4-difluorophenyl)pyridine-3-yl)-2,2-difluoroethane-1-ol (Example 114).

[0435] Example 113: 1 H NMR(CD3OD,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) ,5.91-6.19(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: 1H NMR(CD3OD-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] Examples 115 to 155 can be prepared by following the same procedure as described in Examples 113 and 114. [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] Examples 156 and 157: (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2,4,5-trifluorophenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol and (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2,4,5-trifluorophenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol [ka] Step 1. DIAD (509 mg, 2.516 mmol, 1.5 equivalents) was added under nitrogen at 0°C to a solution of 1-(3-amino-1-(4-(hydroxymethyl)-6-(2,4,5-trifluorophenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol (intermediate 156-2) (700 mg, 1.677 mmol, 1.0 equivalent), intermediate B (80 mg, 0.240 mmol, 1.5 equivalents), and n-Bu3P (509 mg, 2.516 mmol, 1.5 equivalents) in anhydrous THF (7 mL). The reaction mixture was stirred at 0°C for 30 minutes. The reaction mixture was concentrated under vacuum, and the residue was purified by preparative HPLC (bases) to obtain intermediate 156-3. 1H NMR(400MHz,CDCl3)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), TFA (5 mL) was added, and the reaction mixture was stirred at 20°C for 3 hours. The reaction mixture was concentrated under vacuum, basicized to pH=8 with ammonium hydroxide, and concentrated under vacuum. The residue was purified by preparative HPLC (base) and SFC to obtain (R)-1-((R)-3-amino-1-(4-(6-amino-9H-purine-9-yl)methyl)-6-(2,4,5-trifluorophenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol (Example 156) and (S)-1-((R)-3-amino-1-(4-(6-amino-9H-purine-9-yl)methyl)-6-(2,4,5-trifluorophenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol (Example 157).

[0440] Example 156: 11H 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] Examples 158 and 159 were prepared following the same procedure as for the preparations in Examples 156 and 157 (Step 2). [Table 10]

[0443] Examples 160 and 161: (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2,5-difluoro-4-methoxyphenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol and (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2,5-difluoro-4-methoxyphenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol [ka] To a solution of tert-butyl(tert-butoxycarbonyl)(9-((5-(3-((tert-butoxycarbonyl)amino)-3-(2,2-difluoro-1-hydroxyethyl)piperidine-1-yl)-2-(2,5-difluoro-4-methoxyphenyl)pyridine-4-yl)methyl)-9H-purine-6-yl)carbamate (intermediate 160-3) (200 mg, 0.237 mmol) in DCM (18 mL), TFA (36 mL) was added, and the reaction mixture was stirred at room temperature under an N2 atmosphere for 30 minutes. The reaction mixture was concentrated under vacuum to obtain the crude product. The crude product was purified by preparative HPLC and SFC to obtain (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2,5-difluoro-4-methoxyphenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol (Example 160) and (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2,5-difluoro-4-methoxyphenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol (Example 161).

[0444] Example 160: 1H NMR(400MHz,CD3OD)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,CD3OD)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] Examples 162 and 163: (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2-(difluoromethyl)-4-methoxyphenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol and (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2-(difluoromethyl)-4-methoxyphenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol [ka] DEAD (0.061 mL, 0.386 mmol) was added to a solution of intermediate 162-2 (57 mg, 0.129 mmol), intermediate B (43.1 mg, 0.129 mmol), and PPh3 (101 mg, 0.386 mmol) in THF (5 mL). The reaction mixture was stirred at 0°C for 30 minutes under an N2 atmosphere. The reaction mixture was quenched with water, extracted with RINKAN, washed with water and saline, dried over sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by flash chromatography (elution gradient: 0%~10% MeOH in DCM, 30 minutes) to obtain the intermediate. LC-MS: [M+H] + =761.

[0447] To a solution of the intermediate (200 mg, 0.263 mmol) in DCM (18 mL), TFA (36 mL, 467 mmol) was added, and the reaction mixture was stirred at room temperature under an N2 atmosphere for 30 minutes. The crude product was purified by preparative HPLC (basic conditions, NH3H2O%=0.05%, MeOH / H2O=0~95%, 10 min) to obtain a pure product, which was then further purified to obtain (R)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2-(difluoromethyl)-4-methoxyphenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol (Example 162) and (S)-1-((R)-3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(2-(difluoromethyl)-4-methoxyphenyl)pyridine-3-yl)piperidine-3-yl)-2,2-difluoroethane-1-ol (Example 163).

[0448] Example 162: 1H NMR(400MHz,CD3OD)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,CD3OD)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] Examples 164-167 were prepared from intermediate B and the corresponding intermediates following the same procedure as in the preparations of Examples 162 and 163. [Table 11-1] [Table 11-2] [Table 11-3]

[0451] Examples 168 to 176 were prepared from the corresponding intermediates following the same procedure as in the preparations of Examples 168 and 169. [Table 12-1] [Table 12-2] [Table 12-3] [Table 12-4]

[0452] Example 177: 9-((5-(3-amino-3-(6-methylpyridine-2-yl)piperidine-1-yl)-2-(3,4-difluorophenyl)pyridine-4-yl)methyl)-9H-purine-6-amine [ka] A solution of tert-butyl(tert-butoxycarbonyl)(9-((2-(3,4-difluorophenyl)-5-(3-((methoxycarbonyl)amino)-3-(6-methylpyridine-2-yl)piperidine-1-yl)pyridine-4-yl)methyl)-9H-purine-6-yl)carbamate (177-7) (700 mg, 0.89 mmol, 1.0 equivalent) in HBr / AcOH (33%, 10 mL) was stirred at 25-30°C for 16 hours. The reaction mixture was concentrated under vacuum. The residue was purified by preparative HPLC (with 0.1% NH4HCO3 as an additive) to obtain 9-((5-(3-amino-3-(6-methylpyridine-2-yl)piperidine-1-yl)-2-(3,4-difluorophenyl)pyridine-4-yl)methyl)-9H-purine-6-amine (Example 177). 1H NMR(400MHz,DMSO-d6)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] Examples 178 to 284 were prepared following the same procedure as in Example 177 and the preparation of the corresponding intermediates. [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 [Table 13-27] [Table 13-28] [Table 13-29] [Table 13-30] [Table 13-31] [Table 13-32] [Table 13-33] [Table 13-34] [Table 13-35] [Table 13-36] [Table 13-37] [Table 13-38] [Table 13-39]

[0454] Examples 285 to 291 were prepared from the corresponding intermediates following a procedure similar to that 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-purine-9-yl)methyl)-6-(3,4-difluorophenyl)pyridine-3-yl)-3-(2,2-difluoroethyl)piperidine-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), K2CO3 (215 mg, 1.556 mmol) was added at room temperature under an N2 atmosphere, and the reaction mixture was stirred at room temperature for 18 hours. The reaction mixture was diluted with water and extracted with RINKAN (20 mL x 3). The combined organic phase was washed with water (20 mL x 3) and saline solution (20 mL), dried over anhydrous sodium sulfate, and concentrated under vacuum to obtain the residue. The residue was purified by flash chromatography (elution gradient: 0%~10% MeOH in DCM, 30 minutes) to obtain 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 at room temperature under an N2 atmosphere, and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under vacuum to obtain the residue. The residue was purified by preparative HPLC (basic conditions, NH3H2O%=0.05%, MeCN / H2O=0~95%, 12 min) to obtain 3-amino-1-(4-((6-amino-9H-purine-9-yl)methyl)-6-(3,4-difluorophenyl)pyridine-3-yl)-3-(2,2-difluoroethyl)piperidine-4-ol (Example 301). 1 H NMR(400MHz,CD3OD)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] Examples 302 to 310 were prepared following the same procedure as in Example 47 and the preparation of the corresponding intermediates. [Table 15-1] [Table 15-2]

[0458] Example 306: 9-((5-((3R,5R)-3-amino-5-(fluoromethyl)piperidine-1-yl)-2-(3-fluoro-4-methoxyphenyl)pyridine-4-yl)methyl)-9H-purine-6-amine [ka] A mixture of tert-butyl(tert-butoxycarbonyl)(9-((5-((3R,5R)-3-((tert-butoxycarbonyl)amino)-5-(fluoromethyl)piperidine-1-yl)-2-(3-fluoro-4-15methoxyphenyl)pyridine-4-yl)methyl)-9H-purine-6-yl)carbamate (intermediate 306-9) (0.15 g, 0.19 mmol) in DCM (1 mL) was mixed with HCl / dioxane (10.0 mL, 4 M), and the resulting mixture was stirred at 25°C for 4 hours. The mixture was concentrated under vacuum. The residue was purified by preparative HPLC (with 0.1% NH4HCO3 as an additive) to obtain 9-((5-((3R,5R)-3-amino-5-(fluoromethyl)piperidine-1-yl)-2-(3-fluoro-4-methoxyphenyl)pyridine-4-yl)methyl)-9H-purine-6-amine (Example 306). 1 H NMR(400MHz,DMSO-d6):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] Examples 307 to 310 were prepared following the same procedure as for the preparation of Example 306 and the corresponding intermediate. [Table 16]

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

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

[0462] Example 322 was prepared following the same procedure as in Example 89 and the preparation of the corresponding intermediate. 1 H NMR(400MHz,CD3OD)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] Examples 323 to 325 were prepared following the same procedure as for the preparation of Example 1 and the corresponding intermediate. [Table 18]

[0464] Biological assays The compounds of the present invention can be evaluated for their ability to inhibit NSD2 using the assays described below and other assays 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). ...

Claims

1. Compound of formula (I) 【Chemistry 1】 or its enantiomer, enantiomer mixture, or pharmaceutically acceptable salt, wherein in the formula, A is N or CR ９ And R ９ is hydrogen or halo, L is a combination or C １－４ It is alkylene, R １ is H, or R １ and R ２ However, together with NH, it forms a 5-8 membered heterocycline containing 1-2 heteroatoms selected from N, O, and S as ring members, and the 5-8 membered heterocycline is either unsubstituted or substituted with an oxo substituent. R ２ teeth, (i) hydrogen, -C １－６ alkyl, -halo C １－６ alkyl, -hydroxy C １－６ alkylene, -hydroxyhalo C １－６ alkylene, -C １－６ alkoxy C １－６ alkylene, -halo C １－６ alkoxy C １－６ alkylene, or -C ３－８ cycloalkoxy (C １－６ alkyl), (ii) Cyano, -Cyano C １－６ Alkylene, -C １－６ Alkylthio C １－６ Alkyl, -C ２－６ Alkenil, -Halo C ２－６ Alkenyl, -C ２－６ Alkinyl, -C １－４ Alkyl SOC １－４ Alkyl, -C １－４ Alkyl SO 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 C ３－７ The cycloalkyl, 5-6 membered heterocyclyl, or 5-6 membered heteroaryl 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 It is alkyl, R １３ is hydrogen, -C １－６ It is either an alkyl group or a 5-6 membered heterocycle, or R １２ and R １３ These combine to form a 5-6 membered heterocycle, and the 5-6 membered heterocycle is either unsubstituted or １－４ (substituted with alkyl) (v) 5-6 member heterocyclyl C ０－６ Alkyl or 5-6 member heterocycline (halo C) １－４ Alkyl) (wherein each of the heterocyclyl groups is either unsubstituted or substituted with an oxo), (vi) 5-9 member heteroaryl C ０－６ Alkyl or 5-9 member heteroaryl (halo C) １－４ Alkyl) (wherein each of the heteroaryl groups is unsubstituted or -C １－４ Alkyl, -halo C １－４ Alkyl, -hydroxyC １－４ Alkylene, -C １－４ Alkoxy, -halo C 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 Selected from the group consisting of (substituted by cycloalkyl), R ３ａ , R ３ｂ , R ４ａ , R ４ｂ , R ５ａ , R ５ｂ , R ６ａ , and R ６ｂ These are independently hydrogen, halo, cyano, and hydroxyl. -C １－６ alkyl, halo-C １－６ alkyl, hydroxy-C １－６ alkylene, C １－６ alkoxy, C １－６ alkoxy C １－６ alkylene, halo-C １－６ alkoxy C １－６ alkylene, hydroxyhalo-C 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 form an oxo substituent, R ７ H, -C １－４ Alkoxy, Halo, C １－４ Alkyl, unsubstituted, or halo-substituted 3- to 8-membered heterocyclyl, R ８ 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- to 9-membered heteroaryl C containing 1-3 heteroatoms selected from N, O, and S. ０－６ It is alkyl, R ８ is either unsubstituted or has 1 to 3 R １７ Replaced by, R １７ is halo, hydroxy, cyano, -C １－６ Alkyl, -halo C １－６ Alkyl, -C １－６ Alkoxy, -halo C 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 These are, independently, hydrogen or -C １－４ It is alkyl, R １８ is, -C １－４ Alkyl, or -C １－４ It is a haloalkyl, A compound of formula (I), or an enantiomer thereof, an enantiomer mixture thereof, or a pharmaceutically acceptable salt thereof, wherein p, q, r, s, and t are independently between 0 and 4.

2. A is N, the compound of formula (I) according to claim 1, or an enantiomer thereof, an enantiomer mixture thereof, or a pharmaceutically acceptable salt thereof.

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

4. R ６ａ However, it is hydrogen or halo, R ６ｂ However, hydrogen, -halo C １－６ Alkoxy C １－６ Alkylene, -hydroxyC １－６ Alkylene, -Hydroxyhalo C 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 However, independently, hydrogen or -C １－４ It 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, enantiomer mixture thereof, or a pharmaceutically acceptable salt thereof, wherein together they form an oxo substituent.

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

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, wherein the compound is hydrogen, or a pharmaceutically acceptable salt thereof.

7. The compound of formula (I) according to claim 1, wherein the compound is the compound of formula (II), 【Chemistry 2】 or its enantiomer, enantiomer mixture, or pharmaceutically acceptable salt.

8. R ２ However, hydrogen, -C １－６ Alkyl, - HaroC １－６ Alkyl, -hydroxyC １－６ Alkylene, -Hydroxyhalo C １－６ Alkylene, -C １－６ Alkoxy C １－６ Alkilen, - HaroC １－６ Alkoxy C １－６ Alkylene, or -C ３－８ Cycloalkoxy (C １－６ A compound according to any one of claims 1 to 7, which is alkyl, or an enantiomer thereof, a mixture of enantiomers, or a pharmaceutically acceptable salt thereof.

9. R ２ The compound according to any one of claims 1 to 7, wherein the compound is hydrogen, 2,2-difluoroethyl, 2-methyl-propane-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 thereof, an enantiomer mixture thereof, or a pharmaceutically acceptable salt thereof.

10. R ２ but, 【Transformation 3】 The compound according to any one of claims 1 to 7, or its enantiomer, enantiomer mixture, or pharmaceutically acceptable salt.

11. R ２ However, -C １－４ Alkylcarbonyl, - (CR ａ R ｂ ) ｐ -C(O)-OR １０ (In the formula, R １０ However, hydrogen or -C １－４ (It is alkyl.) -C(O)-(CR ａ R ｂ ) ｑ -R １１ (In the formula, R １１ However, C ３－６ They are cycloalkyl, 5-6 membered heterocyclyl, or 5-6 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 (in the formula, R １２ However, hydrogen or -C １－６ It is alkyl, R １３ However, hydrogen, -C １－６ Alkyl, or a 5-6 membered heterocycle, R １２ and R １３が Together they form a 5-6 membered heterocycle, and the 5-6 membered heterocycle is either unsubstituted or -C １－４ (It is substituted with alkyl) R ａ , R ｂ , and R １１ However, independently, hydrogen or -C １－４ It is alkyl, A compound according to any one of claims 1 to 7, or an enantiomer thereof, an enantiomer mixture thereof, or a pharmaceutically acceptable salt thereof, wherein p, q, and r are independently 0 to 2.

12. R ２ but, Pyridine-2-yl, (pyridine-2-yl)methyl, or (pyridine-2-yl)ethyl (wherein the formula, the pyridine-2-yl is independently unsubstituted or has 1-2 -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 Alkilen, -Halo C １－４ Alkoxy, halo, or -C ３－７ (Substituted with cycloalkyl groups) Thiazole-4-yl, thiazole-2-ylmethyl, oxazole-2-yl, oxazole-2-ylmethyl, pyridazine-3-yl, pyridazine-3-ylmethyl, pyrazine-2-yl, pyrazine-2-ylmethyl, pyrimidine-4-yl, pyrimidine-4-ylmethyl (each of these independently is either unsubstituted or -C １－４ (Substituted with alkyl or halo) 1H-pyrazole-3-yl, (1H-pyrazole-3-yl)methyl, 1-methyl-1H-pyrazole-4-yl, (1,2,3-triazole-4-yl), isoxazole-3-yl (each of these independently unsubstituted or -C) １－４ Alkyl, halo, or -C ３－７ (Substituted with cycloalkyl groups) 1,4-dioxan-2-yl, Pyridine-2(1H)-onyl, and A compound selected from pyridine-1-oxide-2-yl, according to any one of claims 1 to 7, or an enantiomer thereof, an enantiomer mixture thereof, or a pharmaceutically acceptable salt thereof.

13. R ８ The compound according to any one of claims 1 to 12, wherein the compound is cyclopropyl, cyclobutyl, cyclohexyl, or azetidinil, or an enantiomer, enantiomer mixture thereof, or a pharmaceutically acceptable salt thereof.

14. R ８ The compound according to any one of claims 1 to 12, wherein the compound is 2,3-dihydrobenzofuran-5-yl, 1,3-dihydroisobenzofuran-5-yl, benzo[d][1,3]dioxol-5-yl, imidazo[1,2-a]pyridine-6-yl, or pyrazolo[1,5-a]pyridine-6-yl, or an enantiomer thereof, an enantiomer mixture, or a pharmaceutically acceptable salt thereof.

15. R ８ However, it is either unsubstituted or has one or two halos, -C １－６ Alkyl, or -C １－６ Pyridyls substituted with alkoxy; unsubstituted, or with one or two halos or -C １－６ Alkyl-substituted, 1H-indazole-5-yl; or unsubstituted, or -C １－６ A compound according to any one of claims 1 to 12, which is alkyl-substituted 1H-benzo[d]-imidazole-6-yl, or an enantiomer thereof, an enantiomer mixture, or a pharmaceutically acceptable salt thereof.

16. R ８ However, 1 to 3 R １７ It is a phenyl substituted with, R １７ However, halo, hydroxy, cyano, -C １－６ Alkyl, -halo C １－６ Alkyl, -C １－６ Alkoxy, -halo C 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 However, independently, hydrogen or -C １－４ It is alkyl, R １８ However, C １－４ It is a haloalkyl, A compound according to any one of claims 1 to 12, or an enantiomer thereof, an enantiomer mixture thereof, or a pharmaceutically acceptable salt thereof, wherein t is 0 to 1.

17. R ８ However, 1 to 3 R １７ A phenyl substituted with R １７ However, halo, hydroxy, -C １－６ Alkyl, -halo C １－６ Alkyl, -C １－６ Alkoxy, or -halo C １－６ A compound according to any one of claims 1 to 12, which is an alkoxy, or an enantiomer thereof, an enantiomer mixture thereof, or a pharmaceutically acceptable salt thereof.

18. The compound according to claim 1, or an enantiomer, enantiomer mixture, or pharmaceutically acceptable salt thereof, wherein the compound is selected from the compounds of Table 2 or pharmaceutically acceptable salts thereof.

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

20. A combination comprising a compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof, and one or more therapeutic agents.

21. The combination according to claim 20, wherein one or more therapeutic activators are anticancer agents, analgesics, or anti-inflammatory agents.

22. A method for 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 pharmaceutically acceptable salt thereof.

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

24. The method according to 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 lymphoma, or pulmonary hypertension.

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

26. Use of the compound according to any one of claims 1 to 18, optionally combined 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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