Small molecule modulators of alveolar type 2 cell proliferation for the treatment of pulmonary diseases

Abstract

The present disclosure relates to compounds that inhibit dipeptidyl peptidase IV (DPP4) and pharmaceutical compositions thereof.The compounds selectively promote the proliferation of alveolar type 2 cells (AEC2) and are useful in therapeutic methods for treating lung diseases whose etiology is derived from, for example, epithelial degeneration and maladaptive remodeling, such as idiopathic pulmonary fibrosis (IPF), acute respiratory distress syndrome (ARDS) and infantile respiratory distress syndrome (IRDS).

Description

[Technical field]

[0001] This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 139,956, filed January 21, 2021, which is incorporated in its entirety as if fully set forth herein. [Background technology]

[0002] Pharmacological stimulation of lower airway repair has great potential for treating a variety of conditions in which alveolar destruction and maladaptive remodeling are the cause of disease. Alveoli, the primary unit of gas exchange in mammals, are composed of two epithelial cell types: the large squamous alveolar type 1 cells (AEC1), which provide the surface area for gas exchange, and the cuboidal alveolar type 2 cells (AEC2), which secrete surfactant. 1 Furthermore, AEC2s have been identified as the primary progenitor cell type responsible for regenerating the alveolar epithelium. 2 AEC2s undergo clonally expanding throughout adulthood and divide asymmetrically to give rise to AEC1s and AEC2s. 2 It has further been demonstrated that idiopathic pulmonary fibrosis (IPF) is caused by an exhaustion of the stem cell potential of AEC2. 3 Decreased AEC2 proliferation leads to denudation of the alveolar basement membrane, which ultimately promotes colonization of the lower airways by hyperplastic upper airway-derived epithelial cells and extracellular matrix-secreting myofibroblasts. 3 Furthermore, it has been demonstrated that restoring AEC2 proliferation by treatment with exogenous factors (IL-6 or hyaluronan) inhibits disease severity in a mouse model of IPF. 4 In addition to IPF, acute respiratory distress syndrome (ARDS), an acute loss of alveolar epithelial barrier function, is caused by injury to and a lack of reparative growth by AEC2 cells. 5 . Summary of the Invention

[0003] The present disclosure provides, in various embodiments, compounds useful for promoting specific proliferation of AEC2 relative to other cell types, for example, in the lung. In some embodiments, the compound is of formula (I) or a pharmaceutically acceptable salt thereof: [ka]

[0004] In formula (I), each --- represents a single bond which, if present, forms a fused cyclopropyl ring.

[0005] L 1A is -NHCH2- or CH(NH2)-.

[0006] X 1 is selected from -O-, -S-, -S(O)-, S(O)2- and -NH-. 1B is C2-C 12 -alkyl, wherein one or more -CH2- are independently replaced by a moiety selected from -O-, -C(O)-, and -NH-. 1 H, C6-C 10 -aryl and 5-10 membered heteroaryl, wherein 1-4 heteroaryl members are independently selected from N, O and S.

[0007] The subscript m1 is Z 1 is H, then 0; Z 1 is an integer that is 1 if is other than H. The subscript n1 is an integer selected from 0, 1, 2, and 3.

[0008] R 1 , H, C1-C 10 -Alkyl and -C1-C 10 -Alkyl-(C6-C 10 -aryl), optionally substituted with 1 to 6 -OH. 2 is C1-C substituted with 1 to 6 -OH. 10 -alkyl.

[0009] Alternatively, according to a further embodiment, the compound is of formula (II) or a pharma- ceutically acceptable salt thereof: [ka]

[0010] In formula (II), W is CH or N.

[0011] The subscript o is an integer selected from 1, 2, and 3.

[0012] R 3 is C1-C6-alkyl, C1-C6-hydroxyalkyl and -(CH2CH2O) x H, where x is an integer selected from 1, 2, 3, 4 and 5. 4 is C2-C8-alkynyl.

[0013] R 5a , R 5b , R 5c and R 5d is H, C1-C6-alkyl, halo, -NR A R B (R A and R B H and C1-C 10 -alkyl), -C(O)OH, -B(OH), -C(O)NR A R B , -C(O)OR A and -C(O)-L 2 -Z 2 -[(CH2) n2 -NR 6 R 7 ] m2 are independently selected from L 2 is C2-C 12 -alkyl, wherein one or more -CH2- are independently replaced by a moiety selected from -O-, -C(O)-, and -NH-. 2 H, C6-C 10-aryl and 5-10 membered heteroaryl, where 1-4 heteroaryl members are independently selected from N, O and S. 5a , R 5b , R 5c and R 5d At least one of is other than H.

[0014] R 6 , H, C1-C 10 -Alkyl and -C1-C 10 -Alkyl-(C6-C 10 -aryl), optionally substituted with 1 to 6 -OH. 7 is C1-C substituted with 1 to 6 -OH. 10 -alkyl.

[0015] The subscript m2 is Z 1 is H, then 0; Z 1 is an integer that is 1 if is other than H. The subscript n2 is an integer selected from 0, 1, 2, and 3.

[0016] Furthermore, when W is CH, R 5a and R 5d is not selected from -C(O)OH, -C(O)OMe, and -C(O)OEt.

[0017] Alternatively, according to a further embodiment, the compound is of formula (III) or a pharma- ceutically acceptable salt thereof: [ka]

[0018] In formula (III), X 3 is -O- or -NH-. 3 is a bond or C2-C 12 -alkyl, wherein one or more -CH2- are independently replaced by a moiety selected from -O-, -C(O)-, and -NH-. 3 H, -N3, C6-C 10-aryl, 5-10 membered heteroaryl, where 1-4 heteroaryl members are independently selected from N, O and S, and (3-14 membered heterocycloalkyl, where 1-4 ring members are independently selected from N, O and S). Heteroaryl and heterocycloalkyl are optionally substituted with 1-6 substituents selected from the group consisting of halo, NO, OH, CN and C-C-haloalkyl.

[0019] The subscript m3 is Z 3 is H or -N3, then it is 0, and Z 3 is other than H or -N3. The subscript n3 is an integer selected from 0, 1, 2, and 3.

[0020] R 8 , H, C1-C 10 -Alkyl or -C1-C 10 -Alkyl-(C6-C 10 -aryl), optionally substituted with 1 to 6 -OH. 9 is C1-C substituted with 1 to 6 -OH. 10 -alkyl. R 10 is C1-C6-haloalkyl. 11 is independently selected from H, C1-C6-alkyl and halo.

[0021] The subscript o3 is an integer selected from 0, 1, 2, and 3. The subscript p3 is an integer selected from 0, 1, 2, and 3. The subscript q3 is an integer selected from 0, 1, 2, and 3.

[0022] Compounds of the present disclosure do not include any of the following compounds: [Table 1] TIFF2024504328000005.tif212144TIFF2024504328000006.tif50148

[0023] In another embodiment, the present disclosure provides a pharmaceutical composition comprising a compound described herein or a pharma- ceutically acceptable salt thereof.

[0024] The present disclosure also provides, in one embodiment, a method for selectively increasing proliferation of cuboidal alveolar type 2 (AEC2) cells in a subject in need thereof, or for restoring decreased proliferation of AEC2 cells in a subject in need thereof, comprising administering to the subject a compound as described herein, or a pharma- ceutically acceptable salt thereof.

[0025] In one embodiment, the present disclosure provides a method of inhibiting dipeptidyl peptidase IV (DPP4) in a subject in need thereof, comprising administering to the subject a compound described herein or a pharma- ceutically acceptable salt thereof.

[0026] Another embodiment of the present disclosure is a method for treating a pulmonary disease in a subject suffering therefrom, comprising administering to the subject a compound as described herein or a pharma- ceutically acceptable salt thereof.

[0027] Also provided in various embodiments are compounds as described herein, or pharma- ceutically acceptable salts thereof, for use in selectively increasing proliferation of cuboidal alveolar type 2 (AEC2) cells in a subject in need thereof, or for reversing decreased proliferation of AEC2 cells in a subject in need thereof.

[0028] In a further embodiment, the present disclosure provides a compound described herein, or a pharma- ceutically acceptable salt thereof, for use in inhibiting dipeptidyl peptidase IV (DPP4) in a subject in need thereof.

[0029] In yet an additional embodiment, the disclosure provides a compound described herein, or a pharma- ceutically acceptable salt thereof, for use in treating a pulmonary disease in an affected subject. [Brief description of the drawings]

[0030] [Figure 1] AEC2 proliferation-concentration curves for compound 46. [Diagram 2] Comparison of the pharmacokinetics of lilagliptin and compound 46 administered IT at 2 mg / kg in mice [Figure 3A-3E] Compound 46 demonstrated efficacy in a mouse bleomycin-induced pulmonary fibrosis model. (A) Body weight measurements and dosing schedule used for intratracheal administration of Compound 46 (0.5 mg / kg every 4 days). BALF (B), fibrosis area measurements (C), modified Ashcroft score (D), and representative Masson's trichrome stained histological slides at the end of the study (E); **, P<0.005; ***, P<0.0005. [Figure 4A-4D] Compound 46 shows synergistic effects in combination with the standard of care IPF drug nintedanib in a bleomycin-induced fibrosis model in mice. (A) Treatment legend and body weight measurements of bleomycin-induced pulmonary fibrosis model mice. Compound 46, 0.5 mg / kg IT, E4D; BALF protein levels (B), modified Ashcroft score (C), and representative Masson's trichrome stained histological slides at the end of the study (D). *, P<0.05; ***; P<0.005; ***, P<0.0005; NS=not statistically significant. [Figure 5A-5C] Compound 46 selectively increased AEC2 in mice. (A) UMAP plot showing multiple populations of cells identified in mouse lungs, highlighting key populations of interest. (B) UMAP plot showing which cells express a transcription profile consistent with a proliferative state. Other populations besides proliferating AEC2 are immune cells. (C) Quantification of total proliferating cells at the indicated time points after treatment with Compound 46. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0031] The present disclosure satisfies a long-standing need for drug-like compounds that stimulate the repair proliferation of lung stem cell and progenitor cell populations. The compounds of the present disclosure promote the specific proliferation of AEC2s compared to other cell types in the lung (e.g., lung fibroblasts), thereby exhibiting disease-modifying efficacy in several lower airway diseases. In addition, the compounds are useful as inhibitors of dipeptidyl peptidase IV (DPP4).

[0032] definition "Alkyl" refers to straight or branched chain hydrocarbyl containing from 1 to about 20 carbon atoms. For example, an alkyl can have from 1 to 10 carbon atoms or from 1 to 6 carbon atoms. Exemplary alkyls include straight chain alkyl groups such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, and the like, as well as branched chain isomers of straight chain alkyl groups such as, but not limited to, -CH(CH3)2, -CH(CH3)(CH2CH3), -CH(CH2CH3)2, -C(CH3)3, -C(CH2CH3)3, -CH2CH(CH3)2, -CH 2CH(CH3)(CH2CH3), -CH2CH(CH2CH3)2, -CH2C(CH3)3, -CH2C(CH2CH3)3, -CH(CH3)CH(CH3)(CH2CH3), -CH2CH2CH(CH 3)2, -CH2CH2CH(CH3)(CH2CH3), -CH2CH2CH(CH2CH3)2, -CH2CH2C(CH3)3, -CH2CH2C(CH2CH3)3, -CH(CH3)CH2CH(CH3) 2、 and the like. Alkyl groups thus include primary alkyl groups, secondary alkyl groups, and tertiary alkyl groups. Alkyl groups can be unsubstituted or substituted with one or more substituents described herein.

[0033] The term "haloalkyl" refers to an alkyl, as defined herein, that is substituted with one, two, three, four, five, or six halo. An exemplary haloalkyl is -CF3.

[0034] The terms "halogen", "halide" and "halo" each refer to -F or fluoro, -Cl or chloro, -Br or bromo, or -I or iodo.

[0035] "Alkyne" or "alkynyl" refers to a straight or branched chain unsaturated hydrocarbon having the indicated number of carbon atoms and at least one triple bond. Examples of (C2-C8)alkynyl groups include, but are not limited to, acetylene, propyne, 1-butyne, 2-butyne, 1-pentyne, 2-pentyne, 1-hexyne, 2-hexyne, 3-hexyne, 1-heptyne, 2-heptyne, 3-heptyne, 1-octyne, 2-octyne, 3-octyne, and 4-octyne. Alkynyl groups may be unsubstituted or substituted with one or more substituents described herein.

[0036] "Aryl", when used alone or as part of another term, means an optionally fused carbocyclic aromatic group, e.g., C-C, having the specified number of carbon atoms, or, if no number is specified, up to 14 carbon atoms. 14 -Aryl or C6-C 10 -aryl. Exemplary aryl groups are phenyl, naphthyl, biphenyl, phenanthrenyl, naphthacenyl, and the like (see, for example, Table 7-2 in Lang's Handbook of Chemistry (Dean, JA, ed), 13th Edition). An exemplary aryl is phenyl. An "aryl" may be fused with a cycloalkyl ring as defined herein. An aryl group may be unsubstituted or substituted with one or more substituents as described herein.

[0037] The term "heteroatom" refers to N, O, and S. Compounds of the present disclosure containing N or S atoms may be optionally oxidized to the corresponding N-oxide, sulfoxide, or sulfone compounds.

[0038] "Heteroaryl," alone or in combination with any other moiety described herein, refers to a monocyclic aromatic ring structure containing 5-10, e.g., 5 or 6, ring atoms, or a bicyclic aromatic group having 8-10 atoms and containing one or more, e.g., 1-4, 1-3, or 1-2, heteroatoms independently selected from the group consisting of O, S, and N. Heteroaryl is also intended to include oxidized S or N, e.g., sulfinyl, sulfonyl, and N-oxide of a tertiary ring nitrogen. A carbon or heteroatom is the point of attachment of the heteroaryl ring structure, which results in a stable compound. Examples of heteroaryl groups include, but are not limited to, pyridinyl, pyridazinyl, pyrazinyl, quinoxalyl, indolizinyl, benzo[b]thienyl, quinazolinyl, purinyl, indolyl, quinolinyl, pyrimidinyl, pyrrolyl, pyrazolyl, oxazolyl, thiazolyl, thienyl, isoxazolyl, oxathiadiazolyl, isothiazolyl, tetrazolyl, imidazolyl, triazolyl, furanyl, benzofuryl, and indolyl. Heteroaryl groups can be unsubstituted or substituted with one or more substituents described herein.

[0039] "Heterocycloalkyl" refers to a saturated or partially unsaturated non-aromatic monocyclic, bicyclic, tricyclic or polycyclic ring system, optionally spiro-fused, having 3-14, e.g., 3-6, atoms, in which 1-3 carbon atoms in the ring are replaced by O, S or N heteroatoms. Heterocycloalkyls may be fused with 5-6 ring-membered aryl or heteroaryls, and include oxidized S or N, e.g., sulfinyl, sulfonyl and N-oxide of a tertiary ring nitrogen. The attachment point of the heterocycloalkyl ring is at a carbon or heteroatom, so that a stable ring is maintained. Examples of heterocycloalkyl groups include, but are not limited to, morpholino, tetrahydrofuranyl, dihydropyridinyl, piperidinyl, pyrrolidinyl, piperazinyl, dihydrobenzofuryl and dihydroindolyl. Heterocycloalkyl groups may be unsubstituted or substituted with one or more substituents as described herein.

[0040] The terms "nitrile" and "cyano" may be used interchangeably and refer to a -CN group that is attached to carbon atoms of heteroaryl, aryl and heterocycloalkyl rings.

[0041] The term "oxo" refers to an =O atom bonded to an atom that is part of a saturated or unsaturated moiety. Thus, the =O atom can be bonded to a carbon, sulfur, or nitrogen atom that is part of a cyclic or acyclic moiety.

[0042] "Hydroxyl" or "hydroxy" refers to the group --OH.

[0043] The substituent -CO2H represents a bioisosteric substitution, e.g. [ka]

[0044] and the like, where R is R as defined herein. A See, for example, The Practice of Medicinal Chemistry (Academic Press: New York, 1996), p. 203.

[0045] The compounds described herein may exist in various isomeric forms, including, for example, configurational, geometric and conformational isomers, including cis- or trans-conformations. The compounds may also exist in one or more tautomeric forms, including both single tautomers and mixtures of tautomers. The term "isomer" is intended to encompass all isomeric forms of the compounds of the present disclosure, including tautomers of the compounds. The compounds of the present disclosure may also exist in open-chain or cyclized forms. In some cases, one or more of the cyclized forms may result from loss of water. The specific composition of the open-chain and cyclized forms may depend on how the compound is isolated, stored or administered. For example, the compound may exist primarily in open-chain form under acidic conditions, but may cyclize under neutral conditions. All forms are included in the present disclosure.

[0046] Some compounds described herein may have asymmetric centers and therefore may exist in various enantiomeric and diastereomeric forms. Compounds described herein may be in the form of optical isomers or diastereomers. Thus, the present disclosure encompasses the compounds described herein in the form of optical isomers, diastereoisomers, and mixtures thereof (including racemic mixtures) and their use. Optical isomers of compounds of the present disclosure can be obtained by known techniques, such as asymmetric synthesis, chiral chromatography, simulated moving bed technique, or chemical separation of stereoisomers by using optically active resolving agents.

[0047] Unless otherwise indicated, the term "stereoisomer" means one stereoisomer of a compound that is substantially free of other stereoisomers of that compound. Thus, a stereoisomerically pure compound having one chiral center is substantially free of the opposite enantiomer of the compound. A stereoisomerically pure compound having two chiral centers is substantially free of other diastereomers of the compound. A typical stereoisomerically pure compound contains more than about 80% by weight of one stereoisomer of the compound and less than about 20% by weight of other stereoisomers of the compound, for example, more than about 90% by weight of one stereoisomer of the compound and less than about 10% by weight of other stereoisomers of the compound, or more than about 95% by weight of one stereoisomer of the compound and less than about 5% by weight of other stereoisomers of the compound, or more than about 97% by weight of one stereoisomer of the compound and less than about 3% by weight of other stereoisomers of the compound, or more than about 99% by weight of one stereoisomer of the compound and less than about 1% by weight of other stereoisomers of the compound. The above stereoisomers may be considered as compositions comprising the two stereoisomers present in the respective weight percentages set forth herein.

[0048] In the event of a discrepancy between a depicted structure and a name given to that structure, the depicted structure will control. Furthermore, if the stereochemistry of a structure or part of a structure is not shown, for example, with bold or dashed lines, the structure or part of a structure should be interpreted as embracing all stereoisomers of it. However, in some cases where multiple chiral centers are present, structures and names can be depicted as single enantiomers to help explain the relative stereochemistry. Those skilled in the art of organic synthesis will know whether compounds are prepared as single enantiomers from the methods used to prepare them.

[0049] As used herein, unless otherwise specified, the term "compound" is inclusive in that it encompasses a compound or its pharma- ceutically acceptable salts, stereoisomers, and / or tautomers. Thus, for example, a compound of the present disclosure includes a pharma- ceutically acceptable salt of a tautomer of the compound.

[0050] As used herein, a "pharmaceutical acceptable salt" refers to a pharmaceutical acceptable organic or inorganic acid or base salt of the compounds described herein. Representative pharmaceutical acceptable salts include, for example, alkali metal salts, alkaline earth salts, ammonium salts, water-soluble and water-insoluble salts, such as acetate, amsonate (4,4-diaminostilbene-2,2-disulfonate), benzenesulfonate, benzonate, bicarbonate, bisulfate, bitartrate, borate, bromide, butyrate, calcium, calcium edetate, camsylate, carbonate, chloride, citrate, clavulariate, and the like. clavulariate, dihydrochloride, edetate, edisylate, estolate, esylate, fiunarate, gluceptate, gluconate, glutamate, glycollylarsanilate, hexafluorophosphate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, Isothionate, lactate, lactobionate, laurate, malate, maleate, mandelate, mesylate, methyl bromide, methyl nitrate, methyl sulfate, mucate, napsylate, nitrate, N-methylglucamine ammonium salt, 3-hydroxy-2-naphthoate, oleate, oxalate, palmitate, pamoate (1,1-methene-bis-2-hydroxy-3-naphthoate, einbonate ate), pantothenate, phosphate / diphosphate, picrate, polygalacturonate, propionate, p-toluenesulfonate, salicylate, stearate, subacetate, succinate, sulfate, sulfosaliculate, suramate, tannate, tartrate, teoclate, tosylate, triethiodide and valerate. Pharmaceutically acceptable salts can have multiple charged atoms in their structure. In this example, the pharma- ceutically acceptable salts can have multiple counterions.Hence, a pharma- ceutically acceptable salt can have one or more charged atoms and / or one or more counter ion.

[0051] The terms "treat," "treating," and "treatment" refer to the amelioration or eradication of a disease or symptoms associated with a disease. In certain embodiments, such terms refer to minimizing the spread or worsening of a disease as a result of administration of one or more prophylactic or therapeutic agents to a patient with such a disease.

[0052] The terms "prevent," "preventing," and "prevention" refer to the prevention of the onset, recurrence, or spread of disease in a patient resulting from the administration of a prophylactic or therapeutic agent.

[0053] The term "effective amount" refers to an amount of a compound or other active ingredient described herein that is sufficient to provide a therapeutic or preventive benefit in the treatment or prevention of a disease, or to delay or minimize symptoms associated with a disease. Furthermore, a therapeutically effective amount in reference to a compound described herein refers to an amount of a therapeutic agent, alone or in combination with other therapies, that provides a therapeutic benefit in the treatment or prevention of a disease. When used in reference to a compound described herein, this term can include an amount that improves overall treatment, reduces or avoids symptoms or causes of a disease, or enhances or is synergistic with the therapeutic effectiveness of another therapeutic agent.

[0054] A "patient" or "subject" includes animals such as humans, cows, horses, sheep, lambs, pigs, chickens, turkeys, quail, cats, dogs, mice, rats, rabbits, or guinea pigs. According to some embodiments, the animals are mammals, such as non-primates and primates (e.g., monkeys and humans). In one embodiment, the patient is a human, such as a human infant, child, adolescent, or adult. In this disclosure, the terms "patient" and "subject" are used interchangeably.

[0055] By "inhibitor" is meant a compound that prevents or reduces the expression, catalytic activity, and / or localization (ie, local concentration) of DPP4.

[0056] compound As generally described above, the present disclosure provides compounds of formula (I) or a pharma- ceutically acceptable salt thereof. [ka]

[0057] In formula (I), each --- represents a single bond which, if present, forms a fused cyclopropyl ring.

[0058] L 1A is -NHCH2- or CH(NH2)-.

[0059] X 1 is selected from -O-, -S-, -S(O)-, S(O)2- and -NH-. 1B is C2-C 12 -alkyl, wherein one or more -CH2- are independently replaced by a moiety selected from -O-, -C(O)-, and -NH-. 1 H, C6-C 10 -aryl and 5-10 membered heteroaryl, wherein 1-4 heteroaryl members are independently selected from N, O and S.

[0060] The subscript m1 is Z 1 is H, then 0; Z 1 is an integer that is 1 if is other than H. The subscript n1 is an integer selected from 0, 1, 2, and 3.

[0061] R 1 , H, C1-C 10 -Alkyl and -C1-C 10 -Alkyl-(C6-C 10 -aryl), optionally substituted with 1 to 6 -OH.

[0062] R 2 is C1-C substituted with 1 to 6 -OH. 10 -alkyl. In various embodiments, R 2 is substituted with 1, 2, 3, 4, 5 or 6 -OH. 2 are shown below, along with examples of some of their diastereomers. [ka]

[0063] In various embodiments, L 1A In another embodiment, L 1A is -CH(NH2)-.

[0064] In some embodiments, X 1 is O.

[0065] In a further embodiment, Z 1 is H. In other embodiments, Z 1 is C6-C 10 -aryl or 5-10 membered heteroaryl, where 1-4 heteroaryl members are independently selected from N, O and S. According to one embodiment, C6-C 10 An example of -aryl is phenyl. An example of heteroaryl, according to another embodiment, is triazolyl.

[0066] In various embodiments, optionally in combination with any other embodiments described herein, n1 is 1 or 2.

[0067] In a further embodiment, optionally in combination with any other embodiment described herein, R 1 is C1-C optionally substituted with 1 to 6 -OH 10 -alkyl. For example, in an exemplary embodiment, R 1 is C1-C6-alkyl.

[0068] In yet an additional embodiment, optionally in combination with any other embodiment described herein, R 2 is a C2-C6-alkyl substituted with 1 to 5 -OH or R 2 is a C2-C6-alkyl substituted with 3 to 5 -OH. In an exemplary embodiment, R 2 is as follows: [ka]

[0069] A further embodiment provides a compound of formula (I), wherein: Z 1 is C6-C 10 -aryl or 5-10 membered heteroaryl, wherein 1-4 heteroaryl members are independently selected from N, O and S; n1 is 1 or 2; R 1 is C1-C 10 - alkyl; R 2 is a C2-C6-alkyl substituted with 1 to 5 -OH.

[0070] In some embodiments, the optional single bonds represented by "---" are absent. In other embodiments, they are present to form a fused cyclopropyl ring. These embodiments are each illustrated by the following structures: [ka]

[0071] As described herein, the moiety L 1B wherein one or more -CH2- groups may be independently replaced by -O-, -C(O)-, and -NH-; 12In some embodiments, 1, 2, 3, 4 or 5 -CH2- groups are replaced. It is understood that, according to chemical principles, substitutions will only form stable compounds, such as when substitution occurs at adjacent -CH2- groups. L 1B Examples include: [ka]

[0072] In other embodiments, the compound is of formula (II) or a pharma- ceutically acceptable salt thereof: [ka]

[0073] In formula (II), W is CH or N.

[0074] The subscript o is an integer selected from 1, 2, and 3.

[0075] R 3 is C1-C6-alkyl, C1-C6-hydroxyalkyl and -(CH2CH2O) x H, where x is an integer selected from 1, 2, 3, 4 and 5. 4 is C2-C8-alkynyl.

[0076] R 5a , R 5b , R 5c and R 5d is H, C1-C6-alkyl, halo, -NR A R B (R A and R B H and C1-C 10 -alkyl), -C(O)OH, -B(OH), -C(O)NR A R B , -C(O)OR A and -C(O)-L 2 -Z 2 -[(CH2)n2 -NR 6 R 7 ] m2 are independently selected from L 2 is C2-C 12 -alkyl, wherein one or more -CH2- are independently replaced by a moiety selected from -O-, -C(O)-, and -NH-. 2 H, C6-C 10 -aryl and 5-10 membered heteroaryl, where 1-4 heteroaryl members are independently selected from N, O and S. 5a , R 5b , R 5c and R 5d At least one of is other than H.

[0077] In addition, when W is CH, R 5a and R 5d is not selected from -C(O)OH, -C(O)OMe, and -C(O)OEt.

[0078] R 6 , H, C1-C 10 -Alkyl and -C1-C 10 -Alkyl-(C6-C 10 -aryl), optionally substituted with 1 to 6 -OH. 7 is C1-C substituted with 1 to 6 -OH. 10 -alkyl. In various embodiments, R 7 is substituted with 1, 2, 3, 4, 5 or 6 -OH. 7 are shown below, along with examples of some of their diastereomers: [ka]

[0079] The subscript m2 is Z 1 is H, then 0; Z 1 is an integer that is 1 if is other than H. The subscript n2 is an integer selected from 0, 1, 2, and 3.

[0080] In embodiments, W is CH. In other embodiments, W is N.

[0081] In various embodiments, R 5a , R 5b , R 5c and R 5d is H. In some embodiments, one, two, or three of R 5b and R 5d Each of R is H. 5c is halo, e.g., chloro. In yet other embodiments, R 5a is -C(O)OH or -C(O)OR A In a further embodiment, R 5a is -C(O)-L 2 -Z 2 -[(CH2) n2 -NR 6 R 7 ] m2 All of these embodiments and combinations thereof are contemplated.

[0082] R 5a , R 5b , R 5c and R 5d One of the following, for example R 5a -C(O)-L 2 -Z 2 -[(CH2) n2 -NR 6 R 7 ] m2 In relation to the embodiment where 2 is provided as phenyl or triazolyl. In an exemplary embodiment, Z 2 is triazolyl.

[0083] In an additional embodiment, optionally in combination with any other embodiment described herein, n2 is 1 or 2.

[0084] In yet a further embodiment, optionally in combination with any other embodiment described herein, R 6 is C1-C optionally substituted with 1 to 6 -OH 10 -alkyl. In one embodiment, R 6 is C1-C6-alkyl.

[0085] In an additional embodiment, optionally in combination with any other embodiment described herein, R 7 is a C2-C6-alkyl substituted with 1-5 -OH or 3-5 -OH. For example, in an exemplary embodiment, R 7 is: [ka]

[0086] In yet a further embodiment, the disclosure provides a compound of formula (II), wherein: R 5a , R 5b , R 5c and R 5d One of them is -C(O)-L 2 -Z 2 -[(CH2) n2 -NR 6 R 7 ] m2 and; Z 2 is C6-C 10 -aryl or 5-10 membered heteroaryl, wherein 1-4 heteroaryl members are independently selected from N, O and S; n2 is 1 or 2; R 6 is C1-C 10 - alkyl; R 7 is a C2-C6-alkyl substituted with 1 to 5 -OH.

[0087] As described herein, in various embodiments, the moiety L 2wherein one or more -CH2- groups may be independently replaced by -O-, -C(O)-, and -NH-; 12 In some embodiments, 1, 2, 3, 4 or 5 -CH2- groups are replaced. It is understood that, according to chemical principles, substitutions will only form stable compounds, such as when substitution occurs at adjacent -CH2- groups. L 2 Examples include: [ka]

[0088] In various embodiments, o is 1 or 2. In certain embodiments, o is 1.

[0089] Optionally in combination with any other embodiment, Z 2 But, C6-C 10 -aryl or 5-10 membered heteroaryl, where 1-4 heteroaryl members are independently selected from N, O and S. In one embodiment, Z 2 C6-C such as phenyl 10 -aryl.

[0090] In a further embodiment, the compound is of formula (III) or a pharma- ceutically acceptable salt thereof: [ka]

[0091] In formula (III), X 3 is -O- or -NH-. 3 is a bond or C2-C 12 -alkyl, wherein one or more -CH2- are independently replaced by a moiety selected from -O-, -C(O)-, and -NH-. 3 H, -N3, C6-C 10-aryl, 5-10 membered heteroaryl, where 1-4 heteroaryl members are independently selected from N, O and S, and (3-14 membered heterocycloalkyl, where 1-4 ring members are independently selected from N, O and S). Heteroaryl and heterocycloalkyl are optionally substituted with 1-6 substituents selected from the group consisting of halo, NO, OH, CN and C-C-haloalkyl.

[0092] The subscript m3 is Z 3 is H or -N3, then it is 0, and Z 3 is other than H or -N3. The subscript n3 is an integer selected from 0, 1, 2, and 3.

[0093] R 8 , H, C1-C 10 -Alkyl or -C1-C 10 -Alkyl-(C6-C 10 -aryl), optionally substituted with 1 to 6 -OH. 9 is C1-C substituted with 1 to 6 -OH. 10 -alkyl. R 10 is C1-C6-haloalkyl. 11 is independently selected from H, C1-C6-alkyl and halo.

[0094] The subscript o3 is an integer selected from 0, 1, 2, and 3. The subscript p3 is an integer selected from 0, 1, 2, and 3. The subscript q3 is an integer selected from 0, 1, 2, and 3.

[0095] In some embodiments, X 3 is O. In another embodiment, X 3 is -NH-.

[0096] As described herein, the moiety L 3 wherein one or more -CH2- groups may be independently replaced by -O-, -C(O)-, and -NH-;12 In some embodiments, the moiety L 3 is C5-C 12 -Alkyl, C5-C 10 -Alkyl, C7-C 12 -Alkyl, C8-C 12 -Alkyl or C9-C 12 -alkyl. In some embodiments, optionally in combination with any of the embodiments described herein, 1, 2, 3, 4 or 5 -CH2- groups are replaced. It should be understood that, according to chemical principles, the replacement will only form stable compounds, such as when the replacement occurs at adjacent -CH2- groups. 3 Examples include: [ka]

[0097] In various embodiments, Z 3 is -N3. Thus, in these embodiments, m3 is 0.

[0098] In another embodiment, Z 3 is C6-C 10 -aryl or 5-10 membered heteroaryl, where 1-4 heteroaryl members are independently selected from N, O and S. Exemplary Z 3 is phenyl or triazolyl.

[0099] In some embodiments, optionally in combination with any other embodiments described herein, n3 is 1 or 2.

[0100] In a further embodiment, optionally in combination with any other embodiment described herein, R 8 is C1-C optionally substituted with 1 to 6 -OH 10 -alkyl. In an exemplary embodiment, R 8 is C1-C6-alkyl.

[0101] In an additional embodiment, optionally in combination with any other embodiment described herein, R 9 is a C2-C6-alkyl substituted with 1 to 5 -OH, for example 3 to 5 -OH. In an exemplary embodiment, R 9 is as follows: [ka]

[0102] The present disclosure provides a compound of formula (III), wherein: Z 3 is C6-C 10 -aryl or 5-10 membered heteroaryl, wherein 1-4 heteroaryl members are independently selected from N, O and S; n3 is 1 or 2; R 8 is C1-C 10 - alkyl; R 9 is a C2-C6-alkyl substituted with 1 to 5 -OH.

[0103] Notwithstanding the general definitions set forth herein, it is understood that a compound does not include any of the following compounds: [Table 2] TIFF2024504328000021.tif210135

[0104] In still further embodiments, the present disclosure provides certain compounds, or pharma- ceutically acceptable salts thereof, as set forth in Tables 1 to 4 below. [Table 3] TIFF2024504328000023.tif183164TIFF2024504328000024.tif160163 [Table 4] [Table 5] TIFF2024504328000027.tif70163

[0105] Pharmaceutical Compositions The present disclosure also provides pharmaceutical compositions comprising a therapeutically effective amount of one or more compounds described herein, or pharma- ceutically acceptable salts, stereoisomers, and / or tautomers thereof, in admixture with a pharma- ceutically acceptable carrier. In some embodiments, the compositions further contain one or more additional therapeutic agents, pharma- ceutically acceptable excipients, diluents, adjuvants, stabilizers, emulsifiers, preservatives, colorants, buffers, flavoring agents, in accordance with accepted practices of pharmaceutical compounding.

[0106] In one embodiment, the pharmaceutical composition comprises a compound selected from the compounds shown in any table disclosed herein or a pharma- ceutically acceptable salt, stereoisomer and / or tautomer thereof, and a pharma- ceutically acceptable carrier.

[0107] The pharmaceutical compositions of the present disclosure are formulated, dosed, and administered in a manner consistent with good medical practice. Factors to be considered in this regard include the particular disorder being treated, the particular subject being treated, the clinical condition of the subject, the cause of the disorder, the site of delivery of the agent, the method of administration, the administration schedule, and other factors known to medical practitioners.

[0108] The "therapeutically effective amount" of the compound or its pharma- ceutically acceptable salt, stereoisomer and / or tautomer administered will depend on such considerations and is the minimum amount necessary to restore AEC2 cell proliferation or inhibit DPP4, or both. Such an amount may be less than an amount that is toxic to normal cells, or to the subject as a whole. In general, the initial therapeutically effective amount of the compound of the present disclosure (or its pharma- ceutically acceptable salt, stereoisomer or tautomer) administered will range from about 0.01 to about 200 mg per kg of patient body weight or from about 0.1 to about 20 mg per day, with a typical initial range being from about 0.3 to about 15 mg / kg per day. Oral unit dosage forms, such as tablets and capsules, may contain from about 0.1 mg to about 1000 mg of the compound of the present disclosure (or its pharma- ceutically acceptable salt, stereoisomer or tautomer). In another embodiment, such dosage forms contain about 50 mg to about 500 mg of the disclosed compound (or a pharma- ceutically acceptable salt, stereoisomer, or tautomer). In yet another embodiment, such dosage forms contain about 25 mg to about 200 mg of the disclosed compound (or a pharma- ceutically acceptable salt, stereoisomer, or tautomer). In yet another embodiment, such dosage forms contain about 10 mg to about 100 mg of the disclosed compound (or a pharma- ceutically acceptable salt, stereoisomer, or tautomer). In a further embodiment, such dosage forms contain about 5 mg to about 50 mg of the disclosed compound (or a pharma- ceutically acceptable salt, stereoisomer, or tautomer). In any of the foregoing embodiments, the dosage forms may be administered once daily or twice daily.

[0109] The compositions of the present disclosure can be administered orally, topically, parenterally, by inhalation or spray, or rectally in dosage unit formulations. The term parenteral as used herein includes subcutaneous injections, intravenous, intramuscular, intrasternal injection or infusion techniques.

[0110] Suitable oral compositions described herein include, but are not limited to, tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, syrups or elixirs.

[0111] In another aspect, also included is a pharmaceutical composition suitable for single unit dosage comprising a compound of the present disclosure or a pharma- ceutically acceptable stereoisomer, salt, or tautomer thereof and a pharma- ceutically acceptable carrier.

[0112] The composition of the present disclosure suitable for oral use can be prepared according to any method known in the art for the manufacture of pharmaceutical compositions.For example, the liquid formulation of the compound of the present disclosure contains one or more agents selected from the group consisting of sweeteners, flavoring agents, coloring agents and preservatives, in order to provide the medicamentous preparation of the compound.

[0113] For tablet composition, the compound of the present disclosure mixed with non-toxic pharma- ceutically acceptable excipients is used to manufacture tablets.Examples of such excipients include, but are not limited to, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents such as cornstarch or alginic acid; binders such as starch, gelatin or acacia, and lubricants such as magnesium stearate, stearic acid or talc.Tablets can be uncoated or coated by known coating techniques to delay disintegration and absorption in the gastrointestinal tract, thereby providing sustained therapeutic action over a desired period of time.For example, time-delay materials such as glyceryl monostearate or glyceryl distearate can be used.

[0114] Formulations for oral use may also be presented as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent, such as calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules in which the active ingredient is mixed with water or an oil medium, such as peanut oil, liquid paraffin or olive oil.

[0115] In the case of aqueous suspension, the compound of the present disclosure is mixed with suitable excipients to maintain a stable suspension.Examples of such excipients include, but are not limited to, sodium carboxymethylcellulose, methylcellulose, hydropropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum arabic.

[0116] Oral suspensions may also contain dispersing or wetting agents, such as natural phosphatides, e.g., lecithin, or condensation products of alkylene oxides with fatty acids, e.g., polyoxyethylene stearate, or condensation products of ethylene oxide with long-chain aliphatic alcohols, e.g., heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitols, e.g., polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, e.g., polyethylene sorbitan monooleate. Aqueous suspensions may also contain one or more preservatives, e.g., ethyl or n-propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents, e.g., sucrose or saccharin.

[0117] Oily suspensions can be formulated by suspending the compounds of the present disclosure in a vegetable oil, such as arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil, such as liquid paraffin. The oily suspensions may contain a thickening agent, such as beeswax, hard paraffin or cetyl alcohol.

[0118] Sweetening agents such as those set forth above, and flavoring agents may be added to provide a palatable oral preparation.These compositions may be preserved by the addition of an antioxidant such as ascorbic acid.

[0119] Dispersible powders and granules suitable for preparing aqueous suspension by adding water provide the compound of the present disclosure mixed with a dispersing or wetting agent, a suspending agent and one or more preservatives.Suitable dispersing or wetting agents and suspending agents are exemplified by those already mentioned above.Additional excipients, such as sweeteners, flavorings and colorings, can also be present.

[0120] The pharmaceutical composition of the present disclosure may be in the form of an oil-in-water emulsion.The oil phase may be vegetable oil, such as olive oil or peanut oil, or mineral oil, such as liquid paraffin, or mixtures thereof.Suitable emulsifiers may be natural gums, such as gum arabic or gum tragacanth, natural phosphatides, such as soybean, lecithin, and esters or partial esters derived from fatty acids and hexitols, anhydrides, such as sorbitan monooleate, and the condensation reaction products of the partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate.Emulsions may also contain sweeteners and flavoring agents.

[0121] Syrups and elixirs may be formulated with sweetening agents, for example, glycerol, propylene glycol, sorbitol or sucrose. Such formulations may also contain a demulcent, a preservative, and flavoring and coloring agents. The pharmaceutical compositions may be in the form of a sterile injectable, aqueous or oleaginous suspension. The suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents, as described above. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be used are water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any mild fixed oil may be employed, including synthetic mono- or diglycerides. In addition, fatty acids, such as oleic acid, are used in the preparation of injectables.

[0122] The compound of the present disclosure can also be administered in the form of suppositories for rectal administration.These compositions can be prepared by mixing the compound with suitable non-irritating excipients that are solid at normal temperature but liquid at rectal temperature, and therefore melt in rectum to release the compound.Such materials are cocoa butter and polyethylene glycol.

[0123] The composition for parenteral administration is administered in sterile medium.Depending on the vehicle used and the concentration of the compound in the preparation, parenteral preparation can be either a suspension or a solution containing dissolved compound.Adjuvants such as local anesthetics, preservatives and buffers can also be added to parenteral composition.

[0124] method The compounds of the present disclosure are useful as inhibitors of DPP4. They are further useful for selectively promoting proliferation of AEC2 cells without affecting the activation or proliferation of myofibroblasts, which is undesirable in most disease situations. In various embodiments, the compounds promote alveolar repair as a treatment for diseases whose etiology originates from epithelial degeneration and maladaptive remodeling. Exemplary indications include, but are not limited to, idiopathic pulmonary fibrosis (IPF), acute respiratory distress syndrome (ARDS) and infantile respiratory distress syndrome (IRDS).

[0125] The advantage of the compounds of the present disclosure is their ability to regulate the regenerative capacity of AEC2 cells.In contrast to this characteristic, the drugs approved for IPF, for example, inhibit the activation and proliferation of pulmonary fibroblasts and myofibroblasts, which are the source of scar tissue in diseased lungs.In contrast, the compounds of the present disclosure promote alveolar repair by directly targeting the source of disease in IPF, which is the ineffective self-renewal of damaged AEC2 cells.Therefore, the AEC2 targeting compounds of the present disclosure provide additional disease-modifying effects as single agents or as combination therapy with approved IPF drugs (e.g., pirfenidone).

[0126] Thus, the present disclosure provides, in various embodiments, a method for selectively increasing proliferation of cuboidal alveolar type 2 (AEC2) cells in a subject in need thereof, or for restoring decreased proliferation of AEC2 cells in a subject in need thereof, comprising administering to the subject a compound or pharma- ceutically acceptable salt disclosed herein.

[0127] Another embodiment also provides a method of inhibiting dipeptidyl peptidase IV (DPP4) in a subject in need thereof, comprising administering to the subject a compound or a pharma- ceutically acceptable salt disclosed herein.

[0128] In various embodiments, the present disclosure provides a method for treating a subject suffering from a disease or condition whose etiology is derived from epithelial degeneration, maladaptive remodeling, and / or ineffective self-renewal of damaged AEC2 cells. In some embodiments, the disease is a pulmonary disease or condition.In further embodiments, the disease or condition includes idiopathic pulmonary fibrosis (IPF), acute respiratory distress syndrome (ARDS), chronic obstructive pulmonary disease (COPD), emphysema, silicosis, asbestosis, pneumoconiosis, aluminosis, bauxite fibrosis, beryllium poisoning, siderosis, stannosis, talc, Labrador lung (mixed pneumoconiosis), sarcoidosis, hypersensitivity pneumonitis (HP) / extrinsic allergic alveolitis (EAA), chronic bronchitis, desquamative interstitial pneumonia (DIP), respiratory bronchiolitis interstitial lung disease (RBILD), acute interstitial pneumonia (AIP), nonspecific interstitial pneumonia (NS). IP), Cryptopathic Organizing Pneumonia (COP = Cryptopathic BOOP), Secondary Organizing Pneumonia (BOOP), Lymphoid Interstitial Pneumonia (LIP), Idiopathic Interstitial Pneumonia: Unspecified Eosinophilic Lung Disease, Tuberculosis (TB), Pulmonary Edema, Interstitial Lung Disease, Bronchopulmonary Dysplasia (BPD), Coronavirus, COVID-19, Cryptopathic Organizing Pneumonia (COP), Cystic Fibrosis (CF), E-Cigarette or Vaping Use Associated Lung Injury (EVALI), Hantavirus Pulmonary Syndrome (HPS), Histoplasmosis, Influenza, Legionnaires' Disease, MAC Lung Disease, Alpha-1 Antitrypsin Deficiency , aspergillosis, lymphangioleiomyoma (LAM), Middle East Respiratory Syndrome (MERS), nontuberculous mycobacterial lung disease (NTM), lung cancer, pulmonary embolism, Goodpasture's syndrome, idiopathic pulmonary hemophilia, cryptogenic alveolar hemorrhage syndrome, definite causative alveolar hemorrhage syndrome, sporadic pulmonary lymphangioleiomyoma (S-LAM), pulmonary lymphangioleiomyoma in tuberous sclerosis complex (TSC-LAM), pulmonary alveolar proteinosis, pulmonary amyloidosis, primary pulmonary lymphoma, primary ciliary dyskinesia (with or without situs inversus), rare causes of hypersensitivity pneumonitis (farmer's lung disease and pigeon breeder's) Interstitial lung disease in idiopathic inflammatory myopathies (polymyositis, dermatomyositis, antisynthetase syndrome), interstitial lung disease in Sjogren's syndrome, interstitial lung disease in mixed connective tissue disease (MCTD), interstitial lung disease in overlap syndromes, interstitial lung disease in undifferentiated connective tissue disease, and bronchiolitis obliterans (in non-transplant patients).

[0129] In other embodiments, the disease is selected from inflammatory diseases and other diseases and disorders.Diseases or disorders include infectious colitis, ulcerative colitis, Crohn's disease, ischemic colitis, radiation colitis, peptic ulcer, intestinal cancer, intestinal obstruction, rheumatoid arthritis, psoriatic arthritis, Hashimoto's thyroiditis, systemic lupus erythematosus, multiple sclerosis, Graves' disease, type 1 diabetes, psoriasis, ankylosing spondylitis, scleroderma, myositis, gout, antiphospholipid syndrome (APS), vasculitis, dilated cardiomyopathy, hypertrophic cardiomyopathy, restrictive cardiomyopathy, left heart failure, right heart failure, systolic heart failure, dilated heart failure (heart failure with preserved ejection fraction), atrial septal defect, atrioventricular septal defect, aortic Stenosis, Double Outlet Right Ventricle, d Transposition of the Great Arteries, Ebstein Anomaly, Hypoplastic Left Heart Syndrome, Disconnected Aortic Artery, Pulmonary Atresia, Single Ventricle, Tetralogy of Fallot, Total Anomalous Pulmonary Venous Return, Tricuspid Atresia, Truncus Arteriosus, Ventricular Septal Defect, Polycystic Kidney Disease, Diabetes Insipidus, Goodpasture's Disease, IgA Vasculitis, IgA Nephropathy, Lupus Nephritis, Adult Nephrotic Syndrome, Childhood Nephrotic Syndrome, Hemolytic Uremic Syndrome, Medullary Spongiosis, Renal Dysplasia, Renal Artery Stenosis, Renovascular Hypertension, Renal Tubular Acidosis, Alport Syndrome, Wenger's Granulomatosis, Alagille Syndrome, Cystic Kidney Disease, Glosis, Fabry disease, focal segmental glomerulosclerosis (FSGS), glomerulonephritis, aHUS (atypical hemolytic uremic syndrome), hemolytic uremic syndrome (HUS), Henoch-Schönlein purpura, IgA nephropathy (Berger's disease), interstitial nephritis, minimal change disease, nephrotic syndrome, thrombotic thrombocytopenic purpura (TTP), granulomatosis with polyangiitis (GPA), eczema, psoriasis, cellulitis, impetigo, atopic dermatitis, epidermolysis bullosa, lichen sclerosus, ichthyosis, vitiligo, acral peeling skin syndrome, Blau syndrome, primary cutaneous amyloidosis, skin Skin abscess, bedsore, blepharitis, furuncle disease, full or partial thickness burn, capillaritis, dermatitis, cellulitis, corneal abrasion, corneal erosion, xerosis, lichen planus, lichen simplex chronicus, venous ulcer (stasis ulcer), adult Still's disease, agammaglobulinemia, alopecia areata, autoimmune angioedema, autoimmune autonomic neuropathy, autoimmune encephalomyelitis, autoimmune hepatitis, autoimmune myocarditis, autoimmune oophoritis, autoimmune orchitis, autoimmune pancreatitis, autoimmune retinopathy, autoimmune urticaria, axonal and neuronal neuropathy (AMAN), Barrow's disease, bullous pemphigoid,Celiac disease, chronic relapsing multifocal osteomyelitis (CRMO), Churg-Strauss syndrome (CSS) or eosinophilic granulomatosis (EGPA), cicatricial pemphigoid, Cogan's syndrome, cold agglutinin disease, Coxsackie myocarditis, CREST syndrome, dermatitis herpetiformis, dermatomyositis, Devic's disease (neuromyelitis optica), discoid lupus, eosinophilic esophagitis (EoE), eosinophilic fasciitis, erythema nodosum, essential mixed cryoglobulinemia, giant cell arteritis (temporal arteritis), giant cell myocarditis, granulomatosis with polyangiitis, Guillain-Barré syndrome, Hashimoto's thyroiditis, Henoch-Schönlein Herpes gestationis or pemphigoid of gestationis (PG), hypogammaglobulinemia, IgG4-related sclerosing disease, immune thrombocytopenic purpura (ITP), inclusion body myositis (IBM), Lambert-Eaton syndrome, leukocytoclastic vasculitis, linear immunoglobulin A disease (LAD), microscopic polyangiitis (MPA), mixed connective tissue disease (MCTD), Mooren's ulcer, Mukka-Habermann syndrome, multifocal motor neuropathy (MMN) or MMNCB, multiple sclerosis, myasthenia gravis, myositis, narcolepsy, neonatal lupus, neuromyelitis optica, neutropenia, ocular cicatricial purpura Pemphigus, Optic neuritis, Relapsing rheumatism (PR), PANDAS, Paraneoplastic cerebellar degeneration (PCD), Paroxysmal nocturnal hemoglobinuria (PNH), Parry-Romberg disease, Parsplanitis (peripheral uveitis), Parsonage-Turner syndrome, Pemphigus, Peripheral neuropathy, Perivenous encephalomyelitis, Pernicious anemia (PA), POEMS syndrome, Polyarteritis nodosa, Polyglandular syndrome type I, II, III, Polymyalgia rheumatica, Polymyositis, Primary biliary cirrhosis, Primary sclerosing cholangitis, Progesterone dermatitis, Pure red cell aplasia (PRCA), Pyoderma gangrenosum, Raynaud's phenomenon, Reactive arthritis, Reflex sympathetic dystrophy, relapsing polychondritis, restless legs syndrome (RLS), retroperitoneal fibrosis, rheumatic fever, rheumatoid arthritis, sarcoidosis, Schmidt syndrome, scleritis, scleroderma, Sjogren's syndrome, sperm and testicular autoimmunity, stiff-person syndrome (SPS), subacute bacterial endocarditis (SBE), Susac syndrome, sympathetic ophthalmia (SO), Takayasu's arteritis, temporal arteritis / giant cell arteritis, thrombocytopenic purpura (TTP), thyroid eye disease (TED), Alagille syndrome, alcohol-related liver disease, autoimmune hepatitis, biliary atresia, cirrhosis,The diseases include those selected from lysosomal acid lipase deficiency (LAL-D), liver cysts, hepatoma, neonatal jaundice, nonalcoholic fatty liver disease, nonalcoholic steatohepatitis, primary biliary cholangitis (PBC), progressive familial intrahepatic cholestasis (PFIC), osteoporosis, Paget's disease, osteonecrosis, osteoarthritis, low bone mineral density, gout, fibrous dysplasia, Marfan syndrome, and osteogenesis imperfecta.

[0130] The reference numbers in the previous section are as follows:

[0131] [1] Hogan, BL, Barkauskas, CE, Chapman, HA, Epstein, JA, Jain, R., Hsia, CC, Niklason, L., Calle, E., Le, A., Randell, SH, Rock, J., Snitow, M., Krummel, M., Stripp, BR, Vu, T., White, ES, Whitsett, JAand. Morrisey, EE (2014) Repair and regeneration of the respiratory system: complexity, plasticity, and mechanisms of lung stem cell function, Cell Stem Cell 15, 123-138.

[0132] [2] Barkauskas,CE,Cronce,MJ,Rackley,CR,Bowie,EJ,Keene,DR,Stripp,BR,Randell,SH,Noble,PW,and Hogan,BL(2013)Type 2 alveolar cells are stem cells in adult lung,J Clin Invest 123,3025-3036. [3] Noble, PW, Barkauskas, CE, and Jiang, D. (2012) Pulmonary fibrosis: patterns and perpetrators, J Clin Invest 122, 2756-2762.

[0133] [4] Liang,J.,Zhang,Y.,Xie,T.,Liu,N.,Chen,H.,Geng,Y.,Kurkciyan,A.,Mena,J.M.,Stripp,B.R.,Jiang,D.,and Noble,P.W.(2016)Hyaluronan and TLR4 promote surfactant-protein-C-positive alveolar progenitor cell renewal and prevent severe pulmonary fibrosis in mice,Nat Med 22,1285-1293。

[0134] [5] Thompson,B.T.,Chambers,R.C.,and Liu,K.D.(2017)Acute Respiratory Distress Syndrome,N Engl J Med 377,1904-1905。

[0135] [6] Janes,J.,Young,M.E.,Chen,E.,Rogers,N.H.,Burgstaller-Muehlbacher,S.,Hughes,L.D.,Love,M.S.,Hull,M.V.,Kuhen,K.L.,Woods,A.K.,Joseph,S.B.,Petrassi,H.M.,McNamara,C.W.,Tremblay,M.S.,Su,A.I.,Schultz,P.G.,and Chatterjee,A.K.(2018)The ReFRAME library as a comprehensive drug repurposing library,and its application to the treatment of cryptosporidiosis,Proc Natl Acad Sci U S A 115,10750-10755. [7] Xu,J.,Wang,J.,He,M.,Han,H.,Xie,W.,Wang,H.,and Kong,H.(2018)Dipeptidyl peptidase IV(DPP-4)inhibition alleviates pulmonary arterial remodeling in experimental pulmonary hypertension,Lab Invest 98,1333-1346。

[0136] [8] Kawasaki,T.,Chen,W.,Htwe,Y.M.,Tatsumi,K.,and Dudek,S.M.(2018)DPP 4 inhibition by sitagliptin attenuates LPS-induced lung injury in mice,Am J Physiol Lung Cell Mol Physiol。

[0137] [9] Stone,M.L.,Sharma,A.K.,Zhao,Y.,Charles,E.J.,Huerter,M.E.,Johnston,W.F.,Kron,I.L.,Lynch,K.R.,and Laubach,V.E.(2015)Sphingosine-1-phosphate receptor 1 agonism attenuates lung ischemia-reperfusion injury,Am J Physiol Lung Cell Mol Physiol 308,L 1245-1252.

[10] Diab, KJ, Adamowicz, JJ, Kamocki, K., Rush, NI, Garrison, J., Gu, Y., Schweitzer, KS, Skobeleva, A., Rajashekhar, G., Hubbard, WC, Berdyshev, EV, and Petrache, I. (2010) Stimulation of sphingosine 1- signaling phosphate as a alveolar cell survival. Strategy in emphysema, Am J Respir Crit Care Med 181, 344-352.

[0138] Further embodiments of the present disclosure are described in the following non-limiting examples. All compounds of the present disclosure are made by procedures similar to those exemplified below.

[0139] [Example] intermediate compound Synthesis of (2R,3R,4R,5S)-6-(hexyl(prop-2-yn-1-yl)amino)hexane-1,2,3,4,5-pentaol (Int-1) [ka]

[0140] Step 1 (2R,3R,4R,5S)-6-(hexylamino)hexane-1,2,3,4,5-pentanol [ka]

[0141] To a stirred solution of hexane-1-amine (2.0 g, 19.76 mmol) and D-glucose (3.56 g, 19.76 mmol) in methanol (20 mL) was added Raney Ni (1.2 g) at room temperature. The reaction mixture was heated at 65° C. under H2 gas pressure (160 psi) for 16 h. After consumption of the starting material, the reaction mixture was filtered through a bed of Celite. The filtrate was concentrated under reduced pressure to give 4.5 g of (2R,3R,4R,5S)-6-(hexylamino)hexane-1,2,3,4,5-pentaol as an off-white solid [TLC system: MeOH:DCM (1:9); R f Value: 0.2].

[0142] Step 2 (2R,3R,4R,5S)-6-(hexyl(prop-2-yn-1-yl)amino)hexane-1,2,3,4,5-pentaol (Int-1): [ka]

[0143] To a stirred solution of (2R,3R,4R,5S)-6-(hexylamino)hexane-1,2,3,4,5-pentaol (1.0 g, 3.77 mmol) in THF (15 mL) was added 3-bromoprop-1-yne (80% solution in toluene, 0.84 mL, 5.65 mmol) and the reaction mixture was stirred at 80° C. for 16 h. After consumption of the starting material, the reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give 0.5 g of (2R,3R,4R,5S)-6-(hexyl(prop-2-yn-1-yl)amino)hexane-1,2,3,4,5-pentaol as a yellow gum. [TLC system: MeOH:DCM (1:9); R f Value: 0.3].

[0144] (1R,3S,5S)-3-((tert-butoxycarbonyl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl methanesulfonate (Int-2) [ka]

[0145] Step 1 tert-Butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-hydroxyadamantan-1-yl)carbamate: [ka]

[0146] A mixture of (2S)-1-((((1S,3R,5S)-3-hydroxyadamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (200 g, 659 mmol, 1.00 equiv.) and (Boc)O (165 g, 758 mmol, 1.15 equiv.) in THF (400 mL) and t-BuOH (400 mL) was degassed and purged with N three times, then the mixture was incubated at 70 °C under N atmosphere for 12 h. The mixture was stirred. The solution was concentrated under reduced pressure and MTBE (200 mL) was added. The mixture was filtered to give tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-hydroxyadamantan-1-yl)carbamate (239 g, 592 mmol, 89.8% yield, 99.7% purity) as a white solid. TLC system: PE:EA (0:1);R f :0.40.

[0147] Step 2 (1R,3S,5S)-3-((tert-butoxycarbonyl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl methanesulfonate (Int-2): [ka]

[0148] To a solution of compound tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-hydroxyadamantan-1-yl)carbamate (230 g, 570 mmol, 1.00 equiv) and N,N,N',N'-tetramethyl-1,6-hexanediamine (216 g, 1.25 mol, 2.20 equiv) in DCM (2.30 L) was cooled to 0-10 °C and methanesulfonyl chloride (97.9 g, 855 mmol, 1.50 equiv) was added at 0-10 °C. The reaction was stirred at 15-20 °C for 2 h. The reaction mixture was quenched by adding ice water (1.00 L) at 0 °C and extracted with EA (2.00 L). The combined organic layers were washed with 1.00 L of brine, dried over Na2SO4, filtered and concentrated under reduced pressure to give (1R,3S,5S)-3-((tert-butoxycarbonyl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl methanesulfonate (440 g, crude) as a pale yellow oil. TLC system: PE:EA (10:1); f :0.60) Synthesis of tert-butyl ((1S,3R,5S)-3-(2-(2-azidoethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamate (Int-3) [ka]

[0149] Step 1 tert-Butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-(2-hydroxyethoxy)ethoxy)adamantan-1-yl)carbamate: [ka]

[0150] To a stirred solution of (1R,3S,5S)-3-((tert-butoxycarbonyl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl methanesulfonate (3.58 g, 7.43 mmol) and 2,2'-oxybis(ethan-1-ol) (35.2 mL, 371.67 mmol) in acetonitrile (10 mL), molecular sieves 4 Å (4 g) were added and the mixture was stirred at 70° C. for 16 h. After completion, the reaction mixture was diluted with water (500 mL) and the resulting mixture was extracted with ethyl acetate (3×500 mL). The combined organic layers were dried over anhydrous sodium sulfate and evaporated to give the crude product. The crude product was then purified by silica gel column chromatography (230-400 mesh) using 3% methanol in dichloromethane to give tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-(2-hydroxyethoxy)ethoxy)adamantan-1-yl)carbamate (2.7 g) as a brown gum. TLC system: MeOH:DCM (1:9); R f :0.4 Step 2. 2-(2-(((1R,3S,5S)-3-((tert-butoxycarbonyl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl)oxo)ethoxy)ethyl methanesulfonate: [ka]

[0151] To a stirred solution of tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-(2-hydroxyethoxy)ethoxy)adamantan-1-yl)carbamate (2.7 g, 5.49 mmol) in dichloromethane (25 mL) was added triethylamine (1.67 g, 16.47 mmol) followed by methanesulfonyl chloride (1.26 g, 10.98 mmol) in dichloromethane (5 mL) at −10° C. The resulting mixture was stirred at −10° C. for 1 h. After completion, the reaction mixture was diluted with water (100 mL) and the resulting mixture was extracted with dichloromethane (2×300 mL). The combined organic layers were dried over anhydrous sodium sulfate and evaporated to give 2-(2-(((1R,3S,5S)-3-((tert-butoxycarbonyl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl)oxy)ethoxy)ethyl methanesulfonate (3.1 g) as a pale yellow foamy solid. TLC system: MeOH:DCM (0.5:9.5);R f : 0.4 (TLC eluted twice) Step 3 tert-Butyl ((1S,3R,5S)-3-(2-(2-azidoethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamate (Int-3): [ka]

[0152] To a stirred solution of 2-(2-(((1R,3S,5S)-3-((tert-butoxycarbonyl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl)oxy)ethoxy)ethyl methanesulfonate (3.1 g, 5.44 mmol) in N,N-dimethylformamide (10 mL) was added sodium azide (0.71 g, 10.88 mmol) and the reaction mixture was stirred at 70° C. for 16 h. After completion, the reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (3×300 mL). The combined organic layers were dried over anhydrous sodium sulfate and evaporated to give tert-butyl ((1S,3R,5S)-3-(2-(2-azidoethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamate (2.5 g) as a yellow gum. TLC system: MeOH:DCM (0.5:9.5); R f : 0.3 (TLC eluted twice) Synthesis of (2R,2'R,3R,3'R,4R,4'R,5S,5'S)-6,6'-(prop-2-yn-1-ylazanediyl)bis(hexane-1,2,3,4,5-pentanol) (Int-4) [ka]

[0153] To a mixture of (2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanal (2.0 g, 11.08 mmol), prop-2-yn-1-amine (0.3 g, 5.54 mmol) and acetic acid (2.0 mL) in methanol (16 mL) was added sodium cyanoborohydride (0.7 g, 11.08 mmol). The reaction mixture was stirred at 60° C. for 16 h. After completion (reaction monitored by LCMS), the reaction mixture was evaporated and the residue was washed with methanol (2×20 mL), diethyl ether (2×20 mL) and dried to give (2R,2′R,3R,3′R,4R,4′R,5S,5′S)-6,6′-(prop-2-yn-1-ylazanediyl)bis(hexane-1,2,3,4,5-pentanol) (1.3 g) as an off-white solid.

[0154] Synthesis of tert-butyl ((1S,3R,5S)-3-(2-(2-aminoethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamate (Int-5) [ka]

[0155] To a stirred solution of tert-butyl ((1S,3R,5S)-3-(2-(2-azidoethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamate (1.0 g, 1.94 mmol) in methanol (10 mL) was added triphenylphosphine (0.76 g, 2.9 mmol) and the reaction mixture was stirred at 70° C. for 16 h. After completion of the reaction, the reaction mixture was evaporated and the residue was purified by silica gel column chromatography (basified by triethylamine) using 6% methanol in dichloromethane to give tert-butyl ((1S,3R,5S)-3-(2-(2-aminoethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamate (0.28 g) as a pale yellow solid. TLC system: MeOH:DCM (1:9); Rf :0.2 Synthesis of tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-(2-hydroxyethoxy)ethoxy)adamantan-1-yl)carbamate (Int-6) [ka]

[0156] Step 1 tert-Butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-hydroxyadamantan-1-yl)carbamate: [ka]

[0157] To a stirred suspension of (2S)-1-(((1S,3R,5S)-3-hydroxyadamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (15 g, 49.44 mmol) in toluene (150 mL), triethylamine (13.78 mL, 98.88 mmol) and di-tert-butyl dicarbonate (17.0 mL, 74.16 mmol) were added and the reaction mixture was stirred at 100° C. for 16 h. After completion, the reaction mixture was evaporated to give a crude residue which was then purified by silica gel (230-400 mesh) column chromatography using 3% methanol in dichloromethane to give tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-hydroxyadamantan-1-yl)carbamate (17.0 g) as an off-white solid. TLC system: MeOH:DCM (0.5:9.5); R f : 0.2 (TLC eluted twice) Step 2 (1R,3S,5S)-3-((tert-butoxycarbonyl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl methanesulfonate: [ka]

[0158] To a stirred solution of tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-hydroxyadamantan-1-yl)carbamate (0.5 g, 1.24 mmol) in dichloromethane (10 mL) was added triethylamine (0.38 g, 3.72 mmol) followed by methanesulfonyl chloride (0.21 g, 1.86 mmol) in dichloromethane (2.5 mL) at −10° C. The reaction mixture was stirred at −10° C. for 30 minutes. After completion, the reaction mixture was diluted with water (10 mL) and the resulting mixture was extracted with dichloromethane (2×30 mL). The combined organic layers were dried over anhydrous sodium sulfate and evaporated to give (1R,3S,5S)-3-((tert-butoxycarbonyl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl methanesulfonate (0.59 g) as a pale yellow foamy solid. TLC system: MeOH:DCM (0.5:9.5); f : 0.3 (TLC eluted twice) Step 3 tert-Butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-(2-hydroxyethoxy)ethoxy)adamantan-1-yl)carbamate (Int-6): [ka]

[0159] To a stirred solution of (1R,3S,5S)-3-((tert-butoxycarbonyl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl methanesulfonate (3.58 g, 7.43 mmol) and 2,2'-oxybis(ethan-1-ol) (35.2 mL, 371.67 mmol) in acetonitrile (10 mL), molecular sieves 4 Å (4 g) were added and the mixture was stirred at 70° C. for 16 h. After completion, the reaction mixture was diluted with water (500 mL) and the resulting mixture was extracted with ethyl acetate (3×500 mL). The combined organic layers were dried over anhydrous sodium sulfate and evaporated to give the crude product. The crude product was then purified by silica gel column chromatography (230-400 mesh) using 3% methanol in dichloromethane to give tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-(2-hydroxyethoxy)ethoxy)adamantan-1-yl)carbamate (2.7 g) as a brown gum. TLC system: MeOH:DCM (1:9); R f :0.4 Synthesis of (2R,3R,4R,5S)-6-((4-(2-aminoethoxy)phenethyl)(hexyl)amino)hexane-1,2,3,4,5-pentaol (Int-7) [ka]

[0160] Step 1. 4-(2-(hexylamino)ethyl)phenol: [ka]

[0161] To a solution of 4-(2-aminoethyl)phenol (15 g, 109 mmol) and hexanal (10 g, 100 mmol) in EtOH (100 mL) was added AcOH (1 drop). The reaction mixture was stirred at 25 °C for 12 h. Then NaBH4 (2 g, 54 mmol) was added and the reaction mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated under pressure at 40 °C. The residue was diluted with water and extracted with EtOAc (200 mL x 2). The organic layer was washed with brine (50 mL) and then dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under vacuum to give the crude product. The crude product was purified by flash column chromatography eluting with (CH2Cl2 / MeOH, 0%-10%) to give 4-(2-(hexylamino)ethyl)phenol (6 g, 24% yield) as a yellow oil. ES MS M / Z=222 (M+1) Step 2 Benzylhexyl(4-hydroxyphenethyl)carbamate: [ka]

[0162] To a solution of 4-[2-(hexylamino)ethyl]phenol (6.2 g, 28 mmol) and DIEA (5.41 g, 42 mmol) in DCM (100 mL) was added a solution of CbzCl (4.29 g, 25 mmol) in DCM (10 mL) at 0 °C. The reaction mixture was stirred at 0 °C for 1 h. The mixture was then diluted with H2O (100 mL) and extracted with DCM (100 mL x 2). The DCM phase was dried over Na2SO4 and concentrated. The residue was purified by flash column chromatography eluting with (PE / EtOAc, 5%-20%) to give benzylhexyl(4-hydroxyphenethyl)carbamate (7.63 g, 76% yield) as a yellow oil. ES MS M / Z=356(M+1) Step 3 Benzyl (4-(2-(1,3-dioxoisoindolin-2-yl)ethoxy)phenethyl)(hexyl)carbamate: [ka]

[0163] To a mixture of benzyl hexyl (4-hydroxyphenethyl)carbamate (8.63 g, 24.3 mmol), 2-(2-hydroxyethyl)isoindole-1,3-dione (4.64 g, 24.3 mmol), PPh3 (12.73 g, 48.6 mol) in THF (50 mL) was added a solution of ADDP (12.24 g, 48.6 mmol) in THF (30 mL) at 25 °C under N2 atmosphere. The reaction mixture was stirred at 70 °C under N2 atmosphere for 12 h. The reaction mixture was concentrated under pressure at 40 °C. The residue was purified by flash column chromatography eluting with (PE / EtOAc, 10%-50%) to give benzyl (4-(2-(1,3-dioxoisoindolin-2-yl)ethoxy)phenethyl)(hexyl)carbamate (10.4 g, 81% yield) as a yellow oil. ES MS M / Z=529(M+1) Step 4. 2-(2-(4-(2-(hexylamino)ethyl)phenoxy)ethyl)isoindoline-1,3-dione: [ka]

[0164] To a solution of benzyl (4-(2-(1,3-dioxoisoindolin-2-yl)ethoxy)phenethyl)(hexyl)carbamate (4.86 g, 9.20 mmol) in MeCN (10 mL) was added TMSI (5 g, 35.7 mmol). The reaction mixture was stirred at 25 °C for 10 min. The reaction mixture was concentrated under pressure at 40 °C. The residue was purified by flash column chromatography eluting with (CHCl / MeOH, 5%-10%) to give 2-(2-(4-(2-(hexylamino)ethyl)phenoxy)ethyl)isoindoline-1,3-dione (2.56 g, 70% yield) as a yellow solid. ES MS M / Z=395(M+1). Step 5. 2-(2-(4-(2-(hexyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethyl)phenoxy)ethyl)isoindoline-1,3-dione: [ka]

[0165] To a solution of 2-(2-(4-(2-(hexylamino)ethyl)phenoxy)ethyl)isoindoline-1,3-dione (2.92 g, 7.41 mmol) and (2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanal (4.00 g, 22.23 mmol) in MeOH (20 mL) was added AcOH (10 drops). The reaction mixture was stirred at 25° C. for 1 h. NaBH3CN (1.86 g, 29.64 mmol) was then added and the reaction mixture was stirred at 50° C. for 12 h. To this reaction mixture, (2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanal (1.33 g, 14.82 mmol) was added every 12 hours until 2-(2-(4-(2-(hexylamino)ethyl)phenoxy)ethyl)isoindoline-1,3-dione disappeared (a total of about 4 equivalents of (2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanal was added to the reaction mixture, and the total reaction time was about 3 days). The reaction mixture was concentrated under pressure at 40°C. The residue was purified by flash column chromatography, eluting with (DCM / MeOH, 5%-10%) to give impure 2-(2-(4-(2-(hexyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethyl)phenoxy)ethyl)isoindoline-1,3-dione (4 g, 87% yield) as a colorless oil. ES MS M / Z=559(M+1). Step 6 (2R,3R,4R,5S)-6-((4-(2-aminoethoxy)phenethyl)(hexyl)amino)hexane-1,2,3,4,5-pentaol (Int-7): [ka]

[0166] To a solution of 2-(2-(4-(2-(hexyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethyl)phenoxy)ethyl)isoindoline-1,3-dione (3.5 g, 6.27 mmol) in EtOH (20 mL) was added hydrazine hydrate (10 mL). The reaction mixture was stirred at 80 °C for 4 h. The reaction mixture was concentrated under pressure at 40 °C. The residue was purified by flash column chromatography eluting with (MeOH / DCM, 20%-50%) to give (2R,3R,4R,5S)-6-((4-(2-aminoethoxy)phenethyl)(hexyl)amino)hexane-1,2,3,4,5-pentaol (2.5 g, 93% yield) as a colorless oil. ES MS M / Z=429(M+1) Synthesis of (2R,3R,4R,5S)-6-(prop-2-yn-1-ylamino)hexane-1,2,3,4,5-pentaol (Int-8) [ka]

[0167] To a mixture of (2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanal (1.0 g, 5.54 mmol), prop-2-yn-1-amine (0.61 g, 11.08 mmol) and acetic acid (1.0 mL) in methanol (20 mL) was added sodium cyanoborohydride (0.35 g, 5.54 mmol) at 0° C. The reaction mixture was stirred at room temperature for 16 h. After completion (reaction monitored by LCMS), the reaction mixture was evaporated and the residue was washed with diethyl ether (2×20 mL) and dried to give (2R,3R,4R,5S)-6-(prop-2-yn-1-ylamino)hexane-1,2,3,4,5-pentaol (1.0 g, crude) as an off-white solid.

[0168] Synthesis of tert-butyl ((1S)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-1-((1S,3R,5S)-3-(2-hydroxyethoxy)adamantan-1-yl)-2-oxoethyl)carbamate (Int-9) [ka]

[0169] Step 1 tert-Butyl ((1S)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-1-((1S,3R,5S)-3-hydroxyadamantan-1-yl)-2-oxoethyl)carbamate: [ka]

[0170] To a stirred suspension of (1S,3S,5S)-2-((2S)-2-amino-2-((1S,3R,5S)-3-hydroxyadamantan-1-yl)acetyl)-2-azabicyclo[3.1.0]hexane-3-carbonitrile (2 g, 6.34 mmol) in dichloromethane (40 mL) was added triethylamine (1.8 mL, 12.68 mmol) followed by di-tert-butyl dicarbonate (2.2 mL, 9.51 mmol) at 0° C. and the reaction mixture was stirred at room temperature for 3 h. After completion, the reaction mixture was evaporated to give a crude residue, which was then purified by silica gel (230-400 mesh) column chromatography using 3% methanol in dichloromethane to give tert-butyl ((1S)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-1-((1S,3R,5S)-3-hydroxyadamantan-1-yl)-2-oxoethyl)carbamate (2.5 g) as an off-white solid. TLC system: MeOH:DCM (0.5:9.5); R f :0.4 Step 2 (1R,3S,5S)-3-((S)-1-((tert-butoxycarbonyl)amino)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)adamantan-1-yl methanesulfonate: [ka]

[0171] To a stirred solution of tert-butyl ((1S)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-1-((1S,3R,5S)-3-hydroxyadamantan-1-yl)-2-oxoethyl)carbamate (2.5 g, 6.02 mmol) in dichloromethane (50 mL) was added triethylamine (2.5 mL, 18.06 mmol) followed by methanesulfonyl chloride (1.03 g, 9.03 mmol) in dichloromethane (2.5 mL) at -10°C and the mixture was stirred for 30 minutes. After completion, water (50 mL) was added to the reaction mixture and extracted with dichloromethane (2 x 100 mL). The combined organic layers were dried over anhydrous sodium sulfate and evaporated to give crude (1R,3S,5S)-3-((S)-1-((tert-butoxycarbonyl)amino)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)adamantan-1-yl methanesulfonate (3.2 g) as a white foamy solid. TLC system: 100% EtOAc; R f :0.4. Step 3 tert-Butyl ((1S)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-1-((1S,3R,5S)-3-(2-hydroxyethoxy)adamantan-1-yl)-2-oxoethyl)carbamate (Int-9): [ka]

[0172] To a stirred solution of (1R,3S,5S)-3-((S)-1-((tert-butoxycarbonyl)amino)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)adamantan-1-yl methanesulfonate (3.2 g, 6.48 mmol) and ethane-1,2-diol (18.2 mL, 324.13 mmol) in acetonitrile (30 mL) was added molecular sieves 4 Å (3.5 g) and the resulting mixture was stirred at 70° C. for 16 h. After completion, the reaction mixture was concentrated and the crude product was filtered through a Buchner funnel, diluted with water (100 mL) and extracted with ethyl acetate (3×100 mL). The combined organic layers were dried over anhydrous sodium sulfate and evaporated to give crude tert-butyl ((1S)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-1-((1S,3R,5S)-3-(2-hydroxyethoxy)adamantan-1-yl)-2-oxoethyl)carbamate (2.0 g) as an off-white foamy solid. TLC system: 100% EtOAc, R f :0.3 tert-Butyl ((1S)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-1-((1S,3R,5S)-3-(2-(2-hydroxyethoxy)ethoxy)adamantan-1-yl)-2-oxoethyl)carbamate (Int-9B) was prepared following the same route.

[0173] Synthesis of (2R,3R,4R,5S)-6-((4-(aminomethoxy)benzyl)(hexyl)amino)hexane-1,2,3,4,5-pentaol (Int-10) [ka]

[0174] To a suspension of N-hexyl-D-glucamine (840 mg, 1.05 equiv.) and potassium carbonate (594 mg, 1.5 equiv.) in methanol (8 mL) was added tert-butyl 4-(bromomethyl)benzylcarbamate (820 mg, 1.0 equiv.) portionwise at 0° C. After stirring at room temperature for 4 h, the reaction mixture was filtered through a pad of Celite with dichloromethane. The filtrate was then washed with water and concentrated in vacuo to give the crude product as a yellow oil, which was used in the next step without further purification. The above crude product (700 mg) was dissolved in 4 M HCl in dioxane (4.5 mL) and methanol (0.5 mL) at 0° C. After stirring at room temperature for 3 h, the volatiles were removed under reduced pressure to give (2R,3R,4R,5S)-6-((4-(aminomethyl)benzyl)(hexyl)amino)hexane-1,2,3,4,5-pentaol hydrochloride as a white powder, which was used in the next step without further purification.

[0175] Synthesis of 1-azido-2-(2-(2-bromoethoxy)ethoxy)ethane (Int-11): [ka]

[0176] Step 1. 2-(2-(2-hydroxyethoxy)ethoxy)ethyl 4-methylbenzenesulfonate: [ka]

[0177] To a stirred solution of compound 2,2'-(ethane-1,2-diylbis(oxy))bis(ethan-1-ol) (15 g, 99.88 mmol) in DCM (150 mL) was added silver oxide (27.7 g, 119.85 mmol) followed by potassium iodide (1.6 g, 9.98 mmol) at room temperature under nitrogen atmosphere. The resulting reaction mixture was stirred at room temperature for 10 min. p-TsCl (19 g, 99.88 mmol) was added and the resulting reaction mixture was stirred at room temperature for 16 h. The progress of the reaction was monitored by TLC. After completion of the starting material, the reaction mixture was concentrated under reduced pressure to obtain the crude compound. The crude compound was purified by column chromatography using silica gel (230-400 mesh) eluted with 10% EtOAC-petroleum ether to obtain 11 g of compound 5 as a colorless liquid. [TLC system: EtOAC:petroleum ether (2:8); R f Value: 0.5].

[0178] Step 2. 2-(2-(2-azidoethoxy)ethoxy)ethan-1-ol: [ka]

[0179] To a stirred solution of compound 2-(2-(2-hydroxyethoxy)ethoxy)ethyl 4-methylbenzenesulfonate (11 g, 36.14 mmol) in DMF (200 mL) was added sodium azide (4.7 g, 72.28 mmol) at room temperature. The resulting reaction mixture was heated at 120° C. for 6 h. The progress of the reaction was monitored by TLC. After completion of the starting material, the reaction mixture was concentrated under reduced pressure to obtain the crude compound. The crude compound was dissolved in diethyl ether (150 mL) and the solid precipitate was removed by filtration through a celite pad. The filtrate was collected and concentrated under reduced pressure to obtain the crude compound. The crude compound was purified by column chromatography using silica gel (230-400 mesh) eluted with 10% EtOAC-petroleum ether to obtain 5 g of compound 6 as an off-white solid. [TLC system: EtOAC:petroleum ether (3:7); R f Value: 0.3].

[0180] Step 3. 1-Azido-2-(2-(2-bromoethoxy)ethoxy)ethane (Int-11): [ka]

[0181] To a stirred solution of CBr4 (4.1 g, 12.56 mmol) in DCM (40 mL) was added Ph3P (3.3 g, 12.56 mmol). The reaction mixture was cooled to 0° C. and compound 2-(2-(2-azidoethoxy)ethoxy)ethan-1-ol (2 g, 11.42 mmol) was diluted with DCM (15 mL) and slowly added dropwise to the reaction mixture. The reaction mixture was stirred at room temperature for 8 hours. After completion of the starting material, the reaction mixture was quenched with water (20 mL) and extracted with DCM (2X50 mL). The combined organic layer was concentrated under reduced pressure to obtain the crude compound. The crude compound was purified by column chromatography using silica gel (230-400 mesh) eluted with 15% EtOAC-petroleum ether to obtain 1.2 g of Int-11 as a colorless liquid. [TLC system: EtOAC:petroleum ether (1:1); R f Value: 0.7].

[0182] Synthesis of tert-butyl (R)-(1-(7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-8-yl)piperidin-3-yl)carbamate (Int-13) [ka]

[0183] Step 1. 8-Bromo-7-(but-2-yn-1-yl)-3-methyl-3,7-dihydro-1H-purine-2,6-dione: [ka]

[0184] To a stirred solution of compound 8-bromo-3-methyl-3,7-dihydro-1H-purine-2,6-dione (10.0 g, 40.81 mmol) in DMF (100 mL), DIPEA (7.13 mL, 40.81 mmol) and 1-bromobut-2-yne (5.43 g, 40.81 mmol) were added portionwise at room temperature. The reaction mixture was stirred at room temperature for 16 hours. After completion of the starting material, the reaction mixture was quenched with cold water, and the precipitated solid was filtered off and dried under vacuum to give 9.3 g of compound 8-bromo-7-(but-2-yn-1-yl)-3-methyl-3,7-dihydro-1H-purine-2,6-dione as an off-white solid. [TLC system: MeOH:DCM (1:9); R f Value: 0.7].

[0185] Step 2 tert-Butyl (R)-(1-(7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-8-yl)piperidin-3-yl)carbamate (Int-13): [ka]

[0186] To a stirred solution of 8-bromo-7-(but-2-yn-1-yl)-3-methyl-3,7-dihydro-1H-purine-2,6-dione (4.5 g, 15.15 mmol) in DMF (45 mL) was added tert-butyl (R)-piperidin-3-ylcarbamate (3.33 g, 16.66 mmol) and K2CO3 (6.28 g, 45.45 mmol) and the reaction mixture was stirred at 90 °C for 16 h. After completion of starting material, the reaction mixture was quenched with cold water, and the precipitated solid was filtered off and dried under vacuum to give 5.0 g of tert-butyl (R)-(1-(7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-8-yl)piperidin-3-yl)carbamate as off-white solid. [TLC system: EtOAc:petroleum ether (1:1); Rf value: 0.3].

[0187] Synthesis of methyl (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-fluoronicotinate (Int-14) [ka]

[0188] Step 1 Methyl 6-fluoro-2-methylnicotinate: [ka]

[0189] To a stirred solution of 6-fluoro-2-methylnicotinic acid (3.0 g, 19.34 mmol) in THF (36 mL) and methanol (9 mL) was added 2M TMS diazomethane solution in toluene (12.57 mL, 25.14 mmol) at 0° C. and the reaction mixture was warmed to room temperature for 2 h. After completion of the reaction, the volatiles were removed under reduced pressure to obtain the crude compound. The crude compound was purified by column chromatography (silica gel 100-200 mesh, 0-10% EtOAc in petroleum ether used as eluent) to obtain compound methyl 6-fluoro-2-methylnicotinate 1.6 g as a colorless liquid product. [TLC system: EtOAc: petroleum ether (3:7); R f Value: 0.8].

[0190] Step 2 Methyl 2-(bromomethyl)-6-fluoronicotinate: [ka]

[0191] Methyl 6-fluoro-2-methylnicotinate To a stirred solution of methyl 6-fluoro-2-methylnicotinate (1.6 g, 9.46 mmol) in 1,2-dichloroethane (16 mL) was added N-bromosuccinimide (1.68 g, 9.46 mmol) followed by catalytic benzoyl peroxide (0.11 g, 0.47 mmol) in small portions. The reaction mixture was stirred at 70° C. for 16 hours. After completion of starting material, the solvent was evaporated under reduced pressure, quenched with water (100 mL) and the compound was extracted with dichloromethane (100 mL* 2). The combined organic layer was washed with brine solution (50 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain crude compound. The crude compound was purified by column chromatography (silica gel 230-400 mesh, 0-4% EtOAc in petroleum ether used as eluent) to give 1.2 g of compound methyl 2-(bromomethyl)-6-fluoronicotinate as a colorless liquid product. [TLC system: EtOAc: petroleum ether (1:9); R f Value: 0.7].

[0192] Step 3 Methyl (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-fluoronicotinate (Int-14): [ka]

[0193] Methyl 2-(bromomethyl)-6-fluoronicotinate To a stirred solution of methyl 2-(bromomethyl)-6-fluoronicotinate (0.6 g, 2.42 mmol) in DMF (6 mL), the compound tert-butyl (R)-(1-(7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-8-yl)piperidin-3-yl)carbamate (1.01 g, 2.42 mmol) and K2CO3 (0.33 g, 2.42 mmol) were added and the reaction mixture was stirred at 70° C. for 16 hours. After completion of the starting material, the reaction mixture was quenched with cold water, and the precipitated solid was filtered off and dried under vacuum to give 1 g of compound methyl (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-fluoronicotinate as an off-white solid. [TLC system: EtOAc:petroleum ether (1:1); R f Value: 0.4].

[0194] Synthesis of (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-(methylamino)nicotinic acid (Int-15) [ka]

[0195] Sep 1. Methyl (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-(methylamino)nicotinate: [ka]

[0196] To a stirred solution of the compound methyl (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-fluoronicotinate (1.1 g, 1.88 mmol) in DMF (11 mL) was added K2CO3 (0.39 g, 2.82 mmol) followed by the compound methanamine (1.1 mL, 2.26 mmol) and the reaction mixture was stirred at room temperature for 4 hours. After completion of the starting material (TLC monitoring), the reaction mixture was quenched with cold water and the precipitated solid was filtered off and dried under vacuum to give 0.82 g of compound methyl (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-(methylamino)nicotinate as an off-white solid. [TLC system: EtOAc:petroleum ether (1:9); R f Value: 0.3].

[0197] Step 2 (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-(methylamino)nicotinic acid (Int-15): [ka]

[0198] To a stirred solution of compound methyl (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-(methylamino)nicotinate (0.6 g, 1.01 mmol) in MeOH (3 mL), THF (3 mL) and water (1 mL) was added NaOH (0.081 g, 2.02 mmol) at 0° C. The reaction mixture was stirred at 50° C. for 3 hours. After completion of starting material (TLC monitoring), the reaction mixture was concentrated under reduced pressure to obtain crude compound. The crude compound was diluted with water and acidified with 1N HCl. The precipitated solid was filtered off and dried under vacuum to obtain 0 g of compound as an off-white solid. [TLC system: MeOH:DCM (1:9); f Value: 0.1].

[0199] Synthesis of 7-azidoheptan-1-amine (Int-16) [ka]

[0200] Step 1. 1,7-Diazidoheptane: [ka]

[0201] To a solution of compound 1,7-dibromoheptane (10 g, 38.76 mmol) in DMF (100 mL) was added NaN3 (5.04 g, 77.52 mmol) portionwise at room temperature. The reaction mixture was stirred at 60 °C for 16 h. After completion of the reaction (TLC monitoring). The reaction mixture was diluted with water (300 mL) and extracted with EtOAc (2 x 100 mL). The combined organic layers were dried over anhydrous sodium sulfate and evaporated to obtain the crude product. The obtained crude product was purified by silica gel (100-200 mesh) column chromatography using 3% EtOAc in petroleum ether to obtain 1,7-diazidoheptane (6.5 g) as a colorless oily compound. [TLC system: EtOAc: petroleum ether (1:9); R f Value: 0.8].

[0202] Step 2. 7-Azidoheptan-1-amine (Int-16): [ka]

[0203] To a solution of compound 1,7-diazidoheptane (6.5 g, 35.67 mmol) in 1N HCl (100 mL), EtOAc (32.5 mL) and Et2O (32.5 mL) were added followed by TPP (4.68 g, 17.84 mmol) and the reaction mixture was stirred at room temperature for 16 h. After completion of the reaction (TLC monitoring). The reaction mixture was diluted with water (200 mL) and washed with Et2O (200 mL). The aqueous layer was basified with saturated NaHCO3 and extracted with EtOAc (2 x 300 mL). The combined organic layers were dried over anhydrous sodium sulfate and evaporated to give 7-azidoheptane-1-amine (2.5 g) as a colorless oily compound. [TLC system: MeOH:DCM (1:9); R f Value: 0.1].

[0204] Synthesis of (1R,1'R,2S,2'S)-3,3'-(prop-2-yn-1-ylazanediyl)bis(1-((4R,5R)-5-hydroxy-2-phenyl-1,3-dioxan-4-yl)propane-1,2-diol) (Int-17): [ka]

[0205] To a solution of compound prop-2-yn-1-amine (0.2 g, 1.81 mmol) and (2R,3R)-2,3-dihydroxy-3-((4R,5R)-5-hydroxy-2-phenyl-1,3-dioxan-4-yl)propanal (0.97 g 3.62 mmol) in MeOH (20 mL), AcOH (0.1 mL) was added and the reaction mixture was stirred at 60° C. for 1 h. The reaction mixture was brought to room temperature and then NaCNBH3 (0.68 g 10.86 mmol) was added in portions at 0° C. The reaction mixture was stirred at 60° C. for 16 h. After completion of the reaction (TLC monitoring), the reaction mixture was concentrated under reduced pressure to obtain crude mass. The crude compound was washed with Et2O (50 mL) and dried under vacuum to give the compound (1R,1'R,2S,2'S)-3,3'-(prop-2-yn-1-ylazanediyl)bis(1-((4R,5R)-5-hydroxy-2-phenyl-1,3-dioxan-4-yl)propane-1,2-diol) (0.9 g) as an off-white solid. [TLC system: MeOH:DCM (1:9); R f Value: 0.1].

[0206] Synthesis of methyl 6-((7-(4-((bis((2S,3R)-2,3-dihydroxy-3-((4R,5R)-5-hydroxy-2-phenyl-1,3-dioxan-4-yl)propyl)amino)methyl)-1H-1,2,3-triazol-1-yl)heptyl)amino)-2-((7-(but-2-yn-1-yl)-8-((R)-3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)nicotinate (Int-18) [ka]

[0207] Step 1 Methyl (R)-6-((7-azidoheptyl)amino)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)nicotinate: [ka]

[0208] To a solution of compound methyl (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-fluoronicotinate (3.5 g, 6.00 mmol) in DMF (30 mL), compound 3 (1.41 g, 9.00 mmol) and K2CO3 (1.24 g, 9.00 mmol) were added and the reaction mixture was stirred at 80° C. for 16 hours. After completion of the reaction (TLC monitoring), the reaction mixture was diluted with water (100 mL) and extracted with EtOAc (2×100 mL). The combined organic layer was dried over anhydrous sodium sulfate and evaporated to obtain the crude product. The resulting crude product was purified by silica gel (100-200 mesh) column chromatography using 30% EtOAc in petroleum ether to give methyl (R)-6-((7-azidoheptyl)amino)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)nicotinate (2.8 g) as an off-white solid. [TLC system: EtOAc:petroleum ether (1:1); R f Value: 0.5].

[0209] Step 2. Methyl 6-((7-(4-((bis((2S,3R)-2,3-dihydroxy-3-((4R,5R)-5-hydroxy-2-phenyl-1,3-dioxan-4-yl)propyl)amino)methyl)-1H-1,2,3-triazol-1-yl)heptyl)amino)-2-((7-(but-2-yn-1-yl)-8-((R)-3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)nicotinate (Int-18): [ka]

[0210] The compounds methyl (R)-6-((7-azidoheptyl)amino)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)nicotinate (0.5 g, 0.69 mmol) and (1R,1'R,2S,2'S)-3,3'-(prop-2-yn-1-ylazanediyl)bis(1-((4R,5R)-5-hydroxy-2-phenyl-1,3-dioxan-4-yl)propane-1,2-diol) (0.97 g To a solution of CuSO4.5H2O (0.207 g, 0.83 mmol) and Na-ascorbate (0.204 g 1.03 mmol) in t-BuOH (10 mL) and H2O (1 mL), CuSO4.5H2O (0.207 g, 0.83 mmol) and Na-ascorbate (0.204 g 1.03 mmol) were added and the reaction mixture was stirred at room temperature for 16 h. After completion of the reaction (TLC monitoring). The reaction mixture was concentrated under reduced pressure to obtain a crude mass. The crude compound was washed with EtO (50 mL) and dried under vacuum to give compound methyl 6-((7-(4-((bis((2S,3R)-2,3-dihydroxy-3-((4R,5R)-5-hydroxy-2-phenyl-1,3-dioxan-4-yl)propyl)amino)methyl)-1H-1,2,3-triazol-1-yl)heptyl)amino)-2-((7-(but-2-yn-1-yl)-8-((R)-3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)nicotinate (0.9 g) as an off-white solid. The crude compound was taken to the next step without further purification. [TLC system: MeOH:DCM (1:9); R f Value: 0.1].

[0211] Synthesis of methyl 2-(bromomethyl)-5-chlorobenzoate (Int-19) [ka]

[0212] Methyl 5-chloro-2-methylbenzoate To a stirred solution of methyl 5-chloro-2-methylbenzoate (10.0 g, 54.16 mmol) in 1,2-dichloroethane (100 mL) was added N-bromosuccinimide (9.64 g, 54.16 mmol) and catalytic benzoyl peroxide (0.66 g, 2.71 mmol) in small portions. The reaction mixture was stirred at 80 °C for 5 h. After completion of the starting material, the reaction mixture was evaporated under reduced pressure, quenched with water and extracted with ethyl acetate (3 x 100 mL). The organic layer was washed with brine solution (150 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude compound. The crude compound was purified by column chromatography (silica gel 230-400 mesh, 100% petroleum ether used as eluent) to obtain compound methyl 2-(bromomethyl)-5-chlorobenzoate 8 g as a colorless liquid. [TLC system: petroleum ether; R f Value: 0.7].

[0213] Synthesis of (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoic acid (Int-20) [ka]

[0214] Step 1. 8-Bromo-7-(but-2-yn-1-yl)-3-methyl-3,7-dihydro-1H-purine-2,6-dione: [ka]

[0215] To a stirred solution of compound 8-bromo-3-methyl-3,7-dihydro-1H-purine-2,6-dione (10.0 g, 40.81 mmol) in DMF (100 mL), DIPEA (7.13 mL, 40.81 mmol) and 1-bromobut-2-yne (5.43 g, 40.81 mmol) were added portionwise at room temperature. The reaction mixture was stirred at room temperature for 16 hours. After completion of the starting material, the reaction mixture was quenched with cold water, and the precipitated solid was filtered off and dried under vacuum to give 9.3 g of compound 8-bromo-7-(but-2-yn-1-yl)-3-methyl-3,7-dihydro-1H-purine-2,6-dione as an off-white solid. [TLC system: MeOH:DCM (1:9); R f Value: 0.7].

[0216] Step 2. Methyl 2-((8-bromo-7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoate: [ka]

[0217] To a stirred solution of compound 8-bromo-7-(but-2-yn-1-yl)-3-methyl-3,7-dihydro-1H-purine-2,6-dione (2.0 g, 6.73 mmol) in DMF (20 mL), compound methyl 2-(bromomethyl)-5-chlorobenzoate (1.77 g, 6.73 mmol) and K2CO3 (2.79 g, 20.19 mmol) were added, and the reaction mixture was stirred at 50° C. for 16 hours. After completion of the starting material, the reaction mixture was quenched with cold water, and the precipitated solid was filtered off and dried under vacuum to give 2.5 g of compound methyl 2-((8-bromo-7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-5-chlorobenzoate as an off-white solid. [TLC system: EtOAc:petroleum ether (1:1); f Value: 0.7].

[0218] Step 3 Ethyl methyl (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoate: [ka]

[0219] To a stirred solution of compound methyl 2-((8-bromo-7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoate (2.5 g, 5.21 mmol) in DMF (25 mL) was added compound tert-butyl (R)-piperidin-3-ylcarbamate (1.25 g, 6.25 mmol) and K2CO3 (2.16 g, 15.63 mmol) and the reaction mixture was stirred at 65° C. for 8 h. After completion of the starting material, the reaction mixture was quenched with cold water, and the precipitated solid was filtered off and dried under vacuum to give 2.0 g of compound ethyl methyl (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoate as an off-white solid. [TLC system: EtOAc:petroleum ether (1:1); R f Value: 0.3].

[0220] Step 4 (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoic acid (Int-20): [ka]

[0221] To a stirred solution of compound ethyl methyl (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoate (0.5 g, 0.83 mmol) in MeOH (5 mL) and water (1 mL) was added LiOH.H2O (0.11 g, 2.5 mmol) at 0° C. The reaction mixture was stirred at room temperature for 16 hours. After completion of the starting material, the reaction mixture was concentrated under reduced pressure to obtain the crude compound. The crude compound was diluted with water and acidified with 1N HCl. The precipitated solid was filtered off and dried under vacuum to give 0.4 g of the compound (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoic acid as an off-white solid. [TLC system: MeOH:DCM (1:9); R f Value: 0.3].

[0222] Synthesis of (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylic acid (Int-21) [ka]

[0223] Step 1 Methyl (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate [ka]

[0224] To a stirred suspension of compound (R)-3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoic acid (1.35 g, 4.05 mmol) in DCM (20 mL) was added Et3N (1.69 mL, 12.15 mmol) and BOP-Cl (1.54 g, 6.07 mmol) followed by compound methyl 3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (1.0 g, 4.05 mmol). The reaction was stirred at room temperature for 16 h and the progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain the crude compound. The crude compound was purified by column chromatography on silica gel (Davisil) (using 0-60% EtOAc in petroleum ether as eluent) to obtain 1.6 g of compound methyl (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5 trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate as a white solid. [TLC system: EtOAc:petroleum ether (6:4); R f Value: 0.5].

[0225] Step 2 (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylic acid (Int-21): [ka]

[0226] To a solution of compound methyl (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5 trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (0.5 g, 0.88 mmol) in MeOH (10 mL) and THF (10 mL) was added 4 M NaOH solution (2.7 mL, 5.5 V) at 0° C. The reaction was then stirred at room temperature for 2 h. After completion of the reaction, the reaction mixture was concentrated and the residue was acidified with 10% HCl to pH-4, then the reaction mixture was concentrated to give a residue which was dried by co-evaporation with ACN and toluene to give 0.450 g of compound (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5 trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylic acid as an off-white solid.

[0227] Synthesis of 2-(2-(2-azidoethoxy)ethoxy)ethyl (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (Int-22) [ka]

[0228] To a stirred suspension of the compound (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylic acid (0.45 g, 0.81 mmol) in DCM (10 mL) was added EtN (0.341 mL, 2.45 mmol) and BOP-Cl (0.312 g, 1.22 mmol) followed by the compound 2-(2-(2-azidoethoxy)ethoxy)ethan-1-ol (0.214 g, 1.22 mmol). The reaction mixture was stirred at room temperature for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was concentrated to obtain the crude compound, which was purified by column chromatography on silica gel (Davisil) (using 0-60% EtOAc in petroleum ether as eluent) to obtain 0.4 g of compound 2-(2-(2-azidoethoxy)ethoxy)ethyl (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate as a colorless gum. [TLC system: EtOAc:petroleum ether (7:3); R f Value: 0.7].

[0229] Synthesis of 2-(2-(2-azidoethoxy)ethoxy)ethyl (R)-7-(3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (Int-23) [ka]

[0230] To a stirred solution of 2-(2-(2-azidoethoxy)ethoxy)ethyl (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (0.45 g, 0.64 mmol) in 1,4-dioxane (4.5 mL), 4M HCl in 1,4-dioxane (1 mL) was added dropwise at 0° C. and the reaction mixture was stirred at room temperature for 2 hours. After completion of the starting material, the reaction mixture was concentrated under reduced pressure to obtain the crude compound. The crude compound was purified by reverse phase preparative HPLC to obtain 0.1 g of compound Int-23 as an off-white solid [TLC system: EtOAc:petroleum ether (7:3); R f Value: 0.2].

[0231] Exemplary Compounds of the Disclosure [Example] [Example 1] Synthesis of heptyl (3-((2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl)carbamate (1) [ka]

[0232] Step 1. Heptyl (3-aminoadamantan-1-yl)carbamate: [ka]

[0233] Adamantane-1,3-diamine dihydrochloride [500 mg, 2.09 mmol], heptyl chloroformate [336 mg, 1.88 mmol] and TEA [316 mg, 3.14 mmol] were added in DCM [15 mL]. The reaction mixture was stirred at room temperature under N2 for 2 h. The mixture was filtered and washed three times with DCM (5 mL). The filtrate was concentrated to give methyl 6-chloro-4-methoxypyridazine-3-carboxylate (270 mg, 14% yield) as a white solid. ES MS M / Z=309(M+1) Step 2. Heptyl (3-((2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl)carbamate (1): [ka]

[0234] Heptyl N-(3-aminoadamantan-1-yl)carbamate [300 mg, 0.97 mmol], (2S)-1-(2-chloroacetyl)pyrrolidine-2-carbonitrile [150 mg, 0.87 mmol], K2CO3 [402 mg, 2.91 mmol] and potassium iodide [161 mg, 0.97 mmol] were added in MeCN [10 mL]. The reaction mixture was stirred at room temperature for 2 h. The mixture was concentrated and the residue was purified by column chromatography on silica gel eluted with (DCM:MeOH=20:1) to give heptyl (3-((2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl)carbamate (1,160 mg, 90% yield) as a yellow oil. ES MS M / Z=445(M+1) 1 H NMR(400 MHz,DMSO-d6)6.79(s,1H),4.73(dd,J=7.3,3.8 Hz,1H),3.85(t,J=6.4 Hz,2H),3.65-3.55(m,1H),3.51-3.38(m,2H),3.36(d,J=7.3 Hz,1H),3.29(s,1H),2.17-2.06(m,4H),2.05-1.95(m,2H),1.72(d,J=17.5 Hz,6H),1.49(d,J=12.8 Hz,8H),1.26(s,8H),0.86(t,J=6.9 Hz,3H). [Example 2] Synthesis of (2S)-1-((3-(heptylthio)adamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (2): [ka]

[0235] Step 1 Benzyl (3-hydroxyadamantan-1-yl)carbamate: [ka]

[0236] To a solution of 3-aminoadamantan-1-ol (5 g, 29.9 mmol) and DIEA (5.79 g, 44.9 mmol) in DCM (20 mL) was added a solution of CbzCl (5.08 g, 29.9 mmol) in DCM (20 mL) at 0 °C. The reaction mixture was stirred at 25 °C for 1 h. The mixture was diluted with water and extracted with DCM (100 mL). The organic layer was dried over Na2SO4 and concentrated under pressure at 40 °C. The residue was purified by flash column chromatography eluting with (MeOH / DCM, 5%-10%) to give benzyl (3-hydroxyadamantan-1-yl)carbamate (8.5 g, 94% yield) as a white solid. ES MS M / Z=302 (M+1) Step 2. 3-(((benzyloxy)carbonyl)amino)adamantan-1-yl methanesulfonate: [ka]

[0237] To a solution of benzyl (3-hydroxyadamantan-1-yl)carbamate (3.01 g, 10 mmol) and triethylamine (3 g, 30 mmol) in DCM (20 mL) was added a solution of methanesulfonyl chloride (1.71 g, 15 mmol) in DCM (2 mL) at 0° C. The reaction mixture was stirred at 0° C. for 1 h. The mixture was diluted with water and extracted with DCM (30 mL). The organic layer was dried over Na2SO4 and concentrated under pressure at 30° C. to give 3-(((benzyloxy)carbonyl)amino)adamantan-1-yl methanesulfonate (3.3 g, 87% yield) as a white solid, which was used in the next step without further purification. ES MS M / Z=402 (M+23) Step 3S-(3-aminoadamantan-1-yl)ethanethioate: [ka]

[0238] A solution of 3-(((benzyloxy)carbonyl)amino)adamantan-1-yl methanesulfonate (2.5 g, 6.59 mmol) in thiolacetic acid (15 mL) was stirred at 100° C. for 12 h. The reaction mixture was concentrated under pressure at 45° C. The residue was purified by flash column chromatography eluting with (MeOH / DCM, 5%-10%) to give S-(3-aminoadamantan-1-yl)ethanethioate (1 g, 67% yield) as a yellow solid. ES MS M / Z=226(M+1). Step 4S-(3-((2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl)ethanethioate: [ka]

[0239] To a mixture of S-(3-aminoadamantan-1-yl)ethanethioate (113 mg, 0.5 mmol), K2CO3 (207 mg, 1.5 mmol) and KI (8 mg, 0.05 mmol) in MeCN (5 mL) was added 2-chloro-1-(pyrrolidin-1-yl)ethan-1-one (58 mg, 0.4 mmol). The reaction mixture was stirred at 75 °C for 6 h. The reaction mixture was concentrated under pressure at 40 °C. The residue was purified by flash column chromatography eluting with MeOH / DCM (5%-10%) to give S-(3-((2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl)ethanethioate (128 mg, 71% yield) as a yellow oil. ES MS M / Z=362(M+1) Step 5 (2S)-1-((3-mercaptoadamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile: [ka]

[0240] To a solution of S-(3-((2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl)ethanethioate (1 g, 2.77 mmol) in MeOH / H2O (V / V=20:1, 10 mL) was added K2CO3 (1.14 g, 8.31 mmol). The reaction mixture was stirred at 25 °C for 30 min. The reaction mixture was dried over Na2SO4 and concentrated under pressure at 35 °C. The residue was purified by flash column chromatography eluting with (MeOH / DCM, 10%-20%) to give (2S)-1-((3-mercaptoadamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (300 mg, 33% yield) as a yellow oil. ES MS M / Z=320 (M+1) Step 6 tert-Butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)(3-mercaptoadamantan-1-yl)carbamate: [ka]

[0241] A mixture of (2S)-1-((3-mercaptoadamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (400 mg, 1.25 mmol), Boc2O (545 mg, 2.5 mmol) and TEA (631 mg, 6.25 mmol) in toluene (10 mL) was stirred at reflux for 12 h. The mixture was concentrated. The residue was purified by flash column chromatography eluting with MeOH / DCM (5%-10%) to give tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)(3-mercaptoadamantan-1-yl)carbamate (400 mg, 76% yield) as a colorless oil. ES MS M / Z=442 (M+23). Step 7 tert-Butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)(3-(heptylthio)adamantan-1-yl)carbamate: [ka]

[0242] A mixture of tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)(3-mercaptoadamantan-1-yl)carbamate (170 mg, 0.40 mmol), 1-iodoheptane (138 mg, 0.61 mmol) and potassium carbonate (224 mg, 1.62 mmol) in MeCN (3 mL) was stirred at 50° C. for 15 h. The mixture was concentrated and the residue was purified by silica gel column chromatography eluting with (PE:EtOAc=2:1) ​​to give tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)(3-(heptylthio)adamantan-1-yl)carbamate (50 mg, 27% yield) as a yellow solid. ES MS M / Z=540(M+23). Step 8 (2S)-1-((3-(heptylthio)adamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (2): [ka]

[0243] A solution of tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)(3-(heptylthio)adamantan-1-yl)carbamate (50 mg, 0.10 mmol) in TFA / DCM (V / V=1:3, 4 mL) was stirred at room temperature for 30 min. The mixture was concentrated and the residue was purified by preparative HPLC (Gemini-C18 150×21.2 mm, 5 um, mobile phase: ACN-H2O, gradient: 30-70%) to give (2S)-1-((3-(heptylthio)adamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (32 mg, 80% yield) as a white solid. ES MS M / Z=418(M+1) 1H NMR(400 MHz,CD3OD)8.40(s,1H),4.81(t,J=5.2 Hz,1H),3.99-3.87(m,2H),3.74-3.68(m,1H),3.56-3.49(m,1H),2.56(t,J=7.2 Hz,2H),2.35-2.24(m,4H),2.23-2.14(m,2H),2.01-1.94(m,2H),1.92-1.78(m,8H),1. 73-1.65(m,2H),1.57-1.50(m,2H),1.43-1.36(m,2H),1.35-1.24(m,6H),0.90(t,J=6.8 Hz,3H). [Example 3] Synthesis of (2S)-1-(((1S,3R,5S)-3-(heptylsulfinyl)adamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (3) [ka]

[0244] To a solution of tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(heptylthio)adamantan-1-yl)carbamate (20 mg, 0.03 mmol) in DCM (3 mL) under nitrogen atmosphere at -40° C. was added 3-chloroperbenzoic acid (10 mg, 0.05 mmol) dropwise. The reaction mixture was stirred at -40° C. for 30 min. TFA (1 mL) was added slowly. The reaction mixture was stirred at room temperature for an additional 30 min. The reaction mixture was concentrated under pressure. The crude product was then purified by Genal preparative HPLC (Gemini-C18 150×21.2 mm, 5 um, mobile phase: ACN-HO, 0.1% FA, gradient: 5-50%) to give the product (2S)-1-((3-(heptylsulfinyl)adamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (6.5 mg, 38.9%) as a white solid. MS(ESI):mass calcd.for C24H39N3O2S 433.66,m / z found 433.8 [M+H]+.1H NMR(400 MHz,CD3OD)8.38(s,1H),4.82-4.79(m,1H),3.95-3.84(m,2H),3.75-3.70(m,1H),3.56-3.50(m,1H),2.72-2.66(m,2H),2.43(s,2H), 2.31-2,25(m,2H),2.22-2.14(m,2H),2.01-1.72(m,14H),1.57-1.45(m,2H),1.42-1.35(m,2H),1.33-1.31(m,4H),0.92-0.89(m,3H). [Example 4] Synthesis of 2-(((1R,3S,5S)-3-((2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl)thio)ethyl(2-(4-(2-(hexyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethyl)phenoxy)ethyl)carbamate (4) [ka]

[0245] Step 1. tert-Butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)(3-((2-hydroxyethyl)thio)adamantan-1-yl)carbamate To a mixture of tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)(3-mercaptoadamantan-1-yl)carbamate (350 mg, 0.83 mmol) and K2CO3 (572 mg, 4.15 mmol) in MeCN (15 mL) was added 2-iodoethanol (1.42 g, 8.3 mmol). The reaction mixture was stirred at 25 °C for 12 h. The reaction mixture was concentrated under pressure. The residue was purified by flash chromatography eluting with CHCl2 / MeOH (10:1) to give the product tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)(3-((2-hydroxyethyl)thio)adamantan-1-yl)carbamate (340 mg, 88%) as a yellow oil. MS(ESI): mass calcd. for C 24 H 37 N3O4S 463.25,m / z found 486.2 [M+Na] + . Step 2. tert-Butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)(3-((2-(((4-nitrophenoxy)carbonyl)oxy)ethyl)thio)adamantan-1-yl)carbamate To a mixture of tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)(3-((2-hydroxyethyl)thio)adamantan-1-yl)carbamate (340 mg, 0.73 mmol) and EtN (221 mg, 2.19 mmol) in DCM (20 mL) was added 4-nitrophenyl chloroformate (220 mg, 1.09 mmol). The reaction mixture was stirred at 25° C. for 3 h. H2O (20 mL) was added. The residue was extracted with DCM (30 mL). The DCM phase was concentrated to give tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)(3-((2-(((4-nitrophenoxy)carbonyl)oxy)ethyl)thio)adamantan-1-yl)carbamate (600 mg, crude) as a yellow oil, which was used in the next step without further purification. MS(ESI): mass calcd. for C 31 H 40 N4O8S 628.26,m / z found 629.7 [M+H] + . Step 3. tert-Butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-((2-(((2-(4-(2-(hexyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethyl)phenoxy)ethyl)carbamoyl)oxy)ethyl)thio)adamantan-1-yl)carbamate To a solution of (2R,3R,4R,5S)-6-((4-(2-aminoethoxy)phenethyl)(hexyl)amino)hexane-1,2,3,4,5-pentaol (410 mg, 0.95 mmol) in THF (15 mL) was added TEA (303 mg, 3 mmol) followed by a solution of tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)(3-((2-(((4-nitrophenoxy)carbonyl)oxy)ethyl)thio)adamantan-1-yl)carbamate (903 mg, 1.43 mmol) in THF (5 mL). The reaction mixture was stirred at 25° C. for 4 h. The reaction mixture was concentrated under pressure. The residue was purified by flash chromatography, eluting with CHCl / MeOH (5:1) to give the product tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-((2-(((2-(4-(2-(hexyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethyl)phenoxy)ethyl)carbamoyl)oxy)ethyl)thio)adamantan-1-yl)carbamate (220 mg, 25%) as a yellow oil. MS(ESI): mass calcd. for C 47 H 75 N5O 11 S 917.52,m / z found 918.4 [M+H] + . Step 4. 2-(((1R,3S,5S)-3-((2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl)thio)ethyl(2-(4-(2-(hexyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethyl)phenoxy)ethyl)carbamate To a solution of tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-((2-(((2-(4-(2-(hexyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethyl)phenoxy)ethyl)carbamoyl)oxy)ethyl)thio)adamantan-1-yl)carbamate (400 mg, 0.43 mmol) in DCM (15 mL) was added ZnBr2 (1086 mg, 4.8 mmol). The reaction mixture was stirred at 25 °C for 8 h. The solution was filtered and the filtrate was concentrated and purified by preparative TLC (CHCl / MeOH)=(5:1) to give impure product, which was then purified by Genal preparative HPLC (Gemini-C18 150×21.2 mm, 5 um, mobile phase: ACN-HO, 0.1% FA, gradient: 5-50%) to give product 2-(((1R,3S,5S)-3-((2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl)thio)ethyl (2-(4-(2-(hexyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethyl)phenoxy)ethyl)carbamate (V2209613) (20 mg, 5%) as a white solid. MS(ESI):mass calcd.for C 42 H 67 N5O9S 817.47,m / z found 818.4 [M+H] + . 1 H NMR(400 MHz,MeOD)δ 8.53(s,2H),7.24(d,J=8.1 Hz,2H),6.94(d,J=8.2 Hz,2H),4.77-4.86(m,1H),4.24-3.97(m,5H),3.94-3.78(m,4H),3.77-3.63(m,4H),3.61-3.45(m,3H),3.44-3.33(m,5H),3.22-3.15(m,2 H),3.10-2.95(m,2H),2.88-2.73(m,2H),2.40-2.10(m,6H),2.02-1.92(m,2H),1.88-1.66(m,11H),1.45-1.30(m,6H),1.00-0.90(m,3H). [Example 5] Synthesis of (2S)-1-(((1S,3R,5S)-3-(2-hydroxyethoxy)adamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (5) [ka]

[0246] Step 1 tert-Butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-hydroxyethoxy)adamantan-1-yl)carbamate: [ka]

[0247] To a stirred solution of (1R,3S,5S)-3-((tert-butoxycarbonyl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl methanesulfonate (Int-2, 0.59 g, 1.22 mmol) and ethane-1,2-diol (3.8 g, 61.25 mmol) in acetonitrile (3.0 mL), molecular sieves 4 Å (2.0 g) were added and the resulting mixture was stirred at 70° C. for 16 h. After completion, the reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (3×50 mL). The combined organic layers were dried over anhydrous sodium sulfate and evaporated to give the crude product. The crude product was then purified by silica gel column chromatography using 3% methanol in dichloromethane and further purified by RP-HPLC using the following conditions: Column / dimensions: X-BRIDGE-C 18 (19*250, 5um), Mobile phase A: 10 mM ammonium bicarbonate in water, Mobile phase B: ACN Gradient (time / % of B): 00 / 10, 3 / 10, 7 / 35, 20 / 75, 20.1 / 100, 22 / 100, 22.1 / 10, 24 / 10, Flow rate: 17ml / min. Evaporation of the desired fractions gave tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-hydroxyethoxy)adamantan-1-yl)carbamate (0.1g) as an off-white solid. TLC system: MeOH:DCM (1:9); f :0.4 Step 2 (2S)-1-(((1S,3R,5S)-3-(2-hydroxyethoxy)adamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (5): [ka]

[0248] To a stirred suspension of tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-hydroxyethoxy)adamantan-1-yl)carbamate (0.09 g, 0.2 mmol) in acetonitrile (5 mL) was added 1 M tin(IV) chloride solution in heptane (0.8 mL, 0.8 mmol) and the mixture was stirred at room temperature for 3 h. After completion, the reaction mixture was evaporated and the residue was co-evaporated with methanol (10 mL) and washed with diethyl ether (2×5 mL) to give the crude product. The crude product was then purified by C-18 column chromatography using 30% methanol in 10 mM ammonium bicarbonate in water. The desired fractions were lyophilized to give (2S)-1-(((1S,3R,5S)-3-(2-hydroxyethoxy)adamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (0.055 g) as an off-white solid. TLC system: MeOH:DCM (0.5:9.5); f :0.1LCMS M / Z 348.37(M+1) 1 H NMR(400 MHz,DMSO-d6)δ 4.74-4.72(m,1H),4.46(bs,1H),3.63-3.32(m,8H),2.17-2.11(m,4H),2.03-1.99(m,2H),1.68(t,J=6.0 Hz,1H),1.60-1.40(m,12H). [Example 7] Synthesis of ((3-((2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl)oxy)methyl octanoate (7): [ka]

[0249] Step 1. Benzyl (3-((methylthio)methoxy)adamantan-1-yl)carbamate: [ka]

[0250] A mixture of benzyl (3-hydroxyadamantan-1-yl)carbamate (3 g, 9.97 mmol) and Ac2O (35.1 g, 29.9 mmol) in DMSO (30 mL) was stirred at room temperature overnight. The reaction mixture was diluted with EtOAc (100 ml) and washed with water (30 mL x 3). The organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The crude product was purified by column chromatography on silica gel eluted with (PE / EtOAc = 8 / 1) to give benzyl (3-((methylthio)methoxy)adamantan-1-yl)carbamate (2.6 g, 72.2% yield) as a pale yellow oil. ES MS M / Z = 384 (M+23) Step 2. ((3-(((benzyloxy)carbonyl)amino)adamantan-1-yl)oxy)methyl octanoate: [ka]

[0251] To a solution of benzyl (3-((methylthio)methoxy)adamantan-1-yl)carbamate (2.5 g, 6.93 mmol) in CDCl3 (20 mL) was added SOCl2 (1.99 g, 13.86 mmol) slowly at 0 °C. The mixture was stirred at 0 °C for 30 min. The mixture was concentrated and dissolved in THF (20 mL). To this solution was added TEA (1.40 g, 13.86 mmol) and octanoic acid (1.99 g, 13.86 mmol). The reaction mixture was stirred at room temperature overnight. The mixture was diluted with EtOAc (50 ml) and washed with water (20 mL x 2). The organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with (PE / EtOAc=8 / 1) to give ((3-(((benzyloxy)carbonyl)amino)adamantan-1-yl)oxy)methyl octanoate (1.9 g, 60% yield) as a pale yellow oil. ES MS M / Z=480 (M+23). Step 3. ((3-aminoadamantan-1-yl)oxy)methyl octanoate: [ka]

[0252] To a solution of -(((benzyloxy)carbonyl)amino)adamantan-1-yl)oxy)methyl octanoate (1.7 g, 3.72 mmol) in MeOH (10 mL) was added Pd / C (500 mg). The reaction mixture was stirred overnight at room temperature under H2 atmosphere. The mixture was filtered and the filtrate was evaporated under reduced pressure to give ((3-aminoadamantan-1-yl)oxy)methyl octanoate (1.1 g, 91.4% yield) as a colorless oil. ES MS M / Z=324(M+1) Step 4. ((3-((2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl)oxy)methyl octanoate (7): [ka]

[0253] A mixture of ((3-aminoadamantan-1-yl)oxy)methyl octanoate (1 g, 3.1 mmol), (S)-1-(2-chloroacetyl)pyrrolidine-2-carbonitrile (533 mg, 3.1 mmol), potassium carbonate (513 mg, 3.72 mmol) and potassium iodide (257 mg, 1.55 mmol) in acetonitrile (15 mL) was stirred at 50° C. for 5 h under N2 atmosphere. The mixture was diluted with EtOAc (30 ml) and washed with water (10 mL x 2). The organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with (PE / EtOAc=6 / 1) to give ((3-((2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl)oxy)methyl octanoate (mCMP694) (400 mg, 28% yield) as a pale yellow oil. ES MS M / Z=460(M+1). 1H NMR(400 MHz, CDCl3)5.39(s,2H),4.87-4.73(m,1H),3.66-3.39(m,4H),2.36-2.25(m,6H),2.24-2.13 (m,2H),1.83(s,3H),1.80-1.68(m,6H),1.66-1.56(m,6H),1.33-1.24(m,8H),0.88(t,J=7.2 Hz,3H). [Example 8] Synthesis of (2S)-1-(((1S,3R,5S)-3-(2-(2-(2-(4-((hexyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethoxy)adamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (8): [ka]

[0254] Step 1 tert-Butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)adamantan-1-yl)carbamate: [ka]

[0255] To a solution of the compound (1R,3S,5S)-3-((tert-butoxycarbonyl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl methanesulfonate (137 g, 285 mmol, 1.00 equiv.) in THF (1.40 L) was added TEA (66.1 g, 653 mmol, 2.29 equiv.) and 2,2'-(ethane-1,2-diylbis(oxy))bis(ethan-1-ol) (722 g, 4.81 mol, 16.8 equiv.). The reaction was heated to 70 °C for 48 h. The reaction mixture was diluted with HO (2.00 L) and extracted with EA (2.00 L x 2). The combined organic layers were washed with 5% aqueous citric acid (2.00 L), then the combined organic layers were washed with saturated aqueous NaHCO3 (2.00 L), dried over Na2SO4, filtered, and concentrated under reduced pressure below 45° C. The residue was purified by column chromatography (SiO2, PE / EA=1 / 1 to 0 / 1) to give the compound tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)adamantan-1-yl)carbamate (175 g, 327 mmol, yield 57.4%, purity N / A) as a pale yellow oil. TLC system: EA only; R f :0.15.

[0256] Step 2. 2-(2-(2-(((1R,3S,5S)-3-((tert-butoxycarbonyl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl)oxo)ethoxy)ethoxy)ethyl methanesulfonate: [ka]

[0257] To a solution of compounds tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)adamantan-1-yl)carbamate (162 g, 302 mmol, 1.00 equiv.) and N,N,N',N'-tetramethyl-1,6-hexanediamine (115 g, 667 mmol, 2.21 equiv.) in DCM (1600 mL) was added MsCl (54.9 g, 479 mmol, 37.1 mL, 1.59 equiv.) at 0-10 °C. The reaction was stirred at 0-10 °C for 2 h. The reaction mixture was quenched by the addition of ice water (1.00 L) at 0° C., then titrated with 5% aqueous citric acid (2.00 L) until pH=5-6, and extracted with EA (3.00 L). The combined organic layers were washed with NaHCO3 (1.00 L), dried over Na2SO4, filtered, and concentrated under reduced pressure to give compound 2-(2-(2-(((1R,3S,5S)-3-((tert-butoxycarbonyl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl)oxy)ethoxy)ethoxy)ethyl methanesulfonate (180 g, crude) as a pale yellow oil. TLC system: EA; R f :0.30.

[0258] Step 3. tert-Butyl ((1S,3R,5S)-3-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamate: [ka]

[0259] To a solution of compound 2-(2-(2-(((1R,3S,5S)-3-((tert-butoxycarbonyl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl)oxy)ethoxy)ethyl methanesulfonate (90.0 g, 146 mmol, 1.00 equiv.) in DMF (1.00 L) was added NaN3 (22.3 g, 343 mmol, 2.34 equiv.) under N2 at 20-30 °C, and the mixture was then stirred at 70 °C for 12 h. The reaction was added to saturated Na2CO3 solution (2.00 L) and then EA Extracted with 2.00 L of ethyl acetate. The combined organic layers were washed with 2.00 L of brine, dried over Na2SO4, filtered and concentrated under reduced pressure to give compound tert-butyl ((1S,3R,5S)-3-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamate (153 g, crude) as a pale yellow oil. TLC system: EA;R f :0.30.

[0260] Step 4 tert-Butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-(2-(2-(4-((hexyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethoxy)adamantan-1-yl)carbamate: [ka]

[0261] A mixture of compounds tert-butyl ((1S,3R,5S)-3-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamate (152 g, 271 mmol, 1.00 equiv.), (2R,3R,4R,5S)-6-(hexyl(prop-2-yn-1-yl)amino)hexane-1,2,3,4,5-pentaol (98.8 g, 325 mmol, 1.20 equiv.) and CuI (6.84 g, 35.9 mmol, 0.13 equiv.) in THF (1.52 L) was stirred at 25° C. for 12 h. The reaction mixture was diluted with HO (2.00 L) and extracted with EA (2.00 L). The combined organic layers were washed with brine (1.00 L), dried over Na2SO4, filtered and concentrated under reduced pressure. The crude product was purified by reverse phase HPLC (0.1% NH3·H2O) to give the compound tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-(2-(2-(4-((hexyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethoxy)adamantan-1-yl)carbamate (100 g, 115 mmol, 42.7% yield, 41.3% purity) as a yellow oil. TLC system: DCM:MeOH=5:1; R f :0.20. Step 5 (2S)-1-(((1S,3R,5S)-3-(2-(2-(2-(4-((hexyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethoxy)adamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (8): [ka]

[0262] To a solution of compound tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-(2-(2-(4-((hexyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethoxy)adamantan-1-yl)carbamate (40.0 g, 46.3 mmol, 1.00 equiv.) in ACN (320 mL) was added HCl (1 M, 560 mL, 12.1 equiv.) at 0° C. The reaction mixture was stirred at 25° C. for 18 h. The reaction mixture was extracted with DCM (1.00 L×2). The aqueous phase was adjusted to pH 9-10 with saturated NaHCO3 and K2CO3 and extracted with DCM (1.00 L x 2). The combined organic phase was dried over Na2SO4, filtered and concentrated under reduced pressure to give 13 (24.5 g, 31.0 mmol, 66.9% yield, 96.7% purity) as a pale yellow oil. LCMS M / Z M+1=764.7. 1 H-NMR(400 MHz,CDCl3)δ 7.73(s,1H),4.92-4.70(m,1H),4.52(br t,J=4.5 Hz,2H),3.94-3.82(m,5H),3.81-3.71(m,4H),3.71-3.63(m,3H),3.56(br d,J=19.3 Hz,9H),3.47-3.33(m,3H),2.77-2.58(m,2H),2.53(td,J=7.7,12.7 Hz,1H),2.47-2.37(m,1H),2.36-2.01(m,6H),1.99(s,1H),1.75-1.40(m,14H),1.23(br s,6H),0.84(br t,J=6.7 Hz,3H). Similarly, compound 15 was prepared from Int-1 and Int-5. Compounds 20 and 21 were prepared from saxagliptin following the route of compound 8.

[0263] [Example 9] Synthesis of (2S)-1-(((1S,3R,5S)-3-(2-(2-(4-((bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)adamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (9): [ka]

[0264] Step 1 tert-Butyl ((1S,3R,5S)-3-(2-(2-(4-((bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamate: [ka]

[0265] A mixture of tert-butyl ((1S,3R,5S)-3-(2-(2-azidoethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamate (0.6 g, 1.16 mmol), (2R,2'R,3R,3'R,4R,4'R,5S,5'S)-6,6'-(prop-2-yn-1-ylazanediyl)bis(hexane-1,2,3,4,5-pentaol) (0.66 g, 1.74 mmol), copper sulfate pentahydrate (0.35 g, 1.39 mmol) and sodium ascorbate (0.34 g, 1.74 mmol) in a mixture of tert-butyl alcohol (10 mL) and water (2 mL) was stirred at room temperature for 16 hours. After completion (monitored by LCMS), the reaction mixture was filtered through Celite, washed with methanol and water (1:1) (100 mL) and the filtrate was evaporated to give the crude product. The crude product was then purified by reverse phase C18 column chromatography using 50% methanol in 0.1% formic acid in water. Fractions containing the desired product were collected and evaporated to give tert-butyl ((1S,3R,5S)-3-(2-(2-(4-((bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamate (0.46 g, 44%) as an off-white solid.

[0266] Step 2 (2S)-1-(((1S,3R,5S)-3-(2-(2-(4-((bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)adamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (9): [ka]

[0267] To a stirred suspension of tert-butyl ((1S,3R,5S)-3-(2-(2-(4-((bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamate (0.35 g, 0.39 mmol) in acetonitrile (15 mL) was added a 1 M solution of tin(IV) chloride in heptane (1.55 mL, 1.55 mmol) and the reaction mixture was stirred at room temperature for 3 h. After completion (reaction monitored by LCMS), the reaction mixture was evaporated and the residue was co-evaporated with methanol (10 mL) and washed with diethyl ether (2×5 mL) to give the crude product. The crude product was then purified by C-18 column chromatography using 30% methanol in 10 mM ammonium bicarbonate in water. The desired fractions were lyophilized to give (2S)-1-(((1S,3R,5S)-3-(2-(2-(4-((bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)adamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (0.076 g) as a pale yellow solid. LCMS M / Z 800.59 (M+1). 1 H NMR(400 MHz,DMSO-d6)δ 7.95(s,1H),4.75-4.72(m,1H),4.70-4.10(m,12H),3.81-3.72(m,6H),3.63-3.39(m,22H),2.17-1.91(m,6H),1.56-1.44(m,12H). [Example 10] Synthesis of (2S)-1-(((1S,3R,5S)-3-(2-(2-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethoxy)ethoxy)adamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (10): [ka]

[0268] Step 1 tert-Butyl ((1S,3R,5S)-3-(2-(2-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamate: [ka]

[0269] To a mixture of tert-butyl ((1S,3R,5S)-3-(2-(2-aminoethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamate (0.21 g, 0.43 mmol), (2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanal (0.19 g, 1.07 mmol) and acetic acid (1.0 mL) in methanol (10 mL) was added sodium cyanoborohydride (0.034 g, 0.52 mmol). The reaction mixture was stirred at 70° C. for 24 hours. After completion of the reaction (reaction monitored by LCMS), the reaction mixture was evaporated to obtain the crude product. The obtained crude product was purified by reverse phase C18 column chromatography using 50% methanol in 0.1% formic acid in water. Fractions containing the desired product were collected and evaporated to give tert-butyl ((1S,3R,5S)-3-(2-(2-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamate (0.28 g) as a colourless gum.

[0270] Step 2 (2S)-1-(((1S,3R,5S)-3-(2-(2-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethoxy)ethoxy)adamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (10): [ka]

[0271] To a stirred solution of tert-butyl ((1S,3R,5S)-3-(2-(2-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamate (0.22 g, 0.27 mmol) in water (5 mL) was added 2N aqueous hydrochloric acid and the reaction mixture was stirred at room temperature for 24 hours. After completion of the reaction (monitored by LCMS), the reaction mixture was purified by RP Prep HPLC using the following conditions: Column / dimensions: X-BRIDGE(19*150), 5um, Mobile phase A: 10mM Ammonium Bicarbonate in water (aqueous), Mobile phase B: ACN, Gradient (time / % of B): 0 / 2, 3 / 2, 16 / 36, 16.1 / 100, 18 / 100, 18.1 / 2, 20 / 2, Flow rate: 17ml / min. The desired fractions were lyophilized to give (2S)-1-(((1S,3R,5S)-3-(2-(2-(bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethoxy)ethoxy)adamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (0.14 g, 72.5%) as an off-white solid. LCMS: M / Z 717.52 [M-1] 1 H NMR(400 MHz,DMSO-d6):δ 4.74-4.70(m,1H),4.51-4.46(m,4H),4.35-4.29(m,4H),4.19(d,J=6.4 Hz,2H),3.65-3.55(m,7H),3.49-3.33(m,16H),2.75-2.50(m,5H),2.17-1.95(m,6H),1.70(bs,1H),1.59-1.45(m,12H). [Example 11] Synthesis of (2S)-1-(((1S,3R,5S)-3-(2-(2-(((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethoxy)ethoxy)adamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (11): [ka]

[0272] Step 1 tert-Butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-(2-(((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethoxy)ethoxy)adamantan-1-yl)carbamate: [ka]

[0273] To a mixture of tert-butyl ((1S,3R,5S)-3-(2-(2-aminoethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamate (0.22 g, 0.45 mmol), (2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanal (0.081 g, 0.45 mmol) and acetic acid (1.0 mL) in methanol (50 mL) was added sodium cyanoborohydride (0.03 g, 0.45 mmol). The reaction mixture was stirred at 60° C. for 24 h. After completion (reaction monitored by LCMS), the mixture was evaporated under reduced pressure to give the crude product. The crude product was then purified by reverse phase preparative HPLC using the following conditions: Column / dimensions: X-BRIDGE(19*150), 5um, Mobile phase A: 10mM Ammonium Bicarbonate in water (aqueous), Mobile phase B: ACN, Gradient (time / % of B): 0 / 10, 1 / 10, 20 / 50, 20.10 / 95, 22.10 / 95, 22.20 / 10, 24 / 10 Flow rate: 17ml / min. The desired fractions were collected and lyophilized to give tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-(2-(((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethoxy)ethoxy)adamantan-1-yl)carbamate (0.035 g) as an off-white solid.

[0274] Step 2 (2S)-1-(((1S,3R,5S)-3-(2-(2-(((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethoxy)ethoxy)adamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (11): [ka]

[0275] To a stirred suspension of tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-(2-(((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethoxy)ethoxy)adamantan-1-yl)carbamate (0.03 g, 0.046 mmol) in dichloromethane (1 mL) was added trifluoroacetic acid at 0° C. and the reaction mixture was stirred at room temperature for 1 h. After completion (monitored by LCMS), the reaction mixture was evaporated under reduced pressure to give the crude product. The crude product was then purified by reverse phase preparative HPLC using the following conditions: Column / dimensions: X-BRIDGE(19*150), 5um, Mobile phase A: 10mM Ammonium Bicarbonate in water (aqueous), Mobile phase B: ACN, Gradient (time / % of B): 0 / 10, 1 / 10, 11 / 45.20, 11.10 / 95, 13.10 / 95, 13.20 / 10, 15 / 10, Flow rate: 17ml / min. The desired fractions were lyophilized to give (2S)-1-(((1S,3R,5S)-3-(2-(2-(((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethoxy)ethoxy)adamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (0.012 g, 47.2%) as an off-white solid. LCMS: M / Z 555.29 (M+1). 1 H NMR(400 MHz,DMSO-d6)δ 4.73-4.29(m,6H),3.63-3.13(m,17H),2.67-2.60(m,4H),2.16-1.99(m,6H),1.59-1.45(m,12H). [Example 12] Synthesis of 2-(2-(((1R,3S,5S)-3-((2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl)thio)ethyl(4-((ethyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)benzyl)carbamate (12) [ka]

[0276] Step 1 (2R,3R,4R,5S)-6-(ethylamino)hexane-1,2,3,4,5-pentanol: [ka]

[0277] To a mixture of (2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanal (5 g, 2.77 mmol) and 2M ethylamine in THF (20 mL, 41.6 mmol) in methanol (150 mL) was added Raney Nickel (5 g) and the reaction mixture was stirred at 70° C. under 150 psi of hydrogen gas for 16 h. After completion (monitored by LCMS), the reaction mixture was filtered through a Celite pad and washed with methanol (50 mL). The filtrate was evaporated to give a solid which was triturated with diethyl ether (50 mL) and dried to give (2R,3R,4R,5S)-6-(ethylamino)hexane-1,2,3,4,5-pentaol (5 g, 86%) as an off-white solid.

[0278] Step 2 tert-Butyl (4-((ethyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)benzyl)carbamate: [ka]

[0279] To a solution of (2R,3R,4R,5S)-6-(ethylamino)hexane-1,2,3,4,5-pentanol (1.0 g, 4.78 mmol) in methanol (10 mL) was added potassium carbonate (0.99 g, 7.17 mmol) followed by tert-butyl (4-(bromomethyl)benzyl)carbamate (1.44 g, 4.78 mmol) in portions at 0° C. and the mixture was stirred at room temperature for 16 h. After completion (monitored by LCMS), the reaction mixture was evaporated to give the crude product. The crude product was then purified by reversed-phase C18 flash column chromatography using 18% 0.1% formic acid in water and methanol as the mobile phase. Fractions containing the desired product were collected and evaporated to give tert-butyl (4-((ethyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)benzyl)carbamate formate (1.3 g) as a colorless foamy solid.

[0280] Step 3 (2R,3R,4R,5S)-6-((4-(aminomethoxy)benzyl)(ethyl)amino)hexane-1,2,3,4,5-pentanol hydrochloride: [ka]

[0281] To a stirred solution of tert-butyl (4-((ethyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)benzyl)carbamate formate (0.25 g, 0.527 mmol) in methanol (2.5 mL) was added 4 M hydrochloric acid in 1,4-dioxane slowly at 0° C. and the resulting mixture was stirred at room temperature for 2 h. After completion (monitored by LCMS), the reaction mixture was evaporated to give (2R,3R,4R,5S)-6-((4-(aminomethyl)benzyl)(ethyl)amino)hexane-1,2,3,4,5-pentaol hydrochloride (0.19 g, crude) as a green semi-solid.

[0282] Step 4. 2-(2-(((1R,3S,5S)-3-((2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl)oxo)ethoxy)ethyl (4-nitrophenyl)carbonate: [ka]

[0283] To a stirred solution of (2S)-1-(((1S,3R,5S)-3-(2-(2-hydroxyethoxy)ethoxy)adamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (0.2 g, 0.407 mmol) in dichloromethane (4 mL) was added triethylamine (0.164 g, 1.628 mmol) followed by 4-nitrophenyl carbonochloridate (0.09 g, 0.447 mmol) in dichloromethane (1 mL) at -10°C and the mixture was stirred at room temperature for 2 h. After completion, the reaction mixture was taken directly into the next step.

[0284] Step 5. 2-(2-(((1R,3S,5S)-3-((2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl)thio)ethyl(4-((ethyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)benzyl)carbamate: [ka]

[0285] To a stirred suspension of (2R,3R,4R,5S)-6-((4-(aminomethyl)benzyl)(ethyl)amino)hexane-1,2,3,4,5-pentaol hydrochloride (0.17 g, 0.47 mmol) in acetonitrile (5 mL) was added triethylamine (0.24 g, 2.34 mmol) followed by 2-(2-(((1R,3S,5S)-3-((2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl)oxy)ethoxy)ethyl (4-nitrophenyl)carbonate (0.26 g, 0.39 mmol) in dichloromethane (5 mL) at 0° C. and the mixture was stirred at room temperature for 16 h. After completion (monitored by LCMS), the reaction mixture was evaporated to give the crude product, which was then purified by reverse-phase C18 column chromatography using 40% 0.1% formic acid in water and acetonitrile as the mobile phase. The fractions containing the desired product were collected and evaporated to give 2-(2-(((1R,3S,5S)-3-((2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl)oxy)ethoxy)ethyl(4-((ethyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)benzyl)carbamate (0.3 g, crude) as a yellow semi-solid.

[0286] Step 6. 2-(2-(((1R,3S,5S)-3-((2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl)thio)ethyl(4-((ethyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)benzyl)carbamate(12-formate): [ka]

[0287] To a stirred suspension of 2-(2-(((1R,3S,5S)-3-((2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl)oxy)ethoxy)ethyl(4-((ethyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)benzyl)carbamate (0.29 g, 0.34 mmol) in dichloromethane (4 mL) was added trifluoroacetic acid at 0° C. and the reaction mixture was stirred at room temperature for 3 h. After completion (monitored by LCMS), the reaction mixture was concentrated and the residue was washed with diethyl ether (3×10 mL) and dried to give the crude product. The crude product was then purified by reverse phase preparative HPLC using the following conditions: Column / dimensions: SUNFIREC18 (19*150*5μ), Mobile phase A: 0.1% FA in water (aqueous), Mobile phase B: Acetonitrile, Gradient (time / % of B): 0 / 5, 2 / 5, 10 / 20, 10.1 / 100, 14 / 100, 14.1 / 5, 16 / 5, Flow rate: 17 ml / min. The desired fractions were lyophilized to give 2-(2-(((1R,3S,5S)-3-((2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl)oxo)ethoxy)ethyl(4-((ethyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)benzyl)carbamate formate (0.038 g) as an off-white solid. LCMS M / Z 746.72 (M+1). 1 H NMR(400 MHz,DMSO-d6):δ 8.17(s,2H),7.73(s,1H),7.27(d,J=7.6 Hz,2H),7.17(d,J=8.0 Hz,2H),4.75-4.74(m,3H),4.16-4.06(m,4H),3.75-3.35(m,21H),2.61-2.50( m,1H),2.46-2.41(m,3H),2.19-1.99(m,6H),1.60-1.52(m,12H),0.94(t,J=7.2 Hz,3H). [Example 13] Synthesis of 2-(((1R,3S,5S)-3-((2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl)oxy)ethyl(2-(4-(2-(hexyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethyl)phenoxy)ethyl)carbamate (13) [ka]

[0288] Step 1 tert-Butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-hydroxyethoxy)adamantan-1-yl)carbamate: [ka]

[0289] To a solution of (1R,3S,5S)-3-((tert-butoxycarbonyl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl methanesulfonate (1.25 g, 2.59 mmol) in MeCN (5 mL) was added ethane-1,2-diol (8.04 g, 129.6 mmol). The reaction mixture was stirred at 70° C. for 12 h. The reaction mixture was concentrated under pressure at 40° C. The residue was purified by flash column chromatography, eluting with (MeOH / DCM, 1%-10%) to give tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-hydroxyethoxy)adamantan-1-yl)carbamate (860 mg, 74% yield) as a yellow oil. ES MS M / Z=470 (M+23). Step 2 tert-Butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-(((4-nitrophenoxy)carbonyl)oxy)ethoxy)adamantan-1-yl)carbamate: [ka]

[0290] To a solution of tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-hydroxyethoxy)adamantan-1-yl)carbamate (860 mg, 1.92 mmol) and TEA (1000 mg, 10 mmol) in DCM (20 mL) was added 4-nitrophenyl chloroformate (800 mg, 4 mmol). The reaction mixture was stirred at 25 °C for 3 h. The mixture was quenched with H2O (20 mL) and extracted with DCM (30 mL). The organic layer was dried over Na2SO4 and concentrated under pressure for 30 h. o Concentration at 37° C. afforded tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-(((4-nitrophenoxy)carbonyl)oxy)ethoxy)adamantan-1-yl)carbamate (1.2 g, crude) as a yellow oil which was used in the next step without further purification. ES MS M / Z=635 (M+23). Step 2 tert-Butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-(((2-(4-(2-(hexyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethyl)phenoxy)ethyl)carbamoyl)oxy)ethoxy)adamantan-1-yl)carbamate [ka]

[0291] To a solution of (2R,3R,4R,5S)-6-((4-(2-aminoethoxy)phenethyl)(hexyl)amino)hexane-1,2,3,4,5-pentaol in THF (10 mL) was added TEA (210 mg, 2.1 mmol) followed by a solution of tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-(((4-nitrophenoxy)carbonyl)oxy)ethoxy)adamantan-1-yl)carbamate (428 mg, 0.7 mmol) in THF (1 mL). The reaction mixture was stirred at 25° C. for 4 h. The reaction mixture was concentrated under pressure at 40° C. The residue was purified by flash chromatography, eluting with (MeOH / DCM, 10%-50%) to give tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-(((2-(4-(2-(hexyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethyl)phenoxy)ethyl)carbamoyl)oxy)ethoxy)adamantan-1-yl)carbamate (436 mg, 69% yield) as a yellow oil. ES MS M / Z=902(M+1). Step 3. 2-(((1R,3S,5S)-3-((2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl)oxy)ethyl(2-(4-(2-(hexyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethyl)phenoxy)ethyl)carbamate (13): [ka]

[0292] To a solution of tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-(((2-(4-(2-(hexyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethyl)phenoxy)ethyl)carbamoyl)oxy)ethoxy)adamantan-1-yl)carbamate (436 mg, 0.48 mmol) in DCM (15 mL) was added ZnBr2 (1086 mg, 4.8 mmol). The reaction mixture was stirred at 25 °C for 12 h. The mixture was filtered and the filtrate was concentrated under pressure at 35 °C. The residue was purified by preparative TLC (CH2Cl2 / MeOH) = (5:1) to give impure product, which was purified by Genal preparative HPLC (Gemini-C18 150 x 21.2 mm, 5 um, mobile phase: ACN-H2O, 0.1% FA, gradient: 5-50%) to give 2-(((1R,3S,5S)-3-((2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)amino)adamantan-1-yl)oxy)ethyl(2-(4-(2-(hexyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethyl)phenoxy)ethyl)carbamate (13) (50 mg, 12%) as a white solid. ES MS M / Z = 802 (M+1). 1 H NMR(400 MHz,CD3OD)8.48(s,2H),7.22(d,J=8.5 Hz,2H),6.92(d,J=8.6 Hz,2H),4.83-4.78(m,1H),4.17-4.07(m,3H),4.05-3.95(m,2H),3.91- 3.74(m,4H),3.74-3.60(m,6H),3.55-3.50(m,1H),3.50-3.43(m,2H),3. 41-3.32(m,4H),3.26-3.16(m,2H),3.05-2.94(m,2H),2.40-2.09(m,6H) ,1.85-1.65(m,12H),1.58(s,2H),1.42-1.30(m,6H),0.97-0.89(m,3H). [Example 14] Synthesis of (2S)-1-(((1S,3R,5S)-3-(2-(2-(4-((((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)adamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (14) [ka]

[0293] Step 1 Mixture of tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-(2-(4-((((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)adamantan-1-yl)carbamate and tert-butyl ((1S,3R,5S)-3-(2-(2-(4-(aminomethyl)-1H-1,2,3 triazol-1-yl)ethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamate: [ka]

[0294] A mixture of tert-butyl ((1S,3R,5S)-3-(2-(2-azidoethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamate (1.0 g, 1.94 mmol), (2R,3R,4R,5S)-6-(prop-2-yn-1-ylamino)hexane-1,2,3,4,5-pentaol) (0.64 g, 2.90 mmol), copper(II) sulfate pentahydrate (0.58 g, 2.33 mmol) and sodium ascorbate (0.58 g, 2.91 mmol) in a mixture of tert-butyl alcohol (10 mL) and water (2 mL) was stirred at room temperature for 16 hours. After completion, the reaction mixture was filtered through Celite, washed with methanol and water (1:1) (100 mL) and the filtrate was evaporated to give the crude product. The crude product was then purified by reverse phase C18 column chromatography using 50% methanol in 0.1% formic acid in water. Fractions containing the desired product were collected and evaporated to give tert-butyl(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-(2-(4-((((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy. )adamantan-1-yl)carbamate and tert-butyl ((1S,3R,5S)-3-(2-(2-(4-(aminomethyl)-1H-1,2,3 triazol-1-yl)ethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamate (0.59 g, crude) was obtained as a brown solid.

[0295] Step 2 tert-Butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-(2-(4-((((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)adamantan-1-yl)carbamate: [ka]

[0296] To a mixture of tert-butyl ((1S,3R,5S)-3-(2-(2-(4-(aminomethyl)-1H-1,2,3 triazol-1-yl)ethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamate (0.44 g, 0.77 mmol), (2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanal (0.11 g, 0.61 mmol) and acetic acid (0.5 mL) in methanol (25 mL) was added sodium cyanoborohydride (0.048 g, 0.77 mmol) and the mixture was stirred at 60° C. for 6 h. After completion, the reaction mixture was evaporated to give the crude product which was then purified by RP preparative HPLC using the following conditions: Column / dimensions: X-BRIDGEC18(19*250) 5um, Mobile phase A: 10mM ABC in water (aqueous), Mobile phase B: 100% ACN, Gradient (time / % of B): 0 / 20, 3 / 20, 18 / 40, 18.1 / 100, 20 / 100, 20.1 / 20, 24 / 20, Flow rate: 17ml / min. Fractions containing the desired product were evaporated to give tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-(2-(4-((((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)adamantan-1-yl)carbamate (0.1 g) as an off-white solid.

[0297] Step 3 (2S)-1-(((1S,3R,5S)-3-(2-(2-(4-((((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)adamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (14): [ka]

[0298] To a stirred solution of tert-butyl (2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-(2-(4-((((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)adamantan-1-yl)carbamate (0.1 g, 0.122 mmol) in acetonitrile and water (1:1) mixture (4 mL), 2N aqueous hydrochloric acid solution (1.22 mL, 2.44 mmol) was added and the reaction mixture was stirred at room temperature for 24 h. After completion of the reaction (monitored by LCMS). The reaction mixture was evaporated to obtain the crude product. The obtained crude product was purified by RP preparative HPLC using the following conditions: Column / dimensions: X-BRIDGE C18 (19×250 mm), 5μ, Mobile phase A: 10 mM ABC in water, Mobile phase B: Acetonitrile, Gradient (time / % of B): 0 / 5, 4 / 5, 12 / 75, 12.1 / 5, 15 / 5, Flow rate: 16 ml / min. The desired peak fraction was lyophilized to give (2S)-1-(((1S,3R,5S)-3-(2-(2-(4-((((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)adamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (0.063 g, 73.3%) as an off-white solid. LCMS M / Z 636.49(M+1) 1 H NMR(400 MHz,DMSO-d6)δ 7.92(s,1H),5.10(bs,1H),4.73-4.65(m,2H),4.49-4.43(m,3H),4.3(bs,2H), 3.80-3.36(m,19H),2.64-2.60(m,2H),2.16-2.01(m,6H),1.56-1.46(m,12H). [Example 15] Synthesis of (1S,3S,5S)-2-((2S)-2-amino-2-((1S,3R,5S)-3-(2-hydroxyethoxy)adamantan-1-yl)acetyl)-2-azabicyclo[3.1.0]hexane-3-carbonitrile (16): [ka]

[0299] To a stirred solution of tert-butyl ((1S)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-1-((1S,3R,5S)-3-(2-hydroxyethoxy)adamantan-1-yl)-2-oxoethyl)carbamate (0.30 g, 0.65 mmol), DCM (3 mL), TFA (3 mL) were added dropwise at 0° C. and the reaction mixture was stirred at room temperature for 3 h. After completion of the reaction (monitored by LCMS), the reaction mixture was concentrated, washed with diethyl ether (2×50 mL) and evaporated to obtain the crude product. The crude product obtained was basified with NaHCO3 (20 mL), extracted with DCM (3×50 mL), dried over anhydrous Na2SO4 and concentrated. The crude product was purified by reverse phase C-18 column chromatography using 27% ACN and 10 mM ABC in water. Fractions containing the desired product were collected and lyophilized to give (1S,3S,5S)-2-((2S)-2-amino-2-((1S,3R,5S)-3-(2-hydroxyethoxy)adamantan-1-yl)acetyl)-2-azabicyclo[3.1.0]hexane-3-carbonitrile (0.053 g) as an off-white solid. LCMS M / Z 360.46 (M+1). 1 H NMR(400 MHz,DMSO-d6)δ 5.10-5.07(m,1H),4.46-4.43(m,1H),3.89-3.87(m,1H),3.45-3.31(m,5H ),2.21-2.14(m,3H),1.77-1.32(m,14H),0.97(m,1H),0.72-0.70(s,1H). [Example 16] Synthesis of 2-(((1R,3S,5S)-3-((S)-1-amino-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)adamantan-1-yl)oxy)ethyl(4-((hexyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)phenyl)glycinate (19): [ka]

[0300] To a solution of tert-butyl ((1S)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-1-((1S,3R,5S)-3-(2-hydroxyethoxy)adamantan-1-yl)-2-oxoethyl)carbamate (36 mg, 1.0 equiv.) in dichloromethane (0.21 mL) was added triethylamine (0.04 mL, 4.0 equiv.) and 4-nitrophenyl chloroformate (17.1 mg in 0.13 mL of dichloromethane) at 0° C. After stirring at room temperature for 4 hours, the mixture was added to a solution of (2R,3R,4R,5S)-6-((4-(aminomethyl)benzyl)(hexyl)amino)hexane-1,2,3,4,5-pentaol (46 mg, 1.2 eq.) and triethylamine (0.08 mL, 6.0 eq.) in acetonitrile (0.2 mL+0.05 mL*2 washes) at 0° C. The reaction was stirred for 12 hours, after which saturated aqueous NaHCO3 was added. The mixture was then extracted three times with dichloromethane. The combined organic layers were washed with brine and concentrated in vacuo to give the crude product, which was used in the next step without further purification. The above crude product was treated with trifluoroacetic acid (0.1 mL) and dichloromethane (0.2 mL). After stirring for 4 hours, the volatiles were removed under reduced pressure. The crude product was purified by C18 reverse phase preparative HPLC to give 2-(((1R,3S,5S)-3-((S)-1-amino-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)adamantan-1-yl)oxy)ethyl(4-((hexyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)phenyl)glycinate formate salt (17.4 mg, 25%) as a white powder.ES MS M / Z=501.56(M+1-adamantane)1H NMR(500 MHz,MeOD)δ 7.51(d,J=7.8 Hz,2H),7.42(d,J=7.9 Hz,2H),5.21(dd,J=10.7,2.3 Hz,1H),4.49-4.30(m,4H),4.23-4.12(m,4H),3.94(td,J=6.2,2.7 Hz,1H),3.83-3.75(m,2H),3.73-3.60(m,5H),3.32-3.27(m,2H),3.11(qdd,J=13.0,10.1,6.1 Hz,2H),2.65(ddd,J=13.7,10.6,5.7 Hz,1H),2.39-2.26(m,3H),2.03(dq,J=9.0,5.7 Hz,1H),1.90-1.56(m,16H),1.41-1.23(m,11H),1.14(dt,J=9.0,6.4 Hz,1H),1.00-0.87(m,4H). Similarly, compounds 17 and 18 were synthesized from Int-9.

[0301] [Example 17] Synthesis of 2-(((1R,3S,5S)-3-((S)-1-amino-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)adamantan-1-yl)oxy)ethyl(4-((bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)benzyl)carbamate trifluoroacetate (22): [ka] [ka]

[0302] Step 1 tert-Butyl (4-(azidomethyl)benzyl)carbamate: [ka]

[0303] To a stirred solution of tert-butyl (4-(bromomethyl)benzyl)carbamate (2.0 g, 6.66 mmol) in DMF (20 mL) was added sodium azide (0.86 g, 13.33 mmol) at room temperature and the reaction mixture was stirred at 80° C. for 2 h. After completion, the reaction mixture was quenched with ice-cold water solid, filtered through a Buchner funnel and dried to give tert-butyl (4-(azidomethyl)benzyl)carbamate (1.5 g) as an off-white solid. TLC system: EtOAc:petroleum ether (0.3:0.7); R f :0.4.

[0304] Step 2 tert-Butyl (4-(aminomethyl)benzyl)carbamate: [ka]

[0305] To a stirred solution of tert-butyl (4-(azidomethyl)benzyl)carbamate (1.3 g, 4.95 mmol) in methanol (15 mL) was added triphenylphosphine (1.9 g, 7.43 mmol) and the reaction mixture was stirred at 80° C. for 2 h. After completion, the reaction mixture was evaporated and purified by grace column chromatography using 8-10% MeOH in DCM to give tert-butyl (4-(aminomethyl)benzyl)carbamate (0.9 g) as a gum. TLC system: MeOH:DCM (0.1:0.9); R f :0.2 Step 3 tert-Butyl (4-((bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)benzyl)carbamate: [ka]

[0306] To a stirred solution of tert-butyl (4-(aminomethyl)benzyl)carbamate (0.88 g, 3.72 mmol) and (2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanal (1.6 g, 9.30 mmol) in methanol (26 mL) was added acetic acid (0.8 mL) at 0° C. and the resulting mixture was stirred at room temperature for 20 minutes. Sodium cyanoborohydride (0.49 g, 7.81 mmol) was then added at 0° C. and the reaction mixture was heated at 80° C. for 16 hours. After completion, the reaction mixture was concentrated and purified by RP column chromatography using 25% MeOH and 0.1% FA in water. The pure fractions were evaporated to give tert-butyl (4-((bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)benzyl)carbamate (1.0 g) as a white solid.

[0307] Step 4 (2R,2'R,3R,3'R,4R,4'R,5S,5'S)-6,6'-((4-(aminomethyl)benzyl)azanediyl)bis(hexane-1,2,3,4,5-pentanol) hydrochloride: [ka]

[0308] To a stirred solution of tert-butyl (4-((bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)benzyl)carbamate formate (0.8 g, 1.41 mmol) in methanol (4.0 mL) was added 4 M hydrochloric acid in 1,4-dioxane (8.0 mL) at 0° C. and the mixture was stirred at room temperature for 2 h. After completion of the reaction (monitored by LCMS), the reaction mixture was evaporated to give (2R,2′R,3R,3′R,4R,4′R,5S,5′S)-6,6′-((4-(aminomethyl)benzyl)azanediyl)bis(hexane-1,2,3,4,5-pentaol) hydrochloride (0.95 g, crude) as a white gummy solid.

[0309] Step 5 tert-Butyl ((1S)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-1-((1S,3R,5S)-3-(2-(((4-nitrophenoxy)carbonyl)oxy)oxyethoxy)adamantan-1-yl)-2-oxoethyl)carbamate: [ka]

[0310] To a stirred solution of tert-butyl ((1S)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-1-((1S,3R,5S)-3-(2-hydroxyethoxy)siadamantan-1-yl)-2-oxoethyl)carbamate (0.8 g, 1.74 mmol) in dichloromethane (10 mL) was added triethylamine (0.9 mL, 1.91 mmol) followed by 4-nitrophenyl carbonochloridate (0.38 g, 6.96 mmol) in dichloromethane (2 mL) at -10°C and the reaction mixture was stirred at room temperature for 2 hours. After completion of the reaction, the reaction mixture was taken directly into the next step.

[0311] Step 6. 2-(((1R,3S,5S)-3-((S)-1-((tert-butoxycarbonyl)amino)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)adamantan-1-yl)oxy)ethyl (4-((bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)benzyl)carbamate: [ka]

[0312] To a stirred solution of (2R,2'R,3R,3'R,4R,4'R,5S,5'S)-6,6'-((4-(aminomethyl)benzyl)azanediyl)bis(hexane-1,2,3,4,5-pentanol) hydrochloride (0.96 g, 1.92 mmol) in ACN (10 mL) was added TEA (1.3 mL, 9.60 mmol) at 0 °C. Then tert-butyl ((1S)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-1-((1S,3R,5S)-3-(2-(((4-nitrophenoxy)carbonyl)oxy)ethoxy)adamantan-1-yl)-2-oxoethyl)carbamate (1.17 g, 1.60 mmol) was added dropwise to the reaction mixture at 0° C. and stirred at 80° C. for 16 hours. After completion of the reaction, the reaction mixture was concentrated and purified by RP C18 column chromatography using 25% MeOH and 0.1M FA in water. Fractions containing the desired product were collected and evaporated to give 2-(((1R,3S,5S)-3-((S)-1-((tert-butoxycarbonyl)amino)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)adamantan-1-yl)oxy)ethyl (4-((bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)benzyl)carbamate (0.8 g) as a yellow gum.

[0313] Step 7. 2-(((1R,3S,5S)-3-((S)-1-amino-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)adamantan-1-yl)oxy)ethyl(4-((bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)benzyl)carbamate trifluoroacetate (22): [ka]

[0314] To a stirred solution of 2-(((1R,3S,5S)-3-((S)-1-((tert-butoxycarbonyl)amino)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)adamantan-1-yl)oxy)ethyl (4-((bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)benzyl)carbamate (0.80 g, 0.84 mmol), DCM (8 mL), TFA (4 mL) was added dropwise at 0° C. and the reaction mixture was stirred at room temperature for 1 h. After completion of the reaction (monitored by LCMS), the reaction mixture was concentrated, washed with diethyl ether (2×100 mL) and evaporated to give the crude product. The resulting crude product was purified by RP SUNFIRE-C18 (150*19*5μ) using 20% ​​ACN and 0.1% TFA in water. Fractions containing the desired product were collected and lyophilized to give 2-(((1R,3S,5S)-3-((S)-1-amino)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)adamantan-1-yl)oxy)ethyl(4-((bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)benzyl)carbamate trifluoroacetate (0.184 g) as an off-white solid. LCMS M / Z 850.68(M+1) 1 H NMR(400 MHz,DMSO-d6)δ 8.54(s,1H),8.17(s,2H),7.79(t,J=6.0 Hz,1H),7.48(d,J=8.0 Hz,2H),7.32(d,J=8.0 Hz,2H),5.60(bs,1H),4.95(bs,1H),5.27-5.24(m,1H),4.95(m,2H),4.37-4.02(m,16H),3.72-3.52 (m,12H),3.16(m,4H),2.32-2.23(m,3H),2.07(m,1H),1.73-1.46(m,12H),1.04(m,1H),0.75(s,1H). [Example 18] Synthesis of 2-(((1R,3S,5S)-3-((S)-1-amino-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)adamantan-1-yl)oxy)ethyl(4-((ethyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)benzyl)carbamate formate (23): [ka]

[0315] Step 1 tert-Butyl ((1S)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-1-((1S,3R,5S)-3-(2-(((4-nitrophenoxy)carbonyl)oxy)oxyethoxy)adamantan-1-yl)-2-oxoethyl)carbamate: [ka]

[0316] To a stirred solution of tert-butyl ((1S)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-1-((1S,3R,5S)-3-(2-hydroxyethoxy)adamantan-1-yl)-2-oxoethyl)carbamate (0.8 g, 1.74 mmol) in DCM (10 mL) was added TEA (0.9 mL, 6.96 mmol) followed by dropwise addition of 4-nitrophenyl carbonochloridate (0.38 g, 1.91 mmol) in DCM (0.5 mL) at -10 °C and stirred at room temperature for 2 h. After completion, the reaction mixture was taken to the next step to give tert-butyl ((1S)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-1-((1S,3R,5S)-3-(2-(((4-nitrophenoxy)carbonyl)oxy)oxyethoxy)adamantan-1-yl)-2-oxoethyl)carbamate (1.17 g) as a yellow solution. TLC system: 100% EtOAc, R f :0.4 Step 2. 2-(((1R,3S,5S)-3-((S)-1-((tert-butoxycarbonyl)amino)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)adamantan-1-yl)oxy)ethyl(4-((ethyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)benzyl)carbamate: [ka]

[0317] To a stirred solution of tert-butyl (2R,3R,4R,5S)-6-((4-(aminomethyl)benzyl)(ethyl)amino)hexane-1,2,3,4,5-pentanol hydrochloride (0.81 g, 2.24 mmol) in ACN (10 mL) was added TEA (1.5 mL, 11.22) at 0° C. Then tert-butyl ((1S)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-1-((1S,3R,5S)-3-(2-(((4-nitrophenoxy)carbonyl)oxy)ethoxy)adamantan-1-yl)-2-oxoethyl)carbamate (1.17 g, 1.87 mmol) was added dropwise to the reaction mixture at 0° C. and stirred at 80° C. for 16 h. After completion of the reaction, the reaction mixture was concentrated and purified by RP C18 column chromatography using 27% ACN and 0.1M FA in water. The fractions containing the desired product were collected and evaporated to give 2-(((1R,3S,5S)-3-((S)-1-((tert-butoxycarbonyl)amino)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)adamantan-1-yl)oxy)ethyl(4-((ethyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)benzyl)carbamate (1.0 g) as a yellow gum.

[0318] Step 3. 2-(((1R,3S,5S)-3-((S)-1-amino-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)adamantan-1-yl)oxy)ethyl(4-((ethyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)benzyl)carbamate formate (23): [ka]

[0319] To a stirred solution of 2-(((1R,3S,5S)-3-((S)-1-((tert-butoxycarbonyl)amino)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)adamantan-1-yl)oxy)ethyl (4-((ethyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)benzyl)carbamate (1 g, 1.22 mmol) in DCM (10 mL) was added TFA (10 mL) dropwise at 0° C. and the resulting mixture was stirred at room temperature for 1 h. After completion (monitored by LCMS), the reaction mixture was concentrated and washed with diethyl ether (2×100 mL) to give the crude product. The resulting crude product was then purified by RP chromatography using a C-18 column and 0.1M FA in 20% CAN:water as eluent. Fractions containing the desired product were collected and lyophilized to give 2-(((1R,3S,5S)-3-((S)-1-amino)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexan-2-yl)-2-oxoethyl)adamantan-1-yl)oxy)ethyl(4-((ethyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)benzyl)carbamate formate (0.59 g) as an off-white solid. LCMS M / Z 712.50 (M-1). 1H NMR(400 MHz,DMSO-d6)δ 8.13(s,1H),7.74(t,J=6.0 Hz,1H),7.38(d,J=7.6 Hz,2H),7.24(d,J=7.6 Hz,2H),5.22(m,1H),4.18-4.02(m,10H),3.89(s,1H),2.17-3.66-3.52(m,4H),3.49-3.47(m,4H),2.92-2. 71(m,4H),2.50(s,3H),2.27-2.21(m,3H),1.96(m,1H),1.71-1.43(m,12H),1.09-1.02(m,4H),0.74(s,1H). [Example 21] Synthesis of (R)-(2-((8-(3-aminopiperidin-1-yl)-7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)phenyl)boronic acid (27): [ka]

[0320] Step 1 (R)-(2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)phenyl)boronic acid: [ka]

[0321] To a stirred solution of tert-butyl (R)-(1-(7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-8-yl)piperidin-3-yl)carbamate (0.1 g, 0.24 mmol) and (2-(bromomethyl)phenyl)boronic acid (0.08 g, 0.36 mmol) in DMF (1 mL) was added K2CO3 (0.03 g, 0.24 mmol) and the reaction mixture was stirred at room temperature for 16 hours. After completion of the starting material, the reaction mixture was diluted with EtOAc (20 mL), washed with water (20 mL) and brine solution (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain crude mass. The crude residue was purified by reverse phase preparative HPLC to give 0.06 g of (R)-(2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)phenyl)boronic acid as an off-white solid. [TLC system: EtOAc:petroleum ether (8:2); Rf value: 0.5].

[0322] Step 2 (R)-(2-((8-(3-aminopiperidin-1-yl)-7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)phenyl)boronic acid (27): [ka]

[0323] To a stirred solution of (R)-(2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)phenyl)boronic acid (0.06 g, 0.11 mmol) in 1,4-dioxane (0.6 mL) was added dropwise 4M HCl in 1,4-dioxane (0.12 mL) at 0° C. and the reaction mixture was stirred at room temperature for 3 hours. After completion of the starting material, the reaction mixture was concentrated under reduced pressure and triturated with diethyl ether to give 0.025 g of compound 27 (HCl salt) as an off-white solid. [TLC system: MeOH:DCM (1:9); Rf value: 0.2]. LC MS M / Z=451.46 (M+1). 1 H NMR(400 MHz,DMSO-d6)δ 8.22(s,2H),8.12(s,2H),7.55(d,J=6.8 Hz,1H),7.27-7.14(m,2H),6.91(d,J=7.6 Hz,1H),5.25(s,2H),5.00-4.88(m,2H),3.69(d,J=2.4 Hz,1H),3.49-3.40(m,5H),3.22-3.14(m,2H),2.00-1.90(m,2H),1.79(s,3H),1.71-1.61(m,2H). [Example 22] Synthesis of hexyl (R)-2-((8-(3-aminopiperidin-1-yl)-7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-fluoronicotinate (30): [ka]

[0324] Step 1 (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-fluoronicotinic acid [ka]

[0325] To a stirred solution of methyl (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-fluoronicotinate (0.9 g, 1.56 mmol) in THF (9 mL) and water (2 mL) was added LiOH.H2O (0.129 g, 3.08 mmol) at 0° C. The reaction mixture was stirred at room temperature for 4 hours. After completion of the starting material, the reaction mixture was concentrated under reduced pressure to obtain the crude compound. The crude compound was diluted with water and acidified with 1N HCl. The precipitated solid was filtered off and dried under vacuum to give 0.72 g of the compound (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-fluoronicotinic acid as an off-white solid. [TLC system: MeOH:DCM (1:9); R f Value: 0.3].

[0326] Step 2 Hexyl (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-fluoronicotinate: [ka]

[0327] To a stirred solution of the compound (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-fluoronicotinic acid (0.72 g, 1.26 mmol) in DMF (15 mL) was added CH 13Br (0.25 g, 1.51 mmol) and K2CO3 (0.52 g, 3.78 mmol) were added and the reaction mixture was stirred at room temperature for 16 hours. After completion of the starting material, the reaction mixture was quenched with cold water and the precipitated solid was filtered off and dried under vacuum to give 0.3 g of compound hexyl (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-fluoronicotinate as an off-white solid. [TLC system: EtOAc:petroleum ether (1:1); R f Value: 0.8].

[0328] Step 3 Hexyl (R)-2-((8-(3-aminopiperidin-1-yl)-7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-fluoronicotinate (30): [ka]

[0329] To a stirred solution of compound hexyl (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-fluoronicotinate (0.30 g, 0.46 mmol) in 1,4-dioxane (3 mL), 4M HCl in 1,4-dioxane (2 mL) was added at 0° C. and the reaction mixture was stirred at room temperature for 2 hours. After completion of the starting material, the reaction mixture was concentrated under reduced pressure to obtain crude compound. The crude compound was purified by reverse phase preparative HPLC to obtain 0.181 g of the compound hexyl (R)-2-((8-(3-aminopiperidin-1-yl)-7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-fluoronicotinate as an off-white solid. [TLC system: MeOH:DCM (1:9); Rf Value: 0.3]. LC MS M / Z=554.87(M+1) 1 H NMR(400 MHz,DMSO-d6)δ 8.48(t,J=8.4 Hz,1H),8.12(d,J=3.6 Hz,3H),7.22-7.19(dd,J=8.4 Hz,2.4 Hz,1H),5.49(s,2H),4.94(m,2H),4.34(t,J=6.4 Hz,2H),3.71(m,1H),3.50-3.40(m,5H),3.21-3.16(m,2H),2.02-1.89(m,1H),1.89-1. 77(m,1H),1.76-1.65(m,7H),1.44-1.41(m,2H),1.34-1.31(m,4H),0.89-0.86(m,3H). [Example 23] Synthesis of hexyl (R)-2-((8-(3-aminopiperidin-1-yl)-7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-(methylamino)nicotinic acid (31): [ka]

[0330] To a stirred solution of compound (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-(methylamino)nicotinic acid (0.5 g, 0.86 mmol) in 1,4-dioxane (3 mL), 4M HCl in 1,4-dioxane (2 mL) was added at 0° C. and the reaction mixture was stirred at room temperature for 2 hours. After completion of the starting material, the reaction mixture was concentrated under reduced pressure to obtain crude compound. The crude compound was purified by reverse phase preparative HPLC to obtain 0.157 g of compound hexyl(R)-2-((8-(3-aminopiperidin-1-yl)-7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-(methylamino)nicotinic acid as an off-white solid. [TLC system: MeOH:DCM (1:9); R f Value: 0.3]. LC MS M / Z=481.28(M+1) 1 H NMR(400 MHz,DMSO-d6)δ 8.24(m,3H),7.87(d,J=7.6 Hz,1H),6.33(d,J=8.4 Hz,1H),5.37(s,2H),5.00-4.88(m,2H),3.68-3.38(m,6H),3.18-3.12( m,2H),2.45(m,3H),2.02-1.90(m,2H),1.77(s,3H),1.70-1.62(m,2H). [Example 24] Synthesis of hexyl (R)-2-((8-(3-aminopiperidin-1-yl)-7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-(methylamino)nicotinate (32): [ka]

[0331] Step 1 (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-(methylamino)nicotinic acid: [ka]

[0332] To a stirred solution of the compound (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-(methylamino)nicotinic acid (1.0 g, 1.72 mmol, shares the same intermediate as mCMT478) in DMF (10 mL) was added CH 13 Br (0.34 g, 2.06 mmol) and K2CO3 (0.71 g, 5.16 mmol) were added and the reaction mixture was stirred at room temperature for 16 hours. After completion of the starting material, the reaction mixture was quenched with cold water and the precipitated solid was filtered off and dried under vacuum to give 0.7 g of compound (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-(methylamino)nicotinic acid as an off-white solid. [TLC system: MeOH:DCM (1:9); R f Value: 0.8].

[0333] Step 2. Hexyl (R)-2-((8-(3-aminopiperidin-1-yl)-7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-(methylamino)nicotinate (32): [ka]

[0334] To a stirred solution of compound hexyl (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-(methylamino)nicotinate (0.41 g, 0.62 mmol) in 1,4-dioxane (4 mL), 4M HCl in 1,4-dioxane (3 mL) was added at 0° C. and the reaction mixture was stirred at room temperature for 2 hours. After completion of the starting material, the reaction mixture was concentrated under reduced pressure to obtain crude compound. The crude compound was purified by reverse phase preparative HPLC to obtain 0.161 g of compound hexyl (R)-2-((8-(3-aminopiperidin-1-yl)-7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-(methylamino)nicotinate as an off-white solid. [TLC system: MeOH:DCM (1:9); R f Value: 0.3]. LC MS M / Z=565.79(M+1) 1 H NMR(400 MHz,DMSO-d6)δ 8.18(s,3H),7.87(d,J=8.4 Hz,1H),7.21(br s,1H),6.34(d,J=8.8,1H),5.36(s,2H),4.99-4.88(m,2H),4.22(t,J=6.4 Hz,2H),3.70-3.59(m,1H),3.49-3.39(m,5H),3.18-3.11(m,2H),2.44(s, 3H),2.02-1.99(m,2H),1.90-1.61(m,7H),1.42-1.31(m,6H),0.89(s,3H). [Example 25] Synthesis of methyl 2-((8-((R)-3-aminopiperidin-1-yl)-7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-((7-(4-((bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)heptyl)amino)nicotinate (34): [ka]

[0335] To a solution of compound methyl 6-((7-(4-((bis((2S,3R)-2,3-dihydroxy-3-((4R,5R)-5-hydroxy-2-phenyl-1,3-dioxan-4-yl)propyl)amino)methyl)-1H-1,2,3-triazol-1-yl)heptyl)amino)-2-((7-(but-2-yn-1-yl)-8-((R)-3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)nicotinate (0.9 g, 0.70 mmol) in HO (10 mL) was added concentrated HCl (2 mL) at 0° C. and the reaction mixture was stirred at room temperature for 2 h. After completion of the reaction (LCMS monitoring), the reaction mixture was concentrated under reduced pressure to obtain crude mass. The crude product was purified by RP-HPLC to obtain compound methyl 2-((8-((R)-3-aminopiperidin-1-yl)-7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-((7-(4-((bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)heptyl)amino)nicotinate (0.041 g) as an off-white solid. LC MS M / Z=1003.62(M+1) 1 H NMR(400 MHz,DMSO d6)δ 8.95(s,1H),8.34(s,1H),8.21(m,3H),7.84(d,J=8.7 Hz,1H),7.39(brs,1H),6.31(d,J=8.8 Hz,1H),5.34(br m,4H),4.93(m,2H),4.56(m,2H),4.39(t,J=6.9 Hz,2H),4.13(m,2H),3.79(s,3H),3.59(m,6H),3.43(m,11H),3.15(m,6H),2 .86(s,2H),2.04(m,1H),1.91(m,1H),1.78(m,7H),1.16(m,8H),0.89(m,2H). [Example 26] Synthesis of 2-((8-((R)-3-aminopiperidin-1-yl)-7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-((7-(4-((bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)heptyl)amino)nicotinic acid (35): [ka]

[0336] Step 1. 6-((7-(4-((bis((2S,3R)-2,3-dihydroxy-3-((4R,5R)-5-hydroxy-2-phenyl-1,3-dioxan-4-yl)propyl)amino)methyl)-1H-1,2,3-triazol-1-yl)heptyl)amino)-2-((7-(but-2-yn-1-yl)-8-((R)-3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)nicotinic acid: [ka]

[0337] To a solution of compound methyl 6-((7-(4-((bis((2S,3R)-2,3-dihydroxy-3-((4R,5R)-5-hydroxy-2-phenyl-1,3-dioxan-4-yl)propyl)amino)methyl)-1H-1,2,3-triazol-1-yl)heptyl)amino)-2-((7-(but-2-yn-1-yl)-8-((R)-3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)nicotinate (0.350 g, 0.27 mmol) in MeOH (5 mL) and HO (1 mL), NaOH (0.054 g, 1.35 mmol) was added and the reaction mixture was stirred at room temperature for 16 h. After completion of the reaction (LCMS monitoring). The reaction mixture was concentrated under reduced pressure to obtain crude mass. The crude compound was washed with Et2O (50 mL) and dried in vacuum to obtain compound 6-((7-(4-((bis((2S,3R)-2,3-dihydroxy-3-((4R,5R)-5-hydroxy-2-phenyl-1,3-dioxan-4-yl)propyl)amino)methyl)-1H-1,2,3-triazol-1-yl)heptyl)amino)-2-((7-(but-2-yn-1-yl)-8-((R)-3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)nicotinic acid (0.3 g) as an off-white solid. The crude compound was taken to the next step.

[0338] Step 2. 2-((8-((R)-3-aminopiperidin-1-yl)-7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-((7-(4-((bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)heptyl)amino)nicotinic acid (35): [ka]

[0339] To a solution of compound 6-((7-(4-((bis((2S,3R)-2,3-dihydroxy-3-((4R,5R)-5-hydroxy-2-phenyl-1,3-dioxan-4-yl)propyl)amino)methyl)-1H-1,2,3-triazol-1-yl)heptyl)amino)-2-((7-(but-2-yn-1-yl)-8-((R)-3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)nicotinic acid (0.3 g, 0.24 mmol) in HO (10 mL) was added concentrated HCl (0.6 mL) at 0° C. and the reaction mixture was stirred at room temperature for 2 h. After completion of the reaction (LCMS monitoring), the reaction mixture was concentrated under reduced pressure to obtain crude mass. The crude product was purified by RP preparative HPLC to obtain compound 2-((8-((R)-3-aminopiperidin-1-yl)-7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-6-((7-(4-((bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)heptyl)amino)nicotinic acid (0.014 g) as an off-white solid. LC MS M / Z=987.62(M+1) 1 H NMR(400 MHz,DMSO)δ 7.93(s,1H),7.79(d,J=8.6 Hz,1H),7.01(br s,1H),6.22(d,J=8.6 Hz,1H),5.31(s,2H),4.59-4.26(m,14H),3.77(m,4H),3.55(m,9H),3.38(m,7H) ,2.90(m,5H),2.50(m,2H),1.74(m,9H),1.27(m,1H),1.12(m,6H),0.91(m,2H). [Example 27] Synthesis of nonyl (R)-2-((8-(3-aminopiperidin-1-yl)-7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoate (37): [ka]

[0340] Step 1 Isopropyl (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoate: [ka]

[0341] To a stirred solution of compound (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoic acid (0.5 g, 0.85 mmol) in DMF (5 mL), 2-bromopropane (0.15 g, 1.28 mmol) and K2CO3 (0.35 g, 2.56 mmol) were added and the reaction mixture was stirred at room temperature for 16 hours. After completion of the starting material, the reaction mixture was quenched with cold water. The precipitated solid was filtered off, dried under vacuum and washed with diethyl ether to obtain 0.4 g of compound isopropyl (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoate as an off-white solid. [TLC system: EtOAc:petroleum ether (3:7); R f Value: 0.7].

[0342] Step 2. Synthesis of isopropyl (R)-2-((8-(3-aminopiperidin-1-yl)-7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoate (37): [ka]

[0343] To a stirred solution of compound isopropyl (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoate (0.4 g, 0.64 mmol) in 1,4-dioxane (4 mL), 4M HCl in 1,4-dioxane (4 mL) was added at 0° C. and the reaction mixture was stirred at room temperature for 3 hours. After completion of starting material, the reaction mixture was concentrated under reduced pressure to obtain crude mass. The crude compound was purified by reverse phase preparative HPLC to obtain 0.19 g of compound isopropyl (R)-2-((8-(3-aminopiperidin-1-yl)-7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoate as an off-white solid. [TLC system: MeOH:DCM (1:9); R f Value: 0.2]. LC MS M / Z=527.49(M+1) 1 H NMR(400 MHz,DMSO-d6)δ 7.83(d,J=2.0 Hz,1H),7.54(dd,J=8.4,2.0 Hz,1H),7.08(d,J=8.4 Hz,1H),5.32(s,2H),5.22-5.16(m,1H),4.88(s,2H),3.66-3.58(m,2H),3.38(s,3H),2.99(t,J=10.4 Hz,1H),2.84-2.72(m,2H),1.87-1.57(m,8H),1.37(d,J=6.0 Hz,6H),1.27-1.26(m,1H). Similarly, hexyl (R)-2-((8-(3-aminopiperidin-1-yl)-7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoate (38) was synthesized.

[0344] [Example 28] Synthesis of nonyl (R)-2-((8-(3-aminopiperidin-1-yl)-7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoate (39): [ka]

[0345] Step 1 Nonyl (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoate: [ka]

[0346] To a stirred solution of (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoic acid (0.35 g, 0.6 mmol) in DMF (3.5 mL) was added 1-bromononane (0.15 g, 0.72 mmol) and K2CO3 (0.25 g, 1.8 mmol) and the reaction mixture was stirred at room temperature for 16 hours. After completion of the starting material, the reaction mixture was quenched with cold water. The precipitated solid was filtered off, dried under vacuum, and washed with diethyl ether to obtain 0.25 g of nonyl (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoate as an off-white solid. [TLC system: EtOAc:petroleum ether (1:1); Rf value: 0.7].

[0347] Step 2. Synthesis of nonyl (R)-2-((8-(3-aminopiperidin-1-yl)-7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoate (39): [ka]

[0348] To a stirred solution of nonyl (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoate (0.35 g, 0.49 mmol) in 1,4-dioxane (2 mL) was added 4M HCl in 1,4-dioxane (2 mL) at 0° C. and the reaction mixture was stirred at room temperature for 3 hours. After completion of starting material, the reaction mixture was concentrated under reduced pressure to obtain crude mass. The crude compound was purified by reverse phase preparative HPLC to give 0.13 g of nonyl (R)-2-((8-(3-aminopiperidin-1-yl)-7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoate (39) as a brown solid. [TLC system: MeOH:DCM (1:9); Rf value: 0.3]. LC MS M / Z=611.64 (M+1). 1 H NMR(400 MHz,DMSO-d6)δ 7.85(d,J=2.4 Hz,1H),7.55(dd,J=8.4,2.0 Hz,1H),7.08(d,J=8.4 Hz,1H),5.32(s,2H),4.88(s,2H),4.32(t,J=6.4 Hz,2H),3.66-3.58(m,2H),3.37(s,3H),3.01-2.96(m,1H),2.82-2.72(m,2H),1.87-1.60(m,10H),1.41-1.24(m,13H),0.84(t,J=6.4 Hz,3H). Similarly, (R)-2-((8-(3-aminopiperidin-1-yl)-7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chloro-N-pentylbenzamide (40) was synthesized.

[0349] [Example 29] Synthesis of 2-(2-(4-((hexyl(2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3 triazol-1-yl)ethoxy)ethyl 2-((8-((R)-3-aminopiperidin-1-yl)-7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoate (41): [ka]

[0350] Step 1. 2-(2-Bromoethoxy)ethyl (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoate: [ka]

[0351] To a stirred solution of (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoic acid (1.0 g, 1.71 mmol) and 1-bromo-2-(2-bromoethoxy)ethane (1.98 g, 8.55 mmol) in DMF (10 mL) was added KCO (0.71 g, 5.13 mmol) and the reaction mixture was stirred at room temperature for 16 h. After completion of the starting material, the reaction mixture was quenched with cold water and the precipitated solid was filtered off and dried under vacuum to give 1.0 g of 2-(2-bromoethoxy)ethyl (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoate as an off-white solid. [TLC system: EtOAc:petroleum ether (3:7); R f Value: 0.6].

[0352] Step 2. 2-(2-azidoethoxy)ethyl (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoate: [ka]

[0353] To a stirred solution of 2-(2-bromoethoxy)ethyl (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoate (3) (1.0 g, 1.36 mmol) in DMF (10 mL) was added NaN3 (0.44 g, 6.79 mmol) and the reaction mixture was stirred at room temperature for 16 hours. After completion of the starting material, the reaction mixture was quenched with water and extracted with ethyl acetate (3X 25 mL). The combined organic layers were washed with brine solution (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the crude compound. The crude compound was purified by column chromatography (silica gel 100-200 mesh, 30% ethyl acetate in petroleum ether used as eluent) to give 0.7 g of 2-(2-azidoethoxy)ethyl (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoate as an off-white solid. [TLC system: EtOAc:petroleum ether (1:1); Rf value: 0.4].

[0354] Step 3. 2-(2-(4-((hexyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethyl 2-((7-(but-2-yn-1-yl)-8-((R)-3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoate: [ka]

[0355] 2-(2-Azidoethoxy)ethyl (R)-2-((7-(but-2-yn-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoate (0. To a stirred solution of (2R,3R,4R,5S)-6-(hexyl(prop-2-yn-1-yl)amino)hexane-1,2,3,4,5-pentaol 5 (0.26 g, 0.86 mmol) was added CuSO4.5H2O (0.13 g, 0.52 mmol) and sodium ascorbate (0.13 g, 0.65 mmol). The reaction mixture was stirred at room temperature for 8 h. After completion of the starting material, the reaction mixture was quenched with water and extracted with DCM (3 x 20 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give 0.45 g of crude 2-(2-(4-((hexyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethyl 2-((7-(but-2-yn-1-yl)-8-((R)-3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoate as a yellow solid. [TLC system: MeOH:DCM (1:9); Rf value: 0.5]. This crude compound was used in the next step without further purification.

[0356] Step 4. 2-(2-(4-((hexyl(2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3 triazol-1-yl)ethoxy)ethyl 2-((8-((R)-3-aminopiperidin-1-yl)-7-(but-2-yn-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoate (41): [ka]

[0357] To a stirred solution of 2-(2-(4-((hexyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethyl 2-((7-(but-2-yn-1-yl)-8-((R)-3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1-yl)methyl)-5-chlorobenzoate (0.45 g, 0.45 mmol) in 1,4-dioxane (4.5 mL) was added 4M HCl in 1,4-dioxane (1 mL) dropwise at 0° C. and the reaction mixture was stirred at room temperature for 2 hours. After completion of the starting material, the reaction mixture was concentrated under reduced pressure to give the crude compound. The crude compound was purified by reverse phase preparative HPLC to give 0.1 g of compound 41 (HCL salt) as an off-white solid. [TLC system: MeOH:DCM (1:9); f Value: 0.2]. LCMS M / Z=901.25(M+H) 1 H NMR(400 MHz,DMSO-d6)δ 9.5(s,1H),8.31(d,J=7.6 Hz,1H),8.23(s,3H),7.84(s,1H),7.57(dd,J=8.4,2.0 Hz,1H),7.08(d,J=8.4 Hz,1H),5.33(s,3H),5.01-4.87(m,2H),4.63(s,2H),4.41(s,4H),4.12-4.03(m,1H),3.93-3.91(m,2H),3.79(m,2H),3.74-3. 60(m,3H),3.39(m,8H),3.20-3.17(m,4H),2.99-2.98(m,2H),2.02-1.91(m,2H),1.78-1.63(m,7H),1.25(s,6H),0.86(s,3H). [Example 30] 2-(2-(2-azidoethoxy)ethoxy)ethyl (R)-7-(3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (42): [ka]

[0358] To a solution of the compound 2-(2-(2-azidoethoxy)ethoxy)ethyl (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (0.4 g, 0.565 mmol) in DCM (10 mL) was added 4M HCl in 1,4-dioxane at 0° C. The reaction was then stirred at room temperature for 3 h. After completion of the reaction, the reaction mixture was concentrated and the residue was basified with saturated NaHCO3 solution, extracted with EtOAc (20 mL x 2), dried over anhydrous sodium sulfate and evaporated to give the crude product, which was purified by column chromatography on silica gel (Davisil) using 0-8% MeOH in DCM as eluent to give 30 mg of 2-(2-(2-azidoethoxy)ethoxy)ethyl (R)-7-(3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate as a light brown gum. [TLC system: MeOH:DCM (1:9); R f Value: 0.5]. LC MS M / Z=608.64(M+1). 1 H NMR(400 MHz,DMSO-d6)δ 7.45-7.43(m,2H),4.96-4.93(m,2H),4.37-4.36(m,2H),4.25-4.11(m,2H),3.94-3.93(m,2H),3.75-3.71(m ,2H),3.60-3.58(m,6H),3.37-3.34(m,2H),3.31(m,1H),2.73-2.58(m,2H),2.51-2.49(m,2H),1.80(bs,2H). [Example 31] Synthesis of 2-(2-(2-(4-((((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (44): [ka]

[0359] Step 1. 2-(2-(2-(4-((((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate: [ka]

[0360] To a stirred solution of the compound 2-(2-(2-azidoethoxy)ethoxy)ethyl (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (0.6 g, 0.847 mmol) and the compound 6-(prop-2-yn-1-ylamino)hexane-1,2,3,4,5-pentanol (0.371 g, 1.695 mmol) in t-BuOH (5 mL) and water (1 mL) was added CuSO 4,5. H2O (0.254 g, 1.017 mmol) and Na ascorbate (0.251 g, 1.271 mmol) were added. The resulting mixture was stirred at room temperature for 16 hours. After completion of the reaction, the reaction mixture was filtered. The filtrate was concentrated under reduced pressure to give the crude compound. The crude compound was purified by RP preparative HPLC to obtain 60 mg of compound 2-(2-(2-(4-((((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-((tert-butoxycarbonyl)amino)-4-(2,4,5 trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate as a brown sticky solid.

[0361] Preparative HPLC purification method: Column / dimensions: X-bridge phenyl (19*250*5μ) Mobile phase A: 10 Mm ammonium bicarbonate in water Mobile phase B: Acetonitrile Gradient (% of time / B): 0 / 10, 1 / 10, 27 / 75, 27.10 / 95, 29.10 / 95, 31.20 / 10, 35 / 10 Flow rate: 18ml / min. Solubility: ACN+water Step 2. 2-(2-(2-(4-((((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (44): [ka]

[0362] To a solution of compound 2-(2-(2-(4-((((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-((tert-butoxycarbonyl)amino)-4-(2,4,5 trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (0.060 g, 0.064 mmol) in DCM (1 mL) was added 4M HCl in 1,4-dioxane (0.6 mL). The reaction mixture was stirred at room temperature for 3 hours. After completion of the reaction, the reaction mixture was filtered and concentrated to obtain crude compound. The crude compound was purified by RP prep HPLC to give 0.0157 g of 2-(2-(2-(4-((((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate as an off-white solid. LC MS M / Z=827.41(M+1) 1 H NMR(400 MHz,DMSO-d6)δ 7.89(s,1H),7.45-7.41(m,2H),5.10-4.85(m,3H),4.60-4.10(m,10H),4. 00-3.30(m,20H),3.10-3.05(m,1H),2.71-2.50(m,5H),2.10-1.40(m,2H). Preparative HPLC purification method: Column / dimensions: X BRIDGE C18 (19mm*250mm*5μ) Mobile phase A: 10mM ABC in water Mobile phase B: Acetonitrile Gradient (% of time / B): 0 / 10, 1 / 10, 18 / 44, 18.10 / 95, 20.10 / 95, 20.20 / 10, 22 / 10 [Example 32] Synthesis of 2-(2-(2-(4-((ethyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (45): [ka]

[0363] Step 1. 6-(Ethylamino)hexane-1,2,3,4,5-pentanol: [ka]

[0364] To a solution of compound 2,3,4,5,6-pentahydroxyhexanal (5.0 g, 27.75 mmol) and compound ethanamine (20.8 mL, 41.62 mmol) in MeOH (30 mL) was added Raney Ni (5.0 g). The reaction mixture was stirred at 60° C. under H2 atmosphere (150 psi) for 16 h. After completion of the reaction, the reaction mixture was filtered through a bed of Celite and the filtrate was concentrated to give compound 6-(ethylamino)hexane-1,2,3,4,5-pentaol (crude) 5.0 g as an off-white solid, which was used in the next step without purification.

[0365] Step 2. 6-(Ethyl(prop-2-yn-1-yl)amino)hexane-1,2,3,4,5-pentanol: [ka]

[0366] To a solution of compound 6-(ethylamino)hexane-1,2,3,4,5-pentaol (2.0 g, 9.563 mmol), compound 3-bromoprop-1-yne (0.85 mL, 11.47 mmol) in THF (30 mL) was added and the reaction mixture was heated at 60° C. for 16 hours. After completion of the reaction, the reaction mixture was filtered and the filtrate was concentrated to obtain 0.450 g of compound 6-(ethyl(prop-2-yn-1-yl)amino)hexane-1,2,3,4,5-pentaol as a brown viscous compound, which was used in the next step without purification.

[0367] Step 3. 2-(2-(2-(4-((ethyl((2S,3R,4R,5R)-(2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate: [ka]

[0368] To a solution of the compound 2-(2-(2-azidoethoxy)ethoxy)ethyl (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (0.6 g, 0.847 mmol) and the compound 6-(ethyl(prop-2-yn-1-yl)amino)hexane-1,2,3,4,5-pentanol (0.420 g, 169 mmol) in t-BuOH (10 mL) and water (2 mL) was added CuSO 4,5. H2O (0.254 g, 1.017 mmol) and Na ascorbate (0.251 g, 1.271 mmol) were added. The resulting reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction, the reaction mixture was filtered and concentrated to obtain the crude compound. The crude compound was purified by preparative HPLC to obtain 0.240 g of compound 2-(2-(2-(4-((ethyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-((tert-butoxycarbonyl)amino)-4-(2,4,5 trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate as a brown viscous compound.

[0369] Preparative HPLC method: Column / Dimensions: X BRIDGE C8(19*250*5um) Mobile phase A: 10MM ABC in water Mobile phase B: Acetonitrile (org) Gradient (% of time / B): 0 / 20, 1 / 20, 9 / 50, 14 / 50, 14.1 / 98, 18 / 98, 18.1 / 20, 21 / 20 Flow rate: 17ml / min, Solubility: acetonitrile + THF + water Step 4. Synthesis of 2-(2-(2-(4-((ethyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-amino-4-(2,4,5 trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (45): [ka]

[0370] To a solution of compound 2-(2-(2-(4-((ethyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-((tert-butoxycarbonyl)amino)-4-(2,4,5 trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (0.240 g, 0.280 mmol) in DCM (3 mL) was added 4M HCl in 1,4-dioxane (2.4 mL). The reaction mixture was stirred at room temperature for 2 hours. After completion of the reaction, the reaction mixture was filtered and concentrated under reduced pressure to obtain crude compound. The crude compound was purified by trituration with diethyl ether to give 175 mg of 2-(2-(2-(4-((ethyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-amino-4-(2,4,5 trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate as an off-white solid. LC MS M / Z=855.65(M+1) 1 H NMR(400 MHz,DMSO-d6)δ 9.55(s,1H),8.29(d,J=5.8 Hz,1H),8.10-8.00(m,3H),7.58(q,J=9.0 Hz,2H),5.49(s,1H),4.95-4.93(m,3H),4.57-3.38(m,28H),3.18-2.66(m,8H),1.31-1.26(m,3H). [Example 33] Synthesis of 2-(2-(2-(4-((hexyl(2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (46) [ka]

[0371] Step 1. 2-(2-(2-(4-((hexyl(2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate: [ka]

[0372] A solution of 2-(2-(2-azidoethoxy)ethoxy)ethyl (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (460 mg, 0.65 mmol) in THF (3 mL). To the reaction mixture was added 2R,3R,4R,5S)-6-(hexyl(prop-2-yn-1-yl)amino)hexane-1,2,3,4,5-pentaol (prepared in step 4 of Example 9) (295 mg, 0.97 mmol), sodium ascorbate (193 mg, 0.97 mmol), CuSO4·5H2O (242.5 mg, 0.97 mmol) and H2O (2 mL). The mixture was stirred at room temperature for 19 h. The reaction mixture was filtered and the filtrate was purified by preparative HPLC (Gemini-C18 150×21.2 mm, 5 um, mobile phase: ACN (0.1% FA)-HO (0.1% FA), gradient: 20-70%) to give 2-(2-(2-(4-((hexyl(2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (140 mg, 21.2%) as a yellow solid. MS(ESI): mass calcd.for C 44 H 64 F6N8O 12 1010.45, m / z found 1011 [M+1]. Step 2. 2-(2-(2-(4-((hexyl(2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (46): [ka]

[0373] To 2-(2-(2-(4-((hexyl(2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate in DCM (5 mL) was added TFA (0.5 mL). The mixture was stirred at room temperature for 2 h. The reaction mixture was diluted with 40 o C was concentrated under reduced pressure and the residue was purified by preparative HPLC (Gemini-C18 150×21.2 mm, 5 um, mobile phase: ACN (0.1% TFA)-HO (0.1% TFA), gradient: 20-70%) to give 2-(2-(2-(4-((hexyl(2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (65.4 mg, 51.8%) as a yellow solid. MS(ESI):mass calcd.for Chemical Formula:C 39 H 56 F6N8O 10 910.40, m / z found 911.4 [M+1]. 1H NMR(400 MHz,MeOD)δ 8.28(s,1H),7.36(m,1H),7.27-7.17(m,1H),5.07(m,1H),4.99(m,1H),4.58(m,4H),4.45(m,2H),4.29(m,4H),4.11-3.94(m,2H),3.9 2-3.87(m,3H),3.77(m,4H),3.64(m,7H),3.27-3.03(m,5H),3.00-2.92(m,1H),2.84(m,1H),1.83(s,2H),1.36(s,6H),0.91(t,J=6.5 Hz,3H). An alternative synthesis of compound (46) is described below.

[0374] Synthesis of 2-(2-(2-(4-((hexyl(2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate tartrate (46 salt) [ka]

[0375] Step 1. 2-(2-(2-azidoethoxy)ethoxy)ethyl (R)-7-(3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate To a solution of 2-(2-(2-azidoethoxy)ethoxy)ethyl (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (Int-22, 300 g, 424 mmol) in ethyl acetate (1 L) was added HCl in ethyl acetate (4 M, 1 L) at room temperature. The mixture was stirred for 2 h and concentrated to give a crude residue. This was redissolved in EA (1.00 L) and quenched with aqueous NaHCO3 to adjust pH = 8, the organic layer was separated, dried over Na2SO4, filtered, and the filtrate was concentrated to give the compound 2-(2-(2-azidoethoxy)ethoxy)ethyl (R)-7-(3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (720 g, 1.12 mol, yield 88.1%, purity 94.5%) as a yellow oil. 1 H NMR:DMSO-d6 7.92(s,1 H),7.52-7.33(m,2 H),5.00-4.95(m,2 H),4.48-4.25(m,10 H),3.94-3.81(m,2 H),3.81-3.78(m,2 H),3.71-3.54(m,14 H),3.39-3.35(m,2 H),2.57-2.56(m,3 H),2.38-2.33(m,4 H),1.40-1.39(m,4 H),1.24-1.17(m,6 H),0.83(t,J=6.8 Hz,3 H). Step 2. 2-(2-(2-(4-((hexyl(2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (46) A solution of compounds 2-(2-(2-azidoethoxy)ethoxy)ethyl (R)-7-(3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (240 g, 395 mmol, 1.00 equiv.) and (2R,3R,4R,5S)-6-(hexyl(prop-2-yn-1-yl)amino)hexane-1,2,3,4,5-pentanol (Int-1, 144 g, 474.63 mmol, 1.20 equiv.) in THF (2.00 L) and HO (1.00 L) was prepared. CuSO4.5H2O (99.0 g, 396 mmol, 1.00 equiv.) and Na ascorbate (78.9 g, 398 mmol, 1.01 equiv.) were added to the mixture at 10-25 °C, and the mixture was stirred at 25 °C for 10 h. The mixture of three batches was diluted with NH3.H2O / NaCl aq. = 1V / 2V (1.50 L), extracted with EA (1.50 L × 2), and washed with NH3.H2O / NaCl aq. = 1V / 2V (1.50 L × 3) until the color of the aqueous phase changed to colorless form blue. The organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to give a residue. The crude product was purified by reverse-phase HPLC (0.1% FA condition), the mixture was concentrated to remove MeCN, then quenched with NaHCO3 to adjust pH = 9, extracted with EA (10.0 L x 2), dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure to give 2-(2-(2-(4-((hexyl(2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (46) (290 g, 306 mmol, 25.9% yield, 96.3% purity) as a yellow solid. MS(ESI):mass calcd.for Chemical Formula:C 39 H 56 F6N8O 10910.40,m / z found 911.5 [M+1].1H NMR(DMSO-d6)7.92(s,1 H),7.52-7.33(m,2 H),5.00-4.95(m,2 H),4.48-4.25(m,10 H),3.94-3.93(m,2 H),3.81-3.79(m,2 H),3.71-3.39(m,14 H),3.35-3.27(m,2 H),2.72-2.54(m,3 H),2.38-2.33(m,4 H),1.55-1.33(m,4 H),1.24-1.15(m,6 H),0.83(t,J=8.0 Hz,3 H). Step 3. 2-(2-(2-(4-((hexyl(2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate tartrate To a solution of 2-(2-(2-(4-((hexyl(2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (260 g, 285 mmol) in ethanol (1 L) was added tartaric acid (43.7 g, 291 mmol, 1.02 equiv.) in ethanol (1 L). The mixture was stirred at 60° C. for 5 h. The mixture was concentrated under reduced pressure to give 2-(2-(2-(4-((hexyl(2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate tartrate as a yellow solid (293.3 g, 93.7% yield, 96.8% purity). 1H NMR:DMSO-d6 8.01-7.95(m,1 H),7.55-7.47(m,2 H),5.02-4.90(m,2 H),4.48(t,J=8.0 Hz,2 H),4.34-3.7(m,9 H),3.81-3.48(m,15 H),3.40-3.35(m,2 H),2.97-2.58(m,5 H),2.46-2.33(m,4 H),1.43-1.42(m,2 H),1.27-1.20(m,6 H),0.83(t,J=6.0 Hz,3 H). [Example 34] Synthesis of 7-((R)-3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-N-(2-(2-(2-(4-((hexyl(2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxamide (47) [ka]

[0376] Step 1: Synthesis of (ethane-1,2-diylbis(oxy))bis(ethane-2,1-diyl)bis(4-methylbenzenesulfonate) [ka]

[0377] To a stirred solution of 2,2'-(ethane-1,2-diylbis(oxy))bis(ethan-1-ol) (15.0 g, 99.88 mmol) in DCM (300 mL) was added EtN (55.58 mL, 399.54 mmol) followed by tosyl chloride (76.16 g, 399.54 mmol) at 0 °C. The reaction mixture was stirred at room temperature for 16 h. After completion of the reaction, the reaction mixture was quenched with cold water (500 mL) and extracted with CHCl (2 x 500 mL). The combined organic layers were washed with cold water (500 mL), brine (500 mL), dried over anhydrous NaSO and concentrated under reduced pressure to give the crude compound. The crude compound was purified by column chromatography on silica gel (100-200 mesh) using 0-50% EtOAc in petroleum ether as eluent to give 37.0 g of (ethane-1,2-diylbis(oxy))bis(ethane-2,1-diyl)bis(4-methylbenzenesulfonate) as an off-white solid. [TLC system: EtOAc:petroleum ether (4:6); Rf value: 0.6].

[0378] Step 2: Synthesis of 1,2-bis(2-azidoethoxy)ethane [ka]

[0379] To a stirred solution of (ethane-1,2-diylbis(oxy))bis(ethane-2,1-diyl)bis(4-methylbenzenesulfonate) (5.0 g, 10.90 mmol) in DMF (50 mL) was added sodium azide (2.83 g, 43.61 mmol). The reaction mixture was then heated at 70° C. for 16 h. After completion of the reaction, the reaction mixture was concentrated and the residue was stirred in diethyl ether and filtered. The filtrate was concentrated to give 2.0 g of 1,2-bis(2-azidoethoxy)ethane as a colorless oil. [TLC system: EtOAc:petroleum ether (2:8); Rf value: 0.6].

[0380] Step 3. 2-(2-(2-azidoethoxy)ethoxy)ethan-1-amine hydrochloride: [ka]

[0381] To a vigorously stirred solution of 1,2-bis(2-azidoethoxy)ethane (3.0 g, 14.98 mmol) in Et2O (22 mL) and THF (4.4 mL) was added a 1N HCl (17.6 mL) solution followed by PPh3 (3.9 g, 14.98 mmol) in Et2O (22 mL). The reaction mixture was stirred at room temperature for 12 h. After completion of the reaction, the aqueous layer was separated and washed with diethyl ether (2 X 100 mL). The aqueous layer was concentrated and the residue was co-evaporated with acetonitrile (2-3 times) to give 2.0 g of 2-(2-(2-azidoethoxy)ethoxy)ethan-1-amine hydrochloride as a colorless oil. [TLC system: MeOH:DCM (0.5:9.5); Rf value: 0.4].

[0382] Step 4. tert-Butyl (R)-(4-(1-((2-(2-(2-azidoethoxy)ethoxy)ethyl)carbamoyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-yl)carbamate [ka]

[0383] To a stirred solution of (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5 trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylic acid (2.9 g, 5.27 mmol) and 2-(2-(2-azidoethoxy)ethoxy)ethan-1-amine hydrochloride (1.66 g, 7.90 mmol) in DMF (30 mL) was added HATU (3.0 g, 7.90 mmol) followed by DIPEA (3.39 mL, 26.35 mmol) at 0° C. The reaction mixture was stirred at room temperature for 45 minutes. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was quenched with cold water (2×50 mL) and extracted with EtOAc (2×100 mL). The combined organic layers were washed with cold water (100 mL), brine (100 mL), dried over anhydrous Na2SO4, and concentrated to give the crude compound. The crude compound was purified by column chromatography on silica gel (Davisil) using 0-70% EtOAc in petroleum ether as eluent to give 3.0 g of tert-butyl (R)-(4-(1-((2-(2-(2-azidoethoxy)ethoxy)ethyl)carbamoyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-yl)carbamate as an off-white solid. [TLC system: EtOAc:petroleum ether (7:3); Rf value: 0.5].

[0384] Step 5 (R)-7-(3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-N-(2-(2-(2-azidoethoxy)ethoxy)ethyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxamide [ka]

[0385] To a solution of tert-butyl (R)-(4-(1-((2-(2-(2-azidoethoxy)ethoxy)ethyl)carbamoyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-yl)carbamate (2.4 g, 3.339 mmol) in CHCl (24 mL) was added 4M HCl in 1,4-dioxane (24.0 mL) at 0° C. The reaction mixture was stirred at room temperature for 6 hours. After completion of the reaction, the reaction mixture was concentrated and the residue was basified with saturated NaHCO3 solution and extracted with 10% MeOH in DCM (2×200 mL). The combined organic layers were dried over anhydrous Na2SO4 and concentrated to give 2.2 g of (R)-7-(3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-N-(2-(2-(2-azidoethoxy)ethoxy)ethyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxamide as an oil. [TLC system: MeOH:DCM (0.5:9.5); R f Value: 0.4].

[0386] Step 6. 7-((R)-3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-N-(2-(2-(2-(4-((hexyl(2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxamide [ka]

[0387] To a solution of (R)-7-(3-amino-4-(2,4,5 trifluorophenyl)butanoyl)-N-(2-(2-(2-azidoethoxy)ethoxy)ethyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxamide (2.2 g, 3.627 mmol) and compound 8 (1.65 g, 5.440 mmol) in t-BuOH (66 mL) and water (22 mL) was added CuSO4, 5.H2O (1.08 g, 4.352 mmol) and Na ascorbate (1.07 g, 5.440 mmol). The reaction mixture was stirred at room temperature for 2 h. After completion of the reaction, the reaction mixture was filtered and concentrated to give the crude compound. The crude compound was purified by RP prep-HPLC to give 0.573 g of 7-((R)-3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-N-(2-(2-(2-(4-((hexyl(2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxamide as an off-white solid.

[0388] RP preparative HPLC purification method: Column: X BRIDGE C18 (19X250mm) Mobile phase A: 10MM ABC in water Mobile phase B: Acetonitrile Solubility: ACN+water+THF Gradient %B: 0 / 10, 1 / 30, 18.5 / 45.4, 18.6 / 95, 20.5 / 95, 20.6 / 10, 22 / 10 LC MS M / Z=910.51(M+1)1H NMR(400 MHz,DMSO-d6)δ 8.02-7.98(m,1H),7.91(s,1H),7.45-7.41(m,2H),4.98-4.87(m,2H),4.47-4.46(m,4H),4.42-4.13(m,4H),3.92(s,2H),3.80(s,2H),3 .72(s,3H),3.62-3.56(m,2H),3.59-3.32(m,12H),2.67-2.56(m,4H),2.40-2.33(m,4H),1.40(bs,2H),1.23-1.15(m,6H),0.83(t,J=7.2 Hz,3H). [Example 35] Synthesis of 2-(2-(2-(4-((bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (48): [ka]

[0389] Compound 2-(2-(2-azidoethoxy)ethoxy)ethyl (R)-7-(3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (0.1 g, 0.164 mmol) and compound (2R,3R, To a solution of 4R,5S)-6-(((2R,3S,4S,5S)-2,3,4,5,6-pentahydroxyhexyl)(prop-2-yn-1-yl)amino)hexane-1,2,3,4,5-pentaol (0.126 g, 0.329 mmol) was added CuSO4, 5.H2O (0.05 g, 0.197 mmol) and Na ascorbate (0.048 g, 0.246 mmol). The reaction was then stirred at room temperature for 16 hours. After completion of the reaction, the reaction mixture was filtered and concentrated to give the crude compound. The crude compound was purified by RP prep HPLC to give 20 mg of 2-(2-(2-(4-((bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-amino-4-(2,4,5 trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate as an off-white solid compound. LC MS M / Z=991.57(M+1) 1 H NMR(400 MHz,DMSO-d6)δ 7.94(s,1H),7.45-7.43(m,2H),5.00-4.95(m,2H),4.87-4.14(m,18H),3.93-3.47(m,23H),3.39-3.16(m,8H),2.61-2.58(m,2H). RP preparative HPLC purification conditions: Column / dimensions: X BRIDGE C18 (19 x 250 mm), 5μ Mobile phase A: 10MM ABC in water (pH) Mobile phase B: Acetonitrile.

[0390] Gradient (% of time / B): 0 / 10, 1 / 10, 11 / 50, 12.6 / 50, 12.7 / 98, 16 / 98, 16.1 / 10, 19 / 10.

[0391] Flow rate: 16ml / min.

[0392] [Example 36] Synthesis of (R)-4-(1-((2-morpholinoethoxy)carbonyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-aminium (2R,3S)-3-carboxy-2,3-dihydroxypropanoate (tartrate) (49): [ka]

[0393] Step 1. 2-Morpholinoethyl (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5 trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate: [ka]

[0394] To a stirred solution of (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylic acid (0.5 g, 0.91 mmol) in 1,2-dichloroethane (10 mL) and triethylamine (0.37 g, 3.64 mmol) was added bis(2-oxo-3-oxazolidinyl)phosphine chloride (0.46 g, 1.82 mmol) at 0° C. and the resulting mixture was stirred at 0° C. for 1 h. Then, 2-morpholinoethan-1-ol (0.24 g, 1.82 mmol) was added at 0° C. and the reaction mixture was stirred at room temperature for 16 h. After completion, the reaction mixture was diluted with water (10 mL) and extracted with dichloromethane (2×50 mL). The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the crude product. The crude residue obtained was then purified by column chromatography on silica gel (230-400 mesh) using 4% methanol in dichloromethane as a gradient to give 2-morpholinoethyl (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (0.51 g, 84.7%) as an off-white solid. TLC system. MeOH:DCM (1:9);R f :0.3. Step 2. 2-Morpholinoethyl (R)-7-(3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate: [ka]

[0395] To a stirred solution of 2-morpholinoethyl (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (0.51 g, 0.77 mmol) in 1,4 dioxane (5.1 mL) was added 4.0 M hydrogen chloride solution in dioxane (5.1 mL) at 0° C. and the reaction mixture was stirred at room temperature for 4 hours. After completion, the reaction mixture was concentrated under reduced pressure to obtain the crude product. The resulting crude product was then purified by RP preparative HPLC using the following conditions: Column / dimensions: X-BRIDGE-C18 (19*250) 5um, Mobile phase A: 10 mM ammonium bicarbonate in water, Mobile phase B: acetonitrile, Gradient (time / % of B): 0 / 5, 1 / 5, 8 / 40, 13.5 / 40, 13.51 / 100, 18 / 100, 18.1 / 5, 20.5 / 5, Flow rate: 18ml / min. The desired fractions were lyophilized to give 2-morpholinoethyl (R)-7-(3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (0.25g, 57.7%) as an off-white semi-solid. TLC system. MeOH:DCM (1:9); R f :0.05. Step 3 (R)-4-(1-((2-morpholinoethoxy)carbonyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-aminium (2R,3S)-3-carboxy-2,3-dihydroxypropanoate-tartrate (49): [ka]

[0396] A mixture of 2-morpholinoethyl (R)-7-(3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (0.25 g, 0.44 mmol) and L(+)-tartaric acid (0.73 g, 0.49 mmol) in demineralized water (2.5 mL) was sonicated to give a clear solution. The resulting solution was lyophilized to give (R)-4-(1-((2-morpholinoethoxy)carbonyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-aminium (2R,3S)-3-carboxy-2,3-dihydroxypropanoate (0.32 g) as an off-white solid. LCMS M / Z 564.46. 1 H NMR(400 MHz,DMSO)δ 7.52(dd,J=9.5,4.0 Hz,2H),5.03-4.87(m,2H),4.35-4.08(m,5H),3.95-3.88(m,4H),3.67(s,2H),3.56(t,J=4.6 Hz,4H),2.89(s,3H),2.82-2.58(m,4H),2.44(t,J=4.7 Hz,4H). [Example 37] Synthesis of (R)-4-(1-((2-(2-morpholinoethoxy)ethyl)carbamoyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-aminium (2R,3R)-3-carboxy-2,3-dihydroxypropanoate (tartrate) (50): [ka]

[0397] Step 1. tert-Butyl (R)-(4-(1-((2-(2-morpholinoethoxy)ethyl)carbamoyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-yl)carbamate: [ka]

[0398] To a stirred solution of (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5 trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylic acid (0.4 g, 0.73 mmol), 2-(2-morpholinoethoxy)ethan-1-amine (0.127 g, 1-[bis(dimethylamino)methylene]-1H-1,2,3 triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (0.334 g, 0.88 mmol) in N,N-dimethylformamide (4 mL) was added N,N-diisopropylethylamine (0.47 g, 3.65 mmol) slowly at 0° C. and the reaction mixture was stirred at room temperature for 4 hours. After completion, the reaction mixture was cooled to room temperature and cooled to room temperature. The mixture was diluted with water (30 mL) and extracted with ethyl acetate (3×100 mL). The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the crude product. The crude residue obtained was then purified by column chromatography on silica gel (230-400 mesh) using 4% methanol in dichloromethane as a gradient to give tert-butyl (R)-(4-(1-((2-(2-morpholinoethoxy)ethyl)carbamoyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-yl)carbamate (0.4 g, 78%) as a brown solid. TLC system. MeOH:DCM (1:9);R f :0.3. Step 2 (R)-7-(3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-N-(2-(2-morpholinoethoxy)ethyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxamide: [ka]

[0399] To a stirred solution of tert-butyl (R)-(4-(1-((2-(2-morpholinoethoxy)ethyl)carbamoyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-yl)carbamate (0.4 g, 0.566 mmol) in 1,4-dioxane (4 mL) was added 4.0 M HCl in dioxane (4 mL) at 0° C. and the mixture was stirred at room temperature for 4 h. After completion, the reaction mixture was concentrated under reduced pressure to give the crude product. The resulting crude product was then purified by RP preparative HPLC using the following conditions: Column / dimensions: Column / dimensions: X-select C18 (19*250mm) 5um, Mobile phase A: 10MM ammonium bicarbonate in water, Mobile phase B: acetonitrile, Gradient (time / % of B): 0 / 10, 1 / 10, 10 / 40, 15.51 / 61.5, 15.6 / 100, 18 / 100, 18.1 / 10, 20 / 10, Flow rate: 17ml / min. The desired fractions were lyophilized to give (R)-7-(3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-N-(2-(2-morpholinoethoxy)ethyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxamide (0.16g, 46.6%) as a colorless gum. TLC system. MeOH:DCM (1:9); f :0.05. Step 3 (R)-4-(1-((2-(2-morpholinoethoxy)ethyl)carbamoyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-aminium (2R,3R)-3-carboxy-2,3-dihydroxypropanoate (tartrate) (50): [ka]

[0400] To a stirred solution of (R)-7-(3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-N-(2-(2-morpholinoethoxy)ethyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxamide (0.16 g, 0.26 mmol) in a mixture of dichloromethane (4 mL) and methanol (1 mL) was added L(+)-tartaric acid (0.039 g, 0.26 mmol) and the mixture was stirred at room temperature for 6 h. The solvent was evaporated under reduced pressure and the residue was then washed with n-pentane (2×5 mL) and redissolved in demineralized water (1.4 mL). The resulting solution was lyophilized to give (R)-4-(1-((2-(2-morpholinoethoxy)ethyl)carbamoyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-aminium (2R,3R)-3-carboxy-2,3-dihydroxypropanoate (0.175 g) as an off-white solid. LCMS M / Z 607.51. 1 H NMR(400 MHz,DMSO)δ 8.08-7.95(m,1H),7.56-7.44(m,2H),5.03-4.87(m,2H),4.26-4.07(m,3H),3.88(s,4H),3.37(t,J=6.1 Hz,13H),2.87(d,J=6.7 Hz,2H),2.72-2.65(m,2H),2.46-2.36(m,7H). [Example 38] Synthesis of (R)-4-(1-((3-morpholinopropoxy)carbonyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-aminium (2R,3R)-3-carboxy-2,3-dihydroxypropanoate (tartrate) (51): [ka]

[0401] Step 1. 3-Morpholinopropyl (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate: [ka]

[0402] To a stirred solution of (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylic acid (0.3 g, 0.545 mmol) in 1,2-dichloroethane (6 mL) and triethylamine (0.165 g, 1.63 mmol) was added bis(2-oxo-3-oxazolidinyl)phosphine chloride (0.207 g, 0.817 mmol) at 0° C. and the resulting mixture was stirred at 0° C. for 1 h. Then, 3-morpholinopropan-1-ol (0.119 g, 0.817 mmol) was added at 0° C. and the reaction mixture was stirred at room temperature for 16 h. After completion, the reaction mixture was diluted with water (10 mL) and extracted with dichloromethane (2×30 mL). The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the crude product. The crude residue obtained was then purified by column chromatography on silica gel (230-400 mesh) using 4% methanol in dichloromethane as a gradient to give 3-morpholinopropyl (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (0.21 g, 57.2%) as an off-white solid. TLC system. MeOH:DCM (1:9);R f :0.3. Step 2. 3-Morpholinopropyl (R)-7-(3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate: [ka]

[0403] To a stirred solution of 3-morpholinopropyl (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (0.21 g, 0.31 mmol) in 1,4 dioxane (2 mL) was added 4.0 M in dioxane (2 mL) at 0° C. and the reaction mixture was stirred at room temperature for 4 hours. After completion, the reaction mixture was concentrated under reduced pressure to obtain the crude product. The obtained crude product was purified by RP preparative HPLC using the following conditions: Column / dimensions: X select C18 (19*250mm) 5u, Mobile phase A: 10mM ammonium bicarbonate in water, Mobile phase B: acetonitrile, Gradient (time / % of B): 0.01 / 25, 1 / 25, 9 / 40, 13 / 40, 13.1 / 100, 18 / 100, 18.1 / 25, 20 / 25, Flow rate: 18ml / min. The desired fractions were lyophilized to give 3-morpholinopropyl(R)-7-(3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (0.125g, 69.8%) as a colorless gum. TLC system. MeOH:DCM (1:9);R f :0.05. Step 3 (R)-4-(1-((3-morpholinopropoxy)carbonyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-aminium (2R,3R)-3-carboxy-2,3-dihydroxypropanoate (51): [ka]

[0404] To a stirred solution of 3-morpholinopropyl (R)-7-(3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (0.125 g, 0.216 mmol) in a mixture of dichloromethane (4 mL) and methanol (1.0 mL) was added L(+)-tartaric acid (0.032 g, 0.216 mmol) and the mixture was stirred at room temperature for 6 h. The reaction mixture was concentrated and washed with n-pentane (2×5 mL). The resulting residue was dissolved in water and lyophilized to give (R)-4-(1-((3-morpholinopropoxy)carbonyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-aminium (2R,3R)-3-carboxy-2,3-dihydroxypropanoate (0.077 g) as an off-white solid. LCMS M / Z 578.22. 1 H NMR(400 MHz,DMSO)δ 7.56-7.46(m,2H),4.94(d,J=11.9 Hz,2H),4.28(t,J=6.6 Hz,5H),3.90(s,6H),3.56(s,4H),2.86(s,3H),2.72-2.66(m,2H),2.38(dd,J=13.7,6.5 Hz,6H),1.89-1.80(m,2H). [Example 39] Synthesis of (R)-4-(1-((2-(2-morpholinoethoxy)ethoxy)carbonyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-aminium (2R,3R)-3-carboxy-2,3-dihydroxypropanoate (tartrate) (52): [ka]

[0405] Step 1. 2-(2-morpholinoethoxy)ethyl (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5 trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate: [ka]

[0406] To a stirred solution of (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine, a mixture of 1-carboxylic acid (0.3 g, 0.54 mmol) in 1,2-dichloroethane (6 mL) and triethylamine (0.27 g, 2.7 mmol) was added bis(2-oxo-3-oxazolidinyl)phosphine chloride (0.28 g, 1.09 mmol) at 0° C. and the resulting mixture was stirred at 0° C. for 1 h. Then, 2-(2-morpholinoethoxy)ethan-1-ol (0.191 g, 1.09 mmol) was added at 0° C. and the reaction mixture was stirred at room temperature for 16 h. After completion, the reaction mixture was diluted with water (10 mL) and extracted with dichloromethane (2×30 mL). The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the crude product. The crude residue obtained was then purified by column chromatography on silica gel (230-400 mesh) using 4% methanol in dichloromethane as a gradient to give 2 2-(2-morpholinoethoxy)ethyl (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (0.32 g, 83.1%) as an off-white solid. TLC system. MeOH:DCM (1:9);R f :0.3. Step 2. 2-(2-morpholinoethoxy)ethyl (R)-7-(3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate: [ka]

[0407] To a stirred solution of 2-(2-morpholiethoxy)ethyl (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (0.32 g, 0.45 mmol) in 1,4 dioxane (3.2 mL) was added 4.0 M HCl in dioxane (3.2 mL) at 0° C. and the mixture was stirred at room temperature for 4 hours. After completion, the reaction mixture was concentrated under reduced pressure to give the crude product. The crude product was then purified by reverse phase preparative HPLC using the following conditions: Column / dimensions: X-bridge-C18 (19*250mm) 5um, Mobile phase A: 10mM ammonium bicarbonate in water, Mobile phase B: acetonitrile, Gradient (time / % of B): 0 / 5, 1 / 5, 12 / 75, 14 / 75, 14.05 / 98, 16.50 / 98, 16.51 / 5, 19 / 5, Flow rate: 18ml / min. The desired fractions were lyophilized to give 2-(2-morpholinoethoxy)ethyl (R)-7-(3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (0.14g, 51.1%) as a colorless gum. TLC system. MeOH:DCM (1:9);R f :0.05. Step 3 (R)-4-(1-((2-(2-morpholinoethoxy)ethoxy)carbonyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-aminium (2R,3R)-3-carboxy-2,3-dihydroxypropanoate (tartrate) (52): [ka]

[0408] A mixture of 2-(2-morpholinoethoxy)ethyl (R)-7-(3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (0.14 g, 0.23 mmol) and L(+)-tartaric acid (0.038 g, 0.25 mmol) in demineralized water (1.4 mL) was sonicated to give a clear solution. The resulting solution was lyophilized to give (R)-4-(1-((2-(2-morpholinoethoxy)ethoxy)carbonyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-aminium (2R,3R)-3-carboxy-2,3-dihydroxypropanoate (0.172 g) as an off-white solid. LCMS M / Z 608.46. 1 H NMR(400 MHz,DMSO)δ 7.53(q,J=9.6 Hz,2H),4.95(q,J=18.0 Hz,2H),4.40-4.05(m,6H),3.92(d,J=12.6 Hz,5H),3.68(t,J=4.8 Hz,5H),3.50(t,J=4.6 Hz,4H),2.73(s,2H),2.46(d,J=5.7 Hz,3H),2.38(s,4H). [Example 40] Synthesis of (R)-4-(1-(((5-morpholinopentyl)oxy)carbonyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-aminium (2R,3R)-3-carboxy-2,3-dihydroxypropanoate (tartrate) (53): [ka]

[0409] Step 1. 5-Morpholinopentyl (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate: [ka]

[0410] To a stirred solution of (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylic acid (1.0 g, 1.82 mmol) in 1,2-dichloroethane (20 mL) and triethylamine (1.27 g, 9.1 mmol) was added bis(2-oxo-3-oxazolidinyl)phosphine chloride (0.93 g, 3.64 mmol) at 0° C., and the resulting mixture was stirred at 0° C. for 1 h. Then, 5-morpholinopentan-1-ol (0.94 g, 5.45 mmol) was added at 0° C., and the mixture was stirred at room temperature for 16 h. After completion, the reaction mixture was diluted with water (10 mL) and extracted with dichloromethane (2×30 mL). The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the crude product. The crude residue obtained was then purified by column chromatography on silica gel (230-400 mesh) using 4% methanol in dichloromethane as a gradient to give 5-morpholinopentyl (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (0.6 g, 46.9%) as a colorless gum. TLC system. MeOH:DCM (1:9);R f :0.3. Step 2. 5-Morpholinopentyl (R)-7-(3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate: [ka]

[0411] To a stirred solution of 5-morpholinopentyl (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (0.6 g, 0.85 mmol) in 1,4 dioxane (6 mL) was added 4.0 M HCl in dioxane (6 mL) at 0° C. and the reaction mixture was stirred at room temperature for 4 h. After completion, the reaction mixture was concentrated under reduced pressure to give the crude product. The crude product was then purified by RP preparative HPLC using the following conditions: Column / dimensions: X select C18 (19*250mm) 5um, Mobile phase A: 10mM ammonium bicarbonate in water, Mobile phase B: acetonitrile, Gradient (time / % of B): 0 / 20, 1 / 20, 12 / 50, 16.5 / 50, 16.6 / 100, 19 / 100, 19.1 / 20, 21 / 20, Flow rate: 18ml / min. The desired fractions were lyophilized to give 5-morpholinopentyl(R)-7-(3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (0.29g, 56.3%) as a colorless gum. TLC system. MeOH:DCM (1:9);R f :0.05. Step 3 (R)-4-(1-(((5-morpholinopentyl)oxy)carbonyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-aminium (2R,3R)-3-carboxy-2,3-dihydroxypropanoate (tartrate) (53): [ka]

[0412] A mixture of 5-morpholinopentyl (R)-7-(3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (0.19 g, 0.31 mmol) and L(+)-tartaric acid (0.052 g, 0.34 mmol) in demineralized water (1.9 mL) was sonicated for 2 min to give a clear solution. The resulting solution was lyophilized to give (R)-4-(1-(((5-morpholinopentyl)oxy)carbonyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-aminium (2R,3R)-3-carboxy-2,3-dihydroxypropanoate (0.23 g) as an off-white solid. LCMS M / Z 606.48. 1 H NMR(400 MHz,DMSO)δ 7.51(td,J=10.2,6.9 Hz,2H),5.00-4.88(m,2H),4.24(td,J=7.2,3.4 Hz,5H),3.99-3.87(m,5H),3.68(s,2H),3.54(t,J=4.5 Hz,5H),2.90(s,2H),2.80-2.64(m,2H),2.38-2.26(m,6H),1.68(p,J=7.3 Hz,2H),1.55-1.34(m,4H). [Example 41] Synthesis of (R)-4-(1-((4-morpholinobutoxy)carbonyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-aminium (2R,3R)-3-carboxy-2,3-dihydroxypropanoate (tartrate) (54): [ka]

[0413] Step 1. 4-Morpholinobutyl (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5 trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate: [ka]

[0414] To a stirred solution of (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylic acid (0.5 g, 0.91 mmol) in 1,2-dichloroethane (10 mL) and triethylamine (0.37 g, 3.64 mmol) was added bis(2-oxo-3-oxazolidinyl)phosphine chloride (0.35 g, 1.36 mmol) at 0° C. and the mixture was stirred at 0° C. for 1 h. Then, 4-morpholinobutan-1-ol (0.22 g, 1.36 mmol) was added at 0° C. and the reaction mixture was stirred at room temperature for 16 h. After completion, the reaction mixture was diluted with water (10 mL) and extracted with dichloromethane (2×30 mL). The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the crude product. The crude residue was then purified by column chromatography on silica gel (230-400 mesh) using 4% methanol in dichloromethane as eluent to give 4-morpholinobutyl (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (0.45 g, 71.6%) as an off-white solid. TLC system. MeOH:DCM (1:9);R f :0.3. Step 2. 4-Morpholinobutyl (R)-7-(3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate: [ka]

[0415] To a stirred solution of 4-morpholinobutyl (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (0.45 g, 0.65 mmol) in 1,4 dioxane (4.5 mL) was added 4.0 M HCl in dioxane (4.5 mL) at 0° C. and the reaction mixture was stirred at room temperature for 4 h. After completion, the reaction mixture was concentrated under reduced pressure to give the crude product. The crude product was then purified by RP preparative HPLC using the following conditions: Column / dimensions: X-bridge C18 (19*250mm) 5μ, Mobile phase A: 10MM ammonium bicarbonate in water, Mobile phase B: Acetonitrile (org), Gradient (time / % of B): 0 / 20, 2 / 20, 10 / 45, 14 / 45, 14.10 / 100, 16 / 100, 16.10 / 20, 18 / 20, Flow rate: 17ml / min. The desired fractions were lyophilized to give 4-morpholinobutyl (R)-7-(3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (0.16g, 41.7%) as a colorless gum. TLC system. MeOH:DCM (1:9); R f :0.05. Step 3 (R)-4-(1-((4-morpholinobutoxy)carbonyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-aminium (2R,3R)-3-carboxy-2,3-dihydroxypropanoate (tartrate) (54): [ka]

[0416] A mixture of 5-morpholinopentyl 4-morpholinobutyl (R)-7-(3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (0.09 g, 0.15 mmol) and L(+)-tartaric acid (0.025 g, 0.167 mmol) in demineralized water (1 mL) was sonicated to give a clear solution. The resulting solution was lyophilized to give (R)-4-(1-((4-morpholinobutoxy)carbonyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-aminium (2R,3R)-3-carboxy-2,3-dihydroxypropanoate (0.115 g) as an off-white solid. LCMS M / Z 592.49. 1 H NMR(400 MHz,DMSO)δ 7.53(q,J=8.2 Hz,2H),4.94(d,J=11.9 Hz,2H),4.25(dd,J=7.7,5.3 Hz,5H),3.93(s,5H),3.64(d,J=6.5 Hz,5H),2.87(d,J=7.4 Hz,2H),2.78-2.64(m,2H),2.45-2.13(m,6H),1.68(d,J=8.2 Hz,2H),1.54(q,J=7.4 Hz,2H). [Example 42] Synthesis of (R)-4-(1-((2-methyl-2-azaspiro[3.3]heptan-6-yl)carbamoyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-aminium (2R,3R)-3-carboxy-2,3-dihydroxypropanoate (tartrate) (55): [ka]

[0417] Step 1 Tert-butyl (R)-(4-(1-((2-methyl-2-azaspiro[3.3]heptan-6-yl)carbamoyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-yl)carbamate: [ka]

[0418] (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylic acid (0.5 g, 0.91 mmol), 2-methyl-2-azaspiro[3.3]heptan-6-amine salt in N,N-dimethylformamide (5 mL). To a stirred solution of 1-[bis(dimethylamino)methylene]-1H-1,2,3 triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (0.415 g, 0.1.09 mmol) and 1,2,3-bis(dimethylamino)methylene-1H-1,2,3 triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (0.415 g, 0.1.09 mmol) at 0° C., N,N-diisopropylethylamine (0.59 g, 4.55 mmol) was slowly added and the reaction mixture was stirred at room temperature for 4 hours. After completion of the reaction, The mixture was diluted with water (30 mL) and extracted with ethyl acetate (3×100 mL). The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the crude product. The crude residue obtained was then purified by column chromatography on silica gel (230-400 mesh) using 4% methanol in dichloromethane as eluent to give tert-butyl (R)-(4-(1-((2-methyl-2-azaspiro[3.3]heptan-6-yl)carbamoyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-yl)carbamate (0.3 g, 50.2%) as an off-white solid. TLC system. MeOH:DCM (1:9);Rf :0.3. Step 2 (R)-7-(3-amino)-4-(2,4,5-trifluorophenyl)butanoyl)-N-(2-methyl-2-azabicyclo[3.3]heptan-6-yl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxamide: [ka]

[0419] To a stirred solution of tert-butyl (R)-(4-(1-((2-methyl-2-azaspiro[3.3]heptan-6-yl)carbamoyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-yl)carbamate (0.3 g, 0.45 mmol) in 1,4-dioxane (3 mL) was added 4.0 M HCl in dioxane (3 mL) at 0° C. and the mixture was stirred at room temperature for 4 h. After completion, the reaction mixture was concentrated under reduced pressure to give the crude product. The resulting crude product was then purified by RP preparative HPLC using the following conditions: Column / dimensions: X-select C18 (19*250mm) 5μ, Mobile phase A: 10MM ammonium bicarbonate in water, Mobile phase B: acetonitrile (org), Gradient (time / % of B): 0 / 10, 1 / 10, 10 / 40, 16 / 63.2, 16.1 / 100, 19 / 100, 19.1 / 10, 22 / 10, Flow rate: 17ml / min. The desired fractions were lyophilized to give (R)-7-(3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-N-(2-methyl-2-azaspiro[3.3]heptan-6-yl)3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxamide (0.09g, 35.4%) as an off-white solid. TLC system. MeOH:DCM (1:9); f :0.05. Step 3 (R)-4-(1-((2-methyl-2-azaspiro[3.3]heptan-6-yl)carbamoyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-aminium (2R,3R)-3-carboxy-2,3-dihydroxypropanoate (tartrate) (55) [ka]

[0420] A mixture of (R)-7-(3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-N-(2-methyl-2-azaspiro[3.3]heptan-6-yl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxamide (0.14 g, 0.25 mmol) and methanol (1 mL) mixture L(+) tartaric acid (0.038 g, 0.25 mmol) in dichloromethane (4 mL) was added and the reaction mixture was stirred at room temperature for 6 hours. The reaction mixture was concentrated and the residue was washed with n-pentane (2 X 5 mL) and the residue was dissolved in demineralized water (3 mL). The resulting solution was lyophilized to give the synthesis of (R)-4-(1-((2-methyl-2-azaspiro[3.3]heptan-6-yl)carbamoyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazin-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-aminium (2R,3R)-3-carboxy-2,3-dihydroxypropanoate (0.155 g) as an off-white solid. LCMS M / Z 559.46. 1 H NMR(400 MHz,DMSO)δ 8.40-8.22(m,1H),7.48(q,J=9.2 Hz,2H),4.91(d,J=14.9 Hz,2H),4.26(dd,J=16.4,8.0 Hz,3H),4.11(s,1H),3.89(s,3H),3.83(s,3H),2.78(d,J=7.1 Hz,2H),2.70-2.56(m,3H),2.45-2.28(m,7H). [Example 45] In vitro and additional characterization data DPP4 activity assay. Human DPP4 activity assay data was obtained using a DPP4 activity assay kit (Sigma-Aldrich, MAK088) according to the manufacturer's instructions. Briefly, 10 μL of DPP4 assay buffer was transferred per well into a low-volume 384-well plate before transferring 10 μL of test compound dissolved in DPP4 assay buffer. 5 μL of master reaction mix was added to each well, which contains a fluorescent substrate that becomes fluorescent upon enzymatic cleavage. Fluorescence intensity measurements were recorded at 1-minute intervals over the course of 20 minutes using an Envision multimode plate reader (PerkinElmer). The results are shown in Table 4 below.

[0421] AEC2 proliferation assay. Primary human AEC2 were plated at a density of 1,500 cells per well in black 384-well plates (Greiner) coated with 10ug / mL laminin (Life Technologies) in 50μL of small airway epithelial cell growth medium (Lonza) without EGF, retinoic acid, and with 5% BPE. 100nL of test compound dissolved in DMSO was then delivered using a Biomek FX instrument (Beckman Coulter) fitted with a pin tool head (V&P Scientific). After 96 hours of growth at 37°C, cells were fixed with 4% paraformaldehyde, washed three times with PBS, and then immunostained for KI-67 positivity (1:1000, Abcam, ab15580) overnight at 4°C. After three further washes, cells were incubated with a secondary AlexaFluor-conjugated secondary antibody for 1 h at room temperature and then exposed to 10 μg / mL Hoechst 33342 (Life Technologies). Plates were sealed and then quantitative high-content imaging was performed on a CellInsight CX5 HCS instrument (ThermoFisher). The ACE2 proliferation concentration curve for compound 46 is shown in Figure 1. [Table 6] TIFF2024504328000233.tif201153TIFF2024504328000234.tif190154TIFF2024504328000235.tif22 7154TIFF2024504328000236.tif133154TIFF2024504328000237.tif184154TIFF2024504328000238.t if161153TIFF2024504328000239.tif235154TIFF2024504328000240.tif149153TIFF20245043280002 41.tif199153TIFF2024504328000242.tif202153TIFF2024504328000243.tif215154TIFF20245043280 00244.tif198153TIFF2024504328000245.tif167153TIFF2024504328000246.tif209153TIFF2024504 328000247.tif163154TIFF2024504328000248.tif167153TIFF2024504328000249.tif190154TIFF202 4504328000250.tif202154TIFF2024504328000251.tif200154TIFF2024504328000252.tif200154TIF F2024504328000253.tif188154TIFF2024504328000254.tif218154TIFF2024504328000255.tif158154

[0422] [Example 46] Pharmacokinetic profiling To evaluate the time course of plasma and lung exposure of the compounds disclosed herein, rodents were IT-administered with exemplary compounds of the present disclosure. Plasma and lung samples were collected at different time points. Drug levels were measured by LCMS.

[0423] When administered IT to mice, compound 46 demonstrated significantly higher plasma and lung exposure profiles compared to retagliptin (Figure 2). Furthermore, the compound was retained in the lungs for 7 days, whereas pulmonary retagliptin levels were very low after 48 hours.

[0424] Table 5 shows the pharmacokinetic parameters of compound 46 in mice and rats. [Table 7]

[0425] [Example 47] In vivo efficacy studies Experimental method for the ALI model LPS from Escherichia coli I111:B4 (Sigma) was used to induce acute lung injury in mice. Weight-matched (19 grams to 22 grams) 9-11 week-old female C57BL / 6J mice were selected for use in the ALI model.

[0426] For oral delivery, DPP4 inhibitors were dissolved in PBS to obtain a clear solution. Vehicle control or DPP4 inhibitors were administered at 10 ml / kg by oral gavage once or twice daily, selected based on PK profile. For intratracheal delivery of DPP4 inhibitors, compounds were dissolved in PBS to obtain a clear solution. Vehicle control or DPP4 inhibitors were administered every other day at 2 ml / kg through a 22 g flexible catheter.

[0427] LPS (1.5 mg / kg to test orally delivered DPP4 inhibitors and 1.2 mg / kg to test intratracheally delivered DPP4 inhibitors) or PBS in the sham group was injected intratracheally into the lungs of mice on day 0. DPP4 inhibitors or vehicle control were administered to mice starting 1 day before LPS injection (day -1).

[0428] All animals were sacrificed 3.5 days after LPS injection. Bronchoalveolar lavage fluid (BALF) was collected using standard methods. Lungs were inflated with 1 ml of 4% formalin, then fixed in 4% formalin for 24 hours and stored in 70% EtOH until histological processing.

[0429] For readout, the total protein content in BALF was quantified using the BCA assay. Lung inflammation and injury were assessed using H&E staining.

[0430] Experimental method of the bleomycin model Bleomycin (Hospira) was used to induce pulmonary fibrosis in mice. Weight-matched (24 grams to 28 grams) 10-12 week old male C57BL / 6J mice were selected for use in the bleomycin model.

[0431] For oral delivery, DPP4 inhibitors were dissolved in PBS to obtain a clear solution. Vehicle control or DPP4 inhibitors were administered at 10 ml / kg by oral gavage once or twice daily, selected based on PK profile. For intratracheal delivery of DPP4 inhibitors, compounds were dissolved in PBS to obtain a clear solution. Vehicle control or DPP4 inhibitors were administered every 4 days at 2 ml / kg through a 22 g flexible catheter.

[0432] 0.5 U / kg bleomycin or PBS in the sham group was injected intratracheally into the mouse lungs on day 0. DPP4 inhibitors or vehicle control were administered to the mice starting one day before bleomycin injection (day -1).

[0433] All animals were sacrificed 20 days after bleomycin injection. Bronchoalveolar lavage fluid (BALF) was collected using standard methods. Lungs were inflated with 1 ml of 4% formalin, then fixed in 4% formalin for 24 hours and stored in 70% EtOH until histological processing.

[0434] For readings, body weight was measured daily. Total protein content in BALF was quantified using BCA assay. Pulmonary fibrosis was evaluated using Masson's trichrome staining.

[0435] In the mouse ALI model, compound 46 demonstrated a minimally effective dose of 0.02 mg / kg administered intratracheally every 2 days. Compound 46 was also effective in a bleomycin-induced pulmonary fibrosis model, showing rescue of key metrics including body weight, BALF protein content, fibrotic area and histological scoring (Figure 3). The minimally effective dose per 4-day intratracheal administration regimen was 0.5 mg / kg.

[0436] Further evaluation of the efficacy of compound 46 in combination with the standard of care drug nintedanib in a rodent bleomycin model was determined (Figure 4). The combination of compound 46 with nintedanib demonstrated impressive and synergistic efficacy as determined by Bliss independence calculations on metrics of BALF protein content and Ashcroft score of fibrosis severity.

[0437] To understand which cell types were proliferating in response to compound 46 treatment, we performed single-cell RNA sequencing from mouse lungs. After treatment with the compound (0.5 mg / kg IT, animals were sacrificed 2 and 4 days after administration), only circulating and transitional AEC2s were induced to proliferate among other cell populations, including other epithelial cell types but excluding actively expanding immune cells (Figure 5A, Figure 5B). The data further demonstrate a time-dependent accumulation of AEC2s after treatment with the compound.

Claims

1. A compound of formula (I): During the ceremony each --, when optionally present, represents a single bond forming a fused cyclopropyl ring; L １Ａ But -NHCH ２ - or -CH(NH ２ )—and X １ is -O-, -S-, -S(O)-, S(O) ２ - and -NH-; L １Ｂ is one or more -CH ２ - is optionally independently substituted by a moiety selected from -O-, -C(O)-, and -NH-; ２ -C １２ - alkyl; Z １ But, H, C ６ -C １０ -aryl, and 5-10 membered heteroaryl, where 1-4 heteroaryl members are independently selected from N, O and S; m1 is Z １ is 0 if H, and Z １ is an integer which is 1 if other than H; n1 is an integer selected from 0, 1, 2, and 3; R １ But, H, C １ -C １０ -alkyl, and -C １ -C １０ -Alkyl-(C ６ -C １０ -aryl), optionally substituted with 1 to 6 -OH; R ２ is C substituted with 1 to 6 -OH １ -C １０ -alkyl; or A compound of formula (II): During the ceremony W is CH or N; o is an integer selected from 1, 2, and 3; R ３ But, C １ -C ６ -Alkyl, C １ -C ６ -hydroxyalkyl, and -(CH ２ CH ２ O) ｘ H, where x is an integer selected from 1, 2, 3, 4, and 5; R ４ But, C ２ -C ８ -alkynyl; R ５ａ , R ５ｂ , R ５ｃ , and R ５ｄ But, H, C １ -C ６ -Alkyl, halo, -NR Ａ R Ｂ (R Ａ and R Ｂ is H and C １ -C １０ -alkyl), -C(O)OH, -B(OH) ２ , -C(O)NR Ａ R Ｂ , -C(O)OR Ａ , and L ２ C ２ -C １２ -alkyl, with one or more -CH ２ - is optionally independently substituted by a moiety selected from -O-, -C(O)-, and -NH-; ２ -Z ２ - [(CH ２ ) ｎ２ -NR ６ R ７ ] ｍ２ are independently selected from; Z ２ But, H, C ６ -C １０ -aryl, and 5-10 membered heteroaryl, where 1-4 heteroaryl members are independently selected from N, O, and S; R ６ But, H, C １ -C １０ -alkyl, and -C １ -C １０ -Alkyl-(C ６ -C １０ -aryl), optionally substituted with 1 to 6 -OH; R ７ is C substituted with 1 to 6 -OH １ -C １０ - alkyl; m2 is Z １ is 0 if H, and Z １ is an integer which is 1 if other than H; n2 is an integer selected from 0, 1, 2, and 3; R ５ａ , R ５ｂ , R ５ｃ , and R ５ｄ is other than H; If W is CH, then R ５ａ and R ５ｄ is not selected from -C(O)OH, -C(O)OMe, and -C(O)OEt; or A compound of formula (III): During the ceremony X ３ is -O- or -NH-; L ３ is a bond or one or more -CH ２ - is optionally independently substituted by a moiety selected from -O-, -C(O)-, and -NH-; ２ -C １２ - alkyl; Z ３ H, -N ３ , , C ６ -C １０ - selected from aryl, 5-10 membered heteroaryl, where 1-4 heteroaryl members are independently selected from N, O, and S, and (3-14 membered heterocycloalkyl, where 1-4 ring members are independently selected from N, O, and S; Heteroaryl and heterocycloalkyl are selected from halo, NO ２ , OH, CN, and C １ -C ６ - optionally substituted with 1 to 6 substituents selected from the group consisting of haloalkyl; m3 is Z ３ is H or -N ３ 0 when Z ３ is H or -N ３ is an integer that is 1 otherwise; n3 is an integer selected from 0, 1, 2, and 3; R ８ But, H, C １ -C １０ -alkyl, and -C １ -C １０ -Alkyl-(C ６ -C １０ -aryl), optionally substituted with 1 to 6 -OH; R ９ is C substituted with 1 to 6 -OH １ -C １０ - alkyl; R １０ But, C １ -C ６ -haloalkyl; Each R １１ But, H, C １ -C ６ - independently selected from alkyl, alkyl, and halo; o3 is an integer selected from 0, 1, 2, and 3; p3 is an integer selected from 0, 1, 2, and 3; A compound wherein q3 is an integer selected from 0, 1, 2, and 3; or a pharma- ceutically acceptable salt thereof, However, the following compounds are excluded: 【Table 1】

2. 2. The compound of claim 1, which is of formula (I) or a pharma- ceutically acceptable form thereof.

3. L １Ａ Ga-NHCH ２ 3. The compound of claim 2, which is: or a pharma- ceutically acceptable version thereof.

4. L １Ａ -CH(NH ２ 3. The compound of claim 2, wherein:

5. X １ The compound according to any one of claims 2 to 4, or a pharma- ceutically acceptable form thereof, wherein

6. Z １ The compound according to any one of claims 2 to 5, wherein is H, or a pharma- ceutically acceptable form thereof.

7. Z １ C ６ -C １０ -aryl or 5-10 membered heteroaryl, in which 1-4 heteroaryl members are independently selected from N, O and S, or a pharma- ceutically acceptable version thereof.

8. Z １ The compound according to any one of claims 2 to 5 and 7, or a pharma- ceutically acceptable form thereof, wherein is phenyl.

9. Z １ The compound according to any one of claims 2 to 5 and 7, or a pharma- ceutically acceptable form thereof, wherein is triazolyl.

10. 10. The compound according to any one of claims 7 to 9, wherein n1 is 1 or 2, or a pharma- ceutically acceptable form thereof.

11. R １ C optionally substituted with 1 to 6 —OH １ -C １０ 11. The compound according to any one of claims 7 to 10, or a pharma- ceutically acceptable form thereof, wherein:

12. R １ C １ -C ６ 12. The compound according to any one of claims 7 to 11, or a pharma- ceutically acceptable form thereof, wherein: - alkyl.

13. R ２ is substituted with 1 to 5 -OH ２ -C ６ 13. The compound according to any one of claims 7 to 12, or a pharma- ceutically acceptable form thereof, wherein: - alkyl.

14. R ２ is substituted with 3 to 5 -OH ２ -C ６ 14. The compound according to any one of claims 7 to 13, or a pharma- ceutically acceptable form thereof, wherein: - alkyl.

15. R ２ but:

15. The compound according to any one of claims 7 to 14, which is:

16. Z １ But, C ６ -C １０ -aryl or 5-10 membered heteroaryl, where 1-4 heteroaryl members are independently selected from N, O and S; n1 is 1 or 2; R １ But, C １ -C １０ - alkyl; R ２ is C substituted with 1 to 5 -OH ２ -C ６ -alkyl.

17. 3. A compound according to claim 1 or 2, which is selected from the table below, or a pharma- ceutically acceptable version thereof.

18. 2. The compound of claim 1, which is of formula (II) or a pharma- ceutically acceptable form thereof.

19. 20. The compound of claim 18, or a pharma- ceutically acceptable form thereof, wherein W is CH.

20. 20. The compound of claim 18, or a pharma- ceutically acceptable form thereof, wherein W is N.

21. R ５ｂ and R ５ｄ The compound according to any one of claims 18 to 20, or a pharma- ceutically acceptable form thereof, wherein

22. R ５ａ is -C(O)OH or -C(O)OR Ａ 22. The compound according to any one of claims 18 to 21, which is:

23. R ５ａ Ga-C(O)-L ２ -Z ２ - [(CH ２ ) ｎ２ -NR ６ R ７ ] ｍ２ 22. The compound according to any one of claims 18 to 21, which is:

24. Z ２ C ６ -C １０ -aryl or 5-10 membered heteroaryl, in which 1-4 heteroaryl members are independently selected from N, O and S; or a pharma- ceutically acceptable version thereof.

25. Z ２ 25. The compound according to any one of claims 18 to 24, or a pharma- ceutically acceptable form thereof, wherein is phenyl or triazolyl.

26. Z ２ 26. The compound according to any one of claims 18 to 25, or a pharma- ceutically acceptable form thereof, wherein is triazolyl.

27. 27. The compound according to any one of claims 18 to 26, or a pharma- ceutically acceptable form thereof, wherein n2 is 1 or 2.

28. R ６ C optionally substituted with 1 to 6 —OH １ -C １０ 28. The compound according to any one of claims 18 to 27, or a pharma- ceutically acceptable form thereof, wherein:

29. R ６ C １ -C ６ 29. The compound according to any one of claims 18 to 28, or a pharma- ceutically acceptable form thereof, wherein: - alkyl.

30. R ７ is substituted with 1 to 5 -OH ２ -C ６ 30. The compound according to any one of claims 18 to 29, or a pharma- ceutically acceptable form thereof, wherein: - alkyl.

31. R ７ is substituted with 3 to 5 -OH ２ -C ６ 31. The compound according to any one of claims 18 to 30, or a pharma- ceutically acceptable form thereof, wherein: - alkyl.

32. R ７ but:

32. The compound according to any one of claims 18 to 31, which is:

33. R ５ａ , R ５ｂ , R ５ｃ and R ５ｄ One of them is -C(O)-L ２ -Z ２ - [(CH ２ ) ｎ２ -NR ６ R ７ ] ｍ２ and Z ２ But, C ６ -C １０ -aryl or 5-10 membered heteroaryl, where 1-4 heteroaryl members are independently selected from N, O and S; n2 is 1 or 2; R ６ But, C １ -C １０ - alkyl; R ７ is C substituted with 1 to 5 -OH ２ -C ６ - alkyl.

34. 20. A compound according to claim 1 or 18, which is selected from the table below, or a pharma- ceutically acceptable version thereof.

35. 2. The compound of claim 1, which is of formula (III) or a pharma- ceutically acceptable form thereof.

36. X ３ is O, or a pharma- ceutically acceptable form thereof.

37. X ３ or a pharma- ceutically acceptable form thereof.

38. Z ３ Ga-N ３ 38. The compound according to any one of claims 35 to 37, which is:

39. Z ３ C ６ -C １０ -aryl or 5-10 membered heteroaryl, in which 1-4 heteroaryl members are independently selected from N, O and S; or a pharma- ceutically acceptable version thereof.

40. 40. The compound of any one of claims 35 to 39, or a pharma- ceutically acceptable form thereof, wherein each of p3 and q3 is 1.

41. Z ３ 41. The compound according to any one of claims 35 to 40, or a pharma- ceutically acceptable form thereof, wherein is phenyl or triazolyl.

42. Z ３ 42. The compound of any one of claims 35 to 41, or a pharma- ceutically acceptable form thereof, wherein is triazolyl.

43. 43. The compound according to any one of claims 35 to 42, or a pharma- ceutically acceptable form thereof, wherein n3 is 1 or 2.

44. R ８ C optionally substituted with 1 to 6 —OH １ -C １０ 44. The compound according to any one of claims 35 to 43, or a pharma- ceutically acceptable form thereof, wherein: - alkyl.

45. R ８ C １ -C ６ 45. The compound according to any one of claims 35 to 44, or a pharma- ceutically acceptable form thereof, wherein: - alkyl.

46. R ９ is substituted with 1 to 5 -OH ２ -C ６ 46. ​​The compound of any one of claims 35 to 45, or a pharma- ceutically acceptable form thereof, wherein:

47. R ９ is substituted with 3 to 5 -OH ２ -C ６ 47. The compound according to any one of claims 35 to 46, or a pharma- ceutically acceptable form thereof, wherein: - alkyl.

48. R ９ but:

48. The compound according to any one of claims 35 to 47, which is:

49. Z ３ But, C ６ -C １０ -aryl or 5-10 membered heteroaryl, where 1-4 heteroaryl members are independently selected from N, O and S; n3 is 1 or 2; R ８ But, C １ -C １０ - alkyl; R ９ is C substituted with 1 to 5 -OH ２ -C ６ - alkyl.

50. 36. A compound according to claim 1 or 35, which is selected from the table below, or a pharma- ceutically acceptable version thereof.

51. 51. A pharmaceutical composition comprising a compound according to any one of claims 1 to 50, or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable carrier.

52. 51. A pharmaceutical composition for selectively increasing proliferation of cuboidal alveolar type 2 (AEC2) cells in a subject in need thereof or for restoring decreased proliferation of AEC2 cells in a subject in need thereof, comprising a compound according to any one of claims 1 to 50 or a pharma- ceutically acceptable salt thereof.

53. 51. A pharmaceutical composition for inhibiting dipeptidyl peptidase IV (DPP4) in a subject in need thereof, comprising a compound according to any one of claims 1 to 50, or a pharma- ceutically acceptable salt thereof.

54. 51. A pharmaceutical composition for treating a pulmonary disease in a subject, comprising a compound according to any one of claims 1 to 50, or a pharma- ceutically acceptable salt thereof.

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