Small molecule regulators of alveolar type 2 cell proliferation for the treatment of lung 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 priority to U.S. Provisional Patent Application No. 63 / 139,956, filed on 21 January 2021, and this application is incorporated in its entirety as if it were fully described herein. [Background technology]

[0002] Pharmacological stimulation of lower airway repair has great potential for treating various conditions in which alveolar destruction and maladaptive remodeling are the cause of disease. The alveoli, the main units of gas exchange in mammals, are composed of two epithelial cell types: large squamous alveolar type 1 cells (AEC1) that provide surface area for gas exchange, and cuboidal alveolar type 2 cells (AEC2) that secrete surfactant. 1 Furthermore, AEC2 has been identified as a primary progenitor cell type responsible for the regeneration of alveolar epithelium. 2 AEC2 clonally proliferates throughout adulthood, dividing asymmetrically to give rise to AEC1 and AEC2. 2 It has been further demonstrated that idiopathic pulmonary fibrosis (IPF) is caused by the depletion of AEC2 stem cell capacity. 3 When AEC2 proliferation decreases, the alveolar basement membrane peels off, 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 through treatment with exogenous factors (IL-6 or hyaluronic acid) inhibits disease severity in mouse models of IPF. 4 In addition to IPF, acute respiratory distress syndrome (ARDS) (acute loss of alveolar epithelial barrier function) is caused by damage to AEC2 cells and insufficient repair growth by AEC2 cells. 5 . [Overview of the project]

[0003] In various embodiments, the present disclosure provides compounds useful, for example, for promoting the specific proliferation of AEC2s compared with other cell types in the lung. In some embodiments, the compound is of formula (I) or a pharmaceutically acceptable salt thereof.

Chemical

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

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

[0006] X 1 is selected from -O-, -S-, -S(O)-, S(O)2- and -NH-. L 1B is C2-C 12 alkyl, wherein one or more -CH2- groups may be independently substituted with a moiety selected from -O-, -C(O)- and -NH-. Z 1 is selected from H, C6-C 10 aryl and 5- to 10-membered heteroaryl, wherein 1 to 4 heteroaryl ring members are independently selected from N, O and S.

[0007] Subscript m1 is an integer that is 0 when Z 1 is H, and 1 when Z 1 is other than H. Subscript n1 is an integer selected from 0, 1, 2, and 3.

[0008] R 1 is selected from H, C1-C 10 alkyl and -C1-C 10 alkyl-(C6-C 10 aryl), and may be substituted with 1 to 6 -OH groups. R 2 is C1-C 10 alkyl substituted with 1 to 6 -OH groups.

[0009] Alternatively, according to further embodiments, the compound is formula (II) or a pharmaceutically acceptable salt thereof: [ka]

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

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

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

[0013] R 5a , R 5b , R 5c and R 5d H, C1-C6-alkyl, halo, -NR A R B (R A and R B H and C1-C 10 -Selected independently of alkyl, -C(O)OH, -B(OH)2, -C(O)NR A R B , -C(O)OR A and -C(O)-L 2 -Z 2 -[(CH2) n2 -NR 6 R 7 ] m2 It is selected independently of L. 2 C2-C 12 -Alkyl, and one or more -CH2- groups may be independently substituted by moieties selected from -O-, -C(O)-, and -NH-. 2 H, C6-C 10- Selected from aryl and 5-10 member heteroaryls (1-4 heteroaryl members are independently selected from N, O, and S). 5a , R 5b , R 5c and R 5d At least one of them is not H.

[0014] R 6 H, C1-C 10 -alkyl and -C1-C 10 -alkyl-(C6-C 10 Selected from -aryl groups, and may be substituted with 1 to 6 -OH groups. 7 This is a C1-C molecule substituted with 1 to 6 -OH groups. 10 -It is alkyl.

[0015] The subscript m2 is Z 1 If H, then it is 0, and Z 1 It is an integer that is 1 if it is not H. The subscript n2 is an integer selected from 0, 1, 2, and 3.

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

[0017] Alternatively, according to further embodiments, the compound is of formula (III) or a pharmaceutically acceptable salt thereof: [ka]

[0018] In formula (III), X 3 L is -O- or -NH-. 3 is a bond or C2-C 12 -Alkyl, and one or more -CH2- groups may be independently substituted by moieties selected from -O-, -C(O)-, and -NH-. 3 H, -N3, C6-C 10-Selected from aryl, 5-10 membered heteroaryl (1-4 heteroaryl members independently selected from N, O, and S), and (3-14 membered heterocycloalkyl (1-4 ring members independently selected from N, O, and S). Heteroaryl and heterocycloalkyl may be substituted with 1-6 substituents selected from the group consisting of halo, NO2, OH, CN, and C1-C6-haloalkyl.

[0019] The subscript m3 is Z 3 It becomes 0 when H or -N3, and Z 3 It is an integer that is 1 if it is not 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 Selected from -aryl groups, and may be substituted with 1 to 6 -OH groups. 9 This is a C1-C molecule substituted with 1 to 6 -OH groups. 10 -It is alkyl. 10 Each R is a C1-C6 haloalkyl. 11 The element 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] The compounds disclosed herein do not include any of the following compounds: [Table 1] TIFF0007927730000005.tif212144TIFF0007927730000006.tif50148

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

[0024] This disclosure also provides, in one embodiment, a method for selectively increasing the proliferation of cuboidal alveolar type 2 (AEC2) cells in a subject requiring such treatment, or for restoring decreased AEC2 cell proliferation in a subject requiring such treatment. The method comprises administering a compound described herein or a pharmaceutically acceptable salt thereof to the subject.

[0025] In one embodiment, the Disclosure provides a method for inhibiting dipeptidyl peptidase IV (DPP4) in a subject where such inhibition is required. The method comprises administering a compound described herein or a pharmaceutically acceptable salt thereof to the subject.

[0026] Another embodiment of the present disclosure is a method for treating a lung disease in a subject suffering from it. The method comprises administering a compound described herein or a pharmaceutically acceptable salt thereof to a subject.

[0027] In various embodiments, compounds described herein or pharmaceutically acceptable salts thereof are also provided for use in selectively increasing the proliferation of cuboidal alveolar type 2 (AEC2) cells in subjects where such proliferation is required, or for restoring a decrease in AEC2 cell proliferation in subjects where such proliferation is required.

[0028] In further embodiments, the disclosure provides compounds described herein or pharmaceutically acceptable salts thereof for use in inhibiting dipeptidyl peptidase IV (DPP4) in subjects where such inhibition is required.

[0029] In further embodiments, the disclosure provides compounds described herein or pharmaceutically acceptable salts thereof for use in the treatment of lung diseases in affected subjects. [Brief explanation of the drawing]

[0030] [Figure 1] AEC2 growth-concentration curve for compound 46. [Figure 2] Comparison of mouse pharmacokinetics of liragliptin and compound 46 administered intratracheally (IT) at a dose of 2 mg / kg. [Figure 3A-3E] Compound 46 demonstrated efficacy in a mouse model of bleomycin-induced pulmonary fibrosis. (A) Body weight measurement and administration schedule used for intratracheal administration of compound 46 (0.5 mg / kg every 4 days). BALF (B), fibrosis area measurement (C), modified Ashcroft score (D), and representative Masson trichrome stained histological slides at the end of the study (E);**, P<0.005;***, P<0.0005. [Figure 4A-4D] Compound 46 shows a synergistic effect in combination with the standard IPF drug nintedanib in a mouse model of bleomycin-induced pulmonary fibrosis. (A) Legend of treatment 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 trichrome stained histological slides at the end of the study (D). *, P<0.05; ***; P<0.005; ***, P<0.0005; NS = not statistically significant. [Figures 5A-5C] Compound 46 selectively increased AEC2 in mice. (A) UMAP plot showing multiple cell populations identified in mouse lung, highlighting the important population of interest. (B) UMAP plot showing which cells express transcriptional profiles consistent with the proliferation state. The other populations besides the proliferating AEC2 are immune cells. (C) Quantification of all proliferating cells at the indicated time point after treatment with compound 46. [Modes for carrying out the invention]

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

[0032] definition "Alkyl" refers to a linear or branched hydrocarbyl containing 1 to approximately 20 carbon atoms. For example, alkyls can have 1 to 10 carbon atoms or 1 to 6 carbon atoms. Exemplary alkyls include linear alkyl groups such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, and dodecyl, as well as branched isomers of linear alkyl groups, for example, but not limited to, -CH(CH3)2, -CH(CH3)(CH2CH3), -CH(CH2CH3)2, -C(CH3)3, -C(CH2CH3)3, -CH2CH(CH3)2, and -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、 This includes -CH(CH3)CH(CH3)CH(CH3)2, etc. Therefore, alkyl groups include primary alkyl groups, secondary alkyl groups, and tertiary alkyl groups. Alkyl groups may be unsubstituted or substituted with one or more substituents as described herein.

[0033] The term "haloalkyl" is an alkyl group as defined herein that is substituted with 1, 2, 3, 4, 5, or 6 halos. An example haloalkyl group is -CF3.

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

[0035] "Alkyne" or "alkynyl" refers to a linear or branched 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-pentine, 2-pentine, 1-hexine, 2-hexine, 3-hexine, 1-heptine, 2-heptine, 3-heptine, 1-octin, 2-octin, 3-octin, and 4-octin. The alkynyl group may be unsubstituted or substituted with one or more substituents as described herein.

[0036] When used alone or as part of another term, "aryl" refers to a fused or fused carbocyclic aromatic group having a specified number of carbon atoms, or, if no number is specified, up to 14 carbon atoms, e.g., C6-C6 14 -Aryl or C6-C 10 - Represents aryl. Exemplary aryl groups include phenyl, naphthyl, biphenyl, phenantrenyl, and naphthacenyl (see, for example, Table 7-2 in Lang's Handbook of Chemistry (Dean, JA, ed.), 13th edition). An exemplary aryl is phenyl. "Aryl" may be fused with a cycloalkyl ring as defined herein. The 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 this disclosure containing an N or S atom may be oxidized to the corresponding N-oxide, sulfoxide, or sulfone compound.

[0038] "Heteroaryl" refers to a monocyclic aromatic ring structure containing 5 to 10 ring atoms, e.g., 5 or 6 ring atoms, either alone or in combination with any other part described herein, or a bicyclic aromatic group having 8 to 10 atoms and containing one or more heteroatoms independently selected from the group consisting of O, S, and N, e.g., 1 to 4, 1 to 3, or 1 to 2. Heteroaryls are also intended to contain oxidation of S or N, e.g., sulfinyl, sulfonyl, and N-oxides of tertiary ring nitrogen. The carbon or heteroatom is a bonding site in the heteroaryl ring structure, thereby creating a stable compound. Examples of heteroaryl groups include, but are not limited to, pyridinyl, pyridadinyl, pyrazinyl, quinaoxalyl, indolidinyl, benzo[b]thienyl, quinazolinyl, prinyl, indolyl, quinolinyl, pyrimidinyl, pyrrolyl, pyrazolyl, oxazolyl, thiazolyl, thienyl, isoxazolyl, oxathiadiazolyl, isothiazolyl, tetrazolyl, imidazolyl, triazolyl, furanyl, benzofuryl, and indolyl. Heteroaryl groups may be unsubstituted or substituted with one or more substituents described herein.

[0039] A "heterocycloalkyl" is a saturated or partially unsaturated non-aromatic monocyclic, bicyclic, tricyclic, or polycyclic ring system having 3 to 14 atoms, for example, 3 to 6 atoms, of which 1 to 3 carbon atoms in the ring are replaced by heteroatoms of O, S, or N, and which may be spirocondensed. Heterocycloalkyls may be condensed with 5 to 6-membered aryl or heteroaryl rings and include S or N oxides, such as sulfinyl, sulfonyl, and N-oxides of tertiary ring nitrogen. The bonding sites of the heterocycloalkyl ring are located on carbon or heteroatoms, resulting in the retention of a stable ring. Examples of heterocycloalkyls include, but are not limited to, morpholino, tetrahydrofuranyl, dihydropyridinyl, piperidinyl, pyrrolidinyl, piperazinyl, dihydrobenzofuryl, and dihydroindolyl. Heterocycloalkyls may be unsubstituted or substituted with one or more substituents as described herein.

[0040] The terms "nitrile" and "cyano" can be used interchangeably and refer to the -CN group bonded to the carbon atoms of heteroaryl, aryl, and heterocycloalkyl rings.

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

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

[0043] The substituent -CO2H is a bioisosteric substitution, for example [ka]

[0044] These may be replaced with, for example, where R is defined as R as defined herein. A It has the same definition. For example, see page 203 of The Practice of Medicinal Chemistry (Academic Press: New York, 1996).

[0045] The compounds described herein may exist in a variety of isomeric forms, including configurational isomers, geometric isomers, and conformational isomers, for example, 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 in this disclosure, including tautomers of the compounds. The compounds in this disclosure may also exist in chain or cyclized forms. In some cases, one or more cyclized forms may result from the loss of water. The specific composition of chain and cyclized forms may depend on how the compounds are isolated, stored, or administered. For example, a compound may exist primarily in chain form under acidic conditions but may be cyclized under neutral conditions. All forms are included in this disclosure.

[0046] Some of the compounds described herein may have chiral centers and therefore may exist in various enantiomer and diastereomer forms. The compounds described herein may be in the form of optical isomers or diastereomers. Accordingly, this disclosure encompasses the compounds described herein in the form of optical isomers, diastereomers, and mixtures thereof (including racemic mixtures) and their uses. Optical isomers of the compounds of this disclosure can be obtained by known techniques, such as asymmetric synthesis, chiral chromatography, pseudo-mobile bed techniques, or chemical separation of stereoisomers using optically active resolving agents.

[0047] Unless otherwise specified, the term “stereoisomer” means one stereoisomer of a compound and substantially no other stereoisomers of that compound. Thus, a stereoisomerically pure compound having one chiral center substantially does not contain the opposite enantiomer of the compound. A stereoisomerically pure compound having two chiral centers substantially does not contain the other diastereomer of the compound. A typical stereoisomerically pure compound contains more than 80% by weight of one stereoisomer of the compound and less than 20% by weight of the other stereoisomer of the compound, for example, more than 90% by weight of one stereoisomer of the compound and less than 10% by weight of the other stereoisomer of the compound, or more than 95% by weight of one stereoisomer of the compound and less than 5% by weight of the other stereoisomer of the compound, or more than 97% by weight of one stereoisomer of the compound and less than 3% by weight of the other stereoisomer of the compound, or more than 99% by weight of one stereoisomer of the compound and less than 1% by weight of the other stereoisomer of the compound. The stereoisomers described above can be considered as compositions comprising two stereoisomers present in the respective weight percentages described herein.

[0048] In the event of any discrepancy between a shown structure and the name given to it, the shown structure prevails. Furthermore, if the stereochemistry of a structure or part of a structure is not indicated, for example, by a thick or dashed line, the structure or part of a structure should be interpreted as encompassing all of its stereoisomers. However, in some cases where multiple chiral centers are present, the structure and name may be expressed as a single enantiomer to help describe the relative stereochemistry. Those skilled in organic synthesis will know whether compounds are prepared as a single enantiomer from the methods used to prepare them.

[0049] As used herein, unless otherwise specified, the term “compound” is inclusive in that it includes a compound or its pharmaceutically acceptable salts, stereoisomers, and / or tautomers. Thus, for example, the compounds of this disclosure include pharmaceutically acceptable salts of the tautomers of that compound.

[0050] In this specification, “pharmaceutically acceptable salt” refers to a pharmaceutically acceptable organic or inorganic acid or base salt of a compound described herein. Typical pharmaceutically acceptable salts include, for example, alkali metal salts, alkaline earth salts, ammonium salts, water-soluble and water-insoluble salts, such as acetates, amsonates (4,4-diaminostilbene-2,2-disulfonate), benzenesulfonates, benzoates, bicarbonates, bisulfates, tartrates, borates, bromides, butyrates, calcium, calcium edetate, cansylates, carbonates, chlorides, citrates, and klavulariae. Clavulariate, dihydrochloride, edetate, edisylate, estrate, esylate, fiunarate, gluceptate, gluconate, glutamate, glycolylarsanilate, hexafluorophosphate, hexylresorcinate, hydravamin, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, Isothionate, lactate, lactobionate, laurate, malate, maleate, mandelate, mesylate, methyl bromide, methylnitrate, 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 (einbon Examples include phosphates, pantothenates, phosphates / diphosphates, picrates, polygalacturonates, propionates, p-toluenesulfonates, salicylates, stearates, basic acetates, succinates, sulfates, sulfosaliculates, suramates, tannates, tartrates, teoclates, tosylates, triethiozides, and valersates. A pharmaceutically acceptable salt may have multiple charged atoms in its structure. In this example, a pharmaceutically acceptable salt may have multiple counterions.Therefore, a pharmaceutically acceptable salt may have one or more charged atoms and / or one or more counterions.

[0051] The terms “to treat,” “to treat,” and “treatment” refer to the improvement or elimination of a disease or symptoms associated with a disease. In certain embodiments, such terms refer to minimizing the progression or worsening of a disease as a result of administering one or more prophylactic or therapeutic agents to a patient having such a disease.

[0052] The terms “prevent,” “prevention,” and “prevention” refer to the prevention of the onset, recurrence, or progression of a disease in a patient, resulting from the administration of a preventive or therapeutic agent.

[0053] The term "effective dose" refers to the amount of the 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 the disease. Furthermore, the therapeutic effective dose of the compounds described herein means the amount of the 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 relation to the compounds described herein, this term may include amounts that improve overall treatment, reduce or avoid the symptoms or causes of the disease, or enhance or synergistically enhance the therapeutic efficacy of another therapeutic agent.

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

[0055] "Inhibitor" means a compound that prevents or reduces the expression, catalytic activity, and / or localization (i.e., local concentration) of DPP4.

[0056] Compound As generally described above, the present disclosure provides a compound of formula (I) or a pharmaceutically acceptable salt thereof.

Chemical Formula

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

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

[0059] X 1 is selected from -O-, -S-, -S(O)-, S(O)2- and -NH-. L 1B is C2-C 12 -alkyl, wherein one or more -CH2- groups may be independently substituted with a moiety selected from -O-, -C(O)- and -NH-. Z 1 is selected from H, C6-C 10 -aryl and 5- to 10-membered heteroaryl, wherein 1 to 4 heteroaryl ring atoms are independently selected from N, O and S.

[0060] Subscript m1 is Z 1 is 0 when is H, and Z 1 is an integer that is 1 when is other than H. Subscript n1 is an integer selected from 0, 1, 2 and 3.

[0061] R 1 is selected from H, C1-C 10 -alkyl and -C1-C 10 -alkyl-(C6-C 10 -aryl), and may be substituted with 1 to 6 -OH groups.

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

Chemical Structure

[0063] In various embodiments, L 1A is -NHCH2-. In other embodiments, 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- to 10-membered heteroaryl, wherein 1 to 4 heteroaryl ring members are independently selected from N, O and S. According to one embodiment, an example of C6-C 10 -aryl is phenyl. According to another embodiment, an example of heteroaryl 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 embodiments described herein, R 1 is C1-C 10 -alkyl optionally substituted with 1 to 6 -OH groups. For example, in an exemplary embodiment, R 1 is C1-C6-alkyl.

[0068] In further embodiments, R may be combined with any other embodiments described herein. 2 is a C2-C6 alkyl group substituted with 1 to 5 -OH groups, or R 2 R is a C2-C6 alkyl group substituted with 3 to 5 -OH groups. In exemplary embodiments, R 2 The following applies: [ka]

[0069] Further embodiments provide compounds of formula (I), wherein, Z 1 C6-C 10 -The aryl or 5-10 member heteroaryl (1-4 heteroaryl members are independently selected from N, O, and S); n1 is either 1 or 2; R 1 C1-C 10 -It is alkyl; R 2 These are C2-C6-alkyl groups substituted with 1 to 5 -OH groups.

[0070] In some embodiments, there are no single bonds represented by "---". In other embodiments, they are present to form a condensed cyclopropyl ring. These embodiments are each represented by the following structures. [ka]

[0071] As described herein, Part L 1B C2-C 12-It is alkyl. In some embodiments, 1, 2, 3, 4, or 5 -CH2- groups are replaced. It should be understood that, according to chemical principles, substitutions only form stable compounds, such as when substitution occurs at adjacent -CH2- groups. 1B Examples include the following: [ka]

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

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

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

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

[0076] R 5a , R 5b , R 5c and R 5d H, C1-C6-alkyl, halo, -NR A R B (R A and R B H and C1-C 10 -Selected independently of alkyl, -C(O)OH, -B(OH)2, -C(O)NR A R B , -C(O)OR A and -C(O)-L 2 -Z 2 -[(CH2)n2 -NR 6 R 7 ] m2 It is selected independently of L. 2 C2-C 12 -Alkyl, and one or more -CH2- groups may be independently substituted by moieties selected from -O-, -C(O)-, and -NH-. 2 H, C6-C 10 - Selected from aryl and 5-10 member heteroaryls (1-4 heteroaryl members are independently selected from N, O, and S). 5a , R 5b , R 5c and R 5d At least one of them is not H.

[0077] In addition, if W is CH, then R 5a and R 5d It 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 Selected from -aryl groups, and may be substituted with 1 to 6 -OH groups. 7 This is a C1-C molecule substituted with 1 to 6 -OH groups. 10 -It is alkyl. In various embodiments, R 7 It is substituted with 1, 2, 3, 4, 5, or 6 -OH groups. Exemplary R 7 These are shown below, along with some examples of their diastereomers: [ka]

[0079] The subscript m2 is Z 1 If H, then it is 0, and Z 1 It is an integer that is 1 if it is not H. The subscript n2 is an integer selected from 0, 1, 2, and 3.

[0080] In one embodiment, W is CH. In another embodiment, W is N.

[0081] In various embodiments, R 5a , R 5b , R 5c and R 5d One, two, or three of them are H. In some embodiments, R 5b and R 5d Each of them is H. In some embodiments, R 5c is a halo, for example, chloroform. In yet another embodiment, R 5a is -C(O)OH or -C(O)OR A In further embodiments, R 5a is -C(O)-L 2 -Z 2 -[(CH2) n2 -NR 6 R 7 ] m2 These embodiments and their combinations are all intended.

[0082] R 5a , R 5b , R 5c and R 5d One example is R. 5a -C(O)-L 2 -Z 2 -[(CH2) n2 -NR 6 R 7 ] m2 In relation to the embodiment, a further embodiment is Z 2 Provided as phenyl or triazolyl. In exemplary embodiments, Z 2 It is triazolyl.

[0083] In additional embodiments, and potentially in combination with any other embodiments described herein, n2 is 1 or 2.

[0084] In further embodiments, R may be combined with any other embodiments described herein. 6 This C1-C group may be substituted with 1 to 6 -OH groups. 10 -It is alkyl. In one embodiment, R 6 It is a C1-C6 alkyl group.

[0085] In additional embodiments, R may be combined with any other embodiments described herein as applicable. 7 is a C2-C6 alkyl group substituted with 1 to 5 -OH groups or 3 to 5 -OH groups. For example, in an exemplary embodiment, R 7 The following is true: [ka]

[0086] In further embodiments, the present 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 C6-C 10 -The aryl or 5-10 member heteroaryl (1-4 heteroaryl members are independently selected from N, O, and S); n2 is either 1 or 2; R 6 C1-C 10 -It is alkyl; R 7 These are C2-C6-alkyl groups substituted with 1 to 5 -OH groups.

[0087] As described herein, in various embodiments, part L 2is C2-C 12 -alkyl, wherein one or more -CH2- groups may be independently substituted with -O-, -C(O)- and -NH-. In some embodiments, 1, 2, 3, 4 or 5 -CH2- groups are substituted. It should be understood that, according to chemical principles, substitution only forms stable compounds, such as when substitution occurs at adjacent -CH2- groups. Examples of L 2 include the following.

Chemical Formula

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

[0089] , optionally in combination with any other embodiments, are various embodiments wherein Z 2 is C6-C 10 -aryl or 5- to 10-membered heteroaryl, wherein 1 to 4 heteroaryl ring members are independently selected from N, O and S. In one embodiment, Z 2 is C6-C 10 -aryl such as phenyl.

[0090] In a further embodiment, the compound is one of formula (III), or a pharmaceutically acceptable salt thereof:

Chemical Formula

[0091] In formula (III), X 3 is -O- or -NH-. L 3 is a bond or C2-C 12 -alkyl, wherein one or more -CH2- groups may be independently substituted with a moiety selected from -O-, -C(O)- and -NH-. Z 3 is H, -N3, C6-C 10-Selected from aryl, 5-10 membered heteroaryl (1-4 heteroaryl members independently selected from N, O, and S), and (3-14 membered heterocycloalkyl (1-4 ring members independently selected from N, O, and S). Heteroaryl and heterocycloalkyl may be substituted with 1-6 substituents selected from the group consisting of halo, NO2, OH, CN, and C1-C6-haloalkyl.

[0092] The subscript m3 is Z 3 It becomes 0 when H or -N3, and Z 3 It is an integer that is 1 if it is not 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 Selected from -aryl groups, and may be substituted with 1 to 6 -OH groups. 9 This is a C1-C molecule substituted with 1 to 6 -OH groups. 10 -It is alkyl. 10 Each R is a C1-C6 haloalkyl. 11 The element 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 In other embodiments, X 3 It is -NH-.

[0096] As described herein, Part L 3 C2-C12 -It is alkyl. In some embodiments, it is partially L 3 C5-C 12 -alkyl, C5-C 10 -Alkyl, C7-C 12 -alkyl, C8-C 12 -alkyl or C9-C 12 -It is alkyl. In some embodiments, depending on the circumstances, 1, 2, 3, 4, or 5 -CH2- groups are replaced in combination with any embodiment described herein. It should be understood that, according to chemical principles, substitutions form only stable compounds, such as when substitution occurs at adjacent -CH2- groups. 3 Examples include: [ka]

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

[0098] In other embodiments, Z 3 C6-C 10 - Aryl or 5-10 member heteroaryl (1-4 heteroaryl members are independently selected from N, O, and S). Exemplary Z 3 It is phenyl or triazolyl.

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

[0100] In further embodiments, R may be combined with any other embodiments described herein. 8 This C1-C group may be substituted with 1 to 6 -OH groups. 10 -It is alkyl. In an exemplary embodiment, R 8 It is a C1-C6 alkyl group.

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

Chemical Formula

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

[0103] Notwithstanding the general definitions provided herein, it is to be understood that the compound does not include any of the following compounds: Table 2 TIFF0007927730000021.tif210135

[0104] In yet additional embodiments, the present disclosure provides specific compounds or pharmaceutically acceptable salts thereof shown in the following Tables 1 to 4. Table 3 TIFF0007927730000023.tif183164TIFF0007927730000024.tif160163 Table 4 Table 5 TIFF0007927730000027.tif70163

[0105] Pharmaceutical composition This disclosure also provides pharmaceutical compositions comprising a therapeutically effective amount of one or more of the compounds described herein, or pharmaceutically acceptable salts, stereoisomers, and / or tautomers thereof, mixed with a pharmaceutically acceptable carrier. In some embodiments, the composition further comprises one or more additional therapeutic agents, pharmaceutically acceptable excipients, diluents, adjuvants, stabilizers, emulsifiers, preservatives, colorants, buffers, and flavoring agents, in accordance with acceptable practices of pharmaceutically formulation.

[0106] In one embodiment, the pharmaceutical composition comprises a compound selected from any of the compounds shown in any table disclosed herein or their pharmaceutically acceptable salts, stereoisomers and / or tautomers, and a pharmaceutically acceptable carrier.

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

[0108] The “therapeutic effective dose” of the compound administered or its pharmaceutically acceptable salts, stereoisomers, and / or tautomers is the minimum amount required to regenerate AEC2 cell proliferation, inhibit DPP4, or both, depending on such considerations. Such a dose may be less than the amount that is toxic to normal cells or the overall subject. Generally, the initial therapeutic effective dose of the compound of the Disclosure administered (or its pharmaceutically acceptable salts, stereoisomers, or tautomers) is in the range of about 0.01 to about 200 mg per kg of patient body weight or about 0.1 to about 20 mg / day, with a typical initial range of about 0.3 to about 15 mg / kg / day. Oral unit dosage forms such as tablets and capsules may contain about 0.1 mg to about 1000 mg of the compound of the Disclosure (or its pharmaceutically acceptable salts, stereoisomers, or tautomers). In another embodiment, such a dosage form contains about 50 mg to about 500 mg of the compound of the Disclosure (or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof). In yet another embodiment, such a dosage form contains about 25 mg to about 200 mg of the compound of the Disclosure (or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof). In yet another embodiment, such a dosage form contains about 10 mg to about 100 mg of the compound of the Disclosure (or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof). In yet another embodiment, such a dosage form contains about 5 mg to about 50 mg of the compound of the Disclosure (or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof). In any of the embodiments described above, the dosage form may be administered once or twice daily.

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

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

[0111] In another embodiment, the disclosure also includes pharmaceutical compositions suitable for single-dose administration, comprising the compounds of this disclosure or their pharmaceutically acceptable stereoisomers, salts, or tautomers, and pharmaceutically acceptable carriers.

[0112] The compositions of this disclosure suitable for oral use can be prepared according to any method known in the art for the manufacture of pharmaceutical compositions. For example, liquid formulations of the compounds of this disclosure contain one or more agents selected from the group consisting of sweeteners, flavoring agents, coloring agents and preservatives in order to provide a pharmaceutically palatable preparation of the compound.

[0113] For tablet compositions, the compounds of this disclosure, mixed with non-toxic, pharmaceutically acceptable excipients, are used in the manufacture of 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; granulators and disintegrants such as corn starch or alginic acid; binders such as starch, gelatin, or acacia; and lubricants such as magnesium stearate, stearic acid, or talc. Tablets may or may not be coated, or they may be coated by known coating techniques to delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained therapeutic effect over a desired period. For example, time-delaying materials such as glyceryl monostearate or glyceryl distearate can be used.

[0114] Formulations for oral use may also be provided as rigid 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 oily medium, such as peanut oil, liquid paraffin, or olive oil.

[0115] In the case of aqueous suspensions, the compounds of this disclosure are mixed with excipients suitable for maintaining a stable suspension. Examples of such excipients include, but are not limited to, sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, tragacanth gum, and gum arabic.

[0116] The oral suspension may also contain a dispersant or wetting agent, such as a natural phosphatide, such as lecithin, or a condensation product of an alkylene oxide and a fatty acid, such as polyoxyethylene stearate, or a condensation product of an ethylene oxide and a long-chain aliphatic alcohol, such as heptadecaethyleneoxycetanol, or a condensation product of an ethylene oxide and a partial ester derived from a fatty acid and hexitol, such as polyoxyethylene sorbitol monooleate, or a condensation product of an ethylene oxide and a partial ester derived from a fatty acid and hexitol anhydride, such as polyethylene sorbitan monooleate. The aqueous suspension may also contain one or more preservatives, such as ethyl p-hydroxybenzoate or n-propyl p-hydroxybenzoate, one or more colorants, one or more flavoring agents, and one or more sweeteners, such as sucrose or saccharin.

[0117] The oily suspension can be formulated by suspending the compounds of this disclosure in a vegetable oil, such as peanut oil, olive oil, sesame oil, or coconut oil, or in a mineral oil, such as liquid paraffin. The oily suspension may contain a thickener, such as beeswax, hard paraffin, or cetyl alcohol.

[0118] By adding the sweeteners and flavorings described above, an oral formulation with a pleasant mouthfeel can be provided. These compositions can be preserved by adding antioxidants such as ascorbic acid.

[0119] Dispersible powders and granules suitable for the preparation of aqueous suspensions by the addition of water are provided, which are compounds of the present disclosure mixed with a dispersant or wetting agent, a suspending agent and one or more preservatives. Suitable dispersants or wetting agents and suspending agents are exemplified by those already described above. Additional excipients, such as sweeteners, flavoring agents and coloring agents, may also be present.

[0120] The pharmaceutical compositions of this disclosure may be in the form of oil-in-water emulsions. The oil phase may be a vegetable oil, such as olive oil or peanut oil, or a mineral oil, such as liquid paraffin, or a mixture thereof. Suitable emulsifiers may be natural gums, such as gum arabic or tragacanth gum, natural phosphatides, such as soy, lecithin, and esters or partial esters derived from fatty acids and hexitol, anhydrides, such as sorbitan monooleate, and condensation reaction products of the partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate. The emulsion may also contain sweeteners and flavorings.

[0121] Syrups and elixirs can be formulated with sweeteners, such as glycerol, propylene glycol, sorbitol, or sucrose. Such formulations may also contain lubricants, preservatives, and flavorings and colorings. Pharmaceutical compositions may be in the form of sterile injections, aqueous suspensions, or oily suspensions. These suspensions can be formulated according to known techniques using the appropriate dispersants or wetting and suspending agents described above. Sterile injection preparations may also be sterile injection solutions or suspensions in non-toxic, parenterally acceptable diluents or solvents, such as solutions in 1,3-butanediol. Acceptable vehicles and solvents that can be used include water, Ringer's solution, and isotonic sodium chloride solutions. Furthermore, sterile fixative oils have been conventionally used as solvents or suspension media. For this purpose, any low-irritation fixative oil, including synthetic monoglycerides or diglycerides, can be used. In addition, fatty acids such as oleic acid are used in injection preparations.

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

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

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

[0125] The advantage of the compounds in this disclosure lies in their ability to modulate the regenerative capacity of AEC2 cells. For this property, drugs approved for IPF inhibit the activation and proliferation of pulmonary fibroblasts and myofibroblasts, which are the source of scar tissue in diseased lungs. In contrast, the compounds in this disclosure promote alveolar repair by directly targeting the source of disease in IPF (which is the ineffective self-regeneration of damaged AEC2 cells). Therefore, the AEC2-targeting compounds in this disclosure offer further disease-modifying effects, either as monotherapy or in combination with approved IPF drugs (e.g., pirfenidone).

[0126] Thus, the present disclosure provides methods for selectively increasing the proliferation of cuboidal alveolar type 2 (AEC2) cells in subjects requiring such proliferation, or for restoring decreased AEC2 cell proliferation in subjects requiring such proliferation. The methods include administering the compounds disclosed herein or pharmaceutically acceptable salts thereof to the subject.

[0127] Another embodiment also provides a method for inhibiting dipeptidyl peptidase IV (DPP4) in subjects where it is needed. The method involves administering a compound disclosed herein or a pharmaceutically acceptable salt to a subject.

[0128] This disclosure provides, in various embodiments, methods for treating subjects suffering from a disease or condition whose pathogenesis derives from epithelial degeneration, maladaptive remodeling, and / or ineffective self-regeneration of damaged AEC2 cells. In some embodiments, the disease is a lung disease or pulmonary condition.In further embodiments, the disease or condition may include idiopathic pulmonary fibrosis (IPF), acute respiratory distress syndrome (ARDS), chronic obstructive pulmonary disease (COPD), emphysema, silicosis, asbestosis, pneumoconiosis, aluminum lung, bauxite fibrosis, beryllium poisoning, siderosis, tin pneumonitis, talc lung, Labrador lung (mixed pneumoconiosis), sarcoidosis, hypersensitivity pneumonitis (HP) / exogenous allergic alveolitis (EAA), chronic bronchitis, desquamative interstitial pneumonia (DIP), respiratory bronchiolitis-associated interstitial lung disease (RBILD), acute interstitial pneumonia (AIP), and nonspecific interstitial pneumonia (NS). IP), Idiopathic organizing pneumonia (COP = Idiopathic 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, Idiopathic organizing pneumonia (COP), Cystic fibrosis (CF), E-cigarette or vaping-related 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 syndrome, idiopathic pulmonary hemophilia, alveolar hemorrhage syndrome of unknown cause, alveolar hemorrhage syndrome of confirmed cause, sporadic pulmonary lymphangioleiomyoma (S-LAM), pulmonary lymphangioleiomyoma in tuberous sclerosis (TSC-LAM), 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 feeder's lung disease). This includes all causes other than parental lung disease, pulmonary arteriovenous malformations (HHT) in hereditary hemorrhagic telangiectasia, interstitial lung disease in systemic sclerosis, interstitial lung disease in rheumatoid arthritis, interstitial lung disease in idiopathic inflammatory myopathy (polymyositis, dermatomyositis, anti-synthetase syndrome), interstitial lung disease in Sjögren's syndrome, interstitial lung disease in mixed connective tissue disease (MCTD), interstitial lung disease in overlapping syndromes, interstitial lung disease in undifferentiated connective tissue disease, and selected from (non-transplant patients) bronchiolitis obliterans.

[0129] In other embodiments, the disease is selected from inflammatory diseases, as well as 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 diabetic 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, diastolic heart failure (heart failure with preserved ejection fraction), atrial septal defect, atrioventricular septal defect, and aorta. Stenosis, double outlet right ventricle, transposition of the great arteries, Ebstein malformation, hypoplastic left heart syndrome, aortic interruption, 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 disease, IgA vasculitis, IgA nephropathy, lupus nephritis, adult nephrotic syndrome, childhood nephrotic syndrome, hemolytic uremic syndrome, medullary cavernous nephropathy, renal malformation, renal artery stenosis, renovascular hypertension, renal tubular acidosis, Alport syndrome, Wenger granulomatosis, Alagille syndrome, cystitis Gnosis, Fabry disease, focal segmental glomerulosclerosis (FSGS), glomerulonephritis, atypical hemolytic uremic syndrome (aHUS), hemolytic uremic syndrome (HUS), Henoch-Schönlein purpura, IgA nephropathy (Berger's disease), interstitial nephritis, minimal change syndrome, nephrotic syndrome, thrombotic thrombocytopenic purpura (TTP), granulomatosis with polyangiitis (GPA), eczema, psoriasis, cellulitis, impetigo, atopic dermatitis, epidermolysis bullosa, lichen sclerosing, ichthyosis, vitiligo, acropeeling skin syndrome, Blau syndrome, primary cutaneous amyloidosis, skin Skin abscess, pressure ulcer, blepharitis, furuncleosis, full-thickness or partial-thickness burns, capillary vasculitis, dermatitis, cellulitis, corneal abrasions, corneal erosions, xerosis, lichen planus, chronic lichen simplex, venous ulcers (stasis ulcers), 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), Barlow's disease, bullous pemphigoid,Celiac disease, chronic relapsing multiple osteomyelitis (CRMO), Churg-Strauss syndrome (CSS) or eosinophilic granulomatosis (EGPA), bullous pemphigoid scarring, Cogan syndrome, cold agglutinin disease, coxsackie myocarditis, CREST syndrome, dermatitis herpetiformis, dermatomyositis, Devic's disease (neuromyelitis optica), lupus discoid, 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-Schenlein Purpura (HSP), herpes zoster of pregnancy or bullous pemphigoid of pregnancy (PG), hypogammaglobulinemia, IgG4-related sclerosing disease, immune thrombocytopenic purpura (ITP), inclusion body myositis (IBM), Lambert-Eaton syndrome, leukocytoclastic vasculitis, linear IgA disease (LAD), microscopic polyangiitis (MPA), mixed connective tissue disease (MCTD), Mollen's ulcer, Mukka-Habermann syndrome, multifocal motor neuropathy (MMN) or MMNCB, multiple sclerosis, myasthenia gravis, myositis, narcolepsy, neonatal lupus, neuromyelitis optica, neutropenia, ocular scarring Pemphigus, optic neuritis, relapsing rheumatoid arthritis (PR), PANDAS, paraneoplastic cerebellar degeneration (PCD), paroxysmal nocturnal hemoglobinuria (PNH), Parry-Romberg disease, squamous cellulitis (peripheral uveitis), Personage-Turner syndrome, pemphigus, peripheral neuropathy, perivenosis-induced encephalomyelitis, pernicious anemia (PA), POEMS syndrome, polyarteritis nodosa, polysomnogenesis imperfecta 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 leg syndrome (RLS), retroperitoneal fibrosis, rheumatic fever, rheumatoid arthritis, sarcoidosis, Schmidt syndrome, scleritis, scleroderma, Sjögren's syndrome, sperm and testicular autoimmunity, stiff person syndrome (SPS), subacute bacterial endocarditis (SBE), Suzak syndrome, sympathetic ophthalmitis (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,This includes a selection of conditions from lysosomal acid lipase deficiency (LAL-D), hepatic cysts, liver cancer, neonatal jaundice, non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, primary biliary cholangitis (PBC), progressive familial intrahepatic cholestasis (PFIC), osteoporosis, Paget's disease, osteonecrosis, osteoarthritis, low bone 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 prepared by procedures similar to those illustrated below.

[0139] [Examples] intermediate compound Synthesis of (2R,3R,4R,5S)-6-(hexyl(propa-2-in-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-pentaol [ka]

[0141] Raney Ni (1.2 g) was added 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) at room temperature. The reaction mixture was heated at 65 °C for 16 hours under H2 gas pressure (160 psi). After consumption of the starting materials, the reaction mixture was filtered through a Celite bed. The filtrate was concentrated under reduced pressure to obtain 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(propa-2-in-1-yl)amino)hexane-1,2,3,4,5-pentaol(Int-1): [ka]

[0143] (2R,3R,4R,5S)-6-(hexylamino)hexane-1,2,3,4,5-pentaol (1.0 g, 3.77 mmol) was stirred in THF (15 mL) to which 3-bromopropa-1-yin (80% solution in toluene, 0.84 mL, 5.65 mmol) was added, and the reaction mixture was stirred at 80°C for 16 hours. After consumption of the starting material, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain 0.5 g of (2R,3R,4R,5S)-6-(hexyl(propa-2-yin-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-cyanopyrrolidine-1-yl)-2-oxoethyl)amino)adamantan-1-ylmethanesulfonate(Int-2) [ka]

[0145] Step 1: tert-butyl(2-((S)-2-cyanopyrrolidine-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 equivalent) and (Boc)2O (165 g, 758 mmol, 1.15 equivalent) in THF (400 mL) and t-BuOH (400 mL) was degassed, purged three times with N2, and then the mixture was incubated at 70°C for 12 hours under an N2 atmosphere. The mixture was stirred. The solution was concentrated under reduced pressure, and MTBE (200 mL) was added. The mixture was filtered to obtain tert-butyl(2-((S)-2-cyanopyrrolidine-1-yl)-2-oxoethyl)((1S,3R,5S)-3-hydroxyadamantan-1-yl)carbamate (239 g, 592 mmol, yield 89.8%, purity 99.7%) as a white solid. TLC system: PE:EA(0:1);R f :0.40.

[0147] Process 2 (1R,3S,5S)-3-((tert-butoxycarbonyl)(2-((S)-2-cyanopyrrolidine-1-yl)-2-oxoethyl)amino)adamantan-1-ylmethanesulfonate (Int-2): [ka]

[0148] A solution of the compound tert-butyl(2-((S)-2-cyanopyrrolidine-1-yl)-2-oxoethyl)((1S,3R,5S)-3-hydroxyadamantan-1-yl)carbamate (230 g, 570 mmol, 1.00 equivalent) and N,N,N',N'-tetramethyl-1,6-hexanediamine (216 g, 1.25 mol, 2.20 equivalents) in DCM (2.30 L) was cooled to 0-10°C, and methanesulfonyl chloride (97.9 g, 855 mmol, 1.50 equivalents) was added at 0-10°C. The reaction mixture was stirred at 15-20°C for 2 hours. 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 obtain (1R,3S,5S)-3-((tert-butoxycarbonyl)(2-((S)-2-cyanopyrrolidine-1-yl)-2-oxoethyl)amino)adamantane-1-ylmethanesulfonate (440 g, crude) as a pale yellow oil. TLC system:PE:EA(10:1);R f :0.60) Synthesis of tert-butyl((1S,3R,5S)-3-(2-(2-azidoethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidine-1-yl)-2-oxoethyl)carbamate (Int-3) [ka]

[0149] Step 1: tert-butyl(2-((S)-2-cyanopyrrolidine-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-cyanopyrrolidine-1-yl)-2-oxoethyl)amino)adamantane-1-ylmethanesulfonate (3.58 g, 7.43 mmol) and 2,2'-oxybis(ethane-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 hours. 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 layer was dried over anhydrous sodium sulfate and evaporated to obtain the crude product. Next, the crude product was purified by silica gel column chromatography (230-400 mesh) using a 3% methanol dichloromethane solution to obtain tert-butyl(2-((S)-2-cyanopyrrolidine-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-cyanopyrrolidine-1-yl)-2-oxoethyl)amino)adamantan-1-yl)oxo)ethoxy)ethylmethanesulfonate: [ka]

[0151] To a stirred solution of tert-butyl(2-((S)-2-cyanopyrrolidine-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), triethylamine (1.67 g, 16.47 mmol) was added, 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 hour. 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 on anhydrous sodium sulfate and evaporated to obtain 2-(2-(((1R,3S,5S)-3-((tert-butoxycarbonyl)(2-((S)-2-cyanopyrrolidine-1-yl)-2-oxoethyl)amino)adamantan-1-yl)oxy)ethoxy)ethylmethanesulfonate (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-cyanopyrrolidine-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-cyanopyrrolidine-1-yl)-2-oxoethyl)amino)adamantan-1-yl)oxy)ethoxy)ethylmethanesulfonate (3.1 g, 5.44 mmol) in N,N-dimethylformamide (10 mL), sodium azide (0.71 g, 10.88 mmol) was added, and the reaction mixture was stirred at 70°C for 16 hours. 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 obtain tert-butyl((1S,3R,5S)-3-(2-(2-azidoethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidine-1-yl)-2-oxoethyl)carbamate (2.5g) 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'-(propa-2-in-1-ylazanediyl)bis(hexane-1,2,3,4,5-pentaol)(Int-4) [ka]

[0153] (2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanal (2.0 g, 11.08 mmol), propa-2-in-1-amine (0.3 g, 5.54 mmol), and acetic acid (2.0 mL) were mixed in methanol (16 mL) to which sodium cyanoborohydride (0.7 g, 11.08 mmol) was added. The reaction mixture was stirred at 60°C for 16 hours. After completion (the reaction was monitored by LC-MS), the reaction mixture was evaporated, the residue was washed with methanol (2 × 20 mL) and diethyl ether (2 × 20 mL), and dried to obtain (2R,2'R,3R,3'R,4R,4'R,5S,5'S)-6,6'-(propa-2-in-1-ylazandiyl)bis(hexane-1,2,3,4,5-pentaol) (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-cyanopyrrolidine-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-cyanopyrrolidine-1-yl)-2-oxoethyl)carbamate (1.0 g, 1.94 mmol) in methanol (10 mL), triphenylphosphine (0.76 g, 2.9 mmol) was added, and the reaction mixture was stirred at 70°C for 16 hours. After the reaction was complete, the reaction mixture was evaporated, and the residue was purified by silica gel column chromatography (basicized with triethylamine) using 6% methanol in dichloromethane to obtain tert-butyl((1S,3R,5S)-3-(2-(2-aminoethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidine-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-cyanopyrrolidine-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-cyanopyrrolidine-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 hours. After completion, the reaction mixture was evaporated to obtain a crude residue, which was then purified by silica gel (230-400 mesh) column chromatography using 3% methanol in dichloromethane to obtain tert-butyl(2-((S)-2-cyanopyrrolidine-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) Process 2 (1R,3S,5S)-3-((tert-butoxycarbonyl)(2-((S)-2-cyanopyrrolidine-1-yl)-2-oxoethyl)amino)adamantan-1-ylmethanesulfonate: [ka]

[0158] To a stirred solution of tert-butyl(2-((S)-2-cyanopyrrolidine-1-yl)-2-oxoethyl)((1S,3R,5S)-3-hydroxyadamantan-1-yl)carbamate (0.5 g, 1.24 mmol) in dichloromethane (10 mL), triethylamine (0.38 g, 3.72 mmol) was added, 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 obtain (1R,3S,5S)-3-((tert-butoxycarbonyl)(2-((S)-2-cyanopyrrolidine-1-yl)-2-oxoethyl)amino)adamantane-1-ylmethanesulfonate (0.59 g) as a pale yellow foamy solid. TLC system: MeOH:DCM (0.5:9.5);R f :0.3 (TLC eluted twice) Step 3: tert-butyl(2-((S)-2-cyanopyrrolidine-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-cyanopyrrolidine-1-yl)-2-oxoethyl)amino)adamantane-1-ylmethanesulfonate (3.58 g, 7.43 mmol) and 2,2'-oxybis(ethane-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 hours. 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 layer was dried over anhydrous sodium sulfate and evaporated to obtain the crude product. Next, the crude product was purified by silica gel column chromatography (230-400 mesh) using a 3% methanol dichloromethane solution to obtain tert-butyl(2-((S)-2-cyanopyrrolidine-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] AcOH (1 drop) was added to a solution of 4-(2-aminoethyl)phenol (15 g, 109 mmol) and hexanal (10 g, 100 mmol) in EtOH (100 mL). The reaction mixture was stirred at 25°C for 12 hours. Then, NaBH4 (2 g, 54 mmol) was added, and the reaction mixture was stirred at 25°C for 1 hour. The reaction mixture was concentrated under pressure at 40°C. The residue was diluted with water and extracted with ELISA (200 mL x 2). The organic layer was washed with brine (50 mL) and then dried on anhydrous Na2SO4. After filtration, the filtrate was concentrated under vacuum to obtain the crude product. The crude product was purified by flash column chromatography eluting with (CH2Cl2 / MeOH, 0%~10%) to obtain 4-(2-(hexylamino)ethyl)phenol (6 g, yield 24%) as a yellow oil. ES MS M / Z = 222 (M+1) Step 2: Benzylhexyl (4-hydroxyphenethyl) carbamate: [ka]

[0162] 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 to 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 hour. 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 / siRNA, 5%~20%) to obtain benzylhexyl (4-hydroxyphenethyl)carbamate (7.63 g, yield 76%) as a yellow oil. ES MS M / Z = 356 (M+1) Step 3: Benzyl (4-(2-(1,3-dioxoisoindorin-2-yl)ethoxy)phenethyl)(hexyl)carbamate: [ka]

[0163] A mixture of benzylhexyl (4-hydroxyphenethyl)carbamate (8.63 g, 24.3 mmol), 2-(2-hydroxyethyl)isoindole-1,3-dione (4.64 g, 24.3 mmol), and PPh3 (12.73 g, 48.6 mol) in THF (50 mL) was mixed with a solution of ADDP (12.24 g, 48.6 mmol) in THF (30 mL) at 25 °C under an N2 atmosphere. The reaction mixture was stirred at 70 °C for 12 hours under an N2 atmosphere. The reaction mixture was concentrated under pressure at 40 °C. The residue was purified by flash column chromatography eluting with (PE / ELISA, 10%~50%) to obtain benzyl (4-(2-(1,3-dioxoisoindorin-2-yl)ethoxy)phenethyl)(hexyl)carbamate (10.4 g, yield 81%) 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] A solution of benzyl(4-(2-(1,3-dioxoisoindoline-2-yl)ethoxy)phenethyl)(hexyl)carbamate (4.86 g, 9.20 mmol) in MeCN (10 mL) was mixed with TMSI (5 g, 35.7 mmol). The reaction mixture was stirred at 25 °C for 10 minutes. The reaction mixture was concentrated under pressure at 40 °C. The residue was purified by flash column chromatography, and elution with (CH2Cl2 / MeOH, 5%~10%) yielded 2-(2-(4-(2-(hexylamino)ethyl)phenoxy)ethyl)isoindoline-1,3-dione (2.56 g, yield 70%) 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), AcOH (10 drops) was added. The reaction mixture was stirred at 25°C for 1 hour. Then, NaBH3CN (1.86 g, 29.64 mmol) was added, and the reaction mixture was stirred at 50°C for 12 hours. (2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanal (1.33 g, 14.82 mmol) was added to this reaction mixture every 12 hours until 2-(2-(4-(2-(hexylamino)ethyl)phenoxy)ethyl)isoindoline-1,3-dione was eliminated (a total of approximately 4 equivalents of (2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanal were added to the reaction mixture, and the total reaction time was approximately 3 days). The reaction mixture was concentrated under pressure at 40°C. The residue was purified by flash column chromatography and eluted with (DCM / MeOH, 5%~10%) to obtain impure 2-(2-(4-(2-(hexyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)ethyl)phenoxy)ethyl)isoindoline-1,3-dione (4g, yield 87%) 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), hydrazine hydrate (10 mL) was added. The reaction mixture was stirred at 80 °C for 4 hours. The reaction mixture was concentrated under pressure at 40 °C. The residue was purified by flash column chromatography, and eluted with (MeOH / DCM, 20%~50%) to obtain (2R,3R,4R,5S)-6-((4-(2-aminoethoxy)phenethyl)(hexyl)amino)hexane-1,2,3,4,5-pentaol (2.5 g, yield 93%) as a colorless oil. ES MS M / Z = 429 (M+1) Synthesis of (2R,3R,4R,5S)-6-(propa-2-in-1-ylamino)hexane-1,2,3,4,5-pentaol(Int-8) [ka]

[0167] (2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanal (1.0 g, 5.54 mmol), propa-2-in-1-amine (0.61 g, 11.08 mmol), and acetic acid (1.0 mL) were mixed in methanol (20 mL), to which sodium cyanoborohydride (0.35 g, 5.54 mmol) was added at 0°C. The reaction mixture was stirred at room temperature for 16 hours. After completion (the reaction was monitored by LC-MS), the reaction mixture was evaporated, the residue was washed with diethyl ether (2 x 20 mL), and dried to obtain (2R,3R,4R,5S)-6-(propa-2-in-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]hexane-2-yl)-1-((1S,3R,5S)-3-(2-hydroxyethoxy)adamantan-1-yl)-2-oxoethyl)carbamate (Int-9) [ka]

[0169] Process 1: tert-butyl((1S)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexane-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), triethylamine (1.8 mL, 12.68 mmol), followed by di-tert-butyl dicarbonate (2.2 mL, 9.51 mmol), was added at 0°C, and the reaction mixture was stirred at room temperature for 3 hours. After completion, the reaction mixture was evaporated to obtain a crude residue, which was then purified by silica gel (230-400 mesh) column chromatography using 3% methanol in dichloromethane to obtain tert-butyl((1S)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexane-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 Process 2 (1R,3S,5S)-3-((S)-1-((tert-butoxycarbonyl)amino)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexane-2-yl)-2-oxoethyl)adamantan-1-ylmethanesulfonate: [ka]

[0171] To a stirred solution of tert-butyl((1S)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexane-2-yl)-1-((1S,3R,5S)-3-hydroxyadamantan-1-yl)-2-oxoethyl)carbamate (2.5 g, 6.02 mmol) in dichloromethane (50 mL), triethylamine (2.5 mL, 18.06 mmol) was added, 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 × 100 mL). The combined organic layers were dried over anhydrous sodium sulfate and evaporated to obtain (1R,3S,5S)-3-((S)-1-((tert-butoxycarbonyl)amino)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexane-2-yl)-2-oxoethyl)adamantane-1-ylmethanesulfonate (3.2g) crude product as a white foamy solid. TLC system: 100% HCl;R f :0.4. Step 3: tert-butyl((1S)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexane-2-yl)-1-((1S,3R,5S)-3-(2-hydroxyethoxy)adamantan-1-yl)-2-oxoethyl)carbamate(Int-9): [ka]

[0172] (1R,3S,5S)-3-((S)-1-((tert-butoxycarbonyl)amino)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexane-2-yl)-2-oxoethyl)adamantane-1-ylmethanesulfonate (3.2 g, 6.48 mmol) and ethane-1,2-diol (18.2 mL, 324.13 mmol) were stirred in acetonitrile (30 mL). Molecular sieves 4 Å (3.5 g) were added, and the resulting mixture was stirred at 70°C for 16 hours. After completion, the reaction mixture was concentrated, 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 obtain the crude product of tert-butyl((1S)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexane-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% ethyl acetate, R f :0.3 tert-butyl((1S)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexane-2-yl)-1-((1S,3R,5S)-3-(2-(2-hydroxyethoxy)ethoxy)adamantan-1-yl)-2-oxoethyl)carbamate (Int-9B) was prepared according to 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 equivalents) and potassium carbonate (594 mg, 1.5 equivalents) in methanol (8 mL), tert-butyl 4-(bromomethyl)benzylcarbamate (820 mg, 1.0 equivalent) was gradually added at 0°C. After stirring at room temperature for 4 hours, the reaction mixture was filtered using dichloromethane through a Celite pad. The filtrate was then washed with water and concentrated under vacuum to obtain a crude product as yellow oil, which was used in the next step without further purification. The crude product (700 mg) was dissolved in dioxane (4.5 mL) and methanol (0.5 mL) with 4 M HCl at 0°C. After stirring at room temperature for 3 hours, volatile substances were removed under reduced pressure to obtain a white powder of (2R,3R,4R,5S)-6-((4-(aminomethyl)benzyl)(hexyl)amino)hexane-1,2,3,4,5-pentaol hydrochloride, 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-methylbenzene sulfonate: [ka]

[0177] To a stirred solution of compound 2,2'-(ethane-1,2-diylbis(oxy))bis(ethane-1-ol) (15 g, 99.88 mmol) in DCM (150 mL), silver oxide (27.7 g, 119.85 mmol), followed by potassium iodide (1.6 g, 9.98 mmol), was added under a nitrogen atmosphere at room temperature. The resulting reaction mixture was stirred at room temperature for 10 minutes. p-TsCl (19 g, 99.88 mmol) was added, and the resulting reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by TLC. After the completion of the starting materials, 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)ethane-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), sodium azide (4.7 g, 72.28 mmol) was added at room temperature. The resulting reaction mixture was heated at 120 °C for 6 hours. The progress of the reaction was monitored by TLC. After completion of the starting materials, 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] Ph3P (3.3g, 12.56 mmol) was added to a stirred solution of CBr4 (4.1g, 12.56 mmol) in DCM (40 mL). The reaction mixture was cooled to 0°C, and compound 2-(2-(2-azidoethoxy)ethoxy)ethane-1-ol (2g, 11.42 mmol) was diluted in DCM (15 mL) and slowly added dropwise to the reaction mixture. The reaction mixture was stirred at room temperature for 8 hours. After the starting materials were complete, the reaction mixture was quenched with water (20 mL) and extracted with DCM (2 x 50 mL). The combined organic layers were 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-(buta-2-in-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-8-yl)piperidine-3-yl)carbamate (Int-13) [ka]

[0183] Step 1. 8-Bromo-7-(buta-2-in-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-bromobuta-2-yne (5.43 g, 40.81 mmol) were gradually added at room temperature. The reaction mixture was stirred at room temperature for 16 hours. After the starting materials were complete, the reaction mixture was quenched with cold water, the precipitated solid was filtered off, and dried under vacuum to obtain 9.3 g of compound 8-bromo-7-(buta-2-yne-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-(buta-2-in-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-8-yl)piperidine-3-yl)carbamate(Int-13): [ka]

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

[0187] Synthesis of methyl(R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-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), a 2 M TMS diazomethane solution in toluene (12.57 mL, 25.14 mmol) was added at 0°C, and the reaction mixture was heated to room temperature for 2 hours. After the reaction was complete, volatile substances 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% ethyl phosphate in petroleum ether used as eluent) to obtain 1.6 g of the compound methyl 6-fluoro-2-methylnicotinate as a colorless liquid product. [TLC system: ethyl phosphate: petroleum ether (3:7); R f Value: 0.8].

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

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

[0192] Step 3: Methyl(R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-6-fluoronicotinate(Int-14): [ka]

[0193] 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-(buta-2-in-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-8-yl)piperidine-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 the starting materials were completed, the reaction mixture was quenched with cold water, the precipitated solid was filtered off, and dried under vacuum to obtain 1 g of the compound methyl(R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-6-fluoronicotinate as an off-white solid. [TLC system: Depositphotos: petroleum ether (1:1); R f Value: 0.4]

[0194] Synthesis of (R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-6-(methylamino)nicotinic acid (Int-15) [ka]

[0195] Sep 1. Methyl(R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-6-(methylamino)nicotinate: [ka]

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

[0197] Step 2(R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-6-(methylamino)nicotinic acid (Int-15): [ka]

[0198] To a stirred solution of the compound methyl(R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-6-(methylamino)nicotinate (0.6 g, 1.01 mmol) in MeOH (3 mL), THF (3 mL), and water (1 mL), NaOH (0.081 g, 2.02 mmol) was added at 0°C. The reaction mixture was stirred at 50°C for 3 hours. After completion of the starting material (TLC monitoring), 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 obtain 0 g of the compound as an off-white solid. [TLC system: MeOH: DCM (1:9); R 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), NaN3 (5.04 g, 77.52 mmol) was gradually added at room temperature. The reaction mixture was stirred at 60 °C for 16 hours. After completion of the reaction (TLC monitoring), the reaction mixture was diluted with water (300 mL) and extracted with ethyl acetate (2 × 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% ethyl acetate in petroleum ether to obtain 1,7-diazidoheptane (6.5 g) as a colorless oily compound. [TLC system: ethyl acetate: 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), toluene (32.5 mL) and ethanol (32.5 mL), followed by TPP (4.68 g, 17.84 mmol), the reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction (TLC monitoring), the reaction mixture was diluted with water (200 mL) and washed with ethanol (200 mL). The aqueous layer was basicized with saturated NaHCO3 and extracted with toluene (2 × 300 mL). The combined organic layers were dried over anhydrous sodium sulfate and evaporated to obtain 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'-(propa-2-in-1-ylazandiyl)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 the compounds propa-2-in-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 hour. The reaction mixture was allowed to cool to room temperature, and then NaCNBH3 (0.68 g, 10.86 mmol) was added gradually at 0 °C. The reaction mixture was stirred at 60 °C for 16 hours. After the reaction was complete (TLC monitoring), the reaction mixture was concentrated under reduced pressure to obtain a crude mass. The crude compound was washed with Et2O (50 mL) and dried under vacuum to obtain the compound (1R,1'R,2S,2'S)-3,3'-(propa-2-in-1-ylazandiyl)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-triazole-1-yl)heptyl)amino)-2-((7-(buta-2-in-1-yl)-8-((R)-3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)nicotinate (Int-18) [ka]

[0207] Step 1: Methyl(R)-6-((7-azidoheptyl)amino)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)nicotinate: [ka]

[0208] Compound 3 (1.41 g, 9.00 mmol) and K2CO3 (1.24 g, 9.00 mmol) were added to a solution of compound methyl(R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-6-fluoronicotinate (3.5 g, 6.00 mmol) in DMF (30 mL), and the reaction mixture was stirred at 80 °C for 16 hours. After the reaction was complete (TLC monitoring), the reaction mixture was diluted with water (100 mL) and extracted with ELISA (2 × 100 mL). The combined organic layer was dried over anhydrous sodium sulfate and evaporated to obtain the crude product. The crude product obtained was purified by silica gel (100-200 mesh) column chromatography using 30% SiO in petroleum ether to obtain methyl(R)-6-((7-azidoheptyl)amino)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)nicotinate (2.8 g) as an off-white solid. [TLC system: SiO: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-triazole-1-yl)heptyl)amino)-2-((7-(buta-2-in-1-yl)-8-((R)-3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)nicotinate (Int-18): [ka]

[0210] The compounds methyl(R)-6-((7-azidoheptyl)amino)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)nicotinate (0.5g, 0.69 mmol) and (1R,1'R,2S,2'S)-3,3'-(propa-2-in-1-ylazandiyl)bis(1-((4R,5R)-5-hydroxy-2-phenyl-1,3-dioxan-4-yl)propane-1,2-diol) (0.97g) To a solution of 3.62 mmol) of 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 hours. After the reaction was complete (TLC monitoring), the reaction mixture was concentrated under reduced pressure to obtain a crude mass. The crude compound was washed with Et2O (50 mL) and dried under vacuum to obtain the 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-triazole-1-yl)heptyl)amino)-2-((7-(buta-2-in-1-yl)-8-((R)-3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)nicotinate (0.9 g) as an off-white solid. The crude compound was carried 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] To a stirred solution of methyl 5-chloro-2-methylbenzoate (10.0 g, 54.16 mmol) in 1,2-dichloroethane (100 mL), N-bromosuccinimide (9.64 g, 54.16 mmol) and catalyst benzoyl peroxide (0.66 g, 2.71 mmol) were gradually added. The reaction mixture was stirred at 80°C for 5 hours. After the starting materials were removed, the reaction mixture was evaporated under reduced pressure, quenched with water, and extracted with ethyl acetate (3 × 100 mL). The organic layer was washed with brine solution (150 mL), dried on 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 8 g of the compound methyl 2-(bromomethyl)-5-chlorobenzoate as a colorless liquid. [TLC system: petroleum ether; R f Value: 0.7]

[0213] Synthesis of (R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-5-chlorobenzoic acid (Int-20) [ka]

[0214] Step 1. 8-Bromo-7-(buta-2-in-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-bromobuta-2-yne (5.43 g, 40.81 mmol) were gradually added at room temperature. The reaction mixture was stirred at room temperature for 16 hours. After the starting materials were complete, the reaction mixture was quenched with cold water, the precipitated solid was filtered off, and dried under vacuum to obtain 9.3 g of compound 8-bromo-7-(buta-2-yne-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-(buta-2-in-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-5-chlorobenzoate: [ka]

[0217] To a stirred solution of compound 8-bromo-7-(buta-2-in-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 the starting materials were completed, the reaction mixture was quenched with cold water, the precipitated solid was filtered off, and dried under vacuum to obtain 2.5 g of compound methyl 2-((8-bromo-7-(buta-2-in-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: SiO: Petroleum Ether (1:1); R f Value: 0.7]

[0218] Step 3: Ethylmethyl(R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-5-chlorobenzoate: [ka]

[0219] To a stirred solution of compound methyl 2-((8-bromo-7-(buta-2-in-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-5-chlorobenzoate (2.5 g, 5.21 mmol) in DMF (25 mL), compound tert-butyl(R)-piperidine-3-ylcarbamate (1.25 g, 6.25 mmol) and K2CO3 (2.16 g, 15.63 mmol) were added, and the reaction mixture was stirred at 65°C for 8 hours. After the starting materials were completed, the reaction mixture was quenched with cold water, the precipitated solid was filtered off, and dried under vacuum to obtain 2.0 g of the compound ethylmethyl(R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-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: ԅ:petroleum ether (1:1);R f Value: 0.3].

[0220] Step 4(R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-5-chlorobenzoic acid (Int-20): [ka]

[0221] To a stirred solution of the compound ethylmethyl(R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-5-chlorobenzoate (0.5 g, 0.83 mmol) in MeOH (5 mL) and water (1 mL), LiOH.H2O (0.11 g, 2.5 mmol) was added at 0°C. The reaction mixture was stirred at room temperature for 16 hours. After the starting material was removed, 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 obtain 0.4 g of compound (R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-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), 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) were added. The reaction mixture was stirred at room temperature for 16 hours, and the progress of the reaction was monitored by TLC. After the completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain the crude compound. The crude compound was purified by silica gel (Davisil) column chromatography (using 0-60% ethyl ether in petroleum ether as an eluent) to obtain 1.6 g of the compound methyl(R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate as a white solid. [TLC system: ethyl ether: petroleum ether (6:4); R f Value: 0.5]

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

[0226] To a solution of the compound methyl(R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5trifluorophenyl)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), a 4 M NaOH solution (2.7 mL, 5.5 V) was added at 0°C. The reaction mixture was then stirred at room temperature for 2 hours. After the reaction was complete, the reaction mixture was concentrated, the residue was acidified to pH-4 with 10% HCl, and then the reaction mixture was concentrated again to obtain the residue, which was dried by co-evaporation with ACN and toluene to obtain 0.450 g of compound (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5trifluorophenyl)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-(2,4,5trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (Int-22) [ka]

[0228] 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) was stirred in DCM (10 mL) to which Et3N (0.341 mL, 2.45 mmol) and BOP-Cl (0.312 g, 1.22 mmol), followed by compound 2-(2-(2-azidoethoxy)ethoxy)ethane-1-ol (0.214 g, 1.22 mmol). The reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by TLC. After the reaction was complete, the reaction mixture was concentrated to obtain the crude compound, which was purified by silica gel (Davisil) column chromatography (using 0-60% ethyl ether in petroleum ether as the 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: ethyl:petroleum ether (7:3); R f Value: 0.7]

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

[0230] 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) was stirred 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 the starting material was complete, 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: siRNA: petroleum ether (7:3); R f Value: 0.2].

[0231] Exemplary Compounds in this Disclosure [Examples] [Example 1] Synthesis of heptyl(3-((2-((S)-2-cyanopyrrolidine-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 to DCM [15 mL]. The reaction mixture was stirred under N2 at room temperature for 2 hours. The mixture was filtered and washed three times with DCM (5 mL). The filtrate was concentrated to obtain methyl 6-chloro-4-methoxypyridazine-3-carboxylate (270 mg, yield 14%) as a white solid. ES MS M / Z = 309 (M + 1) Step 2: Heptyl(3-((2-((S)-2-cyanopyrrolidine-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 to MeCN [10 mL]. The reaction mixture was stirred at room temperature for 2 hours. The mixture was concentrated, and the residue was purified by silica gel column chromatography eluted with (DCM:MeOH = 20:1) to obtain heptyl (3-((2-((S)-2-cyanopyrrolidine-1-yl)-2-oxoethyl)amino)adamantan-1-yl)carbamate (1,160 mg, yield 90%) 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), a solution of CbzCl (5.08 g, 29.9 mmol) in DCM (20 mL) was added at 0°C. The reaction mixture was stirred at 25°C for 1 hour. 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 and eluted with (MeOH / DCM, 5%~10%) to obtain benzyl(3-hydroxyadamantan-1-yl)carbamate (8.5 g, yield 94%) as a white solid. ES MS M / Z = 302 (M+1) Step 2.3-(((benzyloxy)carbonyl)amino)adamantan-1-ylmethanesulfonate: [ka]

[0237] 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 at 0°C to a solution of methanesulfonyl chloride (1.71 g, 15 mmol) in DCM (2 mL). The reaction mixture was stirred at 0°C for 1 hour. 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 obtain 3-(((benzyloxy)carbonyl)amino)adamantan-1-ylmethanesulfonate (3.3 g, yield 87%) as a white solid, which was used in the next step without further purification. ES MS M / Z = 402 (M + 23) Process 3S-(3-aminoadamantan-1-yl)ethanethioate: [ka]

[0238] A solution of 3-(((benzyloxy)carbonyl)amino)adamantane-1-ylmethanesulfonate (2.5 g, 6.59 mmol) in thiol acetate (15 mL) was stirred at 100 °C for 12 hours. The reaction mixture was concentrated under pressure at 45 °C. The residue was purified by flash column chromatography, and eluted with (MeOH / DCM, 5%~10%) to obtain S-(3-aminoadamantane-1-yl)ethanethioate (1 g, yield 67%) as a yellow solid. ES MS M / Z = 226 (M+1) Process 4S-(3-((2-((S)-2-cyanopyrrolidine-1-yl)-2-oxoethyl)amino)adamantan-1-yl)ethanethioate: [ka]

[0239] 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 mixed with 2-chloro-1-(pyrrolidine-1-yl)ethane-1-one (58 mg, 0.4 mmol). The reaction mixture was stirred at 75°C for 6 hours. The reaction mixture was concentrated under pressure at 40°C. The residue was purified by flash column chromatography, and eluted with MeOH / DCM (5%~10%) to obtain S-(3-((2-((S)-2-cyanopyrrolidine-1-yl)-2-oxoethyl)amino)adamantan-1-yl)ethanethioate (128 mg, yield 71%) 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-cyanopyrrolidine-1-yl)-2-oxoethyl)amino)adamantan-1-yl)ethanethioate (1 g, 2.77 mmol) in MeOH / H2O (V / V=20:1, 10 mL), K2CO3 (1.14 g, 8.31 mmol) was added. The reaction mixture was stirred at 25°C for 30 minutes. The reaction mixture was dried over Na2SO4 and concentrated under pressure at 35°C. The residue was purified by flash column chromatography and eluted with (MeOH / DCM, 10%~20%) to obtain (2S)-1-((3-mercaptoadamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (300 mg, yield 33%) as a yellow oil. ES MS M / Z=320(M+1) Step 6: tert-butyl(2-((S)-2-cyanopyrrolidine-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 under reflux for 12 hours. The mixture was concentrated. The residue was purified by flash column chromatography, and eluted with MeOH / DCM (5%~10%) to obtain tert-butyl(2-((S)-2-cyanopyrrolidine-1-yl)-2-oxoethyl)(3-mercaptoadamantan-1-yl)carbamate (400 mg, yield 76%) as a colorless oil. ES MS M / Z = 442 (M + 23) Step 7: tert-butyl(2-((S)-2-cyanopyrrolidine-1-yl)-2-oxoethyl)(3-(heptylthio)adamantan-1-yl)carbamate: [ka]

[0242] A mixture of tert-butyl(2-((S)-2-cyanopyrrolidine-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 hours. The mixture was concentrated, and the residue was purified by silica gel column chromatography eluted with (PE:siRNA=2:1) ​​to obtain tert-butyl(2-((S)-2-cyanopyrrolidine-1-yl)-2-oxoethyl)(3-(heptylthio)adamantan-1-yl)carbamate (50 mg, yield 27%) as a yellow solid. ES MS M / Z=540(M+23) Process 8(2S)-1-((3-(heptylthio)adamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile(2): [ka]

[0243] A solution of tert-butyl(2-((S)-2-cyanopyrrolidine-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 minutes. The mixture was concentrated, and the residue was purified by preparative HPLC (Gemini-C18 150 × 21.2 mm, 5 μm, mobile phase: ACN-H2O, gradient: 30-70%) to obtain (2S)-1-((3-(heptylthio)adamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (32 mg, yield 80%) 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-carbonitrili (3) [ka]

[0244] To a solution of tert-butyl(2-((S)-2-cyanopyrrolidine-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(heptylthio)adamantan-1-yl)carbamate (20 mg, 0.03 mmol) in DCM (3 mL) under a nitrogen atmosphere, 3-chloroperbenzoic acid (10 mg, 0.05 mmol) was added dropwise at -40°C. The reaction mixture was stirred at -40°C for 30 minutes. TFA (1 mL) was slowly added. The reaction mixture was stirred at room temperature for a further 30 minutes. The reaction mixture was concentrated under pressure. Next, the crude product was purified by general preparative HPLC (Gemini-C18 150×21.2mm, 5um, mobile phase: ACN-H2O, 0.1% FA, gradient: 5~50%) to obtain 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-cyanopyrrolidine-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-cyanopyrrolidine-1-yl)-2-oxoethyl)(3-((2-hydroxyethyl)thio)adamantan-1-yl)carbamate A mixture of tert-butyl(2-((S)-2-cyanopyrrolidine-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 mixed with 2-iodoethanol (1.42 g, 8.3 mmol). The reaction mixture was stirred at 25°C for 12 hours. The reaction mixture was concentrated under pressure. The residue was purified by flash chromatography and eluted with CH2Cl2 / MeOH (10:1) to obtain the product tert-butyl(2-((S)-2-cyanopyrrolidine-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-cyanopyrrolidine-1-yl)-2-oxoethyl)(3-((2-(((4-nitrophenoxy)carbonyl)oxy)ethyl)thio)adamantan-1-yl)carbamate A mixture of tert-butyl(2-((S)-2-cyanopyrrolidine-1-yl)-2-oxoethyl)(3-((2-hydroxyethyl)thio)adamantan-1-yl)carbamate (340 mg, 0.73 mmol) and Et3N (221 mg, 2.19 mmol) in DCM (20 mL) was mixed with 4-nitrophenyl chloroformate (220 mg, 1.09 mmol). The reaction mixture was stirred at 25°C for 3 hours. H2O (20 mL) was added. The residue was extracted with DCM (30 mL). When the DCM phase was concentrated, tert-butyl(2-((S)-2-cyanopyrrolidine-1-yl)-2-oxoethyl)(3-((2-(((4-nitrophenoxy)carbonyl)oxy)ethyl)thio)adamantan-1-yl)carbamate (600 mg, crude) was obtained 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-cyanopyrrolidine-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)carbamate)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), TEA (303 mg, 3 mmol) was added, followed by a solution of tert-butyl(2-((S)-2-cyanopyrrolidine-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 hours. The reaction mixture was concentrated under pressure. The residue was purified by flash chromatography and eluted with CH2Cl2 / MeOH (5:1) to obtain the product tert-butyl(2-((S)-2-cyanopyrrolidine-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-cyanopyrrolidine-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 400 mg, 0.43 mmol) of tert-butyl(2-((S)-2-cyanopyrrolidine-1-yl)-2-oxoethyl)((1S,3R,5S)-3-((2-(((2-((2-(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) was dissolved in 15 mL of DCM, to which ZnBr2 (1086 mg, 4.8 mmol) was added. The reaction mixture was stirred at 25°C for 8 hours. The solution was filtered, the filtrate was concentrated, and purified by preparative TLC (CH2Cl2 / MeOH) = (5:1) to obtain impurities. The impurities were then purified by general preparative HPLC (Gemini-C18 150×21.2mm, 5um, mobile phase: ACN-H2O, 0.1% FA, gradient: 5~50%) to obtain product 2-(((1R,3S,5S)-3-((2-((S)-2-cyanopyrrolidine-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-carbonitrili (5) [ka]

[0246] Step 1: tert-butyl(2-((S)-2-cyanopyrrolidine-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-cyanopyrrolidine-1-yl)-2-oxoethyl)amino)adamantane-1-ylmethanesulfonate (Int-2, 0.59 g, 1.22 mmol) and ethane-1,2-diol (3.8 g, 61.25 mmol) in acetonitrile (3.0 mL), a molecular sieve 4 Å (2.0 g) was added, and the resulting mixture was stirred at 70°C for 16 hours. After completion, the reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (3 × 50 mL). The combined organic layer was dried over anhydrous sodium sulfate and evaporated to obtain the crude product. Next, the crude product was purified by silica gel column chromatography using 3% methanol in dichloromethane, and further purified by RP-HPLC under the following conditions: Column / dimensions: X-BRIDGE-C 18 (19*250, 5um), Mobile phase A: 10mM ammonium bicarbonate in water, Mobile phase B: ACN. Gradient (time / B%): 00 / 10, 3 / 10, 7 / 35, 20 / 75, 20.1 / 100, 22 / 100, 22.1 / 10, 24 / 10, Flow rate: 17 ml / min. Upon evaporation of the desired fraction, tert-butyl(2-((S)-2-cyanopyrrolidine-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-hydroxyethoxy)adamantan-1-yl)carbamate (0.1 g) was obtained as an off-white solid. TLC system: MeOH:DCM (1:9); R 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-cyanopyrrolidine-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-hydroxyethoxy)adamantan-1-yl)carbamate (0.09 g, 0.2 mmol) in acetonitrile (5 mL), a 1 M tin(IV) chloride solution in heptane (0.8 mL, 0.8 mmol) was added, and the mixture was stirred at room temperature for 3 hours. After completion, the reaction mixture was evaporated, the residue was co-evaporated with methanol (10 mL), and washed with diethyl ether (2 x 5 mL) to obtain 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 fraction was freeze-dried to obtain (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);R 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-cyanopyrrolidine-1-yl)-2-oxoethyl)amino)adamantan-1-yl)oxy)methyloctanoate (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 overnight at room temperature. The reaction mixture was diluted with siRNA (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 silica gel column chromatography eluted with (PE / siRNA=8 / 1) to obtain benzyl(3-((methylthio)methoxy)adamantan-1-yl)carbamate (2.6 g, yield 72.2%) as a pale yellow oil. ES MS M / Z=384(M+23) Step 2. ((3-(((benzyloxy)carbonyl)amino)adamantan-1-yl)oxy)methyloctanoate: [ka]

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

[0252] To a solution of -(((benzyloxy)carbonyl)amino)adamantan-1-yl)oxy)methyloctanoate (1.7 g, 3.72 mmol) in MeOH (10 mL), Pd / C (500 mg) was added. The reaction mixture was stirred overnight at room temperature under an H2 atmosphere. The mixture was filtered, and the filtrate was evaporated under reduced pressure to obtain ((3-aminoadamantan-1-yl)oxy)methyloctanoate (1.1 g, yield 91.4%) as a colorless oil. ES MS M / Z = 324(M+1) Step 4. ((3-((2-((S)-2-cyanopyrrolidine-1-yl)-2-oxoethyl)amino)adamantan-1-yl)oxy)methyloctanoate (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 hours under an N2 atmosphere. The mixture was diluted with RINKAN (30 mL) and washed with water (10 mL x 2). The organic phase was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. Purification of the crude product by silica gel column chromatography using (PE / siRNA=6 / 1) elution yielded ((3-((2-((S)-2-cyanopyrrolidine-1-yl)-2-oxoethyl)amino)adamantan-1-yl)oxy)methyloctanoate (mCMP694) (400 mg, yield 28%) 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-triazole-1-yl)ethoxy)ethoxy)ethoxy)adamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (8): [ka]

[0254] Step 1: tert-butyl(2-((S)-2-cyanopyrrolidine-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-cyanopyrrolidine-1-yl)-2-oxoethyl)amino)adamantane-1-ylmethanesulfonate (137 g, 285 mmol, 1.00 equivalent) in THF (1.40 L), TEA (66.1 g, 653 mmol, 2.29 equivalents) and 2,2'-(ethane-1,2-diylbis(oxy))bis(ethane-1-ol) (722 g, 4.81 mol, 16.8 equivalents) were added. The reaction mixture was heated at 70°C for 48 hours. The reaction mixture was diluted with H2O (2.00 L) and extracted with EA (2.00 L x 2). The combined organic layers were washed with 2.00 L of 5% citric acid aqueous solution, then washed with 2.00 L of saturated NaHCO3 aqueous solution, 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 obtain the compound tert-butyl(2-((S)-2-cyanopyrrolidine-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-cyanopyrrolidine-1-yl)-2-oxoethyl)amino)adamantan-1-yl)oxo)ethoxy)ethoxy)ethylmethanesulfonate: [ka]

[0257] To a solution of the compound tert-butyl(2-((S)-2-cyanopyrrolidine-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)adamantan-1-yl)carbamate (162 g, 302 mmol, 1.00 equivalent) and N,N,N',N'-tetramethyl-1,6-hexanediamine (115 g, 667 mmol, 2.21 equivalents) in DCM (1600 mL), MsCl (54.9 g, 479 mmol, 37.1 mL, 1.59 equivalents) was added at 0-10°C. The reaction mixture was stirred at 0-10°C for 2 hours. The reaction mixture was quenched at 0°C by adding ice water (1.00 L), then titrated with 5% citric acid aqueous solution (2.00 L) until the pH reached 5-6, and extracted with EA (3.00 L). The combined organic layer was washed with NaHCO3 (1.00 L), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain compound 2-(2-(2-(((1R,3S,5S)-3-((tert-butoxycarbonyl)(2-((S)-2-cyanopyrrolidine-1-yl)-2-oxoethyl)amino)adamantan-1-yl)oxy)ethoxy)ethoxy)ethylmethanesulfonate (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-cyanopyrrolidine-1-yl)-2-oxoethyl)carbamate: [ka]

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

[0260] Process 4 tert-butyl(2-((S)-2-cyanopyrrolidine-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-triazole-1-yl)ethoxy)ethoxy)ethoxy)adamantan-1-yl)carbamate: [ka]

[0261] A mixture of the compounds tert-butyl((1S,3R,5S)-3-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidine-1-yl)-2-oxoethyl)carbamate (152 g, 271 mmol, 1.00 equivalent), (2R,3R,4R,5S)-6-(hexyl(propa-2-in-1-yl)amino)hexane-1,2,3,4,5-pentaol (98.8 g, 325 mmol, 1.20 equivalent), and CuI (6.84 g, 35.9 mmol, 0.13 equivalent) in THF (1.52 L) was stirred at 25°C for 12 hours. The reaction mixture was diluted with H2O (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 obtain the compound tert-butyl(2-((S)-2-cyanopyrrolidine-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-triazole-1-yl)ethoxy)ethoxy)ethoxy)adamantan-1-yl)carbamate (100 g, 115 mmol, yield 42.7%, purity 41.3%) 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-triazole-1-yl)ethoxy)ethoxy)ethoxy)adamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile(8): [ka]

[0262] To a solution of the compound tert-butyl(2-((S)-2-cyanopyrrolidine-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-triazole-1-yl)ethoxy)ethoxy)ethoxy)adamantan-1-yl)carbamate (40.0 g, 46.3 mmol, 1.00 equivalent) was added at 0°C. The reaction mixture was stirred at 25°C for 18 hours. The reaction mixture was extracted with DCM (1.00 L x 2). The aqueous phase was adjusted to pH 9-10 using saturated NaHCO3 and K2CO3, and extracted using DCM (1.00 L x 2). The combined organic phase was dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain 13 (24.5 g, 31.0 mmol, yield 66.9%, purity 96.7%) 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 pathway 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-triazole-1-yl)ethoxy)ethoxy)adamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (9): [ka]

[0264] Process 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-triazole-1-yl)ethoxy)ethoxy)adamantan-1-yl)(2((S)-2-cyanopyrrolidine-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-cyanopyrrolidine-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'-(propa-2-in-1-ylazandiyl)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 LC-MS), the reaction mixture was filtered through Celite, washed with methanol and water (1:1) (100 mL), and the filtrate was evaporated to obtain 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. The fraction containing the desired product was recovered and evaporated to obtain 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-triazole-1-yl)ethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidine-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-triazole-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-triazole-1-yl)ethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidine-1-yl)-2-oxoethyl)carbamate (0.35 g, 0.39 mmol) in acetonitrile (15 mL), a 1 M tin(IV) chloride solution in heptane (1.55 mL, 1.55 mmol) was added, and the reaction mixture was stirred at room temperature for 3 hours. After completion (the reaction was monitored by LC-MS), the reaction mixture was evaporated, the residue was co-evaporated with methanol (10 mL), and the mixture was washed with diethyl ether (2 × 5 mL) to obtain the crude product. Next, the crude product was purified by C-18 column chromatography using 30% methanol in 10 mM ammonium bicarbonate in water. The desired fraction was lyophilized to obtain (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-triazole-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] Process 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-cyanopyrrolidine-1-yl)-2-oxoethyl)carbamate: [ka]

[0269] Sodium cyanoborohydride (0.034 g, 0.52 mmol) was added to a mixture of tert-butyl((1S,3R,5S)-3-(2-(2-aminoethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidine-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). The reaction mixture was stirred at 70 °C for 24 hours. After the reaction was complete (monitored by LC-MS), the reaction mixture was evaporated to obtain the crude product. The crude product was purified by reverse-phase C18 column chromatography using 50% methanol in 0.1% formic acid in water. The fraction containing the desired product was recovered and evaporated to obtain 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-cyanopyrrolidine-1-yl)-2-oxoethyl)carbamate (0.28 g) as a colorless 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-cyanopyrrolidine-1-yl)-2-oxoethyl)carbamate (0.22 g, 0.27 mmol) in water (5 mL), a 2N hydrochloric acid aqueous solution was added, and the reaction mixture was stirred at room temperature for 24 hours. After the reaction was complete (monitored by LC-MS), the reaction mixture was purified by RP preparative HPLC using the following conditions: Column / Dimensions: X-BRIDGE (19*150), 5um, Mobile Phase A: 10mM ammonium bicarbonate (aqueous solution) in water, Mobile Phase B: ACN, Gradient (time / B%): 0 / 2, 3 / 2, 16 / 36, 16.1 / 100, 18 / 100, 18.1 / 2, 20 / 2, Flow Rate: 17 ml / min. The desired fraction was freeze-dried to obtain (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-cyanopyrrolidine-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] Sodium cyanoborohydride (0.03 g, 0.45 mmol) was added to a mixture of tert-butyl((1S,3R,5S)-3-(2-(2-aminoethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidine-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). The reaction mixture was stirred at 60 °C for 24 hours. After completion (the reaction was monitored by LC-MS), the mixture was evaporated under reduced pressure to obtain the crude product. Next, the crude product was purified by reverse-phase preparative HPLC using the following conditions: Column / dimensions: X-BRIDGE (19*150), 5um, Mobile phase A: 10mM ammonium bicarbonate (aqueous solution) in water, Mobile phase B: ACN, Gradient (time / B%): 0 / 10, 1 / 10, 20 / 50, 20 / 10 / 95, 22 / 10 / 95, 22 / 20 / 10, 24 / 10, Flow rate: 17 ml / min. The desired fraction was recovered and freeze-dried to obtain tert-butyl(2-((S)-2-cyanopyrrolidine-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] Trifluoroacetic acid was added at 0°C to a stirred suspension of tert-butyl(2-((S)-2-cyanopyrrolidine-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), and the reaction mixture was stirred at room temperature for 1 hour. After completion (monitored by LC-MS), the reaction mixture was evaporated under reduced pressure to obtain the crude product. Next, the crude product was purified by reverse-phase preparative HPLC using the following conditions: Column / dimensions: X-BRIDGE (19*150), 5um, Mobile phase A: 10mM ammonium bicarbonate (aqueous solution) in water, Mobile phase B: ACN, Gradient (time / B%): 0 / 10, 1 / 10, 11 / 45 / 20, 11 / 10 / 95, 13 / 10 / 95, 13 / 20 / 10, 15 / 10, Flow rate: 17 ml / min. When the desired fraction was freeze-dried, (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%) was obtained 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-cyanopyrrolidine-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-pentaol: [ka]

[0277] 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 mixed with Raney nickel (5 g), and the reaction mixture was stirred at 70°C for 16 hours under a hydrogen gas pressure of 150 psi. After completion (monitored by LC-MS), the reaction mixture was filtered through a Celite pad and washed with methanol (50 mL). The filtrate was evaporated to obtain a solid, which was ground with diethyl ether (50 mL) and dried to obtain (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-pentaol (1.0 g, 4.78 mmol) in methanol (10 mL), potassium carbonate (0.99 g, 7.17 mmol), followed by tert-butyl (4-(bromomethyl)benzyl)carbamate (1.44 g, 4.78 mmol), was gradually added at 0°C, and the mixture was stirred at room temperature for 16 hours. After completion (monitored by LC-MS), the reaction mixture was evaporated to obtain the crude product. The crude product was then purified by reverse-phase C18 flash column chromatography using 18% aqueous solution of 0.1% formic acid and methanol as the mobile phase. The fraction containing the desired product was recovered and evaporated to obtain 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-pentaol 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), 4 M hydrochloric acid in 1,4-dioxane was slowly added at 0°C, and the resulting mixture was stirred at room temperature for 2 hours. After completion (monitored by LC-MS), the reaction mixture was evaporated, yielding (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-cyanopyrrolidine-1-yl)-2-oxoethyl)amino)adamantan-1-yl)oxo)ethoxy)ethyl(4-nitrophenyl)carbonate: [ka]

[0283] (2S)-1-(((1S,3R,5S)-3-(2-(2-hydroxyethoxy)ethoxy)adamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile (0.2 g, 0.407 mmol) was stirred in dichloromethane (4 mL), to which triethylamine (0.164 g, 1.628 mmol) was added, followed by 4-nitrophenylcarbonochloride (0.09 g, 0.447 mmol) in dichloromethane (1 mL) at -10°C. The mixture was stirred at room temperature for 2 hours. After completion, the reaction mixture was directly incorporated into the next step.

[0284] Step 5.2-(2-(((1R,3S,5S)-3-((2-((S)-2-cyanopyrrolidine-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), triethylamine (0.24 g, 2.34 mmol) was added, followed by 2-(2-(((1R,3S,5S)-3-((2-((S)-2-cyanopyrrolidine-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 hours. After completion (monitoring by LC-MS), the reaction mixture was evaporated to obtain the crude product, which was then purified by reverse-phase C18 column chromatography using a 40% aqueous solution of 0.1% formic acid and acetonitrile as the mobile phase. The fraction containing the desired product was recovered and evaporated to obtain 2-(2-(((1R,3S,5S)-3-((2-((S)-2-cyanopyrrolidine-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-cyanopyrrolidine-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] Trifluoroacetic acid was added at 0°C to a stirred suspension of 2-(2-(((1R,3S,5S)-3-((2-((S)-2-cyanopyrrolidine-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), and the reaction mixture was stirred at room temperature for 3 hours. After completion (monitored by LC-MS), the reaction mixture was concentrated, the residue was washed with diethyl ether (3 x 10 mL), and dried to obtain the crude product. Next, the crude product was purified by reversed-phase preparative HPLC using the following conditions: Column / Dimensions: SUNFIREC18 (19*150*5μ), Mobile Phase A: 0.1% FA (aqueous solution) in water, Mobile Phase B: Acetonitrile, Gradient (Time / B%): 0 / 5, 2 / 5, 10 / 20, 10.1 / 100, 14 / 100, 14.1 / 5, 16 / 5, Flow Rate: 17 ml / min. When the desired fraction was freeze-dried, 2-(2-(((1R,3S,5S)-3-((2-((S)-2-cyanopyrrolidine-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) was obtained 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-cyanopyrrolidine-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-cyanopyrrolidine-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-medium cyanopyrrolidine-1-yl)-2-oxoethyl)amino)adamantane-1-ylmethanesulfonate (1.25 g, 2.59 mmol) in MeCN (5 mL), ethane-1,2-diol (8.04 g, 129.6 mmol) was added. The reaction mixture was stirred at 70°C for 12 hours. The reaction mixture was concentrated under pressure at 40°C. The residue was purified by flash column chromatography and eluted with (MeOH / DCM, 1%~10%) to obtain tert-butyl(2-((S)-2-cyanopyrrolidine-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-hydroxyethoxy)adamantan-1-yl)carbamate (860 mg, yield 74%) as a yellow oil. ES MS M / Z = 470 (M + 23) Step 2 tert-butyl(2-((S)-2-cyanopyrrolidine-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-cyanopyrrolidine-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), 4-nitrophenyl chloroformate (800 mg, 4 mmol) was added. The reaction mixture was stirred at 25°C for 3 hours. The mixture was quenched with H2O (20 mL) and extracted with DCM (30 mL). The organic layer was dried over Na2SO4 and heated under pressure for 30°C. o When concentrated with C, tert-butyl(2-((S)-2-cyanopyrrolidine-1-yl)-2-oxoethyl)((1S,3R,5S)-3-(2-(((4-nitrophenoxy)carbonyl)oxy)ethoxy)adamantan-1-yl)carbamate (1.2g, crude) was obtained 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-cyanopyrrolidine-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), TEA (210 mg, 2.1 mmol) was added, followed by a solution of tert-butyl(2-((S)-2-cyanopyrrolidine-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 hours. The reaction mixture was concentrated under pressure at 40°C. The residue was purified by flash chromatography and eluted with (MeOH / DCM, 10%~50%) to obtain tert-butyl(2-((S)-2-cyanopyrrolidine-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, yield 69%) as a yellow oil. ES MS M / Z = 902 (M+1) Step 3.2-(((1R,3S,5S)-3-((2-((S)-2-cyanopyrrolidine-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] 436 mg, 0.48 mmol) of tert-butyl(2-((S)-2-cyanopyrrolidine-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) was dissolved in 15 mL of DCM, to which ZnBr2 (1086 mg, 4.8 mmol) was added. The reaction mixture was stirred at 25°C for 12 hours. 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 obtain impurities, which were then purified by general preparative HPLC (Gemini-C18 150×21.2mm, 5um, mobile phase: ACN-H2O, 0.1% FA, gradient: 5~50%) to obtain 2-(((1R,3S,5S)-3-((2-((S)-2-cyanopyrrolidine-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: A mixture of tert-butyl(2-((S)-2-cyanopyrrolidine-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-triazole-1-yl)ethoxy)ethoxy)adamantan-1-yl)carbamate and tert-butyl((1S,3R,5S)-3-(2-(2-(4-(aminomethyl)-1H-1,2,3-triazole-1-yl)ethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidine-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-cyanopyrrolidine-1-yl)-2-oxoethyl)carbamate (1.0 g, 1.94 mmol), (2R,3R,4R,5S)-6-(propa-2-in-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 obtain 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. The fraction containing the desired product was recovered and evaporated to obtain tert-butyl(2-((S)-2-cyanopyrrolidine-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-triazole-1-yl)ethoxy)ethoxy A mixture of (adamantan-1-yl) carbamate and tert-butyl ((1S,3R,5S)-3-(2-(2-(4-(aminomethyl)-1H-1,2,3triazole-1-yl)ethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidine-1-yl)-2-oxoethyl) carbamate (0.59 g, crude) was obtained as a brown solid.

[0295] Step 2: tert-butyl(2-((S)-2-cyanopyrrolidine-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-triazole-1-yl)ethoxy)ethoxy)adamantan-1-yl)carbamate: [ka]

[0296] A mixture of tert-butyl((1S,3R,5S)-3-(2-(2-(4-(aminomethyl)-1H-1,2,3-triazole-1-yl)ethoxy)ethoxy)adamantan-1-yl)(2-((S)-2-cyanopyrrolidine-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) was mixed with sodium cyanoborohydride (0.048 g, 0.77 mmol), and the mixture was stirred at 60°C for 6 hours. After completion, the reaction mixture was evaporated to obtain the crude product, which was then purified by RP preparative HPLC under the following conditions. Column / Dimensions: X-BRIDGEC18 (19*250) 5um, Mobile Phase A: 10mM ABC (aqueous solution) in water, Mobile Phase B: 100% ACN, Gradient (Time / B %): 0 / 20, 3 / 20, 18 / 40, 18.1 / 100, 20 / 100, 20.1 / 20, 24 / 20, Flow Rate: 17 ml / min. When the fraction containing the desired product was evaporated, tert-butyl(2-((S)-2-cyanopyrrolidine-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-triazole-1-yl)ethoxy)ethoxy)adamantan-1-yl)carbamate (0.1g) was obtained 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-triazole-1-yl)ethoxy)ethoxy)adamantan-1-yl)glycyl)pyrrolidine-2-carbonitrile(14): [ka]

[0298] To a stirred solution of tert-butyl(2-((S)-2-cyanopyrrolidine-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-triazole-1-yl)ethoxy)ethoxy)adamantan-1-yl)carbamate (0.1 g, 0.122 mmol) in a mixture of acetonitrile and water (4 mL), 1.22 mL, 2.44 mmol of 2N hydrochloric acid aqueous solution was added, and the reaction mixture was stirred at room temperature for 24 hours. After the completion of the reaction (monitored by LC-MS), the reaction mixture was evaporated to obtain the crude product. The obtained crude product was purified by RP preparative HPLC under the following conditions. Column / dimensions: X BRIDGE C18 (19 × 250 mm), 5 μm, mobile phase A: 10 mM ABC in water, mobile phase B: acetonitrile, gradient (time / B%): 0 / 5, 4 / 5, 12 / 75, 12.1 / 5, 15 / 5, flow rate: 16 ml / min. Freeze-drying of the desired peak fraction yielded (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-triazole-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-carbonitrili (16): [ka]

[0299] To a stirred solution of tert-butyl((1S)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexane-2-yl)-1-((1S,3R,5S)-3-(2-hydroxyethoxy)adamantan-1-yl)-2-oxoethyl)carbamate (0.30 g, 0.65 mmol), DCM (3 mL) and TFA (3 mL) were added dropwise at 0°C, and the reaction mixture was stirred at room temperature for 3 hours. After the completion of the reaction (monitored by LC-MS), the reaction mixture was concentrated, washed with diethyl ether (2 × 50 mL), and evaporated to obtain the crude product. The obtained crude product was basicized with NaHCO3 (20 mL), extracted with DCM (3 × 50 mL), dried on 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. The fraction containing the desired product was recovered and lyophilized to obtain (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]hexane-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]hexane-2-yl)-1-((1S,3R,5S)-3-(2-hydroxyethoxy)adamantan-1-yl)-2-oxoethyl)carbamate (36 mg, 1.0 equivalent) in dichloromethane (0.21 mL), triethylamine (0.04 mL, 4.0 equivalents) and 4-nitrophenyl chloroformate (17.1 mg in 0.13 mL of dichloromethane) were added at 0°C. After stirring at room temperature for 4 hours, the mixture was added at 0°C 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 equivalents) and triethylamine (0.08 mL, 6.0 equivalents) in acetonitrile (0.2 mL + 0.05 mL * 2 washes). The reaction mixture was stirred for 12 hours, after which saturated aqueous NaHCO3 solution was added. The mixture was then extracted three times with dichloromethane. The combined organic layer was washed with brine and concentrated under vacuum to obtain the crude product, which was used in the next step without further purification. The crude product was treated with trifluoroacetic acid (0.1 mL) and dichloromethane (0.2 mL). After stirring for 4 hours, volatile substances were removed under reduced pressure. Purification of the crude composition by C18 reverse-phase preparative HPLC yielded 2-(((1R,3S,5S)-3-((S)-1-amino-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexane-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 (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]hexane-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), sodium azide (0.86 g, 13.33 mmol) was added at room temperature, and the reaction mixture was stirred at 80 °C for 2 hours. After completion, the reaction mixture was quenched with ice-cold solid water, filtered through a Buchner funnel, and dried to obtain tert-butyl (4-(azidomethyl)benzyl)carbamate (1.5 g) as an off-white solid. TLC system: siRNA:petroleum ether (0.3:0.7); R f :0.4.

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

[0305] Triphenylphosphine (1.9 g, 7.43 mmol) was added to a stirred solution of tert-butyl (4-(azidomethyl)benzyl)carbamate (1.3 g, 4.95 mmol) in methanol (15 mL), and the reaction mixture was stirred at 80°C for 2 hours. After completion, the reaction mixture was evaporated and purified by grace column chromatography using 8-10% MeOH in DCM to obtain tert-butyl (4-(aminomethyl)benzyl)carbamate (0.9 g) as a gum-like substance. 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)aminomethyl)benzyl)carbamate: [ka]

[0306] A mixture 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) was stirred in methanol (26 mL). Acetic acid (0.8 mL) was added at 0°C, and the resulting mixture was stirred at room temperature for 20 minutes. Then, sodium borocyanohydride (0.49 g, 7.81 mmol) was 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 fraction was evaporated to obtain tert-butyl(4-((bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)aminomethyl)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)azandiyl)bis(hexane-1,2,3,4,5-pentaol) 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), 4 M hydrochloric acid in 1,4-dioxane (8.0 mL) was added at 0°C, and the mixture was stirred at room temperature for 2 hours. After the completion of the reaction (monitored by LC-MS), the reaction mixture was evaporated, yielding (2R,2'R,3R,3'R,4R,4'R,5S,5'S)-6,6'-((4-(aminomethyl)benzyl)azandiyl)bis(hexane-1,2,3,4,5-pentaol) hydrochloride (0.95 g, crude) as a white gum-like solid.

[0309] Step 5 tert-butyl((1S)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexane-2-yl)-1-((1S,3R,5S)-3-(2-(((4-nitrophenoxy)carbonyl)oxy)xyethoxy)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]hexane-2-yl)-1-((1S,3R,5S)-3-(2-hydrochyethoxy)cyadamantan-1-yl)-2-oxoethyl)carbamate (0.8 g, 1.74 mmol) in dichloromethane (10 mL), triethylamine (0.9 mL, 1.91 mmol) was added, followed by the addition of 4-nitrophenylcarbonochloride (0.38 g, 6.96 mmol) in dichloromethane (2 mL) at -10°C. The reaction mixture was stirred at room temperature for 2 hours. After the reaction was complete, the reaction mixture was directly incorporated into the next step.

[0311] Process 6.2-(((1R,3S,5S)-3-((S)-1-((tert-butoxycarbonyl)amino)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexane-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] TEA (1.3 mL, 9.60 mmol) was added at 0°C to a stirred solution of (2R,2'R,3R,3'R,4R,4'R,5S,5'S)-6,6'-((4-(aminomethyl)benzyl)azandiyl)bis(hexane-1,2,3,4,5-pentaol) hydrochloride (0.96 g, 1.92 mmol) in ACN (10 mL). Next, tert-butyl((1S)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexane-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 the reaction was complete, the reaction mixture was concentrated and purified by RP C18 column chromatography using 25% MeOH and 0.1 M FA in water. The fraction containing the desired product was recovered and evaporated to obtain 2-(((1R,3S,5S)-3-((S)-1-((tert-butoxycarbonyl)amino)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexane-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 (0.8g) 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]hexane-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] 2-(((1R,3S,5S)-3-((S)-1-((tert-butoxycarbonyl)amino)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexane-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) and DCM (8 mL) were stirred together. TFA (4 mL) was added dropwise at 0°C, and the reaction mixture was stirred at room temperature for 1 hour. After the completion of the reaction (monitored by LC-MS), the reaction mixture was concentrated, washed with diethyl ether (2 x 100 mL), and evaporated to obtain the crude product. The crude product obtained was purified using RP SUNFIRE-C18 (150*19*5μ) with 20% ACN and 0.1% TFA in water. The fraction containing the desired product was recovered and lyophilized to obtain 2-(((1R,3S,5S)-3-((S)-1-amino)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexane-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]hexane-2-yl)-2-oxoethyl)adamantan-1-yl)oxy)ethyl(4-((ethyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)benzyl)carbamateformate (23): [ka]

[0315] Process 1: tert-butyl((1S)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexane-2-yl)-1-((1S,3R,5S)-3-(2-(((4-nitrophenoxy)carbonyl)oxy)xyethoxy)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]hexane-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), TEA (0.9 mL, 6.96 mmol) was added, followed by the dropwise addition of 4-nitrophenylcarbonochloride (0.38 g, 1.91 mmol) in DCM (0.5 mL) at -10°C, and the mixture was stirred at room temperature for 2 hours. After completion, the reaction mixture was incorporated into the next step to obtain tert-butyl((1S)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexane-2-yl)-1-((1S,3R,5S)-3-(2-(((4-nitrophenoxy)carbonyl)oxy)xyethoxy)adamantan-1-yl)-2-oxoethyl)carbamate (1.17g) as a yellow solution. TLC system: 100% ELISA, 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]hexane-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] TEA (1.5 mL, 11.22 mL) was added at 0°C to a stirred solution of tert-butyl(2R,3R,4R,5S)-6-((4-(aminomethyl)benzyl)(ethyl)amino)hexane-1,2,3,4,5-pentaol hydrochloride (0.81 g, 2.24 mmol) in ACN (10 mL). Then, tert-butyl((1S)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexane-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 hours. After the reaction was complete, the reaction mixture was concentrated and purified by RP C18 column chromatography using 27% ACN and 0.1 M FA in water. The fraction containing the desired product was recovered and evaporated to obtain 2-(((1R,3S,5S)-3-((S)-1-((tert-butoxycarbonyl)amino)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexane-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]hexane-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]hexane-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), TFA (10 mL) was added dropwise at 0°C, and the resulting mixture was stirred at room temperature for 1 hour. After completion (monitoring by LC-MS), the reaction mixture was concentrated and washed with diethyl ether (2 × 100 mL) to obtain the crude product. Next, the crude product was purified by RP chromatography using a C-18 column and 20% CAN:0.1M FA in water as the eluent. The fraction containing the desired product was recovered and lyophilized to obtain 2-(((1R,3S,5S)-3-((S)-1-amino)-2-((1S,3S,5S)-3-cyano-2-azabicyclo[3.1.0]hexane-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-aminopiperidine-1-yl)-7-(buta-2-in-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)phenyl)boronic acid (27): [ka]

[0320] Step 1(R)-(2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)phenyl)boronic acid: [ka]

[0321] To a stirred solution of tert-butyl(R)-(1-(7-(buta-2-in-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-8-yl)piperidine-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), K2CO3 (0.03 g, 0.24 mmol) was added, and the reaction mixture was stirred at room temperature for 16 hours. After the completion of the starting materials, the reaction mixture was diluted with ELISA (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 a crude mass. The crude residue was purified by reverse-phase preparative HPLC to obtain 0.06 g of (R)-(2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)phenyl)boronic acid as an off-white solid. [TLC system: Ԃ:petroleum ether (8:2); Rf value: 0.5].

[0322] Step 2(R)-(2-((8-(3-aminopiperidine-1-yl)-7-(buta-2-in-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)phenyl)boronic acid (27): [ka]

[0323] (R)-(2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)phenyl)boronic acid (0.06 g, 0.11 mmol) was stirred in 1,4-dioxane (0.6 mL). 4 M HCl in 1,4-dioxane (0.12 mL) was added dropwise at 0°C, and the reaction mixture was stirred at room temperature for 3 hours. After the starting material was complete, the reaction mixture was concentrated under reduced pressure and ground with diethyl ether to obtain 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] Hexyl(R)-2-((8-(3-aminopiperidine-1-yl)-7-(buta-2-in-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-6-fluoronicotinate (30) synthesis: [ka]

[0324] Step 1(R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-6-fluoronicotinic acid [ka]

[0325] To a stirred solution of methyl(R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-6-fluoronicotinate (0.9 g, 1.56 mmol) in THF (9 mL) and water (2 mL), LiOH.H2O (0.129 g, 3.08 mmol) was added at 0°C. The reaction mixture was stirred at room temperature for 4 hours. After the starting materials were removed, 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 obtain 0.72 g of compound (R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-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-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-6-fluoronicotinate: [ka]

[0327] A mixture of compound (R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-6-fluoronicotinic acid (0.72 g, 1.26 mmol) and C6H was added to a stirred solution in DMF (15 mL). 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 the completion of the starting materials, the reaction mixture was quenched with cold water, the precipitated solid was filtered off, and dried under vacuum to obtain 0.3 g of the compound hexyl(R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-6-fluoronicotinate as an off-white solid. [TLC system: siRNA: petroleum ether (1:1); R f Value: 0.8].

[0328] Step 3: Hexyl(R)-2-((8-(3-aminopiperidine-1-yl)-7-(buta-2-in-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-6-fluoronicotinate(30): [ka]

[0329] To a stirred solution of the compound hexyl(R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-6-fluoronicotinate (0.30 g, 0.46 mmol) in 1,4-dioxane (3 mL), 4 M 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 the starting material was fully digested, 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 0.181 g of the compound hexyl(R)-2-((8-(3-aminopiperidine-1-yl)-7-(buta-2-in-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-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] Hexyl(R)-2-((8-(3-aminopiperidine-1-yl)-7-(buta-2-in-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-6-(methylamino)nicotinic acid (31) synthesis: [ka]

[0330] Compound (R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-6-(methylamino)nicotinic acid (0.5 g, 0.86 mmol) was stirred in 3 mL of 1,4-dioxane. 4 M HCl in 2 mL of 1,4-dioxane was added at 0°C, and the reaction mixture was stirred at room temperature for 2 hours. After the starting material was fully digested, 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 0.157 g of the compound hexyl(R)-2-((8-(3-aminopiperidine-1-yl)-7-(buta-2-in-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-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] Hexyl(R)-2-((8-(3-aminopiperidine-1-yl)-7-(buta-2-in-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-6-(methylamino)nicotinate(32) synthesis: [ka]

[0331] Process 1(R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-6-(methylamino)nicotinic acid: [ka]

[0332] Compound (R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-6-(methylamino)nicotinic acid (1.0 g, 1.72 mmol, sharing the same intermediate as mMMT478) is stirred in DMF (10 mL) and C6H 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 the completion of the starting materials, the reaction mixture was quenched with cold water, the precipitated solid was filtered off, and dried under vacuum to obtain 0.7 g of compound (R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-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-aminopiperidine-1-yl)-7-(buta-2-in-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-6-(methylamino)nicotinate(32): [ka]

[0334] To a stirred solution of the compound hexyl(R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-6-(methylamino)nicotinate (0.41 g, 0.62 mmol) in 1,4-dioxane (4 mL), 4 M 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 the starting material was fully digested, 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 0.161 g of the compound hexyl(R)-2-((8-(3-aminopiperidine-1-yl)-7-(buta-2-in-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-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-aminopiperidine-1-yl)-7-(buta-2-in-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-6-((7-(4-((bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazole-1-yl)heptyl)amino)nicotinate (34): [ka]

[0335] To a solution of the 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-triazole-1-yl)heptyl)amino)-2-((7-(buta-2-in-1-yl)-8-((R)-3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)nicotinate (0.9 g, 0.70 mmol) in H2O (10 mL), concentrated HCl (2 mL) was added at 0°C, and the reaction mixture was stirred at room temperature for 2 hours. After the reaction was complete (LC-MS monitoring), the reaction mixture was concentrated under reduced pressure to obtain a crude mass. The crude product was purified by RP-HPLC to obtain the compound methyl 2-((8-((R)-3-aminopiperidine-1-yl)-7-(buta-2-in-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-6-((7-(4-((bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazole-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-aminopiperidine-1-yl)-7-(buta-2-in-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-6-((7-(4-((bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazole-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-triazole-1-yl)heptyl)amino)-2-((7-(buta-2-in-1-yl)-8-((R)-3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)nicotinic acid: [ka]

[0337] To a solution of the 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-triazole-1-yl)heptyl)amino)-2-((7-(buta-2-in-1-yl)-8-((R)-3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)nicotinate (0.350 g, 0.27 mmol) in MeOH (5 mL) and H2O (1 mL), NaOH (0.054 g, 1.35 mmol) was added, and the reaction mixture was stirred at room temperature for 16 hours. After the reaction was complete (LCMS monitoring), the reaction mixture was concentrated under reduced pressure to obtain a 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-triazole-1-yl)heptyl)amino)-2-((7-(buta-2-in-1-yl)-8-((R)-3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)nicotinic acid (0.3 g) as an off-white solid. The crude compound was then taken to the next step.

[0338] Step 2.2-((8-((R)-3-aminopiperidine-1-yl)-7-(buta-2-in-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-6-((7-(4-((bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazole-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-triazole-1-yl)heptyl)amino)-2-((7-(buta-2-in-1-yl)-8-((R)-3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)nicotinic acid (0.3 g, 0.24 mmol) in H2O (10 mL), concentrated HCl (0.6 mL) was added at 0°C, and the reaction mixture was stirred at room temperature for 2 hours. After the reaction was complete (LC-MS monitoring), the reaction mixture was concentrated under reduced pressure to obtain a crude mass. The crude product was purified by preparative HPLC to obtain compound 2-((8-((R)-3-aminopiperidine-1-yl)-7-(buta-2-in-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-6-((7-(4-((bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazole-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] Nonyl(R)-2-((8-(3-aminopiperidine-1-yl)-7-(buta-2-in-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-5-chlorobenzoate (37) synthesis: [ka]

[0340] Step 1: Isopropyl(R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-5-chlorobenzoate: [ka]

[0341] Compound (R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-5-chlorobenzoic acid (0.5 g, 0.85 mmol) was stirred in DMF (5 mL), to which 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 the starting material was complete, 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 the compound isopropyl(R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-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: siRNA:petroleum ether (3:7);R f Value: 0.7]

[0342] Step 2 synthesis of isopropyl(R)-2-((8-(3-aminopiperidine-1-yl)-7-(buta-2-in-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-5-chlorobenzoate (37): [ka]

[0343] To a stirred solution of compound isopropyl(R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-5-chlorobenzoate (0.4 g, 0.64 mmol) in 1,4-dioxane (4 mL), 4 M 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 the starting material was fully processed, the reaction mixture was concentrated under reduced pressure to obtain a crude mass. The crude compound was purified by reverse-phase preparative HPLC to obtain 0.19 g of the compound isopropyl(R)-2-((8-(3-aminopiperidine-1-yl)-7-(buta-2-in-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: 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-aminopiperidine-1-yl)-7-(buta-2-in-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-5-chlorobenzoate (38) was synthesized.

[0344] [Example 28] Synthesis of nonyl(R)-2-((8-(3-aminopiperidine-1-yl)-7-(buta-2-in-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-5-chlorobenzoate (39): [ka]

[0345] Step 1 Nonyl(R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-5-chlorobenzoate: [ka]

[0346] (R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-5-chlorobenzoic acid (0.35 g, 0.6 mmol) was stirred in DMF (3.5 mL), to which 1-bromonanone (0.15 g, 0.72 mmol) and K2CO3 (0.25 g, 1.8 mmol) were added, and the reaction mixture was stirred at room temperature for 16 hours. After the starting material was complete, 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-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-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: siRNA:petroleum ether (1:1); Rf value: 0.7].

[0347] Step 2 Nonyl(R)-2-((8-(3-aminopiperidine-1-yl)-7-(buta-2-in-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-5-chlorobenzoate (39) synthesis: [ka]

[0348] Nonyl(R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-5-chlorobenzoate (0.35 g, 0.49 mmol) was stirred in 1,4-dioxane (2 mL). 4 M HCl in 1,4-dioxane (2 mL) was added at 0°C, and the reaction mixture was stirred at room temperature for 3 hours. After the starting material was fully processed, the reaction mixture was concentrated under reduced pressure to obtain a crude mass. The crude compound was purified by reverse-phase preparative HPLC to obtain 0.13 g of nonyl(R)-2-((8-(3-aminopiperidine-1-yl)-7-(buta-2-in-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-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-aminopiperidine-1-yl)-7-(buta-2-in-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-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-triazole-1-yl)ethoxy)ethyl 2-((8-((R)-3-aminopiperidine-1-yl)-7-(buta-2-in-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-5-chlorobenzoate (41): [ka]

[0350] Step 1. 2-(2-bromoethoxy)ethyl(R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-5-chlorobenzoate: [ka]

[0351] (R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-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) were stirred in DMF (10 mL), to which K2CO3 (0.71 g, 5.13 mmol) was added, and the reaction mixture was stirred at room temperature for 16 hours. After the starting materials were completed, the reaction mixture was quenched with cold water, the precipitated solid was filtered off, and dried under vacuum to obtain 1.0 g of 2-(2-bromoethoxy)ethyl(R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-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: siRNA: petroleum ether (3:7); R f Value: 0.6].

[0352] Step 2. 2-(2-azidoethoxy)ethyl(R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-5-chlorobenzoate: [ka]

[0353] 2-(2-bromoethoxy)ethyl(R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-5-chlorobenzoate(3) (1.0 g, 1.36 mmol) was stirred in DMF (10 mL) and NaN3 (0.44 g, 6.79 mmol) was added. The reaction mixture was stirred at room temperature for 16 hours. After the starting material was complete, the reaction mixture was quenched with water and extracted with ethyl acetate (3 x 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 obtain the crude compound. Purification of the crude compound by column chromatography (using 100-200 mesh silica gel and 30% ethyl acetate in petroleum ether as an eluent) yielded 0.7 g of 2-(2-azidoethoxy)ethyl(R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-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: SiO: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-triazole-1-yl)ethoxy)ethyl 2-((7-(buta-2-in-1-yl)-8-((R)-3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-5-chlorobenzoate: [ka]

[0355] 2-(2-azidoethoxy)ethyl(R)-2-((7-(buta-2-in-1-yl)-8-(3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-5-chlorobenzoate (0. To a stirred solution of 3 g, 0.43 mmol) and (2R,3R,4R,5S)-6-(hexyl(propa-2-in-1-yl)amino)hexane-1,2,3,4,5-pentaol 5 (0.26 g, 0.86 mmol), CuSO4.5H2O (0.13 g, 0.52 mmol) and sodium ascorbate (0.13 g, 0.65 mmol) were added. The reaction mixture was stirred at room temperature for 8 hours. After the starting materials were fully digested, the reaction mixture was quenched with water and extracted with DCM (3 × 20 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 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-triazole-1-yl)ethoxy)ethyl 2-((7-(buta-2-in-1-yl)-8-((R)-3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-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-1,2,3triazole-1-yl)ethoxy)ethyl 2-((8-((R)-3-aminopiperidine-1-yl)-7-(buta-2-in-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-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-triazole-1-yl)ethoxy)ethyl 2-((7-(buta-2-in-1-yl)-8-((R)-3-((tert-butoxycarbonyl)amino)piperidine-1-yl)-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-1-yl)methyl)-5-chlorobenzoate (0.45 g, 0.45 mmol) in 1,4-dioxane (4.5 mL), 4 M 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 the starting material was removed, 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 41 (HCl salt) as an off-white solid. [TLC system: MeOH:DCM(1:9);R 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] 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) was added to a 10 mL solution of DCM with 4 M HCl in 1,4-dioxane at 0°C. The reaction mixture was then stirred at room temperature for 3 hours. After the reaction was complete, the reaction mixture was concentrated, the residue was basicized with saturated NaHCO3 solution, extracted with ethyl acetate (20 mL x 2), dried on anhydrous sodium sulfate, and evaporated to obtain the crude product. This was purified by silica gel (Davisil) column chromatography (using 0-8% MeOH in DCM as the eluent) to obtain 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-triazole-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-amino-4-(2,4,5trifluorophenyl)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-triazole-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] A stirred solution 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 (0.6 g, 0.847 mmol) and compound 6-(propa-2-in-1-ylamino)hexane-1,2,3,4,5-pentaol (0.371 g, 1.695 mmol) in t-BuOH (5 mL) and water (1 mL) is prepared by adding CuSO4. 4,5. H2O (0.254 g, 1.017 mmol) and sodium ascorbate (0.251 g, 1.271 mmol) were added. The resulting mixture was stirred at room temperature for 16 hours. After the reaction was complete, the reaction mixture was filtered. The filtrate was concentrated under reduced pressure to obtain the crude compound. The crude compound was purified by 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-triazole-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-((tert-butoxycarbonyl)amino)-4-(2,4,5trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate as a brown viscous 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 (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-triazole-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-amino-4-(2,4,5trifluorophenyl)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-triazole-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-((tert-butoxycarbonyl)amino)-4-(2,4,5trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (0.060 g, 0.064 mmol) in DCM (1 mL), 4 M HCl in 1,4-dioxane (0.6 mL) was added. The reaction mixture was stirred at room temperature for 3 hours. After the reaction was complete, the reaction mixture was filtered and concentrated to obtain the crude compound. The crude compound was purified by preparative HPLC to obtain 0.0157 g of 2-(2-(2-(4-((((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazole-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 (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-triazole-1-yl)ethoxy)ethoxy)ethyl7-((R)-3-amino-4-(2,4,5trifluorophenyl)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-pentaol: [ka]

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

[0365] Step 2.6-(ethyl(propa-2-in-1-yl)amino)hexane-1,2,3,4,5-pentaol: [ka]

[0366] To a solution of compound 6-(ethylamino)hexane-1,2,3,4,5-pentaol (2.0 g, 9.563 mmol), compound 3-bromopropa-1-yin (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 the reaction was complete, the reaction mixture was filtered, and the filtrate was concentrated to obtain 0.450 g of compound 6-(ethyl(propa-2-yin-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-triazole-1-yl)ethoxy)ethoxy)ethyl7-((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] A solution 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 (0.6 g, 0.847 mmol) and compound 6-(ethyl(propa-2-in-1-yl)amino)hexane-1,2,3,4,5-pentaol (0.420 g, 169 mmol) in t-BuOH (10 mL) and water (2 mL) is prepared by adding CuSO4. 4,5. H2O (0.254 g, 1.017 mmol) and sodium ascorbate (0.251 g, 1.271 mmol) were added. The resulting reaction mixture was stirred at room temperature for 16 hours. After the reaction was complete, 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-triazole-1-yl)ethoxy)ethoxy)ethyl7-((R)-3-((tert-butoxycarbonyl)amino)-4-(2,4,5trifluorophenyl)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 (Time / B %): 0 / 20, 1 / 20, 9 / 50, 14 / 50, 14.1 / 98, 18 / 98, 18.1 / 20, 21 / 20 Flow rate: 17 ml / min, Solubility: Acetonitrile + THF + Water Step 4.2 Synthesis of 2-(2-(2-(4-((ethyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazole-1-yl)ethoxy)ethoxy)ethyl7-((R)-3-amino-4-(2,4,5trifluorophenyl)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-triazole-1-yl)ethoxy)ethoxy)ethyl7-((R)-3-((tert-butoxycarbonyl)amino)-4-(2,4,5trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (0.240 g, 0.280 mmol) in DCM (3 mL), 4 M HCl in 1,4-dioxane (2.4 mL) was added. The reaction mixture was stirred at room temperature for 2 hours. After the reaction was complete, the reaction mixture was filtered and concentrated under reduced pressure to obtain the crude compound. The crude compound was purified by grinding with diethyl ether to obtain 175 mg of 2-(2-(2-(4-((ethyl((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazole-1-yl)ethoxy)ethoxy)ethyl7-((R)-3-amino-4-(2,4,5trifluorophenyl)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-triazole-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-triazole-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] Solution of 2-(2-(2-azidoethoxy)ethoxy)ethyl(R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate (460 mg, 0.65 mmol) in THF (3 mL). To this, 2R,3R,4R,5S)-6-(hexyl(propa-2-in-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) were added. The mixture was stirred at room temperature for 19 hours. The reaction mixture was filtered, and the filtrate was separated and purified by HPLC (Gemini-C18 150×21.2mm, 5um, mobile phase: ACN (0.1% FA)-H2O (0.1% FA), gradient: 20-70%) to obtain 2-(2-(2-(4-((hexyl(2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazole-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-triazole-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] 2-(2-(2-(4-((hexyl(2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazole-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 mixed with TFA (0.5 mL). The mixture was stirred at room temperature for 2 hours. Reaction mixture 40 o The mixture was concentrated under reduced pressure in 1C, and the residue was purified by preparative HPLC (Gemini-C18 150×21.2mm, 5um, mobile phase: ACN (0.1% TFA)-H2O (0.1% TFA), gradient: 20-70%) to obtain 2-(2-(2-(4-((hexyl(2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazole-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-triazole-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-(2,4,5trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate 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) was dissolved in ethyl acetate (1 L), to which HCl (4 M, 1 L) in ethyl acetate was added at room temperature. The mixture was stirred for 2 hours and concentrated to obtain the crude residue. This was redissolved in EA (1.00 L), quenched with an aqueous solution of NaHCO3 to adjust the pH to 8, the organic layer was separated, dried over Na2SO4, filtered, and the filtrate was concentrated to obtain 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-triazole-1-yl)ethoxy)ethoxy)ethyl7-((R)-3-amino-4-(2,4,5-trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate(46) Solutions of the 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 equivalent) and (2R,3R,4R,5S)-6-(hexyl(propa-2-in-1-yl)amino)hexane-1,2,3,4,5-pentaol (Int-1, 144 g, 474.63 mmol, 1.20 equivalent) were prepared in THF (2.00 L) and H2O (1.00 L). CuSO4.5H2O (99.0 g, 396 mmol, 1.00 equivalent) and sodium ascorbate (78.9 g, 398 mmol, 1.01 equivalent) were added to the mixture at 10-25°C, and the mixture was stirred at 25°C for 10 hours. Three batches of the mixture were diluted with NH3.H2O / NaCl aqueous solution = 1V / 2V (1.50 L), extracted with EA (1.50 L x 2), and washed with NH3.H2O / NaCl aqueous solution = 1V / 2V (1.50 L x 3) until the aqueous phase changed from colorless to blue. The organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The crude product was purified by reverse-phase HPLC (0.1% FA conditions), the mixture was concentrated to remove MeCN, then quenched with NaHCO3 to adjust the pH to 9, extracted with EA (10.0 L × 2), dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure to obtain 2-(2-(2-(4-((hexyl(2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazole-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, yield 25.9%, purity 96.3%) 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-triazole-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-triazole-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), tartaric acid (43.7 g, 291 mmol, 1.02 equivalents) in ethanol (1 L) was added. The mixture was stirred at 60°C for 5 hours. The mixture was concentrated under reduced pressure to obtain 2-(2-(2-(4-((hexyl(2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazole-1-yl)ethoxy)ethoxy)ethyl7-((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, yield 93.7%, purity 96.8%). 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-triazole-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(ethane-1-ol) (15.0 g, 99.88 mmol) in DCM (300 mL), Et3N (55.58 mL, 399.54 mmol) was added at 0°C, followed by tosyl chloride (76.16 g, 399.54 mmol). The reaction mixture was stirred at room temperature for 16 hours. After the reaction was complete, the reaction mixture was quenched with cold water (500 mL) and extracted with CH2Cl2 (2 × 500 mL). The combined organic layers were washed with cold water (500 mL) and brine (500 mL), dried on anhydrous Na2SO4, and concentrated under reduced pressure to obtain the crude compound. The crude compound was purified by column chromatography using silica gel (100-200 mesh) with 0-50% SiO in petroleum ether as the eluent to obtain 37.0 g of (ethane-1,2-diylbis(oxy))bis(ethane-2,1-diyl)bis(4-methylbenzenesulfonate) as an off-white solid. [TLC system: SiO:petroleum ether (4:6); Rf value: 0.6].

[0378] Step 21: Synthesis of 2-bis(2-azidoethoxy)ethane [ka]

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

[0380] Step 3.2-(2-(2-azidoethoxy)ethoxy)ethane-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), a solution of 1N HCl (17.6 mL) was added, followed by PPh3 (3.9 g, 14.98 mmol) in Et2O (22 mL). The reaction mixture was stirred at room temperature for 12 hours. After the reaction was complete, 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 obtain 2.0 g of 2-(2-(2-azidoethoxy)ethoxy)ethane-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]pyrazine-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-yl)carbamate [ka]

[0383] (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5trifluorophenyl)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)ethane-1-amine hydrochloride (1.66 g, 7.90 mmol) were stirred in DMF (30 mL), to which HATU (3.0 g, 7.90 mmol), followed by DIPEA (3.39 mL, 26.35 mmol), was added at 0°C. The reaction mixture was stirred at room temperature for 45 minutes. The progress of the reaction was monitored by TLC. After the completion of the reaction, the reaction mixture was quenched with cold water (2 x 50 mL) and extracted with ELISA (2 x 100 mL). The combined organic layers were washed with cold water (100 mL) and brine (100 mL), dried on anhydrous Na2SO4, and concentrated to obtain the crude compound. The crude compound was purified by silica gel column chromatography (Davisil) (using 0-70% ethyl phosphate in petroleum ether as the eluent) to obtain 3.0 g of tert-butyl(R)-(4-(1-((2-(2-(2-azidoethoxy)ethoxy)ethyl)carbamoyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazine-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-yl)carbamate as an off-white solid. [TLC system: ethyl phosphate: 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]pyrazine-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-yl)carbamate (2.4 g, 3.339 mmol) in CH2Cl2 (24 mL), 4 M HCl in 1,4-dioxane (24.0 mL) was added at 0°C. The reaction mixture was stirred at room temperature for 6 hours. After the reaction was complete, the reaction mixture was concentrated, the residue was basicized with saturated NaHCO3 solution, and extracted with 10% MeOH (2 x 200 mL) in DCM. The combined organic layers were dried over anhydrous Na2SO4 and concentrated to obtain 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 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-triazole-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-(2,4,5trifluorophenyl)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), CuSO4, 5.H2O (1.08 g, 4.352 mmol) and sodium ascorbate (1.07 g, 5.440 mmol) were added. The reaction mixture was stirred at room temperature for 2 hours. After the reaction was complete, the reaction mixture was filtered and concentrated to obtain the crude compound. Purification of the crude compound by RP preparative HPLC yielded 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-triazole-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 (19 x 250 mm) 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-(2,4,5trifluorophenyl)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-(2,4,5trifluorophenyl)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)(propa-2-in-1-yl)amino)hexane-1,2,3,4,5-pentaol (0.126 g, 0.329 mmol), CuSO4, 5H2O (0.05 g, 0.197 mmol) and sodium ascorbate (0.048 g, 0.246 mmol) were added. The reaction mixture was then stirred at room temperature for 16 hours. After the reaction was complete, the reaction mixture was filtered and concentrated to obtain the crude compound. The crude compound was purified by preparative HPLC to obtain 20 mg of 2-(2-(2-(4-((bis((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)amino)methyl)-1H-1,2,3-triazole-1-yl)ethoxy)ethoxy)ethyl 7-((R)-3-amino-4-(2,4,5trifluorophenyl)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), 5 μm. Mobile phase A: 10 mm ABC (pH) water. Mobile phase B: Acetonitrile.

[0390] Gradient (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]pyrazine-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butane-2-aminium(2R,3S)-3-carboxy-2,3-dihydroxypropanoate (tartrate)(49): [ka]

[0393] Step 1.2-Morpholinoethyl(R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylate: [ka]

[0394] (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) was stirred in 1,2-dichloroethane (10 mL) and triethylamine (0.37 g, 3.64 mmol). Bis(2-oxo-3-oxazolidinyl)phosphine chloride (0.46 g, 1.82 mmol) was added at 0°C, and the resulting mixture was stirred at 0°C for 1 hour. Then, 2-morpholinoethane-1-ol (0.24 g, 1.82 mmol) was added at 0°C, and the reaction mixture was stirred at room temperature for 16 hours. After completion, the reaction mixture was diluted with water (10 mL) and extracted with dichloromethane (2 × 50 mL). The combined organic layer was dried on anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude product. The obtained crude residue was then purified by column chromatography using silica gel (230-400 mesh) with a gradient of 4% methanol in dichloromethane to obtain 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), a 4.0 M hydrogen chloride solution in dioxane (5.1 mL) was added 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 then purified by RP preparative HPLC under the following conditions. Column / Dimensions: X-BRIDGE-C18 (19*250) 5um, Mobile Phase A: 10mM ammonium bicarbonate in water, Mobile Phase B: Acetonitrile, Gradient (Time / B %): 0 / 5, 1 / 5, 8 / 40, 13.5 / 40, 13.51 / 100, 18 / 100, 18.1 / 5, 20.5 / 5, Flow Rate: 18 ml / min. The desired fraction was lyophilized to obtain 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, 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]pyrazine-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butane-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 desalinated water (2.5 mL) was sonicated to obtain a clear solution. The resulting solution was freeze-dried to obtain (R)-4-(1-((2-morpholinoethoxy)carbonyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazine-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]pyrazine-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butane-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]pyrazine-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-(2,4,5trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylic acid (0.4 g, 0.73 mmol), 2-(2-morpholinoethoxy)ethane-1-amine (0.127 g), and (1-[bis(dimethylamino)methylene]-1H-1,2,3triazolo[4,5-b]pyridinium 3-oxidehexafluorophosphate (0.334 g, 0.88 mmol)) in N,N-dimethylformamide (4 mL), N,N-diisopropylethylamine (0.47 g, 3.65 mmol) was slowly added at 0°C, and the reaction mixture was stirred at room temperature for 4 hours. After completion, the reaction mixture was mixed. The mixture was diluted with water (30 mL) and extracted with ethyl acetate (3 × 100 mL). The combined organic layer was dried on anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude product. The crude residue was then purified by column chromatography using silica gel (230-400 mesh) with a gradient of 4% methanol in dichloromethane to obtain tert-butyl(R)-(4-(1-((2-(2-morpholinoethoxy)ethyl)carbamoyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazine-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]pyrazine-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), 4.0 M HCl in dioxane (4 mL) was added 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 obtain the crude product. The obtained crude product was then purified by RP preparative HPLC under 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 / B %): 0 / 10, 1 / 10, 10 / 40, 15.5 / 1 / 6 / 1.5, 15.6 / 100, 18 / 100, 18.1 / 10, 20 / 10, Flow Rate: 17 ml / min. The desired fraction was lyophilized to obtain (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, 46.6%) as a colorless gum. TLC system. MeOH:DCM(1:9);R f :0.05. Step 3(R)-4-(1-((2-(2-morpholinoethoxy)ethyl)carbamoyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazine-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butane-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), L(+)-tartaric acid (0.039 g, 0.26 mmol) was added, and the mixture was stirred at room temperature for 6 hours. The solvent was evaporated under reduced pressure, and the residue was then washed with n-pentane (2 × 5 mL) and redissolved in desalted water (1.4 mL). The resulting solution was freeze-dried to obtain (R)-4-(1-((2-(2-morpholinoethoxy)ethyl)carbamoyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazine-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]pyrazine-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butane-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] (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) was stirred in 1,2-dichloroethane (6 mL) and triethylamine (0.165 g, 1.63 mmol). Bis(2-oxo-3-oxazolidinyl)phosphine chloride (0.207 g, 0.817 mmol) was added at 0°C, and the resulting mixture was stirred at 0°C for 1 hour. 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 hours. After completion, the reaction mixture was diluted with water (10 mL) and extracted with dichloromethane (2 × 30 mL). The combined organic layer was dried on anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude product. The obtained crude residue was then purified by column chromatography using silica gel (230-400 mesh) with a gradient of 4% methanol in dichloromethane to obtain 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), 4.0 M of dioxane (2 mL) was added 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 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 / B%): 0.01 / 25, 1 / 25, 9 / 40, 13 / 40, 13.1 / 100, 18 / 100, 18.1 / 25, 20 / 25, Flow rate: 18 ml / min. The desired fraction was lyophilized to obtain 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]pyrazine-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butane-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), L(+)-tartaric acid (0.032 g, 0.216 mmol) was added, and the mixture was stirred at room temperature for 6 hours. The reaction mixture was concentrated and washed with n-pentane (2 × 5 mL). The resulting residue was dissolved in water and freeze-dried to obtain (R)-4-(1-((3-morpholinopropoxy)carbonyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazine-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]pyrazine-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butane-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-(2,4,5trifluorophenyl)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, to which bis(2-oxo-3-oxazolidinyl)phosphine chloride (0.28 g, 1.09 mmol) was added at 0°C, and the resulting mixture was stirred at 0°C for 1 hour. Then, 2-(2-morpholinoethoxy)ethane-1-ol (0.191 g, 1.09 mmol) was added at 0°C, and the reaction mixture was stirred at room temperature for 16 hours. After completion, the reaction mixture was diluted with water (10 mL) and extracted with dichloromethane (2 × 30 mL). The combined organic layer was dried on anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude product. The obtained crude residue was then purified by column chromatography using silica gel (230-400 mesh) with a gradient of 4% methanol in dichloromethane to obtain 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] 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) was stirred in 1,4-dioxane (3.2 mL). 4.0 M HCl in dioxane (3.2 mL) was added 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 obtain the crude product. The crude product was then purified by reverse-phase preparative HPLC under 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 / B %): 0 / 5, 1 / 5, 12 / 75, 14 / 75, 14.05 / 98, 16.50 / 98, 16.51 / 5, 19 / 5, Flow Rate: 18 ml / min. The desired fraction was lyophilized to obtain 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, 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]pyrazine-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butane-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 desalinated water (1.4 mL) was sonicated to obtain a clear solution. The resulting solution was freeze-dried to obtain (R)-4-(1-((2-(2-morpholinoethoxy)ethoxy)carbonyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazine-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]pyrazine-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butane-2-aminium(2R,3R)-3-carboxy-2,3-dihydroxypropanoate (tartrate)(53): [ka]

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

[0410] (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) was stirred in 1,2-dichloroethane (20 mL) and triethylamine (1.27 g, 9.1 mmol). Bis(2-oxo-3-oxazolidinyl)phosphine chloride (0.93 g, 3.64 mmol) was added at 0°C, and the resulting mixture was stirred at 0°C for 1 hour. 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 hours. After completion, the reaction mixture was diluted with water (10 mL) and extracted with dichloromethane (2 × 30 mL). The combined organic layer was dried on anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude product. The obtained crude residue was then purified by column chromatography using silica gel (230-400 mesh) with a gradient of 4% methanol in dichloromethane to obtain 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-Molfolinopentyl(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), 4.0 M HCl in dioxane (6 mL) was added 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 crude product was then purified by RP preparative HPLC under 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 / B%): 0 / 20, 1 / 20, 12 / 50, 16.5 / 50, 16.6 / 100, 19 / 100, 19.1 / 20, 21 / 20, Flow rate: 18 ml / min. The desired fraction was freeze-dried to obtain 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 (0.29 g, 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]pyrazine-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butane-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 desalinated water (1.9 mL) was sonicated for 2 minutes to obtain a clear solution. The resulting solution was freeze-dried to obtain (R)-4-(1-(((5-morpholinopentyl)oxy)carbonyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazine-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butane-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-morpholinbutoxy)carbonyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazine-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butane-2-aminium(2R,3R)-3-carboxy-2,3-dihydroxypropanoate (tartrate)(54): [ka]

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

[0414] (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) was stirred in 1,2-dichloroethane (10 mL) and triethylamine (0.37 g, 3.64 mmol). Bis(2-oxo-3-oxazolidinyl)phosphine chloride (0.35 g, 1.36 mmol) was added at 0°C, and the mixture was stirred at 0°C for 1 hour. 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 hours. After completion, the reaction mixture was diluted with water (10 mL) and extracted with dichloromethane (2 × 30 mL). The combined organic layer was dried on anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude product. The crude residue was then purified by column chromatography using silica gel (230-400 mesh) with 4% methanol in dichloromethane as the eluent, yielding 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-Molfolinobutyl(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), 4.0 M HCl in dioxane (4.5 mL) was added 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 crude product was then purified by RP preparative HPLC under 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 / B%): 0 / 20, 2 / 20, 10 / 45, 14 / 45, 14.10 / 100, 16 / 100, 16.10 / 20, 18 / 20, Flow Rate: 17 ml / min. The desired fraction was lyophilized to obtain 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 (0.16 g, 41.7%) as a colorless gum. TLC system. MeOH:DCM(1:9);R f :0.05. Step 3(R)-4-(1-((4-morpholinbutoxy)carbonyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazine-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butane-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 desalinated water (1 mL) was sonicated to obtain a clear solution. The resulting solution was freeze-dried to obtain (R)-4-(1-((4-morpholinobtoxic)carbonyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazine-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butane-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]pyrazine-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butane-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]pyrazine-7(8H)-yl)-4-oxo-1-(2,4,5-trifluorophenyl)butan-2-yl)carbamate: [ka]

[0418] (R)-7-(3-((tert-butoxycarbonyl)amino)-4-(2,4,5trifluorophenyl)butanoyl)-3-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine-1-carboxylic acid (0.5g, 0.91 mmol) and 2-methyl-2-azaspiro[3.3]heptane-6-amine salt in N,N-dimethylformamide (5mL) To a stirred solution of (0.18 g, 0.91 mmol) of the salt (1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (0.415 g, 0.1.09 mmol)), N,N-diisopropylethylamine (0.59 g, 4.55 mmol) was slowly added at 0°C, and the reaction mixture was stirred at room temperature for 4 hours. After completion, the reaction The mixture was diluted with water (30 mL) and extracted with ethyl acetate (3 × 100 mL). The combined organic layer was dried on anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude product. The crude residue was then purified by column chromatography on silica gel (230-400 mesh) using 4% methanol in dichloromethane as the eluent to obtain tert-butyl(R)-(4-(1-((2-methyl-2-azaspiro[3,3]heptan-6-yl)carbamoyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazine-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-(2,4,5trifluorophenyl)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]pyrazine-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), 4.0 M HCl in dioxane (3 mL) was added 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 obtain the crude product. The obtained crude product was then purified by RP preparative HPLC under 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 / B%): 0 / 10, 1 / 10, 10 / 40, 16 / 63.2, 16.1 / 100, 19 / 100, 19.1 / 10, 22 / 10, Flow Rate: 17 ml / min. The desired fraction was freeze-dried to obtain (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 (0.09 g, 35.4%) as an off-white solid. TLC system. MeOH:DCM(1:9);R 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]pyrazine-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) was added to dichloromethane (4 mL), and the reaction mixture was stirred at room temperature for 6 hours. The reaction mixture was concentrated, the residue was washed with n-pentane (2 x 5 mL), and the residue was dissolved in desalted water (3 mL). The resulting solution was freeze-dried to obtain (R)-4-(1-((2-methyl-2-azaspiro[3,3]heptan-6-yl)carbamoyl)-3-(trifluoromethyl)-5,6-dihydroimidazo[1,5-a]pyrazine-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 were obtained using the DPP4 Activity Assay Kit (Sigma-Aldrich, MAK088) according to the manufacturer's instructions. Briefly, 10 μL of DPP4 assay buffer per well was added to a low-volume 384-well plate before transferring 10 μL of the test compound dissolved in DPP4 assay buffer. 5 μL of master reaction mix containing a fluorescent substrate that becomes fluorescent upon enzymatic cleavage was added to each well. Fluorescence intensity measurements were recorded at 1-minute intervals over a 20-minute process using an Envision multimode plate reader (PerkinElmer). The results are shown in Table 4 below.

[0421] AEC2 proliferation assay. Primary human AEC2 cells were plated at a density of 1,500 cells per well in 50 μL of small airway epithelial cell proliferation medium (Lonza) containing 5% BPE and free of EGF and retinoic acid, in a black 384-well plate (Greiner) coated with 10 ug / mL laminin (Life Technologies). Then, 100 nL of the test compound dissolved in DMSO was delivered using a Biomek FX instrument (Beckman Coulter) fitted with a pin tool head (V&P Scientific). After proliferation at 37°C for 96 hours, the cells were fixed with 4% paraformaldehyde, washed three times with PBS, and then immunostained for KI-67 positivity overnight at 4°C (1:1000, Abcam, ab15580). After three further washes, the cells were incubated with a secondary AlexaFluor conjugate secondary antibody at room temperature for 1 hour, and then exposed to 10 μg / mL Hoechst 33342 (Life Technologies). The plates were sealed, and quantitative high-content imaging was performed using a CellInsight CX5 HCS instrument (ThermoFisher). The ACE2 growth concentration curve for compound 46 is shown in Figure 1. [Table 6] TIFF0007927730000233.tif201153TIFF0007927730000234.tif190154TIFF0007927730000235.tif22 7154TIFF0007927730000236.tif133154TIFF0007927730000237.tif184154TIFF0007927730000238.t if161153TIFF0007927730000239.tif235154TIFF0007927730000240.tif149153TIFF00079277300002 41.tif199153TIFF0007927730000242.tif202153TIFF0007927730000243.tif215154TIFF00079277300 00244.tif198153TIFF0007927730000245.tif167153TIFF0007927730000246.tif209153TIFF0007927 730000247.tif163154TIFF0007927730000248.tif167153TIFF0007927730000249.tif190154TIFF000 7927730000250.tif202154TIFF0007927730000251.tif200154TIFF0007927730000252.tif200154TIF F0007927730000253.tif188154TIFF0007927730000254.tif218154TIFF0007927730000255.tif158154

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

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

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

[0425] [Example 47] In vivo efficacy study Experimental methods for the ALI model Acute lung injury was induced in mice using LPS derived from E. coli I111:B4(Sigma). Female C57BL / 6J mice aged 9–11 weeks with matched body weight (19 g–22 g) were selected to be used as an ALI model.

[0426] For oral delivery, the DPP4 inhibitor was dissolved in PBS to obtain a clear solution. The vehicle control or DPP4 inhibitor was administered at 10 ml / kg via oral force-feeding once or twice daily, selected based on the PK profile. For intratracheal delivery of the DPP4 inhibitor, the compound was dissolved in PBS to obtain a clear solution. The vehicle control or DPP4 inhibitor was administered at 2 ml / kg every other day via a 22 g flexible catheter.

[0427] Mice were intratracheally injected into the lungs on day 0 with either 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 placebo group. Mice were administered either a DPP4 inhibitor or a vehicle control, starting one day before (-1 day) the LPS injection.

[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 readability, the total protein content in BALF was quantified using a BCA assay. Lung inflammation and damage were assessed using H&E staining.

[0430] Experimental methods for the bleomycin model Pulmonary fibrosis was induced in mice using bleomycin (Hospira). Male C57BL / 6J mice, 10–12 weeks old, with matched body weight (24–28 grams), were selected for use in the bleomycin model.

[0431] For oral delivery, the DPP4 inhibitor was dissolved in PBS to obtain a clear solution. The vehicle control or DPP4 inhibitor was administered at 10 ml / kg via oral force-feeding once or twice daily, selected based on the PK profile. For intratracheal delivery of the DPP4 inhibitor, the compound was dissolved in PBS to obtain a clear solution. The vehicle control or DPP4 inhibitor was administered at 2 ml / kg every four days via a 22 g flexible catheter.

[0432] 0.5 U / kg of bleomycin or PBS (in the placebo group) was intratracheally injected into the lungs of mice on day 0. A DPP4 inhibitor or vehicle control was administered to the mice starting one day before the 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] Body weight was measured daily for readability. Total protein content in BALF was quantified using a BCA assay. Pulmonary fibrosis was assessed using Masson's trichrome staining.

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

[0436] In a rodent bleomycin model, we decided to further evaluate the efficacy of compound 46 in combination with the standard therapeutic agent nintedanib (Figure 4). The combination of compound 46 and nintedanib showed impressive synergistic efficacy, as determined by Bliss independence calculations on BALF protein content and Ashcroft score metrics for fibrosis severity.

[0437] To understand which cell types proliferate in response to compound 46 treatment, single-cell RNA sequencing was performed from mouse lung cells. After treatment with the compound (0.5 mg / kg IT, animals sacrificed 2 and 4 days after administration), only circulating and transitional AEC2 cells were induced to proliferate among other cell populations, including other epithelial cell types, but excluding actively proliferating immune cells (Figure 5A, Figure 5B). The data further demonstrate time-dependent accumulation of AEC2 cells after treatment with the compound. In one embodiment, the present invention provides the following. [Item 1] A compound of formula (I): TIFF0007927730000257.tif45142 During the ceremony Each ---, if optionally present, represents a single bond that forms a condensed cyclopropyl ring; L 1A However, -NHCH 2 - or -CH(NH 2 )-and; X 1 is -O-, -S-, -S(O)-, S(O) 2 - and -NH- are selected; L 1B However, one or more -CH 2 - may be independently substituted by a part selected from -O-, -C(O)-, and -NH-. 2 -C 12 -It is alkyl; Z 1 However, H, C 6 -C 10 -aryls, and 5- to 10-membered heteroaryls (1 to 4 heteroaryl members are independently selected from N, O, and S); m1 is Z 1 If H, then it is 0, and Z 1 It is an integer that is 1 if it is not H; n1 is an integer selected from 0, 1, 2, and 3; R 1 However, H, C 1 -C 10 -alkyl and -C 1 -C 10 -alkyl-(C 6 -C 10 Selected from -aryl groups, which may be substituted with 1 to 6 -OH groups; R 2 However, C substituted with 1 to 6 -OH groups 1 -C 10 -alkyl compounds; or A compound of formula (II): TIFF0007927730000258.tif35155 During the ceremony W is either CH or N; o is an integer selected from 1, 2, and 3; R 3 However, C 1 -C 6 -alkyl, C 1 -C 6 -hydroxyalkyl and -(CH 2 CH 2 O) x H(x is an integer selected from 1, 2, 3, 4, and 5); R 4 However, C2 -C 8 -is Alkinnil; R 5a 、R 5b 、R 5c , and R 5d However, H, C 1 -C 6 -alkyl, halo, -NR A R B (R A and R B H and C 1 -C 10 -Selected independently of 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, and one or more -CH 2 - may be independently substituted by a moiety selected from -O-, -C(O)-, and -NH-, -C(O)-L 2 -Z 2 -[(CH 2 ) n2 -NR 6 R 7 ] m2 Selected independently of; Z 2 However, H, C 6 -C 10 - Selected from aryls and 5- to 10-membered heteroaryls (1- to 4 heteroaryl members are independently selected from N, O, and S); R 6 However, H, C 1 -C 10 -alkyl and -C 1 -C 10 -alkyl-(C 6 -C 10 Selected from -aryl groups, which may be substituted with 1 to 6 -OH groups; R 7 However, C substituted with 1 to 6 -OH groups 1 -C10 -It is alkyl; m2 is Z 1 If H, then it is 0, and Z 1 It is an integer that is 1 if it is not H; n2 is an integer selected from 0, 1, 2, and 3; R 5a 、R 5b 、R 5c , and R 5d At least one of them is not H; If W is CH, then R 5a and R 5d However, compounds not selected from -C(O)OH, -C(O)OMe, and -C(O)OEt; or A compound of formula (III): TIFF0007927730000259.tif40154 During the ceremony X 3 However, it is -O- or -NH-; L 3 However, bonding, or one or more -CH groups 2 - may be independently substituted by a part selected from -O-, -C(O)-, and -NH-. 2 -C 12 -It is alkyl; Z 3 However, H, -N 3 ,、C 6 -C 10 - Selected from aryl, 5-10 membered heteroaryl (1-4 heteroaryl members independently selected from N, O, and S), and (3-14 membered heterocycloalkyl (1-4 ring members independently selected from N, O, and S), Heteroaryl and heterocycloalkyl compounds are halos, NO 2 OH, CN, and C 1 -C 6 - May be substituted with 1 to 6 substituents selected from the group consisting of haloalkyl groups; m3, Z 3 is H or -N 3 In this case, it is 0, and Z 3 is H or -N 3 An integer that is 1 in all other cases; n3 is an integer selected from 0, 1, 2, and 3; R 8 However, H, C 1 -C 10 -alkyl and -C1 -C 10 -alkyl-(C 6 -C 10 Selected from -aryl groups, which may be substituted with 1 to 6 -OH groups; R 9 However, C substituted with 1 to 6 -OH groups 1 -C 10 -It is alkyl; R 10 However, C 1 -C 6 -It is a haloalkyl; Each R 11 However, H, C 1 -C 6 -Selected independently from 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; Compounds where q3 is an integer selected from 0, 1, 2, and 3; or a pharmaceutically acceptable salt thereof However, the following compounds are excluded. TIFF0007927730000260.tif74158TIFF0007927730000261.tif241132TIFF0007927730000262.tif113135 [Item 2] A compound of formula (I) as described in item 1, or a pharmaceutically acceptable compound thereof. [Item 3] L 1A ga-NHCH 2 The compounds listed in item 2 or any pharmaceutically acceptable compounds thereof. [Item 4] L 1A -CH(NH 2 )- The compound listed in item 2 or any pharmaceutically acceptable compound thereof. [Item 5] X 1 A compound or a pharmaceutically acceptable compound described in any one of items 2 to 4, wherein the compound is O. [Item 6] Z 1 A compound or a pharmaceutically acceptable compound described in any one of items 2 to 5, wherein the compound is H. [Item 7] Z1 C 6 -C 10 - A compound described in any one of items 2 to 5, which is an aryl or a 5- to 10-membered heteroaryl (1 to 4 heteroaryl members are independently selected from N, O, and S), or any pharmaceutically acceptable compound thereof. [Item 8] Z 1 A compound or a pharmaceutically acceptable compound described in any one of items 2-5 and 7, wherein the compound is phenyl. [Item 9] Z 1 A compound or any pharmaceutically acceptable compound described in any one of items 2-5 and 7, wherein triazolyl is used. [Item 10] A compound or any pharmaceutically acceptable compound described in any one of items 7-9, wherein n1 is 1 or 2. [Item 11] R 1 However, C may be substituted with 1 to 6 -OH groups. 1 -C 10 -A compound that is alkyl, as described in any one of items 7-10, or any pharmaceutically acceptable compound thereof. [Item 12] R 1 C 1 -C 6 -A compound that is alkyl, as described in any one of items 7-11, or any pharmaceutically acceptable compound thereof. [Item 13] R 2 However, C is substituted with 1 to 5 -OH groups. 2 -C 6 -A compound that is alkyl, as described in any one of items 7-12, or any pharmaceutically acceptable compound thereof. [Item 14] R 2 However, C is substituted with 3 to 5 -OH groups. 2 -C 6 -A compound that is alkyl, as described in any one of items 7-13, or any pharmaceutically acceptable compound thereof. [Item 15] R 2 but: TIFF0007927730000263.tif26152 The compound described in any one of items 7-14, or any pharmaceutically acceptable compound thereof. [Item 16] Z 1 However, C 6 -C 10 -Aryl or 5-10 member heteroaryl (1-4 heteroaryl members are independently selected from N, O, and S); n1 is either 1 or 2; R 1 However, C 1 -C 10 -It is alkyl; R 2 However, C substituted with 1 to 5 -OH groups 2 -C 6 - Compounds that are alkyl, as listed in item 2. [Item 17] A compound listed in item 1 or 2, or a pharmaceutically acceptable compound, selected from the table below. TIFF0007927730000264.tif69159TIFF0007927730000265.tif225162TIFF0007927730000266.tif236162 [Item 18] A compound of formula (II) as described in item 1, or a pharmaceutically acceptable compound thereof. [Item 19] A compound listed in item 18, or any pharmaceutically acceptable compound, wherein W is CH. [Item 20] A compound listed in item 18, or any pharmaceutically acceptable compound, wherein W is N. [Item 21] R 5b and R 5d A compound or a pharmaceutically acceptable compound described in any one of items 18-20, wherein the compound is H. [Item 22] R 5a -C(O)OH or -C(O)OR A The compound described in any one of items 18-21 or any pharmaceutically acceptable compound thereof. [Item 23] R 5a -C(O)-L 2 -Z 2 -[(CH 2 ) n2 -NR 6 R 7 ] m2 The compound described in any one of items 18-21 or any pharmaceutically acceptable compound thereof. [Item 24] Z 2 C 6 -C10 - A compound described in any one of items 18-23, or any pharmaceutically acceptable compound thereof, which is an aryl or a 5- to 10-membered heteroaryl (1 to 4 heteroaryl members are independently selected from N, O, and S). [Item 25] Z 2 A compound described in any one of items 18-24, or any pharmaceutically acceptable compound thereof, wherein the compound is phenyl or triazolyl. [Item 26] Z 2 A compound or any pharmaceutically acceptable compound described in any one of items 18-25, wherein the compound is triazolyl. [Item 27] A compound or any pharmaceutically acceptable compound described in any one of items 18-26, wherein n2 is 1 or 2. [Item 28] R 6 However, C may be substituted with 1 to 6 -OH groups. 1 -C 10 -A compound that is alkyl, as described in any one of items 18-27, or any pharmaceutically acceptable compound thereof. [Item 29] R 6 C 1 -C 6 -A compound that is alkyl, as described in any one of items 18-28, or any pharmaceutically acceptable compound thereof. [Item 30] R 7 However, C is substituted with 1 to 5 -OH groups. 2 -C 6 -A compound that is alkyl, as described in any one of items 18-29, or any pharmaceutically acceptable compound thereof. [Item 31] R 7 However, C is substituted with 3 to 5 -OH groups. 2 -C 6 -A compound that is alkyl, as described in any one of items 18-30, or any pharmaceutically acceptable compound thereof. [Item 32] R 7 but: TIFF0007927730000267.tif26155 The compound described in any one of items 18-31 or any pharmaceutically acceptable compound thereof. [Item 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 However, C 6 -C 10 -The aryl or 5-10 member heteroaryl (1-4 heteroaryl members are independently selected from N, O, and S); n2 is either 1 or 2; R 6 However, C 1 -C 10 -It is alkyl; R 7 However, C substituted with 1 to 5 -OH groups 2 -C 6 - Compounds that are alkyl, as listed in item 18. [Item 34] A compound listed in item 1 or 18, or any pharmaceutically acceptable compound, selected from the table below. TIFF0007927730000268.tif210151 [Item 35] A compound of formula (III) as described in item 1, or a pharmaceutically acceptable compound thereof. [Item 36] X 3 A compound listed in item 35 or any pharmaceutically acceptable compound thereof, wherein the compound is O. [Item 37] X 3 A compound listed in item 35 or any pharmaceutically acceptable compound thereof, wherein the compound is -NH-. [Item 38] Z 3 ga-N 3 The compound described in any one of items 35-37, or any pharmaceutically acceptable compound thereof. [Item 39] Z 3 C 6 -C 10 - A compound described in any one of items 35-37, or any pharmaceutically acceptable compound thereof, which is an aryl or a 5- to 10-membered heteroaryl (1 to 4 heteroaryl members are independently selected from N, O, and S). [Item 40] A compound described in any one of items 35-39, or any pharmaceutically acceptable compound thereof, wherein p3 and q3 are each 1. [Item 41] Z 3 A compound described in any one of items 35-40, or any pharmaceutically acceptable compound thereof, wherein the compound is phenyl or triazolyl. [Item 42] Z 3 A compound described in any one of items 35-41, or any pharmaceutically acceptable compound thereof, wherein triazolyl is used. [Item 43] A compound or any pharmaceutically acceptable compound described in any one of items 35-42, wherein n3 is 1 or 2. [Item 44] R 8 However, C may be substituted with 1 to 6 -OH groups. 1 -C 10 -A compound that is alkyl, as described in any one of items 35-43, or any pharmaceutically acceptable compound thereof. [Item 45] R 8 C 1 -C 6 -A compound that is alkyl, as described in any one of items 35-44, or any pharmaceutically acceptable compound thereof. [Item 46] R 9 However, C is substituted with 1 to 5 -OH groups. 2 -C 6 -A compound that is alkyl, as described in any one of items 35-45, or any pharmaceutically acceptable compound thereof. [Item 47] R 9 However, C is substituted with 3 to 5 -OH groups. 2 -C 6 -A compound that is alkyl, as described in any one of items 35-46, or any pharmaceutically acceptable compound thereof. [Item 48] R 9 but: TIFF0007927730000269.tif27153 The compound described in any one of items 35-47, or any pharmaceutically acceptable compound thereof. [Item 49] Z 3 However, C 6 -C 10 -Aryl or 5-10 member heteroaryl (1-4 heteroaryl members are independently selected from N, O, and S); n3 is either 1 or 2; R 8 However, C 1 -C 10 -It is alkyl; R 9 However, C substituted with 1 to 5 -OH groups 2 -C 6 - A compound that is alkyl, as described in item 35. [Item 50] A compound listed in item 1 or 35, or a pharmaceutically acceptable compound, selected from the table below. TIFF0007927730000270.tif231161TIFF0007927730000271.tif41162 [Item 51] A pharmaceutical composition comprising a compound described in any one of items 1 to 50 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. [Item 52] A method for selectively increasing the proliferation of cuboidal alveolar type 2 (AEC2) cells in a subject requiring such treatment, or for restoring a decrease in the proliferation of AEC2 cells in a subject requiring such treatment, comprising administering to the subject a compound described in any one of items 1 to 50 or a pharmaceutically acceptable salt thereof. [Item 53] A method for inhibiting dipeptidyl peptidase IV (DPP4) in a subject requiring such inhibition, comprising administering to the subject a compound described in any one of items 1 to 50 or a pharmaceutically acceptable salt thereof. [Item 54] A method for treating a lung disease in which a subject is suffering, comprising administering to the subject a compound described in any one of items 1 to 50 or a pharmaceutically acceptable salt thereof.

Claims

1. A compound which is a compound of formula (I) or a pharmaceutically acceptable salt thereof: 【Chemistry 1】 During the ceremony L １Ａ However, - NHCH ２ - or -CH(NH ２ ) - and; X １ is -O-, -S-, -S(O)-, S(O) ２ - and -NH- are selected; L １Ｂ wherein one or more -CH ２ - groups are independently optionally replaced by a moiety selected from -O-, -C(O)- and -NH-, and the C ２ -C １２ -alkyl; Z １ However, C ６ -C １０ - Aryls, and 5- to 10-membered heteroaryls (1 to 4 heteroaryl members independently selected from N, O, and S); m1 is an integer that is 1; n1 is an integer selected from 0, 1, 2, and 3; R １ However, H, C １ -C １０ -Alkyl and -C １ -C １０ -Alkyl-(C ６ -C １０ Selected from -aryl groups, which may be substituted with 1 to 6 -OH groups; R ２ However, C substituted with 1 to 6 -OH groups １ -C １０ - A compound that is alkyl.

2. L １Ａ ga-NHCH ２ - The compound according to claim 1.

3. L １Ａ ga-CH(NH ２ The compound according to claim 1, wherein the compound is:

4. X １ The compound according to any one of claims 1 to 3, wherein the compound is -O-.

5. Z １ C ６ -C １０ - The compound according to any one of claims 1 to 4, wherein the compound is an aryl or a 5- to 10-membered heteroaryl (where 1 to 4 heteroaryl members are independently selected from N, O, and S).

6. Z １ The compound according to any one of claims 1 to 5, wherein is phenyl.

7. Z １ The compound according to any one of claims 1 to 5, wherein is triazolyl.

8. The compound according to any one of claims 5 to 7, wherein n1 is 1 or 2.

9. R １ However, C may be substituted with 1 to 6 -OH groups. １ -C １０ - The compound according to any one of claims 5 to 8, wherein it is alkyl.

10. R １ C １ -C ６ - The compound according to any one of claims 5 to 9, wherein it is alkyl.

11. R ２ However, C is substituted with 1 to 5 -OH groups. ２ -C ６ - The compound according to any one of claims 5 to 10, wherein it is alkyl.

12. R ２ However, C is substituted with 3 to 5 -OH groups. ２ -C ６ - The compound according to any one of claims 5 to 11, wherein it is alkyl.

13. R ２ but: 【Chemistry 4】 The compound according to any one of claims 5 to 12.

14. Z １ However, C ６ -C １０ - Aryl or 5- to 10-membered heteroaryl (1 to 4 heteroaryl members independently selected from N, O, and S); n1 is 1 or 2; R １ However, C １ -C １０ - Alkyl; R ２ However, C substituted with 1 to 5 -OH groups ２ -C ６ - The compound according to claim 1, wherein it is alkyl.

15. A compound or a pharmaceutically acceptable salt thereof, selected from the table below. Table 2

16. A pharmaceutical composition comprising a compound according to any one of claims 1 to 15 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

17. A pharmaceutical composition for selectively increasing the proliferation of cuboidal alveolar type 2 (AEC2) cells in a subject requiring such proliferation, or for restoring a decrease in AEC2 cell proliferation in a subject requiring such proliferation, comprising a compound according to any one of claims 1 to 15 or a pharmaceutically acceptable salt thereof.

18. A pharmaceutical composition for inhibiting dipeptidyl peptidase IV (DPP4) in a subject requiring such inhibition, comprising a compound according to any one of claims 1 to 15 or a pharmaceutically acceptable salt thereof.

19. A pharmaceutical composition for treating a lung disease being treated, comprising a compound according to any one of claims 1 to 15 or a pharmaceutically acceptable salt thereof.

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