Antiviral compounds

Abstract

The present disclosure provides compounds for the treatment of a variety of diseases such as respiratory syncytial virus (RSV), HRV, hMPV, Ebola virus, Zika virus, West Nile virus, Dengue fever, and HCV.

Description

[0001] Cross-reference to related applications

[0002] This application claims the benefit of priority to U.S. Provisional Patent Application No. 62 / 977,881, entitled “ANTIVIRAL COMPOUNDS,” filed February 18, 2020, the entire contents of which are incorporated herein by reference.

[0003] SEQUENCE LISTING

[0004] This application includes a sequence list, which is submitted electronically in ASCII format and incorporated herein by reference in its entirety. The ASCII copy was created on February 17, 2021, and is named 1223-WO-PCT_SL.txt, with a size of 800 bytes. Background Technology

[0005] Pneumoviridae viruses are negative-sense single-stranded RNA viruses that cause many prevalent human and animal diseases. Pneumoviridae viruses include human respiratory syncytial virus (HRSV) and human metapneumovirus. Almost all children will be infected with HRSV before the age of two. HRSV is a leading cause of lower respiratory tract infections in infancy and childhood, with 0.5% to 2% of infections requiring hospitalization.

[0006] Currently, there is no vaccine to prevent HRSV infection. The monoclonal antibody palizumab can be used for immunization, but its use is limited to high-risk infants, such as premature infants or those with congenital heart or lung diseases, and its general cost is often prohibitive. Additionally, the nucleoside analog ribavirin has been approved as the only antiviral agent for treating HRSV infection, but its efficacy is limited. Therefore, antiviral treatments targeting pulmonary viruses are needed.

[0007] Examples of pyrrolo[2,3-d]pyrimidine compounds useful in the treatment of viral infections are described in U.S. 2012 / 0009147 Al (Cho et al.), U.S. 2012 / 0020921 Al (Cho et al.), WO 2008 / 089105 A2 (Babu et al.), WO 2008 / 141079 Al (Babu et al.), WO 2009 / 132135 Al (Butler et al.), WO 2010 / 002877 A2 (Francom), WO 2011 / 035231 Al (Cho et al.), WO 2011 / 035250 Al (Butler et al.), WO 2011 / 150288 Al (Cho et al.), WO 2012 / 012465 (Cho et al.), WO 2012 / 012776 Al (Mackman et al.), WO 2012 / 037038 (Clarke et al.), WO 2012 / 087596 Al (Delaney et al.), and WO 2012 / 142075 Al (Girijavallabhan et al.).

[0008] Accordingly, there is a need for compositions and methods for treating Pneumoviridae viral infections (such as HRSV infection), Flaviviridae infections (including dengue) and EBOV infection that are effective and have an acceptable toxicity profile. The present disclosure addresses the aforementioned and other needs. SUMMARY

[0009] In some embodiments, the present disclosure provides a compound of Formula (II):

[0010]

[0011] or a pharmaceutically acceptable salt thereof, wherein:

[0012] B is

[0013]

[0014] R 1A and R 2A each independently is:

[0015] (A) C1-6alkyl optionally substituted with 1 to 3 R 1B 1-12 alkyl,

[0016] (B) 3- to 6-membered heterocyclyl having 1 to 3 heteroatoms independently selected from N, O, and S, wherein the 3- to 6-membered heterocyclyl is optionally substituted with 1 to 3 R 1C or

[0017] ​(C) phenyl, wherein each R 1B is independently halogen, -OH, -NH2, C 1-6 alkyl, methoxyethoxy, C 3-8 cycloalkyl, or 3- to 6-membered heterocyclyl having 1 to 3 heteroatoms independently selected from N, O, and S, and each R 1C is independently C 1-3 alkyl;

[0018] R 3 is -N(H)R 3A or -N=C(R 3B )(R 3C ); R 3A is H, -CH2OP(O)(OH)2, or -C(O)R 3D , wherein R 3D is C 1-6 alkyl optionally substituted with 1 methoxy, or 3- to 6-membered heterocyclyl having 1 to 3 heteroatoms independently selected from N, O, and S optionally substituted with C 1-3 alkyl; R 3B is H or C 1-3 alkyl; R 3C is -N(R 3C1 )(R 3C2 ); R 3C1 and R 3C2 are each independently H or C 1-6 alkyl; or R 3C1 and R 3C2 together with the atom to which they are attached form 3- to 6-membered heterocyclyl having 1 to 3 heteroatoms independently selected from N, O, and S optionally substituted with C 1-6 alkyl;

[0019] R 4A is O or S;

[0020] R 4B and R 4C are each independently

[0021] (A) -OH;

[0022] (B) -OR 4B1 , wherein R 4B1 is C 4B2 alkyl optionally substituted with 1 to 3 R 1-6 groups, or C 6-12 aryl, wherein each R 4B2 group is independently C 1-6 alkoxy, -S-R 4B3 , or -S(O)2-R 4B3 , and each R 4B3groups are independently C 1-6 alkyl;

[0023] (C) wherein m is 0, 1, 2, 3, 4, or 5; and each R 4D is independently C 4D1 alkyl optionally substituted with 1 to 3 R 1-3 groups, or -C(O)R 4D2 is C 1-3 alkyl optionally substituted with 1 to 3 R 4D3 groups, or -C(O)N(R 4D1 )2, wherein each R 4D3 group is independently -NH2or -C(O)OR 4D2 , each R 1-3 is independently C 4D3 alkyl, and each R 1-3 is independently C

[0024] (D) wherein R 4E1 and R 4E2 are each independently H or C 1-6 alkyl, R 4F1 and R 4F2 are each independently H or C 1-6 alkyl, or R 4F1 and R 4F2 together are oxo, R 4G is C 4G1 alkyl optionally substituted with 1 to 3 R 1-12 groups, or C 4G2 cycloalkyl optionally substituted with 1 to 3 R 3-7 groups, or -C(O)R 4G3 , each R 4G4 is independently -OH, C 4G1 alkyl, C 1-6 haloalkyl, -OH, or -NH2, each R 1-3 is independently C 1-5 alkyl, C 4G8 haloalkyl, -OH, or -NH2, each R 4G9 is independently C 3-7 alkyl, C 4G10 haloalkyl, -OH, or -NH2, and each R 4G2 is independently C 1-6 alkyl, C 1-3 haloalkyl, -OH, or -NH2.4G3 independently halogen or C 1-3 alkyl, each R 4G4 independently C 1-12 alkyl, each R 4G8 independently C 1-6 alkyl, each R 4G9 independently C 1-3 haloalkyl, -OH, or -NH2, and each R 4G10 independently C 1-3 haloalkyl; or

[0025] (E)-(OP(O)(OH)) 1-2 -OH; and

[0026] R 5A and R 5B each is C 1-6 alkyl.

[0027] In another embodiment, the present disclosure provides a pharmaceutical formulation comprising a pharmaceutically effective amount of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

[0028] In another embodiment, the present disclosure provides a method of treating a Picornaviridae virus infection in a human in need thereof, comprising administering to the human a therapeutically effective amount of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof.

[0029] In another embodiment, the present disclosure provides a method of treating a Picornaviridae virus infection in a human in need thereof, comprising administering to the human a therapeutically effective amount of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof.

[0030] In another embodiment, the present disclosure provides a method of treating a Flaviviridae virus infection in a human in need thereof, comprising administering to the human a therapeutically effective amount of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof.

[0031] In another embodiment, the present disclosure provides a method of treating a Filoviridae virus infection in a human in need thereof, comprising administering to the human a therapeutically effective amount of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof.

[0032] In another embodiment, the present disclosure provides a method of making a medicament for treating a Picornaviridae virus infection in a human in need thereof, characterized in that a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, is used.

[0033] In another embodiment, the present disclosure provides a method of preparing a medicament for treating a Picornaviridae virus infection in a human in need thereof, characterized by using a compound of the present disclosure or a pharmaceutically acceptable salt thereof.

[0034] In another embodiment, the present disclosure provides a method of preparing a medicament for treating a Flaviviridae virus infection in a human in need thereof, characterized by using a compound of the present disclosure or a pharmaceutically acceptable salt thereof.

[0035] In another embodiment, the present disclosure provides a method of preparing a medicament for treating a Filoviridae virus infection in a human in need thereof, characterized by using a compound of the present disclosure or a pharmaceutically acceptable salt thereof.

[0036] In another embodiment, the present disclosure provides the use of a compound of the present disclosure or a pharmaceutically acceptable salt thereof for the preparation of a medicament for the treatment of a Pneumoviridae virus infection in a human.

[0037] In another embodiment, the present disclosure provides the use of a compound of the present disclosure or a pharmaceutically acceptable salt thereof for the preparation of a medicament for the treatment of a Picornaviridae virus infection in a human.

[0038] In another embodiment, the present disclosure provides the use of a compound of the present disclosure or a pharmaceutically acceptable salt thereof for the preparation of a medicament for the treatment of a Flaviviridae virus infection in a human.

[0039] In another embodiment, the present disclosure provides the use of a compound of the present disclosure or a pharmaceutically acceptable salt thereof for the preparation of a medicament for the treatment of a Filoviridae virus infection in a human.

[0040] In another embodiment, the present disclosure provides a compound of the present disclosure or a pharmaceutically acceptable salt thereof for use in the treatment of a Pneumoviridae virus infection in a human in need thereof.

[0041] In another embodiment, the present disclosure provides a compound of the present disclosure or a pharmaceutically acceptable salt thereof for use in the treatment of a Picornaviridae virus infection in a human in need thereof.

[0042] In another embodiment, the present disclosure provides a compound of the present disclosure or a pharmaceutically acceptable salt thereof for use in the treatment of a Flaviviridae virus infection in a human in need thereof.

[0043] In another embodiment, the present disclosure provides a compound of the present disclosure or a pharmaceutically acceptable salt thereof for use in the treatment of a Filoviridae virus infection in a human in need thereof.

[0044] In another embodiment, the present disclosure provides a method for treating or preventing exacerbation of a respiratory condition caused by a viral infection in a human in need thereof, comprising administering to said human a therapeutically effective amount of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, wherein the respiratory condition is chronic obstructive pulmonary disease.

[0045] In another embodiment, the present disclosure provides a method of manufacturing a medicament for treating or preventing exacerbation of a respiratory condition caused by a viral infection in a human in need thereof, characterized in that a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, is used, wherein the respiratory condition is chronic obstructive pulmonary disease.

[0046] In another embodiment, the present disclosure provides the use of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for treating or preventing exacerbation of a respiratory condition caused by a viral infection in a human, wherein the respiratory condition is chronic obstructive pulmonary disease.

[0047] In another embodiment, the present disclosure provides a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, for use in treating or preventing exacerbation of a respiratory condition caused by a viral infection in a human in need thereof, wherein the respiratory condition is chronic obstructive pulmonary disease. DETAILED DESCRIPTION

[0048] I. SUMMARY

[0049] The present disclosure provides 2',3'-diester-4'-cyano nucleoside compounds for treating viral infections such as Ebola, Zika, West Nile, Yellow Fever, Dengue, HCV, RSV, and the like.

[0050] II. DEFINITIONS

[0051] "Alkyl" is a straight-chain or branched saturated monovalent hydrocarbon. For example, an alkyl group can have from 1 to 18 carbon atoms (i.e., C 1-18 alkyl) or 1 to 8 carbon atoms (i.e., C 1-8 alkyl) or 1 to 6 carbon atoms (i.e., C 1-6 alkyl) or 1 to 4 carbon atoms (i.e., C 1-4Examples of alkyl groups include, but are not limited to, methyl (Me, -CH3), ethyl (Et, -CH2CH3), 1 -propyl (n-Pr, n-propyl, -CH2CH2CH3), 2-propyl (i-Pr, isopropyl, -CH(CH3)2), 1 -butyl (n-Bu, n-butyl, -CH2CH2CH2CH3), 2-methyl-l -propyl (i-Bu, isobutyl, -CH2CH(CH3)2), 2-butyl (s-Bu, sec-butyl, -CH(CH3)CH2CH3), 2-methyl-2-propyl (t-Bu, tert-butyl, -C(CH3)3), 1-pentyl (n-pentyl, -CH2CH2CH2CH2CH3), 2-pentyl (-CH(CH3)CH2CH2CH3), 3-pentyl (-CH(CH2CH3)2), 2-methyl-2-butyl (-C(CH3)2CH2CH3), 3-methyl-2-butyl (-CH(CH3)CH(CH3)2), 3-methyl-l -butyl (-CH2CH2CH(CH3)2), 2-methyl-l -butyl (-CH2CH(CH3)CH2CH3), 1-hexyl (-CH2CH2CH2CH2CH2CH3), 2-hexyl (-CH(CH3)CH2CH2CH2CH3), 3-hexyl (-CH(CH2CH3)(CH2CH2CH3)), 2-methyl-2-pentyl (-C(CH3)2CH2CH2CH3), 3-methyl-2-pentyl (-CH(CH3)CH(CH3)CH2CH3), 4-methyl-2-pentyl (-CH(CH3)CH2CH(CH3)2), 3-methyl-3-pentyl (-C(CH3)(CH2CH3)2), 2-methyl-3-pentyl (-CH(CH2CH3)CH(CH3)2), 2,3-dimethyl-2-butyl (-C(CH3)2CH(CH3)2), and 3,3-dimethyl-2-butyl (-CH(CH3)C(CH3)3). Other alkyl groups include heptyl, octyl, nonyl, decyl, undecyl, dodecyl, pentadecyl, hexadecyl, heptadecyl, and octadecyl.

[0052] “Alkoxy” refers to an alkyl group having an oxygen atom attaching the alkyl group to the point of attachment: alkyl-O-. As with alkyl groups, an alkoxy group can have any suitable number of carbon atoms, such as C 1-6 . Alkoxy groups include, for example, methoxy, ethoxy, propyloxy, isopropyloxy, butoxy, 2-butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentyloxy, hexyloxy, and the like. An alkoxy group can be further substituted with a variety of substituents described herein. An alkoxy group can be substituted or unsubstituted.

[0053] "Alkoxy-alkoxy" refers to an alkoxy group attached to a second alkoxy group, which is attached to the remainder of the compound. Alkoxy groups are as defined above and may include, but are not limited to, methoxy-methoxy (CH3OCH2O-), methoxy-ethoxy (CH3OCH2CH2O-), etc.

[0054] "Hydroxy group" refers to -OH.

[0055] As used in this article, “halogenated” or “halogen” refers to fluorine (-F), chlorine (-Cl), bromine (-Br), and iodine (-I).

[0056] As used herein, “haloalkyl” means an alkyl group as defined herein, wherein one or more hydrogen atoms of the alkyl group are independently substituted by a halogenated substituent, which may be the same or different. For example, C 1-4 The alkyl halide is C 1-4 Alkyl, wherein C 1-4 One or more hydrogen atoms in the alkyl group have been replaced by a halogenated substituent. Examples of halogenated alkyl groups include, but are not limited to, fluoromethyl, fluorochloromethyl, difluoromethyl, difluorochloromethyl, trifluoromethyl, 1,1,1-trifluoroethyl, and pentafluoroethyl.

[0057] "Cycloalkyl" refers to a compound having 3 to 20 cyclic carbon atoms (i.e., C6 ... 3-20 Cycloalkyl groups are single saturated or partially unsaturated carbon rings, such as those with 3 to 12 cyclic atoms, 3 to 10 cyclic atoms, 3 to 8 cyclic atoms, 3 to 6 cyclic atoms, 3 to 5 cyclic atoms, or 3 to 4 cyclic atoms. The term "cycloalkyl" also includes multiple fused, saturated, and partially unsaturated carbon ring systems (e.g., ring systems containing 2, 3, or 4 carbon rings). Thus, cycloalkyl groups include polycyclic carbon rings such as bicyclic carbon rings (e.g., bicyclic carbon rings having about 6 to 12 cyclic carbon atoms, such as bicyclo[3.1.0]hexane and bicyclo[2.1.1]hexane) and polycyclic carbon rings (e.g., tricyclic and tetracyclic carbon rings having up to about 20 cyclic carbon atoms). When valence requirements permit, the rings of a polyfused ring system may be linked to each other via fusion, spirocyclic, and bridging bonds. Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclohexyl, 1-cyclohex-1-enyl, 1-cyclohex-2-enyl, and 1-cyclohex-3-enyl.

[0058] As used herein, "heterocyclyl" or "heterocycle" or "heterocycloalkyl" refers to a single saturated or partially unsaturated non-aromatic ring or non-aromatic multiple ring system having at least one heteroatom (i.e., at least one ring-forming heteroatom selected from oxygen, nitrogen, and sulfur) in the ring. Unless otherwise specified, a heterocyclyl group has from 3 to about 20 ring atoms, for example, 3 to 12 ring atoms, for example, 3 to 10 ring atoms, or 3 to 8 ring atoms, or 3 to 6 ring atoms, or 3 to 5 ring atoms, or 4 to 6 ring atoms, or 4 to 5 ring atoms. Thus, the term includes single saturated or partially unsaturated rings (e.g., 3-, 4-, 5-, 6-, or 7-membered rings) having from about 1 to 6 ring carbon atoms and from about 1 to 3 ring-forming heteroatoms selected from oxygen, nitrogen, and sulfur. When valence requirements permit, the rings of a multiple fused ring (e.g., bicyclic heterocyclyl) system can be connected to one another via fused, spiro, and bridged linkages. Heterocycles include, but are not limited to, azetidine, aziridine, imidazolidine, morpholine, oxirane (epoxide), oxetane, thietane, piperazine, piperidine, pyrazolidine, piperidine, pyrrolidine, pyrrolidone, tetrahydrofuran, tetrahydrothiophene, dihydropyridine, tetrahydropyridine, quinuclidine, 2-oxa-6-azaspiro[3.3]heptan-6-yl, 6-oxa-1-azaspiro[3.3]heptan-1-yl, 2-thia-6-azaspiro[3.3]heptan-6-yl, 2,6-diazaspiro[3.3]heptan-2-yl, 2-azabicyclo[3.1.0]hexan-2-yl, 3-azabicyclo[3.1.0]hexanyl, 2-azabicyclo[2.1.1]hexanyl, 2-azabicyclo[2.2.1]heptan-2-yl, 4-azaspiro[2.4]heptanyl, 5-azaspiro[2.4]heptanyl, and the like.

[0059] As used herein, "aryl" refers to a single all-carbon aromatic ring or a multi- fused all-carbon ring system, wherein at least one ring is aromatic. For example, in some embodiments, aryl groups have 6 to 20 carbon atoms, 6 to 14 carbon atoms, or 6 to 12 carbon atoms. Aryl includes phenyl radicals. Aryl also includes multi-fused ring systems having about 9 to 20 carbon atoms (e.g., ring systems containing 2, 3, or 4 rings), wherein at least one ring is aromatic, and wherein the other rings can be aromatic or non-aromatic (i.e., carbocyclic). Such multi-fused ring systems are optionally substituted on any carbocyclic portion of the multi-fused ring system with one or more (e.g., 1, 2, or 3) oxo groups. When valence requirements permit, the rings of the multi-fused ring system can be connected to one another via fusion, spiro, and bridge bonds. It will also be appreciated that when a certain atom range is recited for an aryl group (e.g., 6-10 membered aryl), the atom range is the total ring atoms of the aryl group. For example, a 6 membered aryl would include phenyl, a 10 membered aryl would include naphthyl and 1,2,3,4-tetrahydronaphthyl. Non-limiting examples of aryl groups include, but are not limited to, phenyl, indenyl, naphthyl, 1,2,3,4-tetrahydronaphthyl, anthryl, and the like.

[0060] As used herein, "heteroaryl" refers to a monocyclic aromatic ring having at least one atom in the ring other than carbon, wherein the atom is selected from oxygen, nitrogen, and sulfur; "heteroaryl" also includes polycyclic fused ring systems having at least one such aromatic ring, which are further described below. Thus, "heteroaryl" includes a monocyclic aromatic ring having from about 1 to 6 carbon atoms and from about 1 to 4 heteroatoms selected from oxygen, nitrogen, and sulfur. The sulfur and nitrogen atoms can also be present in oxidized forms, provided that the ring is aromatic. Exemplary heteroaryl ring systems include, but are not limited to, pyridyl, pyrimidinyl, oxazolyl, or furanyl. "Heteroaryl" also includes polycyclic fused ring systems (e.g., ring systems containing 2, 3, or 4 rings), in which a heteroaryl group as defined above is fused to one or more rings selected from heteroaryl (to form, e.g., 1,8-naphthyridinyl), heterocyclo (to form, e.g., 1,2,3,4-tetrahydro-1,8-naphthyridinyl), carbocyclo (to form, e.g., 5,6,7,8-tetrahydroquinolinyl), and aryl (to form, e.g., indazolyl). Thus, a heteroaryl group (monocyclic aromatic ring or polycyclic fused ring system) has from about 1 to 20 carbon atoms and from about 1 to 6 heteroatoms within the heteroaryl ring. Such polycyclic fused ring systems can be optionally substituted on the carbocyclic or heterocyclic portion of the fused rings with one or more (e.g., 1, 2, 3, or 4) oxo groups. When valence requirements permit, the rings of the polycyclic fused ring system can be connected to one another via fused, spiro, and bridged linkages. It will be appreciated that the individual rings of the polycyclic fused ring system can be connected in any order relative to one another. It will be appreciated that the point of attachment of a heteroaryl or heteroaryl polycyclic fused ring system can be on any suitable atom of the heteroaryl or heteroaryl polycyclic fused ring system, including carbon atoms and heteroatoms (e.g., nitrogen). It will also be appreciated that when a certain atom range is recited for a heteroaryl group (e.g., 5- to 10-membered heteroaryl), the atom range is the total ring atoms of the heteroaryl group and includes both carbon atoms and heteroatoms. For example, a 5-membered heteroaryl would include thiazolyl, and a 10-membered heteroaryl would include quinolinyl. Exemplary heteroaryl groups include, but are not limited to, pyridyl, pyrrolyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrazolyl, thiophenyl, indolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, furanyl, oxadiazolyl, thiadiazolyl, quinolinyl, isoquinolinyl, benzothiazolyl, benzoxazolyl, indazolyl, quinoxalinyl, quinazolinyl, 5,6,7,8-tetrahydroisoquinolinyl, benzofuranyl, benzoimidazolyl, thienyl, pyrrolo[2,3-b]pyridyl, quinazolin-4(3H)-one, and triazolyl.

[0061] "Compounds of the disclosure" include the compounds disclosed herein, for example, the compounds of the disclosure include compounds of Formulae (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), and (IIn), including the compounds of the Examples.

[0062] “Pharmaceutically effective amount” refers to the amount of a compound of the present disclosure in a formulation or combination thereof that provides the desired therapeutic or pharmaceutical result.

[0063] “Pharmaceutically acceptable excipient” includes, but is not limited to, any auxiliary, carrier, excipient, glidant, sweetening, diluting, preservative, dye / colorant, flavor enhancer, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent or emulsor that has been approved by the U.S. Food and Drug Administration as acceptable for use in humans or domestic animals.

[0064] “Treatment” or “treat” or “treating” as used herein refers to an approach for obtaining beneficial or desired results. For purposes of the present disclosure, beneficial or desired results include, but are not limited to, reduction and / or alleviation of symptoms, and / or signs of the disease or condition associated with the disease or condition. In one embodiment, “treatment” or “treating” includes one or more of the following: (a) inhibiting the disease or condition (e.g., reducing one or more symptoms resulting from the disease or condition, and / or diminishing the extent of the disease or condition); (b) slowing or arresting the development of one or more symptoms associated with the disease or condition (e.g., stabilizing the disease or condition, delaying the worsening or progression of the disease or condition); and (c) relieving the disease or condition, e.g., causing regression of clinical symptoms, improving the disease state, delaying progression of the disease, improving quality of life, and / or prolonging survival.

[0065] “Prevention” refers to preventing or delaying the progression of clinical disease in a patient having a viral infection.

[0066] “Respiratory condition” refers to a disease or condition such as a respiratory infection caused by a viral infection, allergic rhinitis, nasal congestion, rhinorrhea, perennial rhinitis, rhinitis, all types of asthma, chronic obstructive pulmonary disease (COPD), chronic or acute bronchoconstriction, chronic bronchitis, small airway obstruction, emphysema, chronic eosinophilic pneumonia, adult respiratory distress syndrome, exacerbation of airway hyperresponsiveness due to other drug therapies, pulmonary vascular disease (including pulmonary hypertension), acute lung injury, bronchiectasis, sinusitis, allergic conjunctivitis, idiopathic pulmonary fibrosis, or atopic dermatitis, in particular asthma or allergic rhinitis or atopic dermatitis or allergic conjunctivitis.

[0067] “Respiratory condition exacerbation” refers to an exacerbation induced by a viral infection. Representative viral infections include, but are not limited to, respiratory syncytial virus (RSV), rhinovirus, and metapneumovirus.

[0068] As used herein, “therapeutically effective amount” or “effective amount” refers to an amount that is effective to elicit the desired biological or medical response, including the amount of a compound that, when administered to a subject for treatment of a disease, is sufficient to effect such treatment of the disease. The effective amount will vary depending on the compound, the disease and its severity, and the age, weight, etc. of the subject to be treated. An effective amount can include a range of amounts. As is understood in the art, an effective amount can be one or more doses, i.e., a single dose or multiple doses can be required to achieve the desired therapeutic endpoint. An effective amount can be considered in the context of administering one or more therapeutic agents, and a single agent can be considered to be given in an effective amount if, in conjunction with one or more other agents, a desired or beneficial result is achieved or realized. Suitable dosages of any co-administered compounds can optionally be reduced due to the combined effect (e.g., additive or synergistic effect) of the compounds.

[0069] As used herein, “co-administration” refers to administration of a unit dose of a compound disclosed herein prior to or following administration of a unit dose of one or more additional therapeutic agents, e.g., administration of a compound disclosed herein within seconds, minutes, or hours of administration of one or more additional therapeutic agents. For example, in some embodiments, a unit dose of a compound of the disclosure is administered first, followed by administration of a unit dose of one or more additional therapeutic agents within seconds or minutes. Alternatively, in other embodiments, a unit dose of one or more additional therapeutic agents is administered first, followed by administration of a unit dose of a compound of the disclosure within seconds or minutes. In some embodiments, a unit dose of a compound of the disclosure is administered first, followed by administration of a unit dose of one or more additional therapeutic agents after several hours (e.g., 1-12 hours). In other embodiments, a unit dose of one or more additional therapeutic agents is administered first, followed by administration of a unit dose of a compound of the disclosure after several hours (e.g., 1-12 hours). Co-administration of a compound disclosed herein and one or more additional therapeutic agents generally refers to concurrent or sequential administration of a compound disclosed herein and one or more additional therapeutic agents such that a therapeutically effective amount of each agent is present in the patient.

[0070] Also provided are pharmaceutically acceptable salts, hydrates, solvates, tautomeric forms, polymorphs, and prodrugs of the compounds described herein. “Pharmaceutically acceptable” or “physiologically acceptable” refers to compounds, salts, compositions, dosage forms, and other materials that are useful in preparing a pharmaceutical composition suitable for use in a suitable veterinary or human pharmaceutical use.

[0071] The compounds described herein can be prepared and / or formulated as pharmaceutically acceptable salts, or, where appropriate, as free bases. Pharmaceutically acceptable salts are non-toxic salts of the free bases forms of the compounds, which salts possess the desired pharmacological activity of the free bases. These salts can be derived from inorganic or organic acids. For example, compounds containing a basic nitrogen can be prepared as pharmaceutically acceptable salts by contacting the compound with an inorganic or organic acid. Non-limiting examples of pharmaceutically acceptable salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, hexanoate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, butyne-1,4-dioate, hexyne-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, sulfonate, methylsulfonate, propylsulfonate, benzenesulfonate, xylene sulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, gamma-hydroxybutyrate, glycolate, tartarate, and mandelate. Lists of other suitable pharmaceutically acceptable salts are found in "Remington: The Science and Practice of Pharmacy", 21stEdition, Lippincott Wiliams and Wilkins, Philadelphia, Pa., 2006.

[0072] Examples of "pharmaceutically acceptable salts" of the compounds disclosed herein also include salts derived from the appropriate bases such as alkali metals (for example, sodium, potassium), alkaline earth metals (for example, magnesium), ammonium and NX4 + (wherein X is C1-C4alkyl). Also included are base addition salts such as sodium or potassium salts.

[0073] Also provided are compounds described herein, or pharmaceutically acceptable salts, isomers, or mixtures thereof, wherein from 1 to n hydrogen atoms attached to a carbon atom can be replaced with deuterium atoms or D, wherein n is the number of hydrogen atoms in the molecule. As known in the art, a deuterium atom is a non-radioactive isotope of a hydrogen atom. Such compounds can increase resistance to metabolism, and thus can be useful for increasing the half-life of a compound described herein, or a pharmaceutically acceptable salt, isomer, or mixture thereof, when administered to a mammal. See, e.g., Foster, "Deuterium Isotope Effects in Studies of Drug Metabolism," Trends Pharmacol. Sci., 5(12):524-527 (1984). Such compounds are synthesized by methods well known in the art, e.g., by employing starting materials in which one or more hydrogen atoms have been replaced by deuterium.

[0074] Examples of isotopes that can be incorporated into the disclosed compounds include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, chlorine, and iodine, such as 2 H, 3 H, 11 C, 13 C, 14 C, 13 N, 15 N, 15 O, 17 O, 18 O, 31 P, 32 P, 35 S, 18 F, 36 Cl, 123 I, and 125 I. The substitution of positron emitting isotopes, such as 11 C, 18 F, 15 O, and 13 N, are useful in positron emission tomography (PET) studies to examine substrate receptor occupancy. Isotopically-labeled compounds of Formula (I) can generally be prepared by conventional techniques known to those skilled in the art, or by processes analogous to those described below, using an appropriate isotopically-labeled reagent in place of the non-labeled reagent previously employed.

[0075] The compounds of the embodiments disclosed herein, or their pharmaceutically acceptable salts, may include one or more asymmetric centers, and thus may produce enantiomers, diastereomers, and other stereoisomers that may be defined by absolute stereochemistry as (R)- or (S)- or for amino acids as (D)- or (L)-. This disclosure is intended to include all such possible isomers as well as their racemic and optically pure forms. Optically active (+) and (-), (R)- and (S)- or (D)- and (L)- isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques such as chromatography and fractional crystallization. Conventional techniques for preparing / separating individual enantiomers include chiral synthesis from suitable optically pure precursors or resolution of racemates (or racemates of salts or derivatives) using, for example, chiral high-performance liquid chromatography (HPLC). When the compounds described herein contain an alkene double bond or other geometrically asymmetric centers, and unless otherwise stated, the compounds are intended to include E and Z geometric isomers. Similarly, all tautomers are also intended to be included. When a compound is represented in its chiral form, it should be understood that the embodiments cover, but are not limited to, specific diastereomeric or enantiomerically enriched forms. When chirality is not specified but is present, it should be understood that the embodiments relate to specific diastereomeric or enantiomerically enriched forms; or racemic or non-racemic mixtures of such compounds. As used herein, a “non-racemic mixture” is a mixture of stereoisomers in a ratio not equal to 1:1.

[0076] A "racemate" is a mixture of enantiomers. The mixture may contain equal or unequal amounts of each enantiomer.

[0077] One or more “stereoisomers” refer to compounds with different chiralities of one or more stereocenters. Stereoisomers include enantiomers and diastereomers. If a compound has one or more asymmetric centers or has asymmetric substituted double bonds, the compound may exist in stereoisomeric form and can therefore be prepared as individual stereoisomers or as mixtures. Unless otherwise specified, this specification is intended to include individual stereoisomers as well as mixtures. Methods for determining stereochemistry and isolating stereoisomers are well known in the art (see, for example, Chapter 4 of Advanced Organic Chemistry, 4th Edition, J. March, John Wiley and Sons, New York, 1992).

[0078] "Tautomers" refer to alternative forms of compounds with different proton positions, such as enol-ketone and imine-enamine tautomers, or tautomers containing heteroaryl groups that are simultaneously attached to ring atoms of cyclic -NH- and cyclic =N-, such as pyrazoles, imidazoles, benzimidazoles, triazoles, and tetraazoles.

[0079] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Colons are used at the end of chemical groups for convenience; chemical groups can be depicted without one or more colons without loss of their ordinary meaning. A wavy line drawn by a line in a structure indicates a point of attachment of a group. Dotted lines indicate optional bonds. Unless chemically or structurally necessary, the order of writing of chemical groups or their point of attachment to the rest of the molecule does not indicate or imply directionality. For example, the group “-SO2CH2-” is equivalent to “-CH2SO2-” and both can be attached in either direction. Similarly, an “arylalkyl” group, for example, can be attached to the rest of the molecule at either the aryl or alkyl portion of the group. Prefixes such as “C u-v ” or (C u -C v ) indicate that the following group has u to v carbon atoms. For example, “C 1-6 alkyl” and “C1-C6alkyl” both indicate that the alkyl group has 1 to 6 carbon atoms.

[0080] As used herein, “solvate” refers to an interaction of a solvent and a compound. Solvates of the salts of the compounds described herein are also provided. Hydrates of the compounds described herein are also provided.

[0081] As used herein, “prodrug” is a derivative of a drug that is converted into the parent drug, according to some chemical or enzymatic pathway, when introduced into the body.

[0082] III. COMPOUNDS

[0083] The present disclosure provides compounds of Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), and (IIn).

[0084] In some embodiments, the present disclosure provides a compound of Formula (II):

[0085]

[0086] or a pharmaceutically acceptable salt thereof, wherein:

[0087] Base is

[0088]

[0089] R 1A and R 2A each independently is:

[0090] (A) is optionally substituted with 1 to 3 R1B substituted C 1-12 alkyl,

[0091] (B) 3- to 6-membered heterocyclyl having 1 to 3 heteroatoms independently selected from N, O, and S, wherein the 3- to 6-membered heterocyclyl is optionally substituted with 1 to 3 R 1C substituted, or

[0092] (C) phenyl, wherein each R 1B is independently halogen, -OH, -NH2, C 1-6 alkoxy, methoxyethoxy, C 3-8 cycloalkyl, or 3- to 6-membered heterocyclyl having 1 to 3 heteroatoms independently selected from N, O, and S, and each R 1C is independently C 1-3 alkyl;

[0093] R 3 is -N(H)R 3A or -N=C(R 3B )(R 3C ); R 3A is H, -CH2OP(O)(OH)2, or -C(O)R 3D , wherein R 3D is C 1-6 alkyl optionally substituted with 1 methoxy, or 3- to 6-membered heterocyclyl having 1 to 3 heteroatoms independently selected from N, O, and S optionally substituted with C 1-3 alkyl; R 3B is H or C 1-3 alkyl; R 3C is -N(R 3C1 )(R 3C2 ); R 3C1 and R 3C2 are each independently H or C 1-6 alkyl; or R 3C1 and R 3C2 together with the atom to which they are attached form 3- to 6-membered heterocyclyl having 1 to 3 heteroatoms independently selected from N, O, and S optionally substituted with C 1-6 alkyl;

[0094] R 4A is O or S;

[0095] R 4B and R 4C are each independently

[0096] (A) -OH;

[0097] (B) -OR 4B1 , wherein R 4B1 is optionally substituted with 1 to 3 R4B2 C1-C6alkyl substituted with 0, 1, 2, or 3 R 1-6 C1-C6alkyl, or C 6-12 aryl, wherein each R 4B2 group is independently C 1-6 alkoxy, -S-R 4B3 , or -S(O)2-R 4B3 , and each R 4B3 group is independently C 1-6 alkyl;

[0098] (C) wherein m is 0, 1, 2, 3, 4, or 5; and each R 4D is independently C 4D1 alkyl substituted with 0, 1, 2, or 3 R 1-3 group is independently C 4D2 alkyl substituted with 0, 1, 2, or 3 R 1-3 alkoxy, or -C(O)N(R 4D3 )2, wherein each R 4D1 group is independently -NH2or -C(O)OR 4D3 , each R 4D2 is independently C 1-3 alkyl, and each R 4D3 is independently C 1-3 alkyl;

[0099] (D) wherein R 4E1 and R 4E2 are each independently H or C 1-6 alkyl, R 4F1 and R 4F2 are each independently H or C 1-6 alkyl, or R 4F1 and R 4F2 together are oxo, R 4G is C 4G1 alkyl optionally substituted with 1 to 3 R 1-12 groups, C 4G2 cycloalkyl optionally substituted with 1 to 3 R 3-7 groups, a 3- to 8-membered heterocyclyl having 1 to 3 heteroatoms selected from N, O, and S optionally substituted with 1 to 3 R 4G3 groups, or -C(O)R 4G4 , each R 4G1 is independently -OH, C 1-6 alkyl, C 1-3 alkoxy, -(CH2OCH2) 1-5 -CH3, -N(R 4G8 )2, -OP(O)(OH)2, optionally substituted with 1 to 3 R 4G9substituted C 3-7 cycloalkyl, optionally substituted with 1 to 3 R 4G10 substituted 3- to 6-membered heterocyclyl having 1 to 3 heteroatoms independently selected from N, O, and S, or phenyl, each R 4G2 independently C 1-6 alkyl, C 1-3 haloalkyl, -OH, or -NH2, each R 4G3 independently halo or C 1-3 alkyl, each R 4G4 independently C 1-12 alkyl, each R 4G8 independently C 1-6 alkyl, each R 4G9 independently C 1-3 haloalkyl, -OH, or -NH2, and each R 4G10 independently C 1-3 haloalkyl; or

[0100] (E)-(OP(O)(OH)) 1-2 -OH; and

[0101] R 5A and R 5B each is C 1-6 alkyl.

[0102] In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein Base is

[0103]

[0104] In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein Base is

[0105]

[0106] In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein Base is

[0107]

[0108] In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein Base is

[0109]

[0110] In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2A each independently is: (A) C 1B alkyl optionally substituted with 1 to 3 R 1-12 , (B) 3- to 6-membered heterocyclyl having 1 to 3 heteroatoms independently selected from N, O, and S, wherein the 3- to 6-membered heterocyclyl is optionally substituted with 1 to 3 R 1C , or (C) phenyl, wherein each R 1B is independently -OH, -NH2, C 1-6 alkoxy, methoxyethoxy, C 3-8 cycloalkyl, or 3- to 6-membered heterocyclyl having 1 to 3 heteroatoms independently selected from N, O, and S, and each R 1C is independently C 1-3 alkyl.

[0111] In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2A is C 1B alkyl optionally substituted with 1 to 3 R 1-12alkyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2A each is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, and isoamyl, each optionally substituted with 1 to 3 R 1B In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein each R 1B may independently be methoxy, methoxyethoxy, morpholino, -OH, or -NH2. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2A each independently is methyl optionally substituted with methoxy, methoxyethoxy, or morpholino, ethyl optionally substituted with methoxy, n-propyl, isopropyl, n-butyl, isobutyl optionally substituted with -OH or -NH2, t-butyl, or isoamyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2A each independently is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, or isoamyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2Aeach independently is methyl, ethyl, or isopropyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 1A each is methyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 2A each is methyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 1A each is methyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 2A each is ethyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 1A each is ethyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 2A each is isopropyl.

[0112] In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 1A each is methyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 2A each is a 3- to 6-membered heterocyclyl having 1 to 3 heteroatoms independently selected from N, O, and S, wherein the 3- to 6-membered heterocyclyl is optionally substituted with 1 to 3 R 1C each R 1C is independently C 1-3 alkyl, hydroxyl, or halogen. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 1A each is methyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 2A each is a 4- to 6-membered heterocyclyl having 1 heteroatom selected from N and O, wherein the 4- to 6-membered heterocyclyl is optionally substituted with 1 R 1C each R 1C is independently C 1-3Alkyl group. In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2A Each is chosen by 1 R. 1C Substituted oxocyclic butyl, tetrahydropyranyl, or piperidinyl, wherein each R 1C Independently for C 1-3 Alkyl group. In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein each R 1C The compound is methyl. In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2A Each is independently an oxetyl, tetrahydropyranyl, or piperidinyl group optionally substituted with a methyl group.

[0113] In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2A Each is a phenyl group.

[0114] In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein R 3 -N(H)R 3A Or -N = C(R) 3B (R) 3C ), R 3A It can be H, -CH2OP(O)(OH)2 or -C(O)R 3D , where R 3DC, optionally substituted with one methoxy group 1-6 Alkyl, or optionally C 1-3 Alkyl-substituted 3- to 6-membered heterocyclic groups having 1 to 3 heteroatoms independently selected from N, O, and S, R 3B For H or C 1-3 Alkyl, R 3C -N(R) 3C1 (R) 3C2 ), R 3C1 and R 3C2 Each independently is H or C 1-6 Alkyl, or R 3C1 and R 3C2 Together with the atoms they are attached to, they form optionally C 1-6 Alkyl-substituted 3- to 6-membered heterocyclic groups having 1 to 3 heteroatoms independently selected from N, O, and S. In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein R 3 -N(H)R 3A And R 3A It can be H, -CH2OP(O)(OH)2 or -C(O)R 3D , where R 3D C, optionally substituted with one methoxy group 1-6 Alkyl, or optionally C 1-3 Alkyl-substituted 3- to 6-membered heterocyclic groups having 1 to 3 heteroatoms independently selected from N, O, and S. In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein R 3 -N(H)R 3A And R 3A It can be H, CH2OP(O)(OH)2 or -C(O)R 3D , where R 3D C, optionally substituted with one methoxy group 1-3 Alkyl groups, or optionally piperidines substituted with methyl groups. In some embodiments, the compound may be represented by formulas (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein R3A H. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 3 -NH2. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 3 -N=C(R3B)(R3C), wherein R 3B is H or methyl, R 3C is -N(R 3C1 )(R 3C2 ), R 3C1 and R 3C2 are each independently H or methyl, or R 3C1 and R 3C2 together with the atoms to which they are attached form piperazine optionally substituted with methyl.

[0115] In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4A is O or S. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4A is O. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4A is S.

[0116] In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein R 4B and R 4C One of them is -OH.

[0117] In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein R 4B and R 4C One of them is –OR 4B1 , where R 4B1 To be arbitrarily selected by 1 to 3 R 4B2 C with substituent group 1-6 Alkyl, or C 6-12 Aryl, where each R 4B2 The group is independently C 1-6 Alkoxy, -SR 4B3 or -S(O)2-R 4B3 And each R 4B3 The group is independently C 1-6 Alkyl group. In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein R 4B1 To be arbitrarily selected by 1 to 3 R 4B2 C with substituent group 1-6 Alkyl group. In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein each R 4B2 The group is independently C 1-6 Alkoxy, -SR 4B3 or -S(O)2-R 4B3 And each R 4B3 The group is independently C 1-6Alkyl group. In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein R 4B1 To be arbitrarily selected by 1 to 3 R 4B2 C with substituent group 1-6 Alkyl, wherein each R 4B2 The group is independently C 1-6 Alkoxy, -SR 4B3 or -S(O)2-R 4B3 And each R 4B3 The group is methyl. In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm) or (IIIn) or a pharmaceutically acceptable salt thereof, wherein R 4B and R 4C One of them is -OC, which is optionally substituted with methoxy, methylthio, or methanesulfonyl groups. 1-6 alkyl.

[0118] In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein R 4B and R 4C One of them is

[0119]

[0120] Where m is 0, 1, 2, 3, 4, or 5; and each R 4D Independently, it can be optionally divided into 1 to 3 Rs. 4D1 C with substituent group 1-3 Alkyl groups, optionally with 1 to 3 R groups 4D2 C with substituent group 1-3 alkoxy group, or -C(O)N(R) 4D3 )2, where each R 4D1 The functional group is independently -NH2 or -C(O)OR 4D3 Each R 4D2 Independently for C 1-3 Alkoxy groups, and each R 4D3 Independently for C 1-3 alkyl.

[0121] In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein m is 0, 1, or 2; and each R 4D is independently C 4D1 alkyl optionally substituted with 1 R 1-3 alkyl, C 1-3 alkoxy optionally substituted with methoxy, or -C(O)N(R 4D3 )2, wherein each R 4D1 group is independently -NH2or -C(O)OR 4D3 , and each R 4D3 is independently C 1-3 alkyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein m is 0 or 1. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein m is 0.

[0122] In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4B and R 4C are each independently H or C

[0123]

[0124] wherein R 4E1 and R 4E2 are each independently H or C 1-6 alkyl, R 4F1 and R 4F2 are each independently H or C 1-6 alkyl, or R 4F1 and R 4F2 together are oxo, R 4G is optionally substituted with 1 to 3 R 4G1substituted C 1-12 alkyl, optionally substituted with 1 to 3 R 4G2 substituted C 3-7 cycloalkyl, optionally substituted with 1 to 3 R 4G3 substituted 3- to 8-membered heterocyclyl having 1 to 3 heteroatoms selected from N, O, and S, or -C(O)R 4G4 each R 4G1 independently -OH, C 1-6 alkyl, C 1-3 alkoxy, -(CH2OCH2) 1-5 -CH3, -N(R 4G8 )2, -OP(O)(OH)2, optionally substituted with 1 to 3 R 4G9 substituted C 3-7 cycloalkyl, optionally substituted with 1 to 3 R 4G10 substituted 3- to 6-membered heterocyclyl having 1 to 3 heteroatoms independently selected from N, O, and S, or phenyl, each R 4G2 independently C 1-6 alkyl, C 1-3 haloalkyl, -OH, or -NH2, each R 4G3 independently halo or C 1-3 alkyl, each R 4G4 independently C 1-12 alkyl, each R 4G8 independently C 1-6 alkyl, each R 4G9 independently C 1-3 haloalkyl or -NH2; and each R 4G10 independently C 1-3 haloalkyl.

[0125] In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4B one of R 4C and R

[0126] and

[0127] R 4B and R 4C the other of R

[0128]

[0129] In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4E1 and R 4E2 are each independently H or C 1-6 alkyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4E1 and R 4E2 are each independently H or methyl.

[0130] In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4F1 and R 4F2 are each independently H or C 1-6 alkyl, or R 4F1 and R 4F2 are taken together as oxo. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4F1 and R 4F2 are each independently H or methyl, or R 4F1 and R 4F2 are taken together as oxo. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4F1 and R 4F2 are taken together as oxo.

[0131] In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein R 4B and R 4C One of them is:

[0132]

[0133] In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein R 4B and R 4C One of them is:

[0134]

[0135] In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein R 4G To be arbitrarily selected by 1 to 3 R 4G1 Replacement C 1-12 Alkyl group. In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein R 4G Each can be arbitrarily assigned to 1 to 3 Rs 4G1 The substituted methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, or 2-ethylbutyl.

[0136] In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein each R 4G1 Independently -OH, C 1-6 Alkyl, C 1-3 Alkyl group, -(CH2OCH2)1-5 -CH3, -N(R 4G8 )2, -OP(O)(OH)2, C 4G9 alkyl optionally substituted with 1 to 3 R 3-7 cycloalkyl optionally substituted with 1 to 3 R 4G10 heterocyclyl having 3 to 6 members independently selected from N, O, and S, or phenyl, wherein each R 4G8 independently is C 1-6 alkyl, each R 4G9 independently is C 1-3 haloalkyl or -NH2, and each R 4G10 independently is C 1-3 alkyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein each R 4G1 independently is -OH, C 1-6 alkyl, C 1-3 alkoxy, -(CH2OCH2) 1-5 -CH3, -N(R 4G8 )2, -OP(O)(OH)2, C 4G9 alkyl optionally substituted with 1 to 3 R 4-6 cycloalkyl optionally substituted with 1 to 3 R 4G10 heterocyclyl having 4 to 6 members independently selected from N and O, or phenyl, wherein each R 4G8 independently is C 1-6 alkyl, each R 4G9 independently is C 1-3 haloalkyl or -NH2, and each R 4G10 independently is C 1-3 haloalkyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein each R 4G1 independently is -OH, methyl, OMe, -(CH2OCH2)2-CH3, -N(iPr)2, -OP(O)(OH)2, cyclopropyl, cyclobutyl, cyclohexyl optionally substituted with -NH2or CF3, oxetanyl, piperidinyl optionally substituted with CF3or CH2CF3, tetrahydropyranyl, morpholinyl, or phenyl.

[0137] In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4G is methyl optionally substituted with R 4G1 is ethyl optionally substituted with morpholinyl or -N(R 4G8 )2, n-propyl optionally substituted with methoxy or morpholinyl, isopropyl, n-butyl optionally substituted with C 1-3 alkyl, isobutyl optionally substituted with -OH or -OP(O)(OH)2, wherein R 4G1 is cyclopropyl, cyclobutyl, cyclohexyl optionally substituted with R 4G9 , oxetanyl, piperidinyl optionally substituted with R 4G10 , tetrahydropyranyl, or phenyl.

[0138] In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4G8 is C 1-3 alkyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4G8 is isopropyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4G9 is C 1-3 haloalkyl or -NH2. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4G9-CF3or -NH2. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4G10 C 1-3 haloalkyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4G10 -CF3or 2,2,2-trifluoroethyl.

[0139] In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4G methyl optionally substituted with R 4G1 ethyl optionally substituted with morpholinyl or -N(C 1-3 alkyl)2, n-propyl optionally substituted with methoxy or morpholinyl, isopropyl, n-butyl optionally substituted with C 1-3 alkyl, isobutyl optionally substituted with -OH or -OP(O)(OH)2, wherein R 4G1 cyclopropyl, cyclobutyl, piperidinyl optionally substituted with -NH2or C 1-3 haloalkyl, oxetanyl, tetrahydropyranyl optionally substituted with C 1-3 alkyl or C 1-3 haloalkyl, piperidinyl, tetrahydropyranyl, or phenyl.

[0140] In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4G C 4G2 optionally substituted with 1 to 3 R 3-7cycloalkyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4G is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, each of which is optionally substituted with 1 to 3 R 4G2 In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein each R 4G2 is independently C 1-6 alkyl, C 1-3 haloalkyl, or -NH2. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein each R 4G2 is t-butyl, -CF3, or -NH2.

[0141] In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4G is a 3- to 8-membered heterocyclyl having 1 to 3 heteroatoms selected from N, O, and S, optionally substituted with 1 to 3 R 4G3 In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4G is a 3- to 6-membered heterocyclyl having 1 to 3 heteroatoms selected from N, O, and S, optionally substituted with 1 to 3 R 4G3 In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R4G To be arbitrarily selected by 1 to 3 R 4G3 The substituted compound is a 4- to 6-membered heterocyclic group having one to two heteroatoms selected from N and O. In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein R 4G Each can be arbitrarily assigned to 1 to 3 Rs 4G3 Substituted oxetane butyl, pyrrolidinyl, piperidinyl, tetrahydrofuranyl, or tetrahydropyranyl. In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein each R 4G3 Independently halogen or C 1-3 Alkyl group. In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein each R 4G3 It can be F, methyl, or ethyl independently.

[0142] In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein R 4G -C(O)R 4G4 In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein each R 4G4 Independently for C 1-12 Alkyl group. In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein R 4G-C(O)C 1-6 alkyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4G -C(O)-tert-butyl.

[0143] In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4B and one of R 4C is -(OP(O)(OH)) 1-2 -OH. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4B and one of R 4C is -(OP(O)(OH)) 1-2 -OH, and R 4B and one of R 4C is -OH.

[0144] In some embodiments, the compound can be represented by Formula (II), (IIa), or (IIb), or a pharmaceutically acceptable salt thereof, wherein R 5A and R 5B are each -CH2OP(O)(OH)2. In some embodiments, the compound can be represented by Formula (II) or (IIa), wherein R 5A is -CH2OP(O)(OH)2. In some embodiments, the compound can be represented by Formula (II) or (IIb), or a pharmaceutically acceptable salt thereof, wherein R 5B is -CH2OP(O)(OH)2.

[0145] In some embodiments, the compound of the present disclosure is represented by Formula (IIa), or a pharmaceutically acceptable salt thereof:

[0146]

[0147] wherein R 5A is -CH2OP(O)(OH)2.

[0148] In some embodiments, the compounds of the disclosure are represented by Formula (IIb), or a pharmaceutically acceptable salt thereof:

[0149]

[0150] wherein R 5B is -CH2OP(O)(OH)2.

[0151] In some embodiments, the compounds can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein

[0152] R 1A and R 2A are each independently methyl optionally substituted with methoxy, methoxyethoxy, or morpholinyl, ethyl optionally substituted with methoxy, n-propyl, isopropyl, n-butyl, isobutyl optionally substituted with -OH or -NH2, t-butyl, isoamyl, oxetanyl, tetrahydropyranyl, piperidinyl optionally substituted with methyl, or phenyl;

[0153] R 3 is -N(H)R 3A or -N=C(R 3B )(R 3C ); R 3A is H, -C(H)2OP(O)(OH)2, or -C(O)R 3D ; R 3D is methyl, ethyl optionally substituted with methoxy, isopropyl, or piperidinyl optionally substituted with methyl; R 3B is H or methyl; R 3C is -N(R 3C1 )(R 3C2 ); R 3C1 and R 3C2 are independently H or methyl; or R 3C1 and R 3C2 together with the atoms to which they are attached form piperazinyl optionally substituted with methyl;

[0154] R 4A is O or S; and

[0155] R 4B and R 4C are each independently

[0156] (A) -OH;

[0157] (B) -O-C optionally substituted with methoxy, methylthio, or methylsulfonyl 1-6 alkyl;

[0158] (C)

[0159] wherein m is 0, 1, or 2; and each R 4D is independently C 4D1 alkyl optionally substituted with 1 R 1-3 group, C 1-3 alkoxy optionally substituted with methoxy, or -C(O)N(R 4D3 )2, wherein each R 4D1 group is independently -NH2or -C(O)OR 4D3 , and each R 4D3 is independently C 1-3 alkyl; or

[0160] (D)

[0161] wherein R 4E1 and R 4E2 are each independently H or methyl, R 4F1 and R 4F2 are each independently H or methyl, or R 4F1 and R 4F2 together are oxo, R 4G is methyl optionally substituted with R 4G1 , ethyl optionally substituted with morpholinyl or -N(C 1-3 alkyl)2, n-propyl optionally substituted with methoxy or morpholinyl, isopropyl, n-butyl optionally substituted with C 1-3 alkyl, isobutyl optionally substituted with -OH or -OP(O)(OH)2, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl optionally substituted with 1 to 3 groups each independently -NH2, C 1-6 alkyl, or C 1-3 haloalkyl, oxetanyl, pyrrolidinyl optionally substituted with 1 to 3 methyl groups, piperidinyl optionally substituted with halogen or C 1-3 alkyl, tetrahydrofuranyl, tetrahydropyranyl, or -C(O)C 1-6 alkyl, and R 4G1 is cyclopropyl, cyclobutyl, cyclohexyl optionally substituted with -NH2or C 1-3 haloalkyl, oxetanyl, piperidinyl optionally substituted with C 1-3 haloalkyl, tetrahydropyranyl, or phenyl.

[0162] In some embodiments, the compounds of the present disclosure can be represented by formula (IIc), or a pharmaceutically acceptable salt thereof:

[0163]

[0164] In some embodiments, the compounds of the disclosure can be represented by Formula (IId), or a pharmaceutically acceptable salt thereof:

[0165]

[0166] In some embodiments, the compounds of the disclosure can be represented by Formula (IId), or a pharmaceutically acceptable salt thereof:

[0167]

[0168] In some embodiments, the compounds of the disclosure can be represented by Formula (IIf), or a pharmaceutically acceptable salt thereof:

[0169]

[0170]

[0171] In some embodiments, the compounds of the disclosure can be represented by Formula (IIg), or a pharmaceutically acceptable salt thereof:

[0172]

[0173] wherein m is 0 or 1.

[0174] In some embodiments, the compounds of the disclosure can be represented by Formula (IIh), or a pharmaceutically acceptable salt thereof:

[0175]

[0176] In some embodiments, the compounds of the disclosure can be represented by Formula (IIi), or a pharmaceutically acceptable salt thereof:

[0177]

[0178] In some embodiments, the compounds of the disclosure can be represented by Formula (IIj), or a pharmaceutically acceptable salt thereof:

[0179]

[0180] In some embodiments, the compounds of the disclosure can be represented by Formula (IIk), or a pharmaceutically acceptable salt thereof:

[0181]

[0182] In some embodiments, the compounds of the disclosure can be represented by Formula (IIm), or a pharmaceutically acceptable salt thereof:

[0183]

[0184] In some embodiments, the compounds of the disclosure can be represented by Formula (IIn), or a pharmaceutically acceptable salt thereof:

[0185]

[0186] In some embodiments, the compounds can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2A each independently is methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, or i-pentyl. In some embodiments, the compounds can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2A each independently is methyl, ethyl, or i-propyl. In some embodiments, the compounds can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2A each independently is methyl. In some embodiments, the compounds can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2A each independently is ethyl. In some embodiments, the compounds can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2A each independently is i-propyl.

[0187] In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4G is methyl, ethyl, n-propyl, i-propyl, n-butyl, t-butyl, or 2-ethylbutyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4G is methyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4G is ethyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4G is n-propyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4G is i-propyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4G is n-butyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 4GIt is tert-butyl. In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm) or (IIIn) or a pharmaceutically acceptable salt thereof, wherein R 4G It is 2-ethylbutyl.

[0188] In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2A Each of the following is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, or isopentyl, R 4G For optional use by R 4G1 Substituted methyl, optionally morpholino, or -N(C 1-3 alkyl)2-substituted ethyl, optionally methoxy or morpholino-substituted n-propyl, isopropyl, optionally C 1-3 Alkyl-substituted n-butyl, isobutyl optionally substituted with -OH or -OP(O)(OH)2, cyclopropyl, cyclobutyl, cyclopentyl, optionally substituted with 1 to 3 independently -NH2, C 1-6 Alkyl or C 1-3 The alkyl group substituted with a cyclohexyl group, oxetyl group, pyrroleyl group optionally substituted with 1 to 3 methyl groups, or alkyl group optionally substituted with a halogen or C group 1-3 Alkyl-substituted piperidinyl, tetrahydrofuranyl, tetrahydropyranyl, or -C(O)C 1-6 Alkyl, and R 4G1 Cyclopropyl, cyclobutyl, optionally with -NH2 or C 1-3 Halogenated alkyl-substituted cyclohexyl, oxetyl, optionally C 1-3 Alkyl or C 1-3 Halogenated alkyl-substituted piperidinyl, tetrahydropyranyl, or phenyl groups.

[0189] In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2A Each can be independently methyl, ethyl, or isopropyl, R 4G For optional use by R 4G1substituted with morpholinyl or -N(C 1-3 ethyl optionally substituted with methoxy or morpholinyl, n-propyl optionally substituted with C 1-3 alkyl, isobutyl optionally substituted with -OH or -OP(O)(OH)2, cyclopropyl, cyclobutyl, cyclopentyl, pyrrolidinyl optionally substituted with 1 to 3 groups each independently -NH2, C 1-6 alkyl or C 1-3 haloalkyl, oxetanyl, pyrrolidinyl optionally substituted with 1 to 3 methyl groups, piperidinyl optionally substituted with halogen or C 1-3 alkyl, tetrahydrofuranyl, tetrahydropyranyl or -C(O)C 1-6 alkyl, and R 4G1 is cyclopropyl, cyclobutyl, pyrrolidinyl optionally substituted with -NH2 or C 1-3 haloalkyl, oxetanyl, pyrrolidinyl optionally substituted with 1 to 3 methyl groups, piperidinyl optionally substituted with halogen or C 1-3 alkyl or C 1-3 haloalkyl, piperidinyl, tetrahydropyranyl or phenyl.

[0190] In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2A each independently is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl or isopentyl, and R 4G is methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl or 2-ethylbutyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2A each independently is methyl, ethyl or isopropyl, and R 4G is methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl or 2-ethylbutyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2AEach is a methyl group, and R 4G It is methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, or 2-ethylbutyl. In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2A Each is an ethyl group, and R 4G It is methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, or 2-ethylbutyl. In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2A Each is isopropyl, and R 4G It is methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, or 2-ethylbutyl. In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2A Each is independently methyl, ethyl, or isopropyl, and R 4G The compound is methyl. In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2A Each can be independently methyl, ethyl, or isopropyl, R 4G The compound is ethyl. In some embodiments, the compound may be represented by formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm) or (IIIn) or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2A Each can be independently methyl, ethyl, or isopropyl, R 4Gisopropyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2A each independently is methyl, ethyl, or isopropyl, R 4G is isopropyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2A each independently is methyl, ethyl, or isopropyl, R 4G is n-butyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2A each independently is methyl, ethyl, or isopropyl, R 4G is t-butyl. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2A each independently is methyl, ethyl, or isopropyl, R 4G is 2-ethylbutyl.

[0191] The compounds of the present disclosure include the compounds of Table 1A, Table 1B, Table 1C, Table 1D, Table 1E, Table 1F, Table 1G, Table 1H, and Table 1I. In some embodiments, the compound can be represented by Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the compounds in Table 1A, Table 1B, Table 1C, Table 1D, Table 1E, Table 1F, Table 1G, Table 1H, and Table 1I:

[0192] Table 1A. Compounds

[0193]

[0194]

[0195]

[0196]

[0197]

[0198]

[0199]

[0200] Table 1B. Compounds

[0201]

[0202]

[0203]

[0204]

[0205]

[0206]

[0207]

[0208]

[0209]

[0210] Table 1C. Compounds

[0211]

[0212] Table 1D. Compounds

[0213]

[0214]

[0215]

[0216]

[0217]

[0218]

[0219]

[0220]

[0221]

[0222] Table 1E. Compounds

[0223]

[0224]

[0225]

[0226]

[0227]

[0228]

[0229]

[0230]

[0231]

[0232] Table 1F. Compounds

[0233]

[0234]

[0235] Table 1G. Compounds

[0236]

[0237]

[0238]

[0239] Table 1H. Compounds

[0240]

[0241]

[0242] Table 1I. Compounds

[0243]

[0244]

[0245]

[0246]

[0247] In some embodiments, the compound of the disclosure or a pharmaceutically acceptable salt thereof has the following structure:

[0248]

[0249] In some embodiments, the compound of the disclosure or a pharmaceutically acceptable salt thereof has the following structure:

[0250]

[0251] In some embodiments, the compound of the disclosure or a pharmaceutically acceptable salt thereof, wherein the compound is:

[0252]

[0253]

[0254]

[0255] In some embodiments, the compound of the disclosure or a pharmaceutically acceptable salt thereof, wherein the compound is:

[0256]

[0257] In some embodiments, the compound of the disclosure or a pharmaceutically acceptable salt thereof, wherein the compound is:

[0258]

[0259] In some embodiments, the compound of the disclosure or a pharmaceutically acceptable salt thereof, wherein the compound is:

[0260]

[0261] In some embodiments, the compound of the disclosure or a pharmaceutically acceptable salt thereof, wherein the compound is:

[0262]

[0263] In some embodiments, the compound of the disclosure or a pharmaceutically acceptable salt thereof, wherein the compound is:

[0264]

[0265] In some embodiments, the compound of the disclosure or a pharmaceutically acceptable salt thereof, wherein the compound is:

[0266]

[0267] In some embodiments, the compound of the disclosure or a pharmaceutically acceptable salt thereof, wherein the compound is:

[0268]

[0269] In some embodiments, the compound of the disclosure or a pharmaceutically acceptable salt thereof, wherein the compound is:

[0270]

[0271] In vivo metabolites of the compounds described herein also fall within the scope of the present disclosure, to the extent such products are novel and unobvious in light of the prior art. Such products can result, for example, from the oxidation, reduction, hydrolysis, amidation, esterification, and the like of compounds administered to mammals, primarily due to enzymatic processes. Thus, the novel and unobvious compounds are produced by processes comprising contacting a compound with a mammal for a period of time sufficient to yield its metabolites. Such products typically are identified by preparing a radiolabeled (e.g., 14 C or 3 H) compound, administering a detectable dose (e.g., greater than about 0.5 mg / kg) of the radiolabeled compound to an animal, such as a rat, mouse, guinea pig, monkey, or man, allowing a sufficient time for metabolism to occur (typically about 30 seconds to 30 hours), and isolating the metabolites from urine, blood, or other biological samples. These metabolites are easily isolated since they are labeled, and (for other products) isolated by using antibodies capable of binding epitopes that survive in the metabolite. The metabolite structures are determined in conventional fashion, e.g., by MS or NMR analysis. Typically, the analysis of metabolites is performed in the same way as conventional drug metabolism studies. The conversion products, even if they themselves are not HSV antiviral active, are useful in diagnostic assays for therapeutic dosing of the compounds, as long as they are not otherwise found in vivo.

[0272] Formulations and methods for determining the stability of compounds in surrogate gastrointestinal secretions are known. Compounds are defined herein as stable in the gastrointestinal tract, wherein less than about 50 mole percent of protecting groups are deprotected in surrogate intestinal fluid or gastric fluid after incubation at 37°C for 1 hour. Just because a compound is stable to the gastrointestinal tract does not mean that they cannot be hydrolyzed in vivo. Prodrugs are generally stable in the digestive system, but can be substantially hydrolyzed to the parent drug in the digestive lumen, liver, lungs, or other metabolic organs, or generally within the cell. As used herein, a prodrug is understood to be a compound that is chemically designed to effectively release the parent drug after overcoming the biological barriers to oral delivery.

[0273] IV. PHARMACEUTICAL FORMULATIONS

[0274] In some embodiments, the present disclosure provides a pharmaceutical formulation comprising a pharmaceutically effective amount of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient. Also provided herein is a pharmaceutical formulation comprising a pharmaceutically effective amount of a compound of Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), and (IIn), or a pharmaceutically acceptable salt, solvate, and / or ester thereof, and a pharmaceutically acceptable carrier or excipient.

[0275] The compounds herein are formulated with conventional carriers and excipients, which will be selected according to routine procedures. Tablets will contain excipients, glidants, fillers, binders, and the like. Aqueous formulations are prepared in sterile form, and when intended for delivery by other than oral administration, generally will be isotonic. All formulations will optionally contain excipients such as those set forth in the "Handbook of Pharmaceutical Excipients" (1986). Excipients include ascorbic acid and other antioxidants, chelating agents such as EDTA, carbohydrates such as dextrin, hydroxyalkylcellulose, hydroxyalkylmethylcellulose, stearic acid and the like. The pH of the formulations will range from about 3 to about 11, but will generally be from about 7 to 10.

[0276] While it is possible for the active ingredient to be administered alone, it can be preferable to present it as a pharmaceutical formulation. Both veterinary and human formulations include at least one active ingredient as defined above, together with one or more acceptable carriers and optionally other therapeutic ingredients, especially those additional therapeutic ingredients discussed herein. The carrier(s) must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the subject in need thereof.

[0277] These formulations include those suitable for the aforementioned routes of administration. The formulations can conveniently be presented in unit dosage form and can be prepared by any of the methods well known in the art of pharmacy. Techniques and formulations generally are found in "Remington's Pharmaceutical Sciences" (Mack Publishing Co., Easton, PA). Such methods include the step of bringing into association the active ingredient with the carrier which constitutes one or more accessory ingredients. In general, the preparations are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product.

[0278] Formulations suitable for oral administration can be presented as discrete units such as capsules, sachets, or tablets each containing a pre-determined amount of the active ingredient; as a powder or granules; as a solution or a suspension in an aqueous or non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion. The active ingredient can also be administered as a bolus, electuary, or paste.

[0279] Tablets are made by compression or molding, optionally, with one or more accessory ingredients. Compressed tablets can be prepared by compressing, in a suitable machine, the active ingredient in a free-flowing form such as a powder or granules, optionally mixed with a binder, lubricant, inert diluent, preservative, surface-active or dispersing agent. Molded tablets can be made by molding, in a suitable machine, a mixture of the powdered active ingredient moistened with a liquid diluent. Tablets can optionally be coated or scored and optionally are formulated so as to provide slow or controlled release of the active ingredient therein.

[0280] For infections of the eye or other external tissues (e.g., mouth and skin), formulations are preferably applied as a topical ointment or cream containing the active ingredient(s) in an amount from 0.075 to 20% w / w (including active ingredients in the range of 0.1% and 20% between, in increments of 0.1% w / w, such as 0.6% w / w, 0.7% w / w, etc.), preferably 0.2% w / w to 15% w / w, and most preferably 0.5% w / w to 10% w / w. When formulated in an ointment, the active ingredient can be employed in a mixture with one or more of the following: a petrolatum base, a water miscible cream base, or a water miscible ointment base. Alternatively, the active ingredient can be formulated in a cream with an oil-in-water emulsion base or a water-in-oil emulsion base.

[0281] If desired, the aqueous phase of the cream base can comprise, for example, at least 30% w / w of a polyhydric alcohol, i.e. an alcohol having two or more hydroxyl groups such as propylene glycol, butane 1,3-diol, mannitol, sorbitol, glycerol and polyethylene glycol (including PEG 400) and mixtures thereof. Topical formulations can desirably include compounds which enhance the absorption or penetration of the active ingredients through the skin or other affected area. Examples of such skin penetration enhancers include dimethyl sulfoxide and related analogues.

[0282] The oil phase of the emulsion can be constructed in known manner from known ingredients. Whilst this phase can comprise only emulsifiers (or emulsifying agents), it desirably comprises a mixture of at least one emulsifier with a fat or oil or with both a fat and an oil. Preferably, a hydrophilic emulsifier is included together with a lipophilic emulsifier which acts as a stabiliser. It is also preferable to include both an oil and a fat. The emulsifiers with or without stabilisers together constitute so-called emulsifying waxes, and the waxes together with the oils and fats constitute so-called emulsifying soft paste bases which form the oily dispersed phase of the cream formulation.

[0283] Suitable emulsifiers and emulsion stabilisers for use in the formulation include Cetyl stearyl alcohol, benzyl alcohol, myristyl alcohol, glycerol monostearate and sodium lauryl sulphate.

[0284] Suitable oils or fats are chosen for the formulation based on the desired cosmetic properties to be achieved. The cream should preferably be a non-greasy, non-staining and washable product, with suitable consistency to avoid leakage from tubes or other containers. Straight or branched chain, mono- or di-alkyl esters of fatty acids can be used, such as diisohexyl adipate, isohexyl stearate, propylene glycol diester of coconut fatty acids, isopropyl myristate, decyl oleate, isopropyl palmitate, butyl stearate, palmitic acid-2-ethylhexyl ester or mixtures of branched esters known as Crodamol CAP, the last three being preferred esters. These esters can be used individually or in combination depending on the properties required. Alternatively, a high melting point lipid such as white soft paraffin and / or liquid paraffin or other mineral oil can be used.

[0285] The pharmaceutical formulations herein comprise a combination of an active ingredient with one or more pharmaceutically acceptable carriers or excipients and optionally other therapeutic agents. Pharmaceutical formulations containing the active ingredient can be in any form suitable for the intended method of administration. For example, when used for oral use the tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, solutions, syrups, or elixirs can be prepared. Compositions intended for oral use can be prepared according to any method known in the art for the manufacture of pharmaceutical compositions, and such compositions can contain one or more agents including sweetening agents, flavoring agents, coloring agents and preserving agents, in order to provide a palatable preparation. Tablets containing the active ingredient in admixture with non-toxic pharmaceutically acceptable excipient are also acceptable. These excipients can be, for example, inert diluents, such as calcium or sodium carbonate, lactose, calcium or sodium phosphate; granulating and disintegrating agents, such as corn starch, or alginic acid; binding agents, such as starch, gelatin or acacia; and lubricating agents such as magnesium stearate, stearic acid or talc. The tablets can be uncoated or they can be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over an extended period of time. For example, a time delay material such as glyceryl monostearate or glyceryl distearate can be employed.

[0286] Formulations for oral use can also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, or kaolin, or as soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, such as peanut oil, liquid paraffin or olive oil.

[0287] Aqueous suspensions contain the active material in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients include suspending agents, such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia; dispersing, or wetting agents such as a naturally occurring phosphatide (for example, lecithin), a condensation product of an alkylene oxide with a fatty acid (for example, polyoxyethylene stearate), a condensation product of an alkylene oxide with p-ester derived from fatty acids and hexitol anhydrides (for example, polyoxyethylene dehydrated sorbitol mono-oleate), and the like. The aqueous suspensions can also contain one or more preservatives, for example, ethyl or n-propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents and one or more sweetening agents, such as sucrose or saccharin.

[0288] Oil suspensions can be formulated by suspending the active ingredients in a vegetable oil (such as arachis oil, olive oil, sesame oil or coconut oil), or in a mineral oil (such as liquid paraffin). The oral suspensions can contain, together with the active ingredient, sweetening agents such as those described above, and flavouring agents. They can be preserved by the addition of an antioxidant (such as ascorbic acid) and can contain one or more components to protect the active ingredient against light, such as quinine, and to prevent the proliferation of microorganisms, such as benzalkonium chloride. The compositions can be in unit dosage forms, such as capsules, tablets, and suspensions, or can be presented as a dry powder for constitution with an appropriate vehicle, e.g. water, before use.

[0289] Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water are also possible. These compositions contain the active ingredient mixed with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those disclosed above. Additional excipients, for example sweetening, flavouring and colouring agents, can also be present.

[0290] The pharmaceutical compositions can also be in the form of oil-in-water emulsions. The oily phase can be a vegetable oil (such as olive oil or arachis oil), a mineral oil (such as liquid paraffin) or a mixture of these. Suitable emulsifying agents include naturally occurring gums, such as gum acacia and gum tragacanth, naturally occurring phosphatides, such as soybean lecithin, esters or partial esters derived from fatty acids and hexitol anhydrides, for example, sorbitan monooleate, and condensation products of these partial esters with ethylene oxide, for example, polyoxyethylene sorbitan monooleate. The emulsions can also contain sweetening and flavouring agents. Syrups and elixirs can be formulated with sweetening agents, for example glycerol, sorbitol or sucrose. Such formulations can also contain a demulcent, a preservative, flavouring or colouring agents.

[0291] The pharmaceutical compositions can be in the form of a sterile injectable aqueous or oleagenous suspension. This suspension can be formulated according to known art using those suitable dispersing or wetting agents and suspending agents which have been mentioned above. The sterile injectable preparation can also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol alone or in combination with a

[0292] The amount of active ingredient that can be combined with the carrier materials to produce a single dosage form will vary depending upon the host to be treated and the particular mode of administration. For example, a time-release formulation intended for oral administration to a human can contain approximately 1 mg to 1000 mg of active material compounded with an appropriate and convenient amount of carrier material to make a dosage unit form that is suitably adapted for the oral administration of the active ingredient to a human being. The active ingredient can be compounded for convenient and

[0293] Formulations suitable for topical application to the eye also include drops, in which the active ingredient is dissolved or suspended in a suitable carrier, especially an aqueous solvent for the active ingredient. The active ingredient preferably is present in such formulations in a concentration of 0.5 to 20%, advantageously 0.5 to 10%, and particularly about 1.5% w / w.

[0294] Formulations suitable for topical application to the eye also include drops, in which the active ingredient is dissolved or suspended in a suitable carrier, especially an aqueous solvent for the active ingredient. The active ingredient preferably is present in such formulations in a concentration of 0.5 to 20%, advantageously 0.5 to 10%, and particularly about 1.5% w / w.

[0295] Formulations for rectal administration can be provided as suppositories having a weight of about 1 to 2 g, with formulations containing about 0.5 to 500 mg of the active ingredient.

[0296] Formulations suitable for intrapulmonary or intranasal administration have a particle size that is selected from a group of particles having sizes in a range from 0.1 to 500 microns, such as 0.5, 1, 30, 35, and 50 microns, and are administered by rapid inhalation through the nasal passage or by inhalation into the oral cavity to reach the alveolar sacs. Suitable formulations include aqueous or oil solutions of the active ingredient. Formulations suitable for aerosol or dry powder administration can be prepared according to conventional methods and can deliver the active ingredient directly to the lungs, nose, or throat.

[0297] Another embodiment provides a novel, effective, safe, non-irritating, and physiologically compatible inhalable composition comprising a compound of formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) or a pharmaceutically acceptable salt thereof, the inhalable composition being suitable for treating pulmonary viral infections and potentially related bronchiolitis. Preferred pharmaceutically acceptable salts are inorganic acid salts, including hydrochlorides, hydrobromides, sulfates, or phosphates, as they cause less lung irritation. Preferably, the inhalable formulation is delivered into the bronchial space as an aerosol comprising particles with a median mass aerodynamic diameter (MMAD) between about 1 μm and about 5 μm. Preferably, compounds of formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIIn) are formulated for aerosol delivery using a nebulizer, a metered-dose inhaler (pMDI), or a dry powder inhaler (DPI).

[0298] Non-limiting examples of nebulizers include atomizing, jetting, ultrasonic, pressurizing, vibrating perforated plate, or equivalent nebulizers, including those utilizing adaptive aerosol delivery technologies (Denyer, J. Aerosol medicine Pulmonary Drug Delivery 2010, 23 Supplement 1, S1-S10). Jetting nebulizers use air pressure to break up a liquid solution into aerosol droplets. Ultrasonic nebulizers function by using piezoelectric crystals that shear the liquid into small aerosol droplets. Pressurized nebulizer systems force a solution under pressure through a small orifice to generate aerosol droplets. Vibrating perforated plate devices utilize rapid vibration to shear a liquid stream into appropriate droplet sizes.

[0299] In a preferred embodiment, the formulation of the compound of formula (II), (Ila), (lib), (lie), (lid), (lie), (Ilf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn) is nebulized to particles of the desired MMAD using a nebulizer capable of nebulizing the formulation of the compound of formula (II), (Ila), (lib), (lie), (lid), (lie), (Ilf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn) into particles of the desired MMAD, the formulation for nebulization is delivered to the intrabronchial space as an aerosol comprising particles with a MMAD predominantly between about 1 μm and about 5 μm. For optimal therapeutic effectiveness and to avoid upper airway and systemic side effects, the MMAD of the majority of the nebulized particles should not be greater than about 5 μm. If the aerosol contains a significant amount of particles with a MMAD greater than 5 μm, the particles will deposit in the upper airways, thereby reducing the amount of drug delivered to the site of inflammation and bronchoconstriction in the lower airways. If the MMAD of the aerosol is less than about 1 μm, the particles tend to remain suspended in the inhaled air and are subsequently exhaled during exhalation.

[0300] When formulated and delivered according to the methods herein, the aerosol formulation for nebulization delivers a therapeutically effective dose of the compound of formula (II), (Ila), (lib), (lie), (lid), (lie), (Ilf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn) to the site of infection with the family Pneumoviridae. The amount of drug administered must be adjusted to reflect the efficiency of delivery of the therapeutically effective dose of the compound of formula (II), (Ila), (lib), (lie), (lid), (lie), (Ilf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn). In a preferred embodiment, the combination of an aqueous aerosol formulation with a nebulizer, jet, pressurized, vibrating multi-orifice plate, or ultrasonic nebulizer allows for delivery of about at least 20% to about 90%, typically about 70%, of the administered dose of the compound of formula (II), (Ila), (lib), (lie), (lid), (lie), (Ilf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn) into the airways, depending on the nebulizer. In a preferred embodiment, at least about 30% to about 50% of the active compound is delivered. More preferably, about 70% to about 90% of the active compound is delivered.

[0301] In another embodiment, the compound of Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, is delivered as an inhalable dry powder. The compound is administered intrabronchially as a dry powder formulation to effectively deliver the fine particles of the compound into the intrabronchial space using a dry powder or metered dose inhaler. For delivery by DPI, the compound of Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn) is processed by mill jet milling, spray drying, critical fluid processing, or precipitation from solution into particles with a MMAD primarily between about 1 pm and about 5 pm. Media milling, jet milling, and spray drying devices and methods capable of producing particle sizes with a MMAD between about 1 pm and about 5 pm are well known in the art. In one embodiment, an excipient is added to the compound of Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn) prior to processing into particles of the desired size. In another embodiment, an excipient is blended with the particles of the desired size to aid in the dispersion of the drug particles, for example, by using lactose as an excipient.

[0302] Particle size determinations are made using devices well known in the art. For example, a multistage Anderson cascade impactor or other suitable method such as those specifically set forth in the United States Pharmacopeia Chapter 601, which is a device for the characterization of aerosols in metered dose inhalers and dry powder inhalers.

[0303] In another preferred embodiment, the compound of Formula (II), (Ila), (lib), (lie), (lid), (lie), (Ilf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn) is delivered in dry powder form using a device such as a dry powder inhaler or other dry powder dispersion device. Non-limiting examples of dry powder inhalers and devices include those disclosed in US 5,458,135; US 5,740,794; US 5775320; US 5,785,049; US 3,906,950; US 4,013,075; US 4,069,819; US 4,995,385; US 5,522,385; US 4,668,218; US 4,667,668; US 4,805,811; and US 5,388,572. Dry powder inhalers come in two main designs. One design is a metered device, where a reservoir for the drug is placed within the device, and the patient adds a dose of the drug to the inhalation chamber. The second design is a factory metered device, where each individual dose is prepared in a separate container. Both systems depend on the drug being formulated into small particles with a MMAD of 1 pm to about 5 pm, and often involve co-formulation with larger excipient particles such as, but not limited to, lactose. The drug powder is placed in the inhalation chamber (either by device metering or by breaking open a factory metered dose), and the patient’s inspiratory flow accelerates the powder out of the device and into the mouth. The non-laminar nature of the powder’s path causes the excipient-drug aggregates to break apart, and the mass of the large excipient particles causes them to impact in the back of the throat, while the smaller drug particles are deeply deposited in the lungs. In a preferred embodiment, the compound of Formula (II), (Ila), (lib), (lie), (lid), (lie), (Ilf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn), or a pharmaceutically acceptable salt thereof, is delivered in dry powder form using any type of dry powder inhaler as described herein, wherein the MMAD of the dry powder, which does not contain any excipient, is predominantly in the range of 1 pm to about 5 pm.

[0304] In another embodiment, the compound of Formula (II), (Ila), (lib), (lie), (lid), (lie), (Ilf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn) is delivered in dry powder form using a metered dose inhaler. Non-limiting examples of metered dose inhalers and devices include those disclosed in US 5,261,538; US 5,544,647; US 5,622,163; US 4,955,371; US 3,565,070; US 3,361306; and US 6,116,234. In a preferred embodiment, the compound of Formula (II), (Ila), (lib), (lie), (lid), (lie), (Ilf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn) or a pharmaceutically acceptable salt thereof is delivered in dry powder form using a metered dose inhaler, wherein the MMAD of the dry powder, which does not contain any excipients, is predominantly in the range of about 1-5 μm.

[0305] Formulations suitable for vaginal administration can be presented as pessaries, tampons, creams, gels, pastes, foams or spray formulations containing in addition to the active ingredient such carriers as are known in the art to be appropriate.

[0306] Formulations suitable for parenteral administration include aqueous and nonaqueous sterile injection solutions which can contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which can include suspending agents and thickening agents.

[0307] The formulations are presented in unit- or multi-dose containers, for example, sealed ampoules and vials, and can be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example water for injections, immediately prior to use. Extemporaneous injection solutions and suspensions are prepared from sterile powders, granules and tablets of the kind previously described. Preferred unit dosage formulations are those containing a daily dose or unit daily sub-dose, as herein above described, in an amount suitable for the subject being treated.

[0308] It will be appreciated that, in addition to the ingredients particularly mentioned above, the formulations can include other agents conventional in the art having regard to the type of formulation in question, for example those suitable for oral administration can include flavouring agents.

[0309] Also provided are veterinary compositions comprising at least one active ingredient as defined above together with a veterinary carrier.

[0310] The veterinary carriers are materials which can be used to administer the compositions, and can be solid, liquid or gaseous materials which are otherwise inert or acceptable in the veterinary art and compatible with the active ingredient. These veterinary compositions can be administered orally, parenterally or by any other desired route.

[0311] The compounds herein are used to provide controlled release pharmaceutical formulations ("controlled release formulations") comprising one or more of the compounds as the active ingredient, wherein the release of the active ingredient is controlled and modulated to allow for less frequent dosing or to improve the pharmacokinetic or toxicity profile of a given active ingredient.

[0312] The effective dosage of the active ingredient depends at least on the nature of the condition being treated, the toxicity, whether the compound is used prophylactically (lower dosage) or to combat an active viral infection, the method of delivery and the pharmaceutical formulation, and will be determined by the clinician using routine dose escalation studies. The dosage can be expected to be from about 0.0001 to about 100 mg / kg body weight per day; typically, from about 0.01 to about 10 mg / kg body weight per day; more typically, from about 0.01 to about 5 mg / kg body weight per day; most typically, from about 0.05 to about 0.5 mg / kg body weight per day. For example, a candidate daily dosage for an adult human weighing about 70 kg would range from 1 mg to 1000 mg, preferably from 5 mg to 500 mg, and can be given in single or multiple doses.

[0313] V. ROUTES OF ADMINISTRATION

[0314] One or more of the compounds of formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm) or (IIn) (referred to herein as the active ingredient) are administered by any route appropriate for the condition being treated. Suitable routes include oral, rectal, nasal, pulmonary, topical (including buccal and sublingual), vaginal and parenteral (including subcutaneous, intramuscular, intravenous, intradermal, intrathecal and epidural) and the like. It will be appreciated that the preferred route can vary with, for example, the condition being treated. An advantage of the compounds herein is that they are orally bioavailable and can be administered orally.

[0315] The compounds of the present disclosure (also referred to herein as the active ingredient) can be administered by any route appropriate for the condition being treated. Suitable routes include oral, rectal, nasal, topical (including buccal and sublingual), transdermal, vaginal and parenteral (including subcutaneous, intramuscular, intravenous, intradermal, intrathecal and epidural) and the like. It will be appreciated that the preferred route can vary with, for example, the condition being treated. An advantage of certain compounds disclosed herein is that they are orally bioavailable and can be administered orally.

[0316] The compounds of the present disclosure can be administered to an individual for a desired period of time or duration according to an effective dosing regimen, such as at least about one month, at least about 2 months, at least about 3 months, at least about 6 months, or at least about 12 months or more. In one variation, the compounds are administered on a daily or intermittent schedule over the course of an individual’s life.

[0317] The dosage or frequency of administration of the compounds of the present disclosure can be adjusted during the course of treatment according to the judgment of the administering physician.

[0318] The compounds can be administered to an individual (e.g., a human) in an effective amount. In some embodiments, the compounds are administered once per day.

[0319] The compounds can be administered by any available route and method, such as by oral or parenteral (e.g., intravenous) administration. A therapeutically effective amount of the compounds can include from about 0.00001 mg / kg body weight / day to about 10 mg / kg body weight / day, such as from about 0.0001 mg / kg body weight / day to about 10 mg / kg body weight / day, or such as from about 0.001 mg / kg body weight / day to about 1 mg / kg body weight / day, or such as from about 0.01 mg / kg body weight / day to about 1 mg / kg body weight / day, or such as from about 0.05 mg / kg body weight / day to about 0.5 mg / kg body weight / day, or such as from about 0.3 mg / day to about 30 mg / day, or such as from about 30 mg / day to about 300 mg / day.

[0320] The compounds of the present disclosure can be combined with one or more additional therapeutic agents in any dose of the compounds of the present disclosure (e.g., 1 mg to 1000 mg of the compound). A therapeutically effective amount can include about 1 mg / dose to about 1000 mg / dose, such as about 50 mg / dose to about 500 mg / dose, or such as about 100 mg / dose to about 400 mg / dose, or such as about 150 mg / dose to about 350 mg / dose, or such as about 200 mg / dose to about 300 mg / dose. Other therapeutically effective amounts of the compounds of the present disclosure are about 100 mg / dose, about 125 mg / dose, about 150 mg / dose, about 175 mg / dose, about 200 mg / dose, about 225 mg / dose, about 250 mg / dose, about 275 mg / dose, about 300 mg / dose, about 325 mg / dose, about 350 mg / dose, about 375 mg / dose, about 400 mg / dose, about 425 mg / dose, about 450 mg / dose, about 475 mg / dose, or about 500 mg / dose. Other therapeutically effective amounts of the compounds of the present disclosure are about 100 mg / dose, or about 125 mg / dose, about 150 mg / dose, about 175 mg / dose, about 200 mg / dose, about 225 mg / dose, about 250 mg / dose, about 275 mg / dose, about 300 mg / dose, about 350 mg / dose, about 400 mg / dose, about 450 mg / dose, or about 500 mg / dose. Single doses can be administered every hour, every day, or every week. For example, single doses can be administered once every 1, 2, 3, 4, 6, 8, 12, 16 hours, or once every 24 hours. Single doses can also be administered once every 1, 2, 3, 4, 5, 6 days, or once every 7 days. Single doses can also be administered once every 1, 2, 3 weeks, or once every 4 weeks. In some embodiments, single doses can be administered once a week. Single doses can also be administered once a month.

[0321] Other therapeutically effective amounts of the compounds of the present disclosure are about 20 mg / dose, 25 mg / dose, 30 mg / dose, 35 mg / dose, 40 mg / dose, 45 mg / dose, 50 mg / dose, 55 mg / dose, 60 mg / dose, 65 mg / dose, 70 mg / dose, 75 mg / dose, 80 mg / dose, 85 mg / dose, 90 mg / dose, 95 mg / dose, or about 100 mg / dose.

[0322] The frequency of dosing of the compounds of the present disclosure will be determined by the needs of the individual patient and can be, for example, once per day or twice or more per day. Administration of the compound continues for as long as the viral infection is being treated. For example, the compound can be administered to a person infected with a virus for a period of 20 days to 180 days, or for example, 20 days to 90 days, or for example, 30 days to 60 days.

[0323] The administration can be intermittent, with the patient receiving a daily dose of a compound of the disclosure for a period of several days or more, followed by a period of several days or more in which the patient does not receive a daily dose of the compound. For example, the patient can receive a dose of the compound every other day or three times per week. By way of further example, the patient can receive a dose of the compound daily for a period of 1 to 14 days, followed by a period of 7 to 21 days in which the patient does not receive a dose of the compound, followed by a subsequent period (e.g., 1 to 14 days) in which the patient again receives a daily dose of the compound. The alternating periods of administration and non-administration of the compound can be repeated as clinically indicated to treat the patient.

[0324] In one embodiment, a pharmaceutical composition is provided comprising a compound of the disclosure, or a pharmaceutically acceptable salt thereof, in combination with one or more (e.g., one, two, three, four, one or two, one to three, or one to four) additional therapeutic agents and a pharmaceutically acceptable excipient.

[0325] In one embodiment, a kit is provided comprising a compound of the disclosure, or a pharmaceutically acceptable salt thereof, in combination with one or more (e.g., one, two, three, four, one or two, one to three, or one to four) additional therapeutic agents.

[0326] In some embodiments, a compound of the disclosure, or a pharmaceutically acceptable salt thereof, is combined with one, two, three, four or more additional therapeutic agents. In some embodiments, a compound of the disclosure, or a pharmaceutically acceptable salt thereof, is combined with two additional therapeutic agents. In other embodiments, a compound of the disclosure, or a pharmaceutically acceptable salt thereof, is combined with three additional therapeutic agents. In other embodiments, a compound of the disclosure, or a pharmaceutically acceptable salt thereof, is combined with four additional therapeutic agents. The one, two, three, four or more additional therapeutic agents can be different therapeutic agents selected from the same class of therapeutic agents, and / or they can be selected from different classes of therapeutic agents.

[0327] In some embodiments, when a compound of the disclosure is combined with one or more additional therapeutic agents as described herein, the components of the combination are administered in a simultaneous or sequential regimen. When administered sequentially, the combination can be administered in two or more administrations.

[0328] In some embodiments, a compound of the disclosure is combined with one or more additional therapeutic agents in a single dosage form for simultaneous administration to a patient, for example, for oral administration as a solid dosage form.

[0329] In some embodiments, a compound of the disclosure is co-administered with one or more additional therapeutic agents.

[0330] To prolong the effect of the compounds of the present disclosure, it is often desirable to slow the absorption of the compound from subcutaneous or intramuscular injection. This can be accomplished by the use of a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the compound then depends upon its rate of dissolution, which, in turn, can depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered compound form is accomplished by dissolving or suspending the compound in an oil vehicle. Injectable depot forms are made by forming microencapsule matrices of the compound in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of compound to polymer and the particular polymer used, the rate of compound release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the compound in liposomes or microemulsions which are compatible with body tissues.

[0331] VI. COMBINATION THERAPIES

[0332] The compounds and compositions of Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn) provided herein are also used in combination with other active therapeutic agents for the treatment of viral infections, such as a pneumoviridae, picornaviridae, flaviviridae, or filoviridae viral infection.

[0333] Combination therapies for treating pneumoviridae

[0334] The compounds and compositions provided herein are also used in combination with other active therapeutic agents. For the treatment of a pneumoviridae viral infection, preferably, the other active therapeutic agent has activity against a pneumoviridae viral infection, in particular, a respiratory syncytial virus infection and / or a metapneumovirus infection. Non-limiting examples of these other active therapeutic agents with activity against RSV are ribavirin, palivizumab, motavizumab, RSV-IGIV (RespiGam®), MEDI-557, A-60444 (also known as RSV604), MDT-637, BMS-433771, ALN-RSV0, ALX-0171, and mixtures thereof. Other non-limiting examples of other active therapeutic agents with activity against respiratory syncytial virus infection include respiratory syncytial virus protein F inhibitors such as AK-0529; RV-521, ALX-0171, JNJ-53718678, BTA-585, and presatovir; RNA polymerase inhibitors such as lumicitabine and ALS-8112; anti-RSV G protein antibodies such as anti-G protein mAbs; viral replication inhibitors such as nitazoxanide.

[0335] ​In some embodiments, the other active therapeutic agent can be a vaccine for treating or preventing RSV, including but not limited to MVA-BN RSV, RSV-F, MEDI-8897, JNJ-64400141, DPX-RSV, SynGEM, GSK-3389245A, GSK-300389-1A, RSV-MEDI deltaM2-2 vaccine, VRC-RSVRGP084-00VP, Ad35-RSV-FA2, Ad26-RSV-FA2, and RSV fusion glycoprotein subunit vaccine.

[0336] Non-limiting examples of other active therapeutic agents having activity against metapneumovirus infection include sialidase modulators such as DAS-181; RNA polymerase inhibitors such as ALS-8112; and antibodies for treating metapneumovirus infection such as EV-046113.

[0337] In some embodiments, the other active therapeutic agent can be a vaccine for treating or preventing metapneumovirus infection, including but not limited to mRNA-1653 and rHMPV-Pa vaccine.

[0338] Combination therapies for treating picornaviridae

[0339] The compounds and compositions provided herein are also used in combination with other active therapeutic agents. For the treatment of Picornaviridae viral infections, preferably the other active therapeutic agent has activity against Picornaviridae viral infection, in particular enterovirus infection. Non-limiting examples of these other active therapeutic agents are capsid binding inhibitors such as pleconaril, BTA-798 (vapendavir), and other compounds disclosed by Wu et al. (US 7,078,403) and Watson (US 7,166,604); fusion sialidase proteins such as DAS-181; capsid protein VP1 inhibitors such as VVX-003 and AZN-001; viral protease inhibitors such as CW-33; phosphatidylinositol 4 kinase beta inhibitors such as GSK-480 and GSK-533; anti-EV71 antibodies.

[0340] In some embodiments, the other active therapeutic agent can be a vaccine for treating or preventing Picornaviridae viral infection, including but not limited to EV71 vaccine, TAK-021, and EV-D68 adenoviral vector-based vaccine.

[0341] Combination therapies for respiratory infections

[0342] Many infections of the pulmonary and picornaviridae virus families are respiratory tract infections. Thus, additional active therapeutic agents for treating respiratory symptoms and sequelae of infection can be used in combination with the compounds provided herein. The additional therapeutic agents are preferably administered orally or by direct inhalation. For example, other preferred additional therapeutic agents for treating viral respiratory infections in combination with the compounds provided herein include, but are not limited to, bronchodilators and corticosteroids.

[0343] Glucocorticoids

[0344] Glucocorticoids were first introduced as a therapy for asthma in 1950 (Carryer, Journal of Allergy, 21, 282-287, 1950) and remain the most effective and consistent therapy for the disease, but their mechanism of action has not been fully understood (Morris, J. Allergy Clin. Immunol., 75(1 Pt) 1-13, 1985). Unfortunately, oral glucocorticoid therapy is associated with serious adverse side effects such as truncal obesity, hypertension, glaucoma, glucose intolerance, accelerated cataract formation, bone mineral loss, and psychological effects, all of which limit their use as long-term therapeutic agents (Goodman and Gilman, 10th Edition, 2001). A solution to the systemic side effects is to deliver the steroid drug directly to the site of inflammation. Inhaled corticosteroids (ICS) have been developed to mitigate the serious side effects of oral steroids. Non-limiting examples of corticosteroids that can be used in combination with the compounds provided herein are dexamethasone, dexamethasone sodium phosphate, fluorometholone, fluorometholone acetate, loteprednol etabonate, loteprednol etabonate ophthalmic solution, hydrocortisone, prednisolone, flurandrenolide, triamcinolone, triamcinolone acetonide, betamethasone, beclomethasone dipropionate, methylprednisolone, fluocinolone, fluocinolone acetonide, flunisolide, fluocinbutte-21 -butylate, flumethasone, flumethasone pivalate, budesonide, halobetasol propionate, mometasone furoate, fluticasone, AZD-7594, ciclesonide; or a pharmaceutically acceptable salt thereof.

[0345] Anti-inflammatory agents

[0346] Other anti-inflammatory agents that act through anti-inflammatory cascades can also be used as additional therapeutic agents in combination with the compounds provided herein for the treatment of viral respiratory infections. The use of "anti-inflammatory signal transduction modulators" (referred to herein as AISTMs) such as phosphodiesterase inhibitors (e.g., PDE-4, PDE-5, or PDE-7 specific), transcription factor inhibitors (e.g., block NFκB through IKK inhibition), or kinase inhibitors (e.g., block P38 MAP, JNK, PI3K, EGFR, or Syk) is a rational approach to cutting off inflammation because these small molecules target a limited number of common intracellular pathways that are key points for anti-inflammatory therapeutic intervention (see review by P.J. Barnes, 2006). These non-limiting additional therapeutic agents include: 5-(2,4-difluoro-phenoxy)-l-isobutyl-lH-indazole-6-carboxylic acid (2-dimethylamino-ethyl)-amide (P38 Map kinase inhibitor ARRY-797); 3-cyclopropylmethoxy-N-(3,5-dichloro-pyridin-4-yl)-4-difluoromethoxy-benzamide (PDE-4 inhibitor Roflumilast); 4-[2-(3-cyclopentyloxy-4-methoxyphenyl)-2-phenyl-ethyl]-pyridine (PDE-4 inhibitor CDP-840); N-(3,5-dichloro-4-pyridinyl)-4-difluoromethoxy-8-[(methylsulfonyl)amino]-l-dibenzofurancarboxamide (PDE-4 inhibitor Oglemilast); N-(3,5-dichloro-pyridin-4-yl)-2-[l-(4-fluorobenzyl)-5-hydroxy-lH-indol-3-yl]-2-oxo-acetamide (PDE-4 inhibitor AWD 12-281); 8-methoxy-2-trifluoromethyl-quinoline-5-carboxylic acid (3,5-dichloro-l-oxo-pyridin-4-yl)-amide (PDE-4 inhibitor Sch 351591); 4-[5-(4-fluorophenyl)-2-(4-methanesulfinyl-phenyl)-lH-imidazol-4-yl]-pyridine (P38 inhibitor SB-203850); 4-[4-(4-fluorophenyl)-l-(3-phenylpropyl)-5-pyridin-4-yl-lH-imidazol-2-yl]-but-3-yn-1-ol (P38 inhibitor RWJ-67657); 4-cyano-4-(3-cyclopentyloxy-4-methoxy-phenyl)-cyclohexanecarboxylic acid 2-diethylamino-ethyl ester (2-diethyl-ethyl ester prodrug of Cilomilast, PDE-4 inhibitor); (3-chloro-4-fluorophenyl)-[7-methoxy-6-(3-morpholin-4-yl-propoxy)-quinazolin-4-yl]-amine (Gefitinib, EGFR inhibitor); and 4-(4-methyl-piperazin-l-ylmethyl)-N-[4-methyl-3-(4-pyridin-3-yl-pyrimidin-2-ylamino)-phenyl]-benzamide (Imatinib, EGFR inhibitor).

[0347] Beta2-adrenergic receptor agonist bronchodilators

[0348] Combinations comprising inhaled beta2-adrenergic receptor agonist bronchodilators such as formoterol, salbutamol or salmeterol in combination with the compounds provided herein are also suitable, but not limiting, combinations useful for treating respiratory viral infections.

[0349] Combinations of inhaled beta2-adrenergic receptor agonist bronchodilators such as formoterol or salmeterol in combination with ICS are also used to treat bronchoconstriction and inflammation (respectively and ). Combinations comprising these ICS and beta2-adrenergic receptor agonist combinations in combination with the compounds provided herein are also suitable, but not limiting, combinations useful for treating respiratory viral infections.

[0350] Other examples of beta2 adrenergic receptor agonists are bevonol, vilanterol, indacaterol, olodaterol, tulobuterol, formoterol, abinterol, salbutamol, arformoterol, levosalbutamol, fenoterol and TD-5471.

[0351] Anticholinergic agents

[0352] Anticholinergic agents have potential use for the treatment or prevention of bronchoconstriction in the lungs and thus can be used in combination with the compounds provided herein as an additional therapeutic agent for the treatment of viral respiratory infections.These anticholinergic agents include, but are not limited to, antagonists of muscarinic receptors (particularly the M3 subtype) that have proven therapeutic efficacy in humans for the control of cholinergic tone in COPD (Witek, 1999); 1-{4-hydroxy-1-[3,3,3-tris-(4-fluorophenyl)-propionyl]-pyrrolidine-2-carbonyl}- pyrrolidine-2-carboxylic acid (1-methyl-piperidin-4-ylmethyl)-amide; 3-[3-(2-diethylamino- acetyloxy)-2-phenyl-propionyloxy]-8-isopropyl-8-methyl-8-azonio-bicyclo[3.2.1]octane (isopropylamine-N,N-diethylglycine); 1-cyclohexyl-3,4-dihydro-1H-isoquinoline-2-carboxylic acid 1-aza-bicyclo[2.2.2]oct-3-yl ester (solfenazin); 2-hydroxymethyl-4-methane-sulfinyl-2-phenyl-butyric acid 1-aza-bicyclo[2.2.2]oct-3-yl ester (revatropate); 2-{1-[2-(2,3-dihydro-benzofuran-5-yl)-ethyl]-pyrrolidin-3-yl}-2,2-diphenyl-acetamide (darifenacin); 4-azepan-1-yl-2,2-diphenyl-butyramide (buzepide); 7-[3-(2-diethylamino-acetyloxy)-2-phenyl-propionyloxy]-9-ethyl-9-methyl-3-oxa-9-azonio- tricyclo[3.3.1.02,4]nonane (oxitropium-N,N-diethylglycine); 7-[2-(2-diethylamino-acetyloxy)-2,2-di-thiophen-2-yl-acetyloxy]-9,9-dimethyl-3-oxa-9-azonio- tricyclo[3.3.1.02,4]nonane (tiotropium-N,N-diethylglycine); dimethylamino-acetic acid 2-(3-diisopropylamino-1-phenyl-propyl)-4-methyl-phenyl ester (tolterodine-N,N- dimethylglycine); 3-[4,4-bis-(4-fluorophenyl)-2-oxo-imidazolidin-1-yl]-1-methyl-1-(2-oxo-2-pyridin-2-yl-ethyl)- pyrrolidinium; 1-[1-(3-fluorobenzyl)-piperidin-4-yl]-4,4-bis-(4-fluorophenyl)-imidazolidin-2-one; 1-cyclooctyl-3-(3-methoxy-1-aza-bicyclo[2.2.2]oct-3-yl)-1-phenyl-prop-2-yn-1-ol; 3-[2-(2-diethylamino-acetyloxy)-2,2-dithiophen-2-yl-acetyloxy]-1-(3-phenoxy-propyl)-1-azonio- bicyclo[2.2.2]octane (atropine-N,N-diethylglycine); or (2-diethylamino-acetyloxy)-dithiophen-2-yl-acetic acid 1-methyl-1-(2-phenoxy-ethyl)-piperidin-4-yl ester; rafenamin, glycopyrronium, umclidinium, tiotropium, aclidinium, benzquinonium.

[0353] Mucolytic agents

[0354] The compounds provided herein and the compositions provided herein can also be combined with mucolytic agents to treat infections and symptoms of respiratory infections. A non-limiting example of a mucolytic agent is ambroxol. Similarly, the compounds of Formula (I), (la), (lb), (Ic), (Id), (Ie), (If), (Ig), (Ih), (Ij), (Ik), (Im), or (In) can be combined with expectorants to treat infections and symptoms of respiratory infections. A non-limiting example of an expectorant is guaifenesin.

[0355] Nebulized hypertonic saline is used to improve immediate and long-term clearance of small airways in patients with lung disease (Kuzik, Pediatrics 2007, 266). Thus, the compounds provided herein can also be combined with nebulized hypertonic saline, particularly when a pneumoviridae virus infection is complicated by bronchiolitis. The combination of a compound of Formula (I) or Formula (II) with hypertonic saline can also include any of the additional agents described above. In one embodiment, about 3% hypertonic saline is used, which is nebulized.

[0356] Combination therapies for treating COPD

[0357] The compounds and compositions provided herein are also used in combination with other active therapeutic agents. For the treatment of respiratory exacerbations of COPD, other active therapeutic agents include other active agents against COPD. Non-limiting examples of these other active therapeutic agents include anti-IL5 antibodies such as benralizumab, mepolizumab; dipeptidyl peptidase I (DPP1) inhibitors such as AZD-7986 (INS-1007); DNA gyrase inhibitors / topoisomerase IV inhibitors such as ciprofloxacin hydrochloride; MDR- associated protein 4 / phosphodiesterase (PDE) 3 and 4 inhibitors such as RPL-554; CFTR stimulators such as inavolisib, QBW-251; MMP-9 / MMP-12 inhibitors such as RBx-10017609; adenosine A1 receptor antagonists such as PBF-680; GATA 3 transcription factor inhibitors such as SB-010; muscarinic receptor modulators / nicotine acetylcholine receptor agonists such as ASM-024; MARCKS protein inhibitors such as BIO-11006; kit tyrosine kinases / PDGF inhibitors such as masitinib; phosphodiesterase (PDE) 4 inhibitors such as roflumilast, CHF-6001; phosphoinositide-3 kinase delta inhibitors such as nemiralisib; 5-lipoxygenase inhibitors such as TA-270; muscarinic receptor antagonists / beta2 adrenergic receptor agonists such as bevonterol succinate, AZD-887, ipratropium bromide; TRN-157; elastase inhibitors such as eprotastat sodium; metalloproteinase-12 inhibitors such as FP-025; interleukin 18 ligand inhibitors such as tadekinig alfa; skeletal muscle troponin activators such as CK-2127107; p38 MAP kinase inhibitors such as acumapimod; IL-17 receptor modulators such as CNTO-6785; CXCR2 chemokine antagonists such as danirixin; leukocyte elastase inhibitors such as POL-6014; epoxide hydrolase inhibitors such as GSK-2256294; HNE inhibitors such as CHF-6333; VIP agonists such as apadenox; phosphoinositide-3 kinase delta / gamma inhibitors such as RV-1729; complement C3 inhibitors such as APL-1; and G protein-coupled receptor-44 antagonists such as AM-211.

[0358] Other non-limiting examples of active therapeutic agents also include budesonide, adipocell, nitric oxide, PUR-1800, YLP-001, LT-4001, azithromycin, gamunex, QBKPN, sodium pyruvate, MUL-1867, mannitol, MV-130, MEDI-3506, BI-443651, VR-096, OPK-0018, TEV-48107, doxofylline, TEV-46017, OligoG-COPD-5 / 20, ZP-051, lysine acetylsalicylate.

[0359] In some embodiments, the other active therapeutic agent can be a vaccine with anti-COPD activity, including but not limited to MV-130 and GSK-2838497A.

[0360] Combination therapies for treating Dengue fever

[0361] The compounds and compositions provided herein are also used in combination with other active therapeutic agents. For the treatment of Flaviviridae viral infections, preferably the other active therapeutic agent has activity against Flaviviridae viral infections, in particular dengue infections. Non-limiting examples of these other active therapeutic agents are host cytokine modulators such as GBV-006; fenretinide ABX-220, BRM-211; alpha-glucosidase 1 inhibitors such as sisguvacine; platelet-activating factor receptor (PAFR) antagonists such as modipafant; cadherin-5 / factor la modulators such as FX-06; NS4B inhibitors such as JNJ-8359; viral RNA splicing modulators such as ABX-202; NS5 polymerase inhibitors; NS3 protease inhibitors; and TLR modulators.

[0362] In some embodiments, the other active therapeutic agent can be a vaccine for the treatment or prevention of dengue, including but not limited to TetraVax-DV, DPIV-001, TAK-003, live attenuated dengue vaccine, tetravalent dengue vaccine, tetravalent DNA vaccine, rDEN2delta30-7169; and DENV-1 PIV.

[0363] Combination therapies for treating Ebola

[0364] The compounds and compositions provided herein are also used in combination with other active therapeutic agents. For the treatment of Filoviridae viral infections, preferably the other active therapeutic agent has activity against Filoviridae viral infections, in particular Marburg virus, Ebola virus, and Kyssa virus infections. Non-limiting examples of these other active therapeutic agents are: ribavirin, palivizumab, motavizumab, RSV-IGIV MEDI-557, A-60444, MDT-637, BMS-433771, amiodarone, dronedarone, verapamil, Ebola convalescent plasma (ECP), TKM-100201, BCX4430 ((2S,3S,4R,5R)-2-(4-amino-5H-pyrrolo[3,2-d]pyrimidin-7-yl)-5- (hydroxymethyl)pyrrolidine-3,4-diol), TKM-Ebola, T-705 monophosphate, T-705 diphosphate, T-705 triphosphate, FGI-106 (1-N,7-N-bis[3-(dimethylamino)propyl]-3,9- dimethylquinolo[8,7-h]quinolin-1,7-diamine), rNAPc2, OS-2966, brincidofovir, remdesivir; RNA polymerase inhibitors such as Galidesivir, favipiravir (also known as T-705 or Avigan), JK-05; host cytokine modulators such as GMV-006; Cadherin-5 / Factor la modulators such as FX-06; and antibodies for treating Ebola such as REGN-3470-3471-3479 and ZMapp.

[0365] Other non-limiting active therapeutic agents with anti-Ebola activity include alpha-glucosidase 1 inhibitors, cathepsin B inhibitors, CD29 antagonists, dendritic ICAM-3 grabbing nonintegrin 1 inhibitors, estrogen receptor antagonists, factor VII antagonists, HLA class II antigen modulators, host cytokine modulators, interferon alpha ligands, neutral alpha glucosidase AB inhibitors, Niemann-Pick Cl protein inhibitors, nucleoprotein inhibitors, polymerase cofactor VP35 inhibitors, serine protease inhibitors, tissue factor inhibitors, TLR-3 agonists, viral envelope glycoprotein inhibitors, and Ebola virus entry inhibitors (NPC1 inhibitors).

[0366] In some embodiments, other active therapeutic agents can be vaccines for treating or preventing Ebola, including but not limited to VRC-EBOADC076-00-VP, an adenovirus-based Ebola vaccine, rVSV-EBOV, rVSVN4CT1-EBOVGP, MVA-BN Filo + Ad26-ZEBOV regimen, INO-4212, VRC-EBODNA023-00-VP, VRC-EBOADC069-00-VP, GamEvac-combi vaccine, SRC VB vector, HPIV3 / EboGP vaccine, MVA-EBOZ, Ebola recombinant glycoprotein vaccine, Vaxart adenovirus vector 5-based Ebola vaccine, FiloVax vaccine, GOVX-E301, and GOVX-E302.

[0367] The compounds and compositions provided herein can also be used in combination with phosphoramidite morpholino oligomers (PMOs), which are synthetic antisense oligonucleotide analogs designed to interfere with the translation process by forming base-paired duplexes with specific RNA sequences. Examples of PMOs include, but are not limited to, AVI-7287, AVI-7288, AVI-7537, AVI-7539, AVI-6002, and AVI-6003.

[0368] The compounds and compositions provided herein are also intended for use in the general care of patients with a Filoviridae virus infection, including parenteral fluids (including dextrose saline and lactated Ringer's) and nutrients, antibiotics (including metronidazole and cephalosporin antibiotics such as ceftriaxone and cefuroxine) and / or antifungal prophylactics, fever and pain medications, antiemetics (such as metoclopramide) and / or antidiarrheal medications, vitamin and mineral supplements (including vitamin K and zinc sulfate), anti-inflammatory agents (such as ibuprofen), pain medications, and medications for other common ailments in the patient population, such as anti-malarial agents (including artemether and artemether-lumefantrine combination therapy), typhoid fever vaccines (including quinolone antibiotics such as ciprofloxacin, macrolide antibiotics such as azithromycin, cephalosporin antibiotics such as ceftriaxone, or aminopenicillins such as ampicillin), or Shigella disease vaccines.

[0369] VII. METHODS OF TREATING VIRAL INFECTIONS

[0370] The present disclosure provides methods of using compounds of Formula (I), (Ia), (Ib), (Ic), (Id), (Ie), (If), (Ig), (Ih), (Ij), (Ik), (Im), or (In) to treat a variety of diseases such as respiratory syncytial virus (RSV), Ebola virus, Zika virus, West Nile virus, Dengue fever, and HCV.

[0371] Paramyxoviridae

[0372] In some embodiments, the present disclosure provides methods for treating Paramyxoviridae infection, comprising administering to an individual (e.g., a human) infected with a Paramyxoviridae virus a therapeutically effective amount of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof. Paramyxoviridae viruses include, but are not limited to, Nipah virus and parainfluenza virus.

[0373] Pneumoviridae

[0374] In some embodiments, the present disclosure provides a method of treating a pneumoviridae virus infection in a human in need thereof, comprising administering to the human a therapeutically effective amount of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof. Pneumoviridae viruses include, but are not limited to, respiratory syncytial virus and human metapneumovirus. In some embodiments, the pneumoviridae virus infection is a respiratory syncytial virus infection. In some embodiments, the pneumoviridae virus infection is a human metapneumovirus infection.

[0375] In some embodiments, the present disclosure provides a method of making a medicament for treating a pneumoviridae virus infection in a human in need thereof, characterized by the use of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof. In some embodiments, the present disclosure provides the use of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for treating a pneumoviridae virus infection in a human. In some embodiments, the pneumoviridae virus infection is a respiratory syncytial virus infection. In some embodiments, the pneumoviridae virus infection is a human metapneumovirus infection.

[0376] In some embodiments, the present disclosure provides a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, for use in treating a pneumoviridae virus infection in a human in need thereof. In some embodiments, the pneumoviridae virus infection is a respiratory syncytial virus infection. In some embodiments, the pneumoviridae virus infection is a human metapneumovirus infection.

[0377] In some embodiments, the present disclosure provides a method for treating an RSV infection, comprising administering to an individual (e.g., a human) infected with respiratory syncytial virus a therapeutically effective amount of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof. Typically, the individual has a chronic respiratory syncytial virus infection, but treating an acutely infected human with RSV is also within the scope of the present disclosure.

[0378] In some embodiments, a method of inhibiting RSV replication is provided, comprising administering to an individual (e.g., a human) a compound of the present disclosure, or a pharmaceutically acceptable salt thereof.

[0379] In some embodiments, the present disclosure provides a method for reducing viral load associated with an RSV infection, wherein the method comprises administering to an individual (e.g., a human) infected with RSV a therapeutically effective amount of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, wherein the therapeutically effective amount is sufficient to reduce the RSV viral load in the individual.

[0380] As described more fully herein, a compound of the present disclosure can be administered to an individual (e.g., a human) infected with RSV along with one or more additional therapeutic agents. The additional therapeutic agent(s) can be administered to the infected individual (e.g., a human) simultaneously with, or prior to, or subsequent to, the compound of the present disclosure.

[0381] In some embodiments, provided are compounds of the disclosure, or pharmaceutically acceptable salts thereof, for use in treating or preventing an RSV infection. In some embodiments, provided are compounds of the disclosure (e.g., a compound of Formula (I), (la), (lb), (lc), (Id), (le), (If), (lg), (lh), (lj), (Ik), (Im), or (In)), or a pharmaceutically acceptable salt thereof, for use in the manufacture of a medicament for treating or preventing an RSV infection.

[0382] As described more fully herein, compounds of the disclosure can be administered to an individual (e.g., a human) infected with RSV with one or more additional therapeutic agents. Additionally, in some embodiments, when used to treat or prevent RSV, a compound of the disclosure can be administered with one or more (e.g., one, two, three, four, or more) additional therapeutic agents selected from the group consisting of RSV combination drugs, RSV vaccines, RSV DNA polymerase inhibitors, immunomodulators toll-like receptor (TLR) modulators, interferon alpha receptor ligands, hyaluronidase inhibitors, respiratory syncytial surface antigen inhibitors, cytotoxic T-lymphocyte-associated protein 4 (ipil4) inhibitors, cyclophilin inhibitors, RSV viral entry inhibitors, antisense oligonucleotides targeting viral mRNA, short interfering RNA (siRNA) and ddRNAi endonuclease modulators, ribonucleotide reductase inhibitors, RSV E antigen inhibitors, covalently closed circular DNA (cccDNA) inhibitors, farnesol X receptor agonists, RSV antibodies, CCR2 chemokine antagonists, thymosin agonists, cytokines, nucleoprotein modulators, retinoic acid-inducible gene 1 stimulators, NOD2 stimulators, phosphoinositide 3-kinase (PI3K) inhibitors, indoleamine-2,3-dioxygenase (IDO) pathway inhibitors, PD-1 inhibitors, PD-L1 inhibitors, recombinant thymosin alpha-1, Bruton’s tyrosine kinase (BTK) inhibitors, KDM inhibitors, RSV replication inhibitors, arginase inhibitors, and other RSV drugs.

[0383] Picornaviridae

[0384] In some embodiments, the present disclosure provides a method of treating a Picornaviridae virus infection in a human in need thereof, comprising administering to said human a therapeutically effective amount of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof. Picornaviridae viruses are enteroviruses that cause heterogeneous infections including herpangina, aseptic meningitis, common cold-like syndrome (human rhinovirus infection), nonparalytic poliomyelitis-like syndrome, epidemic myalgia (an acute, febrile, infectious illness that occurs in epidemics), hand, foot, and mouth syndrome, pediatric and adult pancreatitis, and severe myocarditis. In some embodiments, the Picornaviridae virus infection is a human rhinovirus infection.

[0385] In some embodiments, the present disclosure provides a method of making a medicament for treating a Picornaviridae virus infection in a human in need thereof, characterized by the use of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof. In some embodiments, the present disclosure provides the use of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for treating a Picornaviridae virus infection in a human. In some embodiments, the Picornaviridae virus infection is a human rhinovirus infection.

[0386] In some embodiments, the present disclosure provides a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, for use in treating a Picornaviridae virus infection in a human in need thereof. In some embodiments, the Picornaviridae virus infection is a human rhinovirus infection.

[0387] Flaviviridae

[0388] In some embodiments, the present disclosure provides a method of treating a Flaviviridae virus infection in a human in need thereof, comprising administering to said human a therapeutically effective amount of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof. Representative Flaviviridae viruses include, but are not limited to, dengue fever, yellow fever, West Nile virus, Zika virus, Japanese encephalitis virus, and hepatitis C virus (HCV). In some embodiments, the Flaviviridae virus infection is a dengue fever virus infection. In some embodiments, the Flaviviridae virus infection is a yellow fever virus infection. In some embodiments, the Flaviviridae virus infection is a West Nile virus infection. In some embodiments, the Flaviviridae virus infection is a Zika virus infection. In some embodiments, the Flaviviridae virus infection is a Japanese encephalitis virus infection. In some embodiments, the Flaviviridae virus infection is a hepatitis C virus infection.

[0389] In some embodiments, the present disclosure provides a method of preparing a medicament for treating a Flaviviridae virus infection in a human in need thereof, characterized by use of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof. In some embodiments, the present disclosure provides use of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, for the preparation of a medicament for treating a Flaviviridae virus infection in a human. In some embodiments, the Flaviviridae virus infection is a dengue virus infection. In some embodiments, the Flaviviridae virus infection is a yellow fever virus infection. In some embodiments, the Flaviviridae virus infection is a West Nile virus infection. In some embodiments, the Flaviviridae virus infection is a Zika virus infection. In some embodiments, the Flaviviridae virus infection is a hepatitis C virus infection.

[0390] In some embodiments, the present disclosure provides a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, for use in treating a Flaviviridae virus infection in a human in need thereof. In some embodiments, the Flaviviridae virus infection is a dengue virus infection. In some embodiments, the Flaviviridae virus infection is a yellow fever virus infection. In some embodiments, the Flaviviridae virus infection is a West Nile virus infection. In some embodiments, the Flaviviridae virus infection is a Zika virus infection. In some embodiments, the Flaviviridae virus infection is a hepatitis C virus infection.

[0391] Filoviridae

[0392] In some embodiments, the present disclosure provides a method of treating an Filoviridae virus infection in a human in need thereof, comprising administering to said human a therapeutically effective amount of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof. Representative Filoviridae viruses include, but are not limited to, Ebola virus and Marburg virus. In some embodiments, the Filoviridae virus infection is an Ebola virus infection.

[0393] In some embodiments, the present disclosure provides a method of preparing a medicament for treating a Filoviridae virus infection in a human in need thereof, characterized by use of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof. In some embodiments, the present disclosure provides use of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, for the preparation of a medicament for treating a Filoviridae virus infection in a human. In some embodiments, the Filoviridae virus infection is an Ebola virus infection.

[0394] In some embodiments, the present disclosure provides a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, for use in treating a Filoviridae virus infection in a human in need thereof. In some embodiments, the Filoviridae virus infection is an Ebola virus infection.

[0395] VIII. METHODS OF TREATING OR PREVENTING WORSENING OF RESPIRATORY CONDITIONS CAUSED BY VIRAL INFECTIONS

[0396] The compound of Formula (II), (IIa), (IIb), (IIc), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (IIj), (IIk), (IIm), or (IIn) can also be used to treat or prevent exacerbation of a respiratory condition caused by a viral infection in a human in need thereof.

[0397] In some embodiments, the present disclosure provides a method for treating or preventing exacerbation of a respiratory condition caused by a viral infection in a human in need thereof, comprising administering to the human a therapeutically effective amount of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, wherein the respiratory condition is chronic obstructive pulmonary disease. In some embodiments, the viral infection is caused by respiratory syncytial virus, rhinovirus, or metapneumovirus.

[0398] In some embodiments, the present disclosure provides a method for treating or preventing exacerbation of a respiratory condition caused by a viral infection in a human in need thereof, comprising administering to the human a therapeutically effective amount of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, wherein the respiratory condition is asthma. In some embodiments, the viral infection is caused by respiratory syncytial virus, rhinovirus, enterovirus, or metapneumovirus.

[0399] In some embodiments, the present disclosure provides a method of manufacturing a medicament for treating or preventing exacerbation of a respiratory condition caused by a viral infection in a human in need thereof, characterized in that a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, is used, wherein the respiratory condition is chronic obstructive pulmonary disease. In some embodiments, the viral infection is caused by respiratory syncytial virus, rhinovirus, or metapneumovirus.

[0400] In some embodiments, the present disclosure provides a method of manufacturing a medicament for treating or preventing exacerbation of a respiratory condition caused by a viral infection in a human in need thereof, characterized in that a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, is used, wherein the respiratory condition is asthma. In some embodiments, the viral infection is caused by respiratory syncytial virus, rhinovirus, enterovirus, or metapneumovirus.

[0401] In some embodiments, the present disclosure provides use of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for treating or preventing exacerbation of a respiratory condition caused by a viral infection in a human, wherein the respiratory condition is chronic obstructive pulmonary disease. In some embodiments, the viral infection is caused by respiratory syncytial virus, rhinovirus, or metapneumovirus.

[0402] In some embodiments, the present disclosure provides use of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment or prevention of exacerbation of a respiratory condition caused by a viral infection in a human, wherein the respiratory condition is asthma. In some embodiments, the viral infection is caused by respiratory syncytial virus, rhinovirus, enterovirus, or metapneumovirus.

[0403] In some embodiments, the present disclosure provides a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, for use in the treatment or prevention of exacerbation of a respiratory condition caused by a viral infection in a human in need thereof, wherein the respiratory condition is chronic obstructive pulmonary disease. In some embodiments, the viral infection is caused by respiratory syncytial virus, rhinovirus, or metapneumovirus.

[0404] In some embodiments, the present disclosure provides a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, for use in the treatment or prevention of exacerbation of a respiratory condition caused by a viral infection in a human in need thereof, wherein the respiratory condition is asthma. In some embodiments, the viral infection is caused by respiratory syncytial virus, rhinovirus, enterovirus, or metapneumovirus.

[0405] IX. EXAMPLES

[0406] Abbreviations. Certain abbreviations and acronyms are used throughout the experimental details. Although most of these abbreviations and acronyms are understood by those skilled in the art, Table 2 contains a list of many of these abbreviations and acronyms.

[0407] Table 2. List of Abbreviations and Acronyms .

[0408]

[0409]

[0410] Compounds were subjected to preparative HPLC (Phenomenex Gemini 10u C18 AXIA 250 x 21.2 mm column with 30-70% acetonitrile / water with 0.1% TFA gradient). Some compounds were obtained as TFA salts following this preparative HPLC method.

[0411] Compounds using the "P a " or "P b " designation refer to the (R)- or (S)-isomer, where the specific stereochemistry at this position is not specified.

[0412] A. INTERMEDIATES

[0413] Intermediate 1. (2R,3S,4R,5S)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-3,4- dihydroxy-2-(hydroxymethyl)tetrahydrofuran-2-carbonitrile Intermediate 2. tert-Butyl (7-((3aS,4S,6R,6aS)-6-cyano-6-(hydroxymethyl)-2,2- dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)pyrrolo[2,1-f][1,2,4]triazin-4-yl)carbamate

[0414]

[0415] This product can be prepared according to WO 2015 / 069939. For example, pages 43-54 of WO 2015 / 069939 provide a method for preparing the compound identified as Compound 1 in WO 2015 / 069939.

[0416] Intermediate 3. (3aS,4R,6S,6aS)-6-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-4- (((tert-butyldimethylsilyl)oxy)methyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxole-4- carbonitrile Intermediate 4. (3aS,4R,6S,6aS)-6-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-4- (hydroxymethyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxole-4-carbonitrile

[0417]

[0418] Compound 14j from WO 2015 / 069939 (21.79 g, 39.93 mmol) in THF (400 mL) was cooled in an ice bath. TBAF 1.0 M in THF (50.0 mL, 50.0 mmol) was added in one portion. The mixture was allowed to reach ambient temperature and stirred for about 30 min. The reaction was determined to be complete by LCMS. The reaction mixture was quenched with water and the organics were removed under reduced pressure. The crude product was partitioned between EtOAc and water. The layers were separated and the aqueous phase was washed with EtOAc. The organics were combined and dried over sodium sulfate. The solids were filtered off and the solvent was removed under reduced pressure. The crude product was purified by silica gel chromatography (330 g column, 30% - 100% EtOAc in hexanes) to give the product. MS m / z = 431.74 [M+1]. 1 H NMR (400 MHz, DMSO-d6) δ 10.53 (s, 1H), 8.25 (s, 1H), 7.21 (s, 1H), 7.03 (d, J = 4.6 Hz, 1H), 5.77 (t, J = 6.1 Hz, 1H), 5.59 (d, J = 4.0 Hz, 1H), 5.27 (dd, J = 6.7, 4.1 Hz, 1H), 4.94 (d, J = 6.7 Hz, 1H), 3.66 (dd, J = 6.1, 2.4 Hz, 2H), 1.62 (s, 3H), 1.50 (s, 9H), 1.33 (s, 3H).

[0419] Intermediate 5. (2R,3S,4R,5S)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-2-(((tert- butyldimethylsilyl)oxy)methyl)-3,4-dihydroxytetrahydrofuran-2-carbonitrile Intermediate 6. (2R,3S,4S,5S)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-2-(((tert- butyldimethylsilyl)oxy)methyl)-2-cyanotetrahydrofuran-3,4-diyl bis(2-methylpropanoate) Intermediate 7. (2R,3S,4S,5S)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-2-cyano- 2-(hydroxymethyl)tetrahydrofuran-3,4-diyl bis(2-methylpropanoate)

[0420]

[0421] This product can be prepared according to WO 2015 / 069939. For example, pages 127-138 of WO 2015 / 069939 provide a method for preparing the compound identified as Compound 14k in WO 2015 / 069939.

[0422] ​ ​

[0423]

[0424] Intermediate 3 (8.41 g, 18.87 mmol) was dissolved in THF (100 mL). TBAF 1.0 M in THF (28.31 mL, 28.31 mmol) was added in one portion at ambient temperature. Stirred at ambient temperature for 10 min. Reaction was determined to be complete by LCMS. The reaction mixture was quenched with water and the organics were removed under reduced pressure. The crude product was partitioned between EtOAc and water. The layers were separated and the aqueous phase was washed with EtOAc. The organics were combined and dried over sodium sulfate. The solids were filtered off and the solvent was removed under reduced pressure. The crude product was purified by silica gel chromatography (120 g column, 0-10% CH3OH in CH2Cl2) to give the product. LC / MS: t = 0.76 min, MS m / z = 332.14 [M+l]; LC system: Thermo Accela 1250 UHPLC. MS system: Thermo LCQ Fleet; Column: Kinetex 2.6 μ XB-C18 100A, 50 x 3.00 mm. Solvents: Acetonitrile with 0.1% formic acid, water with 0.1% formic acid. Gradient: 0 min - 2.4 min 2% - 100% ACN, 2.4 min - 2.80 min 100% ACN, 2.8 min - 2.85 min 100% - 2% ACN, 2.85 min - 3.0 min 2% ACN, at a rate of 1.8 mL / min. R H NMR (400 MHz, DMSO-d6) δ 7.87 - 7.80 (m, 3H), 6.85 (d, J = 4.5 Hz, 1H), 6.82 (d, J = 4.5 Hz, 1H), 5.74 (t, J = 5.8 Hz, 1H), 5.52 (d, J = 4.2 Hz, 1H), 5.24 (dd, J = 6.8, 4.2 Hz, 1H), 4.92 (d, J = 6.8 Hz, 1H), 3.65 (dd, J = 6.1, 1.7 Hz, 2H), 1.61 (s, 3H), 1.33 (s, 3H). 1 H NMR (400 MHz, DMSO-d6) δ 7.87 - 7.80 (m, 3H), 6.85 (d, J = 4.5 Hz, 1H), 6.82 (d, J = 4.5 Hz, 1H), 5.74 (t, J = 5.8 Hz, 1H), 5.52 (d, J = 4.2 Hz, 1H), 5.24 (dd, J = 6.8, 4.2 Hz, 1H), 4.92 (d, J = 6.8 Hz, 1H), 3.65 (dd, J = 6.1, 1.7 Hz, 2H), 1.61 (s, 3H), 1.33 (s, 3H).

[0425] ​ ​

[0426]

[0427] Intermediate 1 (2 g, 6.18 mmol) was dissolved in 50 mL DMF, to this solution was added tert-butyldimethylsilyl chloride (1 g, 7 mmol) and imidazole (1.26 g, 19 mmol). The resulting mixture was stirred at RT for 2 h and the reaction was diluted with EtOAc, washed with NH4Cl solution, the organic solvent was evaporated and the residue was purified by silica gel column chromatography eluting with 0-100% EtOAc in hexane to get the product. LCMS: MS m / z = 406.36 [M+1], t R = 3.25 min; HPLC system: Agilent 1290 II; Column: Phenomenex Kinetex C18, 2.6u 110A, 100 x 4.6 mm; Solvents: A: Water with 0.1% TFA, B: Acetonitrile with 0.1% TFA; Gradient: 2-98% B, 8.5 min gradient, at a rate of 1.5 mL / min. R = 3.25 min; HPLC system: Agilent 1290 II; Column: Phenomenex Kinetex C18, 2.6u 110A, 100 x 4.6 mm; Solvents: A: Water with 0.1% TFA, B: Acetonitrile with 0.1% TFA; Gradient: 2-98% B, 8.5 min gradient, at a rate of 1.5 mL / min.

[0428] ​ ​

[0429]

[0430] Intermediate 5 (1.8 g, 4.44 mmol) was dissolved in 15 mL THF, to this solution was added isobutyric anhydride (1.54 g, 9.8 mmol) and DMAP (179 mg, 1.45 mmol). The resulting mixture was stirred at RT for 5 min and the reaction was quenched with MeOH and then diluted with EtOAc, washed with brine, the organic solvent was dried over Na2S04and evaporated under vacuum. The residue was purified by silica gel column chromatography eluting with 0-100% EtOAc in hexane to get the product. LCMS: MS m / z = 546.16 [M+1], t R= 1.92 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Chromatography column: Kinetex 2.6 μ XB-C18 100 A, 50 x 4.6 mm; Solvents: Acetonitrile with 0.1% acetic acid, water with 0.1% acetic acid; Gradient: 0 min - 2.0 min 2% - 100% acetonitrile, 2.0 min - 3.05 min 100% acetonitrile, 3.05 min - 3.2 min 100% - 2% acetonitrile, 3.2 min - 3.5 min 2% ACN, at a rate of 2 μL / min. HPLC: t R = 3.88 min; HPLC system: Agilent 1290 II; Chromatography column: Phenomenex Kinetex C18, 2.6u 110 A, 100 x 4.6 mm; Solvents: A: water with 0.1% TFA, B: acetonitrile with 0.1% TFA; Gradient: 2% - 98% B, 8.5 min gradient, at a rate of 1.5 mL / min.

[0431] ​ ​

[0432]

[0433] Intermediate 6 (3.2 g, 5.86 mmol) was dissolved in 25 mL THF in a 100 mL plastic bottle to which HF-pyridine (10 g, 0.35 mmol) was added. The resulting mixture was stirred at RT for 3 h and the reaction was quenched with NaHC03, then diluted with EtOAc, washed with brine, the organic solvent was dried over Na2S04and evaporated under vacuum. The residue was purified by silica gel column chromatography eluting with 0% - 100% EtOAc in hexanes to give the product. 1 H NMR (400 MHz, Acetonitrile-d3) δ 7.90 (s, 1H), 6.83 - 6.74 (m, 2H), 6.33 (s, 2H), 5.84 - 5.74 (m, 2H), 5.62 (d, J = 5.4 Hz, 1H), 4.31 (dd, J = 8.4, 5.2 Hz, 1H), 3.94 (dd, J = 12.2, 5.0 Hz, 1H), 3.87 (dd, J = 12.2, 8.4 Hz, 1H), 2.70 (hept, J = 7.0 Hz, 1H), 2.56 (hept, J = 7.0 Hz, 1H), 1.28 - 1.17 (m, 6H), 1.12 (dd, J = 15.1, 7.0 Hz, 6H). LCMS: MS m / z = 432.24 [M+l], t R= 1.47 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Chromatography column: Kinetex 2.6 μ XB-C18 100 A, 50 x 4.6 mm; Solvents: Acetonitrile with 0.1% acetic acid, water with 0.1% acetic acid; Gradient: 0 min - 2.0 min 2% - 100% acetonitrile, 2.0 min - 3.05 min 100% acetonitrile, 3.05 min - 3.2 min 100% - 2% acetonitrile, 3.2 min - 3.5 min 2% ACN, at a rate of 2 μL / min. HPLC: t R = 2.74 min; HPLC system: Agilent 1290 II; Chromatography column: Phenomenex Kinetex C18, 2.6u 110 A, 100 x 4.6 mm; Solvents: A: water with 0.1% TFA, B: acetonitrile with 0.1% TFA; Gradient: 2% - 98% B, 8.5 min gradient, at a rate of 1.5 mL / min.

[0434] Intermediate 8. L-alanine cyclopentyl ester HC1 salt

[0435]

[0436] To a mixture of (tert-butoxycarbonyl)-L-alanine (3.95 g, 20.9 mmol), cyclopentanol (1.5 g, 17.4 mmol) and l-ethyl-3-(3-dimethylaminopropyl)carbodiimide HC1 salt (EDCI) (3.5 g, 22.6 mmol) in acetonitrile (100 mL) was added 4-(dimethylamino)pyridine (DMAP, 3.2 g, 26.1 mmol). The mixture was then stirred at room temperature for 2 hours, then the reaction mixture was diluted with EtOAc, washed with brine, the organic solvent was dried over sodium sulfate, then concentrated in vacuo. The obtained residue was purified by silica gel chromatography eluting with 0% - 100% ethyl acetate in hexanes to give an intermediate which was dissolved in 10 mL DCM to which was added 4N HC1 in dioxane (3 mL). The reaction mixture was stirred at RT for 30 min, then the solvent was evaporated and the residue was dried under high vacuum to give the crude product. 1 H NMR (400 MHz, Chloroform-d) δ 8.75 - 8.42 (m, 2H), 5.20 (tt, J = 5.6, 2.5 Hz, 1H), 4.22 - 4.07 (m, 1H), 1.87 - 1.58 (m, 8H), 1.54 (dd, J = 12.6, 7.2 Hz, 3H).

[0437] Intermediate 9. L-alanine cyclopropyl ester HC1 salt

[0438]

[0439] To a mixture of (tert-butoxycarbonyl)-L-alanine (5.86 g, 31 mmol), cyclopropanol (1.5 g, 25.8 mmol) and l-ethyl-3-(3-dimethylaminopropyl)carbodiimide HC1 salt (EDCI) (5.2 g, 33.6 mmol) in acetonitrile (100 mL) was added 4-(dimethylamino)pyridine (DMAP, 4.7 g, 38.7 mmol). The mixture was then stirred at room temperature for 2 hours, then the reaction mixture was diluted with EtOAc, washed with brine, the organic solvent was dried over sodium sulfate, then concentrated in vacuo. The obtained residue was purified by silica gel chromatography eluting with 0% - 100% ethyl acetate in hexanes to give an intermediate which was dissolved in 10 mL DCM to which was added 4N HC1 in dioxane (3 mL). The reaction mixture was stirred at RT for 30 min, then the solvent was evaporated and the residue was dried under high vacuum to give the crude product. 1 H NMR (400 MHz, Chloroform-d) δ 8.68 (s, 2H), 4.22 (tt, J = 6.3, 3.2 Hz, 1H), 1.68 (d, J = 7.3 Hz, 3H), 1.42 (s, 1H), 0.86 - 0.69 (m, 2H), 0.70 (dd, J = 7.1, 3.6 Hz, 2H).

[0440] Intermediate 10. Acetals 1 and 2: 1,1-dimethoxy-N,N-dimethylmethanamine and 1- (dimethoxymethyl)-4-methylpiperazine Intermediate 11. (S)-2-aminopropionic acid cyclohexyl ester HC1 salt

[0441]

[0442] A mixture of N-methylpiperazine (1.5 mL, 15.93 mmol) and DMF-dimethyl acetal (1 mL, 7.50 mmol) was heated in a sealed tube at 100 °C for 3 days, concentrated under high vacuum at 60 °C to remove excess N-methylpiperazine, then used in the next reaction. The product was a mixture of acetal 1 and acetal 2 in about 1 :2 ratio based on the product composition of the next reaction.

[0443] Intermediate 12. (S)-2-((tert-butoxycarbonyl)amino)-4-methylpentanoic acid 2- ethylbutyl ester

[0444]

[0445] To a mixture of L-alanine (5 g, 56.12 mmol) and cyclohexanol (56 g, 561 mmol) was added TMSC1 (20 mL). The resulting mixture was stirred at about 70 °C for about 15 hours and concentrated at about 80 °C under vacuum, co-evaporated with toluene, dissolved in hexanes and stirred at about room temperature during which time solids precipitated. The solids were collected by filtration, the filter cake was washed several times with 5% EtOAc in hexanes and dried under high vacuum for about 15 hours to give the product. 1 H NMR (400 MHz, Chloroform-d) δ 8.76 (s, 3H), 4.85 (tt, J = 8.7, 3.8 Hz, 1H), 4.17 (p, J = 6.5 Hz, 1H), 1.84 (dd, J = 9.9, 5.5 Hz, 2H), 1.70 (d, J = 7.3 Hz, 5H), 1.57 - 1.42 (m, 3H), 1.32 (dddd, J = 20.3, 12.8, 9.9, 6.4, 3.1 Hz, 3H).

[0446] Intermediate 13. (S)-2-amino-4-methylpentanoic acid 2-ethylbutyl ester HC1 salt

[0447]

[0448] (S)-2-((tert-Butoxycarbonyl)amino)-4-methylpentanoic acid (1.09 g, 4.71 mmol) was dissolved in acetonitrile (10 mL), 2-ethyl-1-butanol (2.88 mL, 23.56 mmol) was added followed by EDCI (878 mg, 5.66 mmol) and DMAP (863 mg, 7.07 mmol) in one portion. Stirred at room temperature overnight. Concentrated and diluted with CH2Cl2. Purified by silica gel chromatography 0% - 40% EtOAc in hexanes to give the product. 1 H NMR (400 MHz, DMSO-d6) δ 7.19 (d, J = 8.7 Hz, 1H), 4.00 - 3.84 (m, 3H), 1.67 - 1.22 (m, 17H), 0.91 - 0.80 (m, 12H).

[0449] Intermediate 14. 2-(benzyloxy)-2-methylpropyl ((4-nitrophenoxy)(phenoxy) phosphoryl)-L-alaninate

[0450]

[0451] (S)-2-((tert-Butoxycarbonyl)amino)-4-methylpentanoic acid 2-ethyl butyl ester was dissolved in CH2Cl2(10 mL) and 4N HCI in dioxane (10 mL, 40 mmol). Stirred at ambient temperature for 1 hour. Concentrated under reduced pressure and co-evaporated with hexanes. Left to sit under high vacuum for 1 hour, the product was used as is in the next step without purification. 1H NMR (400 MHz, DMSO-d6) δ 8.43 (s, 3H), 4.08 (d, J = 5.6 Hz, 2H), 3.92 (m, 1H), 1.69 (m, 1H), 1.61 (m, 2H), 1.47 (m, 1H), 1.34 (m, 4H), 0.83 (m, 12H).

[0452] Intermediate 15. ((4-nitrophenoxy)(phenoxy)phosphoryl)-L-alanine cyclobutyl methyl ester Intermediate 16. 2-ethylbutyl ((benzyloxy)(4-nitrophenoxy)phosphoryl)-L-alaninate

[0453]

[0454] To a mixture of Boc-L-alanine (1.26 g, 6.66 mmol), 2-benzyloxy-2-methylpropanol (1.0 g, 5.55 mmol), and EDCI (1.12 g, 7.21 mmol) in acetonitrile (20 mL) was added DMAP (2.04 g, 8.32 mmol). The mixture was then stirred at room temperature for 2 hours, then diluted with EtOAc, washed with brine, dried over sodium sulfate, and concentrated in vacuo. The obtained residue was purified by silica gel chromatography (0% to 60% EtOAc in hexanes) to give Boc-L-alanine propyl ester, which was dissolved in DCM (10 mL) and 4N HCl in dioxane (5.5 mL, 22.19 mmol) was added at room temperature. The resulting mixture was stirred at room temperature for 2 hours, concentrated in vacuo, redissolved in ACN (10 mL), and lyophilized overnight to give the product. 1 H NMR (400 MHz, Chloroform-d) δ 8.82 (s, 3H), 7.42 - 7.07 (m, 5H), 4.44 (s, 2H), 4.24 (m, 2H), 4.08 (d, J = 11.2 Hz, 1H), 1.70 (d, J = 7.0 Hz, 3H), 1.28 (d, J = 2.4 Hz, 6H). LCMS m / z = 251.97 (free base M+H), t R = 0.85 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Phenomenex Kinetex 2.6 μ XB-C18 100A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% acetonitrile, 1.8 min - 1.85 min 100% - 2% acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min.

[0455]

[0456] 2-(benzyloxy)-2-methylpropyl ((4-nitrophenoxy)(phenoxy) phosphoryl)-L-alaninate. To a solution of 2-(benzyloxy)-2-methylpropyl L-alaninate HC1 salt (832 mg, 2.89 mmol) in DCM (20 mL) was added phenyl dichlorophosphate (0.43 mL, 2.89 mmol) in one portion at -78 °C and triethylamine (0.80 mL, 5.76 mmol) was added dropwise over 5 min at -78 °C. After removing the dry ice bath, the resulting mixture was stirred for 30 min and cooled to -78 °C, and p-nitrophenol (402 mg, 2.89 mmol) was added in one portion and triethylamine (0.40 mL, 2.89 mmol) was added dropwise over 5 min at -78 °C. After removing the dry ice bath, the resulting mixture was stirred for 50 min, then diluted with DCM, washed with brine, concentrated in vacuo, and the resulting residue was purified by silica gel column chromatography (0% to 60% EtOAc in hexanes) to give the product. 1 HNMR (400 MHz, Chloroform-d) δ 8.23 - 8.13 (m, 2H), 7.41 - 7.27 (m, 3H), 7.28 - 7.14 (m, 4H), 4.45 (m, 2H), 4.27 - 4.15 (m, 2H), 4.07 (m, 1H), 3.89 (m, 1H), 1.41 (m, 3H), 1.27 (m, 6H). 31 P NMR (162 MHz, Chloroform-d) δ -3.10, -3.18. LCMS m / z = 528.78 (M+H), t R = 1.70 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Phenomenex Kinetex 2.6 μ XB-C18 100 A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% acetonitrile, 1.8 min - 1.85 min 100% - 2% acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min.

[0457] Intermediate 17. ((S)-(4-nitrophenoxy)(phenoxy)(phosphoryl)-L-alanine 2-ethylbutyl ester

[0458]

[0459] L-alanine cyclobutylmethyl ester-HCl (1.2 g, 7.16 mmol) was suspended in methylene chloride (10 mL), cooled to -78 °C, and phenyl dichlorophosphate (1.07 mL, 7.16 mmol) was added rapidly. Triethylamine (2.0 mL, 14.32 mmol) was added over 60 min at -78 °C, and the resulting mixture was stirred at room temperature for 3 h. The reaction mixture was cooled to 0 °C and 4-nitrophenol (996 mg, 7.16 mmol) was added in one portion. Then triethylamine (1.0 mL, 7.16 mmol) was added over 60 min. The mixture was then stirred at room temperature for 3 h, filtered, the filtrate was concentrated to one third volume, and filtered again. The filtrate was concentrated, and the residue was purified by silica gel column chromatography (0% to 35% EtOAc in hexanes) to give the product. 1 H NMR (400 MHz, Chloroform-d) δ 8.28 - 8.16 (m, 2H), 7.45 - 7.32 (m, 4H), 7.29 - 7.16 (m, 3H), 4.23 - 4.01 (m, 3H), 3.95 - 3.83 (m, 1H), 2.59 (m, 1H), 2.03 (m, 2H), 1.98 - 1.80 (m, 2H), 1.73 (m, 2H), 1.42 (d, J=3.2 Hz, 1.5H), 1.40 (d, J=3.3 Hz, 1.5H). 31 P NMR (162 MHz, Chloroform-d) δ -3.06, -3.11.

[0460] Intermediate 18. ((S)-(4-nitrophenoxy)(phenoxy)phosphoryl)-L-alanine isopropyl ester

[0461]

[0462] Under an argon atmosphere, 4-nitrophenyl phosphorodichloridate (2.00 g, 7.81 mmol) and triethylamine (2.18 mL, 15.6 mmol) were added sequentially to a suspension of 2-ethylbutyl L-alaninate hydrochloride (1.091 g, 18.9 mmol) in dichloromethane (23 mL) at 0 °C. After 1 h, benzyl alcohol (0.810 mL, 7.81 mmol) and triethylamine (1.09 mL, 7.81 mmol) were added sequentially at 0 °C, and the resulting mixture was allowed to warm to rt. After 1 h, the reaction mixture was diluted with dichloromethane (50 mL), washed with saturated aqueous sodium bicarbonate (50 mL) and brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude residue was purified by silica gel chromatography eluting with 0% - 100% ethyl acetate in hexanes to give the product. 1H NMR (400 MHz, Chloroform-d1) δ 8.30 - 8.07 (m, 2H), 7.42 - 7.28 (m, 7H), 5.18 - 5.09 (m, 2H), 4.70 (s, 1H), 4.08 - 3.95 (m, 2H), 3.68 (q, J = 9.4 Hz, 1H), 1.55 - 1.18 (m, 8H), 0.87 (t, J = 7.4 Hz, 6H). 31 P NMR (162 MHz, Chloroform-d1) δ 2.32 (s), 2.28 (s). LCMS: MS m / z = 463.00 [M-1], t R = 1.56 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Kinetex 2.6 μ XB-C18 100A, 50 x 4.6 mm; Solvents: Acetonitrile with 0.1% Acetic acid, Water with 0.1% Acetic acid; Gradient: 0 min - 2.0 min 2% - 100% Acetonitrile, 2.0 min - 3.05 min 100% Acetonitrile, 3.05 min - 3.2 min 100% - 2% Acetonitrile, 3.2 min - 3.5 min 2% ACN, at a rate of 2 μL / min.

[0463] Intermediate 19. ((S)-(4-nitrophenoxy)(phenoxy)phosphoryl)-L-alanine ethyl ester

[0464]

[0465] Prepared as described in WO 2016 / 069825.

[0466] Intermediate 20. ((4-nitrophenoxy)(phenoxy)phosphoryl)-L-alanine cyclopropyl methyl ester

[0467]

[0468] Prepared as described in Cho et al., J. Med. Chem. 2014, 57, 1812-1825.

[0469] Intermediate 21. 2-(((4-nitrophenoxy)(phenoxy)phosphoryl)amino)ethyl neopentanoate

[0470]

[0471] Prepared as described in US20120009147A1.

[0472] Intermediate 22. (2S)-tetrahydro-2H-pyran-4-yl 2-(((4-nitrophenoxy)(phenoxy) phosphoryl)amino)propanoate

[0473]

[0474] L-alanine cyclopropylmethyl ester-HCl (1.0 g, 5.57 mmol) was suspended in methylene chloride (10 mL), cooled to -78 °C, and phenyl dichlorophosphate (0.83 mL, 5.57 mmol) was added rapidly. A solution of triethylamine (1.54 mL, 11.13 mmol) in DCM (1.5 mL) was added at -78 °C over 30 min and stirred for 30 min. 4-nitrophenol (774 mg, 5.57 mmol) was added in one portion at -78 °C. Then a solution of triethylamine (0.77 mL, 7.16 mmol) in DCM (2 mL) was added over 30 min. The mixture was then stirred at the same temperature for 30 min, washed with water, saturated Na2C03 solution and brine, dried over sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column chromatography (0% to 20% EtOAc in hexanes) to give the product. 1 HNMR (400 MHz, Chloroform-d) δ 8.22 (m, 2H), 7.58 - 7.29 (m, 4H), 7.32 - 7.14 (m, 3H), 4.25 - 4.07 (m, 1H), 4.07 - 3.80 (m, 3H), 1.44 (d, J = 2.9 Hz, 1.5H), 1.42 (d, J = 2.9 Hz, 1.5H), 1.26 - 1.01 (m, 1H), 0.66 - 0.49 (m, 2H), 0.42 - 0.15 (m, 2H). 31 P NMR (162 MHz, Chloroform-d) δ -3.07, -3.11. MS m / z = 420.97.

[0475] Intermediate 23. (S)-1-methylpyrrolidin-3-yl ((4-nitrophenoxy)(phenoxy)phosphoryl)- L-alaninate

[0476]

[0477] 2-Aminoethyl pivalate hydrochloride. At RT, pivaloyl chloride (3.82 mL, 31.0 mmol) was added to a solution of tert-butyl (2-hydroxyethyl)carbamate (4.8 mL, 31.0 mmol) and diisopropylethylamine (5.4 mL, 31.0 mmol) in dichloromethane (150 mL). After 4 h, the resulting mixture was washed with saturated aqueous sodium bicarbonate solution (150 mL) and brine (150 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude colorless oil was dissolved in a solution of hydrochloric acid in dioxane (4 M, 50 mL) and stirred at RT, slowly precipitating a white solid from the solution. After 3 h, the solid was collected by vacuum filtration to give the product. 1 H NMR (400 MHz, CD3OD) δ 4.32 - 4.25 (m, 2H), 3.26 (t, J = 5.4 Hz, 2H), 1.23 (s, 9H).

[0478]

[0479] 2-(((4-nitrophenoxy)(phenoxy)phosphoryl)amino)ethyl neopentanoate. To a solution of 2-aminoethyl neopentanoate hydrochloride (0.861 g, 4.74 mmol) and phenyl dichlorophosphate (0.705 mL, 4.74 mmol) in dichloromethane (23 mL) was added triethylamine (1.2 mL, 9.4 mmol) under an argon atmosphere at 0 °C. The resulting mixture was allowed to warm to RT and stirred for 1.5 h. Then 4-nitrophenol (660 mg, 4.74 mmol) and triethylamine (0.66 mL, 4.7 mmol) were added. After 1 h, the reaction mixture was diluted with dichloromethane (50 mL) and the resulting mixture was washed with saturated aqueous sodium bicarbonate (50 mL) and brine (50 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude residue was purified via SiO2column chromatography (40 g SiO2Combiflash HP Gold column, 0% - 100% ethyl acetate / hexanes) to give the product. 1 H NMR (400 MHz, CDC13) δ 8.23 (d, J = 9.2 Hz, 2H), 7.47 - 7.31 (m, 4H), 7.29 - 7.16 (m, 3H), 4.18 - 4.06 (m, 2H), 3.45 - 3.31 (m, 2H), 1.17 (s, 9H). 31 P NMR (162 MHz, DMSO-de) δ -1.48 (s). MS m / z = 422.95 [M+l].

[0480] Intermediate 24. (R)-1-methylpyrrolidin-3-yl ((4-nitrophenoxy)(phenoxy)phosphoryl)- L-alaninate Intermediate 25. (2S)-cyclohexyl 2-(((4-nitrophenoxy)(phenoxy)phosphoryl)amino) propanoate

[0481]

[0482] (S)-tetrahydro-2H-pyran-4-yl 2-aminopropionate hydrochloride. To a mixture of L-alanine (500 mg, 5.61 mmol) and tetrahydro-2H-pyran-4-ol (5 g, 49.0 mmol) was added TMSCl (2 mL). The resulting mixture was stirred at 70 °C for 15 h and concentrated in vacuo, and the resulting solid was triturated with 5% EtOAc in hexanes, filtered, washed with 5% EtOAc in hexanes several times, and dried under high vacuum for 15 h to give the product which was used in the next reaction without any characterization.

[0483]

[0484] (2S)-tetrahydro-2H-pyran-4-yl 2-(((4-nitrophenoxy)(phenoxy)phosphoryl)amino)propanoate. (S)-tetrahydro-2H-pyran-4-yl 2-aminopropanoate hydrochloride (1.33 g, 6.34 mmol) was dissolved in methylene chloride (15 mL), cooled to -78 °C, and phenyl dichlorophosphate (1.137 mL, 7.61 mmol) was added rapidly. Triethylamine (2.2 mL, 15.2 mmol) was added over 30 min at -78 °C, and the resulting mixture was stirred at -78 °C for 30 min. Then 4-nitrophenol (882 mg, 6.34 mmol) was added in one portion over 30 min at -78 °C and triethylamine (1.1 mL, 7.61 mmol) was added. The mixture was stirred at -78 °C for 30 min, washed with water twice, washed with brine, dried over sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column chromatography (0% to 70% EtOAc in hexanes) to give the product. 1 H NMR (400 MHz, Chloroform-d) δ 8.22 (m, 2H), 7.49 - 7.06 (m, 7H), 4.95 (m, 1H), 4.14 (m, 1H), 4.07 - 3.80 (m, 3H), 3.52 (m, 2H), 1.95 - 1.81 (m, 2H), 1.64 m, 2H), 1.42 (m, 3H). 31 PNMR (162 MHz, Chloroform-d) δ -3.09, -3.13. MS m / z = 451 (M+H) + .

[0485] Intermediate 26. 4-(((2S)-2-(((4-nitrophenoxy)(phenoxy)phosphoryl)amino)propanoyl) oxy)piperidine-1-carboxylate Intermediate 27. trans-4-(trifluoromethyl)cyclohexyl ((4-nitrophenoxy)(phenoxy) phosphoryl)-L-alaninate

[0486]

[0487] (S)-1 -methylpyrrolidin-3-yl (tert-butoxycarbonyl)-L-alaninate. Boc-L-alanine (2.1 g, 11 mmol) and (R)-3-hydroxy-1 -methylpyrrolidine (1.1 mL, 10 mmol) were dissolved in dry THF (20 mL). Triphenylphosphine (3.4 g, 13 mmol) was added in one portion. Diisopropyl azodicarboxylate (2.4 mL, 12 mmol) was added dropwise. The reaction was stirred for 2 hours. More diisopropyl azodicarboxylate (240 uL, 1.2 mmol) was added dropwise and the reaction was stirred for 16 hours. The reaction was diluted with EtOAc (10 mL) and washed with saturated aqueous sodium bicarbonate (10 mL). The organics were then extracted with 5% aqueous citric acid (30 mL). The citric acid extract was washed with EtOAc (2 x 5 mL). The citric acid fraction was basified to pH 9 with 1 N aqueous NaOH and extracted with EtOAc (2 x 10 mL). The organic extracts were combined, dried over anhydrous sodium sulfate, then concentrated under reduced pressure to give the product. 1 H NMR (400 MHz, Chloroform-d) δ 5.24 (m, 1H), 5.01 (m, 1H), 4.27 (m, 1H), 2.88 - 2.69 (m, 2H), 2.64 (m, 1H), 2.37 (s, 3H), 2.29 (m, 1H), 1.96 - 1.80 (m, 1H), 1.44 (s, 9H), 1.37 (d, J=7.2 Hz, 3H).

[0488]

[0489] (S)-1 -methylpyrrolidin-3-yl ((4-nitrophenoxy)(phenoxy)phosphoryl)-L-alaninate. (S)-1 -methylpyrrolidin-3-yl (tert-butoxycarbonyl)-L-alaninate (545 mg, 2 mmol) was mixed with 10 mL of 4N HCI in dioxane and stirred for 1 hour. The reaction was concentrated under reduced pressure to give a foam which was mixed with 20 mL of dry DCM and stirred in an ice bath under atmospheric nitrogen. Phenyl dichlorophosphate (298 uL, 2 mmol) was added to the reaction in one portion. The reaction was stirred for 15 min. Triethylamine (837 uL, 6 mmol) was added dropwise to the reaction. The reaction was stirred for 1 hour. Triethylamine (279 uL, 2 mmol) was added dropwise to the reaction which was then stirred for 30 min. p-Nitrophenol (250 mg, 1.8 mmol) was added in one portion. The reaction mixture was stirred for 16 hours. The reaction was diluted with DCM (20 mL) and washed with water (5 x 20 mL). The organics were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude residue was purified via Si02column chromatography (12 g Si02Combiflash HP Gold column, 0% - 10% methanol / DCM). Fractions were combined and concentrated under reduced pressure to give the product. 1 H NMR (400 MHz, Chloroform-d) δ 8.28 - 8.15 (m, 2H), 7.46 - 7.28 (m, 4H), 7.28 - 7.13 (m, 3H), 5.17 (m, 1H), 4.21 - 4.04 (m, 1H), 4.01 - 3.85 (m, 1H), 2.81 (m, 1H), 2.70 - 2.55 (m, 2H), 2.35 (s, 3H), 2.33 - 2.21 (m, 2H), 1.84 - 1.70 (m, 1H), 1.39 (m, 3H). 31 P NMR (162 MHz, Chloroform-d) δ -3.16, -3.21. LCMS: MS m / z = 450.3 [M+1]; 448.1 [M-1], t R = 1.15 min; LC system: Thermo Dionex Ultimate 3000 UHPLC; Column: Phenomenex Kinetex 2.6 μ C18 100 A, 50 x 3 mm; Solvents: A: water with 0.1% acetic acid, B: acetonitrile with 0.1% acetic acid; Gradient: 0 min - 0.3 min 5% B, 0.3 min - 1.5 min 5% - 100% B, 1.5 min - 2 min 100% B, 2 min - 2.2 min 100% - 5% B, at a rate of 2 mL / min. HPLC: t R= 2.61 min; HPLC system: Agilent 1100 series; column: Phenomenex Gemini 5μ C18 110A, 50 x 4.6 mm; solvents: A: water with 0.1% TFA, B: acetonitrile with 0.1% TFA; gradient: 2-98% B in 5 min at a rate of 2 mL / min.

[0490] Intermediate 30. 1-methylpiperidin-4-yl ((4-nitrophenoxy)(phenoxy)phosphoryl)-L- alaninate Intermediate 31. (tetrahydro-2H-pyran-4-yl)methyl ((4-nitrophenoxy)(phenoxy) phosphoryl)-L-alaninate

[0491]

[0492] (R)-1 -methylpyrrolidin-3-yl (tert-butoxycarbonyl)-L-alaninate. Boc-L-alanine (5.2 g, 27.5 mmol) and (R)-3-hydroxy-1 -methylpyrrolidine (2.74 mL, 25 mmol) were dissolved in dry THF (25 mL). N,N'-diisopropylcarbodiimide (4.67 mL, 30 mmol) was added dropwise. The reaction was stirred for 2 hours. More N,N'-diisopropylcarbodiimide (467 uL, 3 mmol) was added dropwise and the reaction stirred for 2 hours. More N,N'-diisopropylcarbodiimide (467 uL, 3 mmol) was added dropwise and the reaction stirred for 16 hours.

[0493] The reaction was diluted with EtOAc (25 mL) and stirred for 10 min. The solids were filtered off and washed with a small amount of EtOAc. The filtrate was washed with saturated aqueous sodium bicarbonate (3 x 10 mL). The organics were then extracted with 5% aqueous citric acid (50 mL). The citric acid extract was washed with EtOAc (5 mL). The citric acid fraction was basified to pH 9 with 1 N aqueous NaOH and then extracted with EtOAc (3 x 15 mL). The organic extracts were combined, dried over anhydrous sodium sulfate, and then concentrated under reduced pressure to give the product. 1 H NMR (400 MHz, Chloroform-d) δ 5.28 - 5.18 (m, 1H), 5.02 (m, 1H), 4.28 (m, 1H), 2.84 - 2.75 (m, 1H), 2.69 (d, J=4.2 Hz, 2H), 2.36 (s, 3H), 2.34 - 2.22 (m, 2H), 1.87 - 1.76 (m, 1H), 1.44 (s, 9H), 1.37 (d, J=7.2 Hz, 3H).

[0494]

[0495] (R)-1 -methylpyrrolidin-3-yl ((4-nitrophenoxy)(phenoxy)phosphoryl)-L-alaninate. (R)-1 -methylpyrrolidin-3-yl (tert-butoxycarbonyl)-L-alaninate (3.9 g, 14.3 mmol) was mixed with 30 mL of 4N HCI in dioxane and stirred for 3 hours. The reaction was concentrated under reduced pressure to give a foam which was mixed with 30 mL of dry DCM and stirred in an ice bath under atmospheric nitrogen. Phenyl dichlorophosphate (2.34 mL, 15.75 mmol) was added to the reaction in one portion. The reaction was stirred for 15 min. Triethylamine (4.4 mL, 31.5 mmol) was mixed with dry DCM (5 mL) and added dropwise to the reaction. The reaction was stirred for 1 hour. Triethylamine (2.2 mL, 15.75 mmol) was mixed with dry DCM (3 mL) and added dropwise to the reaction. The reaction was stirred for 15 min. p-Nitrophenol (1.8 g, 12.87 mmol) was added in one portion. The reaction mixture was stirred for 2 hours.

[0496] The reaction was diluted with DCM (20 mL) and washed with aqueous sodium bicarbonate solution (3 x 20 mL). The organics were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude residue was purified via SiO2column chromatography (40 g SiO2Combiflash HP Gold column, 0% - 10% methanol / DCM). The fractions were combined and concentrated under reduced pressure to give the product. 1 H NMR (400 MHz, Chloroform-d) δ 8.29 - 8.15 (m, 2H), 7.48 - 7.29 (m, 4H), 7.29 - 7.13 (m, 3H), 5.20 (m, 1H), 4.21 - 4.07 (m, 1H), 3.99 (m, 1H), 2.86 (m, 1H), 2.70 (m, 1H), 2.63 (m, 1H), 2.37 (m, 3H), 2.35 - 2.21 (m, 2H), 1.86 - 1.73 (m, 1H), 1.40 (m, 3H). 31 P NMR (162 MHz, Chloroform-d) δ -3.12, -3.14. LCMS: MS m / z = 450.3 [M+1]; 448.1 [M-1], t R= 1.24 min; LC system: Thermo Dionex Ultimate 3000 UHPLC; column: Phenomenex Kinetex 2.6 μ C18 100A, 50 x 3 mm; solvents: A: water with 0.1 % acetic acid, B: acetonitrile with 0.1 % acetic acid; gradient: 0 min - 0.3 min 5 % B, 0.3 min - 1.5 min 5 % - 100 % B, 1.5 min - 2 min 100 % B, 2 min - 2.2 min 100 % - 5 % B, rate 2 mL / min. HPLC: t R = 2.63 min; HPLC system: Agilent 1100 series; column: Phenomenex Gemini 5 μ C18 110A, 50 x 4.6 mm; solvents: A: water with 0.1 % TFA, B: acetonitrile with 0.1 % TFA; gradient: 2 % - 98 % B in 5 min, rate 2 mL / min.

[0497] Intermediate 32. trans-4-(tert-butyl)cyclohexyl ((S)-(4-nitrophenoxy)(phenoxy) phosphoryl)-L-alaninate

[0498]

[0499] Intermediate 11 (3.4 g, 16.37 mmol) was dissolved in methylene chloride (45 mL), cooled to -78 °C, and phenyl phosphorodichloridate (2.45 mL, 16.37 mmol) was added rapidly. Triethylamine (4.54 mL, 32.74 mmol) was added over 60 min at -78 °C, followed by 4-nitrophenol (2277 mg, 16.37 mmol) in one portion. Triethylamine (2.27 mL, 16.37 mmol) was added over 60 min at -78 °C. The resulting mixture was stirred at -78 °C for 2 h, diluted with methylene chloride (100 mL), washed twice with water, washed with brine, dried over sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column chromatography (0 % to 20 % EtOAc in hexanes) to give the product. 1 HNMR (400 MHz, Chloroform-d) δ 8.22 (m, 2H), 7.46 - 7.30 (m, 4H), 7.29 - 7.09 (m, 3H), 4.76 (m, 1H), 4.20 - 4.02 (m, 1H), 3.92 (m, 1H), 1.87 - 1.64 (m, 4H), 1.54 (m, 2H), 1.46 - 1.18 (m, 7H). 31 P NMR (162 MHz, Chloroform-d) δ -2.94, -3.00. MS m / z = 449 (M+H) + .

[0500] Intermediate 33. ((1 r,4S)-4-(trifluoromethyl)cyclohexyl)methyl ((4-nitrophenoxy)(phenoxy) phosphoryl)-L-alaninate ​

[0501]

[0502] 4-((L-alanyl)oxy)piperidine-1 -carboxylic acid tert-butyl ester. To a mixture of ((benzyloxy)carbonyl)-L-alanine (1.26 g, 5.65 mmol), 4-hydroxypiperidine-1 -carboxylic acid tert-butyl ester (5.68 g, 28.22 mmol) and EDCI (1.05 g, 6.77 mmol) in acetonitrile (15 mL) was added DMAP (1.03 g, 8.47 mmol). The mixture was then stirred at room temperature for 15 hours, diluted with EtOAc, washed with brine, dried over sodium sulfate, and concentrated in vacuo. The obtained residue was purified by silica gel chromatography (0% to 100% EtOAc in hexanes) to give Cbz-L-alanine piperidyl ester which was dissolved in THF (10 mL) and 20% carbon supported palladium hydroxide (400 mg) was added. The resulting mixture was stirred under H2gas for 2 hours, filtered, and the filtrate was concentrated in vacuo. The obtained residue was dried under high vacuum to give the product. 1 H NMR (400 MHz, Chloroform-d) δ 4.95 (tt, J = 7.9, 3.8 Hz, 1H), 3.79 - 3.62 (m, 2H), 3.56 (q, J = 7.0 Hz, 1H), 3.25 (ddd, J = 13.6, 8.5, 3.7 Hz, 2H), 1.85 (ddd, J = 13.4, 6.4, 3.4 Hz, 2H), 1.73 (s, 2H), 1.62 (ddq, J = 12.7, 8.7, 4.3, 3.9 Hz, 2H), 1.46 (s, 9H), 1.34 (d, J = 7.0 Hz, 3H). MS m / z = 273 [M+H].

[0503]

[0504] 4-(((2S)-2-(((4-nitrophenoxy)(phenoxy)phosphoryl)amino)propanoyl)oxy)piperidine- 1 -carboxylic acid tert-butyl ester. Dissolve 4-((L-alaninyl)oxy)piperidine-1 -carboxylic acid tert-butyl ester (0.9 g, 3.31 mmol) in methylene chloride (10 mL), cool to -78 °C, and rapidly add phenyl dichlorophosphate (0.49 mL, 3.31 mmol). Add triethylamine (0.46 mL, 3.31 mmol) over 30 min at -78 °C, and add 4-nitrophenol (460 mg, 3.31 mmol) in one portion. Then add triethylamine (0.49 mL, 3.31 mmol) over 30 min at -78 °C. Stir the resulting mixture at -78 °C for 2 h, dilute with methylene chloride, wash twice with water, wash with brine, dry over sodium sulfate, and concentrate in vacuo. Purify the residue by silica gel column chromatography (0% to 70% EtOAc in hexanes) to give the product. 1 H NMR (400 MHz, Chloroform-d) δ 8.23 (m, 2H), 7.42 - 7.31 (m, 4H), 7.25 - 7.16 (m, 3H), 4.93 (m, 1H), 4.26 - 4.03 (m, 1H), 3.85 (m, 1H), 3.75 - 3.56 (m, 2H), 3.21 (m, 2H), 1.91 - 1.75 (m, 2H), 1.66 - 1.48 (m, 2H), 1.46 (s, 9H), 1.44 - 1.38 (m, 3H). 31 P NMR (162 MHz, Chloroform-d) δ -3.07, -3.13. MS m / z = 550 (M+H) + .

[0505] ​ ​

[0506]

[0507] Trans-4-(trifluoromethyl)cyclohexyl L-alaninate. The product was prepared in a similar manner as described for Intermediate 26 from Cbz-L-alanine (900 mg, 4.03 mmol) and trans-4-(trifluoromethyl)cyclohexan-1-ol (1.02 g, 6.05 mmol). MS m / z = 240 [M+H].

[0508]

[0509] Trans-4-(trifluoromethyl)cyclohexyl ((4-nitrophenoxy)(phenoxy)phosphoryl)-L-alaninate. The product as a mixture of isomers was prepared in a similar manner as described for Intermediate 25 from trans-4-(trifluoromethyl)cyclohexyl L-alaninate (974 mg, 4.07 mmol) (840 mg). 1 H NMR (400 MHz, Chloroform-d) δ 8.27 - 8.19 (m, 2H), 7.43 - 7.31 (m, 4H), 7.26 - 7.16 (m, 3H), 4.68 (m, 1H), 4.11 (m, 1H), 3.84 (m, 1H), 2.02 (m, 4H), 1.50 - 1.27 (m, 8H). 19 F NMR (377 MHz, Chloroform-d) δ -73.91 (d, J = 7.7 Hz). 31 P NMR (162 MHz, Chloroform-d) δ -3.08, -3.12. MS m / z = 517 [M+H].

[0510] The product was separated by Chiralpak SFC (Chiralpak IF 20 x 250 mm column, 30% isopropyl alcohol) to give Intermediate 28 and Intermediate 29:

[0511]

[0512] Intermediate 28. Trans-4-(trifluoromethyl)cyclohexyl ((R)-(4-nitrophenoxy)(phenoxy)phosphoryl)-L-alaninate. First eluting diastereomer of Intermediate 27: 1 H NMR (400 MHz, Chloroform-d) δ 8.22 (d, J = 9.1 Hz, 2H), 7.42 - 7.31 (m, 4H), 7.29 - 7.16 (m, 3H), 4.69 (tt, J = 10.7, 4.2 Hz, 1H), 4.19 - 4.04 (m, 1H), 3.90 (dd, J = 11.9, 9.5 Hz, 1H), 2.12 - 1.97 (m, 5H), 1.52 - 1.21 (m, 7H). 19 F NMR (376 MHz, Chloroform-d) δ -73.90 (d, J = 7.7 Hz). 31 P NMR (162 MHz, Chloroform-d) δ -3.07.

[0513] Intermediate 29. Trans 4-(trifluoromethyl)cyclohexyl ((S)-(4-nitrophenoxy)(phenoxy)phosphoryl)-L-alaninate. Second eluting diastereomer of Intermediate 27: 1H NMR (400 MHz, Chloroform-d) δ 8.21 (d, J = 9.08 Hz, 2H), 7.42 - 7.31 (m, 4H), 7.26 - 7.13 (m, 3H), 4.67 (tt, J = 10.8, 4.2 Hz, 1H), 4.11 (ddt, J = 15.8, 8.9, 7.1 Hz, 1H), 3.97 (dd, J = 12.0, 9.4 Hz, 1H), 2.07 - 1.91 (m, 5H), 1.51 - 1.19 (m, 7H). 19 F NMR (376 MHz, Chloroform-d) δ -73.90 (d, J = 7.9 Hz). 31 P NMR (162 MHz, Chloroform-d) δ -3.08.

[0514] ​

[0515]

[0516] 1 -Methylpiperidin-4-yl L-alaninate. To a mixture of N-Cbz-L-alanine (1.047 g, 4.688 mmol), 4-hydroxy-N-methylpiperidine (450 mg, 3.907 mmol) and EDCI (788 mg, 5.079 mmol) in acetonitrile (20 mL) was added DMAP (716 mg, 5.861 mmol). The mixture was then stirred at room temperature for 15 hours, then diluted with EtOAc, washed with brine, dried over sodium sulfate and concentrated in vacuo. The obtained residue was purified by silica gel chromatography (0% to 10% MeOH in DCM) to give Cbz-L-alanine 4-piperidinyl ester which was dissolved in THF (10 mL) and 20% Pd(OH)2(300 mg, 0.427 mmol) was added at room temperature. The resulting mixture was stirred under H2gas at room temperature for 2 hours, filtered, concentrated in vacuo, co-evaporated with DCM several times and dried under high vacuum overnight to give the product. 1 H NMR (400 MHz, Chloroform-d) δ 4.81 (td, J = 8.3, 7.7, 3.8 Hz, 1H), 3.52 (q, J = 7.0 Hz, 1H), 2.63 (s, 2H), 2.29 (s, 5H), 2.14 - 1.86 (m, 4H), 1.73 (ddt, J = 12.9, 8.8, 4.5 Hz, 2H), 1.32 (d, J = 7.0 Hz, 3H). LCMS: MS m / z = 187.09 [M+l]; t R= 0.12 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Phenomenex Kinetex 2.6μ XB-C18 100A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% acetonitrile, 1.8 min - 1.85 min 100% - 2% acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min.

[0517]

[0518] 1 -Methylpiperidin-4-yl ((4-nitrophenoxy)(phenoxy)phosphoryl)-L-alaninate. To a solution of 1 -methylpiperidin-4-yl L-alaninate (360 mg, 1.706 mmol) in DCM (10 mL) was added phenyl dichlorophosphate (0.255 mL, 1.706 mmol) in one portion at -78 °C, followed by a solution of triethylamine (0.24 mL, 1.706 mmol) in DCM (2.76 mL) over 30 min at -78 °C. After removing the dry ice bath, the resulting mixture was stirred for 30 min, then cooled to -78 °C again. p-Nitrophenol (0.237 g, 1.706 mmol) was added in one portion, and triethylamine (0.237 mL, 1.706 mmol) was added over 30 min at -78 °C. After removing the dry ice bath, the resulting mixture was stirred for 30 min, then diluted with EtOAc, washed with water and brine, concentrated in vacuo, and the resulting residue was purified by silica gel column chromatography (0% to 10% MeOH in DCM) to give the product. 1 HNMR (400 MHz, Chloroform-d) δ 8.28 - 8.15 (m, 2H), 7.36 (m, 4H), 7.25 - 7.17 (m, 3H), 4.80 (s, 1H), 4.19 - 4.04 (m, 1H), 3.93 (m, 1H), 2.64 (s, 2H), 2.31 (m, 5H), 1.90 (m, 2H), 1.78 - 1.67 (m, 2H), 1.47 - 1.33 (m, 3H). 31 P NMR (162 MHz, Chloroform-d) δ -3.04, -3.07. LCMS: MS m / z = 464.32 [M+l]; t R= 0.74 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Chromatography column: Phenomenex Kinetex 2.6 μ XB-C18 100A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% acetonitrile, 1.8 min - 1.85 min 100% - 2% acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min.

[0519] ​ ​

[0520]

[0521] (Tetrahydro-2H-pyran-4-yl)methyl ((benzyloxy)carbonyl)-L-alaninate. Cbz-L-Ala (446 mg, 2 mmol) was dissolved in anhydrous MeCN (10 mL). EDCI (422 mg, 2.2 mmol) was added in one portion and the reaction stirred for 15 min. Tetrahydropyran-4-methanol (279 uL, 2.4 mmol) was then added. DMAP (269 mg, 2.2 mmol) was then added in one portion. The reaction was stirred for 16 h.

[0522] The reaction was diluted with EtOAc (30 mL), washed with 5% aqueous citric acid (10 mL), followed by saturated aqueous sodium bicarbonate (10 mL), and finally brine (10 mL). The organics were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude residue was purified via SiO2column chromatography (12 g SiO2Combiflash HP Gold column, 0% - 80% ethyl acetate / hexanes). Fractions were combined and concentrated under reduced pressure to yield the product. 1 H NMR (400 MHz, Chloroform-d) δ 7.40 - 7.28 (m, 5H), 5.28 (d, J = 7.9 Hz, 1H), 5.11 (s, 2H), 4.39 (t, J = 7.4 Hz, 1H), 4.07 - 3.84 (m, 4H), 3.38 (t, J = 11.7 Hz, 2H), 1.92 (s, 1H), 1.68 - 1.50 (m, 3H), 1.39 (m, 4H).

[0523]

[0524] (Tetrahydro-2H-pyran-4-yl)methyl ((4-nitrophenoxy)(phenoxy)phosphoryl)-L-alaninate. (Tetrahydro-2H-pyran-4-yl)methyl ((benzyloxy)carbonyl)-L-alaninate (530 mg, 1.65 mmol) was dissolved in anhydrous THF (12 mL). 10% Pd / C Degussa type was added and the reaction mixture was stirred under atmospheric hydrogen for 2 hours. The catalyst was filtered and the filtrate used without purification.

[0525] Phosphorous dichloride (294 uL, 1.98 mmol) was dissolved in anhydrous DCM (10 mL) and stirred in an ice bath under atmospheric nitrogen. The above THF solution was added dropwise to the reaction and then stirred for 10 min. Triethylamine (300 uL, 2.15 mmol) was added dropwise and then stirred for 30 min. p-Nitrophenol (207 mg, 1.49 mmol) and triethylamine (300 uL, 2.15 mmol) were added. The ice bath was removed and the reaction mixture was stirred at RT for 14 hours.

[0526] The reaction was diluted with EtOAc (30 mL) and washed with 0.2M sodium carbonate solution (2 x 10 mL) followed by brine (10 mL). The organics were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude residue was purified by SiO2column chromatography (12g SiO2Combiflash HP Gold column, 0% - 50% ethyl acetate / hexane). Fractions were combined and concentrated under reduced pressure to give the product. 1 HNMR (400 MHz, Chloroform-d) δ 8.23 (d, J = 9.0 Hz, 2H), 7.45 - 7.30 (m, 4H), 7.30 - 7.16 (m, 3H), 4.23 - 4.07 (m, 2H), 3.97 (m, 4H), 3.85 (t, J = 10.5 Hz, 1H), 3.35 (t, J = 11.8 Hz, 2H), 1.99 - 1.79 (m, 1H), 1.56 (d, J = 8.4 Hz, 3H), 1.48 - 1.29 (m, 4H). 31 P NMR (162 MHz, Chloroform-d) δ -3.13 (s), -3.16 (s). MS m / z = 464.9 [M+1]; 463.1 [M-1].

[0527] ​ ​

[0528]

[0529] Trans-4-(tert-butyl)cyclohexyl ((4-nitrophenoxy)(phenoxy)phosphoryl)-L-alaninate. The product was prepared in a similar manner as described for Intermediate 25 from trans-4-(tert-butyl)cyclohexyl L-alaninate (420 mg, 1.85 mmol) as a mixture of isomers (520 mg). 1 H NMR (400 MHz, Chloroform-d) δ 4.65 (tt, J = 11.2, 4.5 Hz, 1H), 3.51 (q, J = 7.1 Hz, 1H), 2.07 - 1.93 (m, 2H), 1.87 - 1.73 (m, 4H), 1.40 - 1.23 (m, 4H), 1.19 - 0.94 (m, 4H), 0.85 (d, J = 2.6 Hz, 9H). MS m / z = 228 [M+H].

[0530]

[0531] Trans-4-(tert-butyl)cyclohexyl ((4-nitrophenoxy)(phenoxy)phosphoryl)-L-alaninate. The product was prepared in a similar manner as described for Intermediate 25 from trans-4-(tert-butyl)cyclohexyl L-alaninate (420 mg, 1.85 mmol) as a mixture of isomers (520 mg). 1 H NMR (400 MHz, Chloroform-d) δ 8.27 - 8.19 (m, 2H), 7.37 (m, 4H), 7.28 - 7.16 (m, 3H), 4.62 (m, 1H), 4.17 - 4.00 (m, 1H), 3.88 (m, 1H), 1.95 (m, 2H), 1.80 (m, 2H), 1.39 (m, 3H), 1.35 - 1.22 (m, 2H), 1.15 - 0.92 (m, 3H), 0.85 (s, 9H). 31 P NMR (162 MHz, Chloroform-d) δ -2.98, -3.04. MS m / z = 505 [M+H].

[0532] ​ ​

[0533]

[0534] ((1s,4s)-4-(trifluoromethyl)cyclohexyl)methanol. To an ice-cold solution of (1s,4s)-4-(trifluoromethyl)cyclohexanecarboxylic acid (3 g, 15.29 mmol) in anhydrous tetrahydrofuran (40 mL) was added lithium aluminum hydride (0.871 g, 22.94 mmol) in portions over 30 min. The reaction mixture was stirred at room temperature for 3 h. It was cooled to 0 °C and quenched with water (0.8 mL), 5 N aqueous sodium hydroxide solution (0.8 mL), and again with water (2.4 mL). The isolated solid was filtered off and the filtrate was diluted with ethyl acetate and saturated aqueous sodium bicarbonate solution. The organic layer was separated, washed with brine and dried over sodium sulfate. The ethyl acetate was filtered off and concentrated under reduced pressure to give the product. The obtained residue was dried under high vacuum for 1 h and used as such in the subsequent reaction. 1 H NMR (400 MHz, Chloroform-d) δ 3.47 (dd, J = 6.3, 1.9 Hz, 2H), 2.08 - 1.77 (m, 5H), 1.62 - 1.18 (m, 4H), 0.99 (qd, J = 13.0, 3.2 Hz, 2H). 19 F NMR (376 MHz, Chloroform-d) δ -74.33 (d, J = 8.2 Hz).

[0535]

[0536] ((1r,4S)-4-(trifluoromethyl)cyclohexyl)methyl (tert-butoxycarbonyl)-L-alaninate. The product (1.48 g) was prepared in a similar manner as described for Intermediate 12. 1 H NMR (400 MHz, Chloroform-d) δ 5.00 (s, 1H), 4.30 (s, 1H), 4.04 - 3.89 (m, 2H), 2.08 - 1.79 (m, 5H), 1.74 - 1.57 (m, 1H), 1.44 (s, 9H), 1.38 (d, J = 7.2 Hz, 3H), 1.30 (m, 2H), 1.12 - 0.93 (m, 2H). 19 F NMR (376 MHz, Chloroform-d) δ -74.38 (d, J = 7.8 Hz).

[0537]

[0538] (S)-1-oxo-1-(((1r,4S)-4-(trifluoromethyl)cyclohexyl)methoxy)propan-2- amine hydrochloride. The product (1.184 g) was prepared in a similar manner as described for Intermediate 13. 1H NMR (400 MHz, DMSO-d6) δ 8.55 (s, 3H), 4.17 - 3.88 (m, 3H), 2.21 (dtd, J = 12.2, 8.8, 3.3 Hz, 1H), 1.83 (ddd, J = 29.5, 13.4, 3.4 Hz, 4H), 1.63 (tdd, J = 11.9, 6.0, 3.3 Hz, 1H), 1.41 (d, J = 7.2 Hz, 3H), 1.32 - 0.93 (m, 4H). 19 F NMR (377 MHz, DMSO-d6) δ -72.84 (d, J = 8.8 Hz).

[0539]

[0540] ((1r,4S)-4-(trifluoromethyl)cyclohexyl)methyl ((4-nitrophenoxy)(phenoxy)phosphoryl)-L-alaninate. The product (1.4 g) was prepared in a similar manner as described for Intermediate 35. 1 H NMR (400 MHz, DMSO-d6) δ 8.37 - 8.22 (m, 2H), 7.56 - 7.31 (m, 4H), 7.30 - 7.14 (m, 2H), 6.72 (ddd, J = 13.7, 10.1, 8.6 Hz, 1H), 4.10 - 3.91 (m, 1H), 3.88 - 3.75 (m, 2H), 2.20 - 1.99 (m, 1H), 1.86 - 1.63 (m, 4H), 1.54 - 1.41 (m, 1H), 1.29 - 1.06 (m, 5H), 0.98 (td, J = 12.7, 3.2 Hz, 2H). MS m / z = 531.02 [M+l].

[0541] Intermediate 34. ((S)-(perfluorophenoxy)(pentoxy)phosphoryl)-L-alanine ethyl ester

[0542]

[0543] To a solution of L-alanine ethyl ester-HCl (631 mg, 2.465 mmol) in DCM (15 mL) was added phenyl phosphorodichloridate (0.368 mL, 2.465 mmol) in one portion at -78 °C and triethylamine (0.68 mL, 4.93 mmol) was added dropwise over 5 min at -78 °C. After removing the dry ice bath, the resulting mixture was stirred for 30 min and then cooled to -78 °C. Pentakisfluorophenol (454 mg, 2.465 mmol) was added in one portion and triethylamine (0.34 mL, 2.465 mmol) was added over 5 min at -78 °C. After removing the dry ice bath, the resulting mixture was stirred for 1 hour, then diluted with DCM, washed with brine, concentrated in vacuo and the resulting residue was purified by silica gel column chromatography (0% to 60% EtOAc in hexanes) to give a diastereomeric mixture to which diisopropyl ether (4 mL) was added. The suspension was sonicated and filtered. The filter cake was washed with diisopropyl ether (2 mL) and the filtrate was concentrated in vacuo to give a diastereomeric mixture as a white solid. 1 H NMR showed it to be a 3:1 mixture. Diisopropyl ether (5 mL) was added to the filter cake and the suspension was heated at 70 °C to a clear solution. After removing the hot bath, needle-like crystals started to form, after 10 min, the mixture was filtered and the filter cake was dried under high vacuum for 30 min to give the Sp isomer.

[0544] Diastereomeric mixture: 1 H NMR (400 MHz, Chloroform-d) δ 7.43 - 7.30 (m, 2 H), 7.32 - 7.17 (m, 3 H), 4.29 - 4.11 (m, 3 H), 3.94 (m, 1 H), 1.52 - 1.42 (m, 3 H), 1.28 (q, J = 7.0 Hz, 3 H).

[0545] Sp isomer: 1 H NMR (400 MHz, Acetonitrile-d3) δ 7.50 - 7.36 (m, 2 H), 7.32 - 7.21 (m, 3 H), 4.75 (t, J = 11.5 Hz, 1 H), 4.17 - 3.98 (m, 3 H), 1.37 (dd, J = 7.1, 1.1 Hz, 3 H), 1.22 (t, J = 7.1 Hz, 3 H). 31 P NMR (162 MHz, Acetonitrile-d3) δ -0.51. 19 F NMR (376 MHz, Acetonitrile-d3) δ -155.48 - -155.76 (m), -162.73 (td, J = 21.3, 3.7 Hz), -165.02 - -165.84 (m). LCMS m / z = 440.5 (M-ethyl+H), t R= 1.57 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Chromatography column: Phenomenex Kinetex 2.6 μ XB-C18 100A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% acetonitrile, 1.8 min - 1.85 min 100% - 2% acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min.

[0546] Intermediate 35. (2S)-2-ethylbutyl 2-cyclohexyl-2-(((4-nitrophenoxy)(pentoxy)phosphoryl)amino)acetate Intermediate 36. (l-(2,2,2-trifluoroethyl)piperidin-4-yl)methyl ((4-nitrophenoxy)(pentoxy)phosphoryl)alaninate

[0547]

[0548] (S)-2-ethylbutyl 2-amino-2-cyclohexylethanoate hydrochloride. L- cyclohexylglycine (0.90 g, 5.75 mmol) was dissolved in 2-ethyl-1-butanol (20 mL) and trimethylchlorosilane (1.31 mL, 10.30 mmol) was added in one portion. It was left in a preheated 60 °C oil bath for 16 h. Concentrated and co-evaporated with toluene 5 times in a 60 °C rotary evaporator bath. Left in high vacuum overnight to give the product. This material was used as is in the next step. 1 H NMR (400 MHz, DMSO-d6) δ 8.38 (s, 3H), 4.17 - 3.96 (m, 2H), 3.84 (d, J = 4.5 Hz, 1H), 1.90 - 1.40 (m, 5H), 1.41 - 0.88 (m, 11H), 0.83 (t, J = 7.3 Hz, 6H).

[0549]

[0550] (2S)-2-ethylbutyl 2-cyclohexyl-2-(((4-nitrophenoxy)(phenoxy)phosphoryl)amino)acetate. To a solution of (S)-2-ethylbutyl 2-amino-2-cyclohexylacetate hydrochloride (1.50 g, 5.39 mmol) and phenyl dichlorophosphate (0.803 mL, 5.39 mmol) in dichloromethane (50 mL) was added triethylamine (1.56 mL, 11.16 mmol) at 0 °C under an argon atmosphere. The resulting mixture was allowed to warm to RT and stirred for 1 h. Then 4-nitrophenol (713 mg, 5.13 mmol) and triethylamine (0.81 mL, 5.63 mmol) were added. After 2 h, the reaction mixture was diluted with Et2O (100 mL) and the solids were filtered off. The crude product was concentrated under reduced pressure and purified by silica gel chromatography (120 g SiO2Combiflash HP Gold column, 0% - 50% ethyl acetate / hexanes) followed by purification by reverse phase HPLC (20% - 100% ACN / water) without modifier to give the product. 1 H NMR (400 MHz, DMSO-d6) δ 8.28 (br d, J = 9.3 Hz, 2H), 7.55 - 7.28 (m, 4H), 7.28 - 7.01 (m, 3H), 6.61 - 6.52 (m, 1H), 3.85 (d, J = 4.0 Hz, 2H) 3.75 - 3.53 (m, 1H), 1.67 - 1.31 (m, 7H), 1.25 (m, 6H), 1.16 - 0.67 (m, 9H). LC / MS: t R = 1.48 min, MS m / z = 519.03 [M+1]; LC system: Thermo Accela 1250 UHPLC. MS system: Thermo LCQ Fleet; Column: Kinetex 2.6 μ XB-C18 100 A, 50 x 3.00 mm. Solvents: Acetonitrile with 0.1% formic acid, water with 0.1% formic acid. Gradient: 0 min - 2.4 min 2% - 100% ACN, 2.4 min - 2.80 min 100% ACN, 2.8 min - 2.85 min 100% - 2% ACN, 2.85 min - 3.0 min 2% ACN, at a rate of 1.8 mL / min.

[0551] Intermediate 37. (l-ethyl-3,3-difluoropiperidin-4-yl ((4-nitrophenoxy)(pentoxy)phosphoryl)-L-alanine ester Intermediate 38. 4-nitrophenyl-N,N'-ethyl L-alanine diaminophosphonate

[0552]

[0553] (1-(2,2,2-trifluoroethyl)piperidin-4-yl)methyl (tert-butoxycarbonyl)alaninate. The product (3.8 g) was prepared in a similar manner as described for Intermediate 12.1 H NMR (400 MHz, DMSO-d6) δ 7.25 (d, J = 7.4 Hz, 1H), 4.08 - 3.72 (m, 3H), 3.10 (q, J = 10.3 Hz, 2H), 2.88 (d, J = 11.0 Hz, 2H), 2.37 - 2.18 (m, 2H), 1.66 - 1.47 (m, 3H), 1.36 (s, 9H), 1.21 (d, J = 7.5 Hz, 5H). 19 F NMR (376 MHz, DMSO-d6) δ -68.52 (t, J = 10.3 Hz).

[0554]

[0555] (1-(2,2,2-trifluoroethyl)piperidin-4-yl)methyl alaninate dihydrochloride. The product (3.52 g) was prepared in a similar manner as described for Intermediate 13. 1 H NMR (400 MHz, DMSO-d6) δ 8.67 (s, 3H), 4.44 - 3.75 (m, 5H), 3.49 - 2.81 (m, 4H), 2.00 - 1.61 (m, 5H), 1.43 (d, J = 7.2 Hz, 3H). 19 F NMR (376 MHz, DMSO-d6) δ -63.30 (d, J = 443.2 Hz).

[0556]

[0557] (1-(2,2,2-trifluoroethyl)piperidin-4-yl)methyl ((4-nitrophenoxy)(phenoxy)phosphoryl)alaninate. The product (4.25 g) was prepared in a similar manner as described for Intermediate 35. 1 H NMR (400 MHz, DMSO-d6) δ 8.32 - 8.24 (m, 2H), 7.53 - 7.40 (m, 2H), 7.39 (ddd, J = 8.1, 6.8, 3.1 Hz, 2H), 7.24 (ddd, J = 17.4, 6.5, 1.6 Hz, 3H), 6.69 (ddd, J = 13.7, 10.0, 8.4 Hz, 1H), 4.07 - 3.92 (m, 1H), 3.88 - 3.77 (m, 2H), 3.08 (qd, J = 10.3, 1.6 Hz, 2H), 2.87 - 2.79 (m, 2H), 2.25 - 2.14 (m, 2H), 1.56 - 1.39 (m, 3H), 1.26 - 1.08 (m, 5H). 31 P NMR (162 MHz, DMSO-d6) δ -1.26, -1.49. 19F NMR (376 MHz, DMSO-d6) δ -68.45 (td, J = 10.2, 2.4 Hz). LCMS: MS m / z = 546.27 [M+l]; t R = 1.12 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Phenomenex Kinetex 2.6 μ XB-C18 100A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% acetonitrile, 1.8 min - 1.85 min 100% - 2% acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min.

[0558] Intermediate 39. ((4-nitrophenoxy)(pentoxy)phosphoryl)-L-alanine benzyl ester Intermediate 40. 4-nitrophenyl-N,N'-methyl L-alanine diaminophosphonate

[0559]

[0560] 4-((((BENZYLOXY)CARBONYL)-L-ALANINYL)OXY)-3,3-DIFLUOROPIPERIDINE-1- CARBOXYLIC ACID TERT-BUTYL ESTER. To a mixture of N-Cbz-L-alanine (2.0 g, 8.96 mmol), 3,3-difluoro-4-hydroxypiperidine-l-carboxylic acid tert-butyl ester (2.12 g, 8.96 mmol), and EDCI (1.67 g, 10.75 mmol) in acetonitrile (20 mL) was added DMAP (1.64 g, 13.44 mmol). The mixture was then stirred at room temperature for 15 hours, then diluted with EtOAc, washed with brine, dried over sodium sulfate, and concentrated in vacuo. The obtained residue was purified by silica gel chromatography (50% to 100% EtOAc in hexanes) to give the product. 19 F NMR (376 MHz, DMSO-d6) δ -68.45 (td, J = 10.2, 2.4 Hz). LCMS: MS m / z = 546.27 [M+l]; t R= 1.23 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Chromatography column: Phenomenex Kinetex 2.6μ XB-C18 100A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% acetonitrile, 1.8 min - 1.85 min 100% - 2% acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min.

[0561]

[0562] 3,3-Difluoropiperidin-4-yl ((benzyloxy)carbonyl)-L-alaninate. To a mixture of tert-butyl 4-((((benzyloxy)carbonyl)-L-alaninyl)oxy)-3,3-difluoropiperidine-1- carboxylate (330 mg, 0.746 mmol) in DCM (5 mL) was added slowly a 4 M solution of HCl in dioxane (0.9 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 hours, concentrated in vacuo, co-evaporated with DCM several times, and dried under high vacuum for 15 hours to give the product. 1 H NMR (400 MHz, Chloroform-d) δ 7.33 (m, 5H), 5.59 (m, 1H), 5.27 - 5.01 (m, 3H), 4.53 - 4.25 (m, 1H), 3.12 (m, 1H), 3.03 - 2.76 (m, 2H), 2.73 (s, 1H), 1.94 (s, 1H), 1.80 (s, 1H), 1.41 (d, J = 7.2 Hz, 3H). 19 F NMR (376 MHz, Chloroform-d) δ -114.66 (dd, J = 245.9, 61.8 Hz), -119.63.

[0563]

[0564] 1-Ethyl-3,3-difluoropiperidin-4-yl ((benzyloxy)carbonyl)-L-alaninate. A mixture of 3,3-difluoropiperidin-4-yl ((benzyloxy)carbonyl)-L-alaninate (450 mg, 1.190 mmol), acetaldehyde (0.194 mL, 2.629 mmol), and acetic acid (0.15 mL, 2.629 mmol) in DCM (9 mL) was stirred at room temperature for 20 min and sodium cyanoborohydride (330 mg, 5.258 mmol) was added. The resulting mixture was stirred for 1 hour and purified by preparative HPLC (Phenominex Gemini10u C18 30x100mm, 5u, 90A, 3mL / min, 20-60% ACN in water with 0.1% formic acid over 5 min, 60% ACN over 1 min, 60% ACN for 1 min, 2 min post time) to give the product. 250 x 21.2 mm column, 20-80% acetonitrile (0.1% TFA) / water (0.1% TFA) gradient) purification to give the product. 1 H NMR (400 MHz, Acetonitrile-d3) δ 10.18 (bs, 2H), 7.38 (m, 5H), 6.19 (m, 1H), 5.47 - 5.26 (m, 1H), 4.33 (m, 1H), 3.82 - 2.98 (m, 6H), 2.30 (s, 1H), 2.16 (s, 1H), 1.42 (m, 3H), 1.31 (td, J = 7.3, 1.5 Hz, 3H). LCMS: MS m / z = 371.27 [M+1]; t R = 0.66 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Phenomenex Kinetex 2.6 μ XB-C18 100A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% acetonitrile, 1.8 min - 1.85 min 100% - 2% acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min.

[0565]

[0566] 1 -Ethyl-3, 3-difluoropiperidin-4-yl L-alaninate. A mixture of 1 -ethyl-3, 3-difluoropiperidin-4-yl ((benzyloxy)carbonyl)-L-alaninate (450 mg, 0.929 mmol) and 20% Pd(OH)2 / C in THF (10 mL) was stirred under H2gas at room temperature for 1 hour, filtered, concentrated in vacuo, co-evaporated with DCM several times, and dried under high vacuum for 1 hour to give the product. LCMS: MS m / z = 237.09 [M+1]; t R = 0.66 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Phenomenex Kinetex 2.6 μ XB-C18 100A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% acetonitrile, 1.8 min - 1.85 min 100% - 2% acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min.

[0567]

[0568] (1 -Ethyl-3, 3-difluoropiperidin-4-yl ((4-nitrophenoxy)(phenoxy)phosphoryl)-L-alaninate. Methylene chloride (10 mL) was added to a slurry of 1 -ethyl-3, 3-difluoropiperidin-4-yl L-alaninate (480 mg, 1.37 mmol) and TEA (0.190 mL, 0.370 mmol) was added to obtain a solution which was cooled to -78 °C and phenyl dichlorophosphate (0.205 mL, 1.370 mmol) was added quickly. Triethylamine (0.190 mL, 1.37 mmol) was added over 30 min at -78 °C. The resulting mixture was stirred at the same temperature for 30 min and 4-nitrophenol (191 mg, 1.370 mmol) was added in one portion. Then triethylamine (0.190 mL, 1.370 mmol) was added over 30 min at -78 °C. The mixture was then stirred at room temperature for 2 hours, washed with water and brine, dried over sodium sulfate and concentrated in vacuo. The residue was then purified by silica gel column chromatography (0% to 100% EtOAc in hexanes) to give the product. 1 H NMR (400 MHz, Chloroform-d) δ 8.29 - 8.15 (m, 2H), 7.44 - 7.28 (m, 4H), 7.27 - 7.11 (m, 3H), 5.03 (m, 1H), 4.34 - 4.14 (m, 1H), 3.94 - 3.75 (m, 1H), 2.88 (s, 1H), 2.63-2.49 (m, 4H), 2.39 (m, 1H), 2.03 - 1.93 (m, 1H), 1.93 - 1.77 (m, 1H), 1.44 (m, 3H), 1.09 (td, J = 7.2, 1.0 Hz, 3H). 31 P NMR (162 MHz, Chloroform-d) δ -3.21, -3.26, -3.32, -3.46. 19 F NMR (377 MHz, Chloroform-d) δ -110.50 (d, J = 244.0 Hz), -116.76 (m). LCMS: MS m / z = 514.29 [M+l]; t R = 0.80 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Phenomenex Kinetex 2.6 μ XB-C18 100A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% acetonitrile, 1.8 min - 1.85 min 100% - 2% acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min.

[0569] Intermediate 41. ((4-nitrophenoxy)(pentoxy)phosphoryl)-L-alanine methyl ester

[0570]

[0571] To a solution of L-alanine ethyl ester HC1 salt (1.8 g, 11.72 mmol) in DCM (20 mL) was added 4-nitrobenzene phosphorodichloridate (1.5 g, 5.86 mmol) in one portion. The resulting mixture was cooled to 0 °C and triethylamine (2.37 g, 23.44 mmol) was added dropwise. After the ice bath was removed, the resulting mixture was stirred for 30 min and stirred overnight. The reaction mixture was then diluted with EtOAc, washed with water and brine, the organic solvent was concentrated in vacuo and the resulting residue was purified by silica gel column chromatography eluting with 0-100% ethyl acetate in hexanes to afford the product. LCMS: MS m / z = 417.93 [M+1], t R = 1.23 min; HPLC system: Agilent 1290 II; Column: Phenomenex Kinetex C18, 2.6u 110A, 100 x 4.6 mm; Solvents: A: Water with 0.1% TFA, B: Acetonitrile with 0.1% TFA; Gradient: 2% - 98% B, 8.5 min gradient, at a rate of 1.5 mL / min. R = 1.23 min; HPLC system: Agilent 1290 II; Column: Phenomenex Kinetex C18, 2.6u 110A, 100 x 4.6 mm; Solvents: A: Water with 0.1% TFA, B: Acetonitrile with 0.1% TFA; Gradient: 2% - 98% B, 8.5 min gradient, at a rate of 1.5 mL / min.

[0572] Intermediate 42. ((4-nitrophenoxy)(pentoxy)phosphoryl)-L-alanine methyl ester

[0573]

[0574] Phosphorous oxychloride (1.49 mL, 10 mmol) was dissolved in 20 mL of anhydrous dichloromethane and stirred in an ice bath under atmospheric nitrogen. L-alanine benzyl ester HC1 (2.2 g, 10 mmol) was added to the reaction solution in one portion and stirred for 10 min. Triethylamine (3 mL, 22 mmol) was dissolved in 5 mL of anhydrous dichloromethane and added dropwise to the reaction. The reaction mixture was stirred for 2 hours. p-Nitrophenol (1.25 g, 9 mmol) was added in one portion. Triethylamine (1.5 mL, 11 mmol) was dissolved in 3 mL of anhydrous dichloromethane and added dropwise to the reaction. The reaction mixture was stirred for 1 hour and diluted with dichloromethane (10 mL) and washed with water (3 x 10 mL). The organic extract was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude residue was purified via Si02column chromatography (4 g Si02Combiflash HP Gold column, 0% - 30% ethyl acetate / hexanes). Fractions containing the desired product were combined and concentrated under reduced pressure to yield the product. 1 H NMR (400 MHz, Chloroform-d) δ 8.24 - 8.10 (m, 2H), 7.40 - 7.10 (m, 12H), 5.14 (m, 2H), 4.19 (m, 1H), 3.87 (m, 1H), 1.47 - 1.36 (m, 3H). 31 P NMR (162 MHz, Chloroform-d) δ -3.15, -3.29. LCMS: MS m / z = 457.1 [M+1]; 455.1 [M-1], t R = 1.45 min; LC system: Thermo Dionex ultimate 3000 UHPLC; Column: Phenomenex Kinetex 2.6 μ C18 100 A, 50 x 3 mm; Solvents: A: water with 0.1% acetic acid, B: acetonitrile with 0.1% acetic acid; Gradient: 0 min - 0.3 min 5% B, 0.3 min - 1.5 min 5% - 100% B, 1.5 min - 2 min 100% B, 2 min - 2.2 min 100% - 5% B, at a rate of 2 mL / min. HPLC: t R = 4.03 min; HPLC system: Agilent 1100 series; Column: Phenomenex Gemini 5 μ C18 110 A, 50 x 4.6 mm; Solvents: A: water with 0.1% TFA, B: acetonitrile with 0.1% TFA; Gradient: 2% - 98% B over 5 min, at a rate of 2 mL / min.

[0575] Intermediate 43. ((4-(dimethylcarbamoyl)pentoxy)(4-nitrophenoxy)phosphoryl)-L-alanine isopropyl ester

[0576]

[0577] Triethylamine (3.68 mL, 26.4 mmol) was added to a solution of L-alanine methyl ester hydrochloride (1.63 g, 12.0 mmol) and 4-nitrophenyl phosphorodichloridate (1.5 g, 5.9 mmol) in dichloromethane (23 mL) at 0 °C under an argon atmosphere. After 3 h, the reaction mixture was diluted with dichloromethane (50 mL), washed with saturated aqueous sodium bicarbonate (50 mL) and brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude residue was purified by silica gel chromatography eluting with 0-100% ethyl acetate in hexanes to give the product. 1 H NMR (400 MHz, Chloroform-d1) 8.25 - 8.16 (m, 2H), 7.38 (dd, J = 9.3, 1.0 Hz, 2H), 4.17 - 3.95 (m, 2H), 3.73 (br s, 6H), 3.61 (br t, J = 10.0 Hz, 2H), 1.42 (s, 3H), 1.40 (s, 1H). 31 P NMR (162 MHz, Chloroform-d1) δ 7.82 (s). LCMS: MS m / z = 389.98 [M+l], t R = 1.11 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Kinetex 2.6 μ XB-C18 100A, 50 x 4.6 mm; Solvents: Acetonitrile with 0.1% acetic acid, Water with 0.1% acetic acid; Gradient: 0 min - 2.0 min 2% - 100% ACN, 2.0 min - 3.05 min 100% ACN, 3.05 min - 3.2 min 100% - 2% ACN, 3.2 min - 3.5 min 2% ACN, at a rate of 2 μL / min. HPLC: t R = 2.81 min; HPLC system: Agilent 1100 series; Column: Gemini 5 μ C18 110A, 50 x 4.6 mm; Solvents: Acetonitrile with 0.1% TFA, Water with 0.1% TFA; Gradient: 0 min - 5.0 min 2% - 98% ACN, 5.0 min - 6.0 min 98% ACN, at a rate of 2 mL / min.

[0578] Intermediate 44. oxetan-3-yl ((4-nitrophenoxy)(pentoxy)phosphoryl)-L-alanine ester

[0579]

[0580] Phosphorous oxychloride (2.81 mL, 18.9 mmol) and triethylamine (5.38 mL, 37.9 mmol) were added sequentially to a suspension of L-alanine methyl ester hydrochloride (2.64 g, 18.9 mmol) in dichloromethane (100 mL) at 0 °C. After 1 h, 4-nitrophenol (2.64 g, 18.9 mmol) and triethylamine (2.64 mL, 18.9 mmol) were added sequentially at 0 °C, and the resulting mixture was allowed to warm to RT. After 2.5 h, the reaction mixture was diluted with dichloromethane (100 mL), washed with saturated aqueous sodium bicarbonate (100 mL) and brine (100 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude residue was purified by silica gel chromatography eluting with 0-100% ethyl acetate in hexanes to give the product. 1 H NMR (400 MHz, Chloroform-d1) δ 8.25 - 8.18 (m, 2H), 7.43 - 7.29 (m, 4H), 7.29 - 7.15 (m, 3H), 4.24 - 4.07 (m, 1H), 3.97 (br q, J = 9.8 Hz, 1H), 3.70 (s, 3H), 1.45 - 1.35 (m, 3H). 31 PNMR (162 MHz, Chloroform-d1) δ -3.12 (s), -3.17 (s). LCMS: MS m / z = 380.98 [M+1], t R = 3.49 min; HPLC system: Agilent 1100 series; Column: Gemini 5μ C18 110A, 50 x 4.6 mm; Solvents: Acetonitrile with 0.1% TFA, Water with 0.1% TFA; Gradient: 0 min - 5.0 min 2% - 98% ACN, 5.0 min - 6.0 min 98% ACN, at a rate of 2 mL / min. R = 3.49 min; HPLC system: Agilent 1100 series; Column: Gemini 5μ C18 110A, 50 x 4.6 mm; Solvents: Acetonitrile with 0.1% TFA, Water with 0.1% TFA; Gradient: 0 min - 5.0 min 2% - 98% ACN, 5.0 min - 6.0 min 98% ACN, at a rate of 2 mL / min.

[0581] Intermediate 45. ((4-nitrophenoxy)(pentoxy)phosphoryl)-L-alanine propyl ester

[0582]

[0583] Under an argon atmosphere, 4-nitrobenzene phosphorodichloridate (2.00 g, 7.81 mmol) and triethylamine (2.18 mL, 15.6 mmol) were added sequentially to a suspension of L-alanine methyl ester hydrochloride (1.091 g, 18.9 mmol) in dichloromethane (23 mL) at 0 °C. After 1 h, benzyl alcohol (0.810 mL, 7.81 mmol) and triethylamine (1.09 mL, 7.81 mmol) were added sequentially at 0 °C and the resulting mixture was allowed to warm to RT. After 1 h, the reaction mixture was diluted with dichloromethane (50 mL), washed with saturated aqueous sodium bicarbonate (50 mL) and brine (50 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude residue was purified by silica gel chromatography eluting with 0-100% ethyl acetate in hexanes to give the product. 1 H NMR (400 MHz, Chloroform-d1) δ 8.32 - 8.09 (m, 2H), 8.32 - 8.09 (m, 7H), 5.15 (app t, J = 8.4 Hz, 2H), 4.70 (s, 1H), 4.07 - 3.93 (m, 1H), 3.73 - 3.65 (m, 3H), 1.42 - 1.31 (m, 3H). 31 PNMR (162 MHz, Chloroform-d1) δ 2.23 (s), 2.15 (s). LCMS: MS m / z = 394.9 [M+1], t R = 1.34 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Kinetex 2.6 μ XB-C18 100A, 50 x 4.6 mm; Solvents: Acetonitrile with 0.1% acetic acid, Water with 0.1% acetic acid; Gradient: 0 min - 2.0 min 2% - 100% acetonitrile, 2.0 min - 3.05 min 100% acetonitrile, 3.05 min - 3.2 min 100% - 2% acetonitrile, 3.2 min - 3.5 min 2% ACN, at a rate of 2 μL / min.

[0584] Intermediate 46. oxetan-3-ylmethyl ((4-nitrophenoxy)(pentoxy)phosphoryl)-L-alanine ester Intermediate 47. ((4-nitrophenoxy)(pentoxy)phosphoryl)-L-alanine cyclobutyl ester

[0585]

[0586] To a solution of 4-nitrobenzene phosphorodichloridate (620 mg, 2.422 mmol) and isopropyl L-alanine-HCl (406 mg, 2.422 mmol) in DCM-THF (10:3 mL) was added a solution of TEA (0.68 mL, 4.844 mmol) in DCM (3.32 mL) at -78 °C over 30 min. After removing the dry ice bath, the resulting mixture was stirred for 30 min and cooled to -78 °C, and N,N-dimethyl-4-hydroxybenzamide (400 mg, 2.422 mmol) was added in one portion, and a solution of TEA (0.34 mL, 2.422 mmol) in DCM (3.66 mL) was added at -78 °C over 30 min. After removing the dry ice bath, the resulting mixture was stirred for 1 hour, then diluted with EtOAc, washed with brine, concentrated in vacuo, and the resulting residue was purified by silica gel column chromatography (0% to 100% EtOAc in hexanes) to give the product. 1 H NMR (400 MHz, Chloroform-d) δ 8.26 - 8.18 (m, 2H), 7.45 - 7.35 (m, 3H), 7.27 (m, 2H), 6.76 (m, 1H), 5.01 (m, 1H), 4.17 - 3.94 (m, 2H), 3.19 - 2.84 (m, 6H), 1.39 (m, 3H), 1.27 - 1.16 (m, 6H). 31 P NMR (162 MHz, Chloroform-d) δ -3.13, -3.21. MS m / z = 480 (M+H). LCMS: MS m / z = 480.26 [M+1]; t R = 1.00 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Phenomenex Kinetex 2.6 μ XB-C18 100A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% acetonitrile, 1.8 min - 1.85 min 100% - 2% acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min.

[0587] Intermediate 50. ((S)-(perfluorophenoxy)(pentoxy)phosphoryl)-L-alanine methyl ester

[0588]

[0589] Oxetan-3-yl((benzyloxy)carbonyl)-L-alaninate. To a mixture of ((benzyloxy)carbonyl)-L-alanine (1.8 g, 8.1 mmol), 3-hydroxyoxetane (0.5 g, 6.75 mmol) and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide HC1 salt (EDCI) (1.68 g, 8.77 mmol) in acetonitrile (100 mL) was added 4-(dimethylamino)pyridine (DMAP, 1.24 g, 10.12 mmol). The mixture was then stirred at room temperature for 2 hours, then the reaction mixture was diluted with EtOAc, washed with brine, the organic solvent was dried over sodium sulfate, then concentrated in vacuo. The obtained residue was purified by silica gel chromatography eluting with 0-100% ethyl acetate in hexanes to give the product. 1 H NMR (400 MHz, Chloroform-d) δ 7.40 - 7.28 (m, 5H), 5.47 (p, J = 5.9 Hz, 1H), 5.30 (d, J = 8.0 Hz, 1H), 5.10 (s, 2H), 4.88 (t, J = 7.1 Hz, 2H), 4.62 (ddd, J = 17.5, 7.7, 5.3 Hz, 2H), 4.41 (p, J = 7.3 Hz, 1H), 1.44 (d, J = 7.3 Hz, 3H). LCMS: MS m / z = 280.04 [M+l], t R = 1.11 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Kinetex 2.6 u XB-C18 100 A, 50 x 4.6 mm; Solvents: Acetonitrile with 0.1% acetic acid, water with 0.1% acetic acid; Gradient: 0 min - 2.0 min 2% - 100% acetonitrile, 2.0 min - 3.05 min 100% acetonitrile, 3.05 min - 3.2 min 100% - 2% acetonitrile, 3.2 min - 3.5 min 2% ACN, at a rate of 2 uL / min. HPLC: t R = 2.82 min; HPLC system: Agilent 1290 II; Column: Phenomenex Kinetex C18, 2.6 u 10 A, 100 x 4.6 mm; Solvents: A: water with 0.1% TFA, B: acetonitrile with 0.1% TFA; Gradient: 2% - 98% B, 8.5 min gradient, at a rate of 1.5 mL / min.

[0590]

[0591] Oxetan-3-yl L-alaninate. Oxetan-3-yl ((benzyloxy)carbonyl)-L-alaninate (0.1 g, 0.36 mmol) was dissolved in DCM (5 mL) to this solution was added 15 mg Pd-C (10%, wet), the reaction flask was degassed and then charged with a H2 balloon, stirred at RT for 2 h, then the reaction mixture was filtered, the solvent was evaporated under vacuum, the residue was dried under high vacuum for 5 min to get the product. 1 H NMR (400 MHz, Chloroform-d) δ 5.42 (p, J = 5.7 Hz, 1H), 4.87 (t, J = 6.9 Hz, 2H), 4.65 - 4.54 (m, 2H), 3.58 (qd, J = 7.0, 2.1 Hz, 1H), 1.49 (d, J = 7.1 Hz, 2H), 1.34 (dd, J = 7.2, 2.1 Hz, 3H).

[0592]

[0593] Oxetan-3-yl ((4-nitrophenoxy)(phenoxy)phosphoryl)-L-alaninate. To a solution of oxetan-3-yl L-alaninate (120 mg, 0.83 mmol) in DCM (10 mL) was added phenyl dichlorophosphate (175 mg, 0.83 mmol) in one portion. The resulting mixture was cooled to 0 °C and triethylamine (252 mg, 2.49 mmol) was added dropwise. After removing the ice bath, the resulting mixture was stirred for 30 min and cooled to 0 °C and p-nitrophenol (115 mg, 0.83 mmol) was added in one portion, triethylamine (252 mg, 2.49 mmol) was added dropwise. After removing the ice bath, the resulting mixture was stirred for 30 min, diluted with EtOAc, washed with water and brine, the organic solvent was concentrated under vacuum and the resulting residue was purified by silica gel column chromatography eluting with 0% - 100% ethyl acetate in hexanes to get the product. LCMS: MS m / z = 423.06 [M+1], t R = 1.25 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Kinetex 2.6 μ XB-C18 100A, 50 x 4.6 mm; Solvents: Acetonitrile with 0.1% acetic acid, water with 0.1% acetic acid; Gradient: 0 min - 2.0 min 2% - 100% acetonitrile, 2.0 min - 3.05 min 100% acetonitrile, 3.05 min - 3.2 min 100% - 2% acetonitrile, 3.2 min - 3.5 min 2% ACN, at a rate of 2 μL / min. HPLC: t R= 3.15 min; HPLC system: Agilent 1290 II; Column: Phenomenex Kinetex C18, 2.6u 110A, 100 x 4.6 mm; Solvents: A: water with 0.1% TFA, B: acetonitrile with 0.1% TFA; Gradient: 2% - 98% B over 8.5 min gradient at a rate of 1.5 mL / min.

[0594] Intermediate 51. ((4-(2-methoxyethoxy)pentoxy)(4-nitrophenoxy)phosphoryl)-L-alanine isopropyl ester

[0595]

[0596] (tert-Butoxycarbonyl)-L-alanine propyl ester. At RT, N-(3-dimethylaminopropyl)-N'- ethylcarbodiimide hydrochloride (6.08 g, 31.71 mmol) was added to a solution of Boc-Ala-OH (5 g, 26.43 mmol) and n-propanol (6.02 mL, 80.6 mmol) in acetonitrile (125 mL). After 15 min, 4-(dimethylamino)pyridine (3.23 g, 26.43 mmol) was added. After 16 h, the reaction mixture was concentrated to half volume, the mixture was diluted with ethyl acetate (250 mL), and the resulting mixture was washed with saturated aqueous sodium carbonate solution (2 x 200 mL) and brine (200 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude residue was subjected to silica gel chromatography eluting with 0% - 20% EtOAc in hexanes to give the product. 1 H NMR (400 MHz, Acetonitrile-d3) δ 5.57 (s, 1H), 4.19 - 3.92 (m, 3H), 1.63 (h, J = 7.1 Hz, 2H), 1.40 (s, 9H), 1.30 (d, J = 7.3 Hz, 3H), 0.93 (t, J = 7.4 Hz, 3H). LCMS: MS m / z = 231.60 [M+l], t R = 1.10 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Kinetex 2.6 μ XB-C18 100A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% acetonitrile, 1.8 min - 1.85 min 100% - 2% acetonitrile, 1.85 min - 2.00 min 2% ACN at a rate of 1800 μL / min.

[0597]

[0598] L-alanine propyl ester hydrochloride. At RT, a 4 M solution of hydrochloric acid in dioxane (16.91 mL) was added to a solution of (tert-butoxycarbonyl)-L-alanine propyl ester (3.91 g, 16.91 mmol) in dichloromethane (10 mL). After 16 h, the reaction mixture was concentrated under reduced pressure to give the product. 1 H NMR (400 MHz, Acetonitrile-d3) δ 8.45 (s, 3H), 4.22 - 4.11 (m, 2H), 4.11 - 3.99 (m, 1H), 1.68 (dtd, J = 14.0, 7.4, 6.6 Hz, 2H), 1.60 (d, J = 7.2 Hz, 3H), 0.95 (t, J = 7.4 Hz, 3H). LCMS: MS m / z = 131.94 [M+1], t R = 0.32 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Kinetex 2.6 μ XB-C18 100A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% acetonitrile, 1.8 min - 1.85 min 100% - 2% acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min.

[0599]

[0600] ((4-nitrophenoxy)(phenoxy)phosphoryl)-L-alanine propyl ester. At 0 °C, a solution of phenyl dichlorophosphate (0.89 mL, 5.97 mmol) in dichloromethane (12 mL) was added dropwise to a solution of L-alanine propyl ester hydrochloride (1.0 g, 5.97 mmol) in dichloromethane (12 mL) over 15 min. After the addition was complete, a solution of triethylamine (2.0 mL, 14.32 mmol) in dichloromethane (2.5 mL) was added over 5 min. After 3.5 h, 4-nitrophenol (0.83 g, 5.97 mmol) and triethylamine (1.0 mL, 7.16 mmol) were added sequentially at 0 °C, and the resulting mixture was allowed to warm to RT. After 2 h, the reaction mixture was diluted with dichloromethane (50 mL), washed with water (2 x 100 mL) and brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude residue was purified by silica gel chromatography eluting with 0% - 100% ethyl acetate in hexanes to give the product. 1H NMR (400 MHz, Acetonitrile-d3) δ 8.28 - 8.20 (m, 2H), 7.49 - 7.35 (m, 4H), 7.31 - 7.19 (m, 3H), 4.72 - 4.56 (m, 1H), 4.14 - 4.02 (m, 1H), 3.99 (td, J = 6.6, 2.5 Hz, 2H), 1.58 (dtdd, J = 13.9, 7.4, 6.5, 0.9 Hz, 2H), 1.31 (ddd, J = 7.1, 4.2, 1.1 Hz, 3H), 0.88 (t, J = 7.4 Hz, 3H). 31 P NMR (162 MHz, Acetonitrile-d3) δ -2.12, -2.22. LCMS: MS m / z = 409.12 [M+1], t R = 1.15 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Kinetex 2.6 μ XB-C18 100 A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% acetonitrile, 1.8 min - 1.85 min 100% - 2% acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min. HPLC: t R = 5.73 min; HPLC system: Agilent 1100 series; Column: Gemini 5 μ C18 110 A, 50 x 4.6 mm; Solvents: Acetonitrile with 0.1% TFA, water with 0.1% TFA; Gradient: 0 min - 9.0 min 2% - 95% ACN, 9.0 min - 10.0 min 95% ACN, at a rate of 2 mL / min.

[0601] Intermediate 52. ((4-nitrophenoxy)(pentoxy)phosphoryl)-L-alanine butyl ester

[0602]

[0603] Oxetan-3-ylmethyl ((benzyloxy)carbonyl)-L-alaninate. To a mixture of ((benzyloxy)carbonyl)-L-alanine (6.08 g, 27.24 mmol), oxetan-3-ylmethanol (2 g, 22.7 mmol) and l-ethyl-3-(3-dimethylaminopropyl)carbodiimide HC1 salt (EDCI) (5.66 g, 29.51 mmol) in acetonitrile (100 mL) was added 4-(dimethylamino)pyridine (DMAP, 4.16 g, 34.05 mmol). The mixture was then stirred at room temperature for 2 hours, the reaction mixture was then diluted with EtOAc, washed with brine, the organic solvent was dried over sodium sulfate and then concentrated in vacuo. The obtained residue was purified by silica gel chromatography eluting with 0-100% ethyl acetate in hexane to obtain the product. LCMS: MS m / z = 280.04 [M+1], t R = 1.11 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Kinetex 2.6 μ XB-C18 100A, 50 x 4.6 mm; Solvents: Acetonitrile with 0.1% acetic acid, water with 0.1% acetic acid; Gradient: 0 min - 2.0 min 2% - 100% acetonitrile, 2.0 min - 3.05 min 100% acetonitrile, 3.05 min - 3.2 min 100% - 2% acetonitrile, 3.2 min - 3.5 min 2% ACN, at a rate of 2 μL / min. HPLC: t R = 2.88 min; HPLC system: Agilent 1290 II; Column: Phenomenex Kinetex C18, 2.6u 110A, 100 x 4.6 mm; Solvents: A: water with 0.1% TFA, B: acetonitrile with 0.1% TFA; Gradient: 2% - 98% B, 8.5 min gradient, at a rate of 1.5 mL / min.

[0604]

[0605] Oxetan-3-ylmethyl L-alaninate. Oxetan-3-ylmethyl ((benzyloxy)carbonyl)-L-alanine ester (2.2 g, 8 mmol) was dissolved in DCM (25 mL), to this solution was added 500 mg Pd-C (10%, wet), the reaction flask was degassed and then charged with a H2 balloon, stirred at RT for 2 hours, the reaction mixture was then filtered, the solvent was evaporated under vacuum, the residue was dried under high vacuum for 5 min to obtain the product. 1H NMR (400 MHz, Chloroform-d) δ 4.77 (dd, J = 7.9, 6.3 Hz, 2H), 4.44 (td, J = 6.1, 2.5 Hz, 2H), 4.38 - 4.23 (m, 2H), 3.55 (q, J = 7.0 Hz, 1H), 3.34 - 3.19 (m, 1H), 1.31 (d, J = 7.0 Hz, 3H).

[0606]

[0607] Oxetan-3-ylmethyl ((4-nitrophenoxy)(phenoxy) phosphoryl)-L-alaninate. To a solution of oxetan-3-ylmethyl L-alaninate (1.19 g, 7.11 mmol) in DCM (20 mL) was added phenyl dichlorophosphate (1.5 g, 7.11 mmol) in one portion. The resulting mixture was cooled to 0 °C and triethylamine (1.44 g, 14.22 mmol) was added dropwise. After removing the ice bath, the resulting mixture was stirred for 30 min and cooled to 0 °C, and p-nitrophenol (0.99 g, 7.1 mmol) was added in one portion, and triethylamine (1.44 g, 14.22 mmol) was added dropwise. After removing the ice bath, the resulting mixture was stirred for 30 min, diluted with EtOAc, washed with water and brine, the organic solvent was concentrated in vacuo, and the resulting residue was purified by silica gel column chromatography eluting with 0% - 100% ethyl acetate in hexanes to give the product. LCMS: MS m / z = 437.14 [M+1], t R = 1.25 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Kinetex 2.6 u XB-C18 100A, 50 x 4.6 mm; Solvents: Acetonitrile with 0.1% Acetic acid, Water with 0.1% Acetic acid; Gradient: 0 min - 2.0 min 2% - 100% Acetonitrile, 2.0 min - 3.05 min 100% Acetonitrile, 3.05 min - 3.2 min 100% - 2% Acetonitrile, 3.2 min - 3.5 min 2% ACN, at a rate of 2 uL / min. HPLC: t R = 3.36 min; HPLC system: Agilent 1290 II; Column: Phenomenex Kinetex C18, 2.6u 110A, 100 x 4.6 mm; Solvents: A: Water with 0.1% TFA, B: Acetonitrile with 0.1% TFA; Gradient: 2% - 98% B, 8.5 min gradient, at a rate of 1.5 mL / min.

[0608] Intermediate 53. ((4-nitrophenoxy)(pentoxy)phosphoryl)-L-alanine 3-methoxypropyl ester

[0609]

[0610] To a solution of L-alanine cyclobutyl ester (1.8 g, 10 mmol) in DCM (10 mL) was added phenyl phosphorodichloridate (2.1 g, 10 mmol) in one portion under nitrogen atmosphere in an ice bath. Then triethylamine (1.11 g, 11 mmol) was added dropwise. After removing the ice bath, the resulting mixture was stirred for 2 hours and cooled to 0 °C, and p-nitrophenol (2.5 g, 18 mmol) was added in one portion, and triethylamine (1.11 g, 11 mmol) was added dropwise. After removing the ice bath, the resulting mixture was stirred for 2 hours, diluted with EtOAc, washed with 5% aqueous citric acid solution twice, followed by brine, the organic solvent was concentrated in vacuo, and the resulting residue was purified by silica gel column chromatography eluted with 0-100% ethyl acetate in hexanes to give the product. MS m / z = 422.0 (M+H) + .

[0611] Resolution of Sp and Rp diastereomers. The product was purified via chiral preparative HPLC (Chiralpak IA, 150 x 4.6 mm, heptane 70%, IPA 30%) to form intermediate 48 and intermediate 49:

[0612]

[0613] Intermediate 48. First eluting diastereomer of intermediate 47: 1 H NMR (400 MHz, Methanol-d4) δ 8.33 - 8.23 (m, 2H), 7.52 - 7.33 (m, 4H), 7.33 - 7.17 (m, 3H), 4.96 - 4.85 (m, 1H), 4.07 - 3.96 (m, 1H), 2.27 (m, 2H), 2.07 - 1.91 (m, 2H), 1.83 - 1.70 (m, 1H), 1.70 - 1.55 (m, 1H), 1.32 (ddd, J=7.2, 5.3, 1.2 Hz, 3H). 31 P NMR (162 MHz, Methanol-d4) δ 1.36. LCMS: MS m / z = 421.05 [M+1], t R= 1.42 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Kinetex 2.6 μ XB-C18 100A, 50 x 4.6 mm; Solvents: Acetonitrile with 0.1% acetic acid, water with 0.1% acetic acid; Gradient: 0 min - 2.0 min 2% - 100% acetonitrile, 2.0 min - 3.05 min 100% acetonitrile, 3.05 min - 3.2 min 100% - 2% acetonitrile, 3.2 min - 3.5 min 2% ACN, at a rate of 2 μL / min. HPLC: t R = 8.07 min; HPLC system: Chiralpak IC, 150 x 4.6 mm, 5 micron, CN = IC00CD-QC005, 1 CV = 2.49 mL, CV#1, Column Valve: Position 3, 15 mL / 15 min, at a rate of 1 mL / min. Pmax = 300 bar; Solvent Valve: D: Heptane 70%, #6: IPA.

[0614] Intermediate 49. Second eluting diastereomer of Intermediate 47: 1 H NMR (400 MHz, Methanol-d4) δ 8.33 - 8.23 (m, 2H), 7.52 - 7.33 (m, 4H), 7.33 - 7.17 (m, 3H), 4.96 - 4.85 (m, 1H), 4.07 - 3.96 (m, 1H), 2.27 (m, 2H), 2.07 - 1.91 (m, 2H), 1.83 - 1.70 (m, 1H), 1.70 - 1.55 (m, 1H), 1.32 (ddd, J = 7.2, 5.3, 1.2 Hz, 3H). 31 P NMR (162 MHz, Methanol-d4) δ 1.59. LCMS: MS m / z = 420.90 [M+1], t R = 1.42 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Kinetex 2.6 μ XB-C18 100A, 50 x 4.6 mm; Solvents: Acetonitrile with 0.1% acetic acid, water with 0.1% acetic acid; Gradient: 0 min - 2.0 min 2% - 100% acetonitrile, 2.0 min - 3.05 min 100% acetonitrile, 3.05 min - 3.2 min 100% - 2% acetonitrile, 3.2 min - 3.5 min 2% ACN, at a rate of 2 μL / min. HPLC: t R= 11.50 min; HPLC system: Chiralpak IC, 150 x 4.6 mm, 5 microns, CN = IC00CD-QC005, 1 CV = 2.49 mL, CV#1, column valve: position 3, 15 mL / 15 min, rate 1 mL / min. Pmax = 300 bar; Solvent valve: D: Heptane 70%, #6: IPA 30%.

[0615] Intermediate 54. methyl (2S)-2-(((benzyloxy)carbonyl)amino)-3-(4-(((((S)-l-methoxy-l- oxopropan-2-yl)amino)(4-nitrophenoxy)phosphoryl)oxy)phenyl)propanoate

[0616]

[0617] L-alanine methyl ester hydrochloride (14 g, 100 mmol) was mixed with 50 mL of dry DCM and stirred in an ice bath under atmospheric nitrogen. Phenyl dichlorophosphate (16.4 mL, 110 mmol) was added dropwise to the reaction and the reaction mixture was stirred for 30 min. Triethylamine (29.4 mL, 210 mmol) was mixed with 20 mL of dry DCM and added dropwise to the reaction. The reaction was stirred for 1 hour. Pentafluorophenol (18.4 g, 100 mmol) was added in one portion. Triethylamine (14.7 mL, 105 mmol) was mixed with 30 mL of dry DCM and added dropwise to the reaction. The reaction mixture was stirred at RT for 16 hours.

[0618] The reaction was diluted with DCM (50 mL) and washed with water (5 x 10 mL). The organics were dried over anhydrous sodium sulfate and then concentrated under reduced pressure to give a solid. To the solid was added isopropyl ether (130 mL). The large solid chunks were broken up and then sonicated for 20 min before the mixture was stirred for 24 hours.

[0619] The solid was collected and washed with a small amount of isopropyl ether (30 mL). The solid was dried under high vacuum to give the product. 1 H NMR (400 MHz, Chloroform-d) δ 7.40 - 7.32 (m, 2H), 7.28 - 7.19 (m, 3H), 4.20 (m, 1H), 3.96 - 3.85 (m, 1H), 3.74 (s, 3H), 1.47 (d, J = 7.1 Hz, 3H). 31 P NMR (162 MHz, Chloroform-d) δ -1.62. 19 F NMR (376 MHz, Chloroform-d) δ -153.82 (dd, J = 18.5, 2.7 Hz), -159.99 (td, J = 21.8, 3.8 Hz), -162.65 (dd, J = 22.2, 17.6 Hz). LCMS: MS m / z = 425.9 [M+1], 423.9 [M-1], t R= 1.68 min; LC system: Thermo Dionex Ultimate 3000 UHPLC; column: Phenomenex Kinetex 2.6 μ C18 100A, 50 x 3 mm; solvents: A: water with 0.1 % acetic acid, B: acetonitrile with 0.1 % acetic acid; gradient: 0 min - 0.3 min 5 % B, 0.3 min - 1.5 min 5 % - 100 % B, 1.5 min - 2 min 100 % B, 2 min - 2.2 min 100 % - 5 % B, at a rate of 2 mL / min. HPLC: t R = 3.76 min; HPLC system: Agilent 1100 series; column: Phenomenex Gemini 5 μ C18 110A, 50 x 4.6 mm; solvents: A: water with 0.1 % TFA, B: acetonitrile with 0.1 % TFA; gradient: 2 % - 98 % B in 5 min, at a rate of 2 mL / min.

[0620] Intermediate 55. (S)-tetrahydrofuran-3-yl ((4-nitrophenoxy)(pentoxy)phosphoryl)-L-alanine ester Intermediate 56.

[0621]

[0622] A solution of 4-nitrobenzene phosphorodichloridate (503 mg, 1.97 mmol) in dichloromethane (20 mL) was added dropwise to a solution of L-alanine isopropyl ester hydrochloride (329 mg, 1.97 mmol) in dichloromethane (20 mL) at 0 °C over 10 min. After the addition was complete, triethylamine (0.55 mL, 3.93 mmol) was added dropwise. After 90 min, 4-(2-methoxy-ethoxy)phenol (331 mg, 1.97 mmol) and triethylamine (0.28 mL, 1.97 mmol) were added sequentially at 0 °C and the resulting mixture was allowed to warm to RT. After 30 min, the reaction mixture was washed with water (2 x 50 mL) and brine (50 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude residue was purified by silica gel chromatography eluting with 20 % - 100 % ethyl acetate in hexanes to give the product. 1H NMR (400 MHz, Methanol-d4) δ 8.32 - 8.24 (m, 2H), 7.51 - 7.39 (m, 2H), 7.24 - 7.12 (m, 2H), 6.97 - 6.90 (m, 2H), 4.94 (heptd, J = 6.2, 3.2 Hz, 1H), 4.12 - 4.07 (m, 2H), 4.05 - 3.93 (m, 1H), 3.76 - 3.68 (m, 2H), 3.41 (d, J = 0.5 Hz, 3H), 1.32 (td, J = 7.1, 1.2 Hz, 3H), 1.19 (dt, J = 6.3, 2.0 Hz, 6H). 31 PNMR (162 MHz, Methanol-d4) δ -0.86, -1.06. LCMS: MS m / z = 483.06 [M+l], t R = 1.39 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Kinetex 2.6 μ XB-C18 100A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% acetonitrile, 1.8 min - 1.85 min 100% - 2% acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min. HPLC: t R = 5.58 min; HPLC system: Agilent 1100 series; Column: Gemini 5 μ C18 110A, 50 x 4.6 mm; Solvents: Acetonitrile with 0.1% TFA, water with 0.1% TFA; Gradient: 0 min - 9.0 min 2% - 95% ACN, 9.0 min - 10.0 min 95% ACN, at a rate of 2 mL / min.

[0623] 3-morpholinopropyl ((4-nitrophenoxy)(pentoxy)phosphoryl)-L-alanine ester

[0624]

[0625] A solution of phenyl phosphorodichloridate (0.89 mL, 5.97 mmol) in dichloromethane (12 mL) was added dropwise to a solution of L-alanine butyl ester hydrochloride (CAS # 81305-85-3, 1.0 g, 5.97 mmol) in dichloromethane (12 mL) at 0 °C over 15 min. After the addition was complete, a solution of triethylamine (2.0 mL, 14.32 mmol) in dichloromethane (2.5 mL) was added over 5 min. After 3.5 h, 4-nitrophenol (0.83 g, 5.97 mmol) and triethylamine (1.0 mL, 7.16 mmol) were added sequentially at 0 °C and the resulting mixture was allowed to warm to RT. After 2 h, the reaction mixture was diluted with dichloromethane (50 mL), washed with water (2 x 100 mL) and brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude residue was purified by silica gel chromatography eluting with 0-100% ethyl acetate in hexanes to give the product. 1 H NMR (400 MHz, Methanol-d4) δ 8.31 - 8.23 (m, 1H), 7.52 - 7.34 (m, 2H), 7.32 - 7.18 (m, 2H), 4.04 (td, J = 6.6, 2.7 Hz, 2H), 1.60 - 1.48 (m, 1H), 1.40 - 1.26 (m, 3H), 0.89 (t, J = 7.4 Hz, 2H). 31 P NMR (162 MHz, Methanol-d4) δ -1.36, -1.59. LCMS: MS m / z = 423.13 [M+l], t R = 1.22 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Kinetex 2.6 μ XB-C18 100A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, Water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% acetonitrile, 1.8 min - 1.85 min 100% - 2% acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min.

[0626] Intermediate 57. (R)-tetrahydrofuran-3-yl ((4-nitrophenoxy)(pentoxy)phosphoryl)-L-alanine ester

[0627]

[0628] L-alanine 3-methoxypropyl ester. To a mixture of Cbz-L-alanine (2.80 g, 12.54 mmol), 3-methoxypropanol (1.00 mL, 10.45 mmol), and EDCI (2.11 g, 13.59 mmol) in acetonitrile (40 mL) was added DMAP (1.92 g, 15.68 mmol). The mixture was then stirred at room temperature for 15 hours, then diluted with EtOAc, washed with brine, dried over sodium sulfate, and concentrated in vacuo. The obtained residue was purified by silica gel chromatography (0% to 50% EtOAc in hexanes, 35 min run) to give Cbz-L-alanine ester (2.78 g), which was dissolved in THF (20 mL) and 20% Pd(OH)2(800 mg, 1.14 mmol) was added at room temperature. The resulting mixture was stirred under a hydrogen atmosphere at room temperature for 4 hours, filtered, concentrated in vacuo, and dried under high vacuum to give the product. 1 H NMR (400 MHz, Chloroform-d) δ 4.28 - 4.14 (m, 2H), 3.55 (q, J = 7.0 Hz, 1H), 3.43 (t, J = 6.2 Hz, 2H), 3.32 (s, 3H), 1.98 - 1.85 (m, 4H), 1.33 (d, J = 7.0 Hz, 3H). LCMS m / z = 161.98 (M+H), t R = 0.12 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Chromatography column: Phenomenex Kinetex 2.6 μ XB-C18 100A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% acetonitrile, 1.8 min - 1.85 min 100% - 2% acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min.

[0629]

[0630] ((4-nitrophenoxy)(phenoxy)phosphoryl)-L-alanine 3-methoxypropyl ester. To a solution of L-alanine 3-methoxypropyl ester (1.32 g, 8.20 mmol) in DCM (20 mL) at -78 °C was added phenyl dichlorophosphate (1.23 mL, 8.20 mmol) in one portion rapidly. Then triethylamine (1.14 mL, 8.20 mmol) was added at -78 °C within 5 min. After removing the dry ice bath, the resulting mixture was stirred for 30 min and cooled to -78 °C. p-Nitrophenol (1.14 g, 8.20 mmol) was added in one portion and triethylamine (1.14 mL, 8.20 mmol) was added at -78 °C within 5 min. After removing the dry ice bath, the resulting mixture was stirred for 2 hours. After dilution with DCM, the mixture was washed with brine, concentrated in vacuo, and the resulting residue was purified by silica gel column chromatography (0% to 100% EtOAc in hexanes) to give the product. 1 H NMR (400 MHz, Chloroform-d) δ 8.26 - 8.19 (m, 2H), 7.36 (m, 4H), 7.27 - 7.15 (m, 3H), 4.20 (m, 2H), 4.17 - 4.06 (m, 1H), 3.91 (m, 1H), 3.40 (m, 2H), 3.30 (m, 3H), 1.87 (m, 2H), 1.40 (m, 3H). 31 P NMR (162 MHz, Chloroform-d) δ -3.07, -3.10. LCMS: m / z = 439.11 (M+H), t R = 1.36 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Phenomenex Kinetex 2.6 μ XB-C18 100A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% acetonitrile, 1.8 min - 1.85 min 100% - 2% acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min

[0631] Intermediate 58. (chloro(pentoxy)phosphorothioyl)-L-alanine methyl ester Intermediate 59. ((((S)-l-(2-ethylbutoxy)-l-oxopropan-2-yl)amino)(4-nitrophenoxy)phosphoryl)-L-alanine methyl ester

[0632]

[0633] A solution of L-alanine methyl ester hydrochloride (275 mg, 1.97 mmol) in dichloromethane (20 mL) was added dropwise to a solution of 4-nitrophenyl phosphorodichloridate (504 mg, 1.97 mmol) in dichloromethane (20 mL) at 0 °C over 10 min. After the addition was complete, triethylamine (0.55 mL, 3.93 mmol) was added dropwise. After 60 min, N- benzyloxycarbonyl-L-tyrosine methyl ester (649 mg, 1.97 mmol) and triethylamine (0.28 mL, 1.97 mmol) were added sequentially at 0 °C, then the resulting mixture was allowed to warm to RT. After 3 h, the reaction mixture was washed with water (2 x 50 mL) and brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude residue was purified by silica gel chromatography eluting with 0-100% ethyl acetate in hexanes to give the product. 1 H NMR (400 MHz, Methanol-d4) δ 8.34 - 8.17 (m, 2H), 7.53 - 7.37 (m, 2H), 7.37 - 7.09 (m, 9H), 5.02 (s, 2H), 4.43 (dd, J = 9.4, 5.2 Hz, 1H), 4.19 - 3.97 (m, 1H), 3.70 (s, 3H), 3.62 (d, J = 4.4 Hz, 3H), 3.16 (dd, J = 14.0, 5.4 Hz, 1H), 2.93 (dd, J = 14.1, 9.8 Hz, 1H), 1.32 (td, J = 7.3, 1.2 Hz, 3H). 31 PNMR (162 MHz, Methanol-d4) δ -1.30, -1.51. LCMS: MS m / z = 616.03 [M+1], t R = 1.63 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Kinetex 2.6 μ XB-C18 100 A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% acetonitrile, 1.8 min - 1.85 min 100% - 2% acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min. HPLC: t R = 5.81 min; HPLC system: Agilent 1100 series; Column: Gemini 5 μ C18 110 A, 50 x 4.6 mm; Solvents: Acetonitrile with 0.1% TFA, water with 0.1% TFA; Gradient: 0 min - 9.0 min 2% - 95% ACN, 9.0 min - 10.0 min 95% ACN, at a rate of 2 mL / min.

[0634] ​

[0635]

[0636] (S)-tetrahydrofuran-3-yl-L-alanine ester. To a mixture of N-Cbz-L-alanine (3.31, 14.83 mmol), (S)-THF-3-ol (1.0 mL, 12.34 mmol), and EDCI (2.49 g, 16.04 mmol) in acetonitrile (20 mL) was added DMAP (2.26 g, 18.51 mmol). The mixture was then stirred at room temperature for 15 hours, then diluted with EtOAc, washed with brine, dried over sodium sulfate, and concentrated in vacuo. The obtained residue was purified by silica gel chromatography (0% to 80% EtOAc in hexanes) to give Cbz-L-alanine 4-THF ester, which was dissolved in THF (20 mL) and 20% palladium hydroxide (433 mg, 0.617 mmol) was added at room temperature. The resulting mixture was stirred under H2gas at room temperature for 2 hours, filtered, and concentrated in vacuo, co-evaporated with DCM multiple times, and dried under high vacuum for 15 hours to give the product. 1 H NMR (400 MHz, Chloroform-d) δ 5.37 - 5.29 (m, 1H), 3.97 - 3.77 (m, 4H), 3.61 - 3.52 (m, 1H), 2.27 - 2.12 (m, 1H), 2.02 (dt, J = 12.8, 5.6 Hz, 1H), 1.76 (s, 2H), 1.34 (dd, J = 7.1, 1.5 Hz, 3H). LCMS m / z = 159.94 (M+H), t R = 0.12 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Chromatography column: Phenomenex Kinetex 2.6 μ XB-C18 100A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% acetonitrile, 1.8 min - 1.85 min 100% - 2% acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min.

[0637]

[0638] (S)-tetrahydrofuran-3-yl ((4-nitrophenoxy)(phenoxy)phosphoryl)-L-alaninate. To a solution of (S)-tetrahydrofuran-3-yl-L-alaninate (1.45 g, 9.10 mmol) in DCM (20 mL) at -78 °C was added phenyl dichlorophosphate (1.37 mL, 9.10 mmol) in one portion rapidly. Then triethylamine (1.27 mL, 9.10 mmol) was added at -78 °C within 5 min. After removing the dry ice bath, the resulting mixture was stirred for 30 min and cooled to -78 °C. p-Nitrophenol (1.27 g, 9.10 mmol) was added in one portion and triethylamine (1.27 mL, 9.10 mmol) was added at -78 °C within 5 min. After removing the dry ice bath, the resulting mixture was stirred for 2 hours. After dilution with DCM, the mixture was washed with brine, concentrated in vacuo and the resulting residue was purified by silica gel column chromatography (0% to 100% EtOAc in hexanes) to give the product. 1 HNMR (400 MHz, Chloroform-d) δ 8.22 (m, 2H), 7.49 - 7.31 (m, 4H), 7.30 - 7.12 (m, 3H), 5.29 (m, 1H), 4.14 (m, 1H), 4.00 - 3.79 (m, 4H), 3.82 - 3.60 (m, 1H), 2.17 (m, 1H), 1.95 (m, 1H), 1.40 (m, 3H). 31 P NMR (162 MHz, Chloroform-d) δ -3.18, -3.20. LCMS m / z = 437.05 (M+H), t R = 1.41 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Phenomenex Kinetex 2.6 μ XB-C18 100 A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% acetonitrile, 1.8 min - 1.85 min 100% - 2% acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min.

[0639] ​ ​

[0640]

[0641] A solution of 4-nitrobenzene phosphorodichloridate (503 mg, 1.97 mmol) in dichloromethane (20 mL) was added dropwise to a solution of 3-morpholinopropyl L-alaninate hydrochloride (496 mg, 1.97 mmol) in dichloromethane (20 mL) at 0 °C over 10 min. After the addition was complete, triethylamine (0.55 mL, 3.93 mmol) was added dropwise. After 90 min, phenol (185 mg, 1.97 mmol) and triethylamine (0.28 mL, 1.97 mmol) were added sequentially at 0 °C, and the resulting mixture was allowed to warm to RT. After 30 min, the reaction mixture was washed with water (2 x 50 mL) and brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude residue was purified by silica gel chromatography eluting with 20-100% ethyl acetate in hexanes to give the product. 1 H NMR (400 MHz, Methanol-d4) δ 8.32 - 8.24 (m, 2H), 7.51 - 7.39 (m, 2H), 7.24 - 7.12 (m, 2H), 6.97 - 6.90 (m, 2H), 4.94 (m, 1H), 4.12 - 4.07 (m, 2H), 4.05 - 3.93 (m, 1H), 3.76 - 3.68 (m, 2H), 3.41 (d, J = 0.5 Hz, 3H), 1.32 (td, J = 7.1, 1.2 Hz, 3H), 1.19 (dt, J = 6.3, 2.0 Hz, 6H). 31 P NMR (162 MHz, Acetonitrile-d3) δ -2.12, -2.22. LCMS: MS m / z = 494.35 [M+l], t R = 1.03 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Kinetex 2.6 μ XB-C18 100A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, Water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% Acetonitrile, 1.8 min - 1.85 min 100% - 2% Acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min.

[0642] ​

[0643]

[0644] (R)-tetrahydrofuran-3-yl-L-alaninate. To a mixture of N-Cbz-L-alanine (3.31 g, 14.83 mmol), (R)-THF-3-ol (1.0 mL, 12.34 mmol), and EDCI (2.49 g, 16.04 mmol) in acetonitrile (20 mL) was added DMAP (2.26 g, 18.51 mmol). The mixture was then stirred at room temperature for 15 hours, then diluted with EtOAc, washed with brine, dried over sodium sulfate, and concentrated in vacuo. The obtained residue was purified by silica gel chromatography (0% to 50% EtOAc in hexanes, 35 min run) to give Cbz-L-alanine ester (2.78 g), which was dissolved in THF (20 mL) and 20% Pd(OH)2(433 mg, 0.617 mmol) was added at room temperature. The resulting mixture was stirred under a hydrogen atmosphere at room temperature for 4.5 hours, filtered, concentrated in vacuo, and dried under high vacuum to give the product. 1 H NMR (400 MHz, Chloroform-d) δ 5.32 (ddt, J = 6.5, 4.3, 1.9 Hz, 1H), 3.98 - 3.78 (m, 4H), 3.56 (q, J = 7.0 Hz, 1H), 2.19 (dtd, J = 13.7, 8.4, 6.4 Hz, 1H), 2.05 - 1.92 (m, 1H), 1.79 (s, 2H), 1.34 (d, J = 7.0 Hz, 3H). LCMS m / z = 159.92 (M+H), t R = 0.21 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Chromatography column: Phenomenex Kinetex 2.6 μ XB-C18 100A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% acetonitrile, 1.8 min - 1.85 min 100% - 2% acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min.

[0645]

[0646] (R)-tetrahydrofuran-3-yl ((4-nitrophenoxy)(phenoxy)phosphoryl)-L-alaninate. To a solution of (R)-tetrahydrofuran-3-yl-L-alaninate (1.66 g, 10.44 mmol) in DCM (40 mL) at -78 °C was added phenyl dichlorophosphate (1.56 mL, 10.44 mmol) in one portion rapidly. Then triethylamine (1.45 mL, 10.44 mmol) was added at -78 °C within 5 min. After removing the dry ice bath, the resulting mixture was stirred for 30 min and cooled to -78 °C. p-Nitrophenol (1.45 g, 10.44 mmol) was added in one portion and triethylamine (1.45 mL, 10.44 mmol) was added at -78 °C within 5 min. After removing the dry ice bath, the resulting mixture was stirred for 2 hours. After dilution with DCM, the mixture was washed with brine, concentrated in vacuo and the resulting residue was purified by silica gel column chromatography (0% to 100% EtOAc in hexanes) to give the product. 1 HNMR (400 MHz, Chloroform-d) δ 8.22 (m, 2H), 7.43 - 7.31 (m, 4H), 7.25 - 7.14 (m, 3H), 5.29 (m, 1H), 4.21 - 4.10 (m, 1H), 3.93 - 3.79 (m, 4H), 3.79 - 3.71 (m, 1H), 2.17 (m, 1H), 1.97 - 1.85 (m, 1H), 1.44 - 1.37 (m, 3H). 31 P NMR (162 MHz, Chloroform-d) δ -3.24, -3.26. LCMS m / z = 437.02 (M+H), t R = 1.42 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Phenomenex Kinetex 2.6 μ XB-C18 100A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, Water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% Acetonitrile, 1.8 min - 1.85 min 100% - 2% Acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min.

[0647] ​

[0648]

[0649] Under an argon atmosphere, to a solution of phenol (4.70 mg, 50.0 mmol) in TBME (72 mL) at -78 °C was added phosphorous oxychloride (5.08 mL, 50.0 mmol) followed by triethylamine (6.97 mL, 50.0 mmol). The reaction mixture was then allowed to warm to RT. After 1 h, the resulting mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was dissolved in dichloromethane (72 mL) and L-alanine methyl ester hydrochloride (6.97 mg, 50.0 mmol) was added. The resulting suspension was cooled to -78 °C and triethylamine (13.9 mL, 100 mmol) was added dropwise. The reaction mixture was then allowed to warm to RT. After 16 h, the reaction mixture was concentrated under reduced pressure and TBME (100 mL) was added to the residue. The resulting white solid was removed by vacuum filtration and the filtrate was concentrated under reduced pressure to give the product which was used directly in the next step. 1 H NMR (400 MHz, Chloroform-d1) δ 7.45 - 7.12 (m, 5H), 4.67 - 4.44 (m, 1H), 4.44 - 4.24 (m, 1H), 3.81 (s, 1.5H), 3.78 (s, 1.5H), 1.53 (app t, J = 6.8 Hz, 3H). 31 P NMR (162 MHz, Chloroform-d1) δ 64.78 (s), 64.63 (s).

[0650] ​ Phosphoryl)-L-alanine cyclohexyl ester and (((S)-1-cyclohexyloxy-1-oxopropan-2-yl)amino)(4-nitrophenoxy) Phosphoryl)-L-alanine cyclohexyl ester

[0651]

[0652] To a solution of (S)-l-(cyclohexyloxy)-l-oxopropane-2-ium chloride intermediate 11 (680 mg, 3.27 mmol) in THF (10 mL) was added 4-nitrophenylphosphonic dichloride (838 mg, 3.27 mmol) in one portion. The resulting mixture was cooled in an ice bath and a solution of triethylamine (1.0 mL, 6.54 mmol) in THF (2 mL) was added over 30 min. The resulting mixture was stirred in an ice bath for 1.5 h, (S)-l-(2-ethylbutoxy)-l-oxopropane-2-ium chloride (687 mg, 3.27 mmol) was added in one portion, and a solution of triethylamine (1.0 mL, 6.54 mmol) in THF (2 mL) was added over 30 min in an ice bath. The resulting mixture was stirred in an ice bath for 1.5 h, diluted with EtOAc, washed with water and brine, concentrated in vacuo, and the resulting residue was purified by preparative HPLC (Phenomenex Gemini-NX 10 μ C18 110 A 250 x 30 mm column, 0% - 100% acetonitrile / water gradient, 25 min run) to give the product. 1 H NMR (400 MHz, Chloroform-d) δ 8.20 (m, 2H), 7.38 (m, 2H), 4.77 (m, 1H), 4.15 - 3.91 (m, 4H), 3.60 (m, 2H), 1.91 - 1.77 (m, 2H), 1.75 - 1.67 (m, 2H), 1.51 (m, 2H), 1.45 - 1.23 (m, 15H), 0.88 (m, 6H). 31 P NMR (162 MHz, Chloroform-d) δ 8.04. LCMS: MS m / z = 528.10 [M+l].

[0653] Intermediate 60. 4-nitrophenyl-N,N'-cyclohexyl L-alanine diaminophosphoramidate

[0654]

[0655] (S)-2-Aminopropanoic acid cyclohexyl ester hydrochloride. L-alanine (891 mg, 10 mmol) was combined with cyclohexanol (10 mL). Trimethylchlorosilane (12.7 mL, 100 mmol) was added dropwise and stirred for 20 min. The reaction mixture was heated to 60 °C and stirred for 16 h. The reaction was concentrated under reduced pressure and azeotroped with toluene (5x) to give an oil. Hexane (100 mL) was added and stirred for 15 h to give a solid which was collected, washed with hexane (100 mL) and dried under high vacuum to give the product. 1H NMR (400 MHz, DMSO-d6) δ 8.45 (s, 3H), 4.77 (tt, J = 8.4, 3.7 Hz, 1H), 4.02 (q, J = 7.2 Hz, 1H), 1.71 (m, 4H), 1.53 - 1.17 (m, 9H).

[0656]

[0657] 4-Nitrophenyl-N,N'-cyclohexyl L-alanine diaminophosphoramide. 4-Nitrophenylphosphorodichloridate (256 mg, 1 mmol) was dissolved in anhydrous dichloromethane (10 mL) and stirred in an ice bath under atmospheric nitrogen. (S)-2-Aminopropionic acid cyclohexyl ester hydrochloride (415 mg, 2 mmol) was added in one portion. Triethylamine (698 μL, 5 mmol) was added dropwise and stirred for 2 hours. The reaction was diluted with dichloromethane (15 mL) and washed with 2% aqueous citric acid (20 mL). The organics were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified via Si02column chromatography (12 g Si02Combiflash HP Gold column, 0% - 50% ethyl acetate / hexanes) to give the product. 1 H NMR (400 MHz, DMSO-d6) δ 8.30 - 8.13 (m, 2H), 7.49 - 7.27 (m, 2H), 5.50 (m, 2H), 4.62 (m, 2H), 3.85 (m, 2H), 1.67 (m, 8H), 1.51 - 1.18 (m, 18H). 31 P NMR (162 MHz, DMSO-d6) δ 9.50. MS m / z = 526.0 [M+1], 524.1 [M-1].

[0658] Intermediate 61. 4-nitrophenyl-N,N'-isopropyl L-alanine diaminophosphoramidate

[0659]

[0660] To a solution of L-alanine isopropyl ester HC1 salt (1.97 g, 11.72 mmol) in DCM (20 mL) was added 4-nitrophenylphosphorodichloridate (1.5 g, 5.86 mmol) in one portion. The resulting mixture was cooled to about 0 °C and triethylamine (2.37 g, 23.44 mmol) was added dropwise. After the ice bath was removed, the resulting mixture was stirred for about 30 min and stirred overnight. The reaction mixture was then diluted with EtOAc, washed with water and brine, the organic solvent was concentrated in vacuo and the resulting residue was purified by silica gel column chromatography eluting with 0% - 100% ethyl acetate in hexanes to give the product. LCMS: MS m / z = 445.96 [M+1].

[0661] Intermediate 62. 4-nitrophenyl-N,N'-cyclobutylmethyl L-alanine diaminophosphoramidate

[0662]

[0663] To a solution of L-alanine cyclobutylmethyl ester HC1 salt (1.51 g, 7.8 mmol) in DCM (20 mL) was added 4-nitrobenzenedichloride (1 g, 3.9 mmol) in one portion. The resulting mixture was cooled to 0 °C and triethylamine (1.58 g, 15.6 mmol) was added dropwise. After the ice bath was removed, the resulting mixture was stirred for 30 min and stirred overnight. The reaction mixture was then diluted with EtOAc, washed with water and brine, the organic solvent was concentrated in vacuo and the resulting residue was purified by silica gel column chromatography eluting with 0-100% ethyl acetate in hexanes to afford the product. LCMS: MS m / z = 497.98 [M+1].

[0664] Intermediate 63. (1r,4S)-4-((tert-butoxycarbonyl)amino)cyclohexyl ((4-nitrophenoxy)(phenoxy) phosphoramidate Phosphoryl)-L-alanine ester

[0665]

[0666] (1r,4S)-4-((tert-butoxycarbonyl)amino)cyclohexyl ((benzyloxy)carbonyl)-L-alanine ester. At RT, 4-dimethylaminopyridine (2.84 g, 23 mmol) was added to a solution of tert-butyl ((1r,4r)-4-hydroxycyclohexyl)carbamate (4.00 g, 19.0 mmol) and ((benzyloxy)carbonyl)-L-alanine (4.98 g, 22.0 mmol) and EDCI (3.13 g, 20.0 mmol) in acetonitrile (100 mL). After 4 h, the reaction mixture was diluted with dichloromethane (200 mL), washed with brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude residue was purified by silica gel chromatography eluting with 0-50% ethyl acetate in hexanes to afford the product. 1 H NMR (400 MHz, CDC13) δ 7.40 - 7.28 (m, 5H), 5.29 (br d, J = 7.7 Hz, 1H), 5.10 (s, 2H), 4.78 - 4.60 (m, 1H), 4.47 - 4.19 (m, 2H), 3.45 (s, 1H), 2.08 - 1.89 (m, 4H), 1.54 - 1.34 (m, 14H), 1.28 - 1.16 (m, 2H). LCMS: MS m / z = 420.99 [M+1].

[0667]

[0668] (1r,4S)-4-((tert-butoxycarbonyl)amino)cyclohexyl ((4-nitrophenoxy)(phenoxy)phosphoryl)-L-alaninate. Under an argon atmosphere, a hydrogen balloon was added to a flask containing (1r,4S)-4-((tert-butoxycarbonyl)amino)cyclohexyl ((benzyloxy)carbonyl)-L-alaninate (1.96 g, 4.66 mmol) and palladium on carbon (10 wt%, 2.0 g) in tetrahydrofuran (50 mL) at RT. The vessel was evacuated and refilled with a hydrogen atmosphere (3x) and the reaction mixture was stirred vigorously. After 1.5 h, the reaction mixture was filtered through a pad of celite and the filtrate was concentrated under reduced pressure to give the crude Cbz deprotected material. The crude residue was dissolved in dichloromethane (23 mL) and the resulting mixture was cooled to 0 °C. Phenyl dichlorophosphate (0.70 mL, 4.7 mmol) and triethylamine (0.66 mL, 4.7 mmol) were added sequentially. After 1 h, 4-nitrophenol (660 mg, 4.74 mmol) and triethylamine (0.66 mL, 4.7 mmol) were added. After 1.5 h, the reaction mixture was diluted with dichloromethane (50 mL), washed with saturated aqueous sodium bicarbonate (50 mL) and brine (50 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude residue was purified by silica gel chromatography eluting with 0% - 100% ethyl acetate in hexanes to give the product. 1 H NMR (400 MHz, Chloroform-d1) δ 8.26 - 8.18 (m, 2H), 7.43 - 7.30 (m, 4H), 7.25 - 7.17 (m, 3H), 4.77 - 4.58 (m, 1H), 4.40 (br s, 1H), 4.18 - 3.99 (m, 1H), 3.93 - 3.80 (m, 1H), 3.44 (br s, 1H), 2.07 - 1.87 (m, 4H), 1.52 - 1.36 (m, 14H), 1.30 - 1.16 (m, 2H). 31 P NMR (162 MHz, Chloroform-d1) δ -3.15 (s). LCMS: MS m / z = 563.88 [M+1].

[0669] Intermediate 64. ((1r,4S)-4-((tert-butoxycarbonyl)amino)cyclohexyl)methyl ((4-nitrophenoxy)(phenyl) phosphoryl)-L-alanine ester Phosphoryl)-L-alanine ester

[0670] Method 1 .

[0671]

[0672] ((1 r,4S)-4-((tert-butoxycarbonyl)amino)cyclohexyl)methyl ((benzyloxy)carbonyl)- L-alaninate. Cbz-L-alanine (223 mg, 1.00 mmol) was dissolved in anhydrous MeCN (10 mL). Trans-1-(Boc-amino)-4-(hydroxymethyl)cyclohexane (229 mg, 1.00 mmol) and EDCI (230 mg, 1.2 mmol) were added to the reaction, which was then stirred for 25 min. DMAP (122 mg, 1 mmol) was added in one portion, and the reaction was stirred for 4 h. The reaction mixture was diluted with ethyl acetate (15 mL) and washed with 5% aqueous citric acid (2 x 5 mL), followed by brine (10 mL). The organic extract was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude residue was purified via SiO2column chromatography (12 g SiO2Combiflash HP Gold column, 0% - 40% ethyl acetate / hexanes). Fractions containing the desired product were combined and concentrated under reduced pressure to yield the product. 1 HNMR (400 MHz, Chloroform-d) δ 7.41 - 7.27 (m, 5H), 5.29 (d, J = 7.6 Hz, 1H), 5.11 (s, 2H), 4.47 - 4.24 (m, 2H), 3.96 (d, J = 6.6 Hz, 2H), 3.37 (bs, 1H), 2.03 (m, 2H), 1.78 (m, 2H), 1.58 (m, 2H), 1.44 (m, 12H), 1.10 (m, 4H).

[0673]

[0674] ((1 r,4S)-4-((tert-butoxycarbonyl)amino)cyclohexyl)methyl ((4- nitrophenoxy)(phenoxy)phosphoryl)-L-alaninate. ((1 r,4S)-4-((tert- butoxycarbonyl)amino)cyclohexyl)methyl ((benzyloxy)carbonyl)-L-alaninate (348 mg, 0.800 mmol) was dissolved in 12 mL of anhydrous tetrahydrofuran. Degussa 10% palladium on carbon (25 mg) was added to the reaction, which was then stirred under an atmosphere of hydrogen gas for 3 hours. The palladium on carbon was filtered off and the filtrate was used in the next reaction without further purification. Phenyl dichlorophosphate (119 μL, 0.800 mmol) was dissolved in 15 mL of anhydrous dichloromethane and stirred in an ice bath under an atmosphere of nitrogen gas. The above filtrate was then added dropwise to the reaction solution, which was then stirred for 30 min. Triethylamine (120 μL, 0.88 mmol) was added dropwise and stirred for 1 hour. p-Nitrophenol (100 mg, 0.72 mmol) was added in one portion. Triethylamine (123 μL, 0.88 mmol) was added dropwise and the reaction mixture was stirred at RT for 2 hours. The reaction mixture was then diluted with dichloromethane (10 mL) and washed with water (3 x 10 mL). The organic extract was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude residue was purified via Si02column chromatography (12 g Si02Combiflash HP Gold column, 0% - 40% ethyl acetate / hexanes). Fractions containing the desired product were combined and concentrated under reduced pressure to yield the product. 1 H NMR (400 MHz, Chloroform-d) δ 8.27 - 8.18 (m, 2H), 7.44 - 7.30 (m, 4H), 7.27 - 7.17 (m, 3H), 4.35 (s, 1H), 4.22 - 4.06 (m, 1H), 3.99 - 3.88 (m, 2H), 3.85 (t, J = 10.6 Hz, 1H), 3.36 (s, 1H), 2.03 (m, 2H), 1.75 (m, 2H), 1.57 (m, 2H), 1.48 - 1.36 (m, 12H), 1.15 - 0.98 (m, 4H). 31 P NMR (162 MHz, Chloroform-d) δ 3.12, 3.13. LCMS: MS m / z = 478.2 [M+l].

[0675] Method 2 .

[0676]

[0677] ((1 r,4S)-4-((tert-butoxycarbonyl)amino)cyclohexyl)methyl ((benzyloxy)carbonyl)- L-alaninate. To a solution of trans-1-((tert-butoxycarbonyl)amino)-4- (hydroxymethyl)cyclohexane (510 mg, 2.18 mmol), followed by N-(3- dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (509 g, 2.62 mmol) was added to Z-Ala-OH (489 g, 2.18 mmol) in acetonitrile (22 mL) at RT. After 30 min, 4- (dimethylamino)pyridine (267 mg, 2.18 mmol) was added. After 18 h, the reaction was diluted with ethyl acetate (100 mL) and the resulting mixture was washed with 10% aqueous citric acid (2 x 100 mL) and brine (100 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude residue was subjected to silica gel chromatography eluting with 0-50% methanol in ethyl acetate to afford the product. 1 H NMR (400 MHz, Chloroform-d) δ 7.41 - 7.29 (m, 5H), 5.28 (s, 1H), 5.11 (s, 2H), 4.46 - 4.27 (m, 2H), 3.96 (d, J = 6.6 Hz, 2H), 3.37 (s, 1H), 2.03 (s, 2H), 1.78 (s, 2H), 1.56 (s, 2H), 1.44 (s, 9H), 1.42 (d, J = 7.2 Hz, 3H), 1.08 (t, J = 9.7 Hz, 4H). LCMS: MS m / z = 434.87 [M+l], t R = 1.21 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Kinetex 2.6 μ XB-C18 100A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, Water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% Acetonitrile, 1.8 min - 1.85 min 100% - 2% Acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min. HPLC: t R = 5.96 min; HPLC system: Agilent 1100 series; Column: Gemini 5 μ C18 110A, 50 x 4.6 mm; Solvents: Acetonitrile with 0.1% TFA, Water with 0.1% TFA; Gradient: 0 min - 9.0 min 2% - 95% ACN, 9.0 min - 10.0 min 95% ACN, at a rate of 2 mL / min.

[0678]

[0679] ((1 r,4S)-4-((tert-butoxycarbonyl)amino)cyclohexyl)methyl L-alaninate. Palladium on carbon (198 mg, 10 wt%) was added to a solution of ((1 r,4S)-4-((tert- butoxycarbonyl)amino)cyclohexyl)methyl ((benzyloxy)carbonyl)-L-alaninate (719 mg, 1.65 mmol) in tetrahydrofuran (24 mL) purged with argon. The mixture was then purged with hydrogen gas and stirred at RT. After 1 h, the mixture was filtered through celite, the filter was rinsed with tetrahydrofuran, and the volatiles were removed under reduced pressure to give the product. 1 H NMR (400 MHz, Chloroform-d) δ 4.38 (s, 1H), 4.02 - 3.85 (m, 2H), 3.55 (q, J = 7.0 Hz, 1H), 3.38 (s, 1H), 2.04 (d, J = 7.1 Hz, 2H), 1.83 - 1.73 (m, 2H), 1.63 (s, 2H), 1.44 (s, 10H), 1.34 (d, J = 7.0 Hz, 3H), 1.09 (t, J = 10.0 Hz, 4H). LCMS: MS m / z = 300.93 [M+l], t R = 0.65 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Chromatography column: Kinetex 2.6 μ XB-C18 100A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% acetonitrile, 1.8 min - 1.85 min 100% - 2% acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min.

[0680]

[0681] ((1 r,4S)-4-((tert-butoxycarbonyl)amino)cyclohexyl)methyl ((4- nitrophenoxy)(phenoxy)phosphoryl)-L-alaninate. To a solution of ((1 r,4S)-4-((tert- butoxycarbonyl)amino)cyclohexyl)methyl L-alaninate (553 mg, 1.65 mmol) in tetrahydrofuran (24 mL) was added slowly a solution of phenyl dichlorophosphate (247 μL, 1.65 mmol) in dichloromethane (30 mL) at 0 °C over 15 min. After the addition was complete, triethylamine (0.26 mL, 1.82 mmol) was added dropwise. After 1 h, 4-nitrophenol (240 mg, 1.65 mmol) and triethylamine (0.26 mL, 1.82 mmol) were added sequentially at 0 °C and the resulting mixture was allowed to warm to RT. After 1 h, the reaction mixture was diluted with dichloromethane (50 mL) and washed with water (3 x 75 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude residue was purified by silica gel chromatography eluting with 0-100% ethyl acetate in hexanes to give the product. 1 H NMR (400 MHz, Chloroform-d) δ 8.23 (ddd, J = 9.3, 1.3, 0.6 Hz, 2H), 7.44 - 7.31 (m, 4H), 7.25 - 7.16 (m, 3H), 4.36 (s, 1H), 4.22 - 4.06 (m, 1H), 3.96 - 3.90 (m, 2H), 3.84 (t, J = 10.6 Hz, 1H), 3.36 (s, 1H), 2.02 (s, 2H), 1.83 - 1.68 (m, 2H), 1.57 (s, 2H), 1.44 (s, 9H), 1.41 (dd, J = 7.1, 3.2 Hz, 3H), 1.06 (t, J = 9.6 Hz, 3H). 31 P NMR (162 MHz, Chloroform-d) δ -3.13 (d, J = 2.9 Hz). LCMS: MS m / z = 577.8 [M+l], t R = 1.28 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Kinetex 2.6 μ XB-C18 100A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, Water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% Acetonitrile, 1.8 min - 1.85 min 100% - 2% Acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min. HPLC: t R= 6.35 min; HPLC system: Agilent 1100 series; column: Gemini 5μ C18 110A, 50 x 4.6 mm; solvents: acetonitrile with 0.1% TFA, water with 0.1% TFA; gradient: 0 min - 9.0 min 2% - 95% ACN, 9.0 min - 10.0 min 95% ACN, at a rate of 2 mL / min.

[0682] Intermediate 65. 4-nitrophenyl-N,N'-butanoyl L-alanine diaminophosphoramidate

[0683]

[0684] (tert-Butoxycarbonyl)-L-alanine butyl ester. Boc-L-alanine (380 mg, 2.0 mmol) was dissolved in anhydrous MeCN (10 mL). 1-Butanol (920 μL, 10.0 mmol) and EDCI (460 mg, 2.4 mmol) were added to the reaction, which was then stirred for 15 min. DMAP (240 mg, 2.0 mmol) was added in one portion and the reaction was stirred for 14 h. The reaction mixture was diluted with ethyl acetate (15 mL) and washed with saturated aqueous sodium bicarbonate solution (2 x 10 mL) followed by brine (5 mL). The organic extract was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude residue was purified by SiO2column chromatography (12 g SiO2Combiflash HP Gold column, 0% - 20% ethyl acetate / hexanes). Fractions containing the desired product were combined and concentrated under reduced pressure to yield the product. 1 H NMR (400 MHz, Chloroform-d) δ 5.04 (m, 1H), 4.29 (m, 1H), 4.18 - 4.07 (m, 2H), 1.67 - 1.59 (m, 2H), 1.44 (s, 9H), 1.38 (m, 5H), 0.93 (t, J = 7.4 Hz, 3H).

[0685]

[0686] 4-nitrophenyl-N,N'-butanoyl L-alanine diaminophosphoramidate. (tert- Butoxycarbonyl)-L-alanine butyl ester (291 mg, 1.18 mmol) was dissolved in 7 mL of 4M HC1 in dioxane and stirred for 1 hour. The reaction mixture was concentrated under reduced pressure to give an oil which was then dissolved in anhydrous dichloromethane (10 mL) and stirred in an ice bath under atmospheric nitrogen. 4-nitrophenyl phosphorodichloridate (152 mg, 0.59 mmol) was added in one portion and the reaction was stirred for 10 min. Triethylamine (270 μL, 1.95 mmol) was dissolved in 1 mL of anhydrous dichloromethane and added dropwise to the reaction solution. The reaction mixture was stirred for 1 hour. Triethylamine (270 μL, 1.95 mmol) was dissolved in 700 μL of anhydrous dichloromethane and added dropwise to the reaction. The reaction mixture was stirred at RT for 16 hours. The reaction mixture was diluted with dichloromethane (15 mL) and washed with water (3 x 20 mL). The organic extract was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude residue was purified via Si02column chromatography (12 g Si02Combiflash HP Gold chromatography column, 0% - 50% ethyl acetate / hexanes). Fractions containing the desired product were combined and concentrated under reduced pressure to give the product. 1 H NMR (400 MHz, Chloroform-d) δ 8.27 - 8.15 (m, 2H), 7.43 - 7.34 (m, 2H), 4.19 - 3.98 (m, 5H), 3.80 - 3.61 (m, 1H), 3.58 (m, 2H), 1.67 - 1.59 (m, 4H), 1.45 - 1.30 (m, 10H), 0.93 (m, 6H). 31 PNMR (162 MHz, Chloroform-d) δ 7.93. LCMS: MS m / z = 474.0 [M+l].

[0687] Intermediate 66. methyl (2S)-2-(((benzyloxy)carbonyl)amino)-3-(4-(((((S)-1- isopropoxy-1-oxopropan-2-yl)amino)(4-nitrophenoxy) phosphoryl)oxy)phenyl)propanoate Intermediate 67. 2-morpholinoethyl ((4-nitrophenoxy)(phenoxy) phosphoryl)-L-alanine ester

[0688]

[0689] A solution of 4-nitrobenzene phosphorodichloridate (504 mg, 1.97 mmol) in dichloromethane (20 mL) was added dropwise to a solution of L-alanine isopropyl ester hydrochloride (330 mg, 1.97 mmol) in dichloromethane (20 mL) at 0 °C over 10 min. After the addition was complete, triethylamine (0.55 mL, 3.93 mmol) was added dropwise. After 60 min, N- benzyloxycarbonyl-L-tyrosine methyl ester (649 mg, 1.97 mmol) and triethylamine (0.28 mL, 1.97 mmol) were added sequentially at 0 °C, then the resulting mixture was allowed to warm to RT. After 30 min, the reaction mixture was washed with water (2 x 50 mL) and brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude residue was purified by silica gel chromatography eluting with 20-100% ethyl acetate in hexanes to give the product. 1 H NMR (400 MHz, Methanol-d4) δ 8.32 - 8.22 (m, 2H), 7.49 - 7.37 (m, 2H), 7.35 - 7.13 (m, 9H), 5.02 (s, 2H), 4.93 (pd, J = 6.3, 1.1 Hz, 1H), 4.43 (dd, J = 9.4, 5.2 Hz, 1H), 4.00 (dtd, J = 10.1, 7.7, 6.5 Hz, 1H), 3.70 (s, 3H), 3.15 (dd, J = 14.0, 5.4 Hz, 1H), 2.93 (dd, J = 13.9, 9.6 Hz, 1H), 1.32 (td, J = 7.2, 1.2 Hz, 3H), 1.20 - 1.16 (m, 6H). 31 P NMR (162 MHz, Methanol-d4) δ -1.26, -1.49. LCMS: MS m / z = 644.11 [M+1], t R = 1.56 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Kinetex 2.6 μ XB-C18 100A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, Water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% Acetonitrile, 1.8 min - 1.85 min 100% - 2% Acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min. HPLC: t R= 6.21 min; HPLC system: Agilent 1100 series; column: Gemini 5μ C18 110A, 50 x 4.6 mm; solvents: acetonitrile with 0.1% TFA, water with 0.1% TFA; gradient: 0 min - 9.0 min 2% - 95% ACN, 9.0 min - 10.0 min 95% ACN, rate 2 mL / min.

[0690] Intermediate 68. 2-(diisopropylamino)ethyl ((4-nitrophenoxy)(phenoxy) phosphoryl)-L-alanine ester

[0691]

[0692] A solution of 4-nitrobenzene phosphorodichloridate (505 mg, 1.97 mmol) in dichloromethane (20 mL) was added dropwise to a solution of 2-morpholinoethyl L-alaninate hydrochloride (496 mg, 1.97 mmol) in dichloromethane (20 mL) at 0 °C over 10 min. After the addition was complete, triethylamine (0.55 mL, 3.93 mmol) was added dropwise. After 90 min, phenol (185 mg, 1.97 mmol) and triethylamine (0.28 mL, 1.97 mmol) were added sequentially at 0 °C and the resulting mixture was allowed to warm to RT. After 30 min, the reaction mixture was washed with water (2 x 50 mL) and brine (50 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude residue was purified by silica gel chromatography eluting with 20% - 100% ethyl acetate in hexanes to give the product. 1 H NMR (400 MHz, CDC13) δ 8.28 - 8.14 (m, 2H), 7.41 - 7.29 (m, 4H), 7.24 - 7.16 (m, 4H), 6.87 - 6.81 (m, 1H), 4.14 - 4.04 (bs, 2H), 2.61 - 2.57 (bs, 4H), 2.45 - 3.40 (bs, 4H), 1.42 (dt, J=6.3, 2.0 Hz, 6H). 31 P NMR (162 MHz, CDC13) δ -2.70. LCMS: MS m / z = 480.27 [M+l], t R= 0.96 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Chromatography column: Kinetex 2.6 u XB-C18 100A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% acetonitrile, 1.8 min - 1.85 min 100% - 2% acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 uL / min. HPLC: t R = 5.23 min; HPLC system: Agilent 1100 series; Chromatography column: Kinetx 2.6u 100A C18, 100 mm x 4.6 mm; Solvents: Acetonitrile with 0.1% TFA, water with 0.1% TFA; Gradient: 0 min - 8.5 min 2% - 98% ACN, 8.5 min - 10.0 min 98% ACN, at a rate of 1.5 mL / min.

[0693] Intermediate 69. isopropyl (2S)-2-(((benzyloxy)carbonyl)amino)-3-(4-(((((S)-1- methoxy-1-oxopropan-2-yl)amino)(4-nitrophenoxy) phosphoryl)oxy)phenyl)propanoate Intermediate 70. isopropyl ((2-(methylthio)ethoxy)(4-nitrophenoxy) phosphoryl)-L-alanine ester

[0694]

[0695] 2-(Diisopropylamino)ethyl ((benzyloxy)carbonyl)-L-alaninate. At RT, N-(3- dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (2.06 g, 10.8 mmol) was added to a solution of Z-Ala-OH (2.00 g, 8.96 mmol) and 2- (diisopropylamino)ethanol (3.2 mL, 17.9 mmol) in acetonitrile (125 mL). After 10 min, 4-(dimethylamino)pyridine (1.09 g, 8.96 mmol) was added. After 2 days, the reaction mixture was concentrated to half volume, the mixture was diluted with ethyl acetate (100 mL), and the resulting mixture was washed with saturated aqueous sodium carbonate (100 mL) and brine (100 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude residue was subjected to silica gel chromatography eluting with 0% - 20% methanol in ethyl acetate to give the product. 1 H NMR (400 MHz, Acetonitrile-d3) δ 7.48 - 7.23 (m, 5H), 5.96 (s, 1H), 5.07 (s, 2H), 4.30 - 4.00 (m, 3H), 2.28 (t, J = 7.1 Hz, 2H), 2.14 (s, 6H), 1.73 (p, J = 6.9 Hz, 2H), 1.34 (d, J = 7.3 Hz, 3H). LCMS: MS m / z = 351.26 [M+1], tR = 1.05 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Kinetex 2.6 μ XB-C18 100A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% acetonitrile, 1.8 min - 1.85 min 100% - 2% acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min. HPLC: t R = 3.10 min; HPLC system: Agilent 1100 series; Column: Gemini 5 μ C18 110A, 50 x 4.6 mm; Solvents: Acetonitrile with 0.1% TFA, water with 0.1% TFA; Gradient: 0 min - 9.0 min 2% - 95% ACN, 9.0 min - 10.0 min 95% ACN, at a rate of 2 mL / min.

[0696]

[0697] 2-(Diisopropylamino)ethyl L-alaninate. Palladium on carbon (587 mg, 10 wt%) was added to a solution of 2-(diisopropylamino)ethyl ((benzyloxy)carbonyl)-L-alaninate (1.93 g, 5.52 mmol) in ethanol (50 mL) purged with argon. The mixture was then purged with hydrogen gas and stirred at RT. After 18 h, the mixture was filtered through celite, the filter was rinsed with ethyl acetate, and the volatiles were removed under reduced pressure to give the product. 1 H NMR (400 MHz, Acetonitrile-d3) δ 4.06 - 3.90 (m, 2H), 3.43 (q, J = 7.0 Hz, 1H), 3.01 (hept, J = 6.5 Hz, 2H), 2.65 (t, J = 6.9 Hz, 2H), 1.22 (d, J = 7.0 Hz, 3H), 0.99 (d, J = 6.6 Hz, 12H). LCMS: MS m / z = 217.01 [M+l], t R= 0.17 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Chromatography column: Kinetex 2.6 μ XB-C18 100A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% acetonitrile, 1.8 min - 1.85 min 100% - 2% acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min.

[0698]

[0699] 2-(Diisopropylamino)ethyl ((4-nitrophenoxy)(phenoxy)phosphoryl)-L-alaninate. A solution of 2-(diisopropylamino)ethyl L-alaninate (511 mg, 2.43 mmol) in tetrahydrofuran (7 mL) was added dropwise to a solution of phenyl dichlorophosphate (0.36 mL, 2.43 mmol) in tetrahydrofuran (25 mL) at 0 °C over 15 min. After the addition was complete, triethylamine (0.36 mL, 2.43 mmol) was added dropwise. After 90 min, 4-nitrophenol (337 mg, 2.43 mmol) and triethylamine (1.0 mL, 7.16 mmol) were added sequentially at 0 °C, and the resulting mixture was allowed to warm to RT. After 17 h, the reaction mixture was diluted with ethyl acetate (100 mL) and washed with water (2 x 100 mL) and brine (100 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the product. 1 H NMR (400 MHz, Acetonitrile-d3) δ 8.29 - 8.18 (m, 2H), 7.49 - 7.35 (m, 4H), 7.30 - 7.21 (m, 3H), 4.71 - 4.52 (m, 1H), 4.12 - 3.99 (m, 2H), 4.00 - 3.83 (m, 3H), 3.06 - 2.86 (m, 2H), 2.56 (td, J = 7.0, 3.8 Hz, 2H), 1.31 (ddd, J = 7.1, 4.7, 1.1 Hz, 4H), 0.94 (d, J = 6.5 Hz, 13H). 31 P NMR (162 MHz, Acetonitrile-d3) δ -2.15, -2.30. LCMS: MS m / z = 494.25 [M+l], t R= 1.27 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Chromatography column: Kinetex 2.6 μ XB-C18 100A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% acetonitrile, 1.8 min - 1.85 min 100% - 2% acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min. HPLC: t R = 3.97 min; HPLC system: Agilent 1100 series; Chromatography column: Gemini 5 μ C18 110A, 50 x 4.6 mm; Solvents: Acetonitrile with 0.1% TFA, water with 0.1% TFA; Gradient: 0 min - 9.0 min 2% - 95% ACN, 9.0 min - 10.0 min 95% ACN, at a rate of 2 mL / min.

[0700] Intermediate 71. isopropyl ((2-methoxyethoxy)(4-nitrophenoxy) phosphoryl)-L-alanine ester Intermediate 72. isopropyl ((2-(methylsulfonyl)ethoxy)(4-nitrophenoxy) phosphoryl)-L-alanine ester

[0701]

[0702] ((Benzoyl)oxy)-L-tyrosine isopropyl ester. Benzyl chloroformate (0.94 mL, 6.58 mmol) was added dropwise to a mixture of L-tyrosine isopropyl ester (1.0 g, 4.48 mmol) in acetone (4.5 mL) and 7 wt% aqueous sodium carbonate (4.5 mL). After 2 h, the reaction mixture was diluted with ethyl acetate (25 mL) and the resulting mixture was washed with water (10 mL) and brine (10 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude residue was subjected to silica gel chromatography eluting with 0% - 100% ethyl acetate in hexanes to give the product. 1 H NMR (400 MHz, Methanol-d4) δ 7.39 - 7.20 (m, 5H), 7.06 - 6.97 (m, 2H), 6.74 - 6.62 (m, 2H), 5.05 (d, J = 2.6 Hz, 2H), 4.94 (p, J = 6.3 Hz, 1H), 4.31 (dd, J = 8.6, 6.1 Hz, 1H), 2.99 (dd, J = 13.9, 6.1 Hz, 1H), 2.84 (dd, J = 13.9, 8.6 Hz, 1H), 1.22 (d, J = 6.3 Hz, 3H), 1.14 (d, J = 6.3 Hz, 3H). LCMS: MS m / z = 357.87 [M+l], t R= 1.36 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Chromatography column: Kinetex 2.6 μ XB-C18 100 A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% acetonitrile, 1.8 min - 1.85 min 100% - 2% acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min. HPLC: t R = 5.19 min; HPLC system: Agilent 1100 series; Chromatography column: Gemini 5 μ C18 110 A, 50 x 4.6 mm; Solvents: Acetonitrile with 0.1% TFA, water with 0.1% TFA; Gradient: 0 min - 9.0 min 2% - 95% ACN, 9.0 min - 10.0 min 95% ACN, at a rate of 2 mL / min.

[0703]

[0704] (2S)-2-(((benzyloxy)carbonyl)amino)-3-(4-(((((S)-1-methoxy-1- oxopropan-2-yl)amino)(4-nitrophenoxy)phosphoryl)oxy)phenyl)propanoic acid isopropyl ester. A solution of L-alanine isopropyl ester hydrochloride (97.2 mg, 0.70 mmol) in dichloromethane (8.0 mL) was added to a solution of phosphorodichloridite of 4-nitrophenol (179.7 mg, 0.70 mmol) in dichloromethane (7.5 mL) at 0 °C. After the addition was complete, triethylamine (0.22 mL, 1.57 mmol) was added dropwise. After 60 min, a solution of ((benzyloxy)carbonyl)-L-tyrosine isopropyl ester (250.9 mg, 0.70 mmol) in dichloromethane (8.0 mL) and triethylamine (0.11 mL, 0.78 mmol) were added sequentially at 0 °C, and the resulting mixture was allowed to warm to RT. After 20 min, the reaction mixture was washed with water (2 x 20 mL) and brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude residue was purified by silica gel chromatography eluting with 0% - 100% ethyl acetate in hexanes to give the product. 1H NMR (400 MHz, Methanol-d4) δ 8.32 - 8.24 (m, 2H), 7.43 (ddd, J = 16.0, 9.2, 1.1 Hz, 2H), 7.36 - 7.09 (m, 9H), 5.03 (s, 2H), 4.97 (p, J = 6.2 Hz, 1H), 4.35 (d, J = 8.2 Hz, 1H), 4.14 - 3.95 (m, 1H), 3.62 (d, J = 4.5 Hz, 3H), 3.12 (dt, J = 12.6, 5.9 Hz, 1H), 2.92 (t, J = 11.6 Hz, 1H), 1.35 - 1.30 (m, 3H), 1.22 (d, J = 6.2 Hz, 3H), 1.16 (d, J = 6.2 Hz, 4H). 31 P NMR (162 MHz, Methanol-d4) δ -1.31, -1.52. LCMS: MS m / z = 644.07 [M+l], t R = 1.56 min; LC system: Thermo Accela 1250 UHPLC; MS system: Thermo LCQ Fleet; Column: Kinetex 2.6 μ XB-C18 100A, 50 x 3.0 mm; Solvents: Acetonitrile with 0.1% formic acid, water with 0.1% formic acid; Gradient: 0 min - 1.8 min 2% - 100% acetonitrile, 1.8 min - 1.85 min 100% - 2% acetonitrile, 1.85 min - 2.00 min 2% ACN, at a rate of 1800 μL / min. HPLC: t R = 6.17 min; HPLC system: Agilent 1100 series; Column: Gemini 5 μ C18 110A, 50 x 4.6 mm; Solvents: Acetonitrile with 0.1% TFA, water with 0.1% TFA; Gradient: 0 min - 9.0 min 2% - 95% ACN, 9.0 min - 10.0 min 95% ACN, at a rate of 2 mL / min.

[0705] Intermediate 73. 2-(2-ethoxyethoxy)ethyl ((perfluorophenoxy)(phenoxy) phosphoryl)-L-alanine ester single isomer

[0706]

[0707] Phosphorus dichloride, 4-nitrobenzene (512 mg, 2 mmol) was mixed with 10 mL of dry dichloromethane and stirred in an ice bath under atmospheric nitrogen. L-alanine isopropyl ester hydrochloride (335 mg, 2 mmol) was dissolved in dry dichloromethane (3 mL) and added dropwise to the reaction. The reaction mixture was stirred for 30 min. Triethylamine (927 μL, 6.6 mmol) was dissolved in dry dichloromethane (1 mL) and added dropwise to the reaction and the reaction stirred for 60 min. 2-(Methylthio)ethanol (74 μL, 2 mmol) was added in one portion and the reaction mixture stirred for 16 hours. The reaction mixture was diluted with dichloromethane (15 mL) and washed with water (3 x 20 mL). The organic extract was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude residue was purified by Si02column chromatography (12 g Si02Combiflash HP Gold column, 0% - 50% ethyl acetate / hexane). Fractions containing the desired product were combined and concentrated under reduced pressure to give the product. 1 H NMR (400 MHz, Chloroform-d) δ 8.27 - 8.18 (m, 2H), 7.44 - 7.33 (m, 2H), 5.02 (m, 1H), 4.33 - 4.21 (m, 2H), 4.07 - 3.94 (m, 1H), 3.70 (m, 1H), 2.84 - 2.73 (m, 2H), 2.14 (m, 3H), 1.40 (m, 3H), 1.29 - 1.19 (m, 6H). 31 P NMR (162 MHz, Chloroform-d) δ 2.08, 2.20. LCMS: MS m / z = 834.5 [2M+Na]; 405.1 [M-1], t R = 1.33 min; LC system: Thermo Dionex ultimate 3000 UHPLC; column: Phenomenex Kinetex 2.6 μ C18 100 A, 50 x 3 mm; solvents: A: water with 0.1% acetic acid, B: acetonitrile with 0.1% acetic acid; gradient: 0 min - 0.3 min 5% B, 0.3 min - 1.5 min 5% - 100% B, 1.5 min - 2 min 100% B, 2 min - 2.2 min 100% - 5% B, at a rate of 2 mL / min. HPLC: t R = 3.60 min; HPLC system: Agilent 1100 series; column: Phenomenex Gemini 5 μ C18 110 A, 50 x 4.6 mm; solvents: A: water with 0.1% TFA, B: acetonitrile with 0.1% TFA; gradient: 2% - 98% B over 5 min, at a rate of 2 mL / min.

[0708] Intermediate 74. ((S)-(4-nitrophenoxy)(phenoxy) phosphoryl)-L-alanine cyclohexyl ester

[0709]

[0710] Phosphorus dichloride, 4-nitrobenzene (512 mg, 2 mmol) was mixed with 10 mL of dry dichloromethane and stirred in an ice bath under atmospheric nitrogen. L-alanine isopropyl ester hydrochloride (335 mg, 2 mmol) was dissolved in dry dichloromethane (3 mL) and added dropwise to the reaction. The reaction mixture was stirred for 30 min. Triethylamine (927 μL, 6.6 mmol) was dissolved in dry dichloromethane (1 mL) and added dropwise to the reaction mixture. The reaction mixture was stirred for 60 min. 2-Methoxyethanol (158 μL, 2 mmol) was added in one portion and the reaction mixture was stirred for 16 hours. The reaction mixture was diluted with dichloromethane (15 mL) and washed with water (3 x 10 mL). The organic extract was dried over anhydrous sodium sulfate and concentrated under reduced pr...

Claims

1. A compound of Formula (II): or a pharmaceutically acceptable salt thereof, wherein: Base is (C) phenyl, wherein (A) -OH; R 1A and R 2A each independently is: (A) optionally substituted by 1 to 3 R 1B substituted C 1-12 alkyl, (B) 3- to 6-membered heterocyclyl having 1 to 3 heteroatoms independently selected from N, O, and S, wherein the 3- to 6-membered heterocyclyl is optionally substituted with 1 to 3 R 1C substituents, or m is 0, 1, 2, 3, 4, or 5; and each R is independently -OH, -NH2, C 1B is independently -OH, -NH2, C 1-6 alkoxy, methoxyethoxy, or a 3- to 6-membered heterocyclyl having 1 to 3 heteroatoms independently selected from N, O, and S, and Each R 1C Independently for C 1-3 alkyl; R 3 -N(H)R 3A or -N=C(R 3B )(R 3C ); R 3A is H, -CH2OP(O)(OH)2, or -C(O)R 3D wherein R 3D C, optionally substituted with one methoxy group 1-6 Alkyl, or optionally C 1-3 Alkyl-substituted 3- to 6-membered heterocyclic groups having 1 to 3 heteroatoms independently selected from N, O, and S; R 3B R is H or C 1-3 alkyl; R 3C is -N(R 3C1 )(R 3C2 ); R 3C1 and R 3C2 each independently H or C 1-6 alkyl; or R 3C1 and R 3C2 together with the atoms to which they are attached form a 3- to 6-membered heterocyclyl having 1 to 3 heteroatoms independently selected from N, O, and S; and 1-6 alkyl; and R is H, halo, C1-C6alkyl, C1-C6haloalkyl, C3-C6cycloalkyl, C1-C6alkoxy, C1-C6haloalkoxy, C1-C6alkylthio, C1-C6haloalkylth R 4A is O or S; and R 4B and R 4C each independently is 14. The compound of claim 1, or a pharmaceutically acceptable salt thereof, represented by Formula (IIa): (B) -OR 4B1 wherein R 4B1 C 4B2 1 to 3 R 1-6 alkyl, or C 6-12 aryl, wherein each R is independently C 4B2 group is independently C 1-6 alkoxy, -S-R 4B3 or -S(O)2-R 4B3 and each R is independently C 4B3 group is independently C 1-6 alkyl; (C) wherein 15. The compound of claim 1, or a pharmaceutically acceptable salt thereof, represented by Formula (IIb): Each R 4D Independently, it can be optionally divided into 1 to 3 Rs. 4D1 C with substituent group 1-3 Alkyl groups, optionally with 1 to 3 R groups 4D2 C with substituent group 1-3 alkoxy group, or -C(O)N(R) 4D3 )2, of which each R is independently -NH2or -C(O)OR 4D1 group is independently -NH2or -C(O)OR 4D3 , Each R 4D2 Independently for C 1-3 alkoxy, and Each R 4D3 Independently for C 1-3 alkyl; (D) wherein R 4E1 and R 4E2 each independently H or C 1-6 alkyl, R 4F1 and R 4F2 each independently is H or C 1-6 alkyl, or R 4F1 and R 4F2 together are oxo, R 4G To be arbitrarily selected by 1 to 3 R 4G1 Replacement C 1-12 Alkyl groups, optionally with 1 to 3 R groups 4G2 Replacement C 3-7 cycloalkyl, optionally with 1 to 3 R 4G3 Substituted 3- to 8-membered heterocyclic groups having 1 to 3 heteroatoms selected from N, O, and S, or -C(O)R 4G4 , each R is independently -OH, C 4G1 is independently -OH, C 1-6 alkyl, C 1-3 alkoxy, -(CH2OCH2) 1-5 -CH3, -N(R 4G8 )2, -OP(O)(OH)2, C 4G9 cycloalkyl, 3- to 6-membered heterocyclyl having 1 to 3 heteroatoms independently selected from N, O, and S, optionally substituted with 1 to 3 R 3-7 cycloalkyl, 3- to 6-membered heterocyclyl having 1 to 3 heteroatoms independently selected from N, O, and S, optionally substituted with 1 to 3 R 4G10 cycloalkyl, 3- to 6-membered heterocyclyl having 1 to 3 heteroatoms independently selected from N, O, and S, optionally substituted with 1 to 3 R each R is independently C 4G2 is independently C 1-6 alkyl, C 1-3 haloalkyl or -NH2, each R 4G3 is independently halogen or C 1-3 alkyl; each R is independently C 4G4 independently C 1-12 alkyl, each R is independently C 4G8 independently C 1-6 alkyl, each R is independently C 4G9 independently C 1-3 haloalkyl or -NH2, and Each R 4G10 Independently for C 1-3 Halogenated alkyl; or (E) - (OP (O) (OH)) 1-2 -OH; and R 5A and R 5B each is C 1-6 alkyl substituted by -OP(O)(OH)2.

2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2A are C 1B alkyl optionally substituted with 1 to 3 R 1-12 .

3. The compound of claim 2, or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2A are each selected from methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, and i-pentyl, each optionally substituted with 1 to 3 R 1B .

4. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R 4B and one of R 4C is: and R 4B and the other of R 4C is:

5. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R 4F1 and R 4F2 together are oxo.

6. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R 4G is C 4G1 alkyl optionally substituted with 1 to 3 R 1-12 alkyl.

7. The compound of claim 6, or a pharmaceutically acceptable salt thereof, wherein R 4G is methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, or 2-ethylbutyl, each optionally substituted with 1 to 3 R 4G1 .

8. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R 4G is C 4G2 optionally substituted with 1 to 3 R 3-7 cycloalkyl.

9. The compound of claim 8, or a pharmaceutically acceptable salt thereof, wherein R 4G is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, each optionally substituted with 1 to 3 R 4G2 .

10. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R 4G is 3- to 6-membered heterocyclyl having 1 to 3 heteroatoms selected from N, O, and S optionally substituted with 1 to 3 R 4G3 .

11. The compound of claim 10, or a pharmaceutically acceptable salt thereof, wherein R 4G is 4-6 membered heterocyclyl having 1 to 2 heteroatoms selected from N and O optionally substituted with 1 to 3 R 4G3 .

12. The compound of claim 10, or a pharmaceutically acceptable salt thereof, wherein R 4G is oxetanyl, pyrrolidinyl, piperidinyl, tetrahydrofuranyl, or tetrahydropyranyl, each optionally substituted with 1 to 3 R 4G3 .

13. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R 5A and R 5B are each -CH2OP(O)(OH)2.

16. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein: wherein R 5A is -CH2OP(O)(OH)2. ethyl optionally substituted with methoxy, wherein R 5B is -CH2OP(O)(OH)2. n-propyl, R 1A and R 2A each independently is methyl optionally substituted with methoxy, methoxyethoxy, or morpholinyl, isopropyl, n-butyl, isobutyl optionally substituted with -OH or -NH2, tert-butyl, isopentyl, oxetanyl, tetrahydropyranyl, piperidinyl optionally substituted with methyl, or phenyl; ethyl optionally substituted with methoxy, isopropyl, or R 3 -N(H)R 3A or -N=C(R 3B )(R 3C ); R 3A is H, -C(H)2OP(O)(OH)2, or -C(O)R 3D ; R 3D is methyl, piperidinyl optionally substituted with methyl; (A) -OH; m is 0, 1, or 2; and R 3B is H or methyl; R 3C -N(R 3C1 )(R 3C2 ); R 3C1 and R 3C2 are independently H or methyl; or R 3C1 and R 3C2 together with the atoms on which they are attached form piperazinyl optionally substituted with methyl; R 4A is O or S; and R 4B and R 4C each independently is n-propyl optionally substituted with methoxy or morpholinyl, (B) -O-C optionally substituted with methoxy, methylthio, or methylsulfonyl 1-6 alkyl; (C) wherein isopropyl, Each R 4D Independently, for optional use, by 1 R 4D1 C with substituent group 1-3 Alkyl groups, C groups optionally substituted with methoxy groups 1-3 alkoxy group, or -C(O)N(R) 4D3 )2, of which each R 4D1 group is independently -NH2or -C(O)OR 4D3 , and each R 4D3 is independently C 1-3 alkyl; or (D) wherein R 4E1 and R 4E2 each independently H or methyl; R 4F1 and R 4F2 are each independently H or methyl, or R 4F1 and R 4F2 together are oxo; R 4G For optional use by R 4G1 Substituted methyl groups, optionally substituted by morpholinyl or -N(C 1-3 ethyl optionally substituted by morpholinyl or -N(C ethyl optionally substituted by morpholinyl or -N(C isobutyl optionally substituted with -OH or -OP(O)(OH)2, cyclopropyl, optionally substituted by C 1-3 alkyl-substituted n-butyl, cyclobutyl, cyclopentyl, oxetanyl, pyrrolidinyl optionally substituted with 1 to 3 methyl groups, optionally substituted by 1 to 3 groups independently selected from -NH2, C 1-6 alkyl or C 1-3 cycloalkyl, cycloalkyl substituted by 1 to 3 groups independently selected from -NH2, C tetrahydrofuranyl, tetrahydropyranyl, or optionally substituted by halogen or C 1-3 alkyl substituted piperidinyl, cyclobutyl, oxetanyl, -C(O)C 1-6 alkyl; and R 4G1 is cyclopropyl, tetrahydropyranyl, or optionally substituted by -NH2or C 1-3 haloalkyl-substituted cyclohexyl, phenyl. optionally substituted by C 1-3 haloalkyl-substituted piperidinyl, 17. The compound of claim 1, or a pharmaceutically acceptable salt thereof, represented by Formula (IIc):

18. The compound of claim 1, or a pharmaceutically acceptable salt thereof, represented by Formula (IId):

19. The compound of claim 1, or a pharmaceutically acceptable salt thereof, represented by Formula (IIe):

20. The compound of claim 1, or a pharmaceutically acceptable salt thereof, represented by Formula (IIf):

21. The compound of claim 1, or a pharmaceutically acceptable salt thereof, represented by Formula (IIg): wherein m is 0 or 1.

22. The compound of claim 1, or a pharmaceutically acceptable salt thereof, represented by Formula (IIh):

23. The compound of claim 1, or a pharmaceutically acceptable salt thereof, represented by Formula (IIi):

24. The compound of claim 1, or a pharmaceutically acceptable salt thereof, represented by Formula (IIj):

25. The compound of claim 1, or a pharmaceutically acceptable salt thereof, represented by Formula (IIk):

26. The compound of claim 1, or a pharmaceutically acceptable salt thereof, represented by Formula (IIm):

27. The compound of claim 1, or a pharmaceutically acceptable salt thereof, represented by Formula (IIn):

32. The compound of claim 1, or a pharmaceutically acceptable salt thereof, selected from:

33. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is:

28. The compound or pharmaceutically acceptable salt thereof of any one of claims 1 to 27, wherein R 1A and R 2A are each methyl, ethyl, or isopropyl.

29. The compound of claim 28, or a pharmaceutically acceptable salt thereof, wherein R 1A and R 2A are each isopropyl.

30. The compound or pharmaceutically acceptable salt thereof of any one of claims 1 to 27, wherein R 4G is methyl, ethyl, n-propyl, i-propyl, n-butyl, t-butyl, or 2-ethylbutyl.

31. The compound or pharmaceutically acceptable salt thereof of any one of claims 1 to 27, wherein R 4G is methyl. ​ ​ 34. The compound or pharmaceutically acceptable salt thereof of claim 33, wherein the compound is:

35. The compound or pharmaceutically acceptable salt thereof of claim 33, wherein the compound is:

36. The compound or pharmaceutically acceptable salt thereof of claim 33, wherein the compound is:

37. The compound or pharmaceutically acceptable salt thereof of claim 33, wherein the compound is:

38. The compound or pharmaceutically acceptable salt thereof of claim 33, wherein the compound is:

39. The compound or pharmaceutically acceptable salt thereof of claim 33, wherein the compound is:

40. The compound or pharmaceutically acceptable salt thereof of claim 33, wherein the compound is:

41. The compound or pharmaceutically acceptable salt thereof of claim 33, wherein the compound is:

42. The compound or pharmaceutically acceptable salt thereof of claim 33, wherein the compound is:

43. A pharmaceutical formulation comprising a pharmaceutically effective amount of a compound or pharmaceutically acceptable salt thereof of any one of claims 1 to 42, and a pharmaceutically acceptable carrier or excipient.

44. A method of making a medicament for treating a respiratory syncytial virus infection in a human in need thereof, comprising, using one of the following compounds or a pharmaceutically acceptable salt thereof:

45. A method of making a medicament for treating a human metapneumovirus infection in a human in need thereof, comprising, using one of the following compounds or a pharmaceutically acceptable salt thereof:

46. A method of making a medicament for treating a human rhinovirus infection in a human in need thereof, comprising, using one of the following compounds or a pharmaceutically acceptable salt thereof:

47. A method of making a medicament for treating a dengue virus infection in a human in need thereof, comprising, using one of the following compounds or a pharmaceutically acceptable salt thereof:

48. A method of making a medicament for treating an HCV infection in a human in need thereof, comprising combining an effective amount of a compound of fonnula (I), or a pharmaceutically acceptable salt thereof, with a pharmaceutically acceptable carrier. using one of the following compounds or a pharmaceutically acceptable salt thereof:

49. Use of a compound for the manufacture of a medicament for the treatment of a respiratory syncytial virus infection in a human, wherein the compound is selected from: or a pharmaceutically acceptable salt thereof.

50. Use of a compound for the manufacture of a medicament for the treatment of a human metapneumovirus infection in a human, wherein the compound is: or a pharmaceutically acceptable salt thereof.

51. Use of a compound for the manufacture of a medicament for the treatment of a human rhinovirus infection in a human, wherein the compound is selected from: or a pharmaceutically acceptable salt thereof.

52. Use of a compound for the manufacture of a medicament for the treatment of a dengue virus infection in a human, wherein the compound is selected from: or a pharmaceutically acceptable salt thereof.

53. Use of a compound for the manufacture of a medicament for the treatment of an HCV infection in a human, wherein the compound is selected from: or a pharmaceutically acceptable salt thereof.

54. A compound, or a pharmaceutically acceptable salt thereof, characterized in that a compound or pharmaceutically acceptable salt thereof for use in the treatment of a respiratory syncytial virus infection in a human in need thereof, wherein the compound is selected from: or a pharmaceutically acceptable salt thereof.

55. A compound, or a pharmaceutically acceptable salt thereof, characterized in that a compound or pharmaceutically acceptable salt thereof for use in the treatment of a human metapneumovirus infection, wherein the compound is selected from: or a pharmaceutically acceptable salt thereof.

56. A compound, or a pharmaceutically acceptable salt thereof, characterized in that a compound or pharmaceutically acceptable salt thereof for use in the treatment of a human rhinovirus infection, wherein the compound is selected from: or a pharmaceutically acceptable salt thereof.

57. A compound, or a pharmaceutically acceptable salt thereof, characterized in that a compound or pharmaceutically acceptable salt thereof for use in the treatment of a dengue virus infection in a human in need thereof, wherein the compound is selected from: or a pharmaceutically acceptable salt thereof.

58. A compound, or a pharmaceutically acceptable salt thereof, characterized in that the compound is selected from the group consisting of: or a pharmaceutically acceptable salt thereof.

59. A method of manufacturing a medicament for the treatment or prevention of a worsening of chronic obstructive pulmonary disease caused by respiratory syncytial virus, the method comprising combining a therapeutically effective amount of an anti-respiratory syncytial virus antibody with a pharmaceutically acceptable carrier. using one of the following compounds or a pharmaceutically acceptable salt thereof:

60. Use of a compound or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment or prevention of exacerbations of chronic obstructive pulmonary disease caused by respiratory syncytial virus in a human, wherein the compound is selected from the group consisting of: or a pharmaceutically acceptable salt thereof.

61. A compound, or a pharmaceutically acceptable salt thereof, characterized in that the compound is selected from the group consisting of: or a pharmaceutically acceptable salt thereof. the compound is selected from the group consisting of: or a pharmaceutically acceptable salt thereof.

Images