Compounds useful in HIV therapy

Abstract

The present invention relates to compounds of formula (I), their salts, pharmaceutical compositions thereof, and methods of treating or preventing HIV in a subject.

Description

[Technical Field]

[0001] The present invention relates to compounds, pharmaceutical compositions and methods of use thereof. In particular, such methods of use include, for example, methods of treating HIV and methods of preventing HIV. [Background technology]

[0002] Human immunodeficiency virus type 1 (HIV-1) infection leads to the development of acquired immunodeficiency disease (AIDS). The number of HIV cases continues to increase, and an estimated 35 million people worldwide are currently living with HIV infection (e.g., http: / / www.sciencedirect.com / science / article / pii / S235230181630087X?via%3Dihub).

[0003] Currently, long-term suppression of viral replication with antiretroviral drugs is the only option for treating HIV-1 infection. In fact, the U.S. Food and Drug Administration has approved 25 drugs across six different inhibitor classes, which have been shown to significantly improve patient survival and quality of life. However, due to multiple issues, including but not limited to, undesirable drug-drug interactions, drug-food interactions, non-adherence to therapy, drug resistance due to mutations in enzyme targets, and inflammation associated with immunological damage caused by HIV infection, additional therapies are still considered necessary.

[0004] Currently, almost all HIV-positive patients are treated with a combination of antiretroviral drugs, a treatment regimen called highly active antiretroviral therapy ("HAART"). However, HAART therapy is frequently complicated because different drug combinations must be administered to patients almost daily to avoid the rapid emergence of drug-resistant HIV-1 variants. Despite the positive impact of HAART on patient survival, drug resistance can still occur, and survival and quality of life have not normalized compared with uninfected individuals [Lohse Ann Intern Med 2007 146;87-95]. In fact, the morbidity and mortality of several non-AIDS conditions, such as cardiovascular disease, frailty, and neurocognitive disorders, are increased in HIV-infected subjects who are suppressed by HAART [Deeks Annu Rev Med 2011;62:141-155]. The increased incidence of non-AIDS morbidity and mortality occurs in association with and may be caused by elevated systemic inflammation associated with immunological damage caused by HIV infection [Hunt J Infect Dis 2014][Byakagwa J Infect Dis 2014][Tenorio J Infect Dis 2014].

[0005] Current antiretroviral therapy (ART) can effectively suppress HIV replication and improve the health of HIV-infected individuals, but it is believed that it cannot completely eliminate the HIV viral reservoir within an individual. The HIV genome remains latent within most immune cells of infected individuals and can reactivate at any time. Consequently, viral replication typically resumes within a few weeks after ART is discontinued. In a minority of individuals, the size of this viral reservoir is significantly reduced, delaying the rebound of viral replication upon ART cessation [Henrich TJ J Infect Dis 2013] [Henrich TJ Ann Intern Med 2014]. In one case, the viral reservoir was eliminated during treatment for leukemia, and no viral rebound was observed during several years of follow-up [Hutter GN Engl J Med 2009]. These examples suggest the concept that the viral reservoir may be reduced or eliminated, potentially leading to viral remission or cure. As a result, methods have been pursued to eliminate the viral reservoir by direct molecular means, such as by excising the viral genome using the CRISPR / Cas9 system, or to induce reactivation of the latent reservoir during ART, thereby eliminating latent cells. Induction of the latent reservoir typically involves direct death of latently infected cells or the immune system killing the induced cells after the virus becomes visible. Because this occurs during ART, it is believed that the generated viral genomes do not infect new cells, thereby reducing the size of the reservoir.

[0006] HAART therapy is frequently complex because different drug combinations must be administered to patients almost daily to avoid the rapid emergence of drug-resistant HIV-1 variants. Despite the positive impact of HAART on patient survival, drug resistance can still occur.

[0007] Current guidelines recommend that treatment include three fully effective drugs. See, for example, https: / / aidsinfo.nih.gov / guidelines. Furthermore, two-drug combinations can be used as treatment regimens. First-line therapy typically combines two to three drugs targeting the viral enzymes reverse transcriptase and integrase. It is believed that sustained success in treating HIV-1-infected patients with antiretroviral drugs requires the continued development of new, improved drugs that are effective against HIV strains that have developed resistance to approved drugs. For example, individuals on regimens containing 3TC / FTC (lamivudine / emtricitabine) may exhibit the M184V mutation, which reduces susceptibility to these drugs by more than 100-fold. See, for example, https: / / hivdb.stanford.edu / dr-summary / resistance-notes / NRTIs

[0008] Another approach to preventing mutations is to increase patient adherence to drug regimens. One way to achieve this is to reduce the frequency of administration. For parenteral administration, it is considered advantageous to have a highly lipophilic drug substance to reduce solubility and limit the release rate in interstitial fluid. However, the hydrophilicity of most nucleoside reverse transcriptase inhibitors may limit their use as long-acting parenteral agents.

[0009] There remains a need for compounds that can address the above shortcomings. Summary of the Invention

[0010] In one aspect, the present invention provides a compound of formula (I): [ka] [In the formula, R 1 is (C1-C 25 ) alkyl, (C1-C25 ) alkylene aryl, (C1-C 20 )Alkylene-CO2R 5 and aryl; R 2 is (C1-C 10 ) alkyl, (C1-C 10 ) selected from the group consisting of alkylene, aryl, and aryl; R 3 is (C1-C 25 ) alkyl, (C1-C 25 ) selected from the group consisting of alkylene, aryl, and aryl; R 4 is (C1-C 25 ) alkyl, (C1-C 25 ) alkylene aryl, (C1-C 20 )Alkylene-CO2R 5 and aryl; X is a bond, NR 6 or O, R 5 is (C1-C 25 ) alkyl, (C1-C 25 ) selected from the group consisting of alkylene, aryl, and aryl; R 6 is H, (C1-C 25 ) alkyl, (C1-C 25 ) alkylene aryl, (C1-C 20 )Alkylene-CO2R 5 and aryl; R 1 , R 2 , R 3 , R 4 , R 5 and R 6 Each of the groups independently represents one or more (C1-C 14 ) optionally substituted with alkyl, Cl, F, oxo or (C1-C6)alkoxy. or a pharmaceutically acceptable salt thereof.

[0011] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and an excipient.

[0012] In another aspect, the present invention provides a combination comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and one or more agents active against HIV.

[0013] In another aspect, the present invention provides a method of treating HIV infection in a patient, comprising administering to the patient a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0014] In another aspect, the present invention provides a method for preventing HIV infection in a subject at risk of developing HIV infection, the method comprising administering to the subject a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0015] In another aspect, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in therapy.

[0016] In another aspect, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of HIV infection.

[0017] In another aspect, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the prevention of HIV infection.

[0018] In another aspect, there is provided the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of HIV infection.

[0019] In another aspect, there is provided the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the prevention of HIV infection.

[0020] In another aspect, the present invention provides a method of treating HIV infection in a patient, which method comprises administering to the patient a combination comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and one or more agents effective against HIV.

[0021] In another aspect, the present invention provides a method for preventing HIV infection in a subject at risk of developing HIV infection, the method comprising administering to the subject a combination comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and one or more agents effective against HIV.

[0022] In another aspect, there is provided a combination comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and one or more agents active against HIV for use in therapy.

[0023] In another aspect, there is provided a combination comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and one or more agents active against HIV for use in the treatment of HIV infection.

[0024] In another aspect, there is provided a combination comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and one or more agents active against HIV for use in the prevention of HIV infection.

[0025] In another aspect, there is provided the use of a combination comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and one or more agents active against HIV in the manufacture of a medicament for the treatment of HIV infection.

[0026] In another aspect, there is provided the use of a combination comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and one or more agents active against HIV in the manufacture of a medicament for the prevention of HIV infection.

[0027] These and other aspects are encompassed by the invention described herein. DETAILED DESCRIPTION OF THE INVENTION

[0028] Throughout this application, reference is made to various embodiments relating to compounds, compositions, and methods. The various embodiments described are intended to provide various illustrative examples and should not be construed as descriptions of alternative species. Rather, it should be noted that the descriptions of various embodiments provided herein may be of overlapping scope. The embodiments discussed herein are merely exemplary and are not meant to limit the scope of the invention.

[0029] It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention. In this specification and the claims that follow, reference will be made to certain terms that shall be defined to have the following meanings, unless otherwise indicated herein by a described embodiment.

[0030] The term "alkyl" refers to a saturated straight or branched chain hydrocarbon group. For illustrative purposes, an example is "(C1-C 25 The term "alkyl" refers to an alkyl group containing 1 to 6 carbon atoms. Exemplary alkyls include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, s-butyl, t-butyl, pentyl, and hexyl, as well as long chain alkyls.

[0031] The term "alkylene" refers to a divalent straight or branched chain hydrocarbon group. Non-limiting examples include (C-C 25 ) alkylene-aryl and (C1-C 20 )Alkylene-CO2R 4 The term is intended, for purposes of illustration, to represent an alkylene moiety containing 1 to 25 or 1 to 20 carbon atoms, having one or more groups, which may be the same or different, as described, on one or more carbon atoms of the alkylene moiety, which may be a straight or branched chain carbon moiety.

[0032] "Alkoxy" refers to a group containing an alkyl moiety, as defined above, attached through an oxygen linking atom. As an example, the term "(C-C)alkoxy" refers to a straight or branched chain hydrocarbon group having from 1 to 4 carbon atoms attached through an oxygen linking atom. In this particular example, exemplary "(C-C)alkoxy" groups include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, s-butoxy, isobutoxy, and t-butoxy.

[0033] The term "aryl" refers to an aryl group having, for example, 6 to 14 carbon atoms (e.g., (C6-C 14 )aryl) represents a monocyclic or fused bicyclic group having at least one aromatic ring according to Hückel's rule. Examples of "aryl" groups are phenyl (C6), naphthyl, indenyl, dihydroindenyl, anthracenyl, and phenanthrenyl.

[0034] The terms "halogen" and "halo" refer to a chloro, fluoro, bromo or iodo substituent.

[0035] The term "oxo" refers to a double-bonded oxygen moiety, for example, when attached directly to a carbon atom to form a carbonyl moiety (C=O).

[0036] The term "pharmaceutically acceptable" refers to compounds, materials, compositions and dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, or other problem or complication commensurate with a reasonable benefit / risk ratio.

[0037] The term "pharmaceutically acceptable salt" refers to a salt that retains the desired biological activity of the subject compound and minimizes undesired toxicological effects. These pharmaceutically acceptable salts can be prepared in situ during the final isolation and purification of the compound, or by separately reacting the purified compound in its free acid or free base form with a suitable base or acid, respectively. Non-limiting examples include sodium, potassium, calcium, magnesium, ammonium, and tetraalkylammonium salts, as well as, if the molecule contains a basic functional group, salts of organic or inorganic acids, such as hydrochloride, hydrobromide, tartrate, mesylate, acetate, maleate, and oxalate salts. Suitable salts include those described in P. Heinrich Stahl, Camille G. Wermuth (Eds.), Handbook of Pharmaceutical Salts Properties, Selection, and Use; 2002. For example, pharmaceutically acceptable salts of the present invention can be synthesized from a parent compound containing a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of an appropriate base or acid in water or an organic solvent, or a mixture of the two. Non-aqueous media, such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile, are generally preferred as the solvent. The salts may be precipitated from the solution and collected by filtration, or recovered by evaporation of the solvent. The degree of ionization in the salts can vary from completely ionized to almost non-ionized. A list of suitable salts can be found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418 (the disclosure of which is incorporated herein by reference solely for the list of suitable salts).

[0038] The term "preventing" disease or "preventing" disease in a patient refers to the prophylactic administration of a drug to substantially reduce the likelihood or severity of a disorder or its biological manifestations, or to delay the onset of such a disorder or its biological manifestations.

[0039] The term "treatment" refers to alleviating a particular condition, eliminating or alleviating one or more symptoms of a condition, slowing or eliminating the progression of a condition, and delaying the recurrence of a condition in a previously affected or diagnosed patient or subject.

[0040] The term "heteroatom" means nitrogen, oxygen, or sulfur, including any oxidized form of phosphorus, any oxidized form of nitrogen, for example, N(O){N + -O -}, including any oxidized form of sulfur, e.g., S(O) and S(O)2, and the quaternized form of any basic nitrogen.

[0041] The term "patient" or "subject" refers to a mammal, and includes human and non-human mammals. Most preferably, "patient" is taken to refer to a human.

[0042] In one embodiment, the compound of formula (I): [ka] [In the formula, R 1 is (C1-C 25 ) alkyl, (C1-C 25 ) alkylene aryl, (C1-C 20 )Alkylene-CO2R 5 and aryl; R 2 is (C1-C 10 ) alkyl, (C1-C 10 ) selected from the group consisting of alkylene, aryl, and aryl; R 3 is (C1-C 25 ) alkyl, (C1-C 25) selected from the group consisting of alkylene, aryl, and aryl; R 4 is (C1-C 25 ) alkyl, (C1-C 25 ) alkylene aryl, (C1-C 20 )Alkylene-CO2R 5 and aryl; X is a single bond, NR 6 or O, R 5 is (C1-C 25 ) alkyl, (C1-C 25 ) selected from the group consisting of alkylaryl and aryl; R 6 is H, (C1-C 25 ) alkyl, (C1-C 25 ) alkylaryl, (C1-C 20 )Alkyl-CO2R 5 and aryl; R 1 , R 2 , R 3 , R 4 , R 5 and R 6 Each of the groups independently represents one or more (C1-C 14 ) optionally substituted with alkyl, Cl, F, oxo or (C1-C6)alkoxy. or a pharmaceutically acceptable salt thereof.

[0043] In one embodiment, R 1 (C6-C 14 ) aryl. More preferably, R 1 is a C6 aryl.

[0044] In one embodiment, R 1 (C1-C 20 )Alkylene-CO2R 5 More preferably, R 1 (C1-C 20 )Alkylene-CO2R 5 Selected from R 5 (C1-C25 ) alkyl.

[0045] In one embodiment, R 2 (C1-C 10 ) Alkylene-(C6-C 14 ) aryl. More preferably, R 2 is C1 alkylene C6 aryl.

[0046] In one embodiment, R 2 (C1-C 10 ) alkyl. In another embodiment, R 2 is (C1-C5) alkyl. More preferably, R 2 is a C1 alkyl.

[0047] In one embodiment, R 3 (C1-C 25 ) alkyl, for example (C5-C 25 ) alkyl. In another embodiment, R 3 is (C1-C4) alkyl. In another embodiment, R 3 (C 10 -C 15 ) alkyl. In another embodiment, R 3 (C 15 -C 25 ) alkyl.

[0048] In one embodiment, R 4 (C1-C 25 ) alkyl. In another embodiment, R 4 (C5-C 10 ) alkyl. In another embodiment, R 4 (C 15 -C 25 ) alkyl.

[0049] In one embodiment, X is a bond.

[0050] In one embodiment, X is "O".

[0051] In one embodiment, X is NR 6 is.

[0052] In one embodiment, R 1 is a C6 aryl, and R 2 is C1 alkyl or C1 alkylene C6 aryl, and R 3 (C1-C 25 ) alkyl, and R 4 (C1-C 25 alkyl), and X is a bond.

[0053] In one embodiment, R 1 is a C6 aryl, and R 2 is C1 alkyl or C1 alkylene C6 aryl, and R 3 (C1-C 25 ) alkyl, and R 4 (C1-C 25 alkyl) and X is O.

[0054] In one embodiment, R 1 is a C6 aryl, and R 2 is C1 alkyl or C1 alkylene C6 aryl, and R 3 (C1-C 25 ) alkyl, and R 4 (C1-C 25 alkyl), and X is NR 6 is.

[0055] In one embodiment, R 1 (C1-C 25 )Alkylene-CO2R 5 and R 2 is C1 alkyl or C1 alkylene C6 aryl, and R 3 (C1-C 25 ) alkyl, and R 4 (C1-C 25 alkyl), and X is a bond.

[0056] In one embodiment, R 1 (C1-C 25 )Alkylene-CO2R5 and R 2 is C1 alkyl or C1 alkylene C6 aryl, and R 3 (C1-C 25 ) alkyl, and R 4 (C1-C 25 alkyl) and X is O.

[0057] In one embodiment, R 1 (C1-C 25 )Alkylene-CO2R 5 and R 2 is C1 alkyl or C1 alkylene C6 aryl, and R 3 (C1-C 25 ) alkyl, and R 4 (C1-C 25 alkyl), and X is NR 6 is.

[0058] In one embodiment, R 2 (C1-C 10 ) alkyl.

[0059] In one embodiment, R 2 is (C1-C5) alkyl.

[0060] In addition to the above embodiments, in further embodiments, the compounds of the present invention may be substituted with one or more of the substituents described below. For example, in one embodiment, R 1 , R 2 , R 3 , R 4 , R 5 and R 6 each of which is optionally independently substituted with one or more (C1-C6) alkyl, Cl, F, oxo, or (C1-C6) alkoxy. Preferably, as an example, each of the aryl groups is optionally substituted with one or more substituents selected from (C1-C5) alkyl, Cl, F, oxo, or (C1-C6) alkoxy.

[0061] As an example, in one embodiment, R2 is C1 alkylene C6 aryl, where C6 aryl is one or more (C1-C 14 ) alkyl, Cl or F. More specifically, C6 aryl may be substituted with one or more F, for example, two F. As a preferred example: C6 aryl is [ka] is.

[0062] In another aspect of the present invention, the present invention can encompass a variety of distinct compounds. By way of example, such particular compounds can be selected from the group consisting of Table 1.

[0063] [Table 1] JPEG2024507550000005.jpg248166 JPEG2024507550000006.jpg239166 JPEG2024507550000007.jpg216166 JPEG2024507550000008.jpg249164 JPEG2024507550000009.jpg237163 JPEG2024507550000010.jpg237162 JPEG2024507550000011.jpg238161 JPEG2024507550000012.jpg240160 JPEG2024507550000013.jpg222160 JPEG2024507550000014.jpg231160 JPEG2024507550000015.jpg232160 JPEG2024507550000016.jpg234157 JPEG2024507550000017.jpg245158 JPEG2024507550000018.jpg241158 JPEG2024507550000019.jpg248159 JPEG2024507550000020.jpg234159 JPEG2024507550000021.jpg223158 JPEG2024507550000022.jpg239157 JPEG2024507550000023.jpg242157 JPEG2024507550000024.jpg225157 JPEG2024507550000025.jpg37158

[0064] In one embodiment, the present invention encompasses the individual compounds listed in Table 1 above, or pharmaceutically acceptable salts thereof.

[0065] Also within the scope of the present invention, in various embodiments, are prodrugs of any of the compounds of formula (I) described herein.

[0066] According to one embodiment of the present invention, there is provided a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient. In a further embodiment, the compound is in amorphous form. In a further embodiment, the compound is in crystalline form. In a further embodiment, the pharmaceutical composition is in tablet form. In a further embodiment, the pharmaceutical composition is in parenteral form. In a further embodiment, the compound is present as a spray-dried dispersion.

[0067] According to one embodiment of the present invention, there is provided a method of treating HIV infection in a subject, comprising administering to the subject a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0068] According to one embodiment of the present invention, there is provided a method of treating HIV infection in a subject, the method comprising administering to the subject a pharmaceutical composition described herein.

[0069] According to one embodiment of the present invention, there is provided a method for preventing HIV infection in a subject at risk of developing HIV infection, the method comprising administering to the subject a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0070] According to one embodiment of the present invention there is provided the use of a compound of formula (I) in the manufacture of a medicament for the treatment of HIV infection.

[0071] According to one embodiment of the present invention there is provided the use of a compound of formula (I) in the manufacture of a medicament for the prevention of HIV infection.

[0072] According to one embodiment of the present invention there is provided a compound of formula (I) for use in the treatment of HIV infection.

[0073] According to one embodiment of the present invention there is provided a compound of formula (I) for use in the prevention of HIV infection.

[0074] According to one embodiment of the present invention, there is provided a method for preventing HIV infection in a subject at risk of developing HIV infection, the method comprising administering to the subject a pharmaceutical composition described herein.

[0075] Furthermore, the compounds of the present invention may be in particular geometric or stereoisomeric forms. The present invention contemplates all such compounds as being within the scope of the present invention, e.g., cis- and trans-isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, racemic mixtures thereof, and other mixtures thereof, e.g., enantiomerically or diastereomerically enriched mixtures. Additional asymmetric carbon atoms may be present in substituents, e.g., alkyl groups. All such isomers and mixtures thereof are intended to be included in the present invention.

[0076] Optically active (R)- and (S)-isomers and d- and l-isomers can be prepared using chiral synthons or chiral reagents or resolved using conventional techniques. For example, if a specific enantiomer of a compound of the present invention is desired, it can be prepared by asymmetric synthesis or by derivatization with a chiral auxiliary. In the latter case, the resulting diastereomeric mixture is separated, and the auxiliary is cleaved to provide the pure desired enantiomer. Alternatively, if the molecule contains a basic functional group, such as an amino group, or an acidic functional group, such as a carboxyl group, diastereomeric salts can be formed using an appropriate optically active acid or base, and the diastereomers thus formed can then be resolved by fractional crystallization or chromatographic means known in the art, followed by recovery of the pure enantiomer. Furthermore, separation of enantiomers and diastereomers is often achieved using chromatography using a chiral stationary phase, optionally in combination with chemical derivatization (e.g., formation of carbamates from amines).

[0077] In another embodiment of the invention there is provided a compound of formula (I), wherein the compound or a salt of the compound is used in the manufacture of a medicament for use in the treatment of HIV infection in humans.

[0078] In another embodiment of the invention there is provided a compound of formula (I) wherein the compound or a salt of the compound is used in the manufacture of a medicament for use in the prevention of HIV infection in humans.

[0079] In one embodiment, the pharmaceutical formulation comprising a compound of formula (I) or a salt thereof is a formulation adapted for parenteral administration. In another embodiment, the formulation is a long-acting parenteral formulation. In a further embodiment, the formulation is a nanoparticle formulation.

[0080] The compounds of the present invention and their salts, solvates, or other pharmaceutically acceptable derivatives may be used alone or in combination with other therapeutic agents. Thus, in other embodiments, the method for treating and / or preventing HIV infection in a subject may further comprise, in addition to administering a compound of formula (I), administering one or more additional pharmaceutical agents effective against HIV.

[0081] In such embodiments, the one or more additional agents effective against HIV are zidovudine, didanosine, lamivudine, zalcitabine, abacavir, stavudine, adefovir, adefovir dipivoxilDipivoxil, fozivudine, todoxil, emtricitabine, alovudine, amdoxovir, elvucitabine, nevirapine, delavirdine, efavirenz, loviride, immunocal, oltipraz ipraz, capravirine, lersivirine, GSK2248761, TMC-278, TMC-125, etravirine, saquinavir, ritonavir, indinavir, nelfinavir, amprenavir, fosamprenavir, brecanavir ir), darunavir, atazanavir, tipranavir, parinavir, lasinavir, enfuvirtide, T-20, T-1249, PRO-542, PRO-140, TNX-355, BMS-806, BMS-663068, BMS-626529, 5-Helix, raltegravir, elvitegravir (elvitegravir), dolutegravir, cabotegravir, bictegravir, vicriviroc (Sch-C), Sch-D, TAK779, maraviroc, TAK449, didanosine, tenofovir, lopinavir, and darunavir.

[0082] Thus, the compound of the present invention of formula (I) and any other one or more active pharmaceutical ingredients may be administered together or separately, and when administered separately, they may be administered simultaneously or sequentially in any order. The amounts of the compound of formula (I) of the present invention and the other one or more active pharmaceutical ingredients and the relative timing of administration are selected to achieve the desired combined therapeutic effect. The compound of formula (I) of the present invention and its salts, solvates, or other pharmaceutically acceptable derivatives thereof in combination with other therapeutic agents may be administered in the form of (1) a single pharmaceutical composition containing both compounds; or (2) separate pharmaceutical compositions each containing one of the compounds simultaneously. Alternatively, the combination may be administered separately and sequentially, with one therapeutic agent being administered first and the other being administered second, or vice versa. Such sequential administration may be close in time or distant in time. The amounts of one or more compounds of formula (I) or salts thereof and one or more other active pharmaceutical ingredients and the relative timings of administration will be selected in order to achieve a desired combined therapeutic effect.

[0083] Furthermore, the compounds of the present invention of formula (I) may be used in combination with one or more other drugs that may be useful in the prevention or treatment of HIV. Examples of such drugs include the following: Nucleoside reverse transcriptase inhibitors, such as zidovudine, didanosine, lamivudine, zalcitabine, abacavir, stavudine, adefovir, adefovir dipivoxil, fodivudine, todoxil, emtricitabine, alovudine, amdoxovir, elvucitabine and similar agents; Non-nucleoside reverse transcriptase inhibitors (e.g., drugs with antioxidant activity, such as Immunocal, Oltipraz, etc.), such as nevirapine, delavirdine, efavirenz, loviride, Immunocal, Oltipraz, capravirine, lersivrine, doravirine, GSK2248761, TMC-278, TMC-125, etravirine, and similar drugs; Protease inhibitors, such as saquinavir, ritonavir, indinavir, nelfinavir, amprenavir, fosamprenavir, brecanavir, darunavir, atazanavir, tipranavir, parinavir, lasinavir and similar agents; Entry, adsorption and fusion inhibitors, such as enfuvirtide (T-20), T-1249, PRO-542, PRO-140, TNX-355, BMS-806, BMS-663068 (fostemsavir), BMS-626529 (temsavir), 5-helix and similar agents; Integrase inhibitors, such as raltegravir, elvitegravir, dolutegravir, bictegravir, cabotegravir and similar agents; Maturation inhibitors, such as PA-344 and PA-457, and similar agents; and CXCR4 and / or CCR5 inhibitors, such as vicriviroc (Sch-C), Sch-D, TAK779, maraviroc (UK427,857), TAK449, and agents disclosed in WO02 / 74769, PCT / US03 / 39644, PCT / US03 / 39975, PCT / US03 / 39619, PCT / US03 / 39618, PCT / US03 / 39740 and PCT / US03 / 39732, and similar agents; Capsid inhibitors, such as GS-6207 and similar agents.

[0084] Further examples of compounds of the invention that may be used in combination with one or more agents useful in the prevention or treatment of HIV are found in Table 2.

[0085] [Table 2]

[0086] The scope of the combination of the compound of the present invention with an HIV drug is not limited to those listed above, but includes, in principle, any combination with any pharmaceutical composition useful for the treatment and / or prevention of HIV. As mentioned above, in such combinations, the compound of the present invention and the other HIV drug may be administered separately or together. Furthermore, one drug may be administered before, simultaneously with, or after the administration of one or more other drugs.

[0087] The present invention may be used in combination with one or more agents useful as pharmacological enhancers, with or without additional compounds for the prevention or treatment of HIV. Examples of such pharmacological enhancers (or pharmacokinetic enhancers) include, but are not limited to, ritonavir, GS-9350, and SPI-452.

[0088] Ritonavir is 10-hydroxy-2-methyl-5-(1-methylethyl)-1-[2-(1-methylethyl)-4-thiazolyl]-3,6-dioxo-8,11-bis(phenylmethyl)-2,4,7,12-tetraazatridecan-13-onic acid, 5-thiazolylmethyl ester, [5S-(5S * ,8R * ,10R * ,11R * Ritonavir is an HIV protease inhibitor that is indicated in conjunction with other antiretroviral agents for the treatment of HIV infection. Ritonavir also inhibits P450-mediated drug metabolism and the P-glycoprotein (Pgp) cellular transport system, thereby resulting in increased concentrations of the active compound within the organism.

[0089] GS-9350 is a compound developed by Gilead Sciences (Foster City, California) as a pharmacological enhancer.

[0090] SPI-452 is a compound developed by Sequoia Pharmaceuticals (Gaithersburg, Maryland) as a pharmacological enhancer.

[0091] In one embodiment of the present invention, the compound of formula (I) is used in combination with ritonavir. In one embodiment, the combination is an oral fixed-dose combination. In another embodiment, the compound of formula (I) is formulated as a long-acting parenteral injection, and ritonavir is formulated as an oral composition. In one embodiment, a kit is provided comprising the compound of formula (I) formulated as a long-acting parenteral injection and ritonavir formulated as an oral composition. In another embodiment, the compound of formula (I) is formulated as a long-acting parenteral injection, and ritonavir is formulated as an injectable composition. In one embodiment, a kit is provided comprising the compound of formula (I) formulated as a long-acting parenteral injection and ritonavir formulated as an injectable composition.

[0092] In another embodiment of the present invention, the compound of Formula (I) is used in combination with GS-9350. In one embodiment, the combination is an oral fixed-dose combination. In another embodiment, the compound of Formula (I) is formulated as a long-acting parenteral injection and GS-9350 is formulated as an oral composition. In one embodiment, a kit is provided comprising the compound of Formula (I) formulated as a long-acting parenteral injection and GS-9350 formulated as an oral composition. In another embodiment, the compound of Formula (I) is formulated as a long-acting parenteral injection and GS-9350 is formulated as an injectable composition. In one embodiment, a kit is provided comprising the compound of Formula (I) formulated as a long-acting parenteral injection and GS-9350 formulated as an injectable composition.

[0093] In one embodiment of the present invention, a compound of formula (I) is used in combination with SPI-452. In one embodiment, the combination is an oral fixed-dose combination. In another embodiment, a compound of formula (I) is formulated as a long-acting parenteral injection, and SPI-452 is formulated as an oral composition. In one embodiment, a kit is provided comprising a compound of formula (I) formulated as a long-acting parenteral injection and SPI-452 formulated as an oral composition. In another embodiment, a compound of formula (I) is formulated as a long-acting parenteral injection, and SPI-452 is formulated as an injectable composition. In one embodiment, a kit is provided comprising a compound of formula (I) formulated as a long-acting parenteral injection, and SPI-452 formulated as an injectable composition.

[0094] In one embodiment of the present invention, the compounds of formula (I) are used in combination with compounds found in previously filed PCT / CN2011 / 0013021, which is incorporated herein by reference.

[0095] The other therapeutic agents described above, when used in combination with the chemical entities described herein, may be used, for example, in amounts as set forth in the Physicians' Desk Reference (PDR) or as otherwise determined by one of skill in the art.

[0096] In another embodiment of the present invention, there is provided a method of treating a viral infection in a mammal mediated at least in part by a member of the retrovirus family of viruses, comprising administering a compound of formula (I) to a mammal diagnosed with or at risk of developing said viral infection.

[0097] In another embodiment of the present invention, there is provided a method of treating a viral infection in a mammal mediated at least in part by a member of the retrovirus family of viruses, the method comprising administering a compound of formula (I) to a mammal diagnosed with or at risk of developing said viral infection, wherein said virus is an HIV virus. In some embodiments, the HIV virus is an HIV-1 virus.

[0098] In another embodiment of the present invention, there is provided a method of treating a viral infection in a mammal mediated at least in part by a member of the retrovirus family of viruses, comprising administering to a mammal diagnosed with or at risk of developing said viral infection a compound of formula (I), and further comprising administering a therapeutically effective amount of one or more agents effective against the HIV virus.

[0099] In another embodiment of the present invention, there is provided a method for treating a viral infection in a mammal mediated at least in part by a virus of the retrovirus family of viruses, the method comprising administering a compound of formula (I) to a mammal diagnosed with or at risk of developing said viral infection, and further comprising administering a therapeutically effective amount of one or more agents effective against the HIV virus, wherein the agents effective against the HIV virus are selected from nucleoside reverse transcriptase inhibitors; non-nucleoside reverse transcriptase inhibitors; protease inhibitors; entry, adsorption and fusion inhibitors; integrase inhibitors; maturation inhibitors; capsid inhibitors; CXCR4 inhibitors; and CCR5 inhibitors.

[0100] In another embodiment of the present invention, there is provided a method of preventing a viral infection in a mammal mediated at least in part by a member of the retrovirus family of viruses, comprising administering a compound of formula (I) to a mammal diagnosed with or at risk of developing said viral infection.

[0101] In another embodiment of the present invention, there is provided a method for preventing a viral infection in a mammal mediated at least in part by a virus of the retrovirus family of viruses, the method comprising administering a compound of formula (I) to a mammal diagnosed with or at risk of developing said viral infection, wherein said virus is the HIV virus. In some embodiments, the HIV virus is the HIV-1 virus.

[0102] In another embodiment of the present invention, there is provided a method for preventing a viral infection in a mammal mediated at least in part by a member of the retrovirus family of viruses, comprising administering to a mammal diagnosed with or at risk of developing said viral infection a compound of formula (I), and further comprising administering a therapeutically effective amount of one or more agents effective against the HIV virus.

[0103] In another embodiment of the present invention, there is provided a method for preventing a viral infection in a mammal mediated at least in part by a virus of the retrovirus family of viruses, the method comprising administering a compound of formula (I) to a mammal diagnosed with or at risk of developing said viral infection, and further comprising administering a therapeutically effective amount of one or more agents effective against the HIV virus, wherein the agents effective against the HIV virus are selected from nucleoside reverse transcriptase inhibitors; non-nucleoside reverse transcriptase inhibitors; protease inhibitors; entry, adsorption and fusion inhibitors; integrase inhibitors; maturation inhibitors; capsid inhibitors; CXCR4 inhibitors; and CCR5 inhibitors.

[0104] In a further embodiment, the compound of the invention of formula (I) or a pharmaceutically acceptable salt thereof is selected from the group of compounds set forth in Table 1 above.

[0105] The compounds in Table 1 were synthesized according to the synthetic methods, general schemes and examples described below.

[0106] In another embodiment, there is provided a pharmaceutical composition comprising a pharmaceutically acceptable diluent and a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0107] In certain embodiments, the compound of the invention, or a pharmaceutically acceptable salt thereof, is selected from the compounds set forth in Table 1.

[0108] The compounds of formula (I) of the present invention may be in both unsolvated and solvated forms. The term "solvate" includes a compound of the present invention in combination with one or more pharmaceutically acceptable solvent molecules, such as ethanol. The term "hydrate" is used when the solvent is water. Pharmaceutically acceptable solvates include hydrates and other solvates in which the solvent of crystallization may be isotopically substituted (e.g., DO, d-acetone, d-DMSO).

[0109] Compounds of formula (I) containing one or more asymmetric carbon atoms can exist in two or more stereoisomers. When a compound of formula (I) contains an alkenyl group, an alkenylene group, or a cycloalkyl group, geometric cis / trans (or Z / E) isomers are possible. When a compound contains, for example, a keto group, an oxime group, or an aromatic moiety, tautomeric isomerism ("tautomerism") can occur. A single compound can exhibit two or more types of isomerism.

[0110] Included within the scope of the claimed compounds of the invention are all stereoisomeric, geometric isomeric, and tautomeric forms of the compounds of formula (I), e.g., compounds exhibiting more than one type of isomerism, and mixtures of one or more thereof. Also included are acid or base addition salts in which the counterion is optically active, e.g., D-lactate or L-lysine, or racemic, e.g., DL-tartrate or DL-arginine.

[0111] Cis / trans isomers may be separated by conventional techniques well known to those skilled in the art, such as chromatography and fractional crystallization.

[0112] Conventional techniques for the preparation / isolation of separate enantiomers include chiral synthesis from suitable optically pure precursors, or resolution of the racemate (or racemate of a salt or derivative), e.g., using chiral high performance liquid chromatography (HPLC), supercritical fluid chromatography (SFC).

[0113] Alternatively, the racemate (or racemic precursor) may be reacted with a suitable optically active compound, such as an alcohol, or, if the compound of formula (I) contains an acidic or basic moiety, with an acid or base, such as tartaric acid or 1-phenylethylamine. The resulting diastereomeric mixture may be separated by chromatography and / or fractional crystallization, and one or both of the diastereoisomers may be converted into the corresponding pure enantiomer(s) by means well known to those skilled in the art.

[0114] The chiral compounds of the invention (and their chiral precursors) may be obtained in enantiomerically enriched form by chromatography, typically HPLC or SFC, on a resin having an asymmetric stationary phase, using a mobile phase consisting of a hydrocarbon, typically heptane or hexane, containing 0-50%, typically 2-20%, isopropanol, and 0-5% alkylamine, typically 0.1% diethylamine. Concentration of the eluent yields the enriched mixture.

[0115] Mixtures of stereoisomers may be separated by conventional techniques known to those skilled in the art (see, for example, "Stereochemistry of Organic Compounds" by EL Eliel (Wiley, New York, 1994)).

[0116] The present invention includes all pharmaceutically acceptable isotopically labeled compounds of formula (I) where one or more atoms are replaced by an atom having the same atomic number but an atomic mass or mass number different from the atomic mass or mass number usually found in nature.

[0117] Examples of isotopes that are suitable for compounds of the present invention to contain include isotopes of hydrogen (e.g., 2 H and 3 H), isotopes of carbon (e.g., 11 C. 13 C and 14 C), isotopes of chlorine (e.g., 36 Cl), isotopes of fluorine (e.g., 18 F), isotopes of iodine (e.g., 123 I and 125 I), isotopes of nitrogen (e.g., 13 N and 15 N), isotopes of oxygen (e.g., 15 O. 17 O and 18 O), isotopes of phosphorus (e.g., 32 P) and sulfur isotopes (e.g., 35 S) are mentioned.

[0118] Certain isotopically labeled compounds of formula (I), for example, compounds incorporating a radioactive isotope, are useful in drug and / or substrate tissue distribution studies. 3 H) and carbon-14 (i.e., 14 C) is particularly useful for this purpose given its ease of incorporation and ready means of detection. Heavy isotopes, such as deuterium (i.e., 2 H) may confer certain therapeutic benefits resulting from greater metabolic stability, such as increased half-life in vivo or reduced dosage requirements, and therefore may be preferable in some situations.

[0119] Isotopically labeled compounds of formula (I) may generally be prepared by conventional techniques known to those skilled in the art, or by processes analogous to those described herein, substituting an appropriate isotopically labeled reagent for the previously used non-labeled reagent.

[0120] The compounds of the present invention may be administered as prodrugs. Thus, certain derivatives of the compound of formula (I), which may have little or no pharmacological activity themselves, can be converted to the compound of formula (I) as a "prodrug" when administered to or on the body. An example of a compound that may be included in such a prodrug is 4'-ethylnyl-2-fluoro-2'-deoxyadenosine (EFdA), described, for example, in U.S. Pat. No. 7,339,053. The compounds of the present invention may be administered as prodrugs. In one embodiment, the compounds of the present invention are prodrugs of 4'-ethynyl-2-fluoro-2'-deoxyadenosine (EFdA), described, for example, in U.S. Pat. No. 7,339,053, which is a nucleoside reverse transcriptase inhibitor having the following formula:

[0121] [ka]

[0122] Prodrugs are useful in that they are believed to be able to tailor physicochemical properties, facilitate multiple administration paradigms, and improve the pharmacokinetic and / or pharmacodynamic profile of the active parent (EFdA). For example, compared to EFdA, the prodrug may facilitate a longer-acting parenteral administration mode and / or improved antiviral durability.

[0123] Administration of the chemical entities and chemical combinations described herein can be via any of the modes of administration accepted for drugs with similar utilities, including, but not limited to, oral, sublingual, subcutaneous, intravenous, intranasal, topical, transdermal, intraperitoneal, intramuscular, pulmonary, vaginal, rectal, or intraocular. In some embodiments, oral or parenteral administration is used. Examples of administration include, but are not limited to, oral once every seven days, intramuscular once every eight weeks, or subcutaneous once every six months.

[0124] Pharmaceutical compositions or formulations include solid, semi-solid, liquid, and aerosol dosage forms, such as tablets, capsules, powders, solutions, suspensions, suppositories, aerosols, etc. Chemical entities may also be administered in sustained- or controlled-release dosage forms, such as depot injections, osmotic pumps, pills, transdermal (e.g., electrotransport) patches, etc., for prolonged administration at a predetermined rate and / or for timed pulse administration. In certain embodiments, the compositions are provided in unit dosage forms suitable for single administration of precise doses.

[0125] The chemical entities described herein may be administered alone or, more typically, in combination with conventional pharmaceutical carriers, excipients, and the like (e.g., mannitol, lactose, starch, magnesium stearate, sodium saccharin, talcum, cellulose, croscarmellose sodium, glucose, gelatin, sucrose, magnesium carbonate, and the like). If desired, the pharmaceutical compositions can also contain minor amounts of nontoxic auxiliary substances, such as wetting agents, emulsifying agents, solubilizing agents, pH buffering agents, and the like (e.g., sodium acetate, sodium citrate, cyclodextrin derivatives, sorbitan monolaurate, triethanolamine acetate, triethanolamine oleate, and the like). Generally, depending on the intended mode of administration, pharmaceutical compositions contain about 0.005 to 95% by weight of the chemical entities; in certain embodiments, about 0.5 to 50% by weight. Actual methods for preparing such dosage forms are known or will be apparent to those skilled in the art. See, e.g., Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pennsylvania.

[0126] In certain embodiments, the composition takes the form of a pill or tablet, and thus comprises the active ingredient together with diluents such as lactose, sucrose, dicalcium phosphate, etc.; lubricants such as magnesium stearate, etc.; and binders such as starch, acacia gum, polyvinylpyrrolidine, gelatin, cellulose, cellulose derivatives, etc. In another solid dosage form, a powder, marume, solution, or suspension (e.g., in propylene carbonate, vegetable oils, or triglycerides) is encapsulated in a gelatin capsule.

[0127] For example, pharmaceutically administrable liquid compositions can be prepared by dissolving, dispersing, or otherwise forming a solution or suspension of at least one chemical and optional pharmaceutical adjuvants in a carrier (e.g., water, saline, aqueous dextrose, glycerol, glycol, ethanol, etc.). Injectables can be prepared in conventional forms, such as liquid solutions or suspensions, emulsions, or solid forms suitable for dissolution or suspension in liquid prior to injection. The percentage of chemical contained in such parenteral compositions will largely depend on the specific nature of the chemical, as well as the activity of the chemical and the needs of the subject. However, the percentage of active ingredient can be used in solution at 0.01% to 10%, and higher percentages of active ingredient can be used when the composition is a solid that is subsequently diluted to the above percentages. In certain embodiments, the composition may contain about 0.2 to 2% active ingredient in solution.

[0128] Pharmaceutical compositions of the chemical entities described herein may also be administered to the respiratory tract as an aerosol or solution for a nebulizer, or as a microfine powder for insufflation, either alone or in combination with an inert carrier, such as lactose, In such cases, the particles of the pharmaceutical composition have diameters of less than 50 μm, and in certain embodiments, less than 10 μm.

[0129] Generally, the chemical substance provided is administered in a therapeutically effective amount by any of the accepted modes of administration for drugs with similar utilities. The actual amount of the chemical substance, i.e., the active ingredient, depends on many factors, such as the severity of the disease being treated, the age and relative health of the subject, the potency of the chemical substance used, the route of administration, the dosage form, and other factors. The drug can be administered once or more times a day, for example, once or twice a day.

[0130] Generally, chemicals are administered as pharmaceutical compositions by any one of the following routes: oral administration, systemic administration (e.g., transdermal, intranasal, or via suppository), or parenteral administration (e.g., intramuscular, intravenous, or subcutaneous). In certain embodiments, oral administration may be used with a convenient daily dosage regimen that can be adjusted according to the severity of the affliction. The composition may take the form of a tablet, pill, capsule, semisolid, powder, sustained-release formulation, solution, suspension, elixir, aerosol, or any other suitable composition. Another mode for administering provided chemicals is inhalation.

[0131] The choice of formulation depends on various factors, such as the mode of drug administration and the bioavailability of the drug substance. For delivery by inhalation, chemicals can be formulated as liquid solutions, suspensions, aerosol propellants, or dry powders and loaded into a suitable dispenser for administration. Several types of pharmaceutical inhalation devices exist: nebulizer inhalers, metered-dose inhalers (MDIs), and dry-powder inhalers (DPIs). Nebulizer devices produce a high-velocity air stream that causes the therapeutic agent (formulated in liquid form) to spray as a mist that is carried to the patient's respiratory tract. MDIs are typically formulations packaged with compressed gas. Upon actuation, the device releases a measured amount of therapeutic agent via compressed gas, thus providing a reliable method of administering a set amount of medication. DPIs dispense the therapeutic agent in the form of a free-flowing powder, which the device can disperse into the patient's inspiratory airstream upon breathing. To achieve a free-flowing powder, the therapeutic agent is formulated with an excipient, such as lactose. A measured amount of therapeutic agent is stored in a capsule form and dispensed with each actuation.

[0132] In recent years, pharmaceutical compositions for drugs with poor bioavailability have been developed based on the principle that bioavailability can be increased by increasing the surface area, i.e., by reducing particle size. For example, U.S. Patent No. 4,107,288 describes a pharmaceutical formulation having particles in the size range of 10 to 1,000 nm in which the active substance is supported on a crosslinked polymer matrix. U.S. Patent No. 5,145,684 describes the preparation of a pharmaceutical formulation in which the drug substance is milled into nanoparticles (average particle size 400 nm) in the presence of a surface modifier and then dispersed in a liquid medium to obtain a pharmaceutical formulation with significantly higher bioavailability.

[0133] The compositions generally comprise at least one chemical entity described herein in combination with at least one pharmaceutically acceptable excipient. Acceptable excipients are non-toxic, aid in administration, and do not adversely affect the therapeutic benefit of at least one chemical entity described herein. Such excipients may be any solid, liquid, semi-solid, or, in the case of aerosol compositions, gaseous excipients generally available to those skilled in the art.

[0134] Solid pharmaceutical excipients include starch, cellulose, talc, glucose, lactose, sucrose, gelatin, malt, rice, wheat flour, chalk, silica gel, magnesium stearate, sodium stearate, glycerol monostearate, sodium chloride, dried skim milk, etc. Liquid and semi-solid excipients may be selected from glycerol, propylene glycol, water, ethanol, and various oils, such as those of petroleum, animal, vegetable, or synthetic origin, for example, peanut oil, soybean oil, mineral oil, sesame oil, etc. Liquid carriers for injectable solutions include water, saline, aqueous dextrose, and glycols.

[0135] Compressed gases may be used to disperse the chemical entities described herein in aerosol form. Suitable inert gases for this purpose include nitrogen, carbon dioxide, and the like. Other suitable pharmaceutical excipients and their formulations are described in Remington's Pharmaceutical Sciences, edited by E. W. Martin (Mack Publishing Company, 18th ed., 1990).

[0136] The amount of chemicals in the composition can vary within the full range used by those skilled in the art. Typically, the composition will contain, on a weight percent (wt%) basis, about 0.01 to 99.99 wt% of at least one chemical described herein based on the total composition, with the remainder being one or more suitable pharmaceutical excipients. In certain embodiments, at least one chemical described herein is present at a level of about 1 to 80 wt%.

[0137] In various embodiments, the pharmaceutical compositions of the present invention include compounds of formula (I), salts thereof, and combinations of the above.

[0138] Synthesis method The synthetic methods may employ readily available starting materials using the following general methods and procedures. Where typical or preferred process conditions (i.e., reaction temperatures, times, molar ratios of reactants, solvents, pressures, etc.) are given, it is understood that other process conditions can also be used unless otherwise stated. Optimum reaction conditions may vary with the particular reactants or solvents used, but such conditions can be determined by one skilled in the art by routine optimization procedures.

[0139] Furthermore, the methods of the present invention can use protecting groups that prevent certain functional groups from undergoing undesired reactions. Suitable protecting groups for various functional groups, as well as suitable conditions for protecting and deprotecting particular functional groups, are well known in the art. For example, numerous protecting groups are described in T.W. Greene and G.M.Wuts, Protecting Groups in Organic Synthesis, 3rd Edition, Wiley, New York, 1999, and references cited therein.

[0140] Additionally, provided chemical compounds may contain one or more chiral centers, and such compounds can be prepared or isolated as pure stereoisomers, i.e., as separate enantiomers or diastereomers, or as stereoisomer-enriched mixtures. All such stereoisomers (and enriched mixtures) are encompassed within the scope of this specification, unless otherwise indicated. Pure stereoisomers (or enriched mixtures) can be prepared using, for example, optically active starting materials or stereoselective reagents well-known in the art. Alternatively, racemic mixtures of such compounds can be separated using, for example, chiral column chromatography, chiral resolving agents, etc.

[0141] The compounds of formula (I) herein, including those in Examples 1-16, contain a phosphorus chiral center. The isomeric mixtures in each of Examples 1-16 were separated to obtain isomer #A, e.g., isomer 1A (the faster-eluting isomer), and isomer #B, e.g., isomer 1B (the slower-eluting isomer), based on their elution order observed in the separations performed in the Examples. Retention times, if provided, are provided solely to indicate the relative elution order of each isomer in the Examples. The elution order of the separated isomers may differ if performed under conditions different from those used herein. The absolute stereochemistry (R or S) of the phosphorus chiral center in each of the "A" and "B" stereoisomers in Examples 1-16 was not determined. Asterisks ( *) is sometimes used to indicate the chiral center at phosphorus in the relevant chemical structure diagrams of the example compounds.

[0142] The starting materials for the following reactions are generally known compounds or can be prepared by known procedures or obvious modifications thereof. For example, many of the starting materials are available from commercial sources, such as Aldrich Chemical Co. (Milwaukee, Wisconsin, USA), Bachem (Torrance, California, USA), Ernka-Chemce, or Sigma (St. Louis, Missouri, USA). Other starting materials can be prepared by procedures described in standard reference texts, such as Fieser and Fieser's Reagents for Organic Synthesis, Volumes 1-15 (John Wiley and Sons, 1991), Rodd's Chemistry of Carbon Compounds, Volumes 1-5 and supplements (Elsevier Science Publishers, 1989), Organic Reactions, Volumes 1-40 (John Wiley and Sons, 1991), March's Advanced Organic Chemistry (John Wiley and Sons, 4th Edition), and Larock's Comprehensive Organic Transformations (VCH Publishers Inc., 1989), or obvious modifications thereof.

[0143] Unless stated to the contrary, reactions described herein may be carried out at atmospheric pressure, generally within a temperature range of −78° C. to 200° C. Furthermore, unless used in the examples or otherwise specified, reaction times and conditions are intended to be approximate, e.g., at approximately atmospheric pressure, within a temperature range of about −78° C. to about 110° C., for about 1 to about 24 hours, with the reaction being carried out overnight, averaging about 16 hours.

[0144] The terms "solvent," "organic solvent," and "inert solvent" each mean a solvent that is inert under the conditions of the reaction being described therewith, such as, for example, benzene, toluene, acetonitrile, tetrahydrofuranyl ("THF"), dimethylformamide ("DMF"), chloroform, methylene chloride (or dichloromethane or DCM), diethyl ether, methanol, N-methylpyrrolidone ("NMP"), pyridine, and the like.

[0145] Isolation and purification of the chemicals and intermediates described herein, if desired, can be carried out by any suitable separation or purification procedure, such as filtration, extraction, crystallization, column chromatography, thin layer chromatography, thick layer chromatography, or a combination of these procedures. Specific illustrations of suitable separation and isolation procedures can be found in the examples herein below. However, other equivalent separation or isolation procedures can also be used.

[0146] If desired, the (R)- and (S)-isomers can be resolved by methods known to those skilled in the art, for example, by the formation of diastereomeric salts or complexes that are separable (e.g., by crystallization); through the formation of diastereomeric derivatives that are separable (e.g., by crystallization, gas-liquid chromatography, or liquid chromatography); by the selective reaction of one enantiomer with an enantiomer-specific reagent (e.g., enzymatic oxidation or reduction) followed by separation of the altered and unaltered enantiomers; or by gas-liquid chromatography or liquid chromatography in a chiral environment (e.g., on a chiral support, e.g., silica with bound chiral ligands, or in the presence of a chiral solvent). Alternatively, specific enantiomers can be synthesized by asymmetric synthesis using optically active reagents, substrates, catalysts, or solvents, or by converting one enantiomer to the other by asymmetric transformation. [Example]

[0147] Examples and General Synthesis The following examples and prophetic synthetic methods serve to more fully describe how to make and use the above-described invention. It is understood that they in no way serve to limit the true scope of the invention, but rather are presented for illustrative purposes. Unless otherwise specified, the following abbreviations have the following meanings. If an abbreviation is not defined, it has its generally accepted meaning.

[0148] aq. = aqueous μL = microliter μM = micromolar NMR=nuclear magnetic resonance Boc = tert-butoxycarbonyl br=broad Cbz = benzyloxycarbonyl d = doublet ℃ = Celsius DCM = dichloromethane dd = double doublet DIPEA = N,N-diisopropylethylamine DMAP = N,N-dimethylaminopyridine DMF = N,N-dimethylformamide DMSO = dimethyl sulfoxide EDC = N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride EtOAc = ethyl acetate g = grams h or hr = hours HPLC = High-Performance Liquid Chromatography Hz = Hertz IU = International Units I C 50 =50% inhibitory concentration J = coupling constant (Hz) LCMS = Liquid Chromatography Mass Spectrometry m=multiplet M = molar concentration M+H = parent mass spectrum peak plus H mg = milligram min=minutes mL = milliliters mM = millimolar concentration mm = millimeters mmol = millimolar MMTr = monomethoxytrityl MS = mass spectrum MTBE = methyl tert-butyl ether nM = nanomolar PE = petroleum ether ppm=parts per million qs=sufficient amount s = singlet RT=retention time sat.=saturated t = triplet TBDMS = tert-butyldimethylsilyl TBDPS = tert-butyldiphenylsilyl TEA = triethylamine THF = tetrahydrofuran TMS = trimethylsilyl

[0149] Additionally, various compounds of the present invention can be made, in one embodiment, by the general synthetic routes set forth in Schemes 1-2 below.

[0150] Scheme 1

[0151] [ka]

[0152] Scheme 2

[0153] [ka]

[0154] Scheme 3

[0155] [ka]

[0156] Scheme 4

[0157] [ka]

[0158] The absolute stereochemistry (R or S) of the phosphorus chiral center in each of the "A" and "B" stereoisomers of Schemes 1, 2, 3 and 4 was not determined.

[0159] In Schemes 1, 2, 3, and 4, n is 1 to 10, and R 1 , R 2 , R 3 and R 4 each of which is alkyl, alkylene, aryl, or aryl; 1 , R 2 , R 3 and R 4 Each of the groups independently represents one or more (C1-C 14 ) alkyl, Cl, F, oxo or (C1-C6) alkoxy.

[0160] Example 1

[0161] [ka]

[0162] Step 1: Hexadecyl (tert-butoxycarbonyl)-L-phenylalaninate To a mixture of hexadecan-1-ol (15 g, 61.9 mmol), (tert-butoxycarbonyl)-L-phenylalanine (16.41 g, 61.9 mmol), 1H-imidazole (12.64 g, 186 mmol), and HATU (35.3 g, 93 mmol) in DCM (200 mL) was added DIPEA (32.4 mL, 186 mmol). The mixture was stirred at room temperature for 16 h. TLC (petroleum ether: EtOAc = 20:1, Rf = 0.5) indicated the reaction was complete. The mixture was washed with water, dried over Na2SO4, and concentrated to give the crude product. The crude product was purified using flash silica gel chromatography (petroleum ether: EtOAc = 20:1) to give hexadecyl (tert-butoxycarbonyl)-L-phenylalaninate (23 g, 47.0 mmol, 76% yield) as a white solid.

[0163] Step 2: Hexadecyl L-phenylalaninate To a solution of hexadecyl (tert-butoxycarbonyl)-L-phenylalaninate (23 g, 47.0 mmol) in DCM (500 mL) was added trifluoroacetic acid (100 mL). The resulting mixture was stirred at room temperature for 12 h. TLC (petroleum ether: EtOAc = 10:1, RF = 0.3) indicated the reaction was complete. The reaction mixture was concentrated to remove DCM, and the pH of the residue was adjusted to approximately 8 with NaHCO3 (aq) and extracted twice with EtOAc (100 mL). The organic phase was washed with water (100 mL), brine (100 mL), dried over Na2SO4, and concentrated to give the crude product. The crude product was purified using flash silica gel chromatography (petroleum ether: EtOAc = 5:1) to give hexadecyl L-phenylalaninate (18 g, 46.2 mmol, 98% yield) as a white solid. 1H NMR (400 MHz, DMSO) δ 7.26 (t, J = 7.4 Hz, 2H), 7.18 (t, J = 8.4 Hz, 3H), 4.09 - 3.85 (m, 2H), 3.66 (t, J = 6.8 Hz, 1H), 2.89 - 2.80 (m, 2H), 1.45 (dd, J = 13.2, 6.5 Hz, 2H), 1.22 (s, 26H), 0.84 (t, J = 6.7 Hz, 3H).

[0164] Step 3: Hexadecyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a cold (ice-water bath) solution of hexadecyl L-phenylalaninate (2.66 g, 6.82 mmol) and triethylamine (1.188 mL, 8.52 mmol) in DCM (6 mL), a solution of phenyl phosphorodichloridate (1.799 g, 8.52 mmol) in DCM (1 mL) was added dropwise under a nitrogen atmosphere. The reaction mixture was then stirred at room temperature for 1 hour. To a cold (ice-water bath) solution of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (1 g, 3.41 mmol) in THF (20 mL) and pyridine (10 mL), tert-butylmagnesium chloride (8.52 mL, 8.52 mmol) was added dropwise under a nitrogen atmosphere. The mixture was then stirred at room temperature for 30 minutes. The first prepared mixture was added dropwise to the second prepared mixture at room temperature and stirred for 2 hours. LCMS showed the presence of the product. The mixture was filtered and concentrated to give a yellow solid. The solid was then dissolved in EtOAc (40 mL) and partitioned with HCl (0.05 N, 20 mL). The layers were separated and the aqueous layer was extracted with EtOAc (40 mL). The combined organic phase was washed with brine, dried over Na2SO4, filtered, and concentrated to give the crude product. The residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 0-100% CH3CN / 10 mM NH4HCO3 in water) to give hexadecyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (1.3 g, 0.091 mmol, 2.66% yield) as a yellow solid. LCMS (M+H)=821.0; Retention time (0.1%TFA)=3.147 minutes.

[0165] Step 4: (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((((S)-1-(hexadecyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl decanoate To a solution of decanoic acid (185 mg, 1.072 mmol) in DCM (60 mL) were added DMAP (595 mg, 4.87 mmol) and EDC (934 mg, 4.87 mmol), and the resulting mixture was stirred at room temperature for 0.5 h. Then, hexadecyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (800 mg, 0.974 mmol) was added, and the resulting mixture was stirred at room temperature overnight. LCMS showed the reaction was complete. The reaction mixture was quenched with water (40 mL) and extracted with DCM (30 mL × 3). The combined organic phase was washed with brine (30 mL), dried over Na2SO4, and concentrated in vacuo. The residue was purified using a reverse-phase column (Boston, 120 g, 0-100% CHCN / 10 mM NHHCO in water) to give (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((((S)-1-(hexadecyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl decanoate (400 mg, 0.407 mmol, 41.7% yield). LCMS: Retention time (0.1% TFA) = 4.626 min. The diastereomers were separated by preparative SFC (instrument: SFC-80 (Thar, Waters); column: OZ 20 × 250 mm, 10 μm (Daicel); column temperature: 40 °C; mobile phase: CO / MEOH (0.5% ammonia / methanol) = 45 / 55; flow rate: 80 g / min; back pressure: 100 bar; detection wavelength: 260 nm; cycle time: 10 min; sample solution: 430 mg dissolved in 30 mL methanol; injection volume: 2.2 mL) to give the first-eluting isomer (Example 1A, RT: 1.62 min, 130 mg, 0.126 mmol, 30.7% yield) as a white solid, followed by the second-eluting isomer (Example 1B, RT: 2.68 min, 135 mg, 0.136 mmol, 33.2% yield) as a white solid. Isomer eluting first: LCMS: retention time (0.1% TFA) = 4.714 min. HPLC: retention time (0.1% NH4HCO3) = 15.833 min. 1 H NMR (400 MHz, CDCl3) δ 8.04 (s, 1H), 7.28 (d, J = 7.7 Hz, 1H), 7.25 - 7.19 (m, 4H), 7.16 (d, J = 8.6 Hz, 2H), 7.12 (t, J = 7.6 Hz, 1H), 7.03 (dd, J = 7.5, 1.8 Hz, 2H), 6.43 (t, J = 6.9 Hz, 1H), 6.09 (brs, 2H), 5.58 (dd, J = 6.7, 3.8 Hz, 1H), 4.29 (dd, J = 11.1, 6.7 Hz, 1H), 4.23 (ddd, J = 15.1, 9.8, 5.3 Hz, 1H), 4.11 (dd, J = 11.1, 5.9 Hz, 1H), 4.01 (td, J = 6.8, 2.0 Hz, 2H), 3.71 - 3.60 (m, 1H), 2.97 (d, J = 6.3 Hz, 2H), 2.64 - 2.48 (m, 3H), 2.38 (t, J = 7.6 Hz, 2H), 1.82 (s, 2H), 1.70 - 1.60 (m, 2H), 1.25 (t, J = 7.7 Hz, 38H), 0.87 (dt, J = 4.1, 3.3 Hz, 6H) Isomer eluting second: LCMS: retention time (0.1% TFA) = 4.774 min. HPLC: retention time (0.1% NH4HCO3) = 14.483 min. 1H NMR (400 MHz, CDCl3) δ 8.00 (s, 1H), 7.21 (ddd, J = 10.7, 6.9, 5.3 Hz, 5H), 7.15 (d, J = 8.6 Hz, 2H), 7.13-7.08 (m, 3H), 6.39 (t, J = 6.7 Hz, 1H), 6.12 (brs, 2H), 5.56 (dd, J = 6.5, 4.4 Hz, 1H), 4.36-4.28 (m, 1H), 4.25 (dd, J = 11.1, 6.3 Hz, 1H), 4.13 (dd, J = 11.1, 5.7 Hz, 1H), 4.02 (t, J = 6.8 Hz, 2H), 3.69 (t, J = 10.7 Hz, 1H), 3.00 (d, J = 6.5 Hz, 2H), 2.68-2.53 (m, 3H), 2.40 (t, J = 7.6 Hz, 2H), 1.82 (s, 4H), 1.71-1.62 (m, 2H), 1.26 (t, J = 6.8 Hz, 36H), 0.87 (dt, J = 7.0, 3.3 Hz, 6H)

[0166] Example 2

[0167]

change

[0168] Project 1: Turret To a solution of octadecan-1-ol (32.5 g, 120 mmol), (tert-butoxycarbonyl)-L-phenylalanine (29 g, 109 mmol), and HATU (62.3 g, 164 mmol) in DCM (100 mL) was added 1H-imidazole (22.32 g, 328 mmol) and DIPEA (57.3 mL, 328 mmol). The resulting mixture was stirred at 25 °C overnight. TLC (petroleum ether: EtOAc = 10:1, Rf = 0.5) showed the reaction was complete. The reaction was quenched with water (200 mL) and DCM (100 mL). The organic layer was then separated, and the aqueous layer was extracted with DCM (2 × 50 mL). The combined organic layers were washed with brine (200 mL), dried over Na SO , filtered, and concentrated to give the crude product. The residue was purified using combiflash (330 g silica gel column, hexane:ethyl acetate=20:1). Appropriate fractions containing the desired product were combined and concentrated in vacuo to give octadecyl (tert-butoxycarbonyl)phenylalaninate (32 g, 61.8 mmol, 56.5% yield) as a yellow solid.

[0169] Step 2: Octadecyl phenylalaninate To a solution of octadecyl (tert-butoxycarbonyl)phenylalaninate (32 g, 61.8 mmol) in DCM (400 mL) was added TFA (28.6 mL, 371 mmol). The resulting mixture was stirred at 25 °C for 2 h. TLC (petroleum ether: EtOAc = 10:1, Rf = 0.1) showed the reaction was complete. The reaction was quenched with 1 N NaHCO (200 mL) and DCM (100 mL), and then the organic layer was separated. The aqueous layer was extracted with DCM (2 × 100 mL). The combined organic layers were washed with brine (200 mL), dried over NaSO, and concentrated to give crude octadecyl phenylalaninate (19 g, 45.5 mmol, 73.6% yield), which was used in the next step without purification.

[0170] Step 3: Octadecyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)phenylalaninate To a solution of octadecyl L-phenylalaninate (5.70 g, 13.64 mmol) and triethylamine (1.901 mL, 13.64 mmol) in DCM (80 mL) was added dropwise a solution of phenyl phosphorodichloridate (2.038 mL, 13.64 mmol) in DCM (80 mL) at approximately 5°C under a nitrogen atmosphere. The reaction mixture was then stirred at room temperature for 1 hour. To a solution of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (2 g, 6.82 mmol) in THF (40 mL) and pyridine (20 mL) was added tert-butylmagnesium chloride (17.05 mL, 17.05 mmol) dropwise under a nitrogen atmosphere at 0°C. The mixture was then stirred at room temperature for 30 minutes. The first prepared mixture was added dropwise to the second prepared mixture at room temperature and stirred for 2 hours. LCMS showed the reaction was complete. The mixture was filtered and concentrated to give a yellow solid. The solid was then dissolved in EtOAc (40 mL) and partitioned with HCl (0.05 N, 20 mL). The layers were separated and the aqueous layer was extracted with EtOAc (40 mL). The combined organic phase was washed with brine, dried over Na2SO4, filtered, and concentrated to give the crude product. The residue was purified using reverse-phase chromatography (SepaFlash® C18, BOSTON, 0–100% CH3CN / 10 mM NH4HCO3 in water) to give octadecyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)phenylalaninate (1.2 g, 1.354 mmol, 19.85% yield) as a yellow solid. LCMS (M+H) = 849.4; retention time (0.1% TFA) = 3.568 min.The diastereomers were separated by preparative SFC (instrument: SFC-150 (Waters); column: IH 20 × 250 mm, 10 μm (Daicel); column temperature: 35 °C; mobile phase: CO2 / MeOH (0.2% ammonia / methanol) = 60 / 40; flow rate: 100 g / min; back pressure: 100 bar; detection wavelength: 214 nm; cycle time: 4 min; sample solution: 1,900 mg dissolved in 100 mL methanol; injection volume: 3 mL) to give the first-eluting isomer (Intermediate 2A, RT: 1.76 min, 800 mg, 0.908 mmol, 38.5% yield) and the second-eluting isomer (Intermediate 2B, RT: 3.21 min, 650 mg, 0.766 mmol, 32.5% yield) as white solids. First eluting isomer: LCMS (M+H) = 849.4; Retention time (0.1% TFA) = 2.874 min. HPLC: Retention time (0.1% NH4HCO3): 11.004 min. 1 H NMR (400MHz, CDCl3) δ 7.92 (s, 1H), 7.25-7.17 (m, 5H), 7.14 (d, J = 7.3 Hz, 1H), 7.12-7.08 (m, 4H), 6.31 (dd, J = 7.3, 4.3 Hz, 1H), 6.10 (s, 2H), 4.71 (t, J = 7.6 Hz, 1H), 4.26 (dd, J = 11.2, 7.7 Hz, 2H), 4.20 (dd, J = 11.3, 8.8 Hz, 1H), 4.03 (t, J = 6.7 Hz, 2H), 3.88 (s, 1H), 2.99 (d, J = 6.2 Hz, 2H), 2.76-2.63 (m, 3H), 1.86 (s, 2H), 1.24 (d, J = 9.0 Hz, 30H), 0.88 (t, J = 6.8 Hz, 3H). Second eluting isomer: LCMS (M+H) = 849.5; Retention time (0.1% TFA) = 2.839 min. HPLC: Retention time (0.1% NH4HCO3): 10.931 min. 1H NMR (400MHz, CDCl3) δ 7.78 (s, 1H), 7.30-7.26 (m, 2H), 7.23 (dd, J = 8.4, 4.6 Hz, 3H), 7.15 (d, J = 7.8 Hz, 3H), 7.06 (dd, J = 7.5, 1.7 Hz, 2H), 6.32 (dd, J = 7.4, 4.2 Hz, 1H), 6.24 (s, 2H), 4.64 (t, J = 7.6 Hz, 1H), 4.23-4.15 (m, 2H), 4.02 (t, J = 6.7 Hz, 2H), 3.97 (dd, J = 11.5, 8.4 Hz, 1H), 3.73 (t, J = 10.8 Hz, 1H), 3.04-2.92 (m, 2H), 2.70 (s, 1H), 2.66 (dt, J = 13.7, 7.8 Hz, 1H), 2.57 (ddd, J = 13.6, 7.1, 4.2 Hz, 1H), 1.89 (s, 2H), 1.24 (d, J = 4.9 Hz, 30H), 0.87 (t, J = 6.8 Hz, 3H).

[0171] Step 4: (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((((S)-1-(octadecyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl decanoate To a solution of decanoic acid (67.0 mg, 0.389 mmol) in DCM (10 mL) were added DMAP (216 mg, 1.767 mmol) and EDC (339 mg, 1.767 mmol), and the resulting mixture was stirred at room temperature for 0.5 h. Then, octadecyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (Intermediate 2A, 300 mg, 0.353 mmol) was added, and the resulting mixture was stirred at room temperature overnight. TLC showed that the reaction was complete. The reaction mixture was diluted with DCM (60 mL), washed with water, washed with brine, dried over Na2SO4, filtered, and concentrated. The residue was purified using preparative thin-layer chromatography (DCM / MeOH=10 / 1) to give (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((((S)-1-(octadecyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl decanoate (Example 2A, 109 mg, 0.104 mmol, 29.5% yield) as a colorless oil. HPLC retention time (10 mM NH4HCO3)=15.68 min. 1H NMR (400MHz, CDCl3) δ 8.01 (s, 1H), 7.25-7.19 (m, 4H), 7.17-7.14 (m, 3H), 7.13-7.08 (m, 3H), 6.40 (t, J = 6.4 Hz, 1H), 6.08 (brs, 2H), 5.56 (dd, J = 6.8, 4.8 Hz, 1H), 4.37-4.28 (m, 1H), 4.25 (dd, J = 11.2, 6.4 Hz, 1H), 4.13 (dd, J = 11.2, 6.0 Hz, 1H), 4.02 (t, J = 6.8 Hz, 2H), 3.68 (t, J = 10.4 Hz, 1H), 3.00 (d, J = 6.4 Hz, 2H), 2.63-2.52 (m, 3H), 2.40 (t, J = 7.2 Hz, 2H), 1.67(quint, J = 7.2 Hz, 2H), 1.52(quint, J = 6.4 Hz, 2H), 1.32-1.20 (m, 42H), 0.91-0.84 (m, 6H).

[0172] To a solution of decanoic acid (55.8 mg, 0.324 mmol) in DCM (15 mL) were added DMAP (180 mg, 1.472 mmol) and EDC (282 mg, 1.472 mmol). The resulting mixture was stirred at room temperature for 0.5 h. Then, octadecyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (Intermediate 2B, 250 mg, 0.294 mmol) was added, and the resulting mixture was stirred at room temperature overnight. TLC (DCM:MeOH = 10:1, Rf = 0.8) showed the reaction was complete. The reaction mixture was quenched with water (10 mL) and extracted with DCM (20 mL × 3). The combined organic phase was washed with brine (30 mL), dried over NaSO, and concentrated in vacuo. The mixture was purified using preparative thin-layer chromatography (DCM:CHOH=10:1, Rf=0.7) to give (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((((S)-1-(octadecyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl decanoate (Example 2B, 140 mg, 0.140 mmol, 47.4% yield). HPLC: Retention time (10 mM NH4HCO3)=16.183 min. 1H NMR (400MHz, CDCl3) δ 8.06 (s, 1H), 7.27 (d, J = 7.5 Hz, 1H), 7.25-7.20 (m, 2H), 7.17 (dd, J = 12.9, 7.0 Hz, 4H), 7.11 (d, J = 7.1 Hz, 1H), 7.03 (dd, J = 7.4, 1.8 Hz, 2H), 6.43 (t, J = 6.8 Hz, 1H), 6.14 (s, 2H), 5.58 (dd, J = 6.7, 3.8 Hz, 1H), 4.29 (dd, J = 11.0, 6.8 Hz, 1H), 4.26-4.18 (m, 1H), 4.11 (dd, J = 11.1, 5.9 Hz, 1H), 4.01 (td, J = 6.8, 1.8 Hz, 2H), 3.67 (t, J = 10.8 Hz, 1H), 2.97 (d, J = 6.4 Hz, 2H), 2.64-2.56 (m, 2H), 2.52 (ddd, J = 13.9, 6.2, 3.8 Hz, 1H), 2.38 (t, J = 7.6 Hz, 2H), 1.92 (s, 2H), 1.69-1.61 (m, 2H), 1.30-1.21 (m, 42H), 0.87 (dt, J = 7.0, 3.3 Hz, 6H).

[0173] Example 3

[0174]

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[0175] Project 1: Tert-ブトキシカルボニル-L-フェニルアラニネート A mixture of icosan-1-ol (33.8 g, 113 mmol), (tert-butoxycarbonyl)-L-phenylalanine (25 g, 94 mmol), imidazole (19.25 g, 283 mmol), DIPEA (49.4 mL, 283 mmol), and HATU (53.7 g, 141 mmol) in DCM (200 mL) was stirred at 25 °C for 16 h. TLC showed the presence of a new product. Water (200 mL) was added, and the resulting mixture was extracted with DCM (150 mL × 3). The combined organic phases were washed with brine, dried over Na2SO4, filtered, and concentrated. The residue was purified using silica gel chromatography (petroleum ether: EtOAc = 30:1) to give icosyl (tert-butoxycarbonyl)-L-phenylalaninate (20 g, 10.99 mmol, 11.67% yield) as a white solid.

[0176] Step 2: Eicosyl L-phenylalaninate To a solution of icosyl (tert-butoxycarbonyl)-L-phenylalaninate (30 g, 55.0 mmol) in DCM (200 mL) was added TFA (42.3 mL, 550 mmol) at 0 °C. After 5 min, the cold bath was removed, and the mixture was stirred at room temperature for 2 h. TLC showed the presence of a new compound. The reaction mixture was washed with water and saturated NaHCO 3 . The combined aqueous layers were extracted with DCM (200 mL × 3). The combined organic layers were washed with brine, dried over Na 2 SO 4 , filtered, and concentrated to give a residue. The residue was purified using silica gel chromatography (DCM:MeOH = 20:1) to give icosyl L-phenylalaninate (25 g, 50.5 mmol, 92% yield). 1 H NMR (400MHz, DMSO) δ 7.40-7.10 (m, 5H), 4.0-3.85 (m, 2H), 3.55 (t, J = 6.8 Hz, 1H), 2.92- 2.71 (m, 2H), 1.91 (s, 1H), 1.54-1.38 (m, 2H), 1.23 (s, 33H), 0.85 (t, J = 6.8 Hz, 3H).

[0177] Step 3: Icosyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a cooled solution of icosyl L-phenylalaninate (7.60 g, 17.05 mmol) and triethylamine (2.376 mL, 17.05 mmol) in DCM (48 mL) was added a solution of phenyl phosphorodichloridate (3.60 g, 17.05 mmol) in DCM (1 mL) dropwise under a nitrogen atmosphere at 0° C., and the reaction mixture was stirred at room temperature for 1 hour. To a cooled solution of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (2 g, 6.82 mmol) in THF (24 mL) and pyridine (12 mL) was added tert-butylmagnesium chloride (17.05 mL, 17.05 mmol) dropwise under a nitrogen atmosphere at 0° C., and the reaction mixture was stirred at room temperature for 30 minutes. The first mixture was added dropwise to the second mixture at room temperature and stirred for 2 hours. LCMS showed that the starting material was completely consumed. The reaction mixture was filtered, and the filtrate was concentrated. The residue was diluted with DCM, washed three times with 0.5 N HCl, washed with brine, dried over Na2SO4, filtered, and concentrated. The residue was purified using silica gel chromatography (DCM:MeOH = 0-5%) to give icosyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (800 mg, 0.867 mmol, 12.71% yield). LCMS (M+H) = 876.0; retention time (0.1% TFA) = 4.30 min.

[0178] Step 4: (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((((S)-1-(icosyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl decanoate To a solution of icosyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (1000 mg, 1.140 mmol) in DCM (10 mL) was added DMAP (139 mg, 1.140 mmol), DIEA (0.597 mL, 3.42 mmol), and EDC (1093 mg, 5.70 mmol). The resulting mixture was stirred at room temperature for 0.5 h. Decanoic acid (196 mg, 1.140 mmol) was then added, and the reaction was stirred at room temperature overnight. TLC indicated the presence of a new product. The reaction mixture was diluted with water and extracted with DCM (100 mL × 3). The combined organic phase was washed with brine, dried over Na2SO4, filtered, and concentrated to give a residue. The residue was purified using silica gel chromatography (DCM:MeOH=10:1) to give (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((((S)-1-(icosyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl decanoate (500 mg, 0.446 mmol, 39.1% yield). The diastereomers were separated by preparative SFC (SFC-80 (Thar, Waters), column: (R,R) Whelk-O1 20 × 250 mm, 10 μm (Daicel), column temperature: 40 °C, mobile phase: CO / MEOH (0.2% ammonia / methanol) = 45 / 55, flow rate: 80 g / min, back pressure: 100 bar, detection wavelength: 260 nm, cycle time: 9 min) to give the first-eluting isomer (Example 3A, 120 mg, 0.114 mmol, 20.93% yield) as a white solid, followed by the second-eluting isomer (Example 3B, 110 mg, 0.107 mmol, 19.64% yield) as a white solid. First eluting isomer: 1H NMR (400MHz, CDCl3) δ 8.00 (s, 1H), 7.24-6.99 (m, 10H), 6.43 (t, J = 6.9 Hz, 1H), 6.03 (s, 1H), 5.58 (dd, J = 6.7, 3.7 Hz, 1H), 4.35- 4.18 (m, 2H), 4.11 (dd, J = 11.0, 5.8 Hz, 1H), 4.02 (dd, J = 6.7, 5.1 Hz, 2H), 3.62 (t, J = 10.8 Hz, 1H), 2.96 (d, J = 6.2 Hz, 2H), 2.66- 2.49 (m, 3H), 2.38 (t, J = 7.6 Hz, 2H), 1.54 (d, J = 6.0 Hz, 3H), 1.25 (t, J = 7.1 Hz, 48H), 0.88 (dd, J = 6.8, 5.4 Hz, 6H). Chapter 2: Dissolving the heterosexual body: 1 H NMR (400MHz, CDCl3) δ 7.98 (s, 1H), 7.25-7.08 (m, 10H), 6.40 (t, J = 6.7 Hz, 1H), 6.02 (s, 1H), 5.56 (dd, J = 6.5, 4.5 Hz, 1H), 4.38-4.07 (m, 3H), 4.05-3.97 (m, 2H), 3.64 (t, J = 10.7 Hz, 1H), 3.09-2.93 (m, 2H), 2.67-2.55 (m, 3H), 2.40 (t, J = 7.6 Hz, 2H), 1.58-1.45 (m, 3H), 1.27 (d, J = 9.3 Hz, 48H), 0.93- 0.84 (m, 6H).

[0179] Example 4

[0180]

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[0181] Project 1: Tert-ブトキシカルボニル-L-フェニルアラニネート A mixture of docosan-1-ol (55.4 g, 170 mmol), (tert-butoxycarbonyl)-L-phenylalanine (50 g, 188 mmol), imidazole (38.5 g, 565 mmol), DIPEA (99 mL, 565 mmol), and HATU (107 g, 283 mmol) in DCM (500 mL) was stirred at 25 °C for 16 h. TLC showed the presence of a new compound. Water (500 mL) was added, and the mixture was extracted with DCM (250 mL × 3). The combined organic phases were washed with brine, dried over Na2SO4, and concentrated. The residue was purified using silica gel chromatography (petroleum ether: EtOAc = 20:1) to give docosyl (tert-butoxycarbonyl)-L-phenylalaninate (48 g, 79 mmol, 42.2% yield) as a white solid. 1 H NMR (400MHz, CDCl3) δ 7.33-7.23 (m, 3H), 7.13 (d, J = 7.0 Hz, 2H), 4.98 (d, J = 7.9 Hz, 1H), 4.57 (d, J = 7.0 Hz, 1H), 4.18-4.01 (m, 2H), 3.20- 2.96 (m, 2H), 1.68- 1.52 (m, 2H), 1.42 (s, 9H), 1.26 (s, 38H), 0.88 (t, J = 6.8 Hz, 3H).

[0182] Step 2: Docosyl L-phenylalaninate To a cold (ice-water bath) solution of docosyl (tert-butoxycarbonyl)-L-phenylalaninate (48 g, 84 mmol) in DCM (500 mL) was added TFA (64.4 mL, 836 mmol). After 5 min, the mixture was removed from the cold bath and stirred at room temperature for 2 h. TLC indicated the reaction was complete. The reaction mixture was washed with water and saturated Na2CO3. The combined aqueous layer was extracted with DCM (400 mL × 3), and the combined organic layer was washed with brine, dried over Na2SO4, filtered, and concentrated to give a residue. The residue was purified using silica gel chromatography (DCM:MeOH = 20:1) to give docosyl L-phenylalaninate (36 g, 72.2 mmol, 86% yield). 1H NMR (400MHz, DMSO) δ 7.40-7.10 (m, 5H), 4.07- 3.85 (m, 2H), 3.55 (t, J = 6.8 Hz, 1H), 2.92- 2.71 (m, 2H), 1.91 (s, 1H), 1.54-1.38 (m, 2H), 1.23 (s, 38H), 0.85 (t, J = 6.8 Hz, 3H).

[0183] Step 3: Docosyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a cold (ice-water bath) solution of docosyl L-phenylalaninate (2019 mg, 4.26 mmol) and triethylamine (0.238 mL, 1.705 mmol) in DCM (48 mL), a solution of phenyl phosphorodichloridate (899 mg, 4.26 mmol) in DCM (2 mL) was added dropwise under a nitrogen atmosphere, and the reaction mixture was stirred at room temperature for 1 h. To a cold (ice-water bath) solution of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (500 mg, 1.705 mmol) in THF (20 mL) / pyridine (10 mL), tert-butylmagnesium chloride (3.41 mL, 3.41 mmol) was added dropwise under a nitrogen atmosphere, and the mixture was stirred at room temperature for 30 min. The first mixture was added dropwise to the second mixture at room temperature and stirred for 2 hours. LCMS indicated the reaction was complete. The reaction mixture was filtered, and the filtrate was concentrated. The residue was diluted with DCM, washed three times with 0.5 N HCl, washed with brine, dried over Na2SO4, filtered, and concentrated to give a residue. The residue was purified using flash chromatography (0–5% MeOH / DCM) to give docosyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (400 mg, 0.442 mmol, 25.9% yield). LCMS (M+H) = 905.7; retention time (0.1% TFA) = 4.39 min.The diastereomers were separated by preparative SFC (SFC-80 (Thar, Waters), column: (R,R) Whelk-O1 20 × 250 mm, 10 μm (Daicel), column temperature: 40 °C, mobile phase: CO2 / MEOH (0.2% ammonia / methanol) = 45 / 55, flow rate: 80 g / min, back pressure: 100 bar, detection wavelength: 260 nm, cycle time: 9 min), and the first-eluting isomer, docosyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (Intermediate 4A, 1.2 g, 1.326 mmol, 40.0% yield) was obtained as a white solid (LCMS (M+H)=906.7), followed by the second-eluting isomer docosyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (Intermediate 4B, 1.5 g, 1.657 mmol, 50.0% yield) (LCMS (M+H)=906.7).

[0184] Step 4: (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((((S)-1-(docosyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl decanoate To a solution of docosyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (Intermediate 4A, 400 mg, 0.442 mmol) in DCM (10 mL) was added DMAP (54.0 mg, 0.442 mmol), EDC (254 mg, 1.326 mmol), and DIEA (0.232 mL, 1.326 mmol). The resulting mixture was stirred at room temperature for 0.5 h. Decanoic acid (76 mg, 0.442 mmol) was then added, and the resulting mixture was stirred at room temperature overnight. TLC showed the presence of a new compound. The reaction mixture was diluted with water and extracted with DCM (20 mL × 3). The combined organic phase was washed with brine, dried over Na2SO4, filtered, and concentrated to a residue that was purified using flash chromatography (0 to 10% MeOH / DCM) to give (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((((S)-1-(docosyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl decanoate (Example 4A, 260 mg, 0.239 mmol, 54.0% yield). 1H NMR (400MHz, CDCl3) δ 7.98 (s, 1H), 7.26 - 7.05 (m, 10H), 6.40 (t, J = 6.7 Hz, 1H), 6.09 (s, 1H), 5.56 (dd, J = 6.5, 4.4 Hz, 1H), 4.39-4.19 (m, 2H), 4.13 (dd, J = 11.1, 5.6 Hz, 1H), 4.02 (t, J = 6.8 Hz, 2H), 3.76-3.60 (m, 1H), 3.11-2.94 (m, 2H), 2.67-2.57 (m, 3H), 2.39 (d, J = 7.7 Hz, 2H), 1.66 (dd, J = 14.7, 7.3 Hz, 2H), 1.50 (dd, J = 20.8, 14.5 Hz, 2H), 1.36-1.20 (m, 51H), 0.93- 0.78 (m, 6H).

[0185] To a solution of docosyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (Intermediate 4B, 400 mg, 0.442 mmol) in DCM (10 mL) was added DMAP (54.0 mg, 0.442 mmol), EDC (254 mg, 1.326 mmol), and DIEA (0.232 mL, 1.326 mmol). The resulting mixture was stirred at room temperature for 0.5 hours. Decanoic acid (76 mg, 442 mmol) was then added, and the resulting mixture was stirred at room temperature overnight. TLC showed the presence of a new compound. The reaction mixture was diluted with water and extracted with DCM (20 mL × 3). The combined organic phase was washed with brine, dried over Na2SO4, filtered, and concentrated to a residue that was purified using flash chromatography (0-10% MeOH / DCM) to give (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((((S)-1-(docosyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl decanoate (Example 4B, 200 mg, 0.188 mmol, 42.5% yield). 1H NMR (400MHz, CDCl3) δ 8.01 (s, 1H), 7.25- 6.96 (m, 10H), 6.43 (t, J = 6.9 Hz, 1H), 6.08 (s, 1H), 5.58 (dd, J = 6.7, 3.8 Hz, 1H), 4.32-4.18 (m, 2H), 4.09 (dt, J = 14.2, 7.1 Hz, 1H), 4.06-3.94 (m, 2H), 3.67 (dt, J = 21.6, 10.9 Hz, 1H), 2.95 (t, J = 9.9 Hz, 2H), 2.66- 2.49 (m, 3H), 2.38 (t, J = 7.6 Hz, 2H), 1.64 (dd, J = 14.6, 7.3 Hz, 2H), 1.54 (d, J = 6.5 Hz, 2H), 1.38-1.17 (m, 51H), 0.97-0.81 (m, 6H).

[0186] Example 5

[0187] [ka]

[0188] (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((((S)-1-(docosyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl icosanoate To a solution of docosyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (Intermediate 4A, 400 mg, 0.442 mmol) in DCM (10 mL) was added DMAP (54.0 mg, 0.442 mmol), EDC (254 mg, 1.326 mmol), DIEA (0.232 mL, 1.326 mmol), and icosanoic acid (138 mg, 0.442 mmol). The resulting mixture was stirred at room temperature overnight. TLC indicated the presence of a new compound. The reaction mixture was diluted with water and extracted with DCM (20 mL × 3). The combined organic phase was washed with brine, dried over Na2SO4, filtered, and concentrated to give a residue. The residue was purified using flash chromatography (0-10% MeOH / DCM) to give (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((((S)-1-(docosyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl icosanoate (Example 5A, 220 mg, 0.183 mmol, 41.5% yield). 1 H NMR (400MHz, CDCl3) δ 8.02 (s, 1H), 7.25-7.03 (m, 10H), 6.40 (t, J = 6.7 Hz, 1H), 6.21 (s, 1H), 5.57 (dd, J = 6.4, 4.4 Hz, 1H), 4.42-4.21 (m, 2H), 4.14 (dd, J = 11.0, 5.7 Hz, 1H), 4.02 (t, J = 6.8 Hz, 2H), 3.75 (dt, J = 30.9, 10.5 Hz, 1H), 3.07- 2.92 (m, 2H), 2.69-2.56 (m, 3H), 2.40 (t, J = 7.6 Hz, 2H), 1.74-1.59 (m, 2H), 1.56-1.45 (m, 2H), 1.38-1.17 (m, 71H), 0.88 (t, J = 6.8 Hz, 6H).

[0189] To a solution of docosyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (Intermediate 4B, 400 mg, 0.442 mmol) in DCM (20 mL) was added DMAP (54.0 mg, 0.442 mmol), EDC (254 mg, 1,326 mmol), DIEA (0.232 mL, 1.326 mmol), and icosanoic acid (138 mg, 0.442 mmol). The resulting mixture was stirred at room temperature overnight. TLC indicated the presence of a new compound. The reaction mixture was diluted with water and extracted with DCM (20 mL × 3). The combined organic phase was washed with brine, dried over Na2SO4, filtered, and concentrated to give a residue. The residue was purified using flash chromatography (0-10% MeOH / DCM) to give (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((((S)-1-(docosyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl icosanoate (Example 5B, 160 mg, 0.133 mmol, 30.2% yield). 1H NMR (400MHz, CDCl3) δ 8.01 (s, 1H), 7.24-6.98 (m, 10H), 6.43 (t, J = 6.9 Hz, 1H), 6.10 (s, 1H), 5.59 (dd, J = 6.7, 3.7 Hz, 1H), 4.26 (ddd, J = 25.7, 13.2, 6.4 Hz, 2H), 4.11 (dd, J = 11.0, 5.8 Hz, 1H), 4.01 (dd, J = 6.7, 5.1 Hz, 2H), 3.68 (t, J = 10.8 Hz, 1H), 2.95 (t, J = 10.4 Hz, 2H), 2.66-2.49 (m, 3H), 2.38 (t, J = 7.5 Hz, 2H), 1.64 (dd, J = 14.5, 7.2 Hz, 2H), 1.50 (d, J = 17.8 Hz, 2H), 1.37-1.16 (m, 71H), 0.88 (t, J = 6.8 Hz, 6H).

[0190] Example 6

[0191]

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[0192] Project 1: tert-ブトキシカルボニル-L-フェニルアラニネート To a mixture of propan-2-ol (4.53 g, 75 mmol), (tert-butoxycarbonyl)-L-phenylalanine (20 g, 75 mmol), and HATU (34.4 g, 90 mmol) in DCM (300 mL) was added 1H-imidazole (15.40 g, 226 mmol) and triethylamine (31.4 mL, 226 mmol). The resulting mixture was stirred at 25 °C overnight. TLC showed the reaction was complete. The reaction was quenched with water (100 mL) and diluted with DCM (100 mL), and then the organic layer was separated. The aqueous layer was extracted with DCM (2 × 200 mL). The combined organic layers were washed with brine (100 mL), dried over Na SO , filtered, and concentrated. The residue was purified by silica gel column chromatography (hexane / ethyl acetate=10 / 1) to give isopropyl (tert-butoxycarbonyl)-L-phenylalaninate (22.7 g, 73.8 mmol, 98% yield) as a white solid. LCMS (M+Na) = 330.0; retention time (0.1% TFA) = 2.08 min.

[0193] Step 2: Isopropyl L-phenylalaninate To a solution of isopropyl (tert-butoxycarbonyl)-L-phenylalaninate (22.7 g, 73.8 mmol) in DCM (120 mL) was added TFA (40 mL, 519 mmol) at 0 °C. The resulting mixture was stirred at 25 °C for 4 hours. LCMS indicated the reaction was complete. The reaction mixture was concentrated under reduced pressure. The residue was neutralized with NH4HCO3 (20 mL), diluted with water (200 mL), and extracted with DCM (3 × 200 mL). The combined organic layers were then washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified using silica gel chromatography (DCM / MeOH = 10 / 1) to afford isopropyl L-phenylalaninate (12.5 g, 60.3 mmol, 82% yield) as a yellow oil. LCMS (M+H)=208.0; Retention time (0.1%TFA)=1.13 minutes.

[0194] Step 3: Isopropyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate A solution of phenyl phosphorodichloridate (3.82 mL, 25.6 mmol) in DCM (1 mL) was added dropwise to a mixture of isopropyl L-phenylalaninate (5.30 g, 25.6 mmol) and triethylamine (3.56 mL, 25.6 mmol) in DCM (60 mL) under a nitrogen atmosphere at 0°C, and the reaction mixture was stirred at the same temperature for 1 hour. To a solution of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (3 g, 10.23 mmol) in THF (20 mL) and pyridine (10 mL) was added tert-butylmagnesium chloride (25.6 mL, 25.6 mmol) dropwise under a nitrogen atmosphere at 0°C. The mixture was then stirred at the same temperature for 30 minutes. The first mixture was added dropwise to the second mixture at 0°C. The mixture was stirred at 10°C for 2 hours. LCMS indicated the reaction was complete. The reaction mixture was quenched with 2N NH4Cl (10 mL) and diluted with EtOAc (200 mL). The organic layer was washed with water (50 mL), brine (50 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified using reverse-phase chromatography (SepaFlash® C18, BOSTON, 0-100% CH3CN / 10 mM NH4HCO3 in water) to afford isopropyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (1 g, 1.368 mmol, 13.37% yield) as a white solid. LCMS (M+H) = 639.0; retention time (10mM NH4HCO3) = 1.67 min.

[0195] Step 4: (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((((S)-1-isopropoxy-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl icosanoate To a solution of icosanoic acid (323 mg, 1.034 mmol) in DCM (40 mL) were added EDC (901 mg, 4.70 mmol) and DMAP (574 mg, 4.70 mmol), and the resulting mixture was stirred at room temperature for 0.5 h. Then, isopropyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (600 mg, 0.940 mmol) was added, and the resulting mixture was stirred at room temperature overnight. LCMS showed the reaction was complete. The reaction mixture was diluted with DCM (200 mL), and the organic layer was separated, washed with water (40 mL), brine (40 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified using reverse-phase chromatography (SepaFlash® C18, BOSTON, 0-100% CHCN / 10 mM NHHCO in water and 100% THF) to give (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((((S)-1-isopropoxy-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl icosanoate (600 mg, 0.643 mmol, 68.41% yield) as a colorless oil. The diastereomers were separated by preparative SFC (instrument: SFC-80 (Thar, Waters), column: IG 20 × 250 mm, 10 μm (Daicel), column temperature: 40 °C, mobile phase: CO / EtOH (1.0% ammonia / methanol) = 35 / 65, flow rate: 80 g / min, back pressure: 100 bar, detection wavelength: 260 nm, cycle time: 15 min) to give the first-eluting isomer (Example 6A, RT1: 1.57 min, 135 mg, 0.145 mmol, yield: 15.40%) as a light gray solid, followed by the second-eluting isomer (Example 6B, RT2: 2.53 min, 115 mg, 0.123 mmol, yield: 13.12%) as a brown oil. First eluting isomer: LCMS (M+H) = 933.5; retention time (0.05% TFA) = 4.05 min. HPLC: retention time (10 mM NH4HCO3) = 14.89 min.1 H NMR (400MHz, CDCl3) δ 8.06 (s, 1H), 7.22-7.10 (m, 7H), 7.08-7.04 (m, 2H), 6.43 (t, J = 6.8 Hz, 1H), 6.04 (brs, 2H), 5.57 (dd, J = 6.8, 4.0 Hz, 1H), 4.94 (quint, J = 6.4 Hz, 1H), 4.29 (dd, J = 10.8, 6.8 Hz, 1H), 4.22-4.15 (m, 1H), 4.09 (dd, J = 10.8, 5.6 Hz, 1H), 3.60 (t, J = 10.8 Hz, 1H), 2.96 (d, J = 6.0 Hz, 2H), 2.61 (s, 1H), 2.61-2.50 (m, 2H), 2.38 (t, J = 7.6 Hz, 2H), 1.65 (quint, J = 7.2 Hz, 2H), 1.30-1.22 (m, 32H), 1.17 (d, J = 6.0 Hz, 3H), 1.13 (d, J = 6.0 Hz, 3H), 0.88 (t, J = 6.8 Hz, 3H). Chapter 2: Dissolved heterogeneous body: LCMS (M+H) = 934.4; retention time (0.05% TFA) = 4.03 minutes. HPLC: retention time (10mM NH4HCO3) = 15.31 minutes. 1H NMR (400MHz, CDCl3) δ 7.99 (s, 1H), 7.24-7.20 (m, 5H), 7.16 (d, J = 7.6 Hz, 2H), 7.12 (d, J = 7.6 Hz, 3H), 6.40 (t, J = 6.8 Hz, 1H), 5.91 (brs, 2H), 5.57 (dd, J = 6.8, 4.0 Hz, 1H), 4.94 (quint, J = 6.4 Hz, 1H), 4.30-4.22 (m, 2H), 4.13 (dd, J = 10.8, 5.6 Hz, 1H), 3.60 (t, J = 10.8 Hz, 1H), 2.99 (d, J = 6.4 Hz, 2H), 2.69-2.55 (m, 3H), 2.40 (t, J = 7.6 Hz, 2H), 1.72-1.68 (m, 2H), 1.28-1.22 (m, 32H), 1.14 (dd, J = 8.0, 6.4 Hz, 6H), 0.88 (t, J = 6.8 Hz, 3H).

[0196] Example 7

[0197] [ka]

[0198] Step 1: Isopropyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate To a cooled (ice-water bath) solution of isopropyl L-alaninate hydrochloride (2.095 g, 12.50 mmol) and triethylamine (3.48 mL, 25.00 mmol) in DCM (60 mL), a solution of phenyl phosphorodichloridate (2.637 g, 12.50 mmol) in DCM (1 mL) was added dropwise under a nitrogen atmosphere, and the reaction mixture was stirred at the same temperature for 1 hour. To a cooled (ice-water bath) solution of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (1466 mg, 5 mmol) in THF (60 mL) and pyridine (20 mL), tert-butylmagnesium chloride (12.50 mL, 12.50 mmol) was added dropwise under a nitrogen atmosphere, and the mixture was stirred at room temperature for 30 minutes. The first mixture was added dropwise to the second mixture at room temperature and stirred for 2 hours. LCMS showed the reaction was complete. The reaction mixture was filtered, the filtrate was concentrated, and the residue was then diluted with EtOAc (200 mL). The organic layer was washed with water (40 mL), 1N HCl (40 mL), brine (40 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified using silica gel chromatography (DCM / MeOH=20 / 1) to give isopropyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (510 mg, 0.907 mmol, 18.13% yield) as a white solid. LCMS (M+H)=563.0; retention time (0.1%TFA)=1.48, 1.49 min.

[0199] Step 2: (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl icosanoate To a solution of icosanoic acid (306 mg, 0.978 mmol) in DCM (20 mL) were added DMAP (543 mg, 4.44 mmol) and EDC (852 mg, 4.44 mmol), and the resulting mixture was stirred at room temperature for 0.5 h. Then, isopropyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (500 mg, 0.889 mmol) was added, and the resulting mixture was stirred at room temperature overnight. LCMS showed the reaction was complete. The reaction mixture was diluted with DCM (100 mL), washed with water (30 mL), washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified using reverse-phase chromatography (SepaFlash® C18, BOSTON, 0-100% 10 mM NH4HCO3 / MeOH, 98%) to afford (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl icosanoate (450 mg, 0.526 mmol, 59.13% yield) as a white solid. The diastereomers were separated by preparative SFC (instrument: SFC-80 (Thar, Waters); column: AD 20 × 250 mm, 10 μm (Daicel); column temperature: 40 °C; mobile phase: CO / MeOH (1.0% ammonia / methanol) = 45 / 55; flow rate: 80 g / min; back pressure: 100 bar; detection wavelength: 260 nm; cycle time: 8.7 min) to give the first-eluting isomer (Example 7A, RT1: 1.24 min, 160 mg, 0.187 mmol, yield: 21.00%) as a white solid, followed by the second-eluting isomer (Example 7B, RT2: 2.24 min, 211 mg, 0.243 mmol, yield: 27.4%) as a white semi-solid. First eluting isomer: LCMS (M+H) = 857.3; retention time (0.05% TFA) = 3.76 min. HPLC retention time (10 mM NH4HCO3) = 13.23 min. 1H NMR (400MHz, CDCl3) δ 8.04 (s, 1H), 7.32-7.28 (m, 2H), 7.25-7.20 (m, 2H), 7.13 (t, J = 7.2 Hz, 1H), 6.45 (t, J = 6.8 Hz, 1H), 6.11 (brs, 2H), 5.67 (dd, J = 6.8, 4.0 Hz, 1H), 4.98 (quint, J = 6.4 Hz, 1H), 4.42 (dd, J = 6.0, 3.2 Hz, 2H), 4.02-3.93 (m, 1H), 3.86-3.80 (m, 1H), 2.75 (quint, J = 7.2 Hz, 1H), 2.64 (s, 1H), 2.57 (ddd, J = 14.0, 6.0, 4.0 Hz, 1H), 2.39 (t, J = 7.6 Hz, 2H), 1.66 (quint, J = 7.6 Hz, 2H), 1.30 (t, J = 7.2 Hz, 3H), 1.32-1.22 (m, 32H), 1.20 (dd, J = 8.0, 6.4 Hz, 6H), 0.88 (t, J = 6.8 Hz, 3H). Chapter 2: Dissolved heterogeneous body: LCMS (M+H) = 857.4; retention time (0.05% TFA) = 3.79 minutes. HPLC retention time (10mM NH4HCO3) = 13.36 minutes. 1H NMR (400MHz, CDCl3) δ 8.09 (s, 1H), 7.32-7.28 (m, 2H), 7.20 (d, J = 8.8 Hz, 2H), 7.13 (t, J = 7.2 Hz, 1H), 6.43 (t, J = 6.8 Hz, 1H), 6.02 (brs, 2H), 5.65 (dd, J = 6.8, 4.0 Hz, 1H), 5.01 (quint, J = 6.4 Hz, 1H), 4.43 (dd, J = 11.2, 6.8 Hz, 1H), 4.34 (dd, J = 11.2, 6.0 Hz, 1H), 4.01 (dd, J = 16.0, 8.8 Hz, 1H), 3.80 (t, J = 10.4 Hz, 1H), 2.71 (quint, J = 6.8 Hz, 1H), 2.67-2.60 (m, 2H), 2.40 (t, J = 7.2 Hz, 2H), 1.66 (quint, J = 7.6 Hz, 2H), 1.36 (d, J = 6.8 Hz, 3H), 1.30-1.24 (m, 32H), 1.22 (d, J = 6.4 Hz, 6H), 0.88 (t, J = 6.8 Hz, 3H).

[0200] Example 8

[0201]

change

[0202] Project 1:2 - Tert-Tert-Tert-L-Arm To a mixture of 2-ethylbutan-1-ol (5.11 g, 50.0 mmol), (tert-butoxycarbonyl)-L-alanine (9.46 g, 50 mmol), and HATU (26.6 g, 70.0 mmol) in DCM (200 mL) was added 1H-imidazole (10.21 g, 150 mmol) and DIPEA (26.2 mL, 150 mmol). The reaction mixture was stirred at 25 °C overnight. LCMS showed the reaction was complete. The reaction mixture was quenched with water (100 mL) and extracted with DCM (3 × 150 mL). The combined organic layers were washed with brine (100 mL), dried over NaSO, filtered, and concentrated to give 2-ethylbutyl (tert-butoxycarbonyl)-L-alaninate (13.67 g, 50.0 mmol, 100% yield) as a colorless oil, which was used in the next step without further purification: LCMS (M+Na) = 296.1; retention time (0.1% TFA) = 2.11 min.

[0203] Step 2: 2-Ethylbutyl L-alaninate To a solution of 2-ethylbutyl (tert-butoxycarbonyl)-L-alaninate (13.67 g, 50 mmol) in DCM (40 mL) was added TFA (20 mL, 260 mmol). The resulting reaction mixture was stirred at 25 °C for 3 h. LCMS showed the reaction was complete. The mixture was concentrated, and the residue was diluted with NaHCO (50 mL) and extracted with DCM (3 × 100 mL). The combined organic layers were washed with brine (80 mL), dried over NaSO, filtered, and concentrated. The residue was purified using silica gel chromatography (DCM / MeOH = 10 / 1) to give 2-ethylbutyl L-alaninate (8 g, 46.2 mmol, 92% yield) as a colorless oil. LCMS (M+H) = 174.1; retention time (0.1% TFA) = 1.17 min.

[0204] Step 3: 2-Ethylbutyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate To a cooled (ice-water bath) solution of 2-ethylbutyl L-alaninate (1.182 g, 6.82 mmol) and triethylamine (1.188 mL, 8.52 mmol) in DCM (42 mL), a solution of phenyl phosphorodichloridate (1439 mg, 6.82 mmol) in DCM (1 mL) was added dropwise under a nitrogen atmosphere, and the reaction mixture was stirred at the same temperature for 1 hour. To a solution of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (1 g, 3.41 mmol) in THF (28 mL) and pyridine (14 mL), tert-butylmagnesium chloride (8.52 mL, 8.52 mmol) was added dropwise under a nitrogen atmosphere, and the mixture was stirred at the same temperature for 30 minutes. The first prepared mixture was then added dropwise to the second prepared mixture at 0° C. The mixture was stirred at 10° C. for 2 hours. LCMS showed the reaction was complete. The reaction mixture was filtered and concentrated. The residue was diluted with EtOAc (200 mL), washed with water (50 mL), washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified using preparative HPLC to give 2-ethylbutyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (520 mg, 0.774 mmol, 22.70% yield) as a white solid. LCMS (M+H) = 605.3; retention time (0.05% TFA) = 1.65 min.

[0205] Step 4: (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((((S)-1-(2-ethylbutoxy)-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl icosanoate To a mixture of 2-ethylbutyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (520 mg, 0.860 mmol) and EDC (824 mg, 4.30 mmol) in DCM (10 mL) was added icosanoic acid (323 mg, 1.032 mmol). The resulting reaction mixture was stirred at room temperature overnight and concentrated to give the crude product. The residue was purified using flash chromatography (silica gel, 120 g, 0-100% DCM / MeOH) to afford (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((((S)-1-(2-ethylbutoxy)-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl icosanoate (450 mg, 0.450 mmol, 52.4% yield) as a white semi-solid. LCMS (M+H) = 900.1; retention time (0.05% TFA) = 4.32 min. The diastereomers were separated by preparative SFC (instrument: SFC-80 (Thar, Waters); column: IG 20 × 250 mm, 10 μm (Daicel); column temperature: 40 °C; mobile phase: CO / EtOH (1.0% ammonia / methanol) = 35 / 65; flow rate: 80 g / min; back pressure: 100 bar; detection wavelength: 260 nm; cycle time: 6 min) to give the first-eluting isomer (Example 8A, RT: 1.16 min, 142 mg, 0.155 mmol, yield: 43.5%) as a light gray semi-solid, followed by the second-eluting isomer (Example 8B, RT: 1.88 min, 105 mg, 0.117 mmol, yield: 32.8%) as a light gray semi-solid. First eluting isomer: LCMS (M+H) = 900.5; retention time (0.05% TFA) = 4.29 min. HPLC retention time (10 mM NH4HCO3) = 16.18 min. 1H NMR (400MHz, CDCl3) δ 8.05 (s, 1H), 7.32-7.27 (m, 2H), 7.22 (d, J = 8.0 Hz, 2H), 7.13 (t, J = 7.6 Hz, 1H), 6.45 (t, J = 6.8 Hz, 1H), 6.13 (brs, 2H), 5.67 (dd, J = 6.8, 4.0 Hz, 1H), 4.42-4.39 (m, 2H), 4.09-3.98 (m, 3H), 3.85 (t, J = 10.8 Hz, 1H), 2.75 (quint, J = 6.8 Hz, 1H), 2.64 (s, 1H), 2.57 (ddd, J = 13.6, 6.0, 4.0 Hz, 1H), 2.39 (t, J = 7.6 Hz, 2H), 1.66 (quint, J = 7.2 Hz, 2H), 1.49 (quint, J = 6.0 Hz, 1H), 1.36-1.24 (m, 39H), 0.90-0.83 (m, 9H). The isomer dissolved in the second part: LCMS (M+H) = 899.5; retention time (0.05% TFA) = 4.34 minutes. HPLC retention time (10 mM NH4HCO3) = 16.53 minutes. 1H NMR (400MHz, CDCl3) δ 8.05 (s, 1H), 7.32-7.27 (m, 2H), 7.23-7.18 (m, 2H), 7.13 (t, J = 7.6 Hz, 1H), 6.43 (t, J = 6.8 Hz, 1H), 6.03 (brs, 2H), 5.65 (dd, J = 6.8, 4.0 Hz, 1H), 4.43 (dd, J = 11.2, 6.4 Hz, 1H), 4.35 (dd, J = 11.2, 6.0 Hz, 1H), 4.11-4.05 (m, 2H), 3.99 (dd, J = 10.8, 5.6 Hz, 1H), 3.84 (dd, J = 11.2, 9.6 Hz, 1H), 2.72 (quint, J = 7.2 Hz, 1H), 2.66-2.59 (m, 2H), 2.40 (t, J = 7.6 Hz, 2H), 1.66 (quint, J = 7.6 Hz, 2H), 1.50 (quint, J = 6.0 Hz, 1H), 1.39 (d, J = 7.2 Hz, 3H), 1.35-1.25 (m, 36H), 0.90-0.85 (m, 9H).

[0206] Example 9

[0207]

change

[0208] Project 1:2-Teret-Tower-L-Tower To a mixture of 2-ethylbutan-1-ol (5.11 g, 50.0 mmol), (tert-butoxycarbonyl)-L-phenylalanine (13.27 g, 50 mmol), and HATU (26.6 g, 70.0 mmol) in DCM (200 mL) was added 1H-imidazole (10.21 g, 150 mmol) and DIPEA (26.2 mL, 150 mmol). The resulting reaction mixture was stirred at 25 °C overnight. LCMS showed the reaction was complete. The reaction was diluted with water (100 mL), the organic layer was separated, and the aqueous layer was extracted with DCM (2 × 150 mL). The combined organic layers were washed with brine (100 mL), dried over NaSO, filtered, and concentrated to give 2-ethylbutyl (tert-butoxycarbonyl)-L-phenylalaninate (17.45 g, 49.9 mmol, 100% yield) as a colorless oil, which was used in the next step without further purification: LCMS (M+Na) = 372.0; retention time (0.1% TFA) = 2.11 min.

[0209] Step 2: 2-Ethylbutyl L-phenylalaninate To a solution of 2-ethylbutyl (tert-butoxycarbonyl)-L-phenylalaninate (17.45 g, 49.9 mmol) in DCM (40 mL) was added 2,2,2-trifluoroacetic acid (20 mL, 260 mmol), and the mixture was stirred at 25 °C for 3 h. LCMS indicated the reaction was complete. The mixture was concentrated, and the residue was purified using silica gel chromatography (DCM / MeOH = 10 / 1) to give 2-ethylbutyl L-phenylalaninate (9.5 g, 38.1 mmol, 76.35% yield) as a colorless oil. LCMS (M+H) = 250.2; retention time (0.01% TFA) = 1.46 min.

[0210] Step 3: 2-Ethylbutyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate A solution of phenyl phosphorodichloridate (2.55 mL, 17.05 mmol) in DCM (5 mL) was added dropwise to a solution of 2-ethylbutyl L-phenylalaninate (4.25 g, 17.05 mmol) and triethylamine (2.376 mL, 17.05 mmol) in DCM (50 mL) under a nitrogen atmosphere at 0° C., and the reaction mixture was stirred at 0° C. for 1 hour. To a solution of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (2 g, 6.82 mmol) in THF (20 mL) and pyridine (10 mL), tert-butylmagnesium chloride (17.05 mL, 17.05 mmol) was added dropwise under a nitrogen atmosphere at 0° C. The mixture was then stirred at the same temperature for 30 minutes. The first mixture was added dropwise to the second mixture at 0°C and stirred at 10°C for 2 hours. LCMS indicated the reaction was complete. The reaction mixture was filtered and concentrated. The residue was diluted with DCM (200 mL), washed with water (40 mL), dried over NaSO, filtered, and concentrated. The residue was purified using reverse-phase chromatography (SepaFlash® C18, BOSTON, 0-100% CHCN / 10 mM NHHCO in water) to afford 2-ethylbutyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (850 mg, 1.073 mmol, 15.73% yield) as a white solid. LCMS (M+H) = 681.0; retention time (10mM NH4HCO3) = 1.83 min.

[0211] Step 4: (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((((S)-1-(2-ethylbutoxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl icosanoate To a solution of icosanoic acid (303 mg, 0.970 mmol) in DCM (50 mL) were added DMAP (538 mg, 4.41 mmol) and EDC (845 mg, 4.41 mmol), and the resulting mixture was stirred at room temperature for 0.5 h. Then, 2-ethylbutyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (600 mg, 0.881 mmol) was added, and the resulting mixture was stirred at room temperature overnight. LCMS showed that the reaction was complete. The reaction mixture was diluted with DCM (100 mL), washed with water (30 mL), brine (30 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified using reverse-phase chromatography (SepaFlash® C18, BOSTON, 0-100% MeOH / 10 mM NH4HCO3 in water) to give (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((((S)-1-(2-ethylbutoxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl icosanoate (600 mg, 0.615 mmol, 69.81% yield) as a colorless oil. The diastereomers were separated by preparative SFC (instrument: SFC-80 (Thar, Waters); column: IG 20 × 250 mm, 10 μm (Daicel); column temperature: 40 °C; mobile phase: CO / EtOH (1.0% ammonia / methanol) = 35 / 65; flow rate: 80 g / min; back pressure: 100 bar; detection wavelength: 260 nm; cycle time: 9.6 min) to give the first-eluting isomer (Example 9A, RT: 1.58 min, 130 mg, 0.133 mmol, yield: 15.12%) as a light gray solid, followed by the second-eluting isomer (Example 9B, RT: 2.32 min, 170 mg, 0.172 mmol, yield: 19.50%) as a colorless oil. First eluting isomer: LCMS (M+H) = 976.3; retention time (0.05% TFA) = 4.70 min. HPLC retention time (10 mM NH4HCO3) = 17.22 min. 1H NMR (400MHz, CDCl3) δ 8.01 (s, 1H), 7.25-7.10 (m, 8H), 7.04 (dd, J = 8.0, 2.0 Hz, 2H), 6.43 (t, J = 6.8 Hz, 1H), 6.06 (brs, 2H), 5.59 (dd, J = 6.8, 4.0 Hz, 1H), 4.29 (dd, J = 10.8, 6.4 Hz, 1H), 4.27-4.21 (m, 1H), 4.11 (dd, J = 10.8, 6.0 Hz, 1H), 3.99 (dd, J = 10.8, 5.6 Hz, 1H), 3.92 (dd, J = 10.8, 6.0 Hz, 1H), 3.65 (t, J = 10.8 Hz, 1H), 2.97 (t, J = 6.8 Hz, 2H), 2.65-2.58 (m, 2H), 2.51 (ddd, J = 13.6, 6.0, 3.6 Hz, 1H), 2.38 (t, J = 7.6 Hz, 2H), 1.65 (quint, J = 7.6 Hz, 2H), 1.43 (quint, J = 6.4 Hz, 1H), 1.32-1.22 (m, 36H), 0.88 (t, J = 6.8 Hz, 3H), 0.83 (t, J = 7.6 Hz, 6H). Chapter 2: Dissolved heterogeneous body: LCMS (M+H) = 975.3; retention time (0.05% TFA) = 4.73 minutes. HPLC retention time (10mM NH4HCO3) = 19.12 minutes. 1H NMR (400MHz, CDCl3) δ 7.99 (s, 1H), 7.26-7.21 (m, 4H), 7.20-7.14 (m, 3H), 7.11 (dd, J = 6.8, 1.6 Hz, 3H), 6.40 (t, J = 6.8 Hz, 1H), 6.20 (brs, 2H), 5.57 (dd, J = 6.8, 4.4 Hz, 1H), 4.39-4.29 (m, 1H), 4.24 (dd, J = 10.8, 6.4 Hz, 1H), 4.13 (dd, J = 11.2, 5.6 Hz, 1H), 3.95 (dd, J = 6.0, 2.0 Hz, 2H), 3.74 (t, J = 10.8 Hz, 1H), 3.06-2.95 (m, 2H), 2.67-2.54 (m, 3H), 2.40 (t, J = 7.2 Hz, 2H), 1.67 (quint, J = 7.6 Hz, 2H), 1.42 (quint, J = 6.4 Hz, 1H), 1.32-1.21 (m, 36H), 0.88 (t, J = 6.8 Hz, 3H), 0.83 (t, J = 7.6 Hz, 6H).

[0212] Example 10

[0213]

change

[0214] Project 1: Tert-ブトキシカルボニル-L-フェニルアラニネート To a mixture of hexadecan-1-ol (15 g, 61.9 mmol), (tert-butoxycarbonyl)-L-phenylalanine (16.41 g, 61.9 mmol), 1H-imidazole (12.64 g, 186 mmol), and HATU (35.3 g, 93 mmol) in DCM (200 mL) was added DIPEA (32.4 mL, 186 mmol) and stirred at room temperature for 16 h. TLC showed the reaction was complete. The reaction mixture was washed with water, dried over Na2SO4, filtered, and concentrated to give the crude product. The crude product was purified using flash chromatography (petroleum ether: EtOAc = 20:1) to give hexadecyl (tert-butoxycarbonyl)-L-phenylalaninate (23 g, 47.0 mmol, 76% yield) as a white solid.

[0215] Step 2: Hexadecyl L-phenylalaninate To a solution of hexadecyl (tert-butoxycarbonyl)-L-phenylalaninate (23 g, 47.0 mmol) in DCM (500 mL) was added trifluoroacetic acid (100 mL) and stirred at room temperature for 12 h. TLC indicated the reaction was complete. The reaction mixture was concentrated, and the pH of the residue was adjusted to 8 with NaHCO (aq) and extracted twice with EtOAc (500 mL). The combined organic phases were washed with water (300 mL), brine (300 mL), dried over NaSO, filtered, and concentrated to give the crude product. The crude product was purified using flash chromatography (petroleum ether: EtOAc = 5:1) to give hexadecyl L-phenylalaninate (18 g, 46.2 mmol, 98% yield) as a white solid.

[0216] Step 3: Hexadecyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a cold (ice-water bath) solution of 1-((L-phenylalanyl)oxy)hexadecan-7-ylium (5301 mg, 13.64 mmol) and triethylamine (1.901 mL, 13.64 mmol) in DCM (42 mL) was added dropwise a solution of phenyl phosphorodichloridate (2878 mg, 13.64 mmol) in DCM (1 mL) under a nitrogen atmosphere, and the reaction mixture was then stirred at room temperature for 1 hour. To a cooled (ice-water bath) solution of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (2000 mg, 6.82 mmol) in THF (28 mL) / pyridine (14 mL) was added tert-butylmagnesium chloride (17.05 mL, 17.05 mmol) dropwise under a nitrogen atmosphere, and the mixture was then stirred at room temperature for 30 minutes. The first mixture was added dropwise to the second mixture at room temperature, and the mixture was stirred for 16 hours. LCMS showed the reaction was complete. The reaction mixture was filtered and concentrated. The residue was diluted with EtOAc (150 mL) and washed twice with water (80 mL). The organic phase was dried over Na2SO4, filtered, and concentrated to give the crude product. The crude product was purified using preparative HPLC to give hexadecyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (1.3 g, 1.472 mmol, 21.59% yield) as a white solid. LCMS (M+H) = 821.0; retention time (10 mM NH4HCO3) = 3.162 min.

[0217] Step 4: (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((((S)-1-(hexadecyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl icosanoate To a solution of icosanoic acid (293 mg, 0.938 mmol) in DCM (20 mL), DMAP (521 mg, 4.26 mmol) and EDC (817 mg, 4.26 mmol) were added, and the resulting mixture was stirred at room temperature for 0.5 h. Then, hexadecyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (700 mg, 0.853 mmol) was added, and the resulting mixture was stirred at room temperature overnight. TLC (DCM:MeOH = 10:1, Rf = 0.8) indicated the reaction was complete. The reaction mixture was quenched with water (10 mL) and extracted with DCM (20 mL × 3). The combined organic phase was washed with brine (30 mL), dried over NaSO, and concentrated in vacuo. The crude product was purified using preparative TLC (DCM:CHOH = 10:1, Rf = 0.7) to give (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((((S)-1-(hexadecyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl icosanoate (290 mg, 0.260 mmol, 30.5% yield). The diastereomers were separated by preparative SFC (instrument: SFC-80 (Thar, Waters); column: OZ 20 × 250 mm, 10 μm (Daicel); column temperature: 40 °C; mobile phase: CO / MEOH (1.0% ammonia / methanol) = 40 / 60; flow rate: 80 g / min; back pressure: 100 bar; detection wavelength: 260 nm; cycle time: 9.5 min; sample solution: 290 mg dissolved in 17 mL of methanol; injection volume: 3 mL) to give the first-eluting isomer (Example 10A, RT: 1.88 min, 90 mg, 0.078 mmol, 29.9% yield) as a white solid, followed by the second-eluting isomer (Example 10B, RT: 2.93 min, 96 mg, 0.084 mmol, 32.3% yield) as a white solid. First eluting isomer: HPLC: Retention time (0.1% NH4HCO3) = 38.538 min. 1H NMR (400MHz, CDCl3) δ 8.01 (s, 1H), 7.27 (d, J = 6.7 Hz, 1H), 7.25-7.20 (m, 2H), 7.20 -7.15 (m, 3H), 7.15-7.09 (m, 2H), 7.05-7.01 (m, 2H), 6.42 (t, J = 6.8 Hz, 1H), 6.14 (s, 2H), 5.58 (dd, J = 6.6, 3.8 Hz, 1H), 4.29 (dd, J = 11.0, 6.6 Hz, 1H), 4.26-4.18 (m, 1H), 4.10 (dd, J = 11.0, 5.8 Hz, 1H), 4.01 (t, J = 6.1 Hz, 2H), 3.69 (t, J = 7.0 Hz, 1H), 2.96 (d, J = 6.2 Hz, 2H), 2.61 (s, 1H), 2.57 (dd, J = 12.9, 5.9 Hz, 1H), 2.54-2.47 (m, 1H), 2.38 (t, J = 7.5 Hz, 2H), 1.76 (s, 6H), 1.69-1.60 (m, 2H), 1.24 (d, J = 7.6 Hz, 54H), 0.87 (t, J = 6.7 Hz, 6H). Chapter 2: Dissolution of the opposite body: HPLC: retention time (0.1% NH4HCO3) = 39.401 minutes. 1H NMR (400MHz, CDCl3) δ 8.01 (s, 1H), 7.25-7.18 (m, 5H), 7.15 (d, J = 8.5 Hz, 2H), 7.11 (t, J = 5.7 Hz, 3H), 6.39 (t, J = 6.7 Hz, 1H), 5.99 (s, 2H), 5.59-5.54 (m, 1H), 4.31 (ddd, J = 16.5, 10.2, 6.3 Hz, 1H), 4.25 (dd, J = 11.0, 6.2 Hz, 1H), 4.13 (dd, J = 11.0, 5.7 Hz, 1H), 4.02 (t, J = 6.7 Hz, 2H), 3.63 (t, J = 8.6 Hz, 1H), 3.00 (d, J = 6.4 Hz, 2H), 2.63-2.54 (m, 3H), 2.40 (t, J = 7.6 Hz, 2H), 1.79-1.62 (m, 6H), 1.25 (s, 56H), 0.88 (t, J = 6.8 Hz, 6H).

[0218] Example 11

[0219]

change

[0220] Project 1: Turret To a solution of octadecan-1-ol (32.5 g, 120 mmol), (tert-butoxycarbonyl)-L-phenylalanine (29 g, 109 mmol), and HATU (62.3 g, 164 mmol) in DCM (100 mL) was added 1H-imidazole (22.32 g, 328 mmol) and DIPEA (57.3 mL, 328 mmol). The resulting mixture was stirred at 25 °C overnight. TLC (petroleum ether: EtOAc = 10:1, Rf = 0.5) showed the reaction was complete. The reaction was quenched with water (200 mL) and DCM (100 mL). The organic layer was then separated, and the aqueous layer was extracted with DCM (2 × 50 mL). The combined organic layers were washed with brine (200 mL), dried over Na SO , filtered, and concentrated to give the crude product. The residue was purified using combiflash (330 g silica gel column, hexane:ethyl acetate=20:1). Appropriate fractions containing the desired product were combined and concentrated in vacuo to give octadecyl (tert-butoxycarbonyl)phenylalaninate (32 g, 61.8 mmol, 56.5% yield) as a yellow solid.

[0221] Step 2: Octadecyl phenylalaninate To a solution of octadecyl (tert-butoxycarbonyl)phenylalaninate (32 g, 61.8 mmol) in DCM (400 mL) was added TFA (28.6 mL, 371 mmol). The resulting mixture was stirred at 25 °C for 2 h. TLC (petroleum ether: EtOAc = 10:1, Rf = 0.1) showed the reaction was complete. The reaction was quenched with 1 N NaHCO (200 mL) and DCM (100 mL), and then the organic layer was separated. The aqueous layer was extracted with DCM (2 × 100 mL). The combined organic layers were washed with brine (200 mL), dried over NaSO, and concentrated to give crude octadecyl phenylalaninate (19 g, 45.5 mmol, 73.6% yield), which was used in the next step without purification.

[0222] Step 3: Octadecyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)phenylalaninate To a solution of octadecyl L-phenylalaninate (5.70 g, 13.64 mmol) and triethylamine (1.901 mL, 13.64 mmol) in DCM (80 mL) was added dropwise a solution of phenyl phosphorodichloridate (2.038 mL, 13.64 mmol) in DCM (80 mL) at approximately 5°C under a nitrogen atmosphere. The reaction mixture was then stirred at room temperature for 1 hour. To a solution of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (2 g, 6.82 mmol) in THF (40 mL) and pyridine (20 mL) was added tert-butylmagnesium chloride (17.05 mL, 17.05 mmol) dropwise under a nitrogen atmosphere at 0°C. The mixture was then stirred at room temperature for 30 minutes. The first prepared mixture was added dropwise to the second prepared mixture at room temperature and stirred for 2 hours. LCMS showed the reaction was complete. The mixture was filtered and concentrated to give a yellow solid. The solid was then dissolved in EtOAc (40 mL) and partitioned with HCl (0.05 N, 20 mL). The layers were separated and the aqueous layer was extracted with EtOAc (40 mL). The combined organic phase was washed with brine, dried over Na2SO4, filtered, and concentrated to give the crude product. The residue was purified using reverse-phase chromatography (SepaFlash® C18, BOSTON, 0–100% CH3CN / 10 mM NH4HCO3 in water) to give octadecyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)phenylalaninate (1.2 g, 1.354 mmol, 19.85% yield) as a yellow solid. LCMS (M+H) = 849.4; retention time (0.1% TFA) = 3.568 min.The diastereomers were separated by preparative SFC (instrument: SFC-150 (Waters); column: IH 20 × 250 mm, 10 μm (Daicel); column temperature: 35 °C; mobile phase: CO2 / MeOH (0.2% ammonia / methanol) = 60 / 40; flow rate: 100 g / min; back pressure: 100 bar; detection wavelength: 214 nm; cycle time: 4 min; sample solution: 1,900 mg dissolved in 100 mL methanol; injection volume: 3 mL) to give the first-eluting isomer (Intermediate 11A, RT: 1.76 min, 800 mg, 0.908 mmol, 38.5% yield) and the second-eluting isomer (Intermediate 11B, RT: 3.21 min, 650 mg, 0.766 mmol, 32.5% yield) as white solids. First eluting isomer: LCMS (M+H) = 849.4; Retention time (0.1% TFA) = 2.874 min. HPLC: Retention time (0.1% NH4HCO3): 11.004 min. 1 H NMR (400MHz, CDCl3) δ 7.92 (s, 1H), 7.25-7.17 (m, 5H), 7.14 (d, J = 7.3 Hz, 1H), 7.12-7.08 (m, 4H), 6.31 (dd, J = 7.3, 4.3 Hz, 1H), 6.10 (s, 2H), 4.71 (t, J = 7.6 Hz, 1H), 4.26 (dd, J = 11.2, 7.7 Hz, 2H), 4.20 (dd, J = 11.3, 8.8 Hz, 1H), 4.03 (t, J = 6.7 Hz, 2H), 3.88 (s, 1H), 2.99 (d, J = 6.2 Hz, 2H), 2.76-2.63 (m, 3H), 1.86 (s, 2H), 1.24 (d, J = 9.0 Hz, 30H), 0.88 (t, J = 6.8 Hz, 3H). Second eluting isomer: LCMS (M+H) = 849.5; Retention time (0.1% TFA) = 2.839 min, HPLC: Retention time (0.1% NH4HCO3): 10.931 min. 1H NMR (400MHz, CDCl3) δ 7.78 (s, 1H), 7.30-7.26 (m, 2H), 7.23 (dd, J = 8.4, 4.6 Hz, 3H), 7.15 (d, J = 7.8 Hz, 3H), 7.06 (dd, J = 7.5, 1.7 Hz, 2H), 6.32 (dd, J = 7.4, 4.2 Hz, 1H), 6.24 (s, 2H), 4.64 (t, J = 7.6 Hz, 1H), 4.23-4.15 (m, 2H), 4.02 (t, J = 6.7 Hz, 2H), 3.97 (dd, J = 11.5, 8.4 Hz, 1H), 3.73 (t, J = 10.8 Hz, 1H), 3.04-2.92 (m, 2H), 2.70 (s, 1H), 2.66 (dt, J = 13.7, 7.8 Hz, 1H), 2.57 (ddd, J = 13.6, 7.1, 4.2 Hz, 1H), 1.89 (s, 2H), 1.24 (d, J = 4.9 Hz, 30H), 0.87 (t, J = 6.8 Hz, 3H).

[0223] Step 5: (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((((S)-1-(octadecyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl icosanoate To a solution of icosanoic acid (101 mg, 0.324 mmol) in DCM (10 mL) were added DMAP (180 mg, 1.472 mmol) and EDC (282 mg, 1.472 mmol), and the resulting mixture was stirred at room temperature for 0.5 h. Then, octadecyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (Intermediate 11A, 250 mg, 0.294 mmol) was added, and the resulting mixture was stirred at room temperature overnight. TLC showed that the reaction was complete. The reaction mixture was diluted with DCM (100 mL), washed with water, washed with brine, dried over Na2SO4, filtered, and concentrated. The residue was purified using preparative TLC (DCM / MeOH=10 / 1) to give (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((((S)-1-(octadecyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl icosanoate (Example 11A, 145 mg, 0.127 mmol, 43.1% yield) as a white solid. HPLC: Retention time (10 mM NH4HCO3)=60.25 min. 1H NMR (400MHz, CDCl3) δ 7.99 (s, 1H), 7.26-7.18 (m, 5H), 7.15 (d, J = 8.4 Hz, 2H), 7.13-7.08 (m, 3H), 6.39 (t, J = 6.8 Hz, 1H), 6.16 (brs, 2H), 5.56 (dd, J = 6.4, 4.4 Hz, 1H), 4.36-4.29 (m, 1H), 4.25 (dd, J = 10.8, 6.4 Hz, 1H), 4.13 (dd, J = 10.8, 5.6 Hz, 1H), 4.02 (t, J = 6.8 Hz, 2H), 3.73 (t, J = 10.8 Hz, 1H), 3.02-2.98 (m, 2H), 2.65-2.55 (m, 3H), 2.40 (t, J = 7.2 Hz, 2H), 1.67(quint, J = 6.8 Hz, 2H), 1.52(quint, J = 6.0 Hz, 2H), 1.30-1.22 (m, 62H), 0.90-0.85 (m, 6H).

[0224] To a solution of icosanoic acid (101 mg, 0.324 mmol) in DCM (10 mL) were added DMAP (180 mg, 1.472 mmol) and EDC (282 mg, 1.472 mmol). The resulting mixture was stirred at room temperature for 0.5 h. Then, octadecyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (Intermediate 11B, 250 mg, 0.294 mmol) was added, and the resulting mixture was stirred at room temperature overnight. LCMS showed that the reaction was complete. The reaction mixture was diluted with water (10 mL) and extracted with DCM (15 mL × 3). The combined organic phase was washed with brine (20 mL), dried over Na2SO4, and concentrated in vacuo. The crude product was purified using preparative TLC (DCM:CHOH=10:1, Rf=0.7) to give (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((((S)-1-(octadecyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl icosanoate (Example 11B, 115 mg, 0.101 mmol, 34.2% yield). HPLC: Retention time (0.1% NH4HCO3)=63.275 min. 1H NMR (400MHz, CDCl3) δ 8.03 (s, 1H), 7.27 (d, J = 7.2 Hz, 1H), 7.25-7.20 (m, 2H), 7.18 (d, J = 5.3 Hz, 2H), 7.15 (d, J = 8.2 Hz, 2H), 7.12 (t, J = 7.4 Hz, 1H), 7.06-7.01 (m, 2H), 6.43 (t, J = 6.8 Hz, 1H), 6.13 (s, 2H), 5.58 (dd, J = 6.7, 3.8 Hz, 1H), 4.29 (dd, J = 11.0, 6.6 Hz, 1H), 4.26-4.18 (m, 1H), 4.11 (dd, J = 11.0, 5.8 Hz, 1H), 4.01 (td, J = 6.8, 1.6 Hz, 2H), 3.68 (t, J = 10.7 Hz, 1H), 2.96 (d, J = 6.3 Hz, 2H), 2.64-2.47 (m, 3H), 2.38 (t, J = 7.6 Hz, 2H), 1.82 (s, 2H), 1.69-1.60 (m, 2H), 1.30-1.21 (m, 62H), 0.88 (t, J = 6.8 Hz, 6H).

[0225] Example 12

[0226]

change

[0227] Project 1: Tert-ブトキシカルボニル-L-フェニルアラニネート A mixture of icosan-1-ol (33.8 g, 113 mmol), (tert-butoxycarbonyl)-L-phenylalanine (25 g, 94 mmol), imidazole (19.25 g, 283 mmol), DIPEA (49.4 mL, 283 mmol), and HATU (53.7 g, 141 mmol) in DCM (200 mL) was stirred at 25 °C for 16 h. TLC showed the presence of a new product. Water (200 mL) was added, and the mixture was extracted with DCM (150 mL × 3). The combined organic phases were washed with brine, dried over Na2SO4, filtered, and concentrated. The residue was purified using silica gel chromatography (petroleum ether: EtOAc = 30:1) to give icosyl (tert-butoxycarbonyl)-L-phenylalaninate (20 g, 10.99 mmol, 11.67% yield) as a white solid. 1 H NMR (400MHz, CDCl3) δ 7.3-7.28 (m, 1H), 7.26-7.11 (m, 4H), 4.99 (d, J = 7.4 Hz, 1H), 4.57 (d, J = 7.3 Hz, 1H), 4.12-4.02 (m, 2H), 3.49 (s, 1H), 3.17-2.99 (m, 2H), 1.42 (s, 8H), 1.26 (s, 36H), 0.88 (t, J = 6.8 Hz, 3H).

[0228] Step 2: Eicosyl L-phenylalaninate To a solution of icosyl (tert-butoxycarbonyl)-L-phenylalaninate (30 g, 55.0 mmol) in DCM (200 mL) was added TFA (42.3 mL, 550 mmol) at 0 °C. After 5 min, the mixture was warmed to room temperature and stirred for 2 h. TLC showed complete consumption of the starting material. The pH of the reaction mixture was adjusted to 7-8 with saturated Na2CO3 and extracted with DCM (300 mL × 3). The combined organic phases were washed with brine, dried over Na2SO4, filtered, and concentrated to give a residue. The residue was purified using silica gel chromatography (DCM:MeOH = 20:1) to give icosyl L-phenylalaninate (25 g, 50.5 mmol, 92% yield). 1H NMR (400MHz, DMSO) δ 7.34-7.08 (m, 5H), 4.00-3.88 (m, 2H), 3.55 (t, J = 6.8 Hz, 1H), 2.92- 2.73 (m, 2H), 1.91 (s, 2H), 1.52-1.39 (m, 2H), 1.23 (s, 35H), 0.85 (t, J = 6.8 Hz, 3H).

[0229] Step 3: Icosyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a cold (ice-water bath) solution of icosyl L-phenylalaninate (7.60 g, 17.05 mmol) and triethylamine (2.376 mL, 17.05 mmol) in DCM (48 mL), a solution of phenyl phosphorodichloridate (3.60 g, 17.05 mmol) in DCM (1 mL) was added dropwise under a nitrogen atmosphere in ice water, and the reaction mixture was stirred at room temperature for 1 hour. To a solution of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (2 g, 6.82 mmol) in THF (24 mL) and pyridine (12 mL) was added tert-butylmagnesium chloride (17.05 mL, 17.05 mmol) under a nitrogen atmosphere at 0 °C, and the mixture was warmed to room temperature and stirred for 30 minutes. The first mixture was added dropwise to the second mixture at room temperature and stirred for 2 hours. LCMS indicated the reaction was complete. The reaction mixture was filtered and concentrated, and then the residue was diluted with DCM. Water was added to the mixture, and the mixture was extracted. The organic layer was then washed three times with 0.5 N HCl and brine. The organic phase was dried over Na2SO4, filtered, and concentrated. The residue was purified using flash chromatography (0-5% MeOH / DCM) to give icosyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (800 mg, 0.867 mmol, 12.71% yield). LCMS (M+H) = 877.7; retention time (0.1% TFA) = 3.82 min.

[0230] Step 4: (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((((S)-1-(icosyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl icosanoate To a solution of icosyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (1200 mg, 1.368 mmol) in DCM (10 mL) was added DMAP (167 mg, 1.368 mmol), EDC (1311 mg, 6.84 mmol), DIEA (0.717 mL, 4.10 mmol), and icosanoic acid (428 mg, 1.368 mmol). The resulting mixture was stirred at room temperature overnight. TLC showed the presence of a new compound. The reaction mixture was diluted with water and extracted with DCM (40 mL × 3). The combined organic phase was washed with brine, dried over Na2SO4, filtered, and concentrated to give a residue. The residue was purified using preparative TLC (DCM:MeOH=10:1) to give (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((((S)-1-(icosyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl icosanoate (800 mg, 0.628 mmol, 45.9% yield). The diastereomers were separated by preparative SFC (instrument: SFC-150 (Waters); column: OZ 20 × 250 mm, 10 μm (Daicel); column temperature: 35 °C; mobile phase: CO / MeOH (0.2% ammonia / methanol) = 50 / 50; flow rate: 120 g / min; back pressure: 100 bar; detection wavelength: 214 nm; cycle time: 3 min) to give the first-eluting isomer (Example 12A, 160 mg, 0.137 mmol, 26.7% yield) as an off-white solid, followed by the second-eluting isomer (Example 12B, 150 mg, 0.127 mmol, 24.78% yield) as an off-white solid. First eluting isomer: 1H NMR (400MHz, CDCl3) δ 8.00 (s, 1H), 7.24-6.96 (m, 10H), 6.43 (t, J = 6.8 Hz, 1H), 5.58 (dd, J = 6.8, 3.7 Hz, 1H), 4.26 (ddd, J = 25.6, 13.8, 6.4 Hz, 2H), 4.11 (dd, J = 10.9, 5.8 Hz, 1H), 4.09-3.95 (m, 2H), 3.58 (t, J = 10.8 Hz, 1H), 2.96 (d, J = 6.1 Hz, 2H), 2.66-2.47 (m, 3H), 2.38 (t, J = 7.6 Hz, 2H), 1.53 (s, 4H), 1.36-1.17 (m, 68H), 0.88 (t, J = 6.8 Hz, 6H). Chapter 2: Dissolving the heterosexual body: 1 H NMR (400MHz, CDCl3) δ 8.00 (s, 1H), 7.24-6.95 (m, 10H), 6.41 (t, J = 6.8 Hz, 1H), 5.58 (dd, J = 6.8, 3.7 Hz, 1H), 4.26 (ddd, J = 25.6, 13.8, 6.4 Hz, 2H), 4.11 (dd, J = 10.9, 5.8 Hz, 1H), 4.09-3.95 (m, 2H), 3.58 (t, J = 10.8 Hz, 1H), 2.96 (d, J = 6.1 Hz, 2H), 2.66-2.47 (m, 3H), 2.38 (t, J = 7.6 Hz, 2H), 1.53 (s, 4H), 1.36-1.17 (m, 68H), 0.89 (t, J = 6.8 Hz, 6H).

[0231] Example 13

[0232]

change

[0233] Process 1: Tert-Factory-L-Factory To a mixture of (tert-butoxycarbonyl)-L-phenylalanine (10 g, 37.7 mmol), HOBt (7.64 g, 56.5 mmol), and EDC (8.67 g, 45.2 mmol) in DCM (150 mL) was added DIPEA (19.75 mL, 113 mmol) at 0 °C. After 30 min, decan-1-ol (5.97 g, 37.7 mmol) was added. The resulting reaction mixture was stirred at 25 °C for 4 h. LCMS showed the reaction was complete. The reaction was then diluted with water (100 mL), the organic layer was separated, and the aqueous layer was extracted with DCM (200 mL × 2). The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated. The residue was purified using silica gel chromatography (petroleum ether: EtOAc = 10:1) to give decyl (tert-butoxycarbonyl)-L-phenylalaninate (13 g, 32.1 mmol, 85% yield) as a colorless oil. LCMS: (M + Na) = 428.1; retention time (0.1% TFA) = 2.69 min.

[0234] Step 2: Decyl L-phenylalaninate To a solution of decyl (tert-butoxycarbonyl)-L-phenylalaninate (13 g, 32.1 mmol) in DCM (100 mL) was added TFA (19.76 mL, 256 mmol) at 0 °C. The resulting reaction mixture was stirred at 25 °C for 16 h. LCMS showed the reaction was complete. The reaction mixture was concentrated, and the residue was diluted with DCM (100 mL) and neutralized with NaHCO (50 mL). The organic layer was separated, and the aqueous layer was extracted with DCM (2 × 100 mL). The combined organic layers were washed with brine (80 mL), dried over NaSO, filtered, and concentrated. The residue was purified using silica gel chromatography (petroleum ether: EtOAc = 2:1) to afford decyl L-phenylalaninate (8.2 g, 26.8 mmol, 84% yield) as a yellow oil. LCMS (M+H)=306.1; Retention time (0.1%TFA)=1.74 min.

[0235] Step 3: Decyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a stirred mixture of decyl L-phenylalaninate (4.17 g, 13.64 mmol) and triethylamine (1.380 g, 13.64 mmol) in DCM (60 mL), a solution of phenyl phosphorodichloridate (2.88 g, 13.64 mmol) in DCM (10 mL) was added dropwise at approximately 0° C., and the mixture was stirred at 0° C. for 1 h. To a stirred solution of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (2 g, 6.82 mmol) in THF (40.00 mL) and pyridine (20.0 mL), tert-butylmagnesium chloride (1.993 g, 17.05 mmol) was added dropwise under a nitrogen atmosphere at 0° C., and the mixture was stirred at the same temperature for 30 min. The first mixture was added dropwise to the second mixture at 0°C and stirred at 0°C for 2 hours. LCMS showed the reaction was complete. The reaction mixture was filtered and concentrated. The residue was diluted with EtOAc (600 mL) and washed with water (100 mL × 2). The organic layer was dried over NaSO and concentrated. The residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 0-100% CHCN / 10 mM NHHCO aqueous solution) to give decyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (900 mg) as a white solid.The diastereomers were separated by preparative SFC (instrument: SFC-80 (Thar, Waters), column: AS 20 × 250 mm, 10 μm (Daicel), column temperature: 40 °C, mobile phase: CO2 / MeOH (1.0% The separation was carried out using a 60 / 40 ammonia / methanol (flow rate: 80 g / min), 100 bar back pressure, 260 nm detection wavelength, 10 min cycle time, 900 mg dissolved in 30 mL methanol, 2 mL injection volume, to give the first eluting isomer (Intermediate 13A, 160 mg, 0.209 mmol, 3.06% yield) as a white semi-solid (LCMS (M+H) = 737.2; retention time (0.05% TFA) = 1.99 min), followed by the second eluting isomer (Intermediate 13B, 300 mg, 0.407 mmol, 5.97% yield) as a white solid (LCMS (M+H) = 737.2; retention time (0.05% TFA) = 1.98 min).

[0236] Step 4: (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((((S)-1-(decyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl decanoate To a stirred solution of decanoic acid (38.6 mg, 0.224 mmol) in DCM (10 mL) was added 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (195 mg, 1.018 mmol) and N,N-dimethylpyridin-4-amine (124 mg, 1.018 mmol), and the resulting mixture was stirred at room temperature for 0.5 h. Then, decyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (Intermediate 13A, 150 mg, 0.204 mmol) was added, and the resulting mixture was stirred at room temperature overnight. TLC indicated the reaction was complete. The reaction mixture was partitioned between DCM (100 mL) and water (30 mL). The organic layer was then washed with brine, dried over NaSO, filtered, and concentrated. The residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 0-100% CHOH / 10 mM NHHCO aqueous solution) to afford (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((((S)-1-(decyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl decanoate (114 mg, 0.125 mmol, 61.6% yield) as a colorless oil. LCMS (M+H) = 891; retention time (0.05% TFA) = 3.29 min. HPLC retention time (10mM NH4HCO3) = 11.90 minutes. 1H NMR (400MHz, CDCl3) δ 8.01 (s, 1H), 7.27-7.22 (m, 4H), 7.21-7.15 (m, 3H), 7.14-7.11 (m, 3H), 6.42 (t, J = 6.8 Hz, 1H), 6.00 (brs, 2H), 5.59 (dd, J = 6.8, 4.0 Hz, 1H), 4.38-4.31 (m, 1H), 4.27 (dd, J = 10.8, 6.4 Hz, 1H), 4.15 (dd, J = 11.2, 6.0 Hz, 1H), 4.04 (t, J = 6.8 Hz, 2H), 3.66 (t, J = 6.8 Hz, 1H), 3.02 (d, J = 6.4 Hz, 2H), 2.70-2.57 (m, 3H), 2.42 (t, J = 7.6 Hz, 2H), 1.69 (quint, J = 7.2 Hz, 2H), 1.54 (quint, J = 7.2 Hz, 2H), 1.35-1.23 (m, 26H), 0.93-0.88 (m, 6H).

[0237] To a stirred solution of decanoic acid (51.4 mg, 0.299 mmol) in DCM (10 mL) was added 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (260 mg, 1.357 mmol) and N,N-dimethylpyridin-4-amine (166 mg, 1.357 mmol), and the resulting mixture was stirred at room temperature for 0.5 h. Then, decyl ((((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (Intermediate 13A, 200 mg, 0.271 mmol) was added, and the resulting mixture was stirred at room temperature overnight. TLC indicated the reaction was complete. The reaction mixture was partitioned between DCM (100 mL) and water (30 mL). The organic layer was then washed with brine, dried over NaSO, filtered, and concentrated. The residue was purified using reverse-phase chromatography (SepaFlash® C18, BOSTON, 0-100% CHOH / 10 mM NHHCO in water) to afford (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((((S)-1-(decyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl decanoate (100 mg, 0.110 mmol, 40.6% yield) as a white solid. LCMS (M+H) = 891; retention time (0.05% TFA) = 3.26 min. HPLC retention time (10mM NH4HCO3) = 11.81 minutes. 1H NMR (400MHz, CDCl3) δ 8.01 (s, 1H), 7.25-7.10 (m, 8H), 7.06-7.02 (m, 2H), 6.45-6.41 (m, 1H), 6.20 (brs, 2H), 5.58 (dd, J = 6.8, 3.6 Hz, 1H), 4.29 (dd, J = 11.2, 6.8 Hz, 1H), 4.26-4.19 (m, 1H), 4.10 (dd, J = 11.2, 6.0 Hz, 1H), 4.01 (dt, J = 6.8, 2.0 Hz, 2H), 3.73 (t, J = 10.8 Hz, 1H), 2.96 (d, J = 6.4 Hz, 2H), 2.61 (s, 1H), 2.58 (t, J = 7.2 Hz, 1H), 2.51 (ddd, J = 14.0, 6.4, 4.0 Hz, 1H), 2.38 (t, J = 7.6 Hz, 2H), 1.65 (quint, J = 7.2 Hz, 2H), 1.53 (quint, J = 7.2 Hz, 2H), 1.30-1.23 (m, 26H), 0.91-0.87 (m, 6H).

[0238] Example 14

[0239]

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[0240] Project 1: Tert-ブトキシカルボニル-L-フェニルアラニネート To a stirred mixture of EDC (17.34 g, 90 mmol), HOBt (11.54 g, 75 mmol), and (tert-butoxycarbonyl)-L-phenylalanine (20.0 g, 75 mmol) in DCM (100 mL) was added DIPEA (39.5 mL, 226 mmol) at −10° C. After 30 min, a solution of octan-1-ol (11.78 g, 90 mmol) in DCM (100 mL) was added to the above solution and stirred at 25° C. for 16 h. TLC showed the reaction was complete. The reaction mixture was diluted with water (100 mL), the organic layer was separated, and the aqueous layer was extracted with DCM (80 mL × 3). The combined organic phase was washed with brine (80 mL), dried over Na2SO4, and concentrated. The residue was purified using silica gel chromatography (petroleum ether: EtOAc = 10:1) to give octyl (tert-butoxycarbonyl)-L-phenylalaninate (18.0 g, 45.3 mmol, 60.0% yield) as a white solid. LCMS (M + H) = 377.7; retention time (0.1% TFA) = 2.423 min.

[0241] Step 2: Octyl L-phenylalaninate A mixture of TFA (80 mL, 1038 mmol) and octyl (tert-butoxycarbonyl)-L-phenylalaninate (18.0 g, 47.7 mmol) in DCM (200 mL) was stirred at 25 °C overnight. TLC showed the reaction was complete. The reaction mixture was concentrated, and the pH of the residue was adjusted to 8–9 with NaHCO (1 N) and extracted with DCM (500 mL × 3). The combined organic phase was washed with brine, dried over NaSO, and concentrated. The residue was purified using silica gel chromatography (DCM:MeOH = 10:1) to give octyl L-phenylalaninate (12 g, 43.3 mmol, 91% yield) as a white solid. 1H NMR (400MHz, CDCl3) δ 7.33 - 7.27 (m, 2H), 7.26 - 7.16 (m, 3H), 4.09 (t, J = 6.7 Hz, 2H), 3.72 (dd, J = 7.8, 5.4 Hz, 1H), 3.08 (dd, J = 13.5, 5.4 Hz, 1H), 2.87 (dd, J = 13.5, 7.8 Hz, 1H), 1.64 - 1.57 (m, 2H), 1.28 (s, 10H), 0.89 (t, J = 6.9 Hz, 3H).

[0242] Step 3: Octyl ((perfluorophenoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a stirred solution of octyl L-phenylalaninate (5.00 g, 18.02 mmol) in anhydrous DCM (40 mL) was added TEA (5.02 mL, 36.0 mmol) dropwise over 15 minutes at -70 °C. To this mixture was added a solution of phenyl phosphorodichloride (3.80 g, 18.02 mmol) in anhydrous DCM (50 mL) over 1 hour. The reaction mixture was stirred at this temperature for an additional 30 minutes, then warmed to 0 °C over 2 hours and stirred for 1 hour. To this mixture was added a solution of 2,3,4,5,6-pentafluorophenol (3.32 g, 18.02 mmol) and triethylamine (2.51 mL, 18.02 mmol) in DCM (30 mL) over 20 minutes and stirred at 0 °C for 16 hours. LCMS indicated the reaction was complete. The white solid (triethylamine hydrochloride) was filtered off and washed with DCM (50 mL). The filtrate was concentrated under reduced pressure, the residue was triturated with tert-butyl methyl ether (150 mL), and triethylamine hydrochloride was removed by filtration. The cake was washed with tert-butyl methyl ether (2 x 50 mL), and the combined filtrates were concentrated under reduced pressure to give a solid crude product (22 g) containing an even mixture of diastereomers. The crude product was triturated with 10% EtOAc / hexane (100 mL), and the solid was collected by filtration to give octyl ((perfluorophenoxy)(phenoxy)phosphoryl)-L-phenylalaninate (2,300 mg, 3.84 mmol, 21.28% yield) as a white solid. 31 >98% de as determined by P-NMR). 1H NMR (400MHz, CDCl3) δ 7.35 (t, J = 7.9 Hz, 2H), 7.22 (dt, J = 13.3, 5.7 Hz, 6H), 7.08 - 6.99 (m, 2H), 4.44 (ddd, J = 15.7, 10.0, 5.8 Hz, 1H), 4.16 - 3.95 (m, 2H), 3.77 (t, J = 11.1 Hz, 1H), 3.15 (dd, J = 13.7, 5.3 Hz, 1H), 3.05 (dd, J = 13.7, 6.3 Hz, 1H), 1.60 - 1.49 (m, 2H), 1.37 - 1.12 (m, 10H), 0.88 (t, J = 6.9 Hz, 3H). 31 P NMR (162MHz, CDCl3) δ -1.58.

[0243] Step 4: Octyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a stirred solution of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (0.18 g, 0.614 mmol) in THF (36 mL) and pyridine (9.00 mL) was added tert-butylmagnesium chloride (1.289 mL, 1.289 mmol) dropwise at −15° C. After 30 min, a solution of octyl ((S)-(perfluorophenoxy)(phenoxy)phosphoryl)-L-phenylalaninate (0.552 g, 0.921 mmol) in THF (5 mL) was added dropwise to the above solution at −15° C. and stirred at −15° C. for 16 h. LCMS indicated the reaction was complete. The reaction mixture was quenched with saturated NH4Cl and extracted with EtOAc (20 mL × 3). The combined organic phases were washed with brine (20 mL), dried over NaSO, filtered, and concentrated in vacuo. The crude product was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 40 g, 0-100% CHCN / 10 mM NHHCO in water) to give octyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (170 mg, 0.233 mmol, 38.0% yield) as a white solid. LCMS (M+H) = 709.0; retention time (10 mM NHHCO) = 1.974 min.

[0244] Step 5: (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-(octyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl decanoate To a stirred solution of decanoic acid (45.5 mg, 0.264 mmol) in DCM (4 mL), DMAP (147 mg, 1.199 mmol) and EDC (230 mg, 1.199 mmol) were added, and the resulting mixture was stirred at room temperature for 0.5 h. Then, a solution of octyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (170 mg, 0.240 mmol) in DCM (2 mL) was added, and the resulting mixture was stirred at room temperature overnight. LCMS showed the reaction was complete. The mixture was diluted with water (10 mL) and extracted with DCM (20 mL × 3). The organic phases were combined, washed with brine (20 mL), dried over Na2SO4, and concentrated in vacuo. The residue was purified using reverse-phase chromatography (SepaFlash® C18 column, 0-100% CH3CN / 10 mM NH4HCO3 in water) to give (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-(octyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl decanoate (129 mg, 0.140 mmol, 58.4% yield) as a pale yellow solid. LCMS: Retention time (0.1% TFA) = 2.746 min. HPLC: Retention time (10 mM NH4HCO3) = 9.376 min. 1H NMR (400MHz, CDCl3) δ 8.03 (s, 1H), 7.25-7.18 (m, 5H), 7.15 (d, J = 8.5 Hz, 2H), 7.14-7.09 (m, 3H), 6.39 (t, J = 6.7 Hz, 1H), 5.99 (s, 2H), 5.56 (dd, J = 6.3, 4.5 Hz, 1H), 4.32 (ddd, J = 16.6, 10.3, 6.4 Hz, 1H), 4.25 (dd, J = 11.1, 6.3 Hz, 1H), 4.13 (dd, J = 11.1, 5.7 Hz, 1H), 4.02 (t, J = 6.8 Hz, 2H), 3.63 (t, J = 10.7 Hz, 1H), 3.00 (d, J = 6.6 Hz, 2H), 2.62 (s, 1H), 2.62-2.57 (m, 2H), 2.40 (t, J = 7.6 Hz, 2H), 1.70-1.62 (m, 2H), 1.55-1.48 (m, 2H), 1.32-1.26 (m, 12H), 1.24 (s, 10H), 0.88 (t, J = 6.7 Hz, 6H).

[0245] Example 15

[0246]

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[0247] Process 1: Tert-Turbo-L-Fluor To a stirred mixture of (tert-butoxycarbonyl)-L-phenylalanine (15 g, 56.5 mmol), HOBt (11.46 g, 85 mmol), and EDC (13.01 g, 67.8 mmol) in DCM (150 mL) was added TEA (7.88 mL, 56.5 mmol) at 0 °C. After stirring for 30 min, dodecan-1-ol (10.54 g, 56.5 mmol) was added. The resulting reaction mixture was stirred at 25 °C for 16 h. LCMS showed the reaction was complete. The reaction mixture was diluted with water (80 mL) and extracted with DCM (100 mL × 2). The combined organic layers were washed with brine, dried over Na2SO4, and concentrated. The residue was purified using silica gel chromatography (petroleum ether: EtOAc = 10:1) to give dodecyl (tert-butoxycarbonyl)-L-phenylalaninate (14.2 g, 32.2 mmol, 57.0% yield) as a yellow oil. LCMS (M + Na) = 456.4; retention time (0.05% TFA) = 3.20 min.

[0248] Step 2: Dodecyl L-phenylalaninate To a stirred solution of dodecyl (tert-butoxycarbonyl)-L-phenylalaninate (14.6 g, 33.7 mmol) in DCM (150 mL) was added TFA (31.1 mL, 404 mmol) at 0 °C. The resulting reaction mixture was stirred at 25 °C for 4 h. LCMS indicated the reaction was complete. The reaction mixture was concentrated, and the residue was diluted with DCM (100 mL) and neutralized with NaHCO (50 mL). The organic layer was separated, and the aqueous layer was extracted with DCM (2 × 100 mL). The combined organic layers were washed with brine (80 mL), dried over NaSO, filtered, and concentrated. The residue was purified using silica gel chromatography (petroleum ether: EtOAc = 2:1) to afford dodecyl L-phenylalaninate (9.4 g, 27.6 mmol, 82% yield) as a colorless oil. LCMS (M+H)=334.3; Retention time (0.05%TFA)=1.88 min.

[0249] Step 3: Dodecyl ((S)-(perfluorophenoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a stirred solution of dodecyl L-phenylalaninate (4 g, 11.99 mmol) in anhydrous DCM (25 mL) was added triethylamine (1.834 mmol, 13.19 mmol) dropwise at -70 °C. To this mixture was added a solution of phenyl phosphorodichloridate (2.505 g, 11.87 mmol) in anhydrous DCM (8 mL) dropwise. The reaction mixture was stirred at this temperature for an additional 30 minutes, then warmed to 0 °C over 2 hours and stirred for 1 hour. To this mixture was added a solution of 2,3,4,5,6-pentafluorophenol (2.185 g, 11.87 mmol) and triethylamine (1.834 mL, 13.19 mmol) in DCM (30 mL) dropwise and stirred at 0 °C for 2 hours. LCMS indicated the reaction was complete. The white solid (triethylamine hydrochloride) was filtered off and washed with DCM (10 mL). The filtrate was concentrated under reduced pressure, the residue was triturated with tert-butyl methyl ether (20 mL), and triethylamine hydrochloride was removed by filtration. The cake was washed with tert-butyl methyl ether (2 × 10 mL), and the filtrate was concentrated under reduced pressure. The mixture was triturated with 10% EtOAc / hexane (30 mL), and the solid was collected by filtration to give dodecyl ((S)-(perfluorophenoxy)(phenoxy)phosphoryl)-L-phenylalaninate (1.8 g, 2.57 mmol, 21.39% yield) as a white solid. 31 >98% de by P-NMR. LCMS (M+H) = 656.2; retention time (0.05% TFA) = 3.31 min. 31 P NMR (162MHz, DMSO-d6) δ 0.21.

[0250] Step 3: Dodecyl ((R)-(perfluorophenoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a stirred solution of dodecyl L-phenylalaninate (5.9 g, 17.69 mmol) in anhydrous DCM (40 mL) was added TEA (2.71 mL, 19.46 mmol) dropwise over 15 min at -70 °C. To this mixture was added a solution of phenyl phosphorodichloride (3.73 g, 17.69 mmol) in anhydrous DCM (50 mL) over 1 h. The reaction mixture was stirred at this temperature for an additional 30 min, then warmed to 0 °C over 2 h and stirred for 1 h. To this mixture was added a solution of 2,3,4,5,6-pentafluorophenol (3.26 g, 17.69 mmol) and triethylamine (2.71 mL, 19.46 mmol) in DCM (30 mL) over 20 min and stirred at 0 °C for 16 h. TLC indicated the reaction was complete. The white solid (triethylamine hydrochloride) was filtered off and washed with DCM (50 mL). The filtrate was concentrated under reduced pressure, the residue was triturated with tert-butyl methyl ether (150 mL), and triethylamine hydrochloride was removed by filtration. The cake was washed with tert-butyl methyl ether (2 x 50 mL), and the combined filtrates were concentrated under reduced pressure to give a solid containing an equal mixture of diastereomers. The crude product was triturated with 20% EtOAc / hexane (100 mL), and the solid was collected by filtration to give dodecyl ((R)-(perfluorophenoxy)(phenoxy)phosphoryl)-L-phenylalaninate (2.3 g, 3.51 mmol, 19.83% yield, mp 180-185°C). 31 P-NMR analysis revealed that >98% de 1 H NMR (400MHz, CDCl3) δ 7.40 - 7.30 (m, 2H), 7.25 - 7.15 (m, 6H), 7.09 - 7.03 (m, 2H), 4.52 - 4.36 (m, 1H), 4.16 - 3.97 (m, 2H), 3.88 (dd, J = 19.3, 10.1 Hz, 1H), 3.14 (dd, J = 13.7, 5.4 Hz, 1H), 3.10 - 2.99 (m, 1H), 1.61 - 1.46 (m, 2H), 1.38 - 1.16 (m, 18H), 0.88 (t, J = 6.7 Hz, 3H). 31P NMR (162MHz, CDCl3) δ -1.56.

[0251] Step 4: Dodecyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a stirred solution of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (300 mg, 1.023 mmol) in anhydrous THF (60 mL) and pyridine (15.0 mL) was added tert-butylmagnesium chloride (2.046 mL, 2.046 mmol) dropwise at −15° C. To this mixture was added a solution of dodecyl ((S)-(perfluorophenoxy)(phenoxy)phosphoryl)-L-phenylalaninate (939 mg, 1.432 mmol) in anhydrous THF (20 mL) dropwise over 1 h. The reaction mixture was stirred at this temperature overnight. LCMS indicated the reaction was complete. The reaction mixture was quenched with 2N NH4Cl (10 mL) and water (50 mL) and extracted with EtOAc (2 x 150 mL). The combined organic layers were then washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified using reverse-phase column chromatography (SepaFlash® C18 column, BOSTON, 80 g, 0-100% CH3OH / 10 mM NH4HCO3 aqueous solution) to afford dodecyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (190 mg, 0.248 mmol, 24.28% yield) as a white solid. LCMS (M+H)=764.9; retention time (10mM NH4HCO3)=2.44 min.

[0252] Step 4: Dodecyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a stirred solution of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (0.36 g, 1.228 mmol) in THF (100 mL) and pyridine (2.00 mL) was added tert-butylmagnesium chloride (2.58 mL, 2.58 mmol) dropwise at 0°C, and the reaction was stirred at 25°C for 30 min. A solution of dodecyl ((S)-(perfluorophenoxy)(phenoxy)phosphoryl)-L-phenylalaninate (0.805 g, 1.228 mmol) in THF (5 mL) was added dropwise to the above solution at -15°C, and the mixture was stirred at -15°C for 4 h. TLC showed the presence of a new compound. The reaction was quenched with 2M HCl (10 mL) and extracted with EtOAc (50 mL × 3). The combined organic phase was washed with NaHCO3 solution (50 mL), washed with brine (20 mL), dried over Na2SO4, filtered, and concentrated in vacuo. The crude product was purified using silica gel chromatography (DCM:MeOH = 20:1) to afford dodecyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (270 mg, 0.353 mmol, 28.8% yield) as a white solid. LCMS (M+H) + =765.7; Retention time (0.1%TFA)=2.41 minutes.

[0253] Step 5: (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((S)-(((S)-1-(dodecyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl decanoate To a cold (ice-water bath) stirred solution of decanoic acid (37.2 mg, 0.216 mmol) in DCM (15 mL) was added 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (188 mg, 0.981 mmol) and N,N-dimethylpyridin-4-amine (120 mg, 0.981 mmol). After 0.5 h, dodecyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (150 mg, 0.196 mmol) was added, and the resulting mixture was stirred at 25 °C for 16 h. TLC indicated the reaction was complete. The reaction mixture was partitioned between DCM (100 mL) and water (30 mL). The organic layer was then washed with brine (15 mL), dried over NaSO, filtered, and concentrated. The residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 40 g, 0-100% CHOH / 10 mM NHHCO aqueous solution) to afford (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((S)-(((S)-1-(dodecyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl decanoate (129 mg, 0.138 mmol, 70.5% yield) as a colorless oil. LCMS (M+H)=919.3; Retention time (0.1%TFA)=3.52 minutes. Purity: 100% (254nm). HPLC retention time (10mM NH4HCO3) = 11.12 minutes. 1H NMR (400MHz, CDCl3) δ 7.99 (s, 1H), 7.25-7.09 (m, 10H), 6.40 (t, J = 6.8 Hz, 1H), 5.99 (brs, 2H), 5.56 (dd, J = 6.4, 4.4 Hz, 1H), 4.36-4.29 (m, 1H), 4.25 (dd, J = 11.2, 6.4 Hz, 1H), 4.13 (dd, J = 10.8, 5.6 Hz, 1H), 4.02 (t, J = 6.8 Hz, 2H), 3.64 (t, J = 10.8 Hz, 1H), 3.00 (d, J = 6.4 Hz, 2H), 2.67-2.56 (m, 3H), 2.40 (t, J = 7.6 Hz, 2H), 1.67 (quint, J = 7.2 Hz, 2H), 1.52 (quint, J = 7.2 Hz, 2H), 1.30-1.20 (m, 30H), 0.91-0.86 (m, 6H).

[0254] Example 16

[0255]

change

[0256] Project 1: Tert-ブトキシカルボニル-L-フェニルアラニネート To a stirred mixture of (tert-butoxycarbonyl)-L-phenylalanine (15 g, 56.5 mmol), HOBt (4.33 g, 28.3 mmol), and EDC (16.26 g, 85 mmol) in DCM (120 mL) was added DIPEA (29.6 mL, 170 mmol) at 0 °C. After 30 min, a solution of tetradecan-1-ol (12.12 g, 56.5 mmol) in DCM (20 mL) was added and stirred at 25 °C for 16 h. LCMS indicated the presence of a new compound. The reaction mixture was diluted with water (80 mL), the organic layer was separated, and the aqueous phase was extracted with DCM (80 mL × 2). The combined organic phase was washed with brine, dried over Na2SO4, and concentrated. The residue was purified using silica gel chromatography (Santai, 120 g, petroleum ether: EtOAc = 30:1) to give tetradecyl (tert-butoxycarbonyl)-L-phenylalaninate (20 g, 39.4 mmol, 69.8% yield) as a yellow oil. LCMS (M + Na) = 484.2; retention time (10% TFA) = 3.532 min.

[0257] Step 2: Tetradecyl L-phenylalaninate To a stirred mixture of tetradecyl (tert-butoxycarbonyl)-L-phenylalaninate (20 g, 43.3 mmol) in DCM (160 mL) was added TFA (28.6 mL, 372 mmol) at 0 °C and stirred at 25 °C for 4 h. LCMS indicated the presence of a new compound. The reaction was concentrated, the pH of the residue was adjusted to 8–9 with 1N NaOH, and the mixture was extracted with DCM (100 mL × 3). The combined organic phases were washed with brine, dried over Na2SO4, and concentrated. The residue was purified using silica gel chromatography (Santai, 80 g, DCM:MeOH = 10:1) to give tetradecyl L-phenylalaninate (12 g, 33.2 mmol, 77% yield) as a yellow oil. LCMS (M+H) = 362.4; retention time (0.1% TFA) = 2.506 min.

[0258] Step 3: Tetradecyl ((perfluorophenoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a stirred solution of tetradecyl L-phenylalaninate (6.0 g, 16.59 mmol) in anhydrous DCM (100 mL) was added triethylamine (2.54 mL, 18.25 mmol) dropwise over 15 minutes at -70 °C. To this mixture was added a solution of phenyl phosphorodichloridate (3.47 g, 16.43 mmol) in anhydrous DCM (35 mL) over 1 hour. The reaction mixture was stirred at this temperature for an additional 30 minutes, then warmed to 0 °C over 2 hours and stirred for 1 hour. To this mixture was added a solution of 2,3,4,5,6-pentafluorophenol (3.02 g, 16.43 mmol) and triethylamine (2.54 mL, 18.25 mmol) in DCM (30 mL) over 20 minutes and stirred at 0 °C for 4 hours. LCMS indicated the reaction was complete. The white solid (triethylamine hydrochloride) was filtered off and washed with DCM (50 mL). The filtrate was concentrated under reduced pressure, and the residue was triturated with tert-butyl methyl ether (250 mL), and then the triethylamine hydrochloride was removed by filtration. The cake was washed with tert-butyl methyl ether (2 x 50 mL), and the combined filtrates were concentrated under reduced pressure to give 26 g of crude solid containing an equal mixture of diastereomers. The crude product was triturated with 20% EtOAc / hexane (200 mL), and the solid was collected by filtration to give tetradecyl ((S)-(perfluorophenoxy)(phenoxy)phosphoryl)-L-phenylalaninate (5.0 g, 7.31 mmol, 44.1% yield) as a white solid. 31 >98% de:N76107-92-A1) as determined by P-NMR. 1 H NMR (400MHz, CDCl3) δ 7.35 (t, J = 7.9 Hz, 2H), 7.26 - 7.15 (m, 6H), 7.12 - 7.00 (m, 2H), 4.50 - 4.38 (m, 1H), 4.15 - 3.95 (m, 2H), 3.85 - 3.70 (m, 1H), 3.10 (ddd, J = 37.7, 13.4, 5.5 Hz, 2H), 1.54 (s, 2H), 1.36 - 1.14 (m, 22H), 0.88 (t, J = 6.8 Hz, 3H). 31P NMR (162 MHz, CDCl3) δ -1.57.

[0259] Step 4: Tetradecyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a stirred solution of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (0.6 g, 2.046 mmol) in THF (120 mL) and pyridine (30.0 mL) was added tert-butylmagnesium chloride (4.30 mL, 4.30 mmol) dropwise at −15° C. The reaction was then stirred at 25° C. for 30 minutes. A solution of tetradecyl ((S)-(perfluorophenoxy)(phenoxy))-L-phosphorylphenylalaninate (2.098 g, 3.07 mmol) in THF (12.5 mL) was added dropwise to the above solution at −15° C. and stirred at −15° C. for 4 hours. LCMS indicated the reaction was complete. The reaction was quenched with saturated NH4Cl (20 mL) and concentrated to remove THF. The resulting aqueous mixture was extracted with EtOAc (20 mL × 3), and the combined organic phases were washed with NH4Cl solution (20 mL), brine (20 mL), dried over Na2SO4, filtered, and concentrated in vacuo. The crude product was purified using reverse-phase chromatography (SepaFlash® C18 column, 80 g, 0–100% CH3CN / 10 mM NH4HCO3 aqueous solution) to afford tetradecyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (760 mg, 0.857 mmol, 41.9% yield) as a white solid. LCMS (M+H)=793.0; Retention time (10mM NH4HCO3)=2.767 minutes.

[0260] Step 5: (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-oxo-3-phenyl-1-(tetradecyloxy)propan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl decanoate To a stirred solution of decanoic acid (47.8 mg, 0.277 mmol) in DCM (4 mL), DMAP (154 mg, 1.261 mmol) and EDC (242 mg, 1.261 mmol) were added and stirred at room temperature for 0.5 h. Tetradecyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (200 mg, 0.252 mmol) was then added and stirred at room temperature for 16 h. TLC (DCM:MeOH = 17:1, Rf = 0.45) indicated the presence of a new compound. The mixture was quenched with water (10 mL) and extracted with DCM (20 mL × 3). The combined organic phase was washed with brine, dried over Na2SO4, and concentrated in vacuo. The residue was purified using preparative TLC (DCM:MeOH = 17:1, Rf = 0.45) to give (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-oxo-3-phenyl-1-(tetradecyloxy)propan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl decanoate (137 mg, 0.139 mmol, 55.2% yield) as a pale yellow solid. LCMS (M+H) = 947.7; retention time (0.1% TFA) = 4.127 min. HPLC; retention time (10 mM NH4HCO3) = 14.764 min. 1H NMR (400MHz, CDCl3) δ 8.01 (s, 1H), 7.25-7.21 (d, J = 7.6 Hz, 4H), 7.20 (dd, J = 6.4, 2.1 Hz, 1H), 7.15 (d, J = 8.6 Hz, 2H), 7.13-7.08 (m, 3H), 6.39 (t, J = 6.7 Hz, 1H), 5.99 (s, 2H), 5.57 (dd, J = 6.6, 4.3 Hz, 1H), 4.32 (ddd, J = 9.6, 8.1, 4.9 Hz, 1H), 4.25 (dd, J = 11.1, 6.3 Hz, 1H), 4.13 (dd, J = 11.1, 5.7 Hz, 1H), 4.02 (t, J = 6.8 Hz, 2H), 3.63 (t, J = 10.7 Hz, 1H), 3.00 (d, J = 6.4 Hz, 2H), 2.66-2.54 (m, 3H), 2.40 (t, J = 7.6 Hz, 2H), 1.71-1.62 (m, 2H), 1.55-1.48 (m, 2H), 1.27 (d, J = 2.5 Hz, 9H), 1.25 (d, J = 4.9 Hz, 24H), 0.90-0.86 (m, 6H).

[0261] Example 17

[0262] [ka]

[0263] (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-(octyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl icosanoate To a solution of icosanoic acid (97 mg, 0.310 mmol) in DCM (4 mL), DMAP (172 mg, 1.411 mmol) and EDC (270 mg, 1.411 mmol) were added and stirred at room temperature for 0.5 h. Then, a solution of octyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (N77883-37, 200 mg, 0.282 mmol) in DCM (3 mL) was added and stirred at room temperature for 16 h. TLC (DCM:MeOH = 17:1, Rf = 0.45) indicated the presence of a new compound. The reaction mixture was quenched with water (10 mL) and extracted with DCM (20 mL × 3). The combined organic phase was washed with brine, dried over NaSO, and concentrated in vacuo. The residue was purified using preparative TLC (DCM:CHOH=17:1, Rf=0.45) to give (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-(octyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl icosanoate (170 mg, 0.162 mmol, 57.4% yield) as a white solid. HPLC; Retention time (10 mM NHHCO): 15.683 min. 1H NMR (400MHz, CDCl3) δ 8.00 (s, 1H), 7.22 (d, J = 7.9 Hz, 4H), 7.20-7.18 (m, 1H), 7.15 (d, J = 8.6 Hz, 2H), 7.13-7.08 (m, 3H), 6.39 (t, J = 6.7 Hz, 1H), 6.08 (s, 2H), 5.57 (dd, J = 6.6, 4.2 Hz, 1H), 4.36-4.28 (m, 1H), 4.25 (dd, J = 11.1, 6.3 Hz, 1H), 4.13 (dd, J = 11.1, 5.7 Hz, 1H), 4.02 (t, J = 6.8 Hz, 2H), 3.68 (t, J = 10.7 Hz, 1H), 3.00 (d, J = 6.4 Hz, 2H), 2.67-2.53 (m, 3H), 2.39 (t, J = 7.6 Hz, 2H), 1.71-1.61 (m, 2H), 1.56-1.47 (m, 2H), 1.25 (d, J = 4.2 Hz, 42H), 0.88 (t, J = 6.7 Hz, 6H).

[0264] Example 18

[0265]

change

[0266] Process 1: Tactical ((S)-(パーフルオロフェノキシ) (フェノキシ)ホスホリル)-L-フェニルアラニネート To a solution of decyl L-phenylalaninate (3 g, 9.82 mmol) in DCM (20 mL) was added triethylamine (1.502 mL, 10.80 mmol) dropwise at -70 °C. To this mixture was added a solution of phenyl phosphorodichloridate (2.051 g, 9.72 mmol) in anhydrous DCM (8 mL) dropwise. The reaction mixture was stirred at this temperature for an additional 30 minutes, then warmed to 0 °C over 2 hours and stirred for 1 hour. To this mixture was added a solution of 2,3,4,5,6-pentafluorophenol (1.790 g, 9.72 mmol) and triethylamine (1.502 mL, 10.80 mmol) in DCM (30 mL) dropwise and stirred at 0 °C for 2 hours. LCMS indicated the reaction was complete. The white solid (triethylamine hydrochloride) was filtered off and washed with DCM (10 mL). The filtrate was concentrated under reduced pressure, the residue was triturated with tert-butyl methyl ether (20 mL), and triethylamine hydrochloride was removed by filtration. The cake was washed with tert-butyl methyl ether (2 × 10 mL), and the filtrate was concentrated under reduced pressure. The residue was triturated with 10% EtOAc / hexane (30 mL), and the solid was collected by filtration to give decyl ((S)-(perfluorophenoxy)(phenoxy)phosphoryl)-L-phenylalaninate (1.5 g, 2.390 mmol, 24.34% yield) as a white solid. 31 P-NMR: >98% de (LCMS (M+H) = 628.0; retention time (0.1% TFA) = 2.78 min). 31 P NMR (DMSO-d6, 162MHz) δ 0.22.

[0267] Step 2: Decyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a solution of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (150 mg, 0.511 mmol) in anhydrous THF (30 mL) and pyridine (15 mL) was added tert-butylmagnesium chloride (1.023 mL, 1.023 mmol) dropwise at −15° C. The reaction mixture was stirred at the same temperature for 1 hour. Then, a solution of decyl ((S)-(perfluorophenoxy)(phenoxy)phosphoryl)-L-phenylalaninate (642 mg, 1.023 mmol) in anhydrous THF (15 mL) was added dropwise at −15° C., followed by stirring at −15° C. for 16 hours. LCMS indicated the reaction was complete. The reaction mixture was quenched with 2N NH4Cl (20 mL) and diluted with EtOAc (200 mL). The organic layer was washed with water (40 mL), brine (40 mL), dried over NaSO, filtered, and concentrated. The residue was purified by reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 40 g, 0-100% CHOH / 10 mM NH4HCO3 aqueous solution) to give decyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (150 mg, 0.196 mmol, 38.4% yield) as a white solid. LCMS (M+H)=737.0; Retention time (10mM NH4HCO3)=2.20 minutes.

[0268] Step 3: (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((S)-(((S)-1-(decyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl icosanoate To a cold (ice-water bath) stirred solution of icosanoic acid (63.6 mg, 0.204 mmol) in DCM (10 mL), 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (195 mg, 1.018 mmol) and N,N-dimethylpyridin-4-amine (124 mg, 1.018 mmol) were added and stirred for 0.5 h. Decyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (150 mg, 0.204 mmol) was then added and stirred at 25° C. for 16 h. TLC indicated the reaction was complete. The reaction mixture was partitioned between DCM (100 mL) and water (30 mL). The organic layer was then washed with brine (20 mL), dried over NaSO, filtered, and concentrated. The residue was purified using preparative TLC (DCM / CHOH = 20 / 1) to afford (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((S)-(((S)-1-(decyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl icosanoate (133 mg, 0.127 mmol, 62.5% yield) as a colorless oil. HPLC retention time (10 mM NH4HCO3) = 19.81 min. 1H NMR (400MHz, CDCl3) δ 8.02 (s, 1H), 7.26-7.14 (m, 7H), 7.13-7.09 (m, 3H), 6.40 (t, J = 6.8 Hz, 1H), 6.11 (brs, 2H), 5.56 (dd, J = 6.4, 4.4 Hz, 1H), 4.36-4.29 (m, 1H), 4.25 (dd, J = 11.2, 6.0 Hz, 1H), 4.13 (dd, J = 10.8, 5.6 Hz, 1H), 4.02 (t, J = 6.8 Hz, 2H), 3.69 (t, J = 10.8 Hz, 1H), 3.00 (d, J = 6.4 Hz, 2H), 2.67-2.55 (m, 3H), 2.40 (t, J = 7.6 Hz, 2H), 1.67 (quint, J = 7.2 Hz, 2H), 1.52 (quint, J = 7.2 Hz, 2H), 1.31-1.20 (m, 46H), 0.88 (t, J = 6.8 Hz, 6H).

[0269] Example 19

[0270] [ka]

[0271] (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((S)-(((S)-1-(dodecyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl icosanoate To a cold (ice-water bath) stirred solution of icosanoic acid (57.2 mg, 0.183 mmol) in DCM (15 mL), 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (175 mg, 0.915 mmol) and N,N-dimethylpyridin-4-amine (112 mg, 0.915 mmol) were added, and the resulting mixture was stirred for 0.5 h. Dodecyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (N76670-81, 140 mg, 0.183 mmol) was then added and stirred at room temperature for 4 h. TLC indicated the reaction was complete. The reaction mixture was partitioned between DCM (100 mL) and water (30 mL). The organic layer was washed with brine, dried over NaSO, filtered, and concentrated. The residue was purified using preparative TLC (DCM / CHOH=20 / 1) to afford (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((S)-(((S)-1-(dodecyloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl icosanoate (103 mg, 0.092 mmol, 50.4% yield) as a colorless oil. HPLC retention time (10 mM NHHCO) = 26.12 min. 1H NMR (400MHz, CDCl3) δ 8.02 (s, 1H), 7.29-7.27 (m, 1H), 7.25-7.19 (m, 4H), 7.18-7.14 (m, 2H), 7.13-7.09 (m, 3H), 6.40 (t, J = 6.8 Hz, 1H), 5.99 (brs, 2H), 5.56 (dd, J = 6.0, 4.4 Hz, 1H), 4.36-4.29 (m, 1H), 4.25 (dd, J = 11.2, 6.4 Hz, 1H), 4.13 (dd, J = 10.8, 5.6 Hz, 1H), 4.02 (t, J = 6.8 Hz, 2H), 3.63 (t, J = 10.4 Hz, 1H), 3.00 (d, J = 6.8 Hz, 2H), 2.65-2.57 (m, 3H), 2.40 (t, J = 7.6 Hz, 2H), 1.67 (quint, J = 7.2 Hz, 2H), 1.52 (quint, J = 6.0 Hz, 2H), 1.30-1.23 (m, 50H), 0.88 (t, J = 6.8 Hz, 6H).

[0272] Example 20

[0273] [ka]

[0274] (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-oxo-3-phenyl-1-(tetradecyloxy)propan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl icosanoate To a stirred solution of icosanoic acid (87 mg, 0.277 mmol) in DCM (4 mL), DMAP (154 mg, 1.261 mmol) and EDC (242 mg, 1.261 mmol) were added at 0 °C, and the resulting mixture was stirred for 0.5 h. Then, a solution of tetradecyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (N77883-38, 200 mg, 0.252 mmol) in DCM (3 mL) was added at 0 °C. The resulting mixture was stirred overnight at room temperature. TLC (DCM:CHOH = 17:1, Rf = 0.45) indicated the presence of a new compound. The mixture was quenched with water (10 mL) and extracted with DCM (20 mL × 3). The combined organic phase was washed with brine, dried over Na2SO4, and concentrated in vacuo. The residue was purified using preparative TLC (DCM:CH3OH = 17:1, Rf = 0.45) to give (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-oxo-3-phenyl-1-(tetradecyloxy)propan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl icosanoate (172 mg, 0.149 mmol, 59.0% yield) as a white solid. HPLC: Retention time (10 mM NH4HCO3): 37.405 min. 1H NMR (400MHz, CDCl3) δ 8.03 (s, 1H), 7.25-7.21 (m, 4H), 7.19 (d, J = 6.9 Hz, 1H), 7.15 (d, J = 8.5 Hz, 2H), 7.08-7.03 (m, 3H), 6.39 (t, J = 6.6 Hz, 1H), 6.04 (s, 2H), 5.56 (t, J = 5.4 Hz, 1H), 4.36-4.28 (m, 1H), 4.25 (dd, J = 11.1, 6.3 Hz, 1H), 4.13 (dd, J = 11.1, 5.7 Hz, 1H), 4.02 (t, J = 6.7 Hz, 2H), 3.64 (t, J = 10.7 Hz, 1H), 3.00 (d, J = 5.9 Hz, 2H), 2.64-2.54 (m, 3H), 2.39 (t, J = 7.6 Hz, 2H), 1.71-1.62 (m, 2H), 1.55-1.48 (m, 2H), 1.28 (d, J = 17.2 Hz, 54H), 0.88 (t, J = 6.8 Hz, 6H).

[0275] Example 21

[0276]

change

[0277] Project 1: Tert-ブトキシカルボニル-L-フェニルアラニネート To a stirred mixture of (tert-butoxycarbonyl)-L-phenylalanine (40 g, 151 mmol) in DCM (300 mL) was added DMAP (1.842 g, 15.08 mmol) and EDC (37.6 g, 196 mmol) at 0 °C. After 0.5 h, a solution of heptan-4-ol (22.80 g, 196 mmol) in DCM (50 mL) was added at 0 °C. The mixture was stirred at room temperature for 2 h. LCMS indicated the presence of a new compound. The reaction mixture was diluted with water (90 mL), the organic layer was separated, and the aqueous phase was extracted with DCM (100 mL × 3). The combined organic phase was washed with brine, dried over Na2SO4, and concentrated. The residue was purified using silica gel chromatography (Santai, 220 g, petroleum ether: EtOAc = 30:1) to give heptan-4-yl (tert-butoxycarbonyl)-L-phenylalaninate (45 g, 124 mmol, 82% yield) as a pale yellow oil. LCMS (M + Na) = 386.1; retention time (10% TFA) = 2.36 min.

[0278] Step 2: Heptan-4-yl L-phenylalaninate To a stirred mixture of heptan-4-yl (tert-butoxycarbonyl)-L-phenylalaninate (45 g, 124 mmol) in DCM (300 mL) was added TFA (60 mL, 779 mmol) at 0 °C, and the resulting mixture was stirred at 25 °C for 5 h. LCMS showed the reaction was complete. The reaction was concentrated, and the pH of the residue was adjusted to 8–9 with NaOH (1 N) and extracted with DCM (100 mL × 3). The combined organic phase was washed with brine, dried over NaSO, and concentrated. The residue was purified using silica gel chromatography (Santai, 220 g, DCM:MeOH = 10:1) to give heptan-4-yl L-phenylalaninate (30 g, 114 mmol, 92% yield) as a pale yellow oil. LCMS(M+H)=264.0; Time(0.1%TFA)=1.516 min.

[0279] Step 3: Heptan-4-yl ((perfluorophenoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a solution of heptan-4-yl L-phenylalaninate (18.5 g, 70.2 mmol) in anhydrous DCM (35 mL) was added triethylamine (10.25 mL, 73.8 mmol) dropwise over 15 minutes at -70°C. To this mixture was added a solution of phenyl phosphorodichloridate (14.67 g, 69.5 mmol) in anhydrous DCM (35 mL) over 1 hour. The reaction mixture was stirred at this temperature for an additional 30 minutes, then warmed to 0°C over 2 hours and stirred for 1 hour. To this mixture was added a solution of 2,3,4,5,6-pentafluorophenol (12.80 g, 69.5 mmol) and triethylamine (10.74 mL, 77 mmol) in DCM (30 mL) over 20 minutes and stirred at 0°C for 4 hours. LCMS indicated the reaction was complete. The white solid (triethylamine hydrochloride) was filtered off and washed with DCM (25 mL). The filtrate was concentrated under reduced pressure, the residue was triturated with tert-butyl methyl ether (250 mL), and the triethylamine hydrochloride was removed by filtration. The cake was washed with tert-butyl methyl ether (2 x 50 mL), and the filtrate was concentrated under reduced pressure to give 44 g of crude product. The crude product was triturated with 5% EtOAc / hexane (200 mL), and the solid was collected by filtration to give heptan-4-yl ((perfluorophenoxy)(phenoxy)phosphoryl)-L-phenylalaninate (5 g, 7.19 mmol, 10.24% yield) as a white solid. 31 P-NMR determined to be >98% de), LCMS (M+H) = 586.2; retention time (0.1% TFA) = 2.137 min. 1 H NMR (400MHz, CDCl3) δ 7.33 (t, J = 7.9 Hz, 2H), 7.23 (t, J = 5.5 Hz, 3H), 7.21-7.15 (m, 3H), 7.12-7.08 (m, 2H), 4.91-4.83 (m, 1H), 4.42 (tt, J = 9.8, 6.1 Hz, 1H), 3.59 (s, 1H), 3.10 (d, J = 6.0 Hz, 2H), 1.50-1.39 (m, 4H), 1.26-1.16 (m, 4H), 0.84 (td, J = 7.3, 5.3Hz, 6H). 31P NMR (162MHz, CDCl3) δ -1.62.

[0280] Step 2: Heptan-4-yl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a stirred solution of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (800 mg, 2.73 mmol) in THF (160 mL) and pyridine (20 mL) was added tert-butylmagnesium chloride (5.46 mL, 5.46 mmol) dropwise at −15° C. The reaction mixture was stirred at the same temperature for 1 hour. Then, to this reaction mixture was added dropwise a solution of heptan-4-yl ((S)-(perfluorophenoxy)(phenoxy)phosphoryl)-L-phenylalaninate (2396 mg, 4.09 mmol) in anhydrous THF (10 mL), and the mixture was stirred at −15° C. for 4 hours. LCMS showed the reaction was complete. The reaction mixture was quenched with 2N NH4Cl (20 mL) and partitioned between EtOAc (200 mL) and water (50 mL). The aqueous layer was extracted with EtOAc (2 × 150 mL), and the combined organic layers were washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 120 g, 0–100% CH3CN / 10 mM NH4HCO3 in water) to afford heptan-4-yl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (700 mg, 0.984 mmol, 36.1% yield) as a white solid. LCMS (M+H)=695.3; Retention time (0.05%TFA)=1.78 min.

[0281] Step 5: (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-(heptan-4-yloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl 2-propylpentanoate To a stirred solution of 2-propylpentanoic acid (37.7 mg, 0.261 mmol) in DCM (4 mL) was added DMAP (145 mg, 1.188 mmol) and EDC (228 mg, 1.188 mmol) at 0° C. After 0.5 h, heptan-4-yl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (165 mg, 0.238 mmol) was added, and the resulting mixture was stirred at 15° C. for 16 h. LCMS indicated the reaction was complete. The mixture was diluted with water (20 mL) and extracted with DCM (20 mL × 2). The combined organic phase was washed with brine (20 mL), dried over NaSO, and concentrated in vacuo. The residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 12 g, 0-100% CHCN / 10 mM NH4HCO3 aqueous solution) to give (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-(heptan-4-yloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl 2-propylpentanoate (134 mg, 0.153 mmol, 64.5% yield) as a white solid. LCMS (M+H)=821.7; Retention time (0.1%TFA)=2.396 minutes. HPLC retention time (10mM NH4HCO3) = 7.883 minutes. 11H NMR (400 MHz, CDCl3) δ 7.99 (s, 1H), 7.25 - 7.21 (m, 4H), 7.20 - 7.17 (m, 1H), 7.17 - 7.13 (m, 4H), 7.10 (t, J = 7.3 Hz, 1H), 6.38 (t, J = 6.6 Hz, 1H), 5.98 (s, 2H), 5.53 (dd, J = 7.0, 4.7 Hz, 1H), 4.87 - 4.80 (m, 1H), 4.33 (tt, J = 10.1, 6.7 Hz, 1H), 4.20 (dd, J = 11.1, 6.2 Hz, 1H), 4.05 (dd, J = 11.1, 5.6 Hz, 1H), 3.65 (t, J = 10.5 Hz, 1H), 3.06 (dd, J = 13.8, 6.2 Hz, 1H), 2.94 (dd, J = 13.8, 6.9 Hz, 1H), 2.73 - 2.64 (m,1H), 2.61 (s, 1H), 2.57 (ddd, J = 13.9, 6.4, 4.7 Hz, 1H), 2.47 (tt, J = 8.6, 5.5 Hz, 1H), 1.70 - 1.60 (m, 2H), 1.52 - 1.45 (m, 2H), 1.45 - 1.38 (m, 4H), 1.34 (dd, J = 15.0, 7.4 Hz, 4H), 1.23 - 1.12 (m, 4H), 0.92 (td, J = 7.2, 1.6 Hz, 6H), 0.83 (td, J = 7.3, 1.5 Hz, 6H).

[0282] Example 22

[0283]

Chemical Structure

[0284] (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-(heptan-4-yloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl octanoate To a cold (ice-water bath) stirred solution of octanoic acid (37.4 mg, 0.259 mmol) in DCM (10 mL), 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (207 mg, 1.080 mmol) and N,N-dimethylpyridin-4-amine (132 mg, 1.080 mmol) were added and stirred for 0.5 h. Then, heptan-4-yl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (150 mg, 0.216 mmol) was added and stirred at 25 °C for 4 h. LCMS indicated the reaction was complete. The reaction mixture was partitioned between DCM (100 mL) and water (30 mL). The organic layer was washed with brine (30 mL), dried over NaSO, filtered, and concentrated. The residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 40 g, 0-100% CHCN / 10 mM NHHCO in water) to afford (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-(heptan-4-yloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl octanoate (107 mg, 0.124 mmol, 57.5% yield) as a white solid. LCMS (M+H)=821.3; Retention time (0.05%TFA)=2.14 minutes. HPLC retention time (10mM NH4HCO3) = 8.09 minutes. 1H NMR (400MHz, CDCl3) δ 8.00 (s, 1H), 7.26-7.21 (m, 4H), 7.19-7.09 (m, 6H), 6.39 (t, J = 6.8 Hz, 1H), 6.24 (brs, 2H), 5.55 (t, J = 5.2 Hz, 1H), 4.90-4.82 (m, 1H), 4.38-4.28 (m, 1H), 4.19 (dd, J = 10.8, 6.4 Hz, 1H), 4.06 (dd, J = 11.2, 5.6 Hz, 1H), 3.75 (t, J = 10.8 Hz, 1H), 3.07 (dd, J = 14.0, 6.4 Hz, 1H), 2.94 (dd, J = 14.0, 7.2 Hz, 1H), 2.62 (s, 1H), 2.60-2.55 (m, 2H), 2.40 (t, J = 7.6 Hz, 2H), 1.67 (quint, J = 7.2 Hz, 2H), 1.43-1.19 (m, 16H), 0.89 (t, J = 6.8 Hz, 3H), 0.83 (dt, J = 7.2, 2.0 Hz, 6H).

[0285] Example 23

[0286] [ka]

[0287] (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-(heptan-4-yloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl decanoate To a cold (ice-water bath) stirred solution of decanoic acid (44.6 mg, 0.259 mmol) in DCM (10 mL) was added N,N-dimethylpyridin-4-amine (132 mg, 1.080 mmol) and 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (207 mg, 1.080 mmol), and the resulting mixture was stirred for 0.5 h. Heptan-4-yl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (150 mg, 0.216 mmol) was then added, and the resulting mixture was stirred at 25 °C for 4 h. LCMS indicated the reaction was complete. The reaction mixture was partitioned between DCM (100 mL) and water (30 mL). The organic layer was washed with brine (20 mL), dried over NaSO, filtered, and concentrated. The residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 40 g, 0-100% CHCN / 10 mM NHHCO in water) to afford (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-(heptan-4-yloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl decanoate (150 mg, 0.175 mmol, 81% yield) as a colorless oil. LCMS (M+H)=849.3; Retention time (0.05%TFA)=2.28 minutes. HPLC retention time (10mM NH4HCO3) = 8.72 minutes. 1H NMR (400MHz, CDCl3) δ 8.00 (s, 1H), 7.23 (dd, J = 7.6, 2.4 Hz, 4H), 7.20-7.09 (m, 6H), 6.39 (t, J = 6.8 Hz, 1H), 6.00 (brs, 2H), 5.57-5.53 (m, 1H), 4.90-4.81 (m, 1H), 4.38-4.28 (m, 1H), 4.19 (dd, J = 11.2, 6.4 Hz, 1H), 4.05 (dd, J = 10.8, 5.6 Hz, 1H), 3.63 (t, J = 10.8 Hz, 1H), 3.07 (dd, J = 13.6, 6.4 Hz, 1H), 2.94 (dd, J = 14.4, 7.2 Hz, 1H), 2.62 (s, 1H), 2.61-2.55 (m, 2H), 2.40 (t, J = 7.6 Hz, 2H), 1.67 (quint, J = 7.2 Hz, 2H), 1.48-1.39 (m, 4H), 1.34-1.25 (m, 12H), 1.24-1.15 (m, 4H), 0.90-0.80 (m, 9H).

[0288] Example 24

[0289] [ka]

[0290] (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-(heptan-4-yloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl dodecanoate To a stirred solution of dodecanoic acid (47.6 mg, 0.238 mmol) in DCM (4 mL) was added DMAP (132 mg, 1.080 mmol) and EDC (207 mg, 1.080 mmol) at 0° C. After 0.5 h, a solution of heptan-4-yl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (150 mg, 0.216 mmol) in DCM (2 mL) was added. The resulting mixture was stirred at 15° C. overnight. LCMS indicated the reaction was complete. The reaction mixture was diluted with water (20 mL), the organic layer was separated, and the aqueous phase was extracted with DCM (20 mL × 2). The combined organic phase was washed with brine (20 mL), dried over NaSO, and concentrated in vacuo. The residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 12 g, 0-100% CHCN / 10 mM NH4HCO3 aqueous solution) to afford (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-(heptan-4-yloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl dodecanoate (103 mg, 0.116 mmol, 53.5% yield) as a white solid. LCMS (M+H)=877.7; Retention time (0.1%TFA)=2.837 minutes. HPLC: Retention time (10mM NH4HCO3) = 9.728 minutes. 1H NMR (400MHz, CDCl3) δ 8.03 (s, 1H), 7.25-7.21 (m, 4H), 7.19 (d, J = 7.0 Hz, 1H), 7.17-7.13 (m, 4H), 7.10 (t, J = 7.3 Hz, 1H), 6.39 (t, J = 6.8 Hz, 1H), 6.06 (s, 2H), 5.55 (t, J = 5.4 Hz, 1H), 4.88-4.81 (m, 1H), 4.33 (tt, J = 10.1, 6.7 Hz, 1H), 4.19 (dd, J = 11.1, 6.5 Hz, 1H), 4.05 (dd, J = 11.1, 5.8 Hz, 1H), 3.64 (t, J = 10.6 Hz, 1H), 3.07 (dd, J = 13.8, 5.9 Hz, 1H), 2.94 (dd, J = 13.8, 7.0 Hz, 1H), 2.62 (s, 1H), 2.57 (t, J = 6.3 Hz, 2H), 2.40 (t, J = 7.6 Hz, 2H), 1.71-1.62 (m, 2H), 1.47-1.38 (m, 4H), 1.33-1.24 (m, 16H), 1.18 (ddd, J = 11.7, 9.8, 6.1 Hz, 4H), 0.88 (t, J = 6.9 Hz, 3H), 0.83 (td, J = 7.3, 2.5 Hz, 6H).

[0291] Example 25

[0292] [ka]

[0293] (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-(heptan-4-yloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl tetradecanoate To a stirred solution of tetradecanoic acid (54.2 mg, 0.238 mmol) in DCM (4 mL) was added DMAP (132 mg, 1.080 mmol) and EDC (207 mg, 1.080 mmol) at 0° C. After 0.5 h, a solution of heptan-4-yl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (150 mg, 0.216 mmol) in DCM (2 mL) was added. The resulting mixture was stirred at 15° C. overnight. LCMS indicated the reaction was complete. The reaction mixture was diluted with water (20 mL), the organic layer was separated, and the aqueous phase was extracted with DCM (20 mL × 2). The combined organic phase was washed with brine (20 mL), dried over NaSO, and concentrated in vacuo. The crude product was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 12 g, 0-100% CHCN / 10 mM NH4HCO3 aqueous solution) to give (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-(heptan-4-yloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl tetradecanoate (140 mg, 0.153 mmol, 71.0% yield) as a white solid. LCMS (M+H)=905.1; Retention time (0.1%TFA)=3.237 minutes. HPLC: Retention time (10mM NH4HCO3) = 9.766 minutes. 1H NMR (400MHz, CDCl3) δ 8.05 (s, 1H), 7.26-7.21 (m, 4H), 7.20-7.17 (m, 1H), 7.15 (dd, J = 9.4, 4.9 Hz, 4H), 7.10 (t, J = 7.3 Hz, 1H), 6.39 (t, J = 6.7 Hz, 1H), 6.15 (s, 2H), 5.54 (t, J = 5.4 Hz, 1H), 4.89-4.81 (m, 1H), 4.33 (ddd, J = 16.6, 10.0, 6.8 Hz, 1H), 4.18 (dd, J = 11.1, 6.6 Hz, 1H), 4.06 (dd, J = 11.1, 5.8 Hz, 1H), 3.67 (s, 1H), 3.07 (dd, J = 13.7, 6.1 Hz, 1H), 2.94 (dd, J = 13.8, 7.1 Hz, 1H), 2.62 (s, 1H), 2.59-2.53 (m, 2H), 2.40 (t, J = 7.6 Hz, 2H), 1.66 (dd, J = 14.6, 7.2 Hz, 2H), 1.47-1.38 (m, 4H), 1.32 (s, 2H), 1.26 (s, 18H), 1.23-1.14 (m, 4H), 0.85 (ddd, J = 11.2, 9.6, 4.4 Hz, 9H).

[0294] Example 26

[0295] [ka]

[0296] (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-(heptan-4-yloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl palmitate To a cold (ice-water bath) stirred solution of palmitic acid (72.0 mg, 0.281 mmol) in DCM (10 mL) was added N,N-dimethylpyridin-4-amine (132 mg, 1.080 mmol) and 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (207 mg, 1.080 mmol), and the resulting mixture was stirred for 0.5 h. Heptan-4-yl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (150 mg, 0.216 mmol) was then added, and the resulting mixture was stirred at 25 °C for 4 h. LCMS indicated the reaction was complete. The reaction mixture was partitioned between DCM (80 mL) and water (20 mL). The organic layer was washed with brine (20 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 40 g, 0-100% MeOH / 10 mM NH4HCO3 in water) to afford (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-(heptan-4-yloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl palmitate (161 mg, 0.166 mmol, 77% yield) as a pale yellow semisolid. LCMS (M+H)=933.5; Retention time (0.05%TFA)=3.79 minutes. HPLC retention time (10mM NH4HCO3) = 12.41 minutes. 1H NMR (400MHz, CDCl3) δ 8.00 (s, 1H), 7.23 (dd, J = 7.6, 2.4 Hz, 4H), 7.21-7.18 (m, 1H), 7.18-7.13 (m, 4H), 7.10 (t, J = 7.2 Hz, 1H), 6.39 (t, J = 6.8 Hz, 1H), 6.01 (brs, 2H), 5.58-5.53 (m, 1H), 4.85 (quint, J = 6.0 Hz, 1H), 4.38-4.28 (m, 1H), 4.19 (dd, J = 11.2, 6.4 Hz, 1H), 4.05 (dd, J = 11.2, 5.6 Hz, 1H), 3.64 (t, J = 10.4 Hz, 1H), 3.07 (dd, J = 13.2, 5.6 Hz, 1H), 2.94 (dd, J = 14.0, 7.2 Hz, 1H), 2.62 (s, 1H), 2.61-2.55 (m, 2H), 2.40 (t, J = 7.6 Hz, 2H), 1.67 (quint, J = 6.8 Hz, 2H), 1.48-1.39 (m, 4H), 1.34-1.26 (m, 24H), 1.22-1.12 (m, 4H), 0.88 (t, J = 6.8 Hz, 3H), 0.83 (dt, J = 7.2, 2.8 Hz, 6H).

[0297] Example 27

[0298] [ka]

[0299] (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-(heptan-4-yloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl stearate To a cold (ice-water bath) stirred solution of stearic acid (133 mg, 0.468 mmol) in DCM (15 mL) was added N,N-dimethylpyridin-4-amine (220 mg, 1.799 mmol) and 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (345 mg, 1.799 mmol), and the resulting mixture was stirred for 0.5 h. Then, heptan-4-yl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (250 mg, 0.360 mmol) was added. The resulting mixture was stirred at 25° C. for 4 h. LCMS indicated the reaction was complete. The reaction mixture was partitioned between DCM (100 mL) and water (30 mL). The organic layer was washed with brine (20 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 40 g, 0-100% MeOH / 10 mM NH4HCO3 in water) to afford (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-(heptan-4-yloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl stearate (110 mg, 0.113 mmol, 31.3% yield) as a pale yellow solid. LCMS (M+H)=962.4; Retention time (0.05%TFA)=4.29 minutes. HPLC retention time (10mM NH4HCO3) = 17.03 minutes. 1H NMR (400MHz, CDCl3) δ 8.02 (s, 1H), 7.23 (dd, J = 7.6, 2.0 Hz, 4H), 7.21-7.18 (m, 1H), 7.18-7.13 (m, 4H), 7.10 (t, J = 7.2 Hz, 1H), 6.39 (t, J = 6.8 Hz, 1H), 5.97 (brs, 2H), 5.55 (t, J = 4.8 Hz, 1H), 4.85 (quint, J = 5.6 Hz, 1H), 4.38-4.29 (m, 1H), 4.19 (dd, J = 10.8, 6.4 Hz, 1H), 4.05 (dd, J = 11.2, 6.0 Hz, 1H), 3.61 (t, J = 10.4 Hz, 1H), 3.07 (dd, J = 13.6, 6.4 Hz, 1H), 2.94 (dd, J = 14.0, 7.2 Hz, 1H), 2.62 (s, 1H), 2.57 (t, J = 6.4 Hz, 2H), 2.40 (t, J = 7.2 Hz, 2H), 1.67 (quint, J = 7.2 Hz, 2H), 1.49-1.39 (m, 4H), 1.29-1.22 (m, 28H), 1.21-1.10 (m, 4H), 0.88 (t, J = 6.4 Hz, 3H), 0.83 (dt, J = 7.6, 2.8 Hz, 6H).

[0300] Example 28

[0301] [ka]

[0302] (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-(heptan-4-yloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl icosanoate To a stirred solution of icosanoic acid (82 mg, 0.261 mmol) in DCM (4 mL) was added DMAP (145 mg, 1.188 mmol) and EDC (228 mg, 1.188 mmol) at 0° C. After 0.5 h, a solution of heptan-4-yl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (165 mg, 0.238 mmol) in DCM (2 mL) was added. The resulting mixture was stirred overnight at 15° C. TLC (DCM:MeOH=20:1, R f =0.3) indicated the presence of a new compound. The reaction mixture was diluted with water (20 mL), the organic layer was separated, and the aqueous layer was extracted with DCM (20 mL × 2). The combined organic phases were washed with brine (30 mL), dried over Na2SO4, and concentrated in vacuo. The residue was analyzed by preparative TLC (DCM:MeOH = 20:1, R f =0.3) to afford (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-(heptan-4-yloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl icosanoate (143 mg, 0.138 mmol, 58.1% yield) as a white solid. HPLC: Retention time (10 mM NH4HCO3) = 13.594 min. 1H NMR (400MHz, CDCl3) δ 8.10 (s, 1H), 7.24 (d, J = 7.7 Hz, 4H), 7.19 (d, J = 7.0 Hz, 1H), 7.17 (d, J = 1.4 Hz, 1H), 7.14 (d, J = 8.4 Hz, 3H), 7.10 (t, J = 7.3 Hz, 1H), 6.38 (t, J = 6.7 Hz, 1H), 6.04 (s, 2H), 5.54 (t, J = 5.3 Hz, 1H), 4.86 (dd, J = 12.2, 5.8 Hz, 1H), 4.36-4.28 (m, 1H), 4.18 (dd, J = 11.1, 6.6 Hz, 1H), 4.05 (dd, J = 11.1, 6.0 Hz, 1H), 3.61 (t, J = 10.7 Hz, 1H), 3.08 (dd, J = 13.6, 5.7 Hz, 1H), 2.94 (dd, J = 13.8, 7.1 Hz, 1H), 2.62 (s, 1H), 2.59-2.52 (m, 2H), 2.40 (t, J = 7.6 Hz, 2H), 1.71-1.62 (m, 2H), 1.48-1.39 (m, 4H), 1.38-1.30 (m, 4H), 1.25 (s, 28H), 1.22-1.15 (m, 4H), 0.85 (ddd, J = 10.4, 9.6, 4.4 Hz, 9H).

[0303] Example 29

[0304] [ka]

[0305] (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-(heptan-4-yloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl docosanoate To a stirred solution of docosanoic acid (89 mg, 0.261 mmol) in DCM (4 mL) was added DMAP (145 mg, 1.188 mmol) and EDC (228 mg, 1.188 mmol) at 0° C. After 0.5 h, heptan-4-yl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (165 mg, 0.238 mmol) was added at 0° C. The resulting mixture was stirred at 15° C. for 16 h. TLC (DCM:MeOH=20:1, Rf=0.3) indicated the presence of a new compound. The reaction mixture was diluted with water (10 mL), the organic layer was separated, and the aqueous layer was extracted with DCM (20 mL × 2). The combined organic phases were washed with brine (30 mL), dried over NaSO, and concentrated in vacuo. The mixture was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 0-100% CHOH / 10 mM NH4HCO3 aqueous solution) to afford (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-(heptan-4-yloxy)-1-oxo-3-phenylpropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl docosanoate (107 mg, 0.104 mmol, 43.7% yield) as a white solid. HPLC: Retention time (10mM NH4HCO3) = 17.825 minutes. 1H NMR (400MHz, CDCl3) δ 8.01 (s, 1H), 7.25-7.20 (m, 4H), 7.18 (d, J = 7.0 Hz, 1H), 7.17-7.12 (m, 4H), 7.09 (t, J = 7.3 Hz, 1H), 6.39 (t, J = 6.7 Hz, 1H), 6.19 (s, 2H), 5.55 (t, J = 5.2 Hz, 1H), 4.84 (dd, J = 11.9, 6.2 Hz, 1H), 4.33 (t, J = 6.4 Hz, 1H), 4.19 (dd, J = 11.0, 6.5 Hz, 1H), 4.06 (dd, J = 11.0, 5.8 Hz, 1H), 3.71 (t, J = 10.6 Hz, 1H), 3.07 (dd, J = 13.7, 5.9 Hz, 1H), 2.94 (dd, J = 13.7, 6.9 Hz, 1H), 2.61 (s, 1H), 2.56 (d, J = 6.4 Hz, 2H), 2.39 (t, J = 7.5 Hz, 2H), 1.66 (dd, J = 14.4, 7.2 Hz, 2H), 1.46-1.38 (m, 4H), 1.30 (s, 2H), 1.25 (s, 34H), 1.21-1.14 (m, 4H), 0.84 (ddd, J = 11.4, 9.2, 4.2 Hz, 9H).

[0306] Example 30

[0307]

change

[0308] Project 1: ヘキシル(4-ニトロェニル)カーボネート To a mixture of 4-nitrophenyl carbonochloridate (230 g, 1141 mmol) and hexane-1-ol (117 g, 1141 mmol) in DCM (1000 mL) was added triethylamine (239 mL, 1712 mmol) and stirred at room temperature for 1 h. TLC showed the reaction was complete. The reaction mixture was concentrated to give crude hexyl(4-nitrophenyl)carbonate (280 g, 943 mmol, 103% yield) as a yellow oil, which was used in the next step without purification.

[0309] Step 2: Octyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((hexyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a stirred solution of octyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (160 mg, 0.226 mmol) in DCM (10 mL) was added triethylamine (0.094 mL, 0.677 mmol) followed by hexyl(4-nitrophenyl)carbonate (121 mg, 0.452 mmol) and stirred at room temperature for 2 days. TLC indicated the formation of a new product. The residue was concentrated and purified using flash chromatography (silica gel, 0–10% MeOH / DCM) to afford octyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((hexyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (160 mg, 0.187 mmol, 83% yield) as a white foam. LCMS (M+H) = 837.7; retention time (0.1% TFA) = 2.43 min. 1H NMR (400MHz, CDCl3) δ 7.26 - 7.05 (m, 10H), 6.38 (t, J = 6.7 Hz, 1H), 6.15 (s, 2H), 5.47 (dd, J = 6.6, 4.5 Hz, 1H), 4.47 - 4.25 (m, 2H), 4.27 - 4.12 (m, 3H), 4.02 (t, J = 6.7 Hz, 2H), 3.74 (t, J = 10.7 Hz, 1H), 3.00 (d, J = 6.2 Hz, 2H), 2.83 - 2.60 (m, 3H), 1.82 - 1.62 (m, 2H), 1.60 - 1.46 (m, 2H), 1.38 (dd, J = 14.2, 6.3 Hz, 2H), 1.36 - 1.15 (m, 14H). 0.89 (dt, J = 13.8, 6.9 Hz, 6H).

[0310] Example 31

[0311] [ka]

[0312] Step 1: Decyl ((S)-(perfluorophenoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a stirred solution of decyl L-phenylalaninate (3 g, 9.82 mmol) in DCM (20 mL) was added triethylamine (1.502 mmol, 10.80 mmol) dropwise at -70 °C. To this mixture was added a solution of phenyl phosphorodichloridate (2.051 g, 9.72 mmol) in anhydrous DCM (8 mL) dropwise. The reaction mixture was stirred at this temperature for 30 min, then warmed to 0 °C over 2 h and stirred for 1 h. To this mixture was added a solution of 2,3,4,5,6-pentafluorophenol (1.790 g, 9.72 mmol) and triethylamine (1.502 mL, 10.80 mmol) in DCM (30 mL) dropwise and stirred at 0 °C for 2 h. LCMS indicated the reaction was complete. The white solid (triethylamine hydrochloride) was filtered off and washed with DCM (10 mL). The filtrate was concentrated under reduced pressure, and the residue was triturated with tert-butyl methyl ether (20 mL), and triethylamine hydrochloride was removed by filtration. The cake was washed with tert-butyl methyl ether (2 × 10 mL), and the combined filtrates were concentrated under reduced pressure. The resulting residue was triturated with 10% EtOAc / hexane (30 mL), and the solid was collected by filtration to give decyl ((S)-(perfluorophenoxy)(phenoxy)phosphoryl)-L-phenylalaninate (1.5 g, 2.390 mmol, 24.34% yield) as a white solid. 31 P-NMR: >98% de (LCMS (M+H) = 628.0; retention time (0.1% TFA) = 2.78 min). 31 P NMR (162MHz, DMSO-d6) δ 0.22.

[0313] Step 2: Decyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a stirred solution of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (0.2 g, 0.682 mmol) in THF (10 mL) and pyridine (2 mL) was added tert-butylmagnesium chloride (1.432 mL, 1.432 mmol) dropwise at 0 °C and stirred at 0 °C for 30 min. A solution of decyl ((S)-(perfluorophenoxy)(phenoxy)phosphoryl)-L-phenylalaninate (0.428 g, 0.682 mmol) in THF (25 mL) was added dropwise to the above solution at -15 °C and stirred at -15 °C for 16 h. TLC indicated the presence of a new compound. The reaction was quenched with 2 M HCl (10 mL) and extracted with EtOAc (50 mL × 3). The combined organic phase was washed with NaHCO3 solution (50 mL), brine (50 mL), dried over Na2SO4, filtered, and concentrated in vacuo. The crude product was purified using flash chromatography (silica gel, 0-10% MeOH / DCM) to afford decyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (160 mg, 0.217 mmol, 31.8% yield) as a white solid. 1 H NMR (400MHz, CDCl3) δ 7.88 (s, 1H), 7.28 (d, J = 8.0 Hz, 2H), 7.25 - 7.07 (m, 8H), 6.31 (dd, J = 7.4, 4.3 Hz, 1H), 6.06 - 5.72 (m, 2H), 4.71 (d, J = 5.3 Hz, 1H), 4.37 - 4.12 (m, 3H), 4.04 (t, J = 6.8 Hz, 2H), 3.71 (s, 1H), 3.47 (d, J = 5.2 Hz, 1H), 3.00 (d, J = 6.3 Hz, 2H), 2.81 - 2.55 (m, 3H), 1.53 (s, 2H), 1.26 (d, J = 23.2 Hz, 14H), 0.88 (t, J = 6.8 Hz, 3H).

[0314] Step 3: Decyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((hexyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a stirred mixture of decyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (140 mg, 0.190 mmol) in DCM (10 mL) was added triethylamine (0.079 mL, 0.570 mmol), followed by hexyl(4-nitrophenyl)carbonate (102 mg, 0.380 mmol), and the mixture was stirred at room temperature for 2 days. TLC showed the presence of a new compound. The reaction mixture was concentrated and purified using flash chromatography (silica gel, 0–10% MeOH / DCM) to afford decyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((hexyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (130 mg, 0.150 mmol, 79% yield) as a white foam. LCMS (M+H) = 865.7; retention time (0.1% TFA) = 2.60 min. 1H NMR (400MHz, CDCl3) δ 7.94 (s, 1H), 7.26 - 7.07 (m, 10H), 6.38 (t, J = 6.7 Hz, 1H), 6.05 - 5.64 (m, 2H), 5.48 (dd, J = 6.9, 4.4 Hz, 1H), 4.31 (ddd, J = 13.6, 10.4, 5.0 Hz, 2H), 4.17 (dt, J = 16.5, 5.8 Hz, 3H), 4.01 (t, J = 6.8 Hz, 2H), 3.63 (t, J = 10.7 Hz, 1H), 3.00 (d, J = 6.3 Hz, 2H), 2.83 - 2.62 (m, 3H), 1.76 -1.69 (m, 2H), 1.57 - 1.47 (m, 2H), 1.38 (dd, J = 15.0, 6.4 Hz, 2H), 1.35 - 1.21 (m, 18H), 0.89 (dt, J = 13.8, 6.9Hz, 6H).

[0315] Example 32

[0316] [ka]

[0317] Dodecyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((hexyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a solution of dodecyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (252 mg, 0.329 mmol) in DCM (10 mL) was added triethylamine (0.138 mL, 0.988 mmol) and DMAP (40.3 mg, 0.329 mmol), followed by hexyl(4-nitrophenyl)carbonate (176 mg, 0.659 mmol) and stirred at room temperature for 2 days. LCMS showed the reaction was complete. The reaction mixture was concentrated and purified using silica gel chromatography (0-5% MeOH / DCM) to afford dodecyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((hexyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (120 mg, 0.131 mmol, 39.8% yield) as a white solid. LCMS (M+H) = 892.7; retention time (0.1% TFA) = 2.517 min. 1 H NMR (400MHz, CDCl3) δ 8.01 (s, 1H), 7.26 - 7.04 (m, 10H), 6.37 (d, J = 6.4 Hz, 1H), 6.17 (s, 2H), 5.47 (s, 1H), 4.31 (d, J = 7.8 Hz, 2H), 4.18 (d, J = 6.1 Hz, 3H), 4.01 (d, J = 6.4 Hz, 2H), 3.73 (s, 1H), 3.00 (s, 2H), 2.69 (d, J = 18.1 Hz, 3H), 1.69 (d, J = 6.9 Hz, 2H), 1.51 (s, 2H), 1.46 - 1.07 (m, 24H), 0.89 (d, J = 8.0 Hz, 6H).

[0318] Example 33

[0319] [ka]

[0320] Tetradecyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((hexyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a stirred mixture of tetradecyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (150 mg, 0.189 mmol) in DCM (10 mL) was added triethylamine (0.026 mL, 0.189 mmol) and DMAP (23.11 mg, 0.189 mmol), followed by hexyl(4-nitrophenyl)carbonate (101 mg, 0.378 mmol), and the reaction mixture was stirred at 23° C. for 2 days. The reaction mixture was concentrated, and the residue was purified using flash chromatography (40 g silica column, 0–10% MeOH / DCM) to afford tetradecyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((hexyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (125 mg, 0.135 mmol, 71.4% yield) as a white solid. LCMS (M+H) = 920.7; retention time (0.1% TFA) = 2.849 min. 1H NMR (400MHz, CDCl3) δ 8.04 (s, 1H), 7.26 - 7.07 (m, 10H), 6.38 (t, J = 6.7 Hz, 1H), 6.16 (s, 2H), 5.47 (dd, J = 6.6, 4.5 Hz, 1H), 4.38 - 4.24 (m, 2H), 4.24 - 4.09 (m, 3H), 4.02 (t, J = 6.7 Hz, 2H), 3.75 (t, J = 10.7 Hz, 1H), 3.13 - 2.89 (m, 2H), 2.81 - 2.61 (m, 3H), 1.82 - 1.62 (m, 2H), 1.61 - 1.46 (m, 2H), 1.46 - 1.35 (m, 2H), 1.33 - 1.18 (m, 26H), 0.89 (dt, J = 10.1, 6.9 Hz, 6H).

[0321] Example 34

[0322]

change

[0323] Project 1: Tert-ブトキシカルボニル-L-フェニルアラニネート To a stirred mixture of (tert-butoxycarbonyl)-L-phenylalanine (20 g, 75 mmol), HOBt (15.28 g, 113 mmol), and EDC (17.34 g, 90 mmol) in DCM (150 mL) was added TEA (10.51 mL, 75 mmol) at 0 °C. After 30 min, hexadecan-1-ol (18.28 g, 75 mmol) was added, and the resulting reaction mixture was stirred at 25 °C for 16 h. TLC showed the reaction was complete. The reaction mixture was diluted with water (80 mL), the organic layer was separated, and the aqueous layer was extracted with DCM (100 mL × 2). The combined organic layers were washed with brine, dried over Na2SO4, and concentrated. The residue was purified using silica gel chromatography (petroleum ether: EtOAc = 100:0 to 10:1) to give hexadecyl (tert-butoxycarbonyl)-L-phenylalaninate (17.2 g, 35.1 mmol, 46.6% yield) as a white solid. 1 H NMR (400MHz, CDCl3) δ 7.31 - 7.27 (m, 2H), 7.26 - 7.07 (m, 3H), 4.98 (d, J = 8.1 Hz, 1H), 4.57 (d, J = 7.0 Hz, 1H), 4.08 (dd, J = 6.9, 5.7 Hz, 2H), 3.19 - 2.85 (m, 2H), 1.59 (s, 2H), 1.42 (s, 9H), 1.25 (d, J = 6.6 Hz, 22H), 0.88 (t, J = 6.8 Hz, 3H).

[0324] Step 2: Hexadecyl L-phenylalaninate To a stirred solution of hexadecyl (tert-butoxycarbonyl)-L-phenylalaninate (17 g, 34.7 mmol) in DCM (75 mL) was added TFA (30 mL, 389 mmol) at 0 °C. After 5 min, the mixture was stirred at 15 °C for 2 h. TLC showed the presence of a new compound. The reaction mixture was concentrated, and the residue was diluted with DCM (100 mL), and the pH was adjusted to 8–9 with saturated Na2CO3. The organic layer was separated, and the aqueous layer was extracted with DCM (2 × 100 mL). The combined organic layers were washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified using silica gel chromatography (DCM:MeOH = 20:1) to give hexadecyl L-phenylalaninate (13 g, 33.4 mmol, 86% yield). 1 H NMR (400MHz, CDCl3) δ 7.30 (t, J = 7.2 Hz, 2H), 7.26 - 7.17 (m, 3H), 4.09 (t, J = 6.7 Hz, 2H), 3.72 (dd, J = 7.8, 5.4 Hz, 1H), 3.08 (dd, J = 13.5, 5.4 Hz, 1H), 2.87 (dd, J = 13.5, 7.8 Hz, 1H), 1.59 (d, J = 6.8 Hz, 2H), 1.27 (d, J = 9.3 Hz, 28H), 0.88 (t, J = 6.8 Hz, 3H).

[0325] Step 3: Hexadecyl ((S)-(perfluorophenoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a stirred solution of hexadecyl L-phenylalaninate (10 g, 25.7 mmol) in anhydrous DCM (100 mL) was added triethylamine (3.94 mL, 28.2 mmol) dropwise over 15 minutes at -70°C. To this mixture was added a solution of phenylphosphonic dichloride (5.00 g, 25.7 mmol) in anhydrous DCM (50 mL) over 1 hour. The reaction mixture was stirred at this temperature for an additional 30 minutes, then warmed to 0°C over 2 hours and stirred for 1 hour. To this mixture was added a solution of 2,3,4,5,6-pentafluorophenol (4.72 g, 25.7 mmol) and TEA (3.94 mL, 28.2 mmol) in DCM (30 mL) over 20 minutes and stirred at 0°C for 16 hours. TLC indicated the reaction was complete. The white solid (triethylamine hydrochloride) was filtered off and washed with DCM (50 mL). The filtrate was concentrated under reduced pressure, the residue was triturated with tert-butyl methyl ether (150 mL), and the triethylamine hydrochloride was removed by filtration. The cake was washed with tert-butyl methyl ether (2 x 100 mL), and the combined filtrates were concentrated under reduced pressure to give 12 g of crude solid containing an equal mixture of diastereomers. The mixture was triturated with 20% EtOAc / hexane (100 mL), and the solid was collected by filtration to give hexadecyl ((S)-(perfluorophenoxy)(phenoxy)phosphoryl)-L-phenylalaninate (5.2 g, 6.94 mmol, 27.0% yield) as a white solid. 31 >98% de as determined by P-NMR). 1H NMR (400MHz, CDCl3) δ 7.35 (t, J = 7.8 Hz, 2H), 7.26 - 7.13 (m, 8H), 7.13 - 6.93 (m, 2H), 4.44 (ddd, J = 15.6, 10.1, 5.8 Hz, 1H), 4.07 (ddt, J = 10.7, 6.7, 5.6 Hz, 2H), 3.79 (t, J = 11.1 Hz, 1H), 3.19 - 2.96 (m, 3H), 1.70 (d, J = 11.5 Hz, 2H), 1.40 (dd, J = 15.5, 8.2 Hz, 1H), 1.35 - 1.20 (m, 26H), 0.91 - 0.82 (m, 3H). 31 P NMR (162MHz, CDCl3) δ -1.57.

[0326] Step 4: Hexadecyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a stirred solution of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (0.3 g, 1.023 mmol) in THF (20 mL) and pyridine (1.0 mL) was added tert-butylmagnesium chloride (2.148 mL, 2.148 mmol) dropwise at 0° C. The reaction was then stirred at 25° C. for 30 minutes. A solution of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (0.3 g, 1.023 mmol) in THF (20 mL) was added dropwise to the above solution at −15° C. and stirred at −15° C. for 12 hours. LCMS indicated the reaction was complete. The reaction was quenched with 2M HCl (10 mL) and extracted with EtOAc (20 mL × 3). The combined organic layers were washed with NaHCO3 solution (20 mL), washed with brine (20 mL), dried over Na2SO4, filtered, and concentrated in vacuo. The crude product was purified using silica gel chromatography (25 g, DCM:MeOH = 20:1) to give hexadecyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (301 mg, 0.348 mmol, 34.0% yield). 1 H NMR (400MHz, CDCl3)δ 7.90 (s, 1H), 7.25 - 7.01 (m, 10H), 6.31 (dd, J = 7.3, 4.3 Hz, 1H), 6.02 (s, 1H), 4.72 (t, J = 7.4 Hz, 1H), 4.23 (dt, J = 20.5, 8.5 Hz, 3H), 4.03 (t, J = 6.5 Hz, 2H), 3.00 (d, J = 5.9 Hz, 2H), 2.83 - 2.53 (m, 2H), 1.56 (d, J = 27.4 Hz, 3H), 1.35 - 1.14 (m, 28H), 0.88 (t, J = 6.7 Hz, 3H).

[0327] Step 5: Hexadecyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((hexyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a stirred mixture of hexadecyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (300 mg, 0.365 mmol) in DCM (20 mL) was added triethylamine (0.153 mL, 1.096 mmol), DMAP (44.6 mg, 0.365 mmol), and hexyl(4-nitrophenyl)carbonate (195 mg, 0.731 mmol) under N at 15 °C and stirred at 20 °C for 2 days. LCMS showed the presence of a new product. The reaction mixture was concentrated, and the residue was purified using flash chromatography (silica gel, 0-10% MeOH / DCM) to give hexadecyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((hexyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (120 mg, 0.124 mmol, 34.0% yield). LCMS (M+H) = 948.7; retention time (0.1% TFA) = 3.80 min. 1H NMR (400MHz, CDCl3) δ 7.95 (s, 1H), 7.27 - 7.03 (m, 10H), 6.48 - 6.26 (m, 1H), 6.01 (s, 1H), 5.48 (dd, J = 6.8, 4.4 Hz, 1H), 4.46 - 4.23 (m, 2H), 4.25 - 4.11 (m, 3H), 4.01 (t, J = 6.7 Hz, 2H), 3.70 (t, J = 10.7 Hz, 1H), 3.02 (t, J = 10.6 Hz, 2H), 2.80 - 2.61 (m, 3H), 1.75 - 1.63 (m, 2H), 1.36 - 1.17 (m, 34H), 0.89 (dt, J = 10.3, 6.9 Hz, 6H).

[0328] Example 35

[0329] [ka]

[0330] Step 1: Octadecyl (tert-butoxycarbonyl)-L-phenylalaninate To a stirred mixture of (tert-butoxycarbonyl)-L-phenylalanine (20 g, 75 mmol), HOBt (15.28 g, 113 mmol), and EDC (17.34 g, 90 mmol) in DCM (150 mL) was added TEA (10.51 mL, 75 mmol) at 0 °C. After 30 min, octadecan-1-ol (20.39 g, 75 mmol) was added, and the resulting reaction mixture was stirred at 25 °C for 16 h. TLC showed the reaction was complete. The reaction mixture was diluted with water (40 mL), the organic layer was separated, and the aqueous layer was extracted with DCM (50 mL × 2). The combined organic layers were washed with brine, dried over Na2SO4, and concentrated. The residue was purified using silica gel chromatography (petroleum ether: EtOAc = 100:0 to 10:1) to give octadecyl (tert-butoxycarbonyl)-L-phenylalaninate (20.2 g, 39.0 mmol, 51.8% yield) as a yellow oil. 1H NMR (400MHz, CDCl3) δ 7.32 - 7.27 (m, 2H), 7.26 - 7.11 (m, 3H), 4.98 (d, J = 8.1 Hz, 1H), 4.57 (d, J = 7.0 Hz, 1H), 4.12 - 4.04 (m, 2H), 3.14 - 3.00 (m, 2H), 1.65 - 1.53 (m, 4H), 1.42 (s, 9H), 1.29 - 1.22 (m, 28H), 0.88 (t, J = 6.8 Hz, 3H).

[0331] Step 2: Octadecyl L-phenylalaninate To a cold (ice-water bath) stirred solution of octadecyl (tert-butoxycarbonyl)-L-phenylalaninate (20 g, 38.6 mmol) in DCM (150 mL) was added TFA (50 mL, 649 mmol). After 5 min, the mixture was stirred at room temperature for 2 h. TLC indicated the presence of a new compound. The reaction mixture was concentrated, and the residue was diluted with DCM (100 mL) and the pH was adjusted to 8–9 with saturated Na2CO3. The organic layer was separated, and the aqueous layer was extracted with DCM (2 × 100 mL). The combined organic layers were washed with brine (80 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified using silica gel chromatography (DCM:MeOH = 20:1) to give octadecyl L-phenylalaninate (14.6 g, 35.0 mmol, 53.9% yield). 1 H NMR (400MHz, CDCl3) δ 7.30 (t, J = 7.2 Hz, 2H), 7.26 - 7.17 (m, 3H), 4.09 (t, J = 6.7 Hz, 2H), 3.72 (dd, J = 7.8, 5.4 Hz, 1H), 3.08 (dd, J = 13.5, 5.4 Hz, 1H), 2.87 (dd, J = 13.5, 7.8 Hz, 1H), 1.59 (d, J = 6.8 Hz, 2H), 1.27 (d, J = 9.3 Hz, 30H), 0.88 (t, J = 6.8 Hz, 3H).

[0332] Step 3: Octadecyl ((S)-(perfluorophenoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a stirred solution of octadecyl L-phenylalaninate (14.1 g, 33.8 mmol) in anhydrous DCM (200 mL) was added triethylamine (5.16 mL, 37.1 mmol) dropwise over 15 minutes at -70°C. To this mixture was added a solution of phenyl phosphorodichloridate (7.05 g, 33.4 mmol) in anhydrous DCM (50 mL) over 1 hour. The reaction mixture was stirred at this temperature for an additional 30 minutes, then warmed to 0°C over 2 hours and stirred for 1 hour. To this mixture was added a solution of 2,3,4,5,6-pentafluorophenol (6.15 g, 33.4 mmol) and triethylamine (5.16 mL, 37.1 mmol) in DCM (50 mL) over 20 minutes and stirred at 0°C for 4 hours. TLC indicated the reaction was complete. The white solid (triethylamine hydrochloride) was filtered off and washed with DCM (50 mL). The filtrate was concentrated under reduced pressure, and then the residue was triturated with tert-butyl methyl ether (500 mL). Triethylamine hydrochloride was removed by filtration. The cake was washed with tert-butyl methyl ether (2 x 50 mL), and the combined filtrates were concentrated under reduced pressure to give 25 g of crude solid containing an equal mixture of diastereomers. The mixture was triturated with 20% EtOAc / hexane (400 mL), and the solid was collected by filtration to give octadecyl ((S)-(perfluorophenoxy)(phenoxy)phosphoryl)-L-phenylalaninate (10 g, 13.52 mmol, 40.0% yield) as a white solid. 31 >98% de:N76107-96-A1) as determined by P-NMR. 1H NMR (400MHz, CDCl3) δ 7.35 (t, J = 7.8 Hz, 2H), 7.26 - 7.13 (m, 6H), 7.05 (d, J = 7.4 Hz, 2H), 4.44 (s, 1H), 4.13 - 3.95 (m, 2H), 3.77 (t, J = 11.1 Hz, 1H), 3.10 (ddd, J = 39.1, 13.8, 6.0 Hz, 2H), 1.54 (s, 2H), 1.26 (s, 30H), 0.88 (t, J = 6.7 Hz, 3H). 31 P NMR (162MHz, CDCl3) δ -1.57.

[0333] Step 4: Octadecyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a stirred solution of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (0.3 g, 1.023 mmol) in THF (40 mL) and pyridine (2.000 mL) was added tert-butylmagnesium chloride (2.148 mL, 2.148 mmol) dropwise at 0°C, and the reaction was stirred at 25°C for 30 min. A solution of octadecyl ((S)-(perfluorophenoxy)(phenoxy)phosphoryl)-L-phenylalaninate (0.984 g, 1.330 mmol) in THF (20 mL) was added dropwise to the above solution at -15°C, and the mixture was stirred at -15°C for 4 h. TLC showed the presence of a new compound. The reaction mixture was quenched with 2M HCl (20 mL) and extracted with EtOAc (30 mL × 3). The combined organic layers were washed with NaHCO3 solution (20 mL), brine (20 mL), dried over Na2SO4, filtered, and concentrated in vacuo. The crude product was purified using silica gel chromatography (0–10% MeOH / DCM) to afford octadecyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (320 mg, 0.362 mmol, 35.4% yield) as a white solid. 1 H NMR (400MHz, CDCl3) δ 7.90 (s, 1H), 7.26 - 7.04 (m, 10H), 6.31 (dd, J = 7.1, 4.4 Hz, 1H), 6.28 - 5.93 (m, 2H), 4.71 (s, 1H), 4.35 - 4.15 (m, 3H), 4.02 (t, J = 6.7 Hz, 2H), 3.95 (t, J = 11.0 Hz, 1H), 3.82 (s, 1H), 2.99 (d, J = 6.2 Hz, 2H), 2.81 - 2.61 (m, 3H), 1.51 (s, 2H), 1.24 (d, J = 9.7 Hz, 30H), 0.88 (t, J = 6.8 Hz, 3H).

[0334] Step 5: Octadecyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((hexyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a stirred mixture of octadecyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (320 mg, 0.377 mmol) in DCM (20 mL) was added triethylamine (0.158 mL, 1.131 mmol), DMAP (46.0 mg, 0.377 mmol), and hexyl(4-nitrophenyl)carbonate (403 mg, 1.508 mmol) under N at 15° C. The mixture was stirred at room temperature for 3 days. LCMS showed the reaction was complete. The reaction mixture was concentrated in vacuo, and the residue was purified using silica gel chromatography (25 g, 0-7% MeOH / DCM) to afford octadecyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((hexyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (191 mg, 0.188 mmol, 49.8% yield) as a pale yellow solid. LCMS; retention time (0.1% TFA) = 4.08 min. 1H NMR (400MHz, CDCl3) δ 7.95 (s, 1H), 7.26 - 7.00 (m, 10H), 6.92 - 6.44 (m, 2H), 6.37 (s, 1H), 5.48 (d, J = 2.5 Hz, 1H), 4.30 (dd, J = 11.2, 6.5 Hz, 2H), 4.24 - 4.07 (m, 4H), 4.01 (t, J = 6.4 Hz, 2H), 3.11 - 2.90 (m, 2H), 2.67 (s, 3H), 1.78 - 1.65 (m, 2H), 1.51 (s, 2H), 1.44 - 1.15 (m, 36H), 0.89 (dt, J = 10.3, 6.9 Hz, 6H).

[0335] Example 36

[0336] [ka]

[0337] Step 1: Eicosyl (tert-butoxycarbonyl)-L-phenylalaninate To a stirred mixture of (tert-butoxycarbonyl)-L-phenylalanine (20 g, 75 mmol), HOBt (15.28 g, 113 mmol), and EDC (17.34 g, 90 mmol) in DCM (150 mL) was added TEA (10.51 mL, 75 mmol) at 0 °C. After 30 min, icosan-1-ol (22.51 g, 75 mmol) was added, and the resulting reaction mixture was stirred at 25 °C for 16 h. TLC showed the reaction was complete. The reaction mixture was diluted with water (80 mL), the organic layer was separated, and the aqueous layer was extracted with DCM (100 mL × 2). The combined organic layers were washed with brine, dried over Na2SO4, and concentrated. The residue was purified using silica gel chromatography (petroleum ether: EtOAc = 100:0 to 10:1) to give icosyl (tert-butoxycarbonyl)-L-phenylalaninate (20 g, 36.6 mmol, 48.6% yield) as a white solid. 1H NMR (400MHz, CDCl3) δ 7.33 - 7.27 (m, 2H), 7.25 - 7.08 (m, 3H), 4.98 (d, J = 8.2 Hz, 1H), 4.57 (d, J = 7.5 Hz, 1H), 4.07 (tt, J = 5.4, 2.7 Hz, 2H), 3.09 (dd, J = 13.9, 7.3 Hz, 2H), 1.57 - 1.52 (m, 2H), 1.42 (s, 9H), 1.26 (s, 34H), 0.88 (t, J = 6.8 Hz, 3H).

[0338] Step 2: Eicosyl L-phenylalaninate To a cold (ice-water bath) stirred solution of icosyl (tert-butoxycarbonyl)-L-phenylalaninate (19.5 g, 35.7 mmol) in DCM (150 mL) was added 2,2,2-trifluoroacetic acid. After 5 min, the mixture was stirred at 25 °C for 2 h. TLC indicated the presence of a new compound. The reaction mixture was concentrated, and the residue was diluted with DCM (100 mL) and the pH was adjusted to 8-9 with saturated Na2CO3. The organic layer was separated, and the aqueous layer was extracted with DCM (2 × 100 mL). The combined organic layers were washed with brine (80 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified using silica gel chromatography (DCM:MeOH = 20:1) to give icosyl L-phenylalaninate (14.5 g, 32.5 mmol, 91% yield). 1 H NMR (400MHz, CDCl3) δ 7.30 (t, J = 7.2 Hz, 2H), 7.26 - 7.17 (m, 3H), 4.09 (t, J = 6.7 Hz, 2H), 3.72 (dd, J = 7.8, 5.4 Hz, 1H), 3.08 (dd, J = 13.5, 5.4 Hz, 1H), 2.87 (dd, J = 13.5, 7.8 Hz, 1H), 1.59 (d, J = 6.8 Hz, 2H), 1.27 (d, J = 9.3 Hz, 34H), 0.88 (t, J = 6.8 Hz, 3H).

[0339] Step 3: Eicosyl ((S)-(perfluorophenoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a stirred solution of icosyl L-phenylalaninate (12.6 g, 28.3 mmol) in anhydrous DCM (100 mL) was added triethylamine (4.32 mL, 31.1 mmol) dropwise over 15 minutes at -70°C. To this mixture was added a solution of phenyl phosphorodichloridate (5.90 g, 28.0 mmol) in anhydrous DCM (35 mL) over 1 hour. The reaction mixture was stirred at this temperature for an additional 30 minutes, then warmed to 0°C over 2 hours and stirred for 1 hour. To this mixture was added a solution of 2,3,4,5,6-pentafluorophenol (5.15 g, 28.0 mmol) and triethylamine (4.32 mL, 31.1 mmol) in DCM (30 mL) over 20 minutes and stirred at 0°C for 4 hours. TLC indicated the reaction was complete. The white solid (triethylamine hydrochloride) was filtered off and washed with DCM (50 mL). The filtrate was concentrated under reduced pressure, the residue was triturated with tert-butyl methyl ether (250 mL), and the triethylamine hydrochloride was removed by filtration. The cake was washed with tert-butyl methyl ether (2 x 50 mL), and the combined filtrates were concentrated under reduced pressure to give 26 g of crude solid containing an equal mixture of diastereomers. The mixture was triturated with 20% EtOAc / hexane (200 mL), and the solid was collected by filtration to give icosyl ((S)-(perfluorophenoxy)(phenoxy)phosphoryl)-L-phenylalaninate (5 g, 6.51 mmol, 23.04% yield) as a white solid. 31 >98% de as determined by P-NMR). 1H NMR (400MHz, CDCl3) δ 7.35 (t, J = 7.8 Hz, 2H), 7.26 - 7.13 (m, 6H), 7.05 (d, J = 7.4 Hz, 2H), 4.44 (s, 1H), 4.13 - 3.95 (m, 2H), 3.77 (t, J = 11.1 Hz, 1H), 3.10 (ddd, J = 39.1, 13.8, 6.0 Hz, 2H), 1.54 (s, 2H), 1.26 (s, 34H), 0.88 (t, J = 6.7 Hz, 3H). 31 P NMR (162MHz, CDCl3) δ -1.57.

[0340] Step 4: Eicosyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a stirred solution of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (0.3 g, 1.023 mmol) in THF (40 mL) and pyridine (2.000 mL) was added tert-butylmagnesium chloride (2.148 mL, 2.148 mmol) dropwise at 0°C. The reaction was then stirred at 25°C for 30 minutes. A solution of icosyl ((S)-(perfluorophenoxy)(phenoxy)phosphoryl)-L-phenylalaninate (1.257 g, 1.637 mmol) in THF (20 mL) was added dropwise to the above solution at -15°C and stirred at -15°C for 4 hours. TLC indicated the presence of a new compound. The reaction was quenched with 2M HCl (20 mL) and extracted with EtOAc (50 mL × 3). The combined organic layers were washed with NaHCO3 solution (20 mL), brine (20 mL), dried over Na2SO4, filtered, and concentrated in vacuo. The crude product was purified using silica gel chromatography (DCM / MeOH = 100 / 0 to 10 / 1) to afford icosyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (330 mg, 0.365 mmol, 35.6% yield) as a white solid. 1 H NMR (400MHz, CDCl3) δ 7.90 (s, 1H), 7.26 - 7.04 (m, 10H), 6.31 (dd, J = 7.1, 4.4 Hz, 1H), 6.28 - 5.93 (m, 2H), 4.71 (s, 1H), 4.35 - 4.15 (m, 3H), 4.02 (t, J = 6.7 Hz, 2H), 3.95 (t, J = 11.0 Hz, 1H), 3.82 (s, 1H), 2.99 (d, J = 6.2 Hz, 2H), 2.81 - 2.61 (m, 3H), 1.51 (s, 2H), 1.24 (d, J = 9.7 Hz, 34H), 0.88 (t, J = 6.8 Hz, 3H).

[0341] Step 5: Icosyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((hexyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a stirred mixture of icosyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (330 mg, 0.376 mmol) in DCM (20 mL) was added triethylamine (0.157 mL, 1.129 mmol), DMAP (46.0 mg, 0.376 mmol), and hexyl(4-nitrophenyl)carbonate (402 mg, 1.505 mmol) under N at 15° C. The mixture was stirred at 15° C. for 3 days. TLC showed the reaction was complete. The reaction mixture was concentrated in vacuo, and the residue was purified using silica gel chromatography (25 g, 0–7% MeOH / DCM) to afford icosyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((hexyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (150 mg, 0.149 mmol, 39.7% yield) as a white solid. HPLC: RT (10 mM NH4HCO3) = 22.56 min. 1 H NMR: N78913-5-A1. 1 H NMR (400MHz, CDCl3) δ 7.97 (s, 1H), 7.26 - 7.04 (m, 10H), 6.38 (s, 1H), 6.32 - 5.98 (m, 2H), 5.47 (s, 1H), 4.31 (s, 2H), 4.19 (s, 3H), 4.02 (s, 2H), 3.77 (s, 1H), 3.01 (s, 2H), 2.67 (s, 3H), 1.69 (d, J = 6.8 Hz, 2H), 1.52 (s, 2H), 1.45 - 1.16 (m, 40H), 0.95 - 0.83 (m, 6H).

[0342] Example 37

[0343] [ka]

[0344] Step 1: Docosyl (tert-butoxycarbonyl)-L-phenylalaninate A mixture of docosan-1-ol (19.70 g, 60.3 mmol), (tert-butoxycarbonyl)-L-phenylalanine (16 g, 60.3 mmol), EDC (17.34 g, 90 mmol), imidazole (12.32 g, 181 mmol), DIPEA (31.6 mL, 181 mmol), and (tert-butoxycarbonyl)-L-phenylalanine (16 g, 60.3 mmol) in DCM (500 mL) was stirred at 25 °C for 16 h. TLC indicated the presence of a new compound. The reaction mixture was diluted with water (500 mL), the organic layer was separated, and the aqueous layer was extracted with DCM (250 mL × 3). The combined organic phases were washed with brine, dried over Na2SO4, filtered, and concentrated. The residue was purified using silica gel chromatography (petroleum ether: EtOAc = 20:1) to give docosyl (tert-butoxycarbonyl)-L-phenylalaninate (12 g, 20.91 mmol, 34.7% yield) as a white solid. H NMR: N76106-76-A1. 1 H NMR (400MHz, CDCl3) δ 7.33 - 7.06 (m, 5H), 4.98 (d, J = 7.9 Hz, 1H), 4.57 (d, J = 7.0 Hz, 1H), 4.17 - 3.94 (m, 2H), 3.15 - 2.99 (m, 2H), 1.65 - 1.56 (m, 2H), 1.42 (s, 9H), 1.26 (s, 38H), 0.88 (t, J = 6.8 Hz, 3H).

[0345] Step 2: Docosyl L-phenylalaninate To a cold (ice-water bath) stirred solution of docosyl (tert-butoxycarbonyl)-L-phenylalaninate (12 g, 20.91 mmol) in DCM (500 mL) was added TFA (16.11 mL, 209 mL). After 5 min, the mixture was stirred at room temperature for 2 h. TLC showed the presence of a new compound. The reaction mixture was washed with water and saturated Na2CO3. The combined aqueous layer was extracted with DCM (400 mL × 3). The combined organic layer was washed with brine, dried over Na2SO4, filtered, and concentrated. The residue was purified using silica gel chromatography (DCM:MeOH = 20:1) to give docosyl L-phenylalaninate (10 g, 19.00 mmol, 91% yield). 1 H NMR (400MHz, CDCl3) δ 7.43 - 6.96 (m, 5H), 4.09 (t, J = 6.7 Hz, 2H), 3.72 (dd, J = 7.8, 5.4 Hz, 1H), 3.08 (dd, J = 13.5, 5.4 Hz, 1H), 2.87 (dd, J = 13.5, 7.8 Hz, 1H), 1.71 - 1.51 (m, 4H), 1.26 (s, 38H), 0.88 (t, J = 6.8 Hz, 3H).

[0346] Step 3: Docosyl ((S)-(perfluorophenoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a stirred solution of docosyl L-phenylalaninate (16 g, 33.8 mmol) in anhydrous DCM (100 mL) was added triethylamine (5.16 mL, 37.1 mmol) dropwise over 15 min at -70 °C. To this mixture was added a solution of phenyl phosphorodichloridate (7.05 g, 33.4 mmol) in anhydrous DCM (35 mL) over 1 h. The reaction mixture was stirred at this temperature for an additional 30 min, then warmed to 0 °C over 2 h and stirred for 1 h. To this mixture was added a solution of 2,3,4,5,6-pentafluorophenol (6.15 g, 33.4 mmol) and triethylamine (5.16 mL, 37.1 mmol) in DCM (30 mL) over 20 min and stirred at 0 °C for 4 h. TLC indicated the reaction was complete. The white solid (triethylamine hydrochloride) was filtered off and washed with DCM (50 mL). The filtrate was concentrated under reduced pressure, the residue was triturated with tert-butyl methyl ether (250 mL), and triethylamine hydrochloride was removed by filtration. The cake was washed with tert-butyl methyl ether (2 × 50 mL), and the combined filtrates were concentrated under reduced pressure to give 21 g of a crude solid containing an equal mixture of diastereomers. The mixture was triturated with 20% EtOAc / hexanes (200 mL), and the solid was collected by filtration to give docosyl ((S)-(perfluorophenoxy)(phenoxy)phosphoryl)-L-phenylalaninate (5.5 g, 6.91 mmol, 20.46% yield) as a white solid ( 31 >98% de:N76107-98-A1) as determined by P-NMR. 1 H NMR (400MHz, CDCl3) δ 7.35 (t, J = 7.8 Hz, 2H), 7.26 - 7.13 (m, 6H), 7.05 (d, J = 7.4 Hz, 2H), 4.44 (s, 1H), 4.13 - 3.95 (m, 2H), 3.77 (t, J = 11.1 Hz, 1H), 3.10 (ddd, J = 39.1, 13.8, 6.0 Hz, 2H), 1.54 (s, 2H), 1.26 (s, 38H), 0.88 (t, J = 6.7 Hz, 3H). 31 P NMR (162MHz, CDCl3) δ -1.57.

[0347] Step 4: Docosyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a stirred solution of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (0.3 g, 1.023 mmol) in THF (40 mL) and pyridine (2.0 mL) was added tert-butylmagnesium chloride (2.148 mL, 2.148 mmol) dropwise at 0°C. The reaction was then stirred at 25°C for 30 minutes. A solution of docosyl ((S)-(perfluorophenoxy)(phenoxy)phosphoryl)-L-phenylalaninate (1.628 g, 2.046 mmol) in THF (20 mL) was added dropwise to the above solution at -15°C and stirred at -15°C for 4 hours. TLC indicated the presence of a new compound. The reaction was quenched with 2M HCl (20 mL) and extracted with EtOAc (30 mL × 3). The combined organic layers were washed with NaHCO3 solution (20 mL), brine (20 mL), dried over Na2SO4, filtered, and concentrated in vacuo. The crude product was purified using column chromatography (0–10% MeOH / DCM) to afford docosyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (340 mg, 0.362 mmol, 35.4% yield) as a white solid. 1H NMR (400MHz, CDCl3) δ 7.90 (s, 1H), 7.26 - 7.06 (m, 10H), 6.31 (dd, J = 7.1, 4.4 Hz, 1H), 6.26 - 5.83 (m, 2H), 4.72 (s, 1H), 4.35 - 4.17 (m, 3H), 4.07 - 3.95 (m, 3H), 3.80 (s, 1H), 2.99 (d, J = 6.2 Hz, 2H), 2.83 - 2.55 (m, 3H), 1.51 (s, 2H), 1.36 - 1.04 (m, 38H), 0.88 (t, J = 6.8 Hz, 3H).

[0348] Step 5: Docosyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((hexyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate To a stirred mixture of docosyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (340 mg, 0.376 mmol) in DCM (20 mL) was added triethylamine (0.157 mL, 1.127 mmol), DMAP (45.9 mg, 0.376 mmol), and hexyl(4-nitrophenyl)carbonate (402 mg, 1.503 mmol) under N at 15° C. The mixture was stirred at room temperature for 3 days. TLC showed the reaction was complete. The reaction was concentrated in vacuo and the residue was purified using silica gel chromatography (25 g, 0-7% MeOH / DCM) to afford docosyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((hexyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-phenylalaninate (166 mg, 0.161 mmol, 42.8% yield) as a white solid. HPLC: RT (10 mM NH4HCO3) = 17.516 min. 1 H NMR: N78913-6-A1. 1 H NMR (400MHz, CDCl3) δ 7.95 (s, 1H), 7.27 - 7.02 (m, 10H), 6.38 (t, J = 6.7 Hz, 3H), 5.60 - 5.40 (m, 1H), 4.30 (dd, J = 10.9, 6.4 Hz, 2H), 4.19 (t, J = 6.4 Hz, 3H), 4.01 (t, J = 6.7 Hz, 3H), 3.08 - 2.91 (m, 2H), 2.70 (d, J = 30.0 Hz, 3H), 1.80 - 1.66 (m, 2H), 1.51 (s, 2H), 1.43 - 1.10 (m, 44H), 0.89 (dt, J = 10.2, 6.8 Hz, 6H).

[0349] Example 38

[0350] [ka]

[0351] Step 1: Isopropyl ((S)-(perfluorophenoxy)(phenoxy)phosphoryl)-L-alaninate (N76107-86) To a stirred solution of isopropyl L-alaninate hydrochloride (8.0 g, 47.7 mmol) in anhydrous dichloromethane (50 mL) was added TEA (13.77 mL, 99 mmol) dropwise over 15 minutes at -70°C. To this mixture was added a solution of phenylphosphonic acid dichloride (9.21 g, 47.2 mmol) in anhydrous dichloromethane (50 mL) over 1 hour. The reaction mixture was stirred at this temperature for an additional 30 minutes, then warmed to 0°C over 2 hours and stirred for 1 hour. To this mixture was added a solution of 2,3,4,5,6-pentafluorophenol (8.70 g, 47.2 mmol) and TEA (7.32 mL, 52.5 mmol) in dichloromethane (30 mL) over 20 minutes. The crude mixture was stirred at 0°C for 16 hours. LCMS indicated the reaction was complete. The white solid (triethylamine hydrochloride) was filtered off and washed with dichloromethane (25 mL). The filtrate was concentrated under reduced pressure, the residue was triturated with TBME (150 mL), and the triethylamine hydrochloride was removed by filtration. The cake was washed with TBME (2 x 25 mL), and the combined filtrates were concentrated under reduced pressure to give 22 g of crude solid containing an equal mixture of diastereomers. The mixture was triturated with 20% EtOAc in hexane (100 mL) and collected by filtration to give isopropyl ((S)-(perfluorophenoxy)(phenoxy)phosphoryl)-L-alaninate (7.0 g, 15.44 mmol, 32.4% yield) as a white solid ( 31 >98% de as determined by P-NMR). 1H NMR (400MHz, CDCl3) δ 7.36 (t, J = 7.9 Hz, 2H), 7.26 - 7.17 (m, 3H), 5.04 (dt, J = 12.5, 6.3 Hz, 1H), 4.14 (dd, J = 16.1, 8.3 Hz, 1H), 4.08 - 3.87 (m, 1H), 1.46 (d, J = 7.0 Hz, 3H), 1.25 (t, J = 5.9 Hz, 6H). 31 P NMR (162MHz, CDCl3) δ -1.60.

[0352] Step 2: Isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (N78870-36) To a solution of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (0.25 g, 0.852 mmol) in THF (50 mL) and pyridine (2.50 mL), tert-butylmagnesium chloride (1.705 mL, 1.705 mmol) was added dropwise at −15° C. and stirred for 1 hour at −15° C. Next, a solution of isopropyl ((S)-(perfluorophenoxy)(phenoxy)phosphoryl)-L-alaninate (0.502 g, 1.108 mmol) in THF (15 mL) was added to the reaction mixture, and the resulting reaction mixture was stirred at −15° C. for 2 hours. LCMS showed a product / by-product ratio of 1:1. The reaction mixture was quenched with NH4Cl (4 mL) and partitioned between EtOAc (200 mL) and water (30 mL). The organic layer was washed with brine (20 mL), dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified using silica gel column chromatography (8% MeOH in DCM) to afford isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (90 mg, 0.154 mmol, 18.08% yield) as a white solid. LCMS (M+H) = 563.3; retention time (0.05% TFA) = 1.44 min.

[0353] Step 3: (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl nonanoate To a solution of nonanoic acid (50.6 mg, 0.320 mmol) in DCM (8 mL) were added N,N-dimethylpyridin-4-amine (32.6 mg, 0.267 mmol) and 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (102 mg, 0.533 mmol), and the resulting mixture was stirred at ice-water bath temperature for 0.5 h. Then, isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (150 mg, 0.267 mmol) was added, and the resulting mixture was stirred at 25 °C for 16 h. LCMS showed the reaction was complete. The reaction mixture was concentrated, and the residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 40 g, 0-100% CH3CN / 10 mM NH4HCO3 in water) to afford (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl nonanoate (127 mg, 0.177 mmol, 66.2% yield) as a white solid. LCMS (M+H) = 703.0; retention time (10 mM NH4HCO3) = 1.80 min. 1H NMR (400MHz, CDCl3) δ 8.06 (s, 1H), 7.31-7.27 (m, 2H), 7.21 (d, J = 8.8 Hz, 2H), 7.13 (t, J = 7.2 Hz, 1H), 6.43 (t, J = 6.4 Hz, 1H), 6.05 (brs, 2H), 5.65 (dd, J = 7.2, 4.4 Hz, 1H), 5.01 (quint, J = 6.4 Hz, 1H), 4.43 (dd, J = 11.2, 6.0 Hz, 1H), 4.35 (dd, J = 11.2, 5.6 Hz, 1H), 4.06-3.95 (m, 1H), 3.84 (dd, J = 11.2, 9.6 Hz, 1H), 2.72 (quint, J = 7.2 Hz, 1H), 2.64 (s, 1H), 2.64-2.59 (m, 1H), 2.40 (t, J = 7.6 Hz, 2H), 1.67 (quint, J = 7.6 Hz, 2H), 1.36 (d, J = 6.8 Hz, 3H), 1.35-1.25 (m, 10H), 1.22 (d, J = 6.0 Hz, 6H), 0.89 (t, J = 6.8 Hz, 3H)

[0354] Example 39

[0355] [ka]

[0356] (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl decanoate To a solution of decanoic acid (59.7 mg, 0.347 mmol) in DCM (8 mL) were added N,N-dimethylpyridin-4-amine (32.6 mg, 0.267 mmol) and 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (102 mg, 0.533 mmol), and the resulting mixture was stirred at ice-water bath temperature for 0.5 h. Then, isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (150 mg, 0.267 mmol) was added, and the resulting mixture was stirred at 25 °C for 16 h. LCMS indicated the reaction was complete. The reaction mixture was concentrated, and the residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 40 g, 0-100% CH3CN / 10 mM NH4HCO3 in water) to afford (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl decanoate (125 mg, 0.172 mmol, 64.5% yield) as a white solid. LCMS (M+H) = 717.5; retention time (0.05% TFA) = 2.00 min. 1H NMR (400MHz, CDCl3) δ 8.05 (s, 1H), 7.28 (t, J = 8.4 Hz, 2H), 7.20 (d, J = 8.8 Hz, 2H), 7.13 (t, J = 7.6 Hz, 1H), 6.43 (t, J = 6.8 Hz, 1H), 6.04 (brs, 2H), 5.65 (dd, J = 6.8, 4.0 Hz, 1H), 5.01 (quint, J = 6.4 Hz, 1H), 4.43 (dd, J = 10.8, 6.4 Hz, 1H), 4.35 (dd, J = 10.8, 6.0 Hz, 1H), 4.06-3.95 (m, 1H), 3.82 (dd, J = 11.2, 9.6 Hz, 1H), 2.76-2.67 (m, 1H), 2.66-2.59 (m, 2H), 2.40 (t, J = 7.6 Hz, 2H), 1.67 (quint, J = 7.6 Hz, 2H), 1.36 (d, J = 7.2 Hz, 3H), 1.33-1.24 (m, 12H), 1.22 (d, J = 6.4 Hz, 6H), 0.88 (t, J = 6.8 Hz, 3H).

[0357] Example 40

[0358] [ka]

[0359] (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl dodecanoate To a solution of dodecanoic acid (64.1 mg, 0.320 mmol) in DCM (8 mL) were added N,N-dimethylpyridin-4-amine (32.6 mg, 0.267 mmol) and 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (102 mg, 0.533 mmol), and the resulting mixture was stirred at ice-water bath temperature for 0.5 h. Then, isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (150 mg, 0.267 mmol) was added, and the resulting mixture was stirred at 25 °C for 16 h. LCMS indicated the reaction was complete. The reaction mixture was concentrated, and the residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 40 g, 0-100% CH3CN / 10 mM NH4HCO3 in water) to afford (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl dodecanoate (79 mg, 0.102 mmol, 38.3% yield) as a white solid. LCMS (M+H) = 745.5; retention time (0.05% TFA) = 2.13 min. 1H NMR (400MHz, CDCl3) δ 8.05 (s, 1H), 7.28 (t, J = 8.4 Hz, 2H), 7.21 (d, J = 8.8 Hz, 2H), 7.13 (t, J = 7.2 Hz, 1H), 6.43 (t, J = 6.8 Hz, 1H), 5.94 (brs, 2H), 5.65 (dd, J = 6.8, 4.0 Hz, 1H), 5.01 (quint, J = 6.4 Hz, 1H), 4.43 (dd, J = 11.2, 6.4 Hz, 1H), 4.35 (dd, J = 11.2, 6.0 Hz, 1H), 4.06-3.95 (m, 1H), 3.79 (dd, J = 11.2, 9.6 Hz, 1H), 2.72 (quint, J = 6.8 Hz, 1H), 2.64 (s, 1H), 2.63-2.59 (m, 1H), 2.40 (t, J = 7.6 Hz, 2H), 1.66-1.62 (m, 2H), 1.36 (d, J = 6.8 Hz, 3H), 1.34-1.24 (m, 16H), 1.22 (d, J = 6.4 Hz, 6H), 0.88 (t, J = 6.8 Hz, 3H).

[0360] Example 41

[0361] [ka]

[0362] (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl tetradecanoate To a solution of tetradecanoic acid (73.1 mg, 0.320 mmol) in DCM (8 mL) were added N,N-dimethylpyridin-4-amine (32.6 mg, 0.267 mmol) and 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (102 mg, 0.533 mmol), and the resulting mixture was stirred at ice-water bath temperature for 0.5 h. Then, isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (150 mg, 0.267 mmol) was added, and the resulting mixture was stirred at 25 °C for 16 h. LCMS indicated the reaction was complete. The reaction mixture was concentrated, and the residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 40 g, 0-100% CH3CN / 10 mM NH4HCO3 in water) to afford (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl tetradecanoate (115 mg, 0.147 mmol, 55.3% yield) as a white solid. LCMS (M+H) = 773.5; retention time (0.05% TFA) = 2.30 min. 1H NMR (400MHz, CDCl3) δ 8.06 (s, 1H), 7.31-7.27 (m, 2H), 7.21 (d, J = 8.4 Hz, 2H), 7.13 (t, J = 7.2 Hz, 1H), 6.43 (t, J = 6.8 Hz, 1H), 6.06 (brs, 2H), 5.65 (dd, J = 6.8, 4.0 Hz, 1H), 5.01 (quint, J = 6.4 Hz, 1H), 4.43 (dd, J = 10.8, 6.0 Hz, 1H), 4.35 (dd, J = 10.8, 5.6 Hz, 1H), 4.05-3.96 (m, 1H), 3.84 (dd, J = 11.2, 9.6 Hz, 1H), 2.72 (quint, J = 6.8 Hz, 1H), 2.64 (s, 1H), 2.64-2.59 (m, 1H), 2.40 (t, J = 7.6 Hz, 2H), 1.67 (quint, J = 7.2 Hz, 2H), 1.36 (d, J = 7.2 Hz, 3H), 1.33-1.25 (m, 20H), 1.22 (d, J = 6.4 Hz, 6H), 0.88 (t, J = 6.8 Hz, 3H).

[0363] Example 42

[0364] [ka]

[0365] (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl palmitate To a solution of palmitic acid (82 mg, 0.320 mmol) in DCM (8 mL) were added N,N-dimethylpyrilzin-4-amine (32.6 mg, 0.267 mmol) and 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (102 mg, 0.533 mmol), and the resulting mixture was stirred at ice-water bath temperature for 0.5 h. Then, isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (150 mg, 0.267 mmol) was added, and the resulting mixture was stirred at 25 °C for 16 h. LCMS indicated the reaction was complete. The reaction mixture was concentrated, and the residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 40 g, 0-100% CH3CN / 10 mM NH4HCO3 in water) to afford (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl palmitate (114 mg, 0.141 mmol, 52.9% yield) as a white solid. LCMS (M+H) = 801.5; retention time (0.05% TFA) = 2.53 min. 1H NMR (400MHz, CDCl3) δ 8.06 (s, 1H), 7.31-7.27 (m, 2H), 7.21 (d, J = 8.4 Hz, 2H), 7.13 (t, J = 7.2 Hz, 1H), 6.43 (t, J = 6.8 Hz, 1H), 6.14 (brs, 2H), 5.65 (dd, J = 6.8, 4.0 Hz, 1H), 5.01 (quint, J = 6.0 Hz, 1H), 4.43 (dd, J = 10.8, 6.0 Hz, 1H), 4.35 (dd, J = 11.2, 6.0 Hz, 1H), 4.06-3.96 (m, 1H), 3.87 (dd, J = 11.2, 9.6 Hz, 1H), 2.71 (quint, J = 6.8 Hz, 1H), 2.66-2.59 (m, 2H), 2.40 (t, J = 7.6 Hz, 2H), 1.66 (quint, J = 7.2 Hz, 2H), 1.38 (d, J = 6.8 Hz, 3H), 1.32-1.25 (m, 24H), 1.21 (d, J = 6.4 Hz, 6H), 0.88 (t, J = 6.8 Hz, 3H).

[0366] Example 43

[0367] [ka]

[0368] (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl stearate To a solution of stearic acid (182 mg, 0.640 mmol) in DCM (8 mL) were added N,N-dimethylpyridin-4-amine (65.2 mg, 0.533 mmol) and 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (204 mg, 1.067 mmol), and the resulting mixture was stirred at ice-water bath temperature for 0.5 h. Then, isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (300 mg, 0.533 mmol) was added, and the resulting mixture was stirred at 25 °C for 16 h. LCMS indicated the reaction was complete. The reaction mixture was concentrated, and the residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 40 g, 0-100% CH3CN / 10 mM NH4HCO3 in water) to afford (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl stearate (126 mg, 0.147 mmol, 27.6% yield) as a white solid. LCMS (M+H) = 829.4; retention time (0.05% TFA) = 2.83 min. 1H NMR (400MHz, CDCl3) δ 8.07 (s, 1H), 7.31-7.27 (m, 2H), 7.21 (d, J = 8.8 Hz, 2H), 7.13 (t, J = 7.2 Hz, 1H), 6.44 (t, J = 6.8 Hz, 1H), 6.09 (brs, 2H), 5.65 (dd, J = 6.8, 4.0 Hz, 1H), 5.01 (quint, J = 6.4 Hz, 1H), 4.43 (dd, J = 10.8, 6.0 Hz, 1H), 4.35 (dd, J = 11.2, 6.0 Hz, 1H), 4.06-3.96 (m, 1H), 3.84 (dd, J = 11.2, 9.6 Hz, 1H), 2.71 (quint, J = 6.8 Hz, 1H), 2.66-2.59 (m, 2H), 2.40 (t, J = 7.6 Hz, 2H), 1.66 (quint, J = 7.2 Hz, 2H), 1.37 (d, J = 7.2 Hz, 3H), 1.32-1.25 (m, 28H), 1.22 (d, J = 6.0 Hz, 6H), 0.88 (t, J = 6.8 Hz, 3H).

[0369] Example 44

[0370] [ka]

[0371] (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl 2-propylpentanoate To a solution of 2-propylpentanoic acid (46.1 mg, 0.320 mmol) in DCM (8 mL) were added N,N-dimethylpyridin-4-amine (32.6 mg, 0.267 mmol) and 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (102 mg, 0.533 mmol), and the resulting mixture was stirred at ice-water bath temperature for 0.5 h. Then, isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (150 mg, 0.267 mmol) was added, and the resulting mixture was stirred at 25 °C for 16 h. LCMS indicated the reaction was complete. The reaction mixture was concentrated, and the residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 40 g, 0-100% CH3CN / 10 mM NH4HCO3 in water) to afford (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-((((S)-(((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)tetrahydrofuran-3-yl 2-propylpentanoate (105 mg, 0.150 mmol, 56.1% yield) as a white solid. LCMS (M+H) = 689.0; retention time (10 mM NH4HCO3) = 1.71 min. 1H NMR (400MHz, CDCl3) δ 8.04 (s, 1H), 7.31-7.27 (m, 2H), 7.20 (d, J = 8.8 Hz, 2H), 7.13 (t, J = 7.2 Hz, 1H), 6.42 (t, J = 6.4 Hz, 1H), 5.87 (brs, 2H), 5.65 (dd, J = 6.8, 4.8 Hz, 1H), 5.00 (quint, J = 6.4 Hz, 1H), 4.44 (dd, J = 11.2, 6.0 Hz, 1H), 4.33 (dd, J = 11.2, 5.6 Hz, 1H), 4.05-3.96 (m, 1H), 3.79 (dd, J = 11.2, 9.6 Hz, 1H), 2.80 (quint, J = 6.8 Hz, 1H), 2.68-2.60 (m, 2H), 2.51-2.43 (m, 1H), 1.71-1.65 (m, 2H), 1.52-1.44 (m, 2H), 1.36 (d, J = 7.2 Hz, 3H), 1.35-1.27 (m, 4H), 1.21 (d, J = 6.0 Hz, 6H), 0.91 (t, J = 7.2 Hz, 6H).

[0372] Example 45

[0373] [ka]

[0374] Isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((hexyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate A mixture of isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (180 mg, 0.320 mmol) and DMAP (39.1 mg, 0.320 mmol) in DCM (6 mL) was treated with TEA (0.134 mL, 0.960 mmol) followed by hexyl(4-nitrophenyl)carbonate (428 mg, 1.600 mmol), and the mixture was stirred at 25° C. for 2 days. LCMS showed the reaction was complete. The reaction mixture was concentrated, and the residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 40 g, 0-100% CH3CN / 10 mM NH4HCO3 in water) to give isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((hexyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (105 mg, 0.145 mmol, 45.4% yield) as a white solid. LCMS (M+H) = 691.3; retention time (0.05% TFA) = 1.81 min. 1H NMR (400MHz, CDCl3) δ 8.01 (s, 1H), 7.31-7.27 (m, 2H), 7.20 (d, J = 8.4 Hz, 2H), 7.13 (t, J = 7.2 Hz, 1H), 6.42 (t, J = 6.4 Hz, 1H), 5.97 (brs, 2H), 5.57 (dd, J = 7.2, 4.4 Hz, 1H), 5.00 (quint, J = 6.4 Hz, 1H), 4.46 (dd, J = 11.2, 6.4 Hz, 1H), 4.37 (dd, J = 11.2, 6.4 Hz, 1H), 4.19 (dt, J = 6.8, 2.8 Hz, 2H), 4.06-3.95 (m, 1H), 3.83 (dd, J = 11.2, 9.6 Hz, 1H), 2.85 (quint, J = 7.2 Hz, 1H), 2.76-2.70 (m, 1H), 2.69 (s, 1H), 1.70 (quint, J = 6.8 Hz, 2H), 1.36 (d, J = 7.2 Hz, 3H), 1.36-1.24 (m, 6H), 1.21 (d, J = 6.4 Hz, 6H), 0.90 (t, J = 6.8 Hz, 3H).

[0375] Example 46

[0376]

change

[0377] Project 1: ヘプチル(4-ニトロェニル)カーボネート To a mixture of heptan-1-ol (4 g, 34.4 mmol) and DMAP (0.421 g, 3.44 mmol) in DCM (80 mL) was added TEA (7.20 mL, 51.6 mmol) dropwise at 0 °C. After stirring for 15 min, 4-nitrophenyl carbonochloridate (8.16 g, 34.4 mmol) was added. The resulting reaction mixture was stirred at 25 °C for 16 h. TLC showed the reaction was complete. The reaction mixture was partitioned between DCM (100 mL) and water (50 mL). The organic layer was washed with brine (40 mL), dried over Na2SO4, filtered, and concentrated to give heptyl (4-nitrophenyl)carbonate (9 g, 28.8 mmol, 84% yield) as a yellow oil. 1 H NMR (400MHz, CDCl3) δ 8.28 (dd, J = 7.2, 2.4 Hz, 2H), 7.39 (dd, J = 7.2, 2.0 Hz, 2H), 4.29 (t, J = 6.8 Hz, 2H), 1.76 (quint, J = 6.8 Hz, 2H), 1.46-1.27 (m, 8H), 0.90 (t, J = 6.8 Hz, 3H).

[0378] Step 2: Isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((heptyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate A mixture of isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (200 mg, 0.356 mmol) and DMAP (43.4 mg, 0.356 mmol) in DCM (6 mL) was treated with TEA (0.149 mL, 1.067 mmol) followed by heptyl (4-nitrophenyl)carbonate (500 mg, 1.778 mmol), and the mixture was stirred at 25° C. for 2 days. LCMS showed the reaction was complete. The reaction mixture was concentrated, and the residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 40 g, 0-100% CH3CN / 10 mM NH4HCO3 in water) to give isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((heptyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (97 mg, 0.135 mmol, 38.0% yield) as a white solid. LCMS (M+H) = 705.4; retention time (0.05% TFA) = 1.87 min. 1H NMR (400MHz, CDCl3) δ 8.06 (s, 1H), 7.28 (t, J = 8.0 Hz, 2H), 7.20 (d, J = 8.4 Hz, 2H), 7.12 (t, J = 6.8 Hz, 1H), 6.42 (t, J = 6.8 Hz, 1H), 5.97 (brs, 2H), 5.56 (dd, J = 6.8, 4.0 Hz, 1H), 5.00 (quint, J = 6.4 Hz, 1H), 4.46 (dd, J = 10.8, 6.4 Hz, 1H), 4.37 (dd, J = 11.2, 6.0 Hz, 1H), 4.19 (dt, J = 6.8, 2.4 Hz, 2H), 4.06-3.95 (m, 1H), 3.79 (dd, J = 11.2, 9.6 Hz, 1H), 2.83 (quint, J = 6.8 Hz, 1H), 2.76-2.70 (m, 1H), 2.69 (s, 1H), 1.70 (quint, J = 6.8 Hz, 2H), 1.36 (d, J = 6.8 Hz, 3H), 1.36-1.25 (m, 8H), 1.22 (d, J = 6.0 Hz, 6H), 0.89 (t, J = 6.8 Hz, 3H).

[0379] Example 47

[0380]

change

[0381] Engineering 1:4-Dynamic Flow Controller To a mixture of octan-1-ol (4 g, 30.7 mmol) and DMAP (0.375 g, 3.07 mmol) in DCM (120 mL) was added TEA (4.66 g, 46.1 mmol) dropwise at 0 °C. After stirring for 15 min, 4-nitrophenyl carbonochloridate (6.19 g, 30.7 mmol) was added. The resulting reaction mixture was stirred at 25 °C for 16 h. TLC indicated the reaction was complete. The reaction mixture was partitioned between DCM (120 mL) and water (50 mL), and the organic layer was washed with brine (40 mL), dried over Na2SO4, filtered, and concentrated to give 4-nitrophenyl octyl carbonate (9 g, 27.4 mmol, 89% yield) as a yellow oil. 1 H NMR (400MHz, CDCl3) δ 8.28 (dd, J = 7.2, 2.4 Hz, 2H), 7.38 (dd, J = 7.2, 2.0 Hz, 2H), 4.29 (t, J = 6.8 Hz, 2H), 1.76 (quint, J = 6.8 Hz, 2H), 1.45-1.24 (m, 10H), 0.89 (t, J = 6.8 Hz, 3H).

[0382] Step 2: Isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((octyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate A mixture of isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (200 mg, 0.356 mmol) and DMAP (43.4 mg, 0.356 mmol) in DCM (6 mL) was treated with TEA (0.149 mL, 1.067 mmol) followed by 4-nitrophenyloctyl carbonate (525 mg, 1.778 mmol), and the mixture was stirred at 25° C. for 2 days. LCMS showed the reaction was complete. The reaction mixture was concentrated, and the residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 40 g, 0-100% CH3CN / 10 mM NH4HCO3 in water) to give isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((octyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (110 mg, 0.150 mmol, 42.1% yield) as a white solid. LCMS (M+H) = 719.2; retention time (0.05% TFA) = 1.92 min. 1H NMR (400MHz, CDCl3) δ 8.01 (s, 1H), 7.31-7.27 (m, 2H), 7.22-7.16 (m, 2H), 7.13 (t, J = 7.2 Hz, 1H), 6.42 (t, J = 6.8 Hz, 1H), 6.00 (brs, 2H), 5.56 (dd, J = 7.2, 4.4 Hz, 1H), 5.00 (quint, J = 6.4 Hz, 1H), 4.46 (dd, J = 11.2, 6.4 Hz, 1H), 4.37 (dd, J = 11.2, 6.0 Hz, 1H), 4.19 (dt, J = 6.8, 2.4 Hz, 2H), 4.06-3.97 (m, 1H), 3.82 (dd, J = 11.2, 9.6 Hz, 1H), 2.85 (quint, J = 6.8 Hz, 1H), 2.76-2.70 (m, 1H), 2.69 (s, 1H), 1.70 (quint, J = 7.2 Hz, 2H), 1.36 (d, J = 7.2 Hz, 3H), 1.34-1.24 (m, 10H), 1.21 (d, J = 6.0 Hz, 6H), 0.89 (t, J = 6.8 Hz, 3H).

[0383] Example 48

[0384]

change

[0385] Engineering 1:4-Dynamic pipe A solution of 4-nitrophenyl carbonochloridate (5 g, 24.81 mmol), nonan-1-ol (3.58 g, 24.81 mmol), and DMAP (0.303 g, 2.481 mmol) in DCM (100 mL) was treated with TEA (5.19 mL, 37.2 mmol), and the mixture was stirred at room temperature for 3 h. TLC showed the reaction was complete. Water (80 mL) was added, and the mixture was extracted with DCM (100 mL × 2). The combined organic phase was washed with brine, dried over Na2SO4, and concentrated to give 4-nitrophenyl nonyl carbonate (5 g, 13.74 mmol, 55.4% yield) as a yellow oil. 1 H NMR (400MHz, CDCl3) δ 8.39 - 8.22 (m, 2H), 7.46 - 7.31 (m, 2H), 4.29 (t, J = 6.7 Hz, 2H), 1.88 - 1.72 (m, 2H), 1.30 (d, J = 11.3 Hz, 12H), 0.90 - 0.85 (m, 3H).

[0386] Step 2: Isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((nonyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate A mixture of isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (200 mg, 0.356 mmol) and DMAP (43.4 mg, 0.356 mmol) in DCM (5 mL) was treated with TEA (0.149 mL, 1.067 mmol) followed by 4-nitrophenyl nonyl carbonate (550 mg, 1.778 mmol), and the mixture was stirred at 25° C. for 2 days. LCMS showed the reaction was complete. The reaction mixture was concentrated, and the residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 40 g, 0-100% CH3CN / 10 mM NH4HCO3 in water) to give isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((nonyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (115 mg, 0.150 mmol, 42.1% yield) as a white solid. LCMS (M+H) = 733.3; retention time (0.05% TFA) = 1.98 min. 1H NMR (400MHz, CDCl3) δ 8.01 (s, 1H), 7.31-7.27 (m, 2H), 7.22-7.18 (m, 2H), 7.12 (t, J = 7.2 Hz, 1H), 6.42 (t, J = 6.8 Hz, 1H), 6.00 (brs, 2H), 5.56 (dd, J = 6.8, 4.0 Hz, 1H), 5.00 (quint, J = 6.4 Hz, 1H), 4.46 (dd, J = 11.2, 6.4 Hz, 1H), 4.37 (dd, J = 11.2, 6.0 Hz, 1H), 4.19 (dt, J = 6.8, 2.4 Hz, 2H), 4.07-3.96 (m, 1H), 3.83 (dd, J = 11.2, 9.6 Hz, 1H), 2.85 (quint, J = 6.8 Hz, 1H), 2.76-2.70 (m, 1H), 2.68 (s, 1H), 1.70 (quint, J = 7.2 Hz, 2H), 1.36 (d, J = 7.2 Hz, 3H), 1.34-1.25 (m, 12H), 1.21 (d, J = 6.0 Hz, 6H), 0.88 (t, J = 6.8 Hz, 3H).

[0387] Example 49

[0388]

change

[0389] Project 1: Detail A solution of 4-nitrophenyl carbonochloridate (5 g, 24.81 mmol), decan-1-ol (3.93 g, 24.81 mmol), and DMAP (0.303 g, 2.481 mmol) in DCM (100 mL) was treated with TEA (5.19 mL, 37.2 mmol), and the mixture was stirred at room temperature for 3 h. TLC showed the reaction was complete. Water (80 mL) was added, and the mixture was extracted with DCM (100 mL × 2). The combined organic phase was washed with brine, dried over Na2SO4, and concentrated to give decyl (4-nitrophenyl)carbonate (5.1 g, 13.40 mmol, 54.0% yield) as a yellow oil. 1 H NMR (400MHz, CDCl3) δ 8.39 - 8.22 (m, 2H), 7.46 - 7.31 (m, 2H), 4.29 (t, J = 6.7 Hz, 2H), 1.88 - 1.72 (m, 2H), 1.30 (d, J = 11.3 Hz, 14H), 0.90 - 0.85 (m, 3H).

[0390] Step 2: Isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-(((decyloxy)carbonyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate A mixture of isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (200 mg, 0.356 mmol) and DMAP (43.4 mg, 0.356 mmol) in DCM (5 mL) was treated with TEA (0.149 mL, 1.067 mmol) followed by decyl (4-nitrophenyl) carbonate (575 mg, 1.778 mmol), and the mixture was stirred at 25° C. for 2 days. LCMS showed the reaction was complete. The reaction mixture was concentrated, and the residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 40 g, 0-100% CH3CN / 10 mM NH4HCO3 in water) to give isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-(((decyloxy)carbonyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (139 mg, 0.180 mmol, 50.7% yield) as a white solid. LCMS (M+H) = 747.5; retention time (0.05% TFA) = 2.04 min. 1H NMR (400MHz, CDCl3) δ 8.02 (s, 1H), 7.28 (t, J = 8.4 Hz, 2H), 7.20 (d, J = 8.8 Hz, 2H), 7.12 (t, J = 6.8 Hz, 1H), 6.42 (t, J = 6.8 Hz, 1H), 6.09 (brs, 2H), 5.56 (dd, J = 7.2, 4.4 Hz, 1H), 5.00 (quint, J = 6.4 Hz, 1H), 4.46 (dd, J = 11.2, 6.4 Hz, 1H), 4.37 (dd, J = 11.2, 9.6 Hz, 1H), 4.19 (dt, J = 6.8, 2.0 Hz, 2H), 4.05-3.97 (m, 1H), 3.88 (dd, J = 11.2, 6.4 Hz, 1H), 2.85 (quint, J = 7.2 Hz, 1H), 2.76-2.70 (m, 1H), 2.68 (s, 1H), 1.70 (quint, J = 6.8 Hz, 2H), 1.36 (d, J = 7.2 Hz, 3H), 1.34-1.24 (m, 14H), 1.21 (d, J = 6.4 Hz, 6H), 0.88 (t, J = 6.8 Hz, 3H).

[0391] Example 50

[0392]

change

[0393] Project 1:4-ニトロフェニルペンタン-3-イルカーボネート A mixture of 4-nitrophenyl carbonochloridate (5 g, 24.81 mmol) and pentan-3-ol (2.187 g, 24.81 mmol) in DCM (100 mL) was treated with TEA (5.19 mL, 37.2 mmol), and the mixture was stirred at room temperature for 3 h. TLC showed that a new product had formed. Water (80 mL) was added, and the mixture was extracted with DCM (100 mL × 2). The combined organic phases were washed with brine, dried over Na2SO4, and concentrated to give 4-nitrophenylpentan-3-yl carbonate (6 g, 23.69 mmol, 96% yield). 1 H NMR (400MHz, CDCl3) δ 8.28 (dd, J = 7.2, 2.0 Hz, 2H), 7.39 (dd, J = 7.2, 2.0 Hz, 2H), 4.90-4.83 (m, 1H), 1.75-1.57 (m, 2H), 1.50-1.36 (m, 2H), 0.96 (t, J = 7.2 Hz, 6H).

[0394] Step 2: Isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((pentan-3-yloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate A mixture of isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (200 mg, 0.356 mmol) and DMAP (43.4 mg, 0.356 mmol) in DCM (6 mL) was treated with TEA (0.149 mL, 1.067 mmol) followed by 4-nitrophenylpentan-3-yl carbonate (360 mg, 1.422 mmol), and the mixture was stirred at 25° C. for 2 days. LCMS showed the reaction was complete. The reaction mixture was concentrated, and the residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 40 g, 0-100% CH3CN / 10 mM NH4HCO3 in water) to give isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((pentan-3-yloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (134 mg, 0.193 mmol, 54.4% yield) as a white solid. LCMS (M+H) = 677.2; retention time (0.05% TFA) = 1.73 min. 1H NMR (400MHz, CDCl3) δ 8.02 (s, 1H), 7.30-7.25 (m, 2H), 7.22-7.18 (m, 2H), 7.12 (t, J = 7.2 Hz, 1H), 6.43 (t, J = 6.8 Hz, 1H), 6.06 (brs, 2H), 5.56 (dd, J = 7.2, 4.0 Hz, 1H), 5.00 (quint, J = 6.4 Hz, 1H), 4.63 (quint, J = 6.4 Hz, 1H), 4.47 (dd, J = 11.2, 6.4 Hz, 1H), 4.39 (dd, J = 10.8, 6.0 Hz, 1H), 4.07-3.97 (m, 1H), 3.86 (dd, J = 11.2, 9.6 Hz, 1H), 2.83 (quint, J = 7.2 Hz, 1H), 2.71 (ddd, J = 14.0, 6.4, 4.0 Hz, 1H), 2.67 (s, 1H), 1.66 (quint, J = 6.8 Hz, 4H), 1.36 (d, J = 6.8 Hz, 3H), 1.21 (d, J = 6.4 Hz, 6H), 0.94 (dt, J = 7.6, 1.2 Hz, 6H).

[0395] Example 51

[0396]

change

[0397] Project 1: ヘプタン-4-イル(4-ニトロフェニル)カーボネート To a mixture of heptan-4-ol (3 g, 25.8 mmol) and DMAP (0.315 g, 2.58 mmol) in DCM (120 mL) was added TEA (5.38 mL, 38.7 mmol) dropwise at 0 °C. After stirring for 15 min, 4-nitrophenyl carbonochloridate (5.20 g, 25.8 mmol) was added. The resulting reaction mixture was stirred at 25 °C for 16 h. TLC indicated the reaction was complete. The reaction mixture was partitioned between DCM (120 mL) and water (50 mL). The organic layer was washed with brine (40 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified using silica gel column chromatography (hexane / ethyl acetate = 10 / 1) to give heptan-4-yl (4-nitrophenyl) carbonate (3.7 g, 13.15 mmol, 50.9% yield) as a colorless oil. 1 H NMR (400MHz, CDCl3) δ 8.28 (dd, J = 7.2, 2.0 Hz, 2H), 7.39 (dd, J = 7.2, 2.0 Hz, 2H), 4.90-4.83 (m, 1H), 1.75-1.57 (m, 4H), 1.50-1.36 (m, 4H), 0.96 (t, J = 7.2 Hz, 6H).

[0398] Step 2: Isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((heptan-4-yloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate A mixture of isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (200 mg, 0.356 mmol) and DMAP (43.4 mg, 0.356 mmol) in DCM (6 mL) was treated with TEA (0.149 mL, 1.067 mmol) followed by heptan-4-yl(4-nitrophenyl)carbonate (400 mg, 1.422 mmol), and the mixture was stirred at 25° C. for 2 days. LCMS showed the reaction was complete. The reaction mixture was concentrated, and the residue was dissolved in MeOH and purified using reverse-phase chromatography (SepaFlash® C18 column, Biotage; gradient: 0 to 100% ACN / 10 mM NH4HCO3 in water, flow rate: 40 mL / min, column: Boston 40 g) to afford isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((heptan-4-yloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (139 mg, 0.195 mmol, 54.8% yield) as a white solid. LCMS (M+H) = 705.3; retention time (0.05% TFA) = 1.84 min. 1H NMR (400MHz, CDCl3) δ 8.01 (s, 1H), 7.31-7.27 (m, 2H), 7.20 (d, J = 8.4 Hz, 2H), 7.12 (t, J = 7.2 Hz, 1H), 6.43 (t, J = 6.8 Hz, 1H), 5.92 (brs, 2H), 5.55 (dd, J = 6.8, 4.0 Hz, 1H), 5.00 (quint, J = 6.4 Hz, 1H), 4.81-4.74 (m, 1H), 4.47 (dd, J = 10.8, 6.0 Hz, 1H), 4.39 (dd, J = 11.2, 6.0 Hz, 1H), 4.06-3.97 (m, 1H), 3.80 (dd, J = 10.8, 9.6 Hz, 1H), 2.84 (quint, J = 7.2 Hz, 1H), 2.75-2.68 (m, 1H), 2.66 (s, 1H), 1.66-1.51 (m, 4H), 1.46-1.31 (m, 7H), 1.22 (d, J = 6.4 Hz, 6H), 0.93 (dt, J = 12.0, 7.2 Hz, 6H).

[0399] Example 52

[0400]

change

[0401] Project 1:4-ニトロフェニルノナン-5-イルカーボネート To a mixture of nonan-5-ol (3 g, 20.80 mmol) and DMAP (0.254 g, 2.080 mmol) in DCM (120 mL) was added TEA (3.16 g, 31.2 mmol) dropwise at 0 °C. After stirring for 15 min, 4-nitrophenyl carbonochloridate (4.19 g, 20.80 mmol) was added. The resulting reaction mixture was stirred at 25 °C for 16 h. TLC indicated the reaction was complete. The reaction mixture was partitioned between DCM (120 mL) and water (50 mL). The organic layer was washed with brine (40 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified using silica gel column chromatography (hexane / ethyl acetate = 10 / 1) to give 4-nitrophenylnonan-5-yl carbonate (4.08 g, 13.19 mmol, 63.4% yield) as a colorless oil. 1 H NMR (400MHz, CDCl3) δ 8.28 (dd, J = 7.2, 2.0 Hz, 2H), 7.39 (dd, J = 7.2, 2.0 Hz, 2H), 4.86-4.79 (m, 1H), 1.74-1.62 (m, 4H), 1.43-1.31 (m, 8H), 0.93 (t, J = 7.2 Hz, 6H).

[0402] Step 2: Isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((nonan-5-yloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate A mixture of isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (200 mg, 0.356 mmol) and DMAP (43.4 mg, 0.356 mmol) in DCM (6 mL) was treated with TEA (0.149 mL, 1.067 mmol) followed by 4-nitrophenylnonan-5-yl carbonate (440 mg, 1.422 mmol), and the mixture was stirred at 25° C. for 3 days. LCMS showed the reaction was complete. The reaction mixture was concentrated, and the residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 40 g, 0-100% CH3CN / 10 mM NH4HCO3 in water) to give isopropyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((nonan-5-yloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (102 mg, 0.137 mmol, 38.6% yield) as a white solid. LCMS (M+H) = 733.3; retention time (0.05% TFA) = 1.94 min. 1H NMR (400MHz, CDCl3) δ 8.02 (s, 1H), 7.31-7.27 (m, 2H), 7.20 (d, J = 8.4 Hz, 2H), 7.12 (t, J = 7.2 Hz, 1H), 6.43 (t, J = 6.8 Hz, 1H), 5.93 (brs, 2H), 5.55 (dd, J = 6.8, 4.0 Hz, 1H), 5.00 (quint, J = 6.4 Hz, 1H), 4.77-4.71 (m, 1H), 4.47 (dd, J = 11.2, 6.4 Hz, 1H), 4.39 (dd, J = 11.2, 6.0 Hz, 1H), 4.07-3.95 (m, 1H), 3.80 (dd, J = 10.8, 9.6 Hz, 1H), 2.81 (quint, J = 7.2 Hz, 1H), 2.71 (ddd, J = 14.0, 6.0, 4.0 Hz, 1H), 2.65 (s, 1H), 1.65-1.51 (m, 4H), 1.36 (d, J = 7.2 Hz, 3H), 1.35-1.25 (m, 8H), 1.22 (d, J = 6.0 Hz, 6H), 0.94-0.88 (m, 6H).

[0403] Example 53

[0404]

change

[0405] Process 1:2-エチルブチル((パーフルオロフェキシ)(フェノキシ)ホスホリル)-L-アラニネート To a stirred solution of 2-ethylbutyl L-alaninate hydrochloride (20 g, 95 mmol) in anhydrous dichloromethane (50 mL) was added TEA (10.0 g, 13.8 mL, 99.0 mmol) dropwise over 15 minutes at −70° C. To this mixture was added a solution of phenylphosphonic acid dichloride (18.41 g, 94 mmol) in anhydrous dichloromethane (80 mL) over 1 hour. The reaction mixture was stirred at this temperature for an additional 30 minutes, then warmed to 0° C. over 2 hours and stirred for an additional hour. To this mixture was added a solution of 2,3,4,5,6-pentafluorophenol (17.38 g, 94 mmol) and TEA (5.27 g, 7.26 mL, 52.1 mmol) in dichloromethane (60 mL) over 20 minutes. The crude mixture was stirred at 0° C. for 16 hours. LCMS indicated the reaction was complete. The mixture was concentrated under reduced pressure. The residue was triturated with TBME (800 mL) and triethylamine hydrochloride was removed by filtration. The cake was washed with TBME (2 × 50 mL) and the combined filtrates were concentrated under reduced pressure. The residue was triturated with 20% EtOAc in hexanes (250 mL) and the solid was collected by filtration to give 2-ethylbutyl ((perfluorophenoxy)(phenoxy)phosphoryl)-L-alaninate (7.8 g, 15.75 mmol, 16.51% yield) as a white solid. LCMS: (M+H) = 496.1; retention time (0.05% TFA) = 2.01 min.

[0406] Step 2: 2-Ethylbutyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate To a solution of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (0.25 g, 0.852 mmol) in THF (50 mL) and pyridine (2.500 mL) was added tert-butylmagnesium chloride (1.705 mL, 1.705 mmol) dropwise at −15° C. and stirred for 1 hour at −15° C. Next, a solution of 2-ethylbutyl ((S)-(perfluorophenoxy)(phenoxy)phosphoryl)-L-alaninate (0.549 g, 1.108 mmol) in THF (15 mL) was added, and the resulting reaction mixture was stirred at −15° C. for 2 hours. LCMS showed a 1:1 ratio of product to by-product. The reaction mixture was quenched with NH4Cl (4 mL) and partitioned between EtOAc (200 mL) and water (30 mL). The organic layer was washed with brine (20 mL), dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified using silica gel column chromatography (8% MeOH in DCM) to afford 2-ethylbutyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (130 mg, 0.213 mmol, 25.01% yield) as a white solid. LCMS: (M+H) = 605.3; retention time (0.05% TFA) = 1.61 min.

[0407] Step 3: (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((S)-(((S)-1-(2-ethylbutoxy)-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl nonanoate To a solution of nonanoic acid (51.0 mg, 0.323 mmol) in DCM (6 mL) were added N,N-dimethylpyridin-4-amine (30.3 mg, 0.248 mmol) and 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (95 mg, 0.496 mmol), and the resulting mixture was stirred at ice-water bath temperature for 0.5 h. (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((S)-(((S)-1-(2-ethylbutoxy)-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl nonanoate (100 mg, 0.130 mmol, 52.2% yield) was then added and the resulting mixture was stirred at 25° C. for 16 hours. LCMS showed the reaction was complete. The reaction mixture was concentrated, and the residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 40 g, 0-100% CH3CN / 10 mM NH4HCO3 in water) to afford (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((S)-(((S)-1-(2-ethylbutoxy)-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl nonanoate (100 mg, 0.130 mmol, 52.2% yield) as a white solid. LCMS (M+H) = 745.3; retention time (0.05% TFA) = 2.08 min. 1H NMR (400MHz, CDCl3) δ 8.07 (s, 1H), 7.31-7.27 (m, 2H), 7.23-7.19 (m, 2H), 7.13 (dt, J = 7.2, 0.8 Hz, 1H), 6.43 (t, J = 6.8 Hz, 1H), 6.22 (brs, 2H), 5.65 (dd, J = 6.8, 4.0 Hz, 1H), 4.43 (dd, J = 11.2, 6.4 Hz, 1H), 4.35 (dd, J = 11.2, 6.0 Hz, 1H), 4.11-4.04 (m, 2H), 3.99 (dd, J = 10.8, 5.6 Hz, 1H), 3.93 (dd, J = 11.2, 9.6 Hz, 1H), 2.71 (quint, J = 6.8 Hz, 1H), 2.64 (s, 1H), 2.64-2.59 (m, 1H), 2.40 (t, J = 7.6 Hz, 2H), 1.67 (quint, J = 7.2 Hz, 2H), 1.50 (quint, J = 6.0 Hz, 1H), 1.39 (d, J = 7.2 Hz, 3H), 1.37-1.25 (m, 14H), 0.92-0.83 (m, 9H).

[0408] Example 54

[0409] [ka]

[0410] (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((S)-(((S)-1-(2-ethylbutoxy)-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl decanoate To a solution of decanoic acid (55.6 mg, 0.323 mmol) in DCM (6 mL) were added N,N-dimethylpyridin-4-amine (30.3 mg, 0.248 mmol) and 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (95 mg, 0.496 mmol), and the resulting mixture was stirred at ice-water bath temperature for 0.5 h. Then, 2-ethylbutyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (150 mg, 0.248 mmol) was added, and the resulting mixture was stirred at 25 °C for 16 h. LCMS showed the reaction was complete. The reaction mixture was concentrated, and the residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 40 g, 0-100% CH3CN / 10 mM NH4HCO3 in water) to afford (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((S)-(((S)-1-(2-ethylbutoxy)-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl decanoate (108 mg, 0.137 mmol, 55.1% yield) as a white solid. LCMS (M+H) = 759.4; retention time (0.05% TFA) = 2.15 min. 1H NMR (400MHz, CDCl3) δ 8.13 (s, 1H), 7.31-7.27 (m, 2H), 7.23-7.19 (m, 2H), 7.13 (t, J = 7.2 Hz, 1H), 6.43 (t, J = 6.8 Hz, 1H), 5.95 (brs, 2H), 5.65 (dd, J = 6.8, 4.4 Hz, 1H), 4.43 (dd, J = 11.2, 6.4 Hz, 1H), 4.35 (dd, J = 10.8, 6.0 Hz, 1H), 4.11-4.04 (m, 2H), 3.99 (dd, J = 10.8, 6.0Hz, 1H), 3.77 (dd, J = 11.2, 9.6 Hz, 1H), 2.74-2.60 (m, 3H), 2.40 (t, J = 7.6 Hz, 2H), 1.67 (quint, J = 7.6 Hz, 2H), 1.51 (quint, J = 6.4 Hz, 1H), 1.39 (d, J = 6.8 Hz, 3H), 1.36-1.25 (m, 16H), 0.91-0.85 (m, 9H).

[0411] Example 55

[0412] [ka]

[0413] (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((S)-(((S)-1-(2-ethylbutoxy)-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl dodecanoate To a solution of dodecanoic acid (64.6 mg, 0.323 mmol) in DCM (6 mL) were added N,N-dimethylpyridin-4-amine (30.3 mg, 0.248 mmol) and 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (95 mg, 0.496 mmol), and the resulting mixture was stirred at ice-water bath temperature for 0.5 h. Then, 2-ethylbutyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (150 mg, 0.248 mmol) was added, and the resulting mixture was stirred at 25 °C for 16 h. LCMS indicated the reaction was complete. The reaction mixture was concentrated, and the residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 40 g, 0-100% CH3CN / 10 mM NH4HCO3 in water) to afford (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((S)-(((S)-1-(2-ethylbutoxy)-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl dodecanoate (122 mg, 0.153 mmol, 61.7% yield) as a white solid. LCMS (M+H) = 787.3; retention time (0.05% TFA) = 2.33 min. 1H NMR (400MHz, CDCl3) δ 8.04 (s, 1H), 7.31-7.27 (m, 2H), 7.22-7.19 (m, 2H), 7.13 (dt, J = 7.2, 0.8 Hz, 1H), 6.43 (t, J = 6.8 Hz, 1H), 5.87 (brs, 2H), 5.65 (dd, J = 7.2, 4.4 Hz, 1H), 4.43 (dd, J = 11.2, 6.0 Hz, 1H), 4.35 (dd, J = 10.8, 6.0 Hz, 1H), 4.11-4.04 (m, 2H), 3.99 (dd, J = 10.8, 6.0 Hz, 1H), 3.78 (dd, J = 11.2, 9.2 Hz, 1H), 2.73 (quint, J = 7.2 Hz, 1H), 2.66-2.60 (m, 2H), 2.40 (t, J = 7.6 Hz, 2H), 1.70-1.65 (m, 2H), 1.50 (quint, J = 6.4 Hz, 1H), 1.39 (d, J = 7.2 Hz, 3H), 1.35-1.25 (m, 20H), 0.91-0.85 (m, 9H).

[0414] Example 56

[0415] [ka]

[0416] (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((S)-(((S)-1-(2-ethylbutoxy)-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl tetradecanoate To a solution of tetradecanoic acid (73.7 mg, 0.323 mmol) in DCM (6 mL) were added N,N-dimethylpyridin-4-amine (30.3 mg, 0.248 mmol) and 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (95 mg, 0.496 mmol), and the resulting mixture was stirred at ice-water bath temperature for 0.5 h. Then, 2-ethylbutyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (150 mg, 0.248 mmol) was added, and the resulting mixture was stirred at 25 °C for 16 h. LCMS showed the reaction was complete. The reaction mixture was concentrated, and the residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 40 g, 0-100% CH3CN / 10 mM NH4HCO3 in water) to afford (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((S)-(((S)-1-(2-ethylbutoxy)-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl tetradecanoate (117 mg, 0.137 mmol, 55.1% yield) as a white solid. LCMS (M+H) = 815.4; retention time (0.05% TFA) = 2.55 min. 1H NMR (400MHz, CDCl3) δ 8.04 (s, 1H), 7.31-7.27 (m, 2H), 7.22-7.19 (m, 2H), 7.13 (t, J = 7.2 Hz, 1H), 6.43 (t, J = 6.8 Hz, 1H), 5.91 (brs, 2H), 5.65 (dd, J = 7.2, 4.4 Hz, 1H), 4.43 (dd, J = 11.2, 6.4 Hz, 1H), 4.35 (dd, J = 11.2, 6.0 Hz, 1H), 4.11-4.04 (m, 2H), 3.99 (dd, J = 10.8, 6.0 Hz, 1H), 3.79 (dd, J = 11.2, 9.6 Hz, 1H), 2.72 (quint, J = 6.8 Hz, 1H), 2.66-2.59 (m, 2H), 2.40 (t, J = 7.6 Hz, 2H), 1.70-1.67 (m, 2H), 1.50 (quint, J = 6.4 Hz, 1H), 1.39 (d, J = 6.8 Hz, 3H), 1.34-1.25 (m, 24H), 0.91-0.85 (m, 9H).

[0417] Example 57

[0418] [ka]

[0419] (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((S)-(((S)-1-(2-ethylbutoxy)-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl palmitate To a solution of palmitic acid (83 mg, 0.323 mmol) in DCM (6 mL) were added N,N-dimethylpyridin-4-amine (30.3 mg, 0.248 mmol) and 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (95 mg, 0.496 mmol), and the resulting mixture was stirred at ice-water bath temperature for 0.5 h. Then, 2-ethylbutyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (150 mg, 0.248 mmol) was added, and the resulting mixture was stirred at 20 °C for 16 h. LCMS showed the reaction was complete. The reaction mixture was concentrated, and the residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 40 g, 0-100% CH3CN / 10 mM NH4HCO3 in water) to afford (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((S)-(((S)-1-(2-ethylbutoxy)-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl palmitate (116 mg, 0.137 mmol, 55.1% yield) as a white solid. LCMS (M+H) = 843.1; retention time (0.1% TFA) = 3.30 min. 1H NMR (400MHz, CDCl3) δ 8.05 (s, 1H), 7.31-7.27 (m, 2H), 7.23-7.18 (m, 2H), 7.13 (t, J = 7.2 Hz, 1H), 6.43 (t, J = 6.8 Hz, 1H), 6.00 (brs, 2H), 5.65 (dd, J = 6.8, 4.4 Hz, 1H), 4.43 (dd, J = 11.2, 6.4 Hz, 1H), 4.35 (dd, J = 10.8, 6.0 Hz, 1H), 4.11-4.04 (m, 2H), 3.99 (dd, J = 10.8, 6.0 Hz, 1H), 3.84 (dd, J = 10.8, 9.2 Hz, 1H), 2.72 (quint, J = 6.8 Hz, 1H), 2.66-2.59 (m, 2H), 2.40 (t, J = 7.6 Hz, 2H), 1.70-1.62 (m, 2H), 1.50 (quint, J = 6.4 Hz, 1H), 1.39 (d, J = 7.2 Hz, 3H), 1.34-1.23 (m, 28H), 0.90-0.84 (m, 9H).

[0420] Example 58

[0421] [ka]

[0422] (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((S)-(((S)-1-(2-ethylbutoxy)-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl stearate To a solution of stearic acid (92 mg, 0.323 mmol) in DCM (6 mL) were added N,N-dimethylpyridin-4-amine (30.3 mg, 0.248 mmol) and 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (95 mg, 0.496 mmol), and the resulting mixture was stirred at ice-water bath temperature for 0.5 h. Then, 2-ethylbutyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (150 mg, 0.248 mmol) was added, and the resulting mixture was stirred at 20 °C for 16 h. LCMS showed the reaction was complete. The reaction mixture was concentrated, and the residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 40 g, 0-100% CH3CN / 10 mM NH4HCO3 in water) to afford (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((S)-(((S)-1-(2-ethylbutoxy)-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl stearate (151 mg, 0.173 mmol, 69.7% yield) as a white solid. LCMS (M+H) = 871.3; retention time (0.1% TFA) = 3.90 min. 1H NMR (400MHz, CDCl3) δ 8.04 (s, 1H), 7.31-7.27 (m, 2H), 7.23-7.19 (m, 2H), 7.13 (t, J = 6.8 Hz, 1H), 6.43 (t, J = 6.8 Hz, 1H), 5.92 (brs, 2H), 5.65 (dd, J = 7.2, 4.4 Hz, 1H), 4.43 (dd, J = 11.2, 6.0 Hz, 1H), 4.35 (dd, J = 10.8, 6.0 Hz, 1H), 4.11-4.04 (m, 2H), 3.99 (dd, J = 10.8, 6.0Hz, 1H), 3.80 (dd, J = 11.2, 9.6 Hz, 1H), 2.72 (quint, J = 6.8 Hz, 1H), 2.66-2.59 (m, 2H), 2.40 (t, J = 7.6 Hz, 2H), 1.66-1.62 (m, 2H), 1.50 (quint, J = 6.4 Hz, 1H), 1.39 (d, J = 7.2 Hz, 3H), 1.35-1.20 (m, 32H), 0.90-0.85 (m, 9H).

[0423] Example 59

[0424] [ka]

[0425] (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((S)-(((S)-1-(2-ethylbutoxy)-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl 2-propylpentanoate To a solution of 2-propylpentanoic acid (42.9 mg, 0.298 mmol) in DCM (6 mL) were added N,N-dimethylpyridin-4-amine (30.3 mg, 0.248 mmol) and 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (95 mg, 0.496 mmol), and the resulting mixture was stirred at ice-water bath temperature for 0.5 h. Then, 2-ethylbutyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (150 mg, 0.248 mmol) was added, and the resulting mixture was stirred at 25 °C for 16 h. LCMS indicated the reaction was complete. The reaction mixture was concentrated, and the residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 40 g, 0-100% CH3CN / 10 mM NH4HCO3 in water) to afford (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-((((S)-(((S)-1-(2-ethylbutoxy)-1-oxopropan-2-yl)amino)(phenoxy)phosphoryl)oxy)methyl)-2-ethynyltetrahydrofuran-3-yl 2-propylpentanoate (105 mg, 0.137 mmol, 55.3% yield) as a white solid. LCMS (M+H) = 731.2; retention time (0.05% TFA) = 1.99 min. 1H NMR (400MHz, CDCl3) δ 8.03 (s, 1H), 7.31-7.27 (m, 2H), 7.22-7.18 (m, 2H), 7.13 (dt, J = 7.2, 0.8 Hz, 1H), 6.42 (t, J = 6.8 Hz, 1H), 5.88 (brs, 2H), 5.66 (dd, J = 7.2, 5.2 Hz, 1H), 4.44 (dd, J = 11.2, 6.0 Hz, 1H), 4.33 (dd, J = 11.2, 6.0 Hz, 1H), 4.11-4.05 (m, 2H), 3.99 (dd, J = 10.8, 6.0 Hz, 1H), 3.81 (dd, J = 11.2, 9.6 Hz, 1H), 2.79 (quint, J = 6.8 Hz, 1H), 2.68-2.61 (m, 2H), 2.51-2.43 (m, 1H), 1.65-1.60 (m, 2H), 1.56-1.45 (m, 3H), 1.39 (d, J = 6.8 Hz, 3H), 1.36-1.30 (m, 8H), 0.91 (dt, J = 7.2, 1.2 Hz, 6H), 0.87 (t, J = 7.6, Hz, 6H).

[0426] Example 60

[0427] [ka]

[0428] 2-Ethylbutyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((hexyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate A mixture of 2-ethylbutyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (150 mg, 0.248 mmol) in DCM (10 mL) was treated with TEA (0.104 mL, 0.744 mmol), DMAP (30.3 mg, 0.248 mmol), and then hexyl(4-nitrophenyl)carbonate (265 mg, 0.992 mmol), and the resulting mixture was stirred at room temperature for 2 days. TLC showed the reaction was complete. The reaction mixture was concentrated in vacuo, and the residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 40 g, 0-100% CH3CN / 10 mM NH4HCO3 in water) to afford 2-ethylbutyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((hexyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (100 mg, 0.136 mmol, 55.0% yield) as a white solid. LCMS (M+H) = 733.7; retention time (0.1% TFA) = 1.94 min. 1H NMR (400MHz, CDCl3) δ 7.94 (s, 1H), 7.26 - 7.20 (m, 2H), 7.15 - 7.00 (m, 3H), 6.35 (t, J = 6.7 Hz, 1H), 6.11 - 5.84 (m, 2H), 5.50 (dd, J = 7.0, 4.4 Hz, 1H), 4.35 (ddd, J = 33.6, 11.1, 6.1 Hz, 2H), 4.12 (td, J = 6.7, 2.4 Hz, 2H), 4.04 - 3.90 (m, 2H), 3.87 - 3.73 (m, 1H), 2.77 (dd, J = 14.0, 7.0 Hz, 1H), 2.70 - 2.56 (m, 2H), 1.75 - 1.54 (m, 3H), 1.42 (dd, J = 12.5, 6.2 Hz, 1H), 1.32 - 1.18 (m, 10H), 0.81 (dt, J = 10.6, 7.1 Hz, 9H).

[0429] Example 61

[0430] [ka]

[0431] 2-Ethylbutyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((heptyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate A mixture of 2-ethylbutyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (150 mg, 0.248 mmol) in DCM (10 mL) was treated with TEA (0.104 mL, 0.744 mmol), DMAP (30.3 mg, 0.248 mmol), and then heptyl (4-nitrophenyl) carbonate (279 mg, 0.992 mmol), and the mixture was stirred at room temperature for 2 days. LCMS showed the reaction was complete. Water (20 mL) was added, and the mixture was then extracted with DCM (20 mL × 3). The combined organic phase was washed with brine (80 mL), dried over NaSO, concentrated in vacuo, and the residue was purified using silica gel column chromatography (MeOH / DCM = 0:100 to 1:20) to give 2-ethylbutyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((heptyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (106 mg, 0.141 mmol, 56.6% yield) as a white solid. LCMS (M+H) = 747.7; retention time (0.1% TFA) = 1.99 min. 1H NMR (400MHz, CDCl3) δ 7.94 (s, 1H), 7.26 - 7.20 (m, 2H), 7.15 - 7.00 (m, 3H), 6.35 (t, J = 6.7 Hz, 1H), 6.11 - 5.84 (m, 2H), 5.50 (dd, J = 7.0, 4.4 Hz, 1H), 4.35 (ddd, J = 33.6, 11.1, 6.1 Hz, 2H), 4.12 (td, J = 6.7, 2.4 Hz, 2H), 4.04 - 3.90 (m, 2H), 3.87 - 3.73 (m, 1H), 2.77 (dd, J = 14.0, 7.0 Hz, 1H), 2.70 - 2.56 (m, 2H), 1.75 - 1.54 (m, 3H), 1.42 (dd, J = 12.5, 6.2 Hz, 1H), 1.32 - 1.18 (m, 12H), 0.81 (dt, J = 10.6, 7.1 Hz, 9H).

[0432] Example 62

[0433] [ka]

[0434] 2-Ethylbutyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((octyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate A mixture of 2-ethylbutyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (200 mg, 0.331 mmol) in DCM (10 mL) was treated with TEA (0.138 mL, 0.992 mmol), DMAP (40.4 mg, 0.331 mmol), and then 4-nitrophenyloctyl carbonate (391 mg, 1.323 mmol), and the mixture was stirred at room temperature for 2 days. LCMS showed the reaction was complete. The reaction mixture was concentrated in vacuo, and the residue was purified using reverse-phase chromatography (SepaFlash® C18 column, BOSTON, 40 g, 0-100% CH3CN / 10 mM NH4HCO3 in water) to give 2-ethylbutyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((octyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (180 mg, 0.235 mmol, 71.2% yield) as a white solid. LCMS (M+H) = 761.7; retention time (0.1% TFA) = 2.04 min. 1H NMR (400MHz, CDCl3) δ 7.94 (s, 1H), 7.26 - 7.20 (m, 2H), 7.15 - 7.00 (m, 3H), 6.35 (t, J = 6.7 Hz, 1H), 6.11 - 5.84 (m, 2H), 5.50 (dd, J = 7.0, 4.4 Hz, 1H), 4.35 (ddd, J = 33.6, 11.1, 6.1 Hz, 2H), 4.12 (td, J = 6.7, 2.4 Hz, 2H), 4.04 - 3.90 (m, 2H), 3.87 - 3.73 (m, 1H), 2.77 (dd, J = 14.0, 7.0 Hz, 1H), 2.70 - 2.56 (m, 2H), 1.75 - 1.54 (m, 3H), 1.42 (dd, J = 12.5, 6.2 Hz, 1H), 1.32 - 1.18 (m, 14H), 0.81 (dt, J = 10.6, 7.1 Hz, 9H).

[0435] Example 63

[0436] [ka]

[0437] 2-Ethylbutyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((nonyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate A mixture of 2-ethylbutyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (200 mg, 0.331 mmol) and DMAP (40.4 mg, 0.331 mmol) in DCM (6 mL) was treated with TEA (0.138 mL, 0.992 mmol) followed by 4-nitrophenylnonyl carbonate (409 mg, 1.323 mmol), and the mixture was stirred at 20° C. for 3 days. LCMS showed the reaction was complete. The reaction mixture was concentrated, and the residue was purified using flash column chromatography (40 g, 0-4% MeOH in DCM) to afford 2-ethylbutyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-(((nonyloxy)carbonyl)oxy)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (132 mg, 0.166 mmol, 50.0% yield) as a white solid. LCMS (M+H) = 775.3; retention time (0.05% TFA) = 2.11 min. 1H NMR (400MHz, CDCl3) δ 8.06 (s, 1H), 7.30-7.25 (m, 2H), 7.22-7.18 (m, 2H), 7.12 (t, J = 7.2, 0.8 Hz, 1H), 6.42 (t, J = 6.8 Hz, 1H), 6.22 (brs, 2H), 5.56 (dd, J = 6.8, 4.4 Hz, 1H), 4.46 (dd, J = 11.2, 6.4 Hz, 1H), 4.38 (dd, J = 11.2, 6.0 Hz, 1H), 4.19 (dt, J = 6.8, 2.0 Hz, 2H), 4.11-4.04 (m, 2H), 3.99 (dd, J = 11.2, 6.0 Hz, 1H), 3.94 (dd, J = 11.2, 9.6 Hz, 1H), 2.83 (quint, J = 7.2 Hz, 1H), 2.72 (ddd, J = 14.0, 6.4, 4.4 Hz, 1H), 2.68 (s, 1H), 1.70 (quint, J = 6.8 Hz, 2H), 1.50 (quint, J = 6.4 Hz, 1H), 1.39 (dd, J = 7.2 Hz, 3H), 1.36-1.25 (m, 16H), 0.91-0.84 (m, 9H).

[0438] Example 64

[0439] [ka]

[0440] 2-Ethylbutyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-(((decyloxy)carbonyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate A mixture of 2-ethylbutyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxytetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (200 mg, 0.331 mmol) and DMAP (40.4 mg, 0.331 mmol) in DCM (6 mL) was treated with TEA (0.138 mL, 0.992 mmol) followed by decyl (4-nitrophenyl)carbonate (428 mg, 1.323 mmol), and the mixture was stirred at 20° C. for 3 days. LCMS showed the reaction was complete. The reaction mixture was concentrated, and the residue was purified using flash column chromatography (40 g, 0-4% MeOH in DCM) to afford 2-ethylbutyl ((S)-(((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-3-(((decyloxy)carbonyl)oxy)-2-ethynyltetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate (109 mg, 0.134 mmol, 40.4% yield) as a white solid. LCMS (M+H) = 789.3; retention time (0.05% TFA) = 2.18 min. ...

Claims

1. Compounds of formula (I): 【Chemistry 1】 [In the formula, R １ (C １ -C ２５ ) alkyl, (C １ -C ２５ ) alkylene aryl, (C １ -C ２０ ) alkylene-CO ２ R ５ and aryl; R ２ (C １ -C １０ ) alkyl, (C １ -C １０ ) selected from the group consisting of alkylene, aryl and aryl; R ３ (C １ -C ２５ ) alkyl, (C １ -C ２５ ) selected from the group consisting of alkylene, aryl and aryl; R ４ (C １ -C ２５ ) alkyl, (C １ -C ２５ ) alkylene aryl, (C １ -C ２０ ) alkylene-CO ２ R ５ and aryl; X is a bond, NR ６ or O, R ５ (C １ -C ２５ ) alkyl, (C １ -C ２５ ) selected from the group consisting of alkylene, aryl and aryl; R ６ is H, (C １ -C ２５ ) alkyl, (C １ -C ２５ ) alkylene aryl, (C １ -C ２０ ) alkylene-CO ２ R ５ and aryl; R １ , R ２ , R ３ , R ４ , R ５ and R ６ Each of independently represents one or more of (C １ -C １４ ) alkyl, Cl, F, oxo or (C １ -C ６ ) may be substituted with alkoxy. or a pharma- ceutically acceptable salt thereof.

2. R １ But (C ６ -C １４ 2. The compound of claim 1, or a pharma- ceutically acceptable salt thereof, wherein: R is 1 or 2; and R is 3 or 4.

3. R １ C ６ The compound according to any one of claims 1 to 2, or a pharma- ceutically acceptable salt thereof, which is aryl.

4. R １ But (C １ -C ２０ ) alkylene-CO ２ R ５ 2. The compound of claim 1, wherein:

5. R １ But (C １ -C ２０ ) alkylene-CO ２ R ５ is selected from R ５ But (C １ -C ２５ 2. The compound of claim 1, or a pharma- ceutically acceptable salt thereof, wherein: R is 1 or 2; and R is 3 or 4.

6. R ２ But (C １ -C １０ ) alkylene-(C ６ -C １４ 6. The compound according to claim 1, or a pharma- ceutically acceptable salt thereof, wherein: R is 1 or 2; and R is 3 or 4.

7. R ２ C １ Alkylene C ６ Aryl, C ６ Aryl is one or more (C １ -C １４ 7. The compound according to claim 1, which is optionally substituted by alkyl, Cl or F, or a pharma- ceutically acceptable salt thereof.

8. R ２ But (C １ -C ６ 6. The compound according to claim 1, wherein R is 1 or 2; or a pharma- ceutically acceptable salt thereof.

9. R ２ But (C １ 9. The compound according to any one of claims 1 to 8, or a pharma- ceutically acceptable salt thereof, wherein: R is 1 or 2; and R is 3 or 4.

10. R ３ But (C １ -C ２５ 10. The compound according to any one of claims 1 to 9, or a pharma- ceutically acceptable salt thereof, wherein: R is 1 or 2; and R is 3 or 4.

11. R ３ But (C １ -C ６ 11. The compound according to any one of claims 1 to 10, or a pharma- ceutically acceptable salt thereof, wherein: R is 1 or 2; and R is 3 or 4.

12. R ３ But (C １ -C １０ 11. The compound according to any one of claims 1 to 10, or a pharma- ceutically acceptable salt thereof, wherein: R is 1 or 2; and R is 3 or 4.

13. R ３ But (C １５ -C ２５ 10. The compound according to any one of claims 1 to 9, or a pharma- ceutically acceptable salt thereof, wherein: R is 1 or 2; and R is 3 or 4.

14. R ４ But (C １ -C ２５ 14. The compound according to any one of claims 1 to 13, or a pharma- ceutically acceptable salt thereof, wherein: R is 1 or 2; and R is 3 or 4.

15. R ４ But (C １ -C １０ 14. The compound according to any one of claims 1 to 13, or a pharma- ceutically acceptable salt thereof, wherein: R is 1 or 2; and R is 3 or 4.

16. R ４ But (C １５ -C ２０ 14. The compound according to any one of claims 1 to 13, or a pharma- ceutically acceptable salt thereof, wherein: R is 1 or 2; and R is 3 or 4.

17. 17. The compound of any one of claims 1 to 16, or a pharma- ceutically acceptable salt thereof, wherein X is a bond.

18. 17. The compound according to any one of claims 1 to 16, wherein X is O, or a pharma- ceutically acceptable salt thereof.

19. X is NR ６ 17. The compound according to any one of claims 1 to 16, wherein:

20. R １ C ６ aryl, R ２ C １ Alkyl or C １ Alkylene C ６ aryl, R ３ But (C １ -C ２５ ) alkyl, R ４ But (C １ -C ２５ 2. The compound of claim 1, wherein X is 1 or 2;

21. R １ C ６ aryl, R ２ C １ Alkyl or C １ Alkylene C ６ aryl, R ３ But (C １ -C ２５ ) alkyl, R ４ But (C １ -C ２５ 2. The compound of claim 1, wherein X is 0 or a pharma- ceutically acceptable salt thereof.

22. R １ C ６ aryl, R ２ C １ Alkyl or C １ Alkylene C ６ aryl, R ３ But (C １ -C ２５ ) alkyl, R ４ But (C １ -C ２５ ) alkyl, and X is NR ６ 2. The compound of claim 1, wherein:

23. R １ But (C １ -C ２５ ) alkylene-CO ２ R ５ and R ２ C １ Alkyl or C １ Alkylene C ６ aryl, R ３ But (C １ -C ２５ ) alkyl, R ４ But (C １ -C ２５ 2. The compound of claim 1, wherein X is 1 or 2;

24. R １ But (C １ -C ２５ ) alkylene-CO ２ R ５ and R ２ C １ Alkyl or C １ Alkylene C ６ aryl, R ３ But (C １ -C ２５ ) alkyl, R ４ But (C １ -C ２５ 2. The compound of claim 1, wherein X is 0 or a pharma- ceutically acceptable salt thereof.

25. R １ But (C １ -C ２５ ) alkylene-CO ２ R ５ and R ２ C １ Alkyl or C １ Alkylene C ６ aryl, R ３ But (C １ -C ２５ ) alkyl, R ４ But (C １ -C ２５ ) alkyl, and X is NR ６ 2. The compound of claim 1, wherein:

26. The following compound: 【Table 1】 or a pharma- ceutically acceptable salt thereof.

27. A pharmaceutical composition comprising a compound according to any one of claims 1 to 26, or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable excipient.

28. 28. The pharmaceutical composition of claim 27, wherein the pharmaceutical composition is in a parenteral form.

29. 28. The pharmaceutical composition of claim 27, wherein the pharmaceutical composition is in tablet form.

30. 28. The pharmaceutical composition of claim 27, wherein the pharmaceutical composition is formulated as a long-acting parenteral injection.

31. A pharmaceutical composition according to any one of claims 27 to 30 for treating HIV infection in a patient.

32. The pharmaceutical composition of any one of claims 27 to 30, for preventing HIV infection in a patient at risk of developing HIV infection.

33. A combination pharmaceutical comprising a compound according to any one of claims 1 to 26 or a pharma- ceutically acceptable salt thereof and one or more agents effective against HIV.

34. The combination pharmaceutical of claim 33, wherein the compound or pharma- ceutically acceptable salt thereof according to any one of claims 1 to 26 and one or more agents effective against HIV are administered simultaneously or sequentially in any order.

35. The pharmaceutical combination of claim 33 or 34 for treating HIV infection in a patient.

36. The pharmaceutical combination of claim 33 or 34 for preventing HIV infection in a patient at risk of developing HIV infection.

37. 27. A compound according to any one of claims 1 to 26, or a pharma- ceutically acceptable salt thereof, for use in the treatment of HIV infection.

38. 27. A compound according to any one of claims 1 to 26, or a pharma- ceutically acceptable salt thereof, for use in the prophylaxis of HIV infection.

39. 27. Use of a compound according to any one of claims 1 to 26, or a pharma- ceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of HIV infection.

40. 27. Use of a compound according to any one of claims 1 to 26, or a pharma- ceutically acceptable salt thereof, in the manufacture of a medicament for the prevention of HIV infection.

41. 27. Use of a combination comprising a compound according to any one of claims 1 to 26, or a pharma- ceutically acceptable salt thereof, and one or more agents effective against HIV, in the manufacture of a medicament for the treatment of HIV infection.

42. 27. Use of a combination comprising a compound according to any one of claims 1 to 26, or a pharma- ceutically acceptable salt thereof, and one or more agents effective against HIV, in the manufacture of a medicament for the prevention of HIV infection.

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