Adenosine derivatives and pharmaceutical compositions containing the same

Abstract

Disclosed herein are adenosine derivative prodrugs that can have reverse transcriptase inhibitor activity in vivo. The disclosure is also directed to pharmaceutical compositions containing adenosine derivatives that can be used to treat HIV infection or RNA virus infection. The disclosure is directed to adenosine derivative prodrugs that can inhibit reverse transcriptase. The disclosure is also directed to pharmaceutical compositions containing adenosine derivative prodrugs that can be used to treat acquired immune deficiency syndrome (AIDS), HIV-1, HIV-2, multidrug-resistant HIV, or a combination thereof.

Description

Technical Field

[0001] Cross - reference to Related Applications This application claims priority under 35 U.S.C. § 119(e) to U.S. Provisional Application No. 62 / 879,414, filed Jul. 27, 2019, which is hereby incorporated by reference in its entirety.

[0002] The present disclosure is directed to adenosine derivative prodrugs that can inhibit reverse transcriptase. The present disclosure is also directed to pharmaceutical compositions comprising adenosine derivative prodrugs that can be used in the treatment of acquired immunodeficiency syndrome (AIDS), HIV - 1, HIV - 2, multi - drug resistant HIV, or combinations thereof.

Background Art

[0003] Retroviruses such as human immunodeficiency virus (HIV) are associated with immunosuppressive diseases known as acquired immunodeficiency syndrome (AIDS). Multiple strains of retroviruses such as human immunodeficiency virus type 1 (HIV - 1) and human immunodeficiency virus type 2 (HIV - 2) are known to be associated with the disease. HIV retrovirus - infected individuals may initially be asymptomatic, but then develop AIDS - related complex (ARC) and subsequently AIDS. Replication of HIV by host cells requires integration of the viral genome into the host cell's DNA. An important step in the process is the transcription of the viral RNA genome into DNA via an enzyme known as reverse transcriptase (RT).

[0004] Reverse transcriptase can typically have multiple enzymatic functions that can act as (1) an RNA-dependent DNA polymerase that transcribes a single-stranded DNA copy (the first DNA) of viral RNA, (2) a ribonuclease that destroys the original viral RNA and releases the DNA just produced from the original RNA, and (3) a DNA-dependent DNA polymerase that uses the first DNA strand as a template to produce a second complementary DNA strand. The two DNA strands then form double-stranded DNA, which is integrated into the genome of the host cell by the integrase enzyme.

[0005] Some compounds can inhibit reverse transcriptase (RT) activity. These compounds may be useful for the treatment of HIV infection in humans by inhibiting HIV replication in infected cells or individuals. Examples of compounds approved for use in the treatment of HIV infection and AIDS include nucleoside RT inhibitors (NNRTIs) such as 3'-azido-3'-deoxythymidine (AZT, also known as zidovudine (ZDV), azidothymidine (AZT)), 2',3'-dideoxyinosine (ddI), 2',3'-dideoxycytidine (ddC), d4T, 3TC, abacavir, emtricitabine, and tenofovir disoproxil fumarate, as well as nevirapine, delavirdine, efavirenz, rilpivirine, and dolutegravir (DHHS guidelines: https: / / aidsinfo.nih.gov / understanding-hiv-aids, Iyidogan & Anderson, Viruses, 6, 4095 - 4139, 2014, doi:10.3390 / v6104095; Hayakawa et al., Antiviral Chem & Chemotherapy, 15:169 - 187, 2004; Ohrul et al., J. Med. Chem. 43, 4516 - 4525, 2000; Pauwels, Antiviral Research, 71, 77 - 89, 2006). The adenosine derivative EFdA (4’-ethynyl-2-fluoro-2’-deoxyadenosine, also known as MK-8591 and islatravir) is a long-acting (LA) NRTI that has been demonstrated to have anti-HIV activity by inhibiting reverse transcriptase by preventing translocation (U.S. Patent Nos. 7,339,053, 7,625,877, 8,039,614; Singh et al., Pharmaceuticals, 12, 62, 2019, DOI: 10.3390 / ph12020062, each of which is incorporated herein by reference in its entirety). This compound has broad inhibitory activity and efficacy against HIV-1, HIV-2, as well as different subtypes and mutations, including multi-drug resistant (MDR) and wild-type (WT) strains, and reverse transcriptase inhibitor (RTI)-resistant viruses. Some modified EFdA and prodrugs are described in U.S. Patent Publication No. 2018 / 0002366, which is incorporated herein by reference in its entirety. A common problem arising from the treatment of HIV infections with anti-retroviral inhibitory compounds is viral resistance to the inhibitor. Such resistance is typically the result of mutations occurring within the reverse transcriptase segment of the pol gene. The continued use of anti-viral compounds, such as inhibitory compounds for preventing HIV infections, will inevitably lead to the emergence of new resistant strains of HIV. Thus, there is a continuing need for new RT inhibitors that are effective against HIV strains, including mutant HIV and multi-drug resistant HIV strains.

Prior Art Documents

Patent Documents

[0006]

Patent Document 1

Patent Document 2

Patent Document 3

Patent Document 4

Non-Patent Literature

[0007]

Non-Patent Literature 1

Non-Patent Literature 2

Non-Patent Literature 3

Non-Patent Literature 4

Non-Patent Literature 5

Summary of the Invention

Means for Solving the Problems

[0008] The present disclosure relates to adenosine derivatives and compositions thereof that can be used for the treatment of retroviral diseases such as HIV and AIDS.

[0009] In some embodiments, the present disclosure provides an adenosine derivative having formula (1) or a stereoisomer, pharmaceutically acceptable salt, tautomer, or solvate thereof,

Chemical Formula

Chemical formula

[0010] In some embodiments, the adenosine derivative is Formula (2-A):

Chemical formula

Chemical formula

[0011] In some embodiments of formula (1), the adenosine derivative is R 1 , R 1’、 R 2 , or contains a structure selected from formulas 9 - 24. [Chem.]

[0012] The present disclosure further relates to a pharmaceutical composition comprising one or more adenosine derivatives disclosed herein, their pharmaceutically acceptable salts, stereoisomers, or combinations thereof, and one or more pharmaceutically acceptable carriers.

[0013] The present disclosure also relates to a method for treating a disease (e.g., acquired immunodeficiency syndrome (AIDS) or human immunodeficiency virus (HIV)), the method comprising administering to a subject in need thereof a pharmaceutical composition comprising an effective dosage of one or more of the adenosine derivatives disclosed herein.

[0014] The present disclosure also relates to a method for preventing infection, the method comprising administering to a subject in need thereof a pharmaceutical composition comprising an effective dosage of one or more of the adenosine derivatives disclosed herein.

[0015] Incorporation by reference All publications, patents, and patent applications mentioned herein are incorporated herein by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. BRIEF DESCRIPTION OF THE DRAWINGS

[0016]

Figure 1A

Figure 1B

Figure 1C

Figure 1D

Figure 1E

Figure 1F

Figure 1G

Figure 1H

Figure 1I

Figure 2A

Figure 2B

Figure 2C

Figure 2D

Figure 2E

Figure 2F

Figure 2G

Figure 2H

Figure 2I

Figure 3

Figure 4

Figure 5

[0017] The following is a more detailed description of various concepts and their embodiments related to the methods and apparatuses according to the present disclosure. It should be understood that since the subject matter is not limited to any particular embodiment, the various aspects of the subject matter introduced above and discussed in more detail below can be implemented in any of a number of ways. Examples of specific implementations and applications are provided primarily for illustrative purposes.

[0018] As used herein, the term "alkyl" or "alkyl group" refers to a fully saturated, straight-chain or branched hydrocarbon chain having 1 to 12 carbon atoms and attached to the remainder of the molecule by a single bond. Alkyls containing any number of carbon atoms from 1 to 12 are included. Alkyls containing up to 12 carbon atoms are C1-C 12 alkyl, and alkyls containing up to 10 carbon atoms are C1-C 10 alkyl, and alkyls containing up to 6 carbon atoms are C1-C6 alkyl, and alkyls containing up to 5 carbon atoms are C1-C5 alkyl. C1-C5 alkyl includes C5 alkyl, C4 alkyl, C3 alkyl, C2 alkyl, and C1 alkyl (i.e., methyl). C1-C6 alkyl includes all of the moieties described above for C1-C5 alkyl, but also includes C6 alkyl. C1-C 10 alkyl includes all of the moieties described above for C1-C5 alkyl and C1-C6 alkyl, but also includes C7, C8, C9, and C 10 alkyl. Similarly, C1-C 12Alkyl includes all of the foregoing moieties, but also includes C 11 and C 12 alkyl. Non-limiting examples of C1-C 12 alkyl include methyl, ethyl, n-propyl, i-propyl, sec-propyl, n-butyl, i-butyl, sec-butyl, t-butyl, n-pentyl, t-amyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, n-undecyl, and n-dodecyl. Unless otherwise specified herein, an alkyl group may be optionally substituted.

[0019] As used herein, the term "alkylene" or "alkylene chain" refers to a fully saturated, straight-chain or branched-chain divalent hydrocarbon chain radical having from 1 to 12 carbon atoms. Non-limiting examples of C1-C 12 alkylene include methylene, ethylene, propylene, n-butylene, etc. The alkylene chain is bonded to the remainder of the molecule via a single bond and to a radical group (e.g., a group described herein) via a single bond. The points of attachment of the alkylene chain to the remainder of the molecule and to the radical group can be via one carbon or any two carbons within the chain. Unless otherwise specified herein, the alkylene chain may be optionally substituted.

[0020] As used herein, the term "alkenyl" or "alkenyl group" refers to a straight-chain or branched-chain aliphatic hydrocarbon radical containing at least one carbon-carbon double bond and having a specified number of carbon atoms in the range. For example, "C2-C10 alkenyl" (or "C2-C 10"(Alkenyl") refers to either a linear or branched alkenyl having 2 to 10 carbon atoms or an isomer thereof. In another embodiment, C2-C6 alkenyl can have 1-butenyl, 2-butenyl, 3-butenyl, isobutenyl, 1-propenyl, 2-propenyl, and ethenyl (or vinyl). The remainder of the alkylene chain molecule and the point of attachment to the radical group can be through one carbon or any two carbons within the chain. Unless otherwise specified herein, the alkylene chain can be optionally substituted.

[0021] As used herein, the term "alkenylene" or "alkenylene chain" refers to an unsaturated, linear or branched divalent hydrocarbon chain radical having one or more olefins and 2 to 12 carbon atoms. C2-C 12 Non-limiting examples of alkenylene include ethenylene, propenylene, n-butenylene, and the like. The alkenylene chain is attached to the remainder of the molecule through a single bond and to a radical group (e.g., a group described herein) through a single bond. The point of attachment of the alkenylene chain to the remainder of the molecule and to the radical group can be through one carbon or any two carbons within the chain. Unless otherwise specified herein, the alkenylene chain can be optionally substituted.

[0022] As used herein, the term "cycloalkyl" can include a fused or bridged ring system having from 3 to 20 carbon atoms (e.g., having from 3 to 10 carbon atoms), and refers to a stable non-aromatic monocyclic or polycyclic fully saturated hydrocarbon consisting only of carbon and hydrogen atoms, which is bonded to the remainder of the molecule by a single bond. Examples of monocyclic cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Examples of polycyclic cycloalkyl include adamantyl, norbornyl, decalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like. In some embodiments, "cycloalkyl" refers to any monocyclic ring of an alkane having the number of carbon atoms within the specified range. For example, "C3-C10 cycloalkyl" (or "C3-C 10 cycloalkyl") refers to a monocyclic ring of an alkane having from 3 to 10 carbon atoms such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl. Unless otherwise specified herein, cycloalkyl groups can be optionally substituted.

[0023] As used herein, the terms "heterocycloalkyl", "heterocyclic ring", or "heterocycle" refer to a saturated or partially saturated 3- to 20-membered ring consisting of 2 to 19 carbon atoms and 1 to 6 heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur, and attached to the remainder of the molecule by a single bond. Unless otherwise specified herein, a heterocycloalkyl can be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which can include fused or bridged ring systems, and the nitrogen, carbon, or sulfur atoms in the heterocycloalkyl can be optionally oxidized, for example, to form N-oxides, sulfoxides, or sulfones, and / or the nitrogen atoms can be optionally quaternized to form, for example, quaternary ammonium cations. Examples of such heterocycloalkyls include, but are not limited to, dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. In some embodiments, "3- to 10-membered heterocycloalkyl" refers to a cycloalkyl containing one or more heteroatoms selected from the group consisting of N, O, and S. In some embodiments, "heterocycloalkyl", "heterocyclic ring", or "heterocycle" refers to a 3- to 10-membered ring structure having, as members of the ring structure, carbon atoms and one or more heteroatoms selected from N, O, S, or combinations thereof. Unless otherwise specified herein, a heterocycloalkyl group can be optionally substituted and can include saturated and / or unsaturated rings.

[0024] As used herein, the term "halogen" (or "halo") refers to fluorine, chlorine, bromine, and iodine (alternatively referred to as fluoro (-F), chloro (-Cl), bromo (-Br), and iodo (-I)).

[0025] As used herein, the term "aryl" refers to a hydrocarbon ring system that contains hydrogen, from 6 to 18 carbon atoms, and at least one aromatic ring and is bonded to the remainder of the molecule by a single bond. For the purposes of the present disclosure, aryl can be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which can include fused or bridged ring systems. Examples of aryl include, but are not limited to, aryls derived from aceanthrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, chrysene, fluoranthene, fluorene, as-indacene, s-indacene, indane, indene, naphthalene, phenalene, phenanthrene, preiadene, pyrene, and triphenylene. In some embodiments, "aryl" refers to phenyl or one or more fused cyclic hydrocarbon ring systems in which at least one ring is aromatic. Unless otherwise specified herein, "aryl" can be optionally substituted.

[0026] As used herein, the term "heteroaryl" refers to a 5- to 20-membered ring system containing a hydrogen atom, 1 to 19 carbon atoms, 1 to 6 heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur, at least one aromatic ring, and being bonded to the remainder of the molecule by a single bond. For the purposes of the present disclosure, heteroaryl can be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which can include fused or bridged ring systems, and the nitrogen, carbon, or sulfur atoms in the heteroaryl can be optionally oxidized, for example, to form N-oxides, sulfoxides, or sulfones, and / or the nitrogen atoms can be optionally quaternized to form, for example, quaternary ammonium cations. Non-limiting examples of heteroaryl include pyridyl, pyrrolyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, thienyl, furanyl, imidazolyl, pyrazolyl, triazolyl (i.e., 1,2,3-triazolyl or 1,2,4-triazolyl), tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl (i.e., 1,2,3-, 1,2,4-, 1,2,5-(furazanyl), or 1,3,4-isomers), oxatriazolyl, thiazolyl, isothiazolyl, and thiadiazolyl. Suitable 9- and 10-membered bicyclic fused ring systems include, for example, benzofuranyl, indolyl, indazolyl, naphthyridinyl, isobenzofuranyl, benzopiperidinyl, benzyisoxazolyl, benzoxazolyl, chromenyl, quinolinyl, isoquinolinyl, cinnolinyl, quinazolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, isoindolyl, benzodioxolyl, benzopiperidinyl, benzyisoxazolyl, benzoxazolyl, chromanyl, isochromanyl, benzothienyl, benzofuranyl, imidazo[1,2-a]pyridinyl, benzotriazolyl, dihydroindolyl, dihydroisoindolyl, indazolylinyl, indolinyl, isoindolinyl, quinoxalinyl, quinazolinyl, 2,3-dihydrobenzofuranyl, and 2,3-dihydrobenzo-1,4-dioxinyl. Unless otherwise specified herein, heteroaryl groups can be optionally substituted.

[0027] Unless expressly stated to the contrary in a particular context, it is understood that any of the various cyclic rings and ring systems described herein may be attached at any ring atom (i.e., any carbon atom or any heteroatom) to the remainder of the compound, provided that the attachment is chemically acceptable.

[0028] As used herein, the term "substituted" means that at least one hydrogen atom has been replaced by a bond to a non-hydrogen atom such as, but not limited to, a halogen atom such as F, Cl, Br, and I, an oxygen atom in a group such as a hydroxyl group, an alkoxy group, and an ester group, a sulfur atom in a group such as a thiol group, a thioalkyl group, a sulfone group, a sulfonyl group, and a sulfoxide group, a nitrogen atom in a group such as an amine, an amide, an alkylamine, a dialkylamine, an arylamine, an alkylarylamine, an N-oxide, an imide, and an enamine, a silicon atom in a group such as a trialkylsilyl group, a dialkylarylsilyl group, an alkyldiarylsilyl group, and a triarylsilyl group, and other heteroatoms in various other groups, of the groups described herein (e.g., alkyl, alkenyl, alkynyl, alkoxy, aryl, aralkyl, carbocyclic, cycloalkyl, cycloalkenyl, cycloalkynyl, haloalkyl, heterocyclyl, and / or heteroaryl). "Substitution" also means that one or more hydrogen atoms have been replaced by a higher order bond (e.g., a double bond or a triple bond) to a heteroatom such as oxygen in an oxo, carbonyl, carboxyl, and ester group, and nitrogen in a group such as an imine, an oxime, a hydrazone, and a nitrile. For example, "substituted" means that one or more hydrogen atoms are -NR g R h 、 -NR g C(=O)R h 、 -NR g C(=O)NR g R h 、 -NR g C(=O)OR h 、 -NR g SO2R h 、 -OC(=O)NRg R h , -OR g ,-SR g , -SOR g , -SO2R g , -OSO2R g , -SO2OR g , =NSO2R g , and -SO2NR g R h "Substituted" also refers to any of the above groups in which one or more hydrogen atoms have been replaced with -C(=O)R. g , -C(=O)OR g , -C(=O)NR g R h , -CH2SO2R g , -CH2SO2NR g R h In the above, R g and R h are the same or different and are independently hydrogen, alkyl, alkenyl, alkynyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkenyl, cycloalkylalkyl, haloalkyl, haloalkenyl, haloalkynyl, heterocyclyl, N-heterocyclyl, heterocyclylalkyl, heteroaryl, N-heteroaryl, and / or heteroarylalkyl. "Substituted" further refers to any of the above groups in which one or more hydrogen atoms are replaced by a bond to an amino, cyano, hydroxyl, imino, nitro, oxo, thioxo, halo, alkyl, alkenyl, alkynyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkenyl, cycloalkynyl, cycloalkylalkyl, haloalkyl, haloalkenyl, haloalkynyl, heterocyclyl, N-heterocyclyl, heterocyclylalkyl, heteroaryl, N-heteroaryl, and / or heteroarylalkyl group. In addition, each of the foregoing substituents may be optionally substituted with one or more of the above substituents.

[0029] As used herein, the term "isomer" refers to structural isomers such as groups or atoms arranged at different positions in a molecule, stereoisomers such as chiral isomers, enantiomers, diastereomers and cis / trans isomers, amino isomers, imino isomers, or tautomers such as combinations thereof. By way of non-limiting example, the adenosine derivatives of the present disclosure may have amino isomers, imino isomers, or combinations thereof. By way of another non-limiting example, where an -OH substituent is tolerated on a heteroaromatic ring and keto-enol tautomerism is possible, it is understood that the substituent may actually be present in whole or in part in the oxo (=O) form. Mixtures of isomers may also be suitable. A mixture of isomers may contain each isomer in all ratios. Salts of isomers may also be suitable. The adenosine derivatives of the present disclosure may contain one or more of its isomers, one or more salts thereof, one or more solvates containing its hydrate, its solvated salts, or mixtures thereof. Absolute stereochemistry or isomeric constitution may be determined by X-ray crystallography, by vibrational circular dichroism (VCD) spectroscopy, or by a combination thereof.

[0030] Adenosine derivatives can be identified by a name based on the nomenclature recommended by the International Union of Pure and Applied Chemistry (IUPAC), or based on the name of a nucleoside (nucleoside-based nomenclature). Adenosine derivatives can also be identified by a chemical structure diagram. Unless explicitly stated to the contrary in a particular context, the name and the structure may be used interchangeably.

[0031] Any atom in the compounds disclosed herein may exhibit their natural isotopic abundances, or one or more of the atoms may be artificially enriched in a particular isotope having the same atomic number but a different atomic mass or mass number than the atomic mass or mass number predominantly found in nature. The present disclosure is meant to include all suitable isotopic variations of the compounds disclosed herein.

[0032] The compound can be administered in the form of a pharmaceutically acceptable salt or solvate. The term "pharmaceutically acceptable salt" refers to salts or solvates that are not biologically or otherwise undesirable (e.g., not toxic or harmful to the recipient or the subject thereof). Mixtures of the compounds disclosed herein with one or more of their salts or solvates are also contemplated herein. Exemplary examples of pharmaceutically acceptable salts include, but are not limited to, sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caproate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, butyne-1,4-dioate, hexyne-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, sulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, y-hydroxybutyrate, glycolate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, and mandelate.

[0033] Furthermore, the compounds disclosed herein can exist in amorphous form and / or one or more crystalline forms, or combinations thereof.

[0034] The term "RNA virus infection" refers to diseases caused by RNA viruses such as colds, influenza, SARS, COVID-19, hepatitis C, hepatitis E, West Nile fever, Ebola virus disease, rabies, polio, and measles.

[0035] The term "HIV infection" refers to a disease caused by human immunodeficiency virus (HIV) such as HIV-1 and HIV-2. In some cases, HIV infection can be caused by wild-type HIV-1, NRTI-resistant HIV-1, HIV-2, HIV with the M184V mutation, HIV with K65R, or multi-drug resistant HIV. The term "AIDS" refers to acquired immunodeficiency syndrome caused by HIV infection and the progressive form of the disease.

[0036] The term "prodrug" refers to a compound that can be converted into a biologically active compound described herein under physiological conditions or by solvolysis. Thus, the term "prodrug" refers to a precursor of a pharmaceutically acceptable biologically active compound. A prodrug may be a biologically inactive or substantially inactive compound, which can be metabolized in the body, i.e., in vivo, to produce a drug having the desired activity. The term "substantially inactive" means that the prodrug may have an activity ratio of about 1% to about 10% of the corresponding drug or after metabolism in vivo, based on the weight of the prodrug. In some embodiments, the term "substantially inactive" means that the prodrug may have an activity ratio of less than about 5% of the corresponding drug or after metabolism in vivo, based on the weight of the prodrug. The doses of the prodrug and its biologically active compound are considered to be dose-equivalent when they are in the same molar amount.

[0037] The terms "anti-HIV agent", "antiviral agent", or grammatical variants thereof refer to compounds, mixtures of one or more compounds, formulations, chemical agents, or biological agents such as antibodies, proteins, peptides, nucleotides, and other biological compounds that can be directly or indirectly effective in the inhibition of HIV, the treatment or prevention of HIV infection, and / or AIDS and / or diseases or conditions resulting from or associated with them, RNA virus infections, or combinations thereof. Anti-HIV agents can include anti-HIV antiviral agents, immunomodulators, anti-infective agents, vaccines, or combinations thereof that are useful for the treatment of HIV infection or AIDS. Examples of antiviral agents for the treatment of HIV infection or AIDS include, under their respective trademarks or registered trademarks with their respective owners, abacavir (ABC, Ziagen®), abacavir + lamivudine (Epzicom®), abacavir + lamivudine + zidovudine (Trizivir®), amprenavir (Agenerase®), atazanavir (Reyataz®), AZT (zidovudine, azidothymidine or Retrovir®), capravirine, darunavir (Prezista®), ddC (zalcitabine, dideoxycytidine or Hivid®), ddI (didanosine, dideoxyinosine or Videx®), ddI (enteric-coated, VidexEC (registered trademark)), Delavirdine (DLV or Rescriptor (registered trademark)), Dolutegravir (Tivicay (registered trademark)), Dravirine (MK-1439), Efavirenz (EFV, Sustiva (registered trademark), Stocrin (registered trademark)), Efavirenz + Emtricitabine + Tenofovir DF (Atripla (registered trademark)), EFdA (4'-Ethynyl-2-fluoro-2'-deoxyadenosine), Elvitegravir, Cabotegravir, Dolutegravir, Bictegravir, Emtricitabine (FTC, Emtriva (registered trademark)), Emtricitabine + Tenofovir DF (Truvada (registered trademark)), Embiline (Coactinon (registered trademark)), Enfuvirtide (Fuzeon (registered trademark)), Enteric-coated Didanosine (VidexEC (registered trademark), etravirine (TMC-125), fosamprenavir calcium (Lexiva (registered trademark)), indinavir (Crixivan (registered trademark)), lamivudine (3TC, Epivir (registered trademark)), lamivudine + zidovudine (Combivir (registered trademark)), lopinavir, lopinavir + ritonavir (Kaletra (registered trademark)), maraviroc (Selzentry (registered trademark)), nelfinavir (Viracept (registered trademark)), nevirapine (NVP, Viramune (registered trademark)), PPL-100 (also known as PL-462 (Ambrilia)), raltegravir (MK-0518 or Isentress (trademark)), rilpivirine (Edurant (registered trademark)), ritonavir (Norvir (registered trademark)), saquinavir (Invirase (registered trademark), or Fortovase (registered trademark)), stavudine (d4T, didehydrodeoxythymidine or Zerit (registered trademark)), tenofovir DF (DF = disoproxil fumarate, TDF, Viread (registered trademark)), tenofovir (hexadecyloxypropyl (CMX-157)), tenofovir alafenamide fumarate (GS-7340), tipranavir (Aptivus (registered trademark)) and vicriviroc, and the like, but not limited thereto. Some of the above anti-HIV agents can be used in salt form, for example, abacavir sulfate, delavirdine mesylate, indinavir sulfate, atazanavir sulfate, nelfinavir mesylate, saquinavir mesylate, or other salts. The anti-HIV agent can have one or more activities such as entry inhibitor (EI), fusion inhibitor (FI), integrase inhibitor (InI), protease inhibitor (PI), nucleoside reverse transcriptase inhibitor (nRTI or NRTI), or non-nucleoside reverse transcriptase inhibitor (nnRTI or NNRTI). The anti-HIV agent can comprise two or more agents disclosed herein. The adenosine derivative of the present disclosure can be an anti-HIV agent together with or in combination with other anti-HIV agent(s).

[0038] Unless explicitly stated to the contrary, all ranges recited herein include their endpoints. For example, a heteroaryl ring described as being within the range of "1 to 4 heteroatoms" means that the ring can contain 1, 2, 3, or 4 heteroatoms. It should also be understood that any range recited herein includes all sub-ranges within that range. Thus, for example, a heterocyclic ring described as containing "1 to 4 heteroatoms" includes, as aspects thereof, heterocyclic rings containing 2 to 4 heteroatoms, 3 to 4 heteroatoms, 1 to 3 heteroatoms, 2 to 3 heteroatoms, 1 to 2 heteroatoms, 1 heteroatom, 2 heteroatoms, 3 heteroatoms, or 4 heteroatoms. In other examples, C1-C10 alkyl means alkyl containing 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10 carbon atoms, including all sub-ranges. Thus, C1-C10 alkyl can be straight-chain or branched methyl, ethyl, C4 alkyl, C5 alkyl, C6 alkyl, C7 alkyl, C8 alkyl, C9 alkyl, and C10 alkyl. Divalent C1-C10 alkyl can be straight-chain or branched -CH2-, -C2H4-, -C3H6-, -C4H8-, -C5H 10 -, -C6H 12 -, -C7H 17 -, -C8H 18 -, -C9H 18 - or -C 10 H 20 -. Similarly, C2-C10 alkenyl means straight-chain or branched alkenyl containing 2, 3, 4, 5, 6, 7, 8, 9, and 10 carbon atoms, including all sub-ranges. Straight-chain or branched alkenyl can be preferred. C3-C10 cycloalkyl means cycloalkyl containing 3, 4, 5, 6, 7, 8, 9, and 10 carbon atoms, straight-chain or branched.

[0039] Unless otherwise stated, open terms such as, for example, "contain", "containing", "include", "including", etc. mean "comprising".

[0040] The singular forms "a", "an", and "the" are used herein to include a plurality of referents unless otherwise clearly indicated in the context. Thus, unless the contrary is indicated, the numerical parameters set forth in this application are approximations that can vary depending upon the desired properties sought to be obtained by the present disclosure.

[0041] The term "about" and its grammatical equivalents as related to a reference numerical value used herein can include values that are within plus or minus 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% of that value, such as values within plus or minus 10% of that value. For example, an amount of "about 10" can include amounts from 9 to 11.

[0042] The pharmaceutical composition can be suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal, or intracutaneous administration (e.g., by injection or infusion). Depending on the route of administration, the active ingredient can be coated with a material to protect it from the action of acids and other natural conditions that can inactivate the active ingredient. As used herein, the phrase "parenteral administration" generally means a mode of administration other than enteral and topical administration by injection and includes, but is not limited to, intravenous, intramuscular, arterial, intrathecal, intracapsular, intraorbital, intracardiac, intracutaneous, intraperitoneal, transtracheal, subcutaneous, subepidermal, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injection and infusion. Alternatively, the pharmaceutical composition can be administered via non-parenteral routes such as local, epidermal, or mucosal administration routes such as intranasal, oral, intravaginal, rectal, sublingual, or topical. The pharmaceutical composition can be in the form of a sterile aqueous solution or dispersion. The pharmaceutical composition can also be formulated in microemulsions, liposomes, or other ordered structures suitable for high drug concentrations.

[0043] In some embodiments, the present disclosure provides an adenosine derivative having formula (1) or a pharmaceutically acceptable salt, tautomer, or solvate thereof,

Chemical Formula

Chemical formula

[0044] The adenosine derivatives of the present disclosure may not contain a monophosphate group, a diphosphate group, a triphosphate group, or a combination thereof. In some embodiments, R of the adenosine derivative of the present disclosure 1 , R 1’ or R 2 groups do not contain a monophosphate group, a diphosphate group, a triphosphate group, or a combination thereof. Non-limiting examples of adenosine derivatives having a halogen atom are shown in formulas (1) to (8) and (4-B) (FIGS. 1A to 1I).

[0045] In some embodiments, the C1-C10 alkyl and C2-C10 alkenyl in formula (1) are straight-chain or branched-chain. In some embodiments, the compound of formula (1) comprises a combination of C1-C10 alkyl, C2-C10 alkenyl, C3-C10 cycloalkyl, 3- to 10-membered heterocycloalkyl, aryl and heteroaryl groups.

[0046] In some embodiments of formula (1), R 1 is H, and R 2 is -C(O)N(R 3 )(R 3’ ). In some embodiments, R 1 is -C(O)N(R 3 )(R 3’ ), and R 2 is H. In some embodiments, R 3 is halo, C1-C5 alkyl or C3-C6 cycloalkyl, and R 3’ is H. In some embodiments, R 3 is methyl, ethyl, or isopropyl, and R 3’ is H.

[0047] In some embodiments of formula (1), R 1 is H, and R 2 is -R 5 or -L 1 -R 5 . In some embodiments, R 1 is -L 1 -R 5 , and R 2is H. In some embodiments, -L 1 -R 5 is, -C(O)N(R 7 )-L 2 -N(R 7 )-R 5 , -C(O)O-L 2 -OR 5 , -C(O)N(R 7 )-L 2 -N(R 7 )-C(O)O-R 5 , -C(O)O-L 2 -N(R 7 )-C(O)O-R 5 , -C(O)N(R 7 )-L 2 -N(R 8 )-C(O)N(R 8 )-R 5 , or -C(O)O-L 2 -N(R 8 )-C(O)N(R 8 )-R 5 . In some embodiments, R 5 is

Chemical formula

[0048] In some embodiments of formula (1), R1 is -C(O)O-R 5 or -R 5 wherein R 2 is H. In some embodiments, R 1 is H and R 2 is -C(O)O-R 5 or -R 5 wherein in some embodiments, R 5 is

Chemical formula

[0049] In some embodiments of formula (1), R 1 is -L 1 -R 5 wherein in some embodiments, R 1’ is -H or -L 1 -R 5 wherein in some embodiments, R 1’ is -H. In some embodiments, R 1’ is -L 1 -R 5 wherein in some embodiments, R 1 is -L 1 -R 5 wherein R 1’ is H. In some embodiments, R 1 is -L 1 -R 5 wherein R 1’ is -L 1 -R 5 wherein in some embodiments, R 1 is -L 1 -R 5 wherein R 1’is H, and R 2 is H. In some embodiments, R 1 is -L 1 -R 5 and R 1’ is -L 1 -R 5 and R 2 is H. In some embodiments, -L 1 -R 5 is (C1-C10 alkylene)-N(R 7 )-R 5 , -(C1-C10 alkylene)-O-R 5 , -C(O)O-L 2 -N(R 7 )-R 5 , -C(O)O-L 2 -O-R 5 , -C(O)O-L 2 -C(O)O-R 5 , -C(O)O-L 2 -C(O)N(R 7 )-R 5 , -C(O)N(R 7 )-R 5 , -C(O)N(R 7 )-L 2 -N(R 7 )-R 5 , -C(O)N(R 7 )-L 2 -O-R 5 , -C(O)O-L 2 -N(R 7 )C(O)O-R 5 , -C(O)N(R 8 )-L 2 -N(R 7 )C(O)O-R 5 , -C(O)O-L 2 -N(R 7 )C(O)N(R 8 )-R 5 , -C(O)N(R 7 )-L 2 -N(R 7 )C(O)N(R 8 )-R 5 , -C(O)N(R 7 )-L 2 -C(O)O-R 5and -C(O)N(R 7 )-L 2 -C(O)N(R 8 )-R 5 - is selected from the group consisting of. In some embodiments, -L 1 -R 5 is -C(O)O-R 5 、-C(O)O-L 2 -N(R 7 )-R 5 、-C(O)O-L 2 -N(R 7 )C(O)O-R 5 、-C(O)N(R 8 )-L 2 -N(R 7 )C(O)O-R 5 、-C(O)O-L 2 -N(R 7 )C(O)N(R 8 )-R 5 、-C(O)N(R 7 )-L 2 -N(R 7 )C(O)N(R 8 )-R 5 、-C(O)N(R 7 )-L 2 -N(R 7 )-R 5 and -C(O)N(R 7 )-L 2 -C(O)N(R 8 )-R 5 - is selected from the group consisting of. In some embodiments, -L 1 -R 5 is -C(O)O-R 5 。In some embodiments, R 2 is H.

[0050] Also, the divalent linker L 1 may include one or more repetitions of the same group, or a combination of different groups. In some embodiments, L 1 includes -C(O)O- and C1-C10 alkyl. In some embodiments, L 1 includes two or more repetitions of -C(O)O-. In some embodiments, L 1contains two or more repeats of -C(O)O(CH2) n -. In some embodiments, L 1 contains two or more repeats of -C(O)N(R 7 ). In some embodiments, L 1 contains two or more repeats of -C(O)N(R 7 )(CH2) n -. In some embodiments, L 1 contains a combination of -C(O)O-, C1-C10 alkyl, and -C(O)N(R 7 ). In some embodiments, L 1 contains a combination of -C(O)O-, -(CH2) n (-), and -C(O)N(R 7 ). In some embodiments, L 1 contains a combination of -C(O)N(R 7 )- and C1-C10 alkyl. In some embodiments, L 1 contains two or more repeats of -C(O)O(CH2) n -C(O)N(R 7 ). In some embodiments, n is an integer from 0 to 10. In some embodiments, n is an integer from 1 to 3. As will be understood in the art, the above combinations are non-limiting examples, and other chemically possible combinations of L 1 are also contemplated by the present disclosure.

[0051] The adenosine derivatives of the present disclosure may include one or more of their isomers. Isomers may include chiral isomers, also known as stereoisomers, which contain one or more chiral centers, amino isomers, imino isomers, or tautomers that can interconvert via the rearrangement of protons or other atoms, such as combinations thereof. In an example, the adenosine derivative can have an amino isomer, an imino isomer, or a combination thereof. In a further example, the adenosine derivative may include enantiomers, diastereomers and cis / trans isomers, tautomers, or combinations thereof. Isomers that can have reverse transcriptase inhibitor (RTI) activity in vivo may be particularly preferred in some cases.

[0052] In some embodiments of formula (1), X is a halogen atom selected from the group consisting of fluorine, chlorine, bromine, and iodine (alternatively referred to as fluoro (F), chloro (Cl), bromo (Br), and iodo (I)). In some embodiments, X is F. In some embodiments, X is Cl. In some embodiments, X is Br.

[0053] In some embodiments, the present disclosure is directed to an adenosine derivative having formula (1a), or a pharmaceutically acceptable salt, tautomer, or solvate thereof,

Chemical formula

[0054] In some embodiments, the present disclosure is directed to an adenosine derivative having formula (1b), or a pharmaceutically acceptable salt, tautomer, or solvate thereof,

Chemical formula

[0055] In some embodiments, the adenosine derivatives of the present disclosure are Formula (2):

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0056] In some embodiments, X is Cl, F, or Br. In some embodiments, X is F.

[0057] In a further embodiment, the adenosine derivatives of the present disclosure are Formula (2-A):

Chemical formula

Chemical formula

[0058] In some embodiments, the adenosine compounds of the present disclosure are isomers of Formulas (1)-(8), Formula (1a), Formula (1b), or Formulas (1-A)-(8-A). In some embodiments, the isomers are stereoisomers, such as enantiomers or diastereomers. In some embodiments, the isomers are inhibitors of reverse transcriptase having activity in vivo.

[0059] As disclosed in this specification, the adenosine derivatives of the present disclosure are ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolane-2-yl)methyl isopropylcarbamate, isopropyl (9-((2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)oxolane-2-yl)-2-fluoro-9H-purin-6-yl)carbamate, ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolane-2-yl)methyl ((5-methyl-2-oxo-1,3-dioxol-4-yl)methyl) carbonate, ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolane-2-yl)methyl methylcarbamate, 4-(((9-((2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)oxolane-2-yl)-2-fluoro-9H-purin-6-yl)aminomethyl)-5-methyl-1,3-dioxolan-2-one, ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolane-2-yl)methyl (2-(methyl((5-methyl-2-oxo-1,3-dioxol-4-yl)methyl)amino)ethyl)carbamate, [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-2-ethynyl-3-hydroxy-oxolane-2-yl]-methyl 4-[methyl-[(5-methyl-2-oxo-1,3-dioxol-4-yl)methoxycarbonyl]amino]butanoate, or a pharmaceutically acceptable salt thereof.

[0060] In some embodiments, each of the adenosine derivatives of the present disclosure independently contains one or more 5- to 10-membered heterocycles, R 1 , R 1’ , or R 2 . R 1 , R 1’ , or R 2Each can independently include, in one embodiment, a 5-membered heterocyclic ring, in another embodiment, a 6-membered heterocyclic ring, or in yet another embodiment, a 6- to 10-membered heterocyclic ring. The heterocyclic rings of the present disclosure can have one or more substituents. In some embodiments, the 5-membered heterocyclic ring includes carbon atoms and 1 to 4 heteroatoms selected from the group consisting of N, O, and S. In some embodiments, the 5-membered heterocyclic ring includes 1 to 3 O atoms. In a further embodiment, R 1 R 1’ or R 2 each independently includes a 5-membered heterocyclic ring of Formulas 9 to 24. In still further embodiments, the adenosine derivatives of the present disclosure each independently include the aforementioned -R 5 , -L 1 -R 5 , -Z-L 4 -R 5 groups, and R 1 R 1’ or R 2 . In some embodiments, -R 5 , -L 1 -R 5 , -Z-L 4 -R 5 is selected from Formulas 9 to 24 (Figures 3A to 3J and Figures 4A to 4F).

Chemical Formula

[0061] In some embodiments, -R 5 , -L 1 -R 5 , -Z-L 4 -R 5 is

Chemical Formula

[0062] In some embodiments, the adenosine derivatives of the present disclosure each have R 3 being -H, -C(O)N(R 3’ )(R 4 ) or -C(O)OR 1and R 1’ and, -C(O)N(R 3 )(R 3’ )、-C(O)OR 4 or R which is one of Formulas 9 - 24 2 and includes.

[0063] In another embodiment, the adenosine derivatives of the present disclosure each have -C(O)N(R 3 )(R 3’ ) or -C(O)OR 4 is R 1 and R 1’ and, -H, -C(O)N(R 3 )(R 3’ )、-C(O)OR 4 or R which is one of Formulas 9 - 24 2 and includes.

[0064] In some embodiments, the adenosine derivatives of the present disclosure have -H, -C(O)N(R 3 )(R 3’ ) or -C(O)OR 4 is R 2 and, -C(O)N(R 3 )(R 3’ )、or -C(O)OR 4 or R which is one of Formulas 9 - 24 1 and includes.

[0065] In yet another embodiment, the adenosine derivatives of the present disclosure have -C(O)N(R 3 )(R 3’ ) or -C(O)OR 4 is R 2 and, -H, -C(O)N(R 3 )(R 3’ )、-C(O)OR 4 or also R which is one of Formulas 9 - 24 1 and includes.

[0066] In yet another embodiment, the adenosine derivatives of the present disclosure each have, -H, -C(O)N(R 3 )(R 3’ )、-C(O)OR 4 、-R5 , -L 1 -R 5 , -Z-L 4 R 5 , or R which is one of Formulas 9-24 1 and R 1’ and, -C(O)N(R 3 )(R 3’ ), -C(O)OR 4 , -R 5 , -L 1 -R 5 , -Z-L 4 -R 5 or R which is one of Formulas 9-24 2 and includes.

[0067] In yet another embodiment, the adenosine derivative of the present disclosure is -C(O)N(R 3 )(R 3’ ), -C(O)OR 4 , -R 5 , -L 1 -R 5 , -Z-L 4 R 5 , or R which is one of Formulas 9-24 1 and, -H, -C(O)N(R 3 )(R 3’ ), -C(O)OR 4 , -R 5 , -L 1 -R 5 , -Z-L 4 -R 5 or R which is one of Formulas 9-24 2 and includes.

[0068] In still further embodiments, each of R 1 , R 1’ and R 2 is independently selected from one of Formulas 9-24.

[0069] The adenosine derivative of the present disclosure can be converted into a target drug and may include reverse transcriptase inhibitor activity in vivo, reverse transcriptase chain terminator activity in vivo, DNA translocation inhibitor activity in vivo, or a combination thereof.

[0070] The adenosine derivatives of the present disclosure may be prodrugs that do not have the activity of the original form shown herein or have limited activity and are metabolized in vivo to exhibit the desired activity of a target drug, including reverse transcriptase inhibitor activity, reverse transcriptase chain terminator activity, DNA translocation inhibitor activity, or a combination thereof.

[0071] Without wishing to be bound by any particular mechanism or theory, the applicants have discovered that the adenosine derivatives of the present disclosure can be metabolized in vivo to produce compounds or mixtures of compounds that are similar or identical to the target drug 4'-ethynyl-2-fluoro-2'-deoxyadenosine (EFdA), which has reverse transcriptase inhibitor and other antiviral activities.

[0072] As disclosed herein, the adenosine derivatives of the present disclosure can include their isomers (e.g., enantiomers, diastereomers, and / or tautomers), one or more pharmaceutically acceptable salts thereof, one or more solvates including hydrates thereof, solvate salts thereof, or mixtures thereof.

[0073] The present disclosure further relates to pharmaceutical compositions comprising the adenosine derivatives disclosed herein and a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition comprises an adenosine derivative having the formula (1), [Chemical formula] wherein, R 1 , R 1’ and R 2 are each independently -H, -C(O)N(R 3 )(R 3’ ), -C(O)OR 4 , -R 5 , -L 1 -R 5 , or Z-L 4 -R 5 ; R 1 and R 2At least one of them is not -H, but R 3 R 3’ and R 4 are each, independently, -H, C1-C10 alkyl, C2-C10 alkenyl, C3-C10 cycloalkyl, 3-10 membered heterocycloalkyl, aryl, or heteroaryl, R 5 is

Chemical formula

[0074] As disclosed above, the pharmaceutical compositions of the present disclosure containing adenosine derivatives may not contain a monophosphate group, a diphosphate group, a triphosphate group, or a combination thereof. In some embodiments, the R 1 and / or R 2 groups of the adenosine derivatives disclosed herein do not contain a monophosphate group, a diphosphate group, a triphosphate group, or a combination thereof.

[0075] In some embodiments, the C1-C10 alkyl and C2-C10 alkenyl are straight-chain or branched-chain. In some embodiments, the adenosine derivative of the pharmaceutical composition comprises a combination of C1-C10 alkyl, C2-C10 alkenyl, C3-C10 cycloalkyl, 3- to 10-membered heterocycloalkyl, aryl and heteroaryl.

[0076] In some embodiments of the pharmaceutical composition, the adenosine derivative comprises a divalent linker L comprising one or more repeats of the same groups or a combination of different groups disclosed herein. 1 comprises. Linker L 1 And non-limiting examples of other chemically possible combinations include those described above, for example, those described in formula (1).

[0077] In some embodiments, the adenosine derivative of the pharmaceutical composition is an isomer of formula (1)-(8), formula (1a), formula (1b), formula (1-A)-(8-A), formula (4-B), or formula (4-C). The above-mentioned isomers such as tautomers, enantiomers, diastereomers, cis / trans isomers, or combinations thereof may be suitable. In some embodiments, the isomer is an inhibitor of reverse transcriptase having in vivo activity.

[0078] In some embodiments, X is a halogen atom selected from the group consisting of fluorine, chlorine, bromine, and iodine (alternatively referred to as fluoro (F), chloro (Cl), bromo (Br), and iodo (I)). In one embodiment, X is F. In another embodiment, X is Cl. In yet another embodiment, X is Br. Non-limiting examples of the adenosine derivatives of the present disclosure are shown in FIGS. 1A-1I.

[0079] In some embodiments, the pharmaceutical composition of the present disclosure Formula (2):

Chemical formula

Chemical formula

Chem.

Chem.

Chem.

Chem.

Chem.

Chem.

[0080] In some embodiments, X is Cl, F, or Br. In some embodiments, X is F.

[0081] In some embodiments, the pharmaceutical composition of the present disclosure Formula (2-A):

Chem.

Chem.

Chem.

Chem.

Chem.

Chem.

Chem.

Chem.

[0082] In some embodiments, the adenosine compounds of the present disclosure are isomers of Formulas (1)-(8), Formula (1a), Formula (1b), Formulas (1-A)-(8-A), Formula (4-B), or Formula (4-C). In some embodiments, the isomers are stereoisomers, such as enantiomers or diastereomers. In some embodiments, the isomers are inhibitors of reverse transcriptase having activity in vivo.

[0083] As disclosed herein, the pharmaceutical composition of the present disclosure comprises an adenosine derivative selected from the group consisting of ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl)methyl isopropylcarbamate, isopropyl (9-((2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2-fluoro-9H-purin-6-yl)carbamate, ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl)methyl ((5-methyl-2-oxo-1,3-dioxol-4-yl)methyl)carbonate, ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl)methyl methylcarbamate, 4-(((9-((2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2-fluoro-9H-purin-6-yl)aminomethyl)-5-methyl-1,3-dioxol-2-one, ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl)methyl (2-(methyl((5-methyl-2-oxo-1,3-dioxol-4-yl)methyl)amino)ethyl)carbamate, [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl]-methyl 4-[methyl-[(5-methyl-2-oxo-1,3-dioxol-4-yl)methoxycarbonyl]amino]butanoate, and any pharmaceutically acceptable salts thereof.

[0084] In some embodiments, the pharmaceutical composition of the present disclosure comprises an adenosine derivative comprising R selected from Formulas 9-24 5 , -L 1 , -R 5 , or -Z-L 4 , -R 5 . [Chemical]

[0085] In some embodiments, the pharmaceutical composition of the present disclosure comprises an adenosine derivative comprising R and R, each of which is -H, -C(O)N(R)(R) or -C(O)OR, and -C(O)N(R)(R), -C(O)OR or R which is one of Formulas 9-24. 3 (R 3’ )(R 4 ) or -C(O)OR 1 and R 1’ and -C(O)N(R 3 (R 3’ )), -C(O)OR 4 or R which is one of Formulas 9-24. 2

[0086] In another embodiment, the pharmaceutical composition of the present disclosure comprises an adenosine derivative comprising R which is -C(O)N(R)(R) or -C(O)OR, and -H, -C(O)N(R)(R), -C(O)OR or R which is one of Formulas 9-24. 3 (R 3’ ) or -C(O)OR 4 and R 1 and -H, -C(O)N(R 3 (R 3’ )), -C(O)OR 4 or R which is one of Formulas 9-24. 2

[0087] In yet another embodiment, the pharmaceutical composition of the present disclosure comprises an adenosine derivative comprising R which is -H, -C(O)N(R)(R) or -C(O)OR, and -C(O)N(R)(R), or -C(O)OR or R which is one of Formulas 9-24. 3 (R 3’ ) or -C(O)OR 4 and R 2 and -C(O)N(R 3 (R 3’ )), or -C(O)OR 4 or R which is one of Formulas 9-24. 1

[0088] In some embodiments, the pharmaceutical composition of the present disclosure comprises R which is -C(O)N(R)(R) or -C(O)OR, and each of which is -H, -C(O)N(R 3 (R 3’ ) or -C(O)OR 4 and R 2 and -C(O)N(R 3 ​​​)(R 3’ )、 -C(O)OR 4 or R which is one of Formulas 9 - 24 1 and R 1 ’, and an adenosine derivative containing the same.

[0089] In yet another embodiment, the pharmaceutical composition of the present disclosure, each of which is -H, -C(O)N(R 3 )(R 3’ )、 -C(O)OR 4 、 -R 5 、 -L 1 -R 5 、 -Z-L 4 R 5 、 or R which is one of Formulas 9 - 24 1 and R 1’ and -C(O)N(R 3 )(R 3’ )、 -C(O)OR 4 、 -R 5 、 -L 1 -R 5 、 -Z-L 4 -R 5 or R selected from one of Formulas 9 - 24 2 and an adenosine derivative containing the same.

[0090] In yet another embodiment, the pharmaceutical composition of the present disclosure is -C(O)N(R 3 )(R 3’ )、 -C(O)OR 4 、 -R 5 、 -L 1 -R 5 、 -Z-L 4 R 5 、 or R which is one of Formulas 9 - 24 1 and -H, -C(O)N(R 3 )(R 3’ )、 -C(O)OR 4 、 -R 5 、 -L 1 -R 5 、 -Z-L 4 -R 5 or R selected from one of Formulas 9 - 24 2 and an adenosine derivative containing the same.

[0091] In still further embodiments, the pharmaceutical composition of the present disclosure comprises an adenosine derivative comprising R, each independently selected from one of Formulas 9-24 1 , R 1’ , and R 2 .

[0092] In some embodiments, R 3 , R 3’ and R 4 are each independently -H, C1-C10 alkyl, C2-C10 alkenyl, or C3-C10 cycloalkyl. In some embodiments, R 3 , R 3’ and R 4 are each independently -H, C1-C5 alkyl, C2-C5 alkenyl, or C3-C6 cycloalkyl. In some embodiments, R 3 , R 3’ and R 4 are each independently -H, or C1-C5 alkyl. In some embodiments, C1-C5 alkyl is methyl, ethyl, or isopropyl. In some embodiments 3 , R 3’ and R 4 are each independently -H, methyl, or isopropyl.

[0093] In some embodiments, R 6 is -H, C1-C10 alkyl, C2-C10 alkenyl, or C3-C10 cycloalkyl. In some embodiments, R 6 is -H, C1-C5 alkyl, C2-C5 alkenyl, or C3-C6 cycloalkyl. In some embodiments, R 6 is -H, C1-C3 alkyl, or C2-C4 alkenyl. In some embodiments, R 6 is selected from the group consisting of -H, methyl, ethyl, isopropyl, or cyclopropyl. In some embodiments, R 6 is methyl.

[0094] In some embodiments, R 7 and R 8 are each, independently, -H, C1-C10 alkyl, or C3-C6 cycloalkyl. In some embodiments, R 7 and R 8 are each, independently, -H, C1-C5 alkyl, or C3-C6 cycloalkyl. In some embodiments, R 7 and R 8 are each, independently, -H, methyl, ethyl, isopropyl, or cyclopropyl.

[0095] In some embodiments, R 9 is -H, F, C1-C10 alkyl, or C2-C10 alkenyl. In some embodiments, R 9 is -H, F, C1-C3 alkyl, or C2-C4 alkenyl. In some embodiments, R 9 is -H, F, or C1-C3 alkyl. In some embodiments, R 9 is -H.

[0096] As described, the pharmaceutical compositions of the present disclosure include a pharmaceutically acceptable carrier.

[0097] Non-limiting examples of pharmaceutically acceptable carriers include pharmaceutical excipients, surfactants, emulsifiers, fillers, carriers, isotonic agents, dispersants, viscosity modifiers, resuspending agents, buffers, or combinations thereof. Pharmaceutical excipients typically do not have the properties of a pharmaceutical or drug-active ingredient, also known as the active pharmaceutical ingredient (API), and are typically used to streamline the manufacturing process or packaging of the active ingredient, or to deliver the API to a patient or other subject. Pharmaceutically acceptable carriers, excipients or inactive ingredients from the Inactive Ingredients Database available from the US FDA (https: / / www.fda.gov / drugs / drug-approvals-and-databases / inactive-ingredients-database-download) may be suitable. Some of the generally recognized as safe (GRAS) food substances available from the US FDA's GRAS Substances (SCOGS) Database (https: / / www.fda.gov / food / generally-recognized-safe-gras / gras-substances-scogs-database) may also be suitable.

[0098] In some embodiments of the present disclosure, pharmaceutically acceptable carriers include acacia, animal oil, benzyl alcohol, benzyl benzoate, calcium stearate, carbomer, cetostearyl alcohol, cetyl alcohol, cholesterol, cyclodextrin, dextrose, diethanolamine, emulsifying wax, ethylene glycol palmitostearate, glycerin, glyceryl monostearate, glycerol stearate, glyceryl monooleate, glyceryl monostearate, hydrous, histidine, hydrochloric acid, hydroxypropyl cellulose, hydroxypropyl-β-cyclodextrin (HPBCD), hypromellose (hydroxypropyl methylcellulose (HPMC)), lanolin, lanolin alcohol, lecithin, medium-chain triglycerides, metal soaps, methylcellulose, mineral oil, monobasic sodium phosphate, monoethanolamine, oleic acid, polyethylene glycol (PEG3350, PEG4000, PEG6000), polyoxyethylene-polyoxypropylene copolymer (poloxamer), polyoxyethylene alkyl ether, polyoxyethylene castor oil, polyoxyethylene castor oil derivative, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene stearate, polysorbate, polyoxyethylene (20) sorbitan monolaurate (Tween20, Polysorbate20), polyoxyethylene (20) sorbitan monooleate (Tween80, Polysorbate80), povidone, propylene glycol alginate, physiological saline, sodium chloride, sodium citrate, sodium citrate dihydrate, sodium hydroxide, sodium lauryl sulfate, monobasic sodium acid, dibasic sodium phosphate, sorbitan ester, stearic acid, stearyl alcohol, sunflower oil, tragacanth, triethanolamine, vegetable oil, water, xanthan gum, or combinations thereof.

[0099] In a further embodiment, the pharmaceutically acceptable carrier includes dextrose, glycerin, histidine, hydrochloric acid, hydroxypropylcellulose, hydroxypropyl-β-cyclodextrin (HPBCD), hypromellose (hydroxypropylmethylcellulose (HPMC)), polyoxyethylene (20) sorbitan monolaurate (Tween20, PolysoRbate20), polyethylene glycol (PEG400, PEG3350, PEG4000, PEG6000), polyoxyethylene-polyoxypropylene copolymer (Poloxamer188, Poloxamer407), polyoxyethylene (20) sorbitan monooleate (Tween80, PolysoRbate80), physiological saline, sodium chloride, sodium citrate, sodium citrate dihydrate, sodium lauryl sulfate, monobasic sodium phosphate, dibasic sodium phosphate, or combinations thereof.

[0100] The pharmaceutical composition of the present disclosure may further include one or more anti-HIV agents (also referred to as anti-HIV drugs) selected from abacavir, abacavir sulfate, lamivudine, amprenavir, atazanavir sulfate, AZT, bictegravir, cabotegravir, darunavir, didanosine, dideoxyinosine, dolutegravir, doravirine, efavirenz, emtricitabine, tenofovir disoproxil fumarate, tenofovir alafenamide, 4'-ethynyl-2-fluoro-2'-deoxyadenosine, elvitegravir, etravirine, fosamprenavir calcium, indinavir, indinavir sulfate, lamivudine, lopinavir, the combination of lopinavir and ritonavir, darunavir, the combination of darunavir and cobicistat, maraviroc, nelfinavir, nelfinavir mesylate, nevirapine, PPL-100, raltegravir, rilpivirine, stavudine, tipranavir, vicriviroc, or combinations thereof.

[0101] In some embodiments, the pharmaceutical composition of the present disclosure can be included in a single formulation that is co-administered to a subject with a deoxynosine derivative, such as those of formulas (1)-(8), formula (1a), formula (1b), or formulas (1-A)-(8-A), and one or more anti-HIV agents. The pharmaceutical composition of the present disclosure can be included in another formulation that can be administered simultaneously or sequentially to a subject with a deoxynosine derivative and one or more anti-HIV agents. The pharmaceutical composition of the present disclosure can also be mixed in another formulation that can be co-administered to a subject with one or more anti-HIV agents.

[0102] The present disclosure further relates to a method for treating a disease. The method can include administering to a subject in need thereof a pharmaceutical composition comprising an effective dosage of a deoxynosine derivative having formula (1),

Chemical formula

Chemical formula

[0103] Any of the aforementioned adenosine derivatives (e.g., formulas (1)-(8), formula (1a), formula (1b), or formulas (1-A)-(8-A)), or any of the pharmaceutical compositions containing an adenosine derivative may be suitable. X is a halogen atom selected from the group consisting of fluorine, chlorine, bromine, and iodine (alternatively, referred to as fluoro (F), chloro (Cl), bromo (Br), and iodo (I)). In some embodiments, X is F. In another embodiment, X is Cl. In yet another embodiment, X is Br.

[0104] In some embodiments of the method, the adenosine derivative is Formula (2): [Chemical formula] Formula (3): [Chemical formula] Formula (4): [Chemical formula] Formula (5): [Chemical formula] Formula (6): [Chemical formula] Formula (7): [Chemical formula] Formula (8): [Chemical formula] Formula (4-B): [Chemical formula] comprises a formula selected from the group consisting of, its isomers, or a pharmaceutically acceptable salt thereof.

[0105] In some embodiments, X is Cl, F or Br. In some embodiments, X is F.

[0106] In some embodiments of the method, the adenosine derivative is Formula (2-A): [Chemical formula] Formula (3-A): [Chemical formula] Formula (4-A): [Chemical formula] Formula (5-A): [Chemical formula] Formula (6-A): [Chemical formula] Formula (7-A): [Chemical formula] Formula (8-A): [Chemical formula] Formula (4-C): [Chemical formula] comprises a formula selected from the group consisting of, an isomer thereof, or a pharmaceutically acceptable salt thereof.

[0107] As disclosed herein, the method comprises administering a pharmaceutical composition comprising an adenosine derivative, the adenosine derivative being ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolane-2-yl)methyl propan-2-ylcarbamate, propan-2-yl (9-((2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)oxolane-2-yl)-2-fluoro-9H-purin-6-yl)carbamate, ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolane-2-yl)methyl ((5-methyl-2-oxo-1,3-dioxol-4-yl)methyl) carbonate, ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolane-2-yl)methyl methylcarbamate, 4-(((9-((2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)oxolane-2-yl)-2-fluoro-9H-purin-6-yl)aminomethyl)-5-methyl-1,3-dioxolan-2-one, or a pharmaceutically acceptable salt thereof.

[0108] In the method of the present disclosure, the pharmaceutical composition is administered to a subject via intramuscular (IM) injection, subcutaneous (SC) injection, intravenous (IV) injection, oral administration, topical application, implant application, or a combination thereof. Implant application may include an implantable device or film containing the pharmaceutical composition disclosed herein. Implant application may include a vaginal ring, film, membrane, patch, other device, or a combination thereof.

[0109] The method of the present disclosure may further include measuring a sample of interest to determine the measured level of a target drug, where the target drug has the formula (T-1): [Chemical formula] It may have its isomers or its pharmaceutically acceptable salts. In some embodiments, X is a halogen selected from the group consisting of F, Cl, Br, and I, and in some embodiments, X is I.

[0110] In some embodiments, the target drug has the formula (T-1A): [Chemical formula] It may have its isomers or its pharmaceutically acceptable salts (Figures 5A - 5B).

[0111] In some embodiments, the target drug may be (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (also referred to as 4'-ethynyl-2-fluoro-2'-deoxyadenosine, EFdA), or its pharmaceutically acceptable salt.

[0112] In some embodiments, the target drug may be a degradation product or metabolite of compound (T-1), (T-1A), or EFdA.

[0113] The specimen can be a blood sample, urine sample, body fluid sample, tissue sample, or a combination thereof from a subject such as a patient.

[0114] The measured level of the target drug can be determined by analytical methods known to those skilled in the art, including but not limited to HPLC, GC, MS, GC-MS, or combinations thereof.

[0115] The method of the present disclosure can further include adjusting the effective dosage to yield a modified effective dosage when the level of the target drug to be measured is different from the target level of the predetermined target drug, and administering the modified effective dosage to the subject.

[0116] In some embodiments of the method, the disease is acquired immunodeficiency syndrome (AIDS), wild-type HIV-1, NRTI-resistant HIV-1, HIV-2, HIV with the M184V mutation, HIV with K65R, multi-drug resistant HIV, or an RNA virus infection.

[0117] In some embodiments, the method of the present disclosure further includes administering to the subject one or more anti-HIV agents selected from the group consisting of abacavir, abacavir sulfate, lamivudine, amprenavir, atazanavir sulfate, AZT, bictegravir, cabotegravir, darunavir, didanosine, dideoxycytidine, dolutegravir, delavirdine, efavirenz, emtricitabine, tenofovir disoproxil fumarate, tenofovir alafenamide, 4'-ethynyl-2-fluoro-2'-deoxyadenosine, elvitegravir, etravirine, fosamprenavir calcium, indinavir, indinavir sulfate, lamivudine, lopinavir, the combination of lopinavir and ritonavir, darunavir, the combination of darunavir and cobicistat, maraviroc, nelfinavir, nelfinavir mesylate, nevirapine, PPL-100, raltegravir, rilpivirine, stavudine, tipranavir, and vicriviroc, or combinations thereof. Other anti-HIV agents that have been identified or developed, or combinations thereof, may also be suitable.

[0118] The combination of the adenosine derivatives of the present disclosure (e.g., Formulas (1)-(8), Formula (1a), Formula (1b), or Formulas (1-A)-(8-A)) and one or more anti-HIV agents described herein may be useful for the treatment or prevention of AIDS or other HIV-related symptoms. The anti-HIV agents can be used in these combinations at their conventionally reported dosage ranges and regimens in the art, including, for example, the dosages described in the Physicians’ Desk Reference, Thomson PDR, Thomson PDR, 57th edition (2003), the 58th edition (2004), or the 59th edition (2005) and the current Physicians’ Desk Reference (68th ed.). (2014), Montvale, N.J.: PDR Network.

[0119] The adenosine derivatives of the present disclosure and one or more anti-HIV agents described herein can be administered to a subject together or separately via oral administration, parenteral administration, or a combination thereof. The adenosine derivatives and one or more anti-HIV agents can be administered to the subject on a daily, weekly, bi-weekly, or monthly dosing schedule.

[0120] The present disclosure is directed to the use of a pharmaceutical composition for the treatment of a disease in a subject in need of treatment of the disease, wherein the disease is acquired immunodeficiency syndrome (AIDS), wild-type HIV-1, NRTI-resistant HIV-1, HIV-2, HIV having the M184V mutation, HIV having K65R, multi-drug resistant HIV, or an RNA virus infection. Any of the aforementioned pharmaceutical compositions may be suitable. The pharmaceutical composition can be used together with one or more anti-HIV agents for the treatment of the diseases referred to herein. The adenosine derivatives and one or more anti-HIV agents can be administered to a subject together or separately via oral administration, parenteral administration, or a combination thereof. The adenosine derivatives and one or more anti-HIV agents can be administered to the subject on a daily, weekly, bi-weekly, or monthly dosing schedule.

[0121] The present disclosure further relates to the use of an adenosine derivative, optionally in combination with one or more pharmaceutically acceptable carriers disclosed herein, for the manufacture of a medicament for treating a disease, which disease is acquired immunodeficiency syndrome (AIDS), wild-type HIV-1, NRTI-resistant HIV-1, HIV-2, HIV having the M184V mutation, HIV having K65R, multi-drug resistant HIV, or an RNA virus infection. The aforementioned adenosine derivative may be suitable. The aforementioned pharmaceutically acceptable carrier may be suitable.

[0122] The present disclosure further relates to a method for preventing infection in a subject in need thereof, the method comprising administering to the subject an effective dosage of a pharmaceutical composition of the method disclosed herein, wherein the subject does not have detectable symptoms of an infectious disease. In some embodiments, the infectious disease comprises a disease selected from acquired immunodeficiency syndrome (AIDS), wild-type HIV-1, NRTI-resistant HIV-1, HIV-2, HIV having the M184V mutation, HIV having K65R, multi-drug resistant HIV, an RNA virus infection, or combinations thereof.

[0123] Detectable symptoms include, but are not limited to, symptoms of acquired immunodeficiency syndrome (AIDS), wild-type HIV-1, NRTI-resistant HIV-1, HIV-2, HIV having the M184V mutation, HIV having K65R, multi-drug resistant HIV, or combinations thereof, which are symptoms of HIV virus infection. Detection of the HIV virus can be performed by PCR, reverse transcription PCR, antigen or immuno-detection of antibodies related to AIDS or HIV.

[0124] In some embodiments, the pharmaceutical composition of the method is administered to the subject on a daily, weekly, bi-weekly or monthly dosing schedule.

[0125] In some embodiments, the method of the present disclosure further comprises administering to a subject an effective dosage of one or more anti-HIV agents selected from abacavir, abacavir sulfate, lamivudine, amprenavir, atazanavir sulfate, AZT, vicriviroc, cabotegravir, darunavir, didanosine, dideoxyinosine, dolutegravir, dolaprevir, efavirenz, emtricitabine, tenofovir disoproxil fumarate, tenofovir alafenamide, 4'-ethynyl-2-fluoro-2'-deoxyadenosine, elvitegravir, etravirine, fosamprenavir calcium, indinavir, indinavir sulfate, lamivudine, lopinavir, the combination of lopinavir and ritonavir, darunavir, the combination of darunavir and cobicistat, maraviroc, nelfinavir, nelfinavir mesylate, nevirapine, PPL-100, raltegravir, rilpivirine, stavudine, tipranavir, vicriviroc, or combinations thereof. The one or more anti-HIV agents can be administered to the subject together with, or separately from, the pharmaceutical composition of the present disclosure.

[0126] Without being bound by any particular theory, the advantage of the adenosine derivatives disclosed herein (e.g., Formulas (1)-(8), Formula (1a), Formula (1b), Formulas (1-A)-(8-A), Formula (4-B), or Formula (4-C)) is the rapid conversion to the target drug. As described below, greater than about 60% of the adenosine derivatives of the present disclosure can be surprisingly and unexpectedly converted to the target drug within about 30 minutes of contact with human plasma.

[0127] The present disclosure is illustrated by the following non-limiting examples.

Example

[0128] The present invention is further defined in the following examples. It should be understood that these examples, while showing preferred embodiments of the invention, are provided by way of illustration only. From the above discussion and these examples, one skilled in the art can ascertain the essential characteristics of the invention and make various changes and modifications to the invention to adapt it to various uses and conditions without departing from its spirit and scope.

[0129] Properties of the adenosine derivative The properties of the adenosine derivative are listed in Table 1. [Table 1]

[0130] Synthesis of the adenosine derivative prodrug: The preparation method, intermediates and synthesis schedule are outlined below.

[0131] Intermediate 1: tert-Butyl N-[9-[(2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]-2-fluoro-purin-6-yl]carbamate [Chemical formula]

[0132] Preparation of Tert-butyl N-(2-fluoro-9H-purin-6-yl)carbamate [Chemical formula]

[0133] To a stirred suspension of 2-fluoro-9H-purin-6-amine (5 g, 32.65 mmol, 1 equiv) and DMAP (399 mg, 3.27 mmol, 0.1 equiv) in THF (50 mL) was added (Boc)2O (24.94 g, 114.29 mmol, 26.3 mL, 3.5 equiv) in THF (25 mL) at 0 °C. The resulting suspension was stirred at 10 °C for 16 h, diluted with MTBE (100 mL), and quenched with water (100 mL). The organic layer was separated and washed with aqueous citric acid solution (10 wt%, 100 mL), water (2 × 100 mL), and saturated aqueous sodium chloride solution (100 mL). The organic layer was concentrated under reduced pressure to give tert-butyl N-tert-butoxycarbonyl-N-(2-fluoro-9H-purin-6-yl)carbamate (11.5 g, crude) as a yellow oil, which was used in the next reaction without further purification.

[0134] To a mixture of tert-butyl N-tert-butoxycarbonyl-N-(2-fluoro-9H-purin-6-yl)carbamate (11.5 g, 32.55 mmol, 1 equiv) in EtOH (100 mL) at 20 °C was added NaOH (2.5 M, 78.1 mL, 6 equiv) over 30 min, and the resulting mixture was stirred at 20 °C for 48 h. LCMS showed one major peak with the desired m / z. The mixture was warmed, the solvent was distilled off under reduced pressure, the aqueous solution was cooled to 0 °C, and neutralized with hydrochloric acid (1 M, 150 mL) to give a slurry. The solid was collected by filtration, dissolved in EtOAc (200 mL), and washed with water (2 × 150 mL) and saturated aqueous sodium chloride solution (150 mL). The resulting solution was concentrated under reduced pressure to give tert-butyl N-(2-fluoro-9H-purin-6-yl)carbamate (4 g, 15.80 mmol, 48.5% yield) as a white solid. The crude product was used in the next reaction without further purification. LCMS (ESI) m / z, C 10 H 12 FN5O2: calculated 253.1, found (M+H) + : 253.9; (M+Na) + : 275.9.

[0135] Preparation of [(2R,3S)-2-Ethynyl-3-(4-methylbenzoyl)oxy-5-penta-4-enoxy-tetrahydrofuran-2-yl]methyl 4-methylbenzoate [Chemical formula]

[0136] To a stirred solution of 4-methylbenzenesulfonic acid hydrate (964 mg, 5.07 mmol, 1 equiv) and penta-4-en-1-ol (480 mg, 5.58 mmol, 1.1 equiv) in toluene (28 mL) at 0 °C was added a solution of [(2R,3S)-2-ethynyl-5-hydroxy-3-(4-methylbenzoyl)oxy-tetrahydrofuran-2-yl]methyl 4-methylbenzoate (2 g, 5.07 mmol, 1 equiv) in toluene (32 mL). The reaction mixture was stirred at 0 °C for 1 h. The reaction was quenched with water (100 mL). The organic layer was separated and washed with saturated aqueous sodium bicarbonate solution (100 mL), water (100 mL) and saturated aqueous sodium chloride solution (100 mL). The organic layer was concentrated under reduced pressure. The crude product was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® silica flash column, eluting with a 0 - 8% ethyl acetate / petroleum ether gradient at 40 mL / min) to give [(2R,3S)-2-ethynyl-3-(4-methylbenzoyl)oxy-5-penta-4-enoxy-tetrahydrofuran-2-yl]methyl 4-methylbenzoate (1.38 g, 58.2% yield) as a colorless oil. LCMS (ESI) m / z, C 28 H 30 O6: calcd 462.2, found (M+Na) + : 485.1

[0137] Preparation of [(2R,3S,5R)-5-[6-(tert-Butoxycarbonylamino)-2-fluoro-purin-9-yl]-2-ethynyl-3-(4-methylbenzoyl)oxy-tetrahydrofuran-2-yl]methyl 4-methylbenzoate [Chemical formula]

[0138] A mixture of [(2R,3S)-2-ethynyl-3-(4-methylbenzoyl)oxy-5-penta-4-enoxy-tetrahydrofuran-2-yl]methyl 4-methylbenzoate (200 mg, 0.43 mmol, 1 equiv), tert-butyl N-(2-fluoro-9H-purin-6-yl)carbamate (131 mg, 0.52 mmol, 1.2 equiv) and 4 Å MS (1 g, 0.32 mmol) in MeCN (4 mL) was cooled to -25 °C. I2 (351 mg, 1.38 mmol, 3.2 equiv) was added and the resulting mixture was stirred at -25 °C for 16 h under a nitrogen atmosphere. The reaction mixture was then warmed to 0 °C and stirred at 0 °C for 2 h. The reaction was quenched with aqueous sodium sulfite solution (10 mL), diluted with water (10 mL), and then extracted with MTBE (30 mL). The obtained organic layer was washed with aqueous sodium hydrogen carbonate solution (10 mL) and then with aqueous sodium chloride solution (10 mL). The organic layer was then concentrated under reduced pressure and purified by flash silica gel chromatography (ISCO®; 4 g SepaFlash® silica flash column, eluting at 20 mL / min with a 0 - 50% ethyl acetate / petroleum ether gradient) to give [(2R,3S,5R)-5-[6-(tert-butoxycarbonylamino)-2-fluoro-purin-9-yl]-2-ethynyl-3-(4-methylbenzoyl)oxytetrahydrofuran-2-yl]methyl 4-methylbenzoate (155 mg, 50.0% yield) as a white solid. LCMS (ESI) m / z, C 35 H 32 FN5O7: calculated 629.2, found (M + H) + : 630.1

[0139] Preparation of tert-butyl N-[9-[(2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]-2-fluoro-purin-6-yl]carbamate

Chemical formula

[0140] To a mixture of [(2R,3S,5R)-5-[6-(tert-butoxycarbonylamino)-2-fluoro-purin-9-yl]-2-ethynyl-3-(4-methylbenzoyl)oxytetrahydrofuran-2-yl]methyl 4-methylbenzoate (155 mg, 0.246 mmol, 1 equiv) in THF (1 mL) was added NaOMe (133 mg, 0.739 mmol, 30%, 3 equiv) in MeOH (0.5 mL) at -25 °C, and the resulting mixture was stirred at -25 °C for 6 h. The mixture was then neutralized with AcOH (0.2 mL) and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (ISCO®; 12 g SepaFlash® silica flash column, eluting with a 0 - 5% MeOH / DCM gradient at 30 mL / min) to give tert-butyl N-[9-[(2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]-2-fluoro-purin-6-yl]carbamate (82 mg, 85.4% yield) as a white solid. LCMS (ESI) m / z, C 17 H 20 FN5O5: calculated 393.1, found (M+Na) + : 416.2.

[0141] Intermediate 2 (Method 1): (2R,3S,5R)-5-(6-Amino-2-fluoro-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol

Chemical Structure

[0142] Preparation of [(2R,3S,5R)-5-(6-Amino-2-fluoro-purin-9-yl)-2-ethynyl-3-(4-methylbenzoyl)oxy-tetrahydrofuran-2-yl]methyl 4-methylbenzoate

Chemical Structure

[0143] To a solution of (2R,3S,5R)-5-(6-((tert-butoxycarbonyl)amino)-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(((4-methylbenzoyl)oxy)methyl)tetrahydrofuran-3-yl 4-methylbenzoate (220 mg, 0.35 mmol, 1 equiv) in toluene (2.5 mL) was added TFA (0.25 mL) at 10 °C. The mixture was stirred at 10 °C for 48 h. The mixture was quenched by the addition of saturated sodium bicarbonate (10 mL) and extracted with EtOAc (2 × 10 mL). The organic layer was concentrated. The resulting residue was purified by flash silica gel chromatography (ISCO®; 4 g SepaFlash® silica flash column, eluting with a gradient of 0–100% ethyl acetate / petroleum ether at 20 mL / min) to give (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(((4-methylbenzoyl)oxy)methyl)tetrahydrofuran-3-yl 4-methylbenzoate (130 mg, 70% yield) as a white solid. C 28 H 24 LCMS (ESI) m / z for C + H

[0144] Preparation of (2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol

Chemical formula

[0145] A mixture of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(((4-methylbenzoyl)oxy)methyl)tetrahydrofuran-3-yl 4-methylbenzoate (130 mg, 0.246 mmol, 1 equiv) in THF (1 mL) was cooled to -25 °C. Then, NaOMe (133 mg, 0.737 mmol, 30% purity, 3 equiv) in MeOH (0.5 mL) was added and the mixture was stirred at -25 °C for 3 h. The mixture was neutralized with AcOH (0.2 mL) and the mixture was concentrated. The resulting residue was purified by flash silica gel chromatography (ISCO®; 4 g SepaFlash® silica flash column, eluting at 20 mL / min with a 0 - 8% MeOH / DCM gradient) to give (2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (46 mg, 64% yield) as a white solid. C 12 H 12 LCMS (ESI) m / z for C₉H₁₀FN₅O₃: calculated 293.2, found (M + H) + : 294.1. 1 H NMR (DMSO-d6, 400 MHz) δ (ppm) 8.30 (s, 1H), 7.89 (br s, 2H), 6.24 (dd, J = 7.2, 5.2 Hz, 1H), 5.58 (d, J = 5.6 Hz, 1H), 5.30 (t, J = 6.4 Hz, 1H), 4.52 - 4.61 (m, 1H), 3.62 - 3.68 (m, 1H), 3.55 (dd, J = 12.0, 6.8 Hz, 1H), 3.52 (s, 1H), 2.64 - 2.75 (m, 1H), 2.37 - 2.47 (m, 1H). 19 F NMR (DMSO-d6, 376 MHz) δ (ppm) -51.98 (br s, 1F).

[0146] Intermediate 2 (Method 2): (2R,3S,5R)-5-(6-Amino-2-fluoro-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol

Chem.

[0147] Preparation of (2R,3S,5R)-2-ethynyl-5-(2-fluoro-6-((trimethylsilyl)amino)-9H-purin-9-yl)-2-(((4-methylbenzoyl)oxy)methyl)tetrahydrofuran-3-yl 4-methylbenzoate

Chem.

[0148] CaH2 was added to acetonitrile and refluxed for 2 hours, then evaporated under nitrogen protection to obtain anhydrous CH3CN.

[0149] To a solution of 2-fluoro-9H-purin-6-amine (4.39 g, 28.64 mmol, 1.25 eq) in anhydrous CH3CN (60 mL) was added BSA (13.98 g, 68.74 mmol, 16.99 mL, 3 eq) under N2. The mixture was stirred at 80 °C for 1 h. After cooling the reaction mixture to 25 °C, TMSOTf (6.11 g, 27.49 mmol, 4.97 mL, 1.2 eq) was added and the reaction mixture was stirred at 25 °C for 1 h, then [(2R,3S)-5-acetoxy-2-ethynyl-3-(4-methylbenzoyl)oxy-tetrahydrofuran-2-yl]methyl 4-methylbenzoate (10 g, 22.91 mmol, 1 eq) in anhydrous CH3CN (60 mL) was added over 1 h at 80 °C and the mixture was stirred at 80 °C for 16 h under N2. The reaction mixture was cooled to room temperature (15 °C) and allowed to stand for 24 h. The slurry was filtered, washed with cold water anhydrous MeCN (10 mL), and the mixture was dried under vacuum to give [(2R,3S,5R)-2-ethynyl-5-[2-fluoro-6-(trimethylsilylamino)purin-9-yl]-3-(4-methylbenzoyl)oxy-tetrahydrofuran-2-yl]methyl 4-methylbenzoate (6.58 g, yield 47.7%) as a white solid. 1 H NMR (400 MHz, CDCl3) δ (ppm) 8.02 (d, J = 8.0 Hz, 2H), 7.93 (d, J = 8.0 Hz, 2H), 7.86 (s, 1H), 7.29 (d, J = 8.0 Hz, 2H), 7.23 (d, J = 8.0 Hz, 2H), 6.51 (t, J = 12 Hz, 1H), 6.05 (m, 1H), 4.82 (d, J = 20 Hz, 1H), 4.67 (d, J = 20 Hz, 1H), 3.23 - 3.19 (m, 1H), 2.90 - 2.85 (m, 1H), 2.68 (s, 1H), 2.43 (d, J = 16 Hz, 6H), 0.49 - 0.29 (m, 9H). 19 F NMR (376MHz, CDCl3) δ (ppm) -49.34 (s, 1F).

[0150] (2R,3S,5R)-5-(6-Amino-2-fluoro-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol Preparation

Chemical formula

[0151] To a solution of [(2R,3S,5R)-2-ethynyl-5-[2-fluoro-6-(trimethylsilylamino)purin-9-yl]-3-(4-methylbenzoyl)oxy-tetrahydrofuran-2-yl]methyl 4-methylbenzoate (15.5 g, 25.76 mmol, 1 equiv) in THF (150 mL) was slowly added NaOMe (6.96 g, 38.64 mmol, 30% purity, 1.5 equiv) at -25 °C, and the reaction mixture was stirred at -25 °C for 16 h. The mixture was neutralized to pH ~7 with AcOH (0.5 mL), and the mixture was concentrated. The crude product was triturated with CH3CN:H2O = 7:3 (100 mL) at 5 °C for 16 h to give (2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (6.76 g, 89%) as a white solid. C 12 H 12 LCMS(ESI) m / z for C + H 1 FN5O3: calculated 293.2, found (M+H) 19 : 294.1. 1 1 1 Intermediate 3: (2R,3S,5R)-5-(6-Amino-2-fluoro-purin-9-yl)-4,4-dideuterio-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol

Chem.

[0153] Preparation of tert-Butyl 2,2,2-trideuterioacetate

Chem.

[0154] To a mixture of 2-methylpropan-2-ol (40 g, 539.66 mmol, 51.61 mL, 1 equiv) and dideuterio 2,2,2-trideuterioacetate (51.87 g, 809.49 mmol, 1.5 equiv) in DMF (700 mL) were added DCC (167.02 g, 809.49 mmol, 1.5 equiv) and DMAP (13.19 g, 107.93 mmol, 0.2 equiv), and the mixture was stirred at 25 °C for 64 h. The reaction mixture was distilled under vacuum (105 °C, 0.1 MPa) to give tert-butyl 2,2,2-trideuterioacetate (25.3 g, yield 39.3%) as a colorless liquid. 1 1H NMR (400 MHz, CDCl3) δ (ppm) 1.45 (s, 9H).

[0155] Preparation of (4R)-2,2-Dimethyl-4-(2-triisopropylsilylethynyl)-1,3-dioxolane-4-carbaldehyde

Chem.

[0156] A solution of [(4S)-2,2-dimethyl-4-(2-triisopropylsilylethynyl)-1,3-dioxolan-4-yl]methanol (40.2 g, 128.63 mmol, 1 equiv) in MeCN (220 mL) was added to IBX (108.06 g, 385.90 mmol, 3 equiv). The mixture was stirred at 85 °C for 2 h. The mixture was filtered and concentrated to give (4R)-2,2-dimethyl-4-(2-triisopropylsilylethynyl)-1,3-dioxolane-4-carbaldehyde (40 g, crude) as a yellow oil.

[0157] Preparation of tert-butyl (3S)-2,2-dideuterio-3-[(4R)-2,2-dimethyl-4-(2-triisopropylsilylethynyl)-1,3-dioxolan-4-yl]-3-hydroxypropanoate

Chemical Structure

[0158] To a solution of THF (45 mL), LDA (2 M in THF, 24.15 mL, 1.5 equiv) was added at -78 °C, and tert-butyl 2,2,2-trideuterioacetate (6.14 g, 51.53 mmol, 1.6 equiv) was added at -78 °C. The mixture was stirred at -78 °C for 1 h, and (4R)-2,2-dimethyl-4-(2-triisopropylsilylethynyl)-1,3-dioxolane-4-carbaldehyde (10 g, 32.21 mmol, 1 equiv) in THF (30 mL) was added at -78 °C. Then, the mixture was stirred at -78 °C for 2 h. D2O (20 mL) was added, and then the mixture was extracted with ethyl acetate (200 mL × 2). The combined organic layers were washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated. The obtained residue was purified by flash silica gel chromatography (ISCO (registered trademark); 330 g SepaFlash (registered trademark) silica flash column, eluting with a solvent having a 0 - 70% DCM / petroleum ether gradient at 75 mL / min) to give tert-butyl (3S)-2,2-dideuterio-3-[(4R)-2,2-dimethyl-4-(2-triisopropylsilylethynyl)-1,3-dioxolan-4-yl]-3-hydroxypropanoate (5.17 g, yield 37.5%) as a colorless oil. 1 H NMR (400 MHz, CDCl3) δ (ppm) 4.22 (d, J = 8.8 Hz, 1H), 4.11 (d, J = 8.4 Hz, 1H), 4.01 (d, J = 4.8 Hz, 1H), 2.71 (d, J = 5.2 Hz, 1H), 1.60 (s, 3H), 1.48 (s, 9H), 1.42 (s, 3H), 1.08 (s, 21H).

[0159] Preparation of tert-butyl (3S)-2,2-dideuterio-3-[(4R)-4-ethynyl-2,2-dimethyl-1,3-dioxolan-4-yl]-3-hydroxy-propanoate

Chemical formula

[0160] A solution of tert-butyl (3S)-2,2-dideuterio-3-[(4R)-2,2-dimethyl-4-(2-triisopropylsilylethynyl)-1,3-dioxolan-4-yl]-3-hydroxypropanoate (13 g, 30.33 mmol, 1 equiv) in THF (130 mL) was added dropwise with TBAF (1 M in THF, 30.33 mL, 1 equiv) at 0 °C. The mixture was stirred at 25 °C for 1 h. Ethyl acetate (100 mL) and H2O (100 mL) were added, and then the mixture was extracted with ethyl acetate (100 mL × 2). The combined organic layers were washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated. The resulting residue was purified by flash silica gel chromatography (ISCO (registered trademark); 120 g SepaFlash (registered trademark) silica flash column, eluting at 60 mL / min with a 0 - 15% ethyl acetate / petroleum ether gradient), to give tert-butyl (3S)-2,2-dideuterio-3-[(4R)-4-ethynyl-2,2-dimethyl-1,3-dioxolan-4-yl]-3-hydroxypropanoate (6.7 g, yield 81.1%) as a pale yellow oil. 1 H NMR (400 MHz, CDCl3) δ(ppm) 4.26 (d, J = 8.8 Hz, 1H), 4.16 (d, J = 8.4 Hz, 1H), 3.96 (d, J = 4.0 Hz, 1H), 3.24 - 3.20 (m, 1H), 2.55 (s, 1H), 1.62 (s, 3H), 1.50 (s, 9H), 1.42 (s, 3H).

[0161] Preparation of [(1S)-3-tert-butoxy-2,2-dideuterio-1-[(4R)-4-ethynyl-2,2-dimethyl-1,3-dioxolan-4-yl]-3-oxopropyl] 4-methylbenzoate

Chemical Structure

[0162] To a solution of 4-methylbenzoyl chloride (5.71 g, 36.90 mmol, 4.88 mL, 1.5 eq) in pyridine (60 mL) was added tert-butyl (3S)-2,2-dideuterio-3-[(4R)-4-ethynyl-2,2-dimethyl-1,3-dioxolan-4-yl]-3-hydroxy-propanoate (6.7 g, 24.60 mmol, 1 eq). The mixture was stirred at 0 °C for 16 h. After adding ice water (15 mL), the mixture was stirred at 0 °C for 15 min. The mixture was filtered. The resulting residue was purified by flash silica gel chromatography (ISCO (registered trademark); 80 g SepaFlash (registered trademark) silica flash column, eluting at 60 mL / min with a 0 - 100% ethyl acetate / petroleum ether gradient) to give [(1S)-3-tert-butoxy-2,2-dideuterio-1-[(4R)-4-ethynyl-2,2-dimethyl-1,3-dioxolan-4-yl]-3-oxopropyl] 4-methylbenzoate (9.2 g, 95.8% yield) as a colorless oil.

[0163] Preparation of [(2R,3S)-4,4-dideuterio 2-ethynyl-2-(hydroxymethyl)-5-oxo-tetrahydrofuran-3-yl] 4-methylbenzoate

Chemical formula

[0164] A solution of [(1S)-3-tert-butoxy-2,2-dideuterio-1-[(4R)-4-ethynyl-2,2-dimethyl-1,3-dioxolan-4-yl]-3-oxopropyl] 4-methylbenzoate (9.2 g, 23.56 mmol, 1 equiv) in DME (90 mL) was added to HCl (conc.) (7.16 g, 70.68 mmol, 7.02 mL, 36% purity, 3 equiv). The mixture was stirred at 50 °C for 16 h. The mixture was concentrated. The resulting residue was purified by flash silica gel chromatography (ISCO®; 25 g SepaFlash® silica flash column, eluting at 30 mL / min with a 0 - 30% ethyl acetate / petroleum ether gradient) to give [(2R,3S)-4,4-dideuterio-2-ethynyl-2-(hydroxymethyl)-5-oxo-tetrahydrofuran-3-yl] 4-methylbenzoate (6.4 g, 98.3% yield) as a colorless oil.

[0165] Preparation of [(2R,3S)-4,4-dideuterio-2-ethynyl-3-(4-methylbenzoyl)oxy-5-oxo-tetrahydrofuran-2-yl]methyl 4-methylbenzoate

Chemical formula

[0166] To a mixture of [(2R,3S)-4,4-dideuterio-2-ethynyl-2-(hydroxymethyl)-5-oxo-tetrahydrofuran-3-yl] 4-methylbenzoate (2 g, 7.24 mmol, 1 equiv) in pyridine (15 mL) was added 4-methylbenzoyl chloride (1.68 g, 10.86 mmol, 1.5 equiv). The resulting mixture was stirred at 0 °C for 16 h. Water (50 mL) was added at 0 °C and the mixture was extracted with DCM (2 × 100 mL). The combined organic layers were dried over Na2SO4, filtered, and concentrated. The resulting residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash® silica flash column, eluting at 60 mL / min with a gradient of 0 - 15% ethyl acetate / petroleum ether) to afford [(2R,3S)-4,4-dideuterio-2-ethynyl-3-(4-methylbenzoyl)oxy-5-oxo-tetrahydrofuran-2-yl]methyl 4-methylbenzoate (2.5 g, 87.6% yield) as a white solid.

[0167] Preparation of [(2R,3S)-4,4-dideuterio-2-ethynyl-5-hydroxy-3-(4-methylbenzoyl)oxy-tetrahydrofuran-2-yl]methyl 4-methylbenzoate

Chemical formula

[0168] A solution of [(2R,3S)-4,4-dideuterio-2-ethynyl-3-(4-methylbenzoyl)oxy-5-oxo-tetrahydrofuran-2-yl]methyl 4-methylbenzoate (2.5 g, 6.34 mmol, 1 equiv) in DCM (16 mL) and toluene (36 mL) was added dropwise with sodium bis(2-methoxyethoxy)aluminum hydride (2.75 g, 9.51 mmol, 2.64 mL, 70% purity, 1.5 equiv) at -78 °C. The mixture was stirred at -78 °C for 2 h. The reaction was quenched by adding a solution of acetic acid (5 mL) in DCM (20 mL) over 10 min. The reaction mixture was diluted with H2O (50 mL), extracted with DCM (50×3 mL), the combined organic layers were washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated to give [(2R,3S)-4,4-dideuterio-2-ethynyl-5-hydroxy-3-(4-methylbenzoyl)oxy-tetrahydrofuran-2-yl]methyl 4-methylbenzoate (2.5 g, 99.5% yield) as a pale yellow liquid.

[0169] Preparation of [(2R,3S)-5-acetoxy-4,4-dideuterio-2-ethynyl-3-(4-methylbenzoyl)oxy-tetrahydrofuran-2-yl]methyl 4-methylbenzoate

Chemical formula

[0170] A mixture of [(2R,3S)-4,4-dideuterio-2-ethynyl-5-hydroxy-3-(4-methylbenzoyl)oxy-tetrahydrofuran-2-yl]methyl 4-methylbenzoate (2.5 g, 6.31 mmol, 1 equiv) in toluene (25 mL) was added with 4-(dimethylamino)pyridine (131 mg, 1.07 mmol, 0.17 equiv) and TEA (747 mg, 7.38 mmol, 1.03 mL, 1.17 equiv) at 0 °C, and then acetic anhydride (644 mg, 6.31 mmol, 0.60 mL, 1 equiv) was added. The resulting mixture was stirred at 0 °C for 2 h. The reaction mixture was quenched with H2O (10 mL). The resulting mixture was warmed to ambient temperature and washed successively with aqueous citric acid solution (10 wt%, 30 mL), aqueous sodium hydrogen carbonate solution (5 wt%, 30 mL) and water (30 mL), and the organic layer was concentrated. The resulting residue was purified by flash silica gel chromatography (ISCO (registered trademark); 40 g of SepaFlash (registered trademark) silica flash column, eluent having a 0-20% ethyl acetate / petroleum ether gradient at 30 mL / min) to give [(2R,3S)-5-acetoxy-4,4-dideuterio-2-ethynyl-3-(4-methylbenzoyl)oxy-tetrahydrofuran-2-yl]methyl 4-methylbenzoate (2.2 g, yield 79.6%) as a colorless oil.

[0171] Preparation of [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-4,4-dideuterio 2-ethynyl-3-(4-methylbenzoyl)oxy-tetrahydrofuran-2-yl]methyl 4-methylbenzoate

Chemical Structure

[0172] To a solution of 2-fluoro-9H-purin-6-amine (960 mg, 6.27 mmol, 1.25 equiv) in MeCN (20 mL) was added bis(trimethylsilyl)acetamide (3.06 g, 15.05 mmol, 3 equiv). The mixture was stirred at 80 °C for 1 h. When the reaction mixture was cooled to 25 °C, TMSOTf (1.34 g, 6.02 mmol, 1.09 mL, 1.2 equiv) was added and the reaction was stirred at 25 °C for 1 h, then [(2R,3S)-5-acetoxy-4,4-dideuterio-2-ethynyl-3-(4-methylbenzoyl)oxy-tetrahydrofuran-2-yl]methyl 4-methylbenzoate (2.2 g, 5.02 mmol, 1 equiv) in MeCN (15 mL) was added over 1 h and the mixture was stirred at 80 °C for 16 h. DCM (50 mL) and H2O (50 mL) were added and then the mixture was extracted with DCM (50 × 2 mL). The combined organic layers were washed with brine (50 mL), dried over Na2SO4, filtered and concentrated. The residue obtained was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® silica flash column, eluent having 0 - 40% ethyl acetate / DCM at 30 mL / min) to give [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-4,4-dideuterio-2-ethynyl-3-(4-methylbenzoyl)oxy-tetrahydrofuran-2-yl]methyl 4-methylbenzoate (1.43 g, 53.6% yield) as a pale yellow solid.

[0173] Preparation of (2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-4,4-dideuterio 2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol [Chemical formula]

[0174] A mixture of [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-4,4-dideuterio-2-ethynyl-3-(4-methylbenzoyl)oxy-tetrahydrofuran-2-yl]methyl 4-methylbenzoate (1.4 g, 2.63 mmol, 1 equiv) in THF (16 mL) was added to NaOMe (1.42 g, 7.90 mmol, 30% purity, 3 equiv) in MeOH (8 mL). The mixture was stirred at -25 °C for 3 h. The mixture was neutralized with AcOH (0.5 mL) and the mixture was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 12 g SepaFlash® silica flash column, eluent with a 0 - 10% methanol / dichloromethane gradient at 30 mL / min) to give (2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-4,4-dideuterio-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (710 mg, 91.3% yield) as a white solid.

[0175] Purification of (2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-4,4-dideuterio-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol

Chem.

[0176] (2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-4,4-dideuterio-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (20 mg, 0.068 mmol, 1 equiv) was purified by preparative TLC (SiO2, DCM:MeOH = 10:1) to give (2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-4,4-dideuterio-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (6.2 mg, 31.0% yield) as a white solid. LCMS (ESI) m / z, C 12 H 10D2FN5O3: Calculated value 295.10, measured value (M+H) + : 296.1 1 H NMR (400 MHz, CD3CN) δ ppm 8.01 (s, 1H), 6.40 - 6.31 (m, 3H), 4.67 (d, J = 4.8 Hz, 1H), 4.40 - 4.37 (m, 1H), 3.84 - 3.72 (m, 1H), 3.56 (d, J = 5.2 Hz, 1H), 2.96 (s, 1H). 19 F NMR (376 MHz, CD3CN) δ ppm -53.40 (s, 1F).

[0177] Intermediate 4 (2R,3S,5R)-5-(6-Amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl-d2)tetrahydrofuran-4,4-d2-3-ol

Chemical Structure

[0178] Preparation of 9-[(2R,4S,5R)-4-[tert-Butyl(dimethyl)silyl]oxy-5-[[tert-Butyl(dimethyl)silyl]oxymethyl]-3,3-dideuterio-5-ethynyl-tetrahydrofuran-2-yl]-2-fluoro-purin-6-amine

Chemical Structure

[0179] To a mixture of (2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-4,4-dideuterio-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (690 mg, 2.34 mmol, 1 equiv) in DMF (3 mL) was added TBSCl (1.41 g, 9.36 mmol, 1.15 mL, 4 equiv) and 1H-imidazole (956 mg, 14.04 mmol, 6 equiv). The mixture was stirred at 25 °C for 16 h. The mixture was concentrated, diluted with H2O (30 mL), and extracted with EtOAc (30 × 3 mL). The combined organic layers were washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by flash silica gel chromatography (ISCO®; 4 g SepaFlash® silica flash column, eluting at 20 mL / min with a 0–10% methanol / dichloromethane gradient) to give 9-[(2R,4S,5R)-4-[tert-butyl(dimethyl)silyl]oxy-5-[[tert-butyl(dimethyl)silyl]oxymethyl]-3,3-dideuterio-5-ethynyl-tetrahydrofuran-2-yl]-2-fluoro-purin-6-amine (1.1 g, 89.8% yield) as a white solid. C 24 H 38 LCMS (ESI) m / z for D2FN5O3Si2: calcd 523.3, found (M + H) + : 524.6.

[0180] Preparation of [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-3-[tert-butyl(dimethyl)silyl]oxy-4,4-dideuterio-2-ethynyl-tetrahydrofuran-2-yl]methanol

Chem.

[0181] A mixture of 9-[(2R,4S,5R)-4-[tert-butyl(dimethyl)silyl]oxy-5-[[tert-butyl(dimethyl)silyl]oxymethyl]-3,3-dideuterio-5-ethynyl-tetrahydrofuran-2-yl]-2-fluoro-purin-6-amine (1.1 g, 2.10 mmol, 1 equiv) in THF (12 mL), H2O (3 mL), and TFA (3 mL). The reaction mixture was stirred at 0 °C for 16 h. The resulting mixture was concentrated. The residue was purified by flash silica gel chromatography (ISCO®; 12 g SepaFlash® silica flash column, eluting with a 0 - 10% ethyl acetate / petroleum ether gradient at 30 mL / min) to give [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-3-[tert-butyl(dimethyl)silyl]oxy-4,4-dideuterio-2-ethynyl-tetrahydrofuran-2-yl]methanol (590 mg, 68.6% yield) as a white solid. C 18 H 24 D2FN5O 3 LCMS (ESI) m / z for Si: calculated 409.19, found (M + H) + : 410.1.

[0182] Preparation of (2S,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-3-[tert-butyl(dimethyl)silyl]oxy-4,4-dideuterio-2-ethynyl-tetrahydrofuran-2-carboxylic acid

Chemical Structure

[0183] To a mixture of [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-3-[tert-butyl(dimethyl)silyl]oxy-4,4-dideuterio-2-ethynyl-tetrahydrofuran-2-yl]methanol (590 mg, 1.44 mmol, 1 equiv) in H2O (0.05 mL), TEMPO (45 mg, 0.29 mmol, 0.2 equiv) and PhI(OAc)2 (557 mg, 1.73 mmol, 1.2 equiv) were added. After the reaction mixture was stirred at 25 °C for 2 h, DCM (10 mL) and CH3CN (0.05 mL) were added. The resulting mixture was stirred at 25 °C for 14 h. The reaction mixture was filtered, and the cake was washed with petroleum ether (5 mL) and then concentrated to give (2S,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-3-[tert-butyl(dimethyl)silyl]oxy-4,4-dideuterio-2-ethynyl-tetrahydrofuran-2-carboxylic acid (330 mg, yield 54.1%) as a white solid. 1 1H NMR (400 MHz, DMSO-d6) δ (ppm) 8.30 (s, 1H), 7.87 (s, 2H), 6.34 (s, 1H), 4.94 (s, 1H), 3.66 (s, 1H), 0.92 (s, 9H), 0.13 - 0.12 (d, J = 4 Hz, 6H).

[0184] Preparation of methyl (2S,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-3-[tert-butyl(dimethyl)silyl]oxy-4,4-dideuterio-2-ethynyl-tetrahydrofuran-2-carboxylate

Chemical Structure

[0185] To a mixture of (2S,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-3-[tert-butyl(dimethyl)silyl]oxy-4,4-dideuterio-2-ethynyl-tetrahydrofuran-2-carboxylic acid (60 mg, 0.14 mmol, 1 equiv) in toluene (4 mL) and MeOH (3.2 mL) was added TMSCHN2 (2.0 M, 0.22 mL, 3 equiv). The reaction mixture was stirred at 25 °C for 2 h. The reaction mixture was concentrated. The resulting residue was purified by flash silica gel chromatography (ISCO®; 4 g SepaFlash® silica flash column, eluting at 20 mL / min with a 0 - 10% methanol / dichloromethane gradient), to give methyl (2S,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-3-[tert-butyl(dimethyl)silyl]oxy-4,4-dideuterio-2-ethynyl-tetrahydrofuran-2-carboxylate (41 mg, 67% yield) as a white solid. C 19 H 24 D2FN5O 4 LCMS (ESI) m / z for Si: calculated 437.54, found (M+H) + : 438.5. 1 H NMR (400 MHz, CDCl3) δ (ppm) 8.20 (s, 1H), 6.52 (s, 1H), 5.81 (s, 2H), 4.88 (s, 1H), 3.86 (s, 3H), 2.70 (s, 1H), 0.96 (s, 9H), 0.18 - 0.17 (d, J = 4 Hz, 6H). 19 F NMR (376 MHz, CDCl3) δ (ppm) -50.588 (s, 1F).

[0186] Preparation of [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-3-[tert-butyl(dimethyl)silyl]oxy-4,4-dideuterio-2-ethynyl-tetrahydrofuran-2-yl]-dideuterio-methanol

Chem.

[0187] To a mixture of methyl (2S,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-3-[tert-butyl(dimethyl)silyl]oxy-4,4-dideuterio-2-ethynyl-tetrahydrofuran-2-carboxylate (150 mg, 0.34 mmol, 1 equiv) in i-PrOH (8 mL) was added sodium borodeuteride (29 mg, 0.68 mmol, 2 equiv) at 0 °C. The reaction mixture was stirred at 25 °C for 3 h. The reaction mixture was concentrated. The resulting residue was purified by flash silica gel chromatography (ISCO®; 4 g SepaFlash® silica flash column, eluting with a 0–10% methanol / dichloromethane gradient at 20 mL / min) to give [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-3-[tert-butyl(dimethyl)silyl]oxy-4,4-dideuterio-2-ethynyl-tetrahydrofuran-2-yl]-dideuterio-methanol (107 mg, 76.4% yield) as a white solid. 1 H NMR (400 MHz, CD3CN) δ (ppm) 7.98 (s, 1H), 6.38 (s, 2H), 6.27 (s, 1H), 4.78 (s, 1H), 4.28 (s, 1H), 2.86 (s, 1H), 0.94 (s, 9H), 0.15 - 0.14 (d, J = 4 Hz, 6H). 19 F NMR (376MHz, CD3CN) δ (ppm) -53.40 (s, 1F).

[0188] Preparation of (2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-4,4-dideuterio-2-[dideuterio(hydroxy)methyl]-2-ethynyl-tetrahydrofuran-3-ol

Chem.

[0189] A mixture of [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-3-[tert-butyl(dimethyl)silyl]oxy-4,4-dideuterio-2-ethynyl-tetrahydrofuran-2-yl]-dideuterio-methanol (180 mg, 0.437 mmol, 1 equiv) in pyridine (2.34 g, 29.28 mmol, 2.38 mL, 67 equiv) and Et3N (1.72 g, 17.04 mmol, 2.37 mL, 39 equiv) was added with N,N-diethylethanamine; trihydrofluoride (4.68 g, 29.06 mmol, 4.74 mL, 66.5 equiv) at 0 °C. The resulting mixture was stirred at 25 °C for 2 h. The reaction mixture was concentrated. The obtained residue was purified by flash silica gel chromatography (ISCO®; 4 g SepaFlash® silica flash column, eluting with a 0 - 10% methanol / dichloromethane gradient at 20 mL / min) to give (2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-4,4-dideuterio-2-[dideuterio(hydroxy)methyl]-2-ethynyl-tetrahydrofuran-3-ol (80 mg, 61.5% yield) as a white solid. C 12 H8D4FN5O 3 LCMS (ESI) m / z for: calculated 297.12, found (M + H) + : 298.1. 1 H NMR (400 MHz, DMSO-d6) δ (ppm) 8.30 (s, 1H), 7.85 (s, 2H), 6.23 (s, 1H), 5.57 - 5.55 (d, J = 8 Hz, 1H), 5.25 (s, 1H), 4.56 - 4.55 (d, J = 4 Hz, 1H), 3.51 (s, 1H). 19 F NMR (376 MHz, DMSO-d6) δ (ppm) -51.98 (s, 1F).

[0190] Example 1: ((2R,3S,5R)-5-(6-Amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl)methyl isopropylcarbamate [Chemical formula]

[0191] Preparation of [(2R,3S,5R)-5-[6-(tert-Butoxycarbonylamino)-2-fluoro-purin-9-yl]-2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl]methyl (4-nitrophenyl) carbonate [Chemical formula]

[0192] To a solution of tert-Butyl N-[9-[(2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]-2-fluoro-purin-6-yl]carbamate (80 mg, 0.203 mmol, 1 equiv) in pyridine (0.8 mL) at 10 °C was added (4-nitrophenyl) carbonochloridate (41 mg, 0.203 mmol, 1 equiv) at 10 °C. The mixture was stirred at 10 °C for 16 h, water (10 mL) was added, and the mixture was extracted with EtOAc (10 mL). The organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure to give the crude product [(2R,3S,5R)-5-[6-(tert-Butoxycarbonylamino)-2-fluoro-purin-9-yl]-2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl]methyl (4-nitrophenyl) carbonate (114 mg, crude) as a yellow oil and was used in the next reaction without further purification. LCMS (ESI) m / z, C 24 H 23 FN6O9: Calculated 558.2, found (M+H) + : 559.1.

[0193] Preparation of Tert-Butyl N-[9-[(2R,4S,5R)-5-ethynyl-4-hydroxy-5-(isopropylcarbamoyloxymethyl)tetrahydrofuran-2-yl]-2-fluoropurin-6-yl]carbamate [Chemical formula]

[0194] To a mixture of [(2R,3S,5R)-5-[6-(tert-butoxycarbonylamino)-2-fluoro-purin-9-yl]-2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl]methyl (4-nitrophenyl) carbonate (10 mg, 0.018 mmol, 1 equiv) and triethylamine (3.6 mg, 0.035 mmol, 2 equiv) in THF (0.5 mL), propan-2-amine (1.3 mg, 0.021 mmol, 1.2 equiv) was added. The mixture was stirred at 15 °C for 2.5 h, water (5 mL) was added, and the mixture was extracted with EtOAc (2 × 10 mL). The organic layer was concentrated under reduced pressure. The crude product was purified by preparative HPLC (column: Waters Xbridge Prep OBD C18 150*30 10u; mobile phase: [water (10 mM NH4HCO3)-ACN]; B%: 15% - 45%, 11 min) to give tert-butyl N-[9-[(2R,4S,5R)-5-ethynyl-4-hydroxy-5-(isopropylcarbamoyloxymethyl)tetrahydrofuran-2-yl]-2-fluoropurin-6-yl]carbamate (3.6 mg, yield 45.0%) as a white solid. LCMS (ESI) m / z, C 21 H 27 FN6O6: calculated 478.2, found (M + H) + : 479.3; (M + Na) + : 501.2.

[0195] Preparation of ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl)methyl isopropylcarbamate

Chemical formula

[0196] A solution of tert-butyl N-[9-[(2R,4S,5R)-5-ethynyl-4-hydroxy-5-(isopropylcarbamoyloxymethyl)tetrahydrofuran-2-yl]-2-fluoro-purin-6-yl]carbamate (3.6 mg, 0.0075 mmol, 1 equiv) in DCM (0.5 mL) was added with TFA (77 mg, 0.68 mmol, 0.05 mL, 89.8 equiv) at 10 °C. The mixture was stirred at 10 °C for 40 h. The mixture was concentrated under reduced pressure and purified by preparative HPLC (column: Agela DuraShell 150 mm×25 mm×5um; mobile phase: [water (0.05% HCl)-ACN], B%: 10% - 40%, 8 min) to obtain (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl)methyl isopropylcarbamate (1.2 mg, yield 40.0%) as a white solid. LCMS (ESI) m / z, C 16 H 19 FN6O4: calculated value 378.2, measured value (M + H) + : 379.3; (M + Na) + : 401.2. 1 H NMR (400 MHz, DMSO-d6) δ (ppm) 8.27 (s, 1H), 7.85 (br s, 2H), 7.13 (br d, J = 4.8 Hz, 1H), 6.24 (dd, J = 7.6, 5.2 Hz, 1H), 4.55 (br t, J = 6.8 Hz, 1H), 4.35 (br d, J = 11.6 Hz, 1H), 4.00 (br d, J = 11.6 Hz, 1H), 3.61 (s, 1H), 2.70 - 2.79 (m, 1H), 2.40 - 2.43 (m, 1H), 0.98 - 1.07 (m, 7H). 19 F NMR (376 MHz, DMSO-d6) δ (ppm) -51.79 (s).

[0197] Example 2: ((2R,3S,5R)-5-(6-Amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl)methyl carbamate Preparation of ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolan-2-yl)methyl carbamate

Chemical formula

[0198] ((2R,3S,5R)-5-(6-Amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolan-2-yl)methyl methylcarbamate was prepared using the same procedure as ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolan-2-yl)methyl isopropylcarbamate, except that propan-2-amine was replaced with methylamine. LCMS(ESI) m / z, C 14 H 15 FN6O4: calculated value 350.1, measured value (M+H) + : 351.2. 1 1H NMR (400 MHz, DMSO-d6) δ (ppm) 8.26 (s, 1H), 7.87 (br s, 2H), 7.13 (br d, J = 4.4 Hz, 1H), 6.24 (dd, J = 7.6, 5.0 Hz, 1H), 5.76 (br d, J = 4.4 Hz, 1H), 4.55 (br d, J = 5.2 Hz, 1H), 4.35 (d, J = 11.6 Hz, 1H), 4.03 (d, J = 11.6 Hz, 1H), 3.60 (s, 1H), 2.70 - 2.79 (m, 1H), 2.54 (s, 3H), 2.40 - 2.45 (m, 1H). 19 19F NMR (376 MHz, DMSO-d6) δ (ppm) -51.75 (s).

[0199] Example 3: Isopropyl (9-((2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2-fluoro-9H-purin-6-yl)carbamate

Chem.

Chem.

[0200] To a mixture of [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-2-ethynyl-3-(4-methylbenzoyl)oxy-tetrahydrofuran-2-yl]methyl 4-methylbenzoate (50 mg, 0.094 mmol, 1 equiv) and Et3N (10 mg, 0.094 mmol, 1 equiv) in DCM (1 mL) at 0 °C was added isopropyl carbonochloridate (23 mg, 0.19 mmol, 2 equiv), and the mixture was stirred at 15 °C for 16 h. The mixture was concentrated under reduced pressure, 2 mL of water was added, and the mixture was extracted with EtOAc (10 mL × 2). The combined organic layers were washed with brine (20 mL) and concentrated under reduced pressure to give (2R,3S,5R)-5-(6-(bis(isopropoxycarbonyl)amino)-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(((4-methylbenzoyl)oxy)methyl)tetrahydrofuran-3-yl 4-methylbenzoate as a crude product, which was used in the next reaction without further purification. LCMS (ESI) m / z, C 36 H 36 FN5O9: Calcd 701.3, Found (M+H) + : 702.1.

[0201] Preparation of Isopropyl (9-((2R,4S,5R)-5-Ethynyl-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2-fluoro-9H-purin-6-yl)carbamate [Chemical Formula]

[0202] To (2R,3S,5R)-5-[6-[Bis(isopropoxycarbonyl)amino]-2-fluoro-9H-purin-9-yl]-2-ethynyl-2-(((4-methylbenzoyl)oxy)methyl)tetrahydrofuran-3-yl] 4-methylbenzoate (66 mg, 0.094 mmol) in THF (1 mL) at -20 °C was added NaOMe (34 mg, 0.19 mmol, 30%, 2 equivalents), and the resulting mixture was stirred at -20 °C for 16 h. Additional NaOMe (17 mg, 0.095 mmol, 30%, 1 equivalent) was added and the mixture was stirred at -20 °C for an additional 40 h. The mixture was neutralized with AcOH (0.1 mL), concentrated under reduced pressure, and purified by flash silica gel chromatography (ISCO®; 4 g SepaFlash® silica flash column, eluting at 20 mL / min with a solvent gradient of 0 - 8% MeOH / DCM) and again by preparative HPLC (column: Waters Xbridge Prep OBD C18 150×30 5u; mobile phase: [water (10 mM NH4HCO3)-ACN]; B%: 5% - 30%, 7 min) to give isopropyl (9-((2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2-fluoro-9H-purin-6-yl)carbamate (3.5 mg, 11% yield) as a white solid. LCMS (ESI) m / z, C 16 H 18 FN5O9: Calculated 379.1, found (M+Na) + : 402.1). 11H NMR (400 MHz, CDCl3) δ (ppm) 8.17 (s, 1H), 7.98 (s, 2H), 6.41 (dd, J =8.8, 5.6 Hz, 1H), 5.13 (dt, J =12.4, 6.4 Hz, 1H), 5.04 (dd, J =11.0, 3.0 Hz, 1H), 4.70 - 4.75 (m, 1H), 4.09 (dd, J =12.4, 2.4 Hz, 1H), 3.84 - 3.93 (m, 1H), 3.06 - 3.15 (m, 1H), 2.83 (s, 1H), 2.48 - 2.56 (m, 2H), 1.36 (d, J =6.0 Hz, 6H). 19 19F NMR (376 MHz, CDCl3) δ (ppm) -46.89 (s).

[0203] Example 4 (Method 1): ((2R,3S,5R)-5-(6-Amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolane-2-yl)methyl ((5-methyl-2-oxo-1,3-dioxol-4-yl)methyl) carbonate

Chemical formula

Chemical formula

[0204] To a mixture of ethanethiol (16 g, 257.5 mmol, 19.1 mL, 1 equiv) and triethylamine (26.1 g, 257.5 mmol, 35.8 mL, 1 equiv) in THF (1 L) was added bis(trichloromethyl) carbonate (76.4 g, 257.5 mmol, 1 equiv) in THF (50 mL) at -15 °C. The mixture was warmed to 18 °C and stirred at 18 °C for 2 h. The mixture was filtered and the filtrate was concentrated under vacuum to give S-ethyl chloromethanethioate (13 g, crude) as a yellow oil, which was used directly in the next reaction without further purification.

[0205] To a mixture of 4-(hydroxymethyl)-5-methyl-1,3-dioxol-2-one (13 g, 99.9 mmol, 1 equiv) in Et2O (800 mL) at 0 °C was added pyridine (7.90 g, 99.9 mmol, 8.1 mL, 1 equiv) and S-ethyl chloromethanethioate (12.45 g, 99.9 mmol, 1.0 equiv) in Et2O (200 mL). The mixture was stirred at 0 °C for 1 h, warmed to 18 °C and stirred at 18 °C for 16 h. The mixture was filtered and concentrated under vacuum, then taken up in DCM (150 mL) and washed with saturated aqueous NaHCO3 (150 mL × 2) and water (150 mL × 2). The mixture was concentrated under reduced pressure and purified by flash silica gel chromatography (ISCO®; 120 g SepaFlash® silica flash column, eluting with a gradient of 0 - 15% ethyl acetate / petroleum ether at 70 mL / min) to give (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl ethylsulfanylformate (9.2 g, 42.2% yield) as a pale yellow oil. 1 H NMR (400 MHz, CDCl3) δ 4.95 (s, 2H), 2.89 (q, J =7.2 Hz, 2H), 2.19 (s, 3H), 1.32 (t, J =7.2 Hz, 3H).

[0206] Preparation of (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl carbonochloridate

Chemical formula

[0207] To a mixture of (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl ethylsulfanylformate (500 mg, 2.29 mmol, 1 equiv) in DCM (50 mL) was added sulfuryl chloride (618.5 mg, 4.58 mmol, 0.46 mL, 2 equiv). The resulting mixture was stirred at 20 °C for 1 h. The reaction mixture was washed with water (50 mL × 2), 5% aqueous Na2CO3 solution (50 mL × 2), brine (50 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl carbonochloridate (350 mg, crude) as a yellow oil. The product was dissolved in 10 mL of DCM and stored in the refrigerator.

[0208] Preparation of tert-butyl (9-((2R,4S,5R)-5-ethynyl-4-hydroxy-5-(((((5-methyl-2-oxo-1,3-dioxol-4-yl)methoxy)carbonyl)oxy)methyl)tetrahydrofuran-2-yl)-2-fluoro-9H-purin-6-yl)carbamate

Chemical formula

[0209] To a mixture of tert-butyl N-[9-[(2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]-2-fluoro-purin-6-yl]carbamate (100 mg, 0.25 nmol, 1 equiv) in pyridine (2 mL) was added (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl carbonochloridate (140 mg, 0.73 mmol, 4 mL, 2.86 equiv, 35 mg / mL in DCM) at 20 °C, and the mixture was stirred at 20 °C for 16 h. The mixture was concentrated under reduced pressure and purified by flash silica gel chromatography (ISCO®; 4 g SepaFlash® silica flash column, eluting at 25 mL / min with a 0 - 5% MeOH / DCM gradient) to give [(2R,3S,5R)-5-[6-(tert-butoxycarbonylamino)-2-fluoro-purin-9-yl]-2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl]methyl (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl carbonate (45 mg, 32.2% yield) as a yellow solid. LCMS (ESI) m / z, C 23 H 24 FN5O 10 : calculated 549.2, found (M + H) + : 550.1.

[0210] Preparation of ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl)methyl ((5-methyl-2-oxo-1,3-dioxol-4-yl)methyl) carbonate

Chemical formula

[0211] A mixture of [(2R,3S,5R)-5-[6-(tert-Butoxycarbonylamino)-2-fluoro-purin-9-yl]-2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl]methyl (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl carbonate (45 mg, 0.082 mmol, 1 equiv) in toluene (1 mL) was added with TFA (154 mg, 1.35 mmol, 0.1 mL, 16.5 equiv) at 20 °C. The mixture was stirred at 20 °C for 16 h and then concentrated under reduced pressure. The obtained residue was purified by preparative HPLC (column: Boston Green ODS 150×30 mm×5 um; mobile phase: [water (0.2% FA)-ACN], B%: 13% - 43%, 8 min) to give ((2R,3S,5R)-5-(6-Amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl)methyl ((5-methyl-2-oxo-1,3-dioxol-4-yl)methyl) carbonate (11 mg, yield 25.4%) as a white solid. LCMS (ESI) m / z, C 18 H 16 FN5O6: calculated value 449.1, measured value (M+H) + : 450.1. 19 F NMR (376 MHz, CD3OD) δ (ppm) -53.00 (s). 1 H NMR (400 MHz, CD3CN) 7.92 (s, 1H), 6.41 - 6.21 (m, 3H), 4.87 (d, J = 5.2 Hz, 2H), 4.77 - 4.67 (m, 1H), 4.51 (d, J = 11.6 Hz, 1H), 4.30 (d, J = 11.6 Hz, 1H), 3.75 (d, J = 6.4 Hz, 1H), 3.00 (s, 1H), 2.89 - 2.81 (m, 1H), 2.61 - 2.52 (m, 1H), 2.10 (s, 3H).

[0212] Example 4 (Method 2): ((2R,3S,5R)-5-(6-Amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolane-2-yl)methyl ((5-methyl-2-oxo-1,3-dioxol-4-yl)methyl) carbonate

Chem.

[0213] ((2R,3S,5R)-5-(6-Amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolane-2-yl)methyl (4-nitrophenyl) carbonate Preparation

Chem.

[0214] To a mixture of (2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)oxolan-3-ol (100 mg, 0.34 mmol, 1 equiv) in pyridine (5 mL) was added (4-nitrophenyl) carbonochloridate (82 mg, 0.41 mmol, 1.2 equiv), and the mixture was stirred at 26 °C for 16 h. (4-Nitrophenyl) carbonochloridate (82 mg, 0.41 mmol, 1.2 equiv) was added, and the mixture was stirred at 26 °C for 24 h. The reaction solution was purified by flash silica gel chromatography (ISCO (registered trademark); 4 g of SepaFlash (registered trademark) silica flash column, eluent with a 0 - 5% DCM / MeOH gradient at 25 mL / min), and [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-2-ethynyl-3-hydroxy-oxolan-2-yl]methyl (4-nitrophenyl) carbonate (80 mg, yield 51.2%) was obtained as a white solid. LCMS (ESI) m / z, C 19 H 15 FN6O7: Calculated value 458.4, measured value (M + H) + : 459.1。

[0215] Preparation of ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolane-2-yl)methyl ((5-methyl-2-oxo-1,3-dioxol-4-yl)methyl) carbonate

Chemical formula

[0216] To a mixture of [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-2-ethynyl-3-hydroxy-oxolane-2-yl]methyl (4-nitrophenyl) carbonate (150 mg, 0.298 mmol, 1 equiv) and 4-(hydroxymethyl)-5-methyl-1,3-dioxol-2-one (96 mg, 0.745 mmol, 2.5 equiv) in THF (3 mL) was added DMAP (3.6 mg, 0.023 mmol, 0.1 equiv), and the mixture was stirred at 25 °C for 2 h. The reaction mixture was purified by flash silica gel chromatography (ISCO®; 24 g SepaFlash® silica flash column, eluting with a gradient of 0 - 2.5% DCM / MeOH at 25 mL / min) to give [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-2-ethynyl-3-hydroxy-oxolane-2-yl]methyl (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl carbonate (85 mg, 63.5% yield) as a white solid. LCMS (ESI) m / z, C 18 H 16 Calculated for FN5O8 449.4, found 450.1 (M+H) + . 11H NMR (400 MHz, CD3CN) δ (ppm) 7.91 (s, 1H), 6.42 - 6.16 (m, 3H), 4.93 - 4.79 (m, 2H), 4.76 - 4.67 (m, 1H), 4.53 - 4.46 (m, 1H), 4.34 - 4.25 (m, 1H), 3.77 - 3.69 (m, 1H), 3.00 (s, 1H), 2.90 - 2.78 (m, 1H), 2.62 - 2.50 (m, 1H), 2.10 (s, 3H). 19 19F NMR (376 MHz, CD3CN) δ (ppm) -52.87 (s, 1F).

[0217] Example 4 (Method 3): ((2R,3S,5R)-5-(6-Amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolane-2-yl)methyl ((5-methyl-2-oxo-1,3-dioxol-4-yl)methyl) carbonate

Chem.

[0218] Preparation of ((2R,3S,5R)-5-(6-Amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolane-2-yl)methyl (4-nitrophenyl) carbonate

Chem.

[0219] To a mixture of (2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (5 g, 17.05 mmol, 1 equiv) in pyridine (50 mL) was added dropwise (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl carbonochloridate (16.42 g, 85.25 mmol, 5 equiv) in DCM (16 mL) at 0 °C over 2 h, and then the mixture was stirred at 16 °C for 10 min. The mixture was diluted with DCM (200 mL), washed with water (150 mL), brine (150 mL×2), dried over Na2SO4, filtered, and concentrated. The obtained residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash® silica flash column, eluting with a 0 - 5% MeOH / DCM gradient at 65 mL / min) to give [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl]methyl (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl carbonate (5.10 g, 11.35 mmol, 66.6% yield) as a pale yellow solid. LCMS (ESI) m / z, C 18 H 16 FN5O8: calculated 449.4, found (M+H) + : 450.1. 1 H NMR (400 MHz, CD3CN) δ (ppm) 7.92 (s, 1H), 6.34 (br s, 2H), 6.29 - 6.23 (m, 1H), 4.93 - 4.81 (m, 2H), 4.77 - 4.69 (m, 1H), 4.51 (d, J = 11.6 Hz, 1H), 4.30 (d, J = 11.6 Hz, 1H), 3.74 (d, J = 6.4 Hz, 1H), 3.00 (s, 1H), 2.91 - 2.79 (m, 1H), 2.62 - 2.51 (m, 1H), 2.10 (s, 3H). 19 F NMR (376 MHz, CD3CN) δ (ppm) -52.84 (s, 1F).

[0220] Recrystallization of ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolane-2-yl)methyl (4-nitrophenyl) carbonate

Chem.

[0221] A mixture of [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-2-ethynyl-3-hydroxy-oxolane-2-yl]methyl (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl carbonate (9.5 g, 21.14 mmol, 1 equiv) in MeCN (50 mL) and EtOAc (50 mL) was heated at 80 o for 30 min and the dissolution of the solid was observed. After cooling to room temperature (20 °C), the mixture was stirred at 20 °C for 16 h. The mixture was filtered and the filter cake was dried in vacuo to give [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-2-ethynyl-3-hydroxy-oxolane-2-yl]methyl (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl carbonate (8.0 g, 17.80 mmol, 84.2% yield) as a white solid. LCMS (ESI) m / z, C 18 H 16 FN5O8: calcd 449.4, found (M+H) + : 450.1. 1 H NMR (400 MHz, CD3CN) δ (ppm) 7.92 (s, 1H), 6.31 (br s, 2H), 6.27 - 6.24 (m, 1H), 4.92 - 4.81 (m, 2H), 4.77 - 4.69 (m, 1H), 4.51 (d, J = 11.6 Hz, 1H), 4.30 (d, J = 11.6 Hz, 1H), 3.73 (d, J = 6.4 Hz, 1H), 3.00 (s, 1H), 2.89 - 2.81 (m, 1H), 2.62 - 2.51 (m, 1H), 2.10 (s, 3H). 1919F NMR (376 MHz, CD3CN) δ (ppm) -52.84 (s, 1F).

[0222] Example 5 (Method 1): 4-(((9-((2R,4S,5R)-5-Ethynyl-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2-fluoro-9H-purin-6-yl)amino)methyl)-5-methyl-1,3-dioxol-2-one

Chemical Structure

Chemical Structure

[0223] To a mixture of tert-butyl N-[9-[(2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]-2-fluoro-purin-6-yl]carbamate (100 mg, 0.25 nmol, 1 equiv) in DMF (5 mL) were added K2CO3 (70 mg, 0.51 mmol, 2 equiv) and 4-(bromomethyl)-5-methyl-1,3-dioxol-2-one (147 mg, 0.76 mmol, 3 equiv). The reaction mixture was then heated at 60 °C for 12 h, concentrated, and then diluted with H2O (30 mL). The resulting mixture was extracted with EtOAc (30 × 3 mL). The combined organic layers were washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated. The resulting residue was purified by flash silica gel chromatography (ISCO®; 4 g SepaFlash® silica flash column, eluting at 30 mL / min with a 0 - 10% methanol / dichloromethane gradient) to give tert-butyl N-[9-[(2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]-2-fluoro-purin-6-yl]-N-[(5-methyl-2-oxo-1,3-dioxol-4-yl)methyl]carbamate (45 mg, 35.0% yield) as a pale yellow oil. 1 H NMR (400 MHz, CDCl3) δ (ppm) 8.02 (s, 1H), 6.40 - 6.43 (m, 1H), 5.02 (s, 2H), 4.70 - 4.73 (m, 2H), 4.09 (d, J =12 Hz, 1H), 3.90 - 3.92 (m, 1H), 3.11 - 3.14 (m, 1H), 2.85 (s, 1H), 2.50 - 2.55 (m, 1H), 2.45 (bs, 1H), 2.22 (s, 3H), 1.53(s, 9H).

[0224] Preparation of 4-(((9-((2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2-fluoro-9H-purin-6-yl)amino)methyl)-5-methyl-1,3-dioxol-2-one

Chem.

[0225] To a solution of tert-butyl N-[9-[(2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]-2-fluoro-purin-6-yl]-N-[(5-methyl-2-oxo-1,3-dioxol-4-yl)methyl]carbamate (40 mg, 0.079 mmol, 1 equiv) in dichloromethane (DCM) (3 mL) was added TFA (0.5 mL) at 25 °C. The mixture was stirred at 25 °C for 12 h. The reaction mixture was concentrated and purified by preparative HPLC (column: Boston Green ODS 150*30 mm*5um; mobile phase: [water (0.2% FA)-ACN], B%: 15%-45%, 8 min) to give 4-(((9-((2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2-fluoro-9H-purin-6-yl)amino)methyl)-5-methyl-1,3-dioxol-2-one (1.7 mg, 5% yield) as a white solid. LCMS (ESI) m / z, C 17 H 16 FN5O6: calculated value 405.1, measured value (M+H) + : 406.1. (M+Na) + : 428.1. 1 H NMR (400 MHz, CD3OD) δ (ppm) 8.26 (s, 1H), 6.38 - 6.35 (m, 1H), 4.76 - 4.72 (m, 1H), 4.51 (s, 1H), 3.87 - 3.84 (d, J = 12 Hz, 1H), 3.78 - 3.75 (d, J = 12 Hz, 1H), 3.09 (s, 2H), 2.80 - 2.75 (m, 1H), 2.64 - 2.57 (m, 1H), 2.24 (s, 3H). 19 F NMR (376 MHz, CD3OD) δ (ppm) -53.00.

[0226] Example 5 (Method 2): 4-(((9-((2R,4S,5R)-5-Ethynyl-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2-fluoro-9H-purin-6-yl)amino)methyl)-5-methyl-1,3-dioxol-2-one

Chem.

[0227] Preparation of tert-butyl-N-[9-[(2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]-2-fluoro-purin-6-yl]-N-[(5-methyl-2-oxo-1,3-dioxol-4-yl)methyl]carbamate

Chem.

[0228] To a solution of tert-butyl N-[9-[(2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]-2-fluoro-purin-6-yl]carbamate (300 mg, 0.76 mmol, 1 equiv) in DMF (5 mL) was added NaHCO3 (128 mg, 1.52 mmol, 2 equiv), followed by the addition of 4-(bromomethyl)-5-methyl-1,3-dioxol-2-one (294 mg, 1.52 mmol, 2 equiv). The mixture was stirred at 25 °C for 48 h. The mixture was concentrated. The resulting residue was purified by flash silica gel chromatography (ISCO (registered trademark); 4 g SepaFlash (registered trademark) silica flash column, eluting with a 0–100% ethyl acetate / petroleum ether gradient at 20 mL / min) to give tert-butyl N-[9-[(2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]-2-fluoro-purin-6-yl]-N-[(5-methyl-2-oxo-1,3-dioxol-4-yl)methyl]carbamate (200 mg, 52% yield) as a white solid. 11H NMR (400 MHz, CDCl3) δ (ppm) 7.98 (s, 1H), 6.45 - 6.33 (m, 1H), 4.99 (s, 1H), 5.03 - 4.94 (m, 1H), 4.78 (br d, J = 11.2 Hz, 1H), 4.71 (br s, 1H), 4.16 - 4.02 (m, 2H), 3.11 (br s, 1H), 2.80 (s, 1H), 2.49 (br d, J = 7.2 Hz, 1H), 2.42 (br s, 1H), 2.19 (s, 3H), 1.55 (s, 9H).

[0229] Preparation of 4 - [[[9 - [(2R,4S,5R) - 5 - ethynyl - 4 - hydroxy - 5 - (hydroxymethyl)tetrahydrofuran - 2 - yl] - 2 - fluoro - purin - 6 - yl]amino]methyl] - 5 - methyl - 1,3 - dioxol - 2 - one [Chemical formula]

[0230] To a solution of tert - butyl N - [9 - [(2R,4S,5R) - 5 - ethynyl - 4 - hydroxy - 5 - (hydroxymethyl)tetrahydrofuran - 2 - yl] - 2 - fluoro - purin - 6 - yl] - N - [(5 - methyl - 2 - oxo - 1,3 - dioxol - 4 - yl)methyl]carbamate (50 mg, 0.10 mmol, 1 equivalent) in CH3CN (5 mL) was added ZnBr2 (45 mg, 0.20 mmol, 2 equivalents). The mixture was stirred at 25 °C for 48 h. The reaction mixture was filtered and the filtrate was concentrated. The resulting residue was purified by preparative HPLC (FA conditions; column: 3_Phenomenex Luna C18 75×30 mm×3 um; mobile phase: [water (0.2% FA) - ACN], B%: 22% - 52%, 6 min) to give 4 - [[[9 - [(2R,4S,5R) - 5 - ethynyl - 4 - hydroxy - 5 - (hydroxymethyl)tetrahydrofuran - 2 - yl] - 2 - fluoro - purin - 6 - yl]amino]methyl] - 5 - methyl - 1,3 - dioxol - 2 - one (19.8 mg, 48.8% yield) as a white solid. LCMS (ESI) m / z, C17 H 16 FN5O6: Calculated value 405.34, measured value (M+H) + : 406.1 1 1H NMR (400 MHz, CD3OD) δ (ppm) 8.26 (s, 1H), 6.38 - 6.35 (m, 1H), 4.76 - 4.72 (m, 1H), 4.51 (br s, 2H), 3.87 - 3.84 (d, J = 12 Hz, 1H), 3.78 - 3.75 (d, J = 12 Hz, 1H), 3.09 (s, 1H), 2.80 - 2.75 (m, 1H), 2.64 - 2.57 (m, 1H), 2.24 (s, 3H). 19 19F NMR (376 MHz, CD3OD) δ (ppm) -52.33

[0231] Example 6 ((2R,3S,5R)-5-(6-Amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolane-2-yl)methyl (2-(methyl((5-methyl-2-oxo-1,3-dioxol-4-yl)methyl)amino)ethyl)carbamate

Chemical Structure

Chemical Structure

[0232] To a mixture of tert-butyl N-[2-(methylamino)ethyl]carbamate (451 mg, 2.59 mmol, 1 equiv) and K2CO3 (393.9 mg, 2.85 mmol, 1.1 equiv) in DMF (15 mL) was added 4-(bromomethyl)-5-methyl-1,3-dioxol-2-one (500 mg, 2.59 mmol, 1 equiv), and the mixture was stirred at 20 °C for 16 h. The mixture was concentrated under reduced pressure and purified by flash silica gel chromatography (ISCO®; 12 g SepaFlash® silica flash column, eluting with a 0–20% ethyl acetate / petroleum ether gradient at 30 mL / min) to give tert-butyl N-[2-[methyl-[(5-methyl-2-oxo-1,3-dioxol-4-yl)methyl]amino]ethyl]carbamate (500 mg, 1.75 mmol, 67.4% yield) as a yellow oil. 1 H NMR (400 MHz, DMSO-d6) δ (ppm) 6.65 (m, 1H), 3.37 (s, 2H), 2.99 (m, 2H), 2.39 (m, 2H), 2.18 (s, 3H), 2.09 (s, 3H), 1.37 (s, 9H).

[0233] Preparation of 4-(((2-aminoethyl)(methyl)amino)methyl)-5-methyl-1,3-dioxol-2-one

Chemical formula

[0234] To a mixture of tert-butyl N-[2-[methyl-[(5-methyl-2-oxo-1,3-dioxol-4-yl)methyl]amino]ethyl]carbamate (200 mg, 0.699 nmol, 1 equiv) in dichloromethane (2 mL) at 20 °C was added TFA (1.54 g, 13.51 mmol, 1 mL, 19.34 equiv), and the mixture was stirred at 20 °C for 16 h. The mixture was concentrated under reduced pressure to give 4-[[2-aminoethyl(methyl)amino]methyl]-5-methyl-1,3-dioxol-2-one (120 mg, crude, 3TFA) as a yellow oil. LCMS (ESI) m / z, C8H14 N2O3: Calculated value 186.1, measured value (M+H) + : 187.2

[0235] Preparation of tert-butyl (9-((2R,4S,5R)-5-ethynyl-4-hydroxy-5-(((2-(methyl((5-methyl-2-oxo-1,3-dioxol-4-yl)methyl)amino)ethyl)carbamoyl)oxy)methyl)tetrahydrofuran-2-yl)-2-fluoro-9H-purin-6-yl)carbamate [Chemical formula]

[0236] To a mixture of [(2R,3S,5R)-5-[6-(tert-butoxycarbonylamino)-2-fluoro-purin-9-yl]-2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl]methyl (4-nitrophenyl) carbonate (34 mg, 0.060 mmol, 1 equivalent) and triethylamine (30 mg, 0.30 mmol, 5 equivalents) in dichloromethane (2 mL) at 20 °C was added 4-[[2-aminoethyl(methyl)amino]-5-methyl-1,3-dioxol-2-one (38 mg, 0.072 mmol, 1.2 equivalents, 3TFA), and the mixture was stirred at 20 °C for 16 hours. Then, the mixture was concentrated under reduced pressure and purified by flash silica gel chromatography (ISCO (registered trademark); 4 g of SepaFlash (registered trademark) silica flash column, eluent with a 0 - 5% dichloromethane / MeOH gradient at 25 mL / min) to obtain tert-butyl N-[9-[(2R,4S,5R)-5-ethynyl-4-hydroxy-5-[2-[methyl-[(5-methyl-2-oxo-1,3-dioxol-4-yl)methyl]amino]ethylcarbamoyloxy)methyl]tetrahydrofuran-2-yl]-2-fluoro-purin-6-yl]carbamate (34 mg, yield 92.1%) as a yellow solid. LCMS(ESI) m / z, C 26 H 32 FN7O9: Calculated value 605.2, measured value (M+Na) + : 628.2

[0237] Preparation of ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolane-2-yl)methyl (2-(methyl((5-methyl-2-oxo-1,3-dioxol-4-yl)methyl)amino)ethyl)carbamate

Chem.

[0238] To a mixture of tert-butyl N-[9-[(2R,4S,5R)-5-ethynyl-4-hydroxy-5-[2-[methyl-[(5-methyl-2-oxo-1,3-dioxol-4-yl)methyl]amino]ethylcarbamoyloxymethyl]tetrahydrofuran-2-yl]-2-fluoro-purin-6-yl]carbamate (34 mg, 0.055 mmol, 1 equiv) in toluene (1 mL) was added TFA (129 mg, 1.13 mmol, 0.084 mL, 20.45 equiv), and the mixture was stirred at 20 °C for 16 h. The mixture was concentrated under reduced pressure and purified by preparative HPLC (column: Phenomenex Gemini-NX 150×30 mm×5 μm; mobile phase: [water (0.05% HCl)-ACN], B%: 0% - 30%, 7 min), then re-purified by preparative HPLC (column: Welch Xtimate C18 150×25 mm×5 μm; mobile phase: [water (0.2% FA)-ACN], B%: 1% - 20%, 8 min) to give 4-(((9-((2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2-fluoro-9H-purin-6-yl)amino)methyl)-5-methyl-1,3-dioxol-2-one (2.0 mg, yield 20.0%) as a white solid. LCMS (ESI) m / z, C 21 H 24 FN7O7: calculated 505.2, found (M+H) + : 506.3, (M+Na) + : 528.2. 11H NMR (400 MHz, CD3OD) δ (ppm) 8.15 (s, 1H), 6.32 (dd, J = 8.0, 4.0 Hz, 1H), 4.80 (m, 1H), 4.47 (d, J = 11.6 Hz, 1H), 4.24 (d, J = 11.6 Hz, 1H), 3.45 (s, 2H), 3.19 (t, J = 6.8, 2H), 3.17 (s, 1H), 2.84 (m, 1H), 2.66 (m, 1H), 2.51 (t, J = 6.8, 2H), 2.30 (s, 3H), 2.14 (s, 3H). 19 19F NMR (376 MHz, CD3OD) δ (ppm) -52.93 (s).

[0239] Example 7: ((2R,3S,5R)-5-(6-Amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolane-2-yl)methyl 4-(methyl(((5-methyl-2-oxo-1,3-dioxolan-4-yl)methoxy)carbonyl)amino)butanoate

Chemical Structure

Chemical Structure

[0240] A mixture of 1-methylpyrrolidin-2-one (5.0 g, 50.44 mmol, 4.90 mL, 1 equiv), HCl (51.0 g, 531.53 mmol, 50 mL, 38% purity, 10.54 equiv) was stirred at 165 °C for 16 h. The reaction mixture was concentrated. The resulting residue was triturated from acetone to give 4-(methylamino)butanoic acid (4 g, 51.6% yield, HCl salt) as a white solid.

[0241] Preparation of 4-[methyl-[(5-methyl-2-oxo-1,3-dioxolan-4-yl)methoxycarbonyl]amino]butanoic acid [Chemistry]

[0242] To a solution of 4-(methylamino)butanoic acid HCl salt (1.36 g, 8.83 mmol, 1 equiv) in H2O (25 mL) was added NaHCO3 (2.23 g, 26.49 mmol, 1.03 mL, 3 equiv), followed by (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl carbonochloridate (1.7 g, 8.83 mmol, 1 equiv) in THF (25 mL). The mixture was stirred at 20 °C for 16 h. The reaction mixture was concentrated and purified by flash silica gel chromatography (ISCO (registered trademark); 12 g SepaFlash (registered trademark) silica flash column, eluting with 0 - 5% methanol / dichloromethane at 20 mL / min) to give 4-[methyl-[(5-methyl-2-oxo-1,3-dioxol-4-yl)methoxycarbonyl]amino]butanoic acid (1.5 g, 62.2% yield) as a colorless oil. 1 1H NMR (400 MHz, DMSO-d6) δ (ppm) 12.07 (br s, 1H), 4.89 (s, 2H), 3.22 (br t, J = 7.2 Hz, 2H), 2.81 (s, 3H), 2.28 - 2.03 (m, 5H), 1.69 (m, 2H).

[0243] Preparation of (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl N-(4-chloro-4-oxo-butyl)-N-methyl-carbamate [Chemistry]

[0244] To a solution of 4-[methyl-[(5-methyl-2-oxo-1,3-dioxol-4-yl)methoxycarbonyl]amino]butanoic acid (70 mg, 0.256 mmol, 1 equiv) in DCM (5 mL) was added DMF (0.2 mg, 0.0026 mmol, 0.01 equiv) and oxalyl chloride (65 mg, 0.512 mmol, 0.045 mL, 2 equiv) at 0 °C. The mixture was stirred at 20 °C for 0.5 h. The reaction was concentrated to dryness to afford (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl N-(4-chloro-4-oxo-butyl)-N-methyl-carbamate (50 mg, crude) as a yellow oil, which was used in the next reaction without further purification.

[0245] Preparation of [(2R,3S,5R)-5-[6-(tert-butoxycarbonylamino)-2-fluoro-purin-9-yl]-2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl]methyl 4-[methyl-[(5-methyl-2-oxo-1,3-dioxol-4-yl)methoxycarbonyl]amino]butanoate

Chemical formula

[0246] A solution of tert-butyl N-[9-[(2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]-2-fluoro-purin-6-yl]carbamate (20 mg, 0.0508 mmol, 1 equiv) in pyridine (2 mL) was stirred at 20 °C for 16 h after addition of (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl-(4-chloro-4-oxo-butyl)-N-methyl-carbamate (66 mg, 0.226 mmol, 4.45 equiv). The reaction mixture was concentrated to give [(2R,3S,5R)-5-[6-(tert-butoxycarbonylamino)-2-fluoro-purin-9-yl]-2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl]methyl 4-[methyl-[(5-methyl-2-oxo-1,3-dioxol-4-yl)methoxycarbonyl]amino]butanoate (35 mg, crude) as a yellow oil, which was used in the next reaction without further purification.

[0247] Preparation of ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl)methyl 4-(methyl(((5-methyl-2-oxo-1,3-dioxol-4-yl)methoxy)carbonyl)amino)butanoate

Chemical formula

[0248] A solution of [(2R,3S,5R)-5-[6-(tert-butoxycarbonylamino)-2-fluoro-purin-9-yl]-2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl]methyl 4-[methyl-[(5-methyl-2-oxo-1,3-dioxol-4-yl)methoxycarbonyl]amino]butanoate (25 mg, 0.0386 mmol, 1 equiv) in toluene (2 mL) was added with TFA (770 mg, 0.675 mmol, 0.50 mL, 10%, 17.52 equiv). The mixture was stirred at 20 °C for 16 h and then concentrated to dryness. The residue was purified by preparative HPLC (TFA conditions; column: Boston Green ODS 150*30mm*5um; mobile phase: [water (0.075% TFA)-ACN], B%: 20% - 50%, 12 min) to give (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl)methyl 4-(methyl(((5-methyl-2-oxo-1,3-dioxol-4-yl)methoxy)carbonyl)amino)butanoate (1.8 mg, yield 8.51%) as a pale yellow solid. LCMS (ESI) m / z, C 23 H 26 FN6O 9、 Calculated 548.2, found (M+H) + : 549.2. 1 H NMR (400 MHz, CD3OD) δ (ppm) 8.14 (s, 1H), 6.29 (m, 1H), 4.88 (s, 2H), 4.44 (t, J = 11.2 Hz, 1H), 4.25 (t, J = 11.2 Hz, 1H), 3.26 - 3.13 (m, 3H), 2.96 - 2.86 (m, 2H), 2.82 (s, 2H), 2.64 (m, 1H), 2.25 (m, 2H), 2.16 - 2.07 (m, 3H), 1.74 (m, 2H), 1.31 (s, 1H). 19 F NMR (376 MHz, CD3OD) δ (ppm) -52.82 (s).

[0249] Example 8: N-(9-((2R,4S,5R)-5-Ethynyl-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2-fluoro-9H-purin-6-yl)butylamide

Chem.

[0250] Preparation of [(2R,3S,5R)-5-[6-[Butanoyl(tert-butoxycarbonyl)amino]-2-fluoro-purin-9-yl]-2-ethynyl-3-(4-methylbenzoyl)oxy-tetrahydrofuran-2-yl]methyl 4-methylbenzoate

Chem.

[0251] A solution of [(2R,3S,5R)-5-[6-(tert-butoxycarbonylamino)-2-fluoro-purin-9-yl]-2-ethynyl-3-(4-methylbenzoyl)oxy-tetrahydrofuran-2-yl]methyl 4-methylbenzoate (74 mg, 0.118 mmol, 1 equiv) in DCM (5 mL) was added with DMAP (1.5 mg, 0.012 mmol, 0.1 equiv), TEA (36 mg, 0.354 mmol, 3.0 equiv) and butanoyl chloride (25 mg, 0.235 mmol, 2 equiv). The mixture was stirred at 25 °C for 16 h. The mixture was concentrated, diluted with H2O (30 mL), and then extracted with EtOAc (15 × 3 mL). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated. The resulting residue was purified by flash silica gel chromatography (ISCO®; 4 g SepaFlash® silica flash column, eluting at 18 mL / min with a gradient of 0 - 20% ethyl acetate / petroleum ether) to give [(2R,3S,5R)-5-[6-[butanoyl(tert-butoxycarbonyl)amino]-2-fluoro-purin-9-yl]-2-ethynyl-3-(4-methylbenzoyl)oxy-tetrahydrofuran-2-yl]methyl 4-methylbenzoate (30 mg, yield 30%) as a pale yellow oil. LCMS (ESI) m / z, C 37 H 38 FN5O8: calcd 700.3, found (M+H) + : 701.3.

[0252] Preparation of [(2R,3S,5R)-5-[6-(butanoylamino)-2-fluoro-purin-9-yl]-2-ethynyl-3-(4-methylbenzoyl)oxy-tetrahydrofuran-2-yl]methyl 4-methylbenzoate

Chem.

[0253] A solution of [(2R,3S,5R)-5-[6-[butanoyl(tert-butoxycarbonyl)amino]-2-fluoro-purin-9-yl]-2-ethynyl-3-(4-methylbenzoyl)oxy-tetrahydrofuran-2-yl]methyl 4-methylbenzoate (30 mg, 0.0428 mmol, 1 equiv) in toluene (1 mL) was added with TFA (0.1 mL). The mixture was stirred at 25 °C for 16 h. The mixture was diluted with H2O (2 mL), extracted with EtOAc (3 × 3 mL), the combined organic layers were washed with brine (15 × 3 mL), dried over Na2SO4, filtered, and concentrated. The obtained residue was purified by preparative TLC (SiO2, petroleum ether / ethyl acetate = 1:1) to give [(2R,3S,5R)-5-[6-(butanoylamino)-2-fluoro-purin-9-yl]-2-ethynyl-3-(4-methylbenzoyl)oxy-tetrahydrofuran-2-yl]methyl 4-methylbenzoate (18 mg, yield 64%) as a pale yellow oil. LCMS (ESI) m / z, C 32 H 30 FN5O6: calculated 600.2, found (M+H) + : 601.3.

[0254] Preparation of N-[9-[(2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]-2-fluoro-purin-6-yl]butanamide

Chem.

[0255] A solution of [(2R,3S,5R)-5-[6-(butanoylamino)-2-fluoro-purin-9-yl]-2-ethynyl-3-(4-methylbenzoyl)oxy-tetrahydrofuran-2-yl]methyl 4-methylbenzoate (18 mg, 0.030 mmol, 1 equiv) in THF (2 mL) was added with CH3ONa (54 mg, 0.30 mmol, 30% purity in MeOH, 10 equiv). The mixture was stirred at -25 °C for 4 h. After acidifying the mixture to pH = 7 with HOAc, the mixture was concentrated. The obtained residue was purified by preparative HPLC (FA conditions; column: 3_Phenomenex Luna C18 75×30 mm×3 um; mobile phase: [water (0.2% FA)-ACN], B%: 15% - 45%, 6 min) to give N-[9-[(2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]-2-fluoro-purin-6-yl]butanamide (4.4 mg, yield 40%) as a white solid. LCMS (ESI) m / z, C 16 H 18 FN5O4: calculated value 364.1, measured value (M+H) + : 365.2. 1 H NMR (400 MHz, CD3CN) δ (ppm) 9.15 (bs, 1H), 8.26 (s, 1H), 6.37 - 6.34 (m, 1H), 4.70 (t, J = 6.4 Hz, 1H), 3.82 - 3.78 (m, 1H), 3.74 - 3.69 (m, 1H), 2.96 - 2.93 (m, 1H), 2.84 - 2.75 (m, 2H), 2.66 (t, J = 7.6 Hz, 2H), 2.59 - 2.52 (m, 2H), 1.74 - 1.67 (m, 2H), 0.99 (t, J = 7.6 Hz, 3H). 19 F NMR (376 MHz, CD3CN) δ (ppm) -51.9 (s, 1F).

[0256] Example 9: [(2R,3S,5R)-5-(6-Amino-2-fluoro-purin-9-yl)-4,4-dideuterio 2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl]methyl (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl carbonate

Chem.

[0257] Preparation of [(2R,3S,5R)-5-(6-Amino-2-fluoro-purin-9-yl)-4,4-dideuterio 2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl]methyl (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl carbonate

Chem.

[0258] To a mixture of (2R,3S,5R)-5-(6-Amino-2-fluoro-purin-9-yl)-4,4-dideuterio 2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (60 mg, 0.203 mmol, 1 equiv) in pyridine (0.5 mL) was added dropwise (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl carbonochloridate (195 mg, 1.02 mmol, 5 equiv) in DCM (0.5 mL). The resulting mixture was stirred at 25 °C for 3 h. The reaction mixture was concentrated. The obtained residue was purified by flash silica gel chromatography (ISCO (registered trademark); 4 g SepaFlash (registered trademark) silica flash column, eluting with a 0 - 5% methanol / dichloromethane gradient at 18 mL / min) to give [(2R,3S,5R)-5-(6-Amino-2-fluoro-purin-9-yl)-4,4-dideuterio 2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl]methyl (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl carbonate (21 mg, 22.9% yield) as a white solid. LCMS (ESI) m / z, C 18 H 14D2FN5O8: Calculated value 451.11, measured value (M+H) + : 452.1 1 H NMR (400 MHz, CD3CN) δ (ppm) 7.92 (s, 1H), 6.31 (s, 2H), 6.25 (s, 1H), 4.9 - 4.82 (m, 2H), 4.73 - 4.71 (d, J = 8 Hz, 1H), 4.52 - 4.50 (d, J = 8 Hz, 1H), 4.32 - 4.29 (d, J = 12 Hz, 1H), 3.74 - 3.72 (d, J = 8 Hz, 1H), 3.00 (s, 1H), 2.10 (s, 3H). 19 F NMR (376 MHz, CD3CN) δ (ppm) -52.84 (s, 1F).

[0259] Example 10: [(2R,3S,5R)-5-(6-Amino-2-fluoro-purin-9-yl)-4,4-dideuterio-2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl]-dideuterio-methyl](5-methyl-2-oxo-1,3-dioxol-4-yl)methyl carbonate

Chemical Structure

[0260] Preparation of [(2R,3S,5R)-5-(6-Amino-2-fluoro-purin-9-yl)-4,4-dideuterio-2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl]-dideuterio-methyl](5-methyl-2-oxo-1,3-dioxol-4-yl)methyl carbonate

Chemical Structure

[0261] To a mixture of (2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-4,4-dideuterio-2-[dideuterio(hydroxy)methyl]-2-ethynyl-tetrahydrofuran-3-ol (50 mg, 0.168 mmol, 1 equiv) in pyridine (0.5 mL) was added dropwise (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl carbonochloridate (162 mg, 0.84 mmol, 5 equiv) in DCM (0.5 mL). The resulting mixture was stirred at 25 °C for 3 h. The reaction mixture was concentrated. The obtained residue was purified by flash silica gel chromatography (ISCO®; 4 g SepaFlash® silica flash column, eluent with 0 - 5% methanol / dichloromethane gradient at 18 mL / min) to give [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-4,4-dideuterio 2-ethynyl-3-hydroxy-tetrahydrofuran-2-yl]-dideuterio methyl](5-methyl-2-oxo-1,3-dioxol-4-yl)methyl carbonate (40.6 mg, 53% yield) as a white solid. LCMS (ESI) m / z, C 18 H 12 D4FN5O8: calculated 453.3, found (M+H) + : 454.2. 1 H NMR (400 MHz, DMSO-d6) δ (ppm) 7.92 (s, 1H), 6.31 (s, 2H), 6.25 (s, 1H), 4.91 - 4.82 (m, 2H), 4.72 - 4.71 (d, J = 4 Hz, 1H), 3.74 - 3.72 (d, J = 8 Hz, 1H), 3.00 (s, 1H), 2.10 (s, 3H). 19 F NMR (376 MHz, DMSO-d6) δ (ppm) -52.85 (s, 1F).

[0262] Example 11: ((2R,3S,5R)-5-(6-Amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxy tetrahydrofuran-2-yl)methyl butyl carbonate [Chemical formula]

[0263] Preparation of ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolan-2-yl)methyl butyl carbonate [Chemical formula]

[0264] To a solution of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)oxolan-3-ol (50 mg, 0.17 mmol, 1 eq) in pyridine (0.5 mL) was slowly added butyl carbonochloridate (26 mg, 0.187 mmol, 0.024 mL, 1.1 eq) at 0 °C, and the mixture was stirred for 2 h. The reaction mixture was concentrated, and the resulting residue was purified by flash silica gel chromatography (ISCO®; 4 g SepaFlash® silica flash column, eluting at 30 mL / min with 0 - 5% MeOH / DCM) to give ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolan-2-yl)methyl butyl carbonate (7.6 mg, yield 11.3%) as a white solid. LCMS (ESI) m / z, C 17 H 20 FN5O5: calculated 393.14, found (M + H) + : 394.1. 11H NMR (400 MHz, CD3CN) δ (ppm): 7.96 (s, 1H), 6.27 (m, 3H), 4.73 (q, J = 6.8 Hz, 1H), 4.46 (d, J = 11.6 Hz, 1H), 4.31 - 4.23 (m, 1H), 4.10 - 3.99 (m, 2H), 3.75 - 3.66 (m, 1H), 2.99 (s, 1H), 2.85 (m, 1H), 2.57 (m, 1H), 1.61 - 1.52 (m, 2H), 1.28 (m, 2H), 0.94 - 0.87 (m, 3H); 19 19F NMR (376 MHz, CD3CN) δ (ppm): -52.86 (s, 1F).

[0265] Example 12: (2R,3S,5R)-5-(6-Amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-yl ((5-methyl-2-oxo-1,3-dioxol-4-yl)methyl) carbonate

Chem.

[0266] (2R,3S,5R)-5-(6-Amino-2-fluoro-9H-purin-9-yl)-2-(((tert-butyldimethylsilyl)oxy)methyl)-2-ethynyltetrahydrofuran-3-ol Preparation

Chem.

[0267] To a solution of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (0.5 g, 1.71 mmol, 1 equiv) in DMF (5 mL) were added imidazole (349 mg, 5.13 mmol, 3 equiv) and TBSCl (386 mg, 2.56 mmol, 1.5 equiv). The mixture was stirred at 25 °C for 16 h. The reaction mixture was concentrated. The resulting residue was purified by flash silica gel chromatography (ISCO (registered trademark); 4 g SepaFlash (registered trademark) silica flash column, eluting with 0 - 5% MeOH / DCM at 18 mL / min) to give (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-(((tert-butyldimethylsilyl)oxy)methyl)-2-ethynyltetrahydrofuran-3-ol (331 mg, 47.5% yield) as a pale yellow solid. 1 H NMR (400 MHz, CD3CN) δ (ppm) 8.03 (s, 1H), 6.39 - 6.17 (m, 3H), 4.71 (q, J = 6.8 Hz, 1H), 3.93 - 3.87 (m, 1H), 3.84 - 3.76 (m, 1H), 3.49 (d, J = 6.4 Hz, 1H), 2.91 (s, 1H), 2.85 - 2.73 (m, 1H), 2.55 (m, 1H), 2.50 (s, 2H), 0.88 - 0.82 (m, 9H), 0.02 (d, J = 16 Hz, 6H); 19 F NMR (376 MHz, CD3CN) δ (ppm) -53.04 (s, 1F).

[0268] Preparation of [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-2-[tert-butyl(dimethyl)silyl]oxymethyl]-2-ethynyl-tetrahydrofuran-3-yl] (4-nitrophenyl) carbonate

Chemical Structure

[0269] A solution (2 mL) of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-(((tert-butyldimethylsilyl)oxy)methyl)-2-ethynyltetrahydrofuran-3-ol (100 mg, 0.245 mmol, 1 equiv) in THF (2 mL) was added pyridine (0.40 mL, 20 equiv) and (4-nitrophenyl) carbonochloridate (494 mg, 2.45 mmol, 10 equiv). The mixture was stirred at 25 °C for 50 h. The reaction mixture was concentrated. The resulting residue was purified by flash silica gel chromatography (ISCO (registered trademark); 4 g SepaFlash (registered trademark) silica flash column, eluting with 0 - 5% MeOH / DCM at 18 mL / min) to give compound [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-2-[tert-butyl(dimethyl)silyl]oxymethyl]-2-ethynyl-tetrahydrofuran-3-yl] (4-nitrophenyl) carbonate (178 mg, crude) as a pale yellow solid.

[0270] Preparation of [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-2-[tert-butyl(dimethyl)silyl]oxymethyl]-2-ethynyl-tetrahydrofuran-3-yl] (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl carbonate

Chemical formula

[0271] A solution of [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-2-[tert-butyl(dimethyl)silyl]oxymethyl]-2-ethynyl-tetrahydrofuran-3-yl](4-nitrophenyl) carbonate (249 mg, 0.435 mmol, 1 equiv) in THF (4 mL) was added with DMAP (5 mg, 0.043 mmol, 0.1 equiv) and 4-(hydroxymethyl)-5-methyl-1,3-dioxol-2-one (170 mg, 1.30 mmol, 3 equiv). The mixture was stirred at 30 °C for 2 h. The reaction mixture was concentrated. The obtained residue was purified by flash silica gel chromatography (ISCO (registered trademark); 4 g SepaFlash (registered trademark) silica flash column, eluting at 18 mL / min with 0 - 5% MeOH / DCM) to give the compound [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-2-[tert-butyl(dimethyl)silyl]oxymethyl]-2-ethynyl-tetrahydrofuran-3-yl](5-methyl-2-oxo-1,3-dioxol-4-yl)methyl carbonate (239 mg, yield 97.6%) as a brown solid. 1 H NMR (400 MHz, CD3CN) δ (ppm): 8.06 - 7.95 (m, 1H), 6.42 - 6.26 (m, 3H), 5.61 (m, 1H), 4.97 (m, 2H), 3.98 - 3.93 (m, 1H), 3.89 - 3.83 (m, 1H), 3.16 - 3.03 (m, 1H), 2.95 (s, 1H), 2.70 (m, 1H), 2.15 (s, 3H), 0.86 (s, 9H), 0.04 (d, J = 10.3 Hz, 6H); 19 F NMR (376 MHz, CD3CN) δ (ppm): -52.66(s, 1F).

[0272] Preparation of (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-yl((5-methyl-2-oxo-1,3-dioxol-4-yl)methyl) carbonate

Chem.

[0273] A solution of [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-2-[tert-butyl(dimethyl)silyl]oxymethyl]-2-ethynyl-tetrahydrofuran-3-yl](5-methyl-2-oxo-1,3-dioxol-4-yl)methyl carbonate (359 mg, 0.637 mmol, 1 equiv) in pyridine (3.5 mL) and TEA (3.5 mL) was added with N,N-diethylethanamine; trifluoride (3.55 g, 22.02 mmol, 34.58 equiv) at 0 °C, and then the mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated. The obtained residue was purified by flash silica gel chromatography (ISCO®; 4 g of SepaFlash® silica flash column, eluted with 0-5% MeOH / DCM at 18 mL / min) to give (2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-yl ((5-methyl-2-oxo-1,3-dioxol-4-yl)methyl) carbonate (230 mg, yield 80%) as a white solid. LCMS (ESI) m / z, C 18 H 16 FN5O8: 449.1, found (M+H) + : 450.1. 1 H NMR (400 MHz, CD3CN) δ (ppm): 7.95 (s, 1H), 6.47 (s, 2H), 6.34 (m, 1H), 5.53 (m, 1H), 5.02 - 4.93 (m, 3H), 3.92 - 3.70 (m, 2H), 3.07 (m, 1H), 2.95 (s, 1H), 2.61 (m, 1H), 2.15 (s, 3H); 19 F NMR (376 MHz, CD3CN) δ (ppm): -53.27 (s, 1F).

[0274] Example 13: ((2R,3S,5R)-5-(6-Amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-((((5-methyl-2-oxo-1,3-dioxol-4-yl)methoxy)carbonyl)oxy)tetrahydrofuran-2-yl)methyl ((5-methyl-2-oxo-1,3-dioxol-4-yl)methyl) carbonate

Chem.

[0275] Preparation of [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-2-ethynyl-2-[(4-nitrophenoxy)carbonyloxymethyl]tetrahydrofuran-3-yl](4-nitrophenyl) carbonate

Chem.

[0276] To a solution of (2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-2-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3-ol (100 mg, 0.341 mmol, 1 equiv) in DCM (5 mL) were added pyridine (674 mg, 8.53 mmol, 25 equiv) and (4-nitrophenyl) carbonochloridate (1.03 g, 5.12 mmol, 15 equiv) at 26 °C. The mixture was stirred at 26 °C for 4 h. The mixture was quenched with H2O (30 mL). DCM (30 mL) was added to the mixture, and it was washed with H2O (30 mL) and brine (30 mL). The organic layer was dried over Na2SO4 and concentrated. The residue was purified by flash silica gel chromatography (ISCO®; 4 g SepaFlash® silica flash column, eluting with a 0 - 6% methanol / dichloromethane gradient at 18 mL / min) to give [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-2-ethynyl-2-[(4-nitrophenoxy)carbonyloxymethyl]tetrahydrofuran-3-yl] (4-nitrophenyl) carbonate (137 mg, yield 51%, purity 80%) as a white solid. LCMS (ESI) m / z, C 26 H 18 FN7O 11 : calculated 623.10, found 624.1 (M + H) + 。

[0277] Preparation of [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-2-ethynyl-2-[(5-methyl-2-oxo-1,3-dioxol-4-yl)methoxycarbonyloxymethyl]tetrahydrofuran-3-yl] (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl carbonate

Chemical formula

[0278] A solution of [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-2-ethynyl-2-[(4-nitrophenoxy)carbonyloxymethyl]tetrahydrofuran-3-yl] (4-nitrophenyl) carbonate (99 mg, 0.159 mmol, 1 equiv) in THF (5 mL) was added with DMAP (1.9 mg, 0.016 mmol, 0.1 equiv) and 4-(hydroxymethyl)-5-methyl-1,3-dioxol-2-one (248 mg, 1.91 mmol, 12 equiv). The mixture was stirred at 25 °C for 16 h. The mixture was quenched with H2O (30 mL). DCM (30 mL) was added to the mixture. The mixture was washed with H2O (30 mL) and brine (30 mL). The organic layer was dried over Na2SO4 and concentrated. The resulting residue was purified by flash silica gel chromatography (ISCO®; 4 g SepaFlash® silica flash column, eluting with a 0 - 10% methanol / dichloromethane gradient at 18 mL / min) to give [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-2-ethynyl-2-[(5-methyl-2-oxo-1,3-dioxol-4-yl)methoxycarbonyl]tetrahydrofuran-3-yl] (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl carbonate (89 mg, yield 56%, purity 60%). The substance was further purified by flash silica gel chromatography and recrystallized (CH3CN / Hep) to give [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-2-ethynyl-2-[(5-methyl-2-oxo-1,3-dioxol-4-yl)methoxycarbonyloxymethyl]tetrahydrofuran-3-yl] (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl carbonate as a white solid. LCMS (ESI) m / z, C 24 H 20 FN5O 13 : Calcd 605.10, found (M + H) + : 606.1. 11H NMR (400 MHz, DMSO-d6) δ (ppm) 8.30 (s, 1H), 7.89 (br s, 2H), 6.38 - 6.34 (m, 1H), 5.62 - 5.59 (m, 1H), 5.14 - 5.06 (m, 2H), 4.99 (s, 2H), 4.53 (d, J = 11.6 Hz, 1H), 4.37 (d, J = 11.6 Hz, 1H), 3.83 (s, 1H), 3.21 - 3.17 (m, 1H), 2.73 - 2.70 (m, 1H), 2.19 (s, 3H), 2.14 (s, 3H). 19 19F NMR (376 MHz, DMSO-d6) δ (ppm) -51.47 (s, 1F).

[0279] Example 14: Conversion and Stability of Adenosine Derivative Prodrug In both plasma and liver S9 assays, the stability of the prodrug and the conversion of the prodrug to the parent EFdA (Formula T-1A) were measured, and the data are shown in Table 2.

[0280] Plasma Stability Pooled frozen plasma was thawed in a 37 °C water bath before the experiment. The plasma was centrifuged at 4000 rpm for 5 minutes to remove any clots. Adjust the pH to 7.4 ± 0.1 if necessary. Preparation of test compound and positive control (propantheline bromide): A 1 mM intermediate solution was prepared by diluting 10 μL of the stock solution with 90 μL of MeOH, and a 1 mM intermediate solution of the positive control propantheline was prepared by diluting 10 μL of the stock solution with 90 μL of ultrapure water. A 100 μM dosing solution was prepared by diluting 20 μL of the intermediate solution (1 mM) with 180 μL of MeOH. 98 μL of blank plasma was spiked with 2 μL of the dosing solution (100 μM) to achieve a final concentration of 2 μM in duplicate, and the samples were incubated in a water bath at 37 °C. At each time point (0, 10, 30, 60, and 120 minutes), 400 μL of stop solution (0.1% FA in MeOH containing 200 ng / mL tolbutamide and 200 ng / mL labetalol) was added to precipitate the protein and mixed well. The sample plate was centrifuged at 4,000 rpm for 10 minutes. An aliquot (100 μL) of the supernatant was transferred from each well to another plate. Data analysis: The percentage of the remaining test compound after incubation in plasma was calculated using the following formula: Percentage remaining = 100 x (PAR at specified incubation time / PAR at T0 time) where PAR is the peak area ratio of the analyte to the internal standard (IS) (LC / MS / MS mobile phase conditions: 0.1% formic acid in water / 0.1% formic acid in acetonitrile. The specified incubation time points are T0 (0 minutes), Tn (n = 0, 10, 30, 60, 120 minutes).

[0281] Liver S9 stability Intermediate solution: Dilute 5 μL of the compound or control (7-ethoxycoumarin) from the stock solution (10 mM) with 495 μL of MeOH (concentration: 100 μM, 1% DMSO, 99% MeOH). Stop solution: Cold ACN (containing tolbutamide and labetalol at 100 ng / mL as internal standards). Add 2 μL of the test compound or control working solution / well to all plates except the matrix blank (T0, T5, T10, T20, T30, T60, NCF60). Add 600 μL / well of the stop solution (cooled to 4°C, containing 100 ng / mL of tolbutamide / 100 ng / mL of labetalol), finish the T0 plate, and place it on ice. Dispense 840 μL / well of the S9 solution into a 96-well plate as a reservoir according to the plate map. Next, add 100 μL / well to all plates by Apricot. Incubate the S9 solution and the compound at 37°C for approximately 10 minutes, except for NCF60 and T0. After adding the S9 solution and 98 μL of PB buffer to NCF60, incubate at 37°C without preheating and start Timer 1. After 60 minutes, add 600 μL / well of the stop solution to terminate the reaction. After preheating, dispense 760 μL / well of the cofactor solution into a 96-well plate as a reservoir according to the plate map. Next, add 98 μL / well to each plate by Apricot to initiate the reaction. Incubate at 37°C, start Timer 2, add 600 μL / well of the stop solution (4°C, containing 100 ng / mL of tolbutamide and labetalol) to terminate the reaction. Centrifuge the samples at 4000 rpm for 20 minutes. While centrifuging, fill eight new 96-well plates with 300 μL of HPLC water, then transfer 100 μL of the supernatant and mix it with the water for LC / MS / MS, and transfer it to Bioanalytical Services for LC-MS / MS analysis. Use the equation of the first-order rate law to calculate t 1 / 2 and CL: Equation of the first-order rate law: [Number] CL int(S9) =Vd·k e Vd = 1 mL / mg The stability results of exemplary compounds in human plasma and human liver S9 are listed in Table 2 below. [Table 2]

[0282] The data showed that in the human plasma and liver S9 assays, the adenosine derivatives 4-A and 4-C could be efficiently converted to the target drug, and in the liver S9 assay, 6-A could be efficiently converted to the target drug.

[0283] Example 15: Plasma Exposure after Oral Administration of Prodrug to Beagle Dogs After oral administration of an EFdA dose of 5 mg equivalent / kg, the pharmacokinetics of EFdA and the prodrug formula 4-A were tested in dogs.

[0284] Formulation: The prodrug was formulated as a 1.65 mg / mL solution in 20% aqueous PEG400 within 0.5 hours before administration.

[0285] Dose Administration and Sample Collection: The in-life phase of this study was conducted at Charles River Laboratory (CRL) in Worcester, MA, in accordance with the Institutional Animal Care and Use Committee (IACUC) guidelines that comply with the Animal Welfare Act and the Guide for the Care and Use of Laboratory Animals, along with the standard animal treatment methods of the CRL Animal Experimentation Committee, and was approved by the IACUA committee. Fasted male beagle dogs (10 + / - 2 kg) were used in the study. Each drug was administered as a single dose by co-oral administration (5 ml / kg). The prodrug formula 4-A dose (8.25 mg / kg) was equivalent to a dose of 5 mg / kg of EFdA. Plasma samples were collected at 0 (pre-dose), 30 minutes, 1, 2, 4, 6, 8, 12, and 24 hours after administration. Blood (approximately 0.1 - 0.2 mL) was immediately processed into plasma by centrifuging at 3,500 rpm for 10 minutes at 5°C immediately after collection. Plasma samples were frozen and maintained at -70°C until analysis. To stabilize the prodrug during sample collection and subsequent analysis, the following stabilizing reagents were added to the K2EDTA tubes for blood collection on wet ice before sample collection: For every 100 mL of blood, the 15 mL pre-made inhibitor cocktail consisted of 1 mM DFP, 100 mM dichlorvos, 100 mM 2-hydroxyquinoline, 100 mM PCMB, 1 mM paraoxon, 100 mM PMSF, 100 mM NaF, 30.0 mM EDTA, and 15 mM citric acid, 10 mL of 0.2 M eserine, and 10 mL of 0.2 M BNPP solution.

[0286] Determination of EFdA and Prodrug in Plasma: Briefly, plasma (20 μL) was mixed with 100 μL of acetonitrile to precipitate proteins. The same cocktail protocol was added as in the sample collection procedure to stabilize the prodrug in the standard and QC samples.

[0287] Bioanalysis: For the quantitative analysis of plasma samples, a Sciex API-6500 triple quadrupole mass spectrometer coupled with a Shimadzu HPLC system (Framingham, MA 01701) was used. The column was a Waters HSS T3 column (2.1×50 mm, 1.8 mm). The mobile phases used were as follows: A, 5% acetonitrile in 2 mM ammonium formate buffer; B, 95% acetonitrile in 2 mM ammonium formate buffer, pH 6.0. The flow rate was 0.6 mL / min and the total run time was 3.0 minutes. The HPLC gradient was increased linearly from 98% A / 2% B for 0.20 minutes, then to 25% over the next 1.40 minutes, then increased to 100% mobile phase B over the next 1.0 minute, then held for an additional 0.2 minutes, and then the mobile phase B was decreased to 2% within the next 0.2 minutes. Detection of the prodrug and EFdA was achieved using positive ion electrospray mass spectrometry mode in unit resolution mode. To quantify both the prodrug and EFdA, multiple reaction monitoring (MRM) mode was used. For example, the MRM transition for EFdA was 294.0~153.90 Da and the transition for the prodrug 4-A was 450.0~153.9 Da. Peak areas were integrated by the Sciex program Analyst® version 1.6.3 operating on a Windows® 7 computer, where the concentration was determined by weighted (1 / x2) linear regression of the peak area ratio (peak area of EFdA / corresponding IS peak area) against the nominal concentration of the plasma calibration standard. Calculations were performed on unrounded numerical values. Overall, Analyst® determined the accuracy and precision for the calibration standards and QC samples.

[0288] Pharmacokinetic calculations: Non-compartmental (NCA) analysis of the individual plasma concentration-time data of EFdA and the prodrug was performed using the WinNonlin module (version 8.3.0.5005, Certara Inc., Princeton, NJ 08540) in the PK / PD platform. Calculations were performed before rounding values, and nominal sampling times were used for pharmacokinetic analysis. Exposure was expressed as the area under the concentration curve (AUC 0~24時間 ) in plasma from zero to 24 hours. AUC values were calculated using the linear trapezoidal rule.

[0289] Plasma concentration: The results of the PK studies are shown in Tables 3 and 4. These data establish in vivo that prodrug formula 4-A can efficiently release EFdA in vivo, with the prodrug being readily delivered orally and minimal prodrug detected in the systemic circulation. For example, prodrug formula 4-A can release significantly more EFdA than an equivalent dose of EFdA, namely 91%, 102%, 55%, 79%, and 200% more at the time points of 0.25, 0.5, 1, 2, and 4 hours, respectively (see Table 3). Further, prodrug formula 4-A can result in a higher AUC and C max than an equivalent dose of EFdA (see Table 4). [Table 3-1] [Table 3-2] [Table 4]

[0290] Numbered embodiments of the present disclosure Other subjects contemplated by the present disclosure are described in the following numbered embodiments. 1. An adenosine derivative having the formula (1), or a pharmaceutically acceptable salt, stereoisomer, tautomer, or solvate thereof, wherein [Chemical formula] In the formula, R 1 , R 1’ , and R 2 are each independently -H, -C(O)N(R 3 )(R 3’ ), -C(O)OR 4 , -R 5 , -L 1 -R 5 , or Z-L 4 -R 5 , and at least one of R 1 and R 2 is not -H, R 3 , R 3’ , and R 4 are each independently -H, C1-C10 alkyl, C2-C10 alkenyl, C3-C10 cycloalkyl, 3-10 membered heterocycloalkyl, aryl, or heteroaryl, R 5 is

Chemical formula

[0291] 2. The adenosine derivative is represented by formula (2):

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0292] 3. The adenosine derivative is represented by formula (2-A):

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chem.

Chem.

Chem.

Chem.

[0293] 4. The adenosine derivative according to Embodiment 1, wherein the adenosine derivative is ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolane-2-yl)methyl isopropylcarbamate.

[0294] 5. The adenosine derivative according to Embodiment 1, wherein the adenosine derivative is isopropyl (9-((2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)oxolane-2-yl)-2-fluoro-9H-purin-6-yl)carbamate.

[0295] 6. The adenosine derivative according to Embodiment 1, wherein the adenosine derivative is ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolane-2-yl)methyl ((5-methyl-2-oxo-1,3-dioxol-4-yl)methyl) carbonate.

[0296] 7. The adenosine derivative according to Embodiment 1, wherein the adenosine derivative is ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolane-2-yl)methyl methylcarbamate.

[0297] 8. The adenosine derivative according to Embodiment 1, wherein the adenosine derivative is 4-(((9-((2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)oxolane-2-yl)-2-fluoro-9H-purin-6-yl)amino)methyl)-5-methyl-1,3-dioxolan-2-one.

[0298] 9. The adenosine derivative according to Embodiment 1, wherein the adenosine derivative is ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolane-2-yl)methyl (2-(methyl((5-methyl-2-oxo-1,3-dioxolan-4-yl)methyl)amino)ethyl)carbamate.

[0299] 10. The adenosine derivative according to Embodiment 1, wherein the adenosine derivative is [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-2-ethynyl-3-hydroxy-oxolane-2-yl]methyl 4-[methyl-[(5-methyl-2-oxo-1,3-dioxolan-4-yl)methoxycarbonyl]amino]butanoate.

[0300] 11. The R 5 , -L 1 -R 5 , or -Z-L 4 -R 5 is selected from Formulas 9 to 24, the adenosine derivative according to Embodiment 1.

Chemical Formula

[0301] 12. The adenosine derivative according to any one of Embodiments 1 to 11, wherein the adenosine derivative includes reverse transcriptase inhibitor activity in vivo, reverse transcriptase chain terminator activity in vivo, DNA translocation inhibitor activity in vivo, or a combination thereof.

[0302] 13. A pharmaceutical composition comprising an adenosine derivative having the formula (1),

Chemical formula

Chemical formula

[0303] 14. The adenosine derivative is of formula (2):

Chem.

Chem.

Chem.

Chem.

Chem.

Chem.

Chem.

Chem.

[0304] 15. The adenosine derivative is of formula (2-A):

Chem.

Chem.

Chem.

Chem.

Chem.

Chem.

Chem.

Chem.

[0305] 16. The pharmaceutical composition according to embodiment 13, wherein the adenosine derivative comprises ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxy tetrahydrofuran-2-yl)methyl isopropylcarbamate, or a pharmaceutically acceptable salt thereof.

[0306] 17. The pharmaceutical composition according to embodiment 13, wherein the adenosine derivative comprises isopropyl (9-((2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2-fluoro-9H-purin-6-yl)carbamate, or a pharmaceutically acceptable salt thereof.

[0307] 18. The pharmaceutical composition according to embodiment 13, wherein the adenosine derivative is ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolane-2-yl)methyl ((5-methyl-2-oxo-1,3-dioxol-4-yl)methyl) carbonate, or a pharmaceutically acceptable salt thereof.

[0308] 19. The pharmaceutical composition according to embodiment 13, wherein the adenosine derivative is ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolane-2-yl)methyl methylcarbamate, or a pharmaceutically acceptable salt thereof.

[0309] 20. The pharmaceutical composition according to embodiment 13, wherein the adenosine derivative is 4-(((9-((2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)oxolane-2-yl)-2-fluoro-9H-purin-6-yl)amino)methyl)-5-methyl-1,3-dioxolan-2-one, or a pharmaceutically acceptable salt thereof.

[0310] 21. The pharmaceutical composition according to embodiment 13, wherein the adenosine derivative is ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolane-2-yl)methyl (2-(methyl((5-methyl-2-oxo-1,3-dioxol-4-yl)methyl)amino)ethyl) carbamate, or a pharmaceutically acceptable salt thereof.

[0311] 22. The pharmaceutical composition according to embodiment 13, wherein the adenosine derivative is [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-2-ethynyl-3-hydroxy-oxolane-2-yl]methyl 4-[methyl-[(5-methyl-2-oxo-1,3-dioxol-4-yl)methoxycarbonyl]amino]butanoate, or a pharmaceutically acceptable salt thereof.

[0312] 23. The said R 5 , -L 1 , -R 5 , or -Z-L 4 , -R 5 is selected from Formulas 9 to 24, the pharmaceutical composition according to Embodiment 13. [Chemical formula]

[0313] 24. The pharmaceutical composition according to any one of Embodiments 13 to 23, further comprising a pharmaceutically acceptable carrier.

[0314] 25. An effective dosage of abacavir, abacavir sulfate, lamivudine, amprenavir, atazanavir sulfate, AZT, vicriviroc, cabotegravir, darunavir, didanosine, dideoxyinosine, dolutegravir, delavirdine, efavirenz, emtricitabine, tenofovir disoproxil fumarate, tenofovir alafenamide, 4'-ethynyl-2-fluoro-2'-deoxyadenosine, elvitegravir, etravirine, fosamprenavir calcium, indinavir, indinavir sulfate, lamivudine, lopinavir, a combination of lopinavir and ritonavir, darunavir, a combination of darunavir and cobicistat, maraviroc, nelfinavir, nelfinavir mesylate, nevirapine, PPL-100, raltegravir, rilpivirine, stavudine, tipranavir, vicriviroc, or a combination thereof, the pharmaceutical composition according to any one of Embodiments 13 to 24, further comprising one or more HIV antiviral agents selected therefrom.

[0315] 26. A method for treating a disease, comprising administering to a subject in need of treatment an effective dosage of a pharmaceutical composition comprising an adenosine derivative having Formula (1), or a pharmaceutically acceptable salt, stereoisomer, tautomer, or solvate thereof, [Chemical formula] wherein, R 1 、R 1’ 、and R 2 are each, independently, -H, -C(O)N(R 3 )(R 3’ ), -C(O)OR 4 , -R 5 , -L 1 -R 5 , or Z-L 4 -R 5 , where R 1 and R 2 at least one of which is not -H, R 3 , R 3’ and R 4 are each, independently, -H, C1-C10 alkyl, C2-C10 alkenyl, C3-C10 cycloalkyl, 3-10 membered heterocycloalkyl, aryl, or heteroaryl, R 5 is,

Chemical formula

[0316] 27. The adenosine derivative is of formula (2):

Chemical formula

[0317] 28. The adenosine derivative is Formula (2-A): [Chemical formula] Formula (3-A): [Chemical formula] Formula (4-A): [Chemical formula] Formula (5-A): [Chemical formula] Formula (6-A): [Chemical formula] Formula (7-A): [Chem.] Formula (8-A): [Chem.] Formula (4-C): [Chem.] The adenosine derivative according to embodiment 26, which has or is a pharmaceutically acceptable salt, stereoisomer, tautomer, or solvate thereof.

[0318] 29. The method according to embodiment 26, wherein the adenosine derivative comprises ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolane-2-yl)methyl isopropylcarbamate, or a pharmaceutically acceptable salt thereof.

[0319] 30. The method according to embodiment 26, wherein the adenosine derivative comprises isopropyl (9-((2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)oxolane-2-yl)-2-fluoro-9H-purin-6-yl)carbamate, or a pharmaceutically acceptable salt thereof.

[0320] 31. The method according to embodiment 26, wherein the adenosine derivative comprises ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolane-2-yl)methyl ((5-methyl-2-oxo-1,3-dioxol-4-yl)methyl) carbonate, or a pharmaceutically acceptable salt thereof.

[0321] 32. The method according to embodiment 26, wherein the adenosine derivative comprises ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolane-2-yl)methyl methylcarbamate, or a pharmaceutically acceptable salt thereof.

[0322] 33. The method according to embodiment 26, wherein the adenosine derivative comprises 4-(((9-((2R,4S,5R)-5-ethynyl-4-hydroxy-5-(hydroxymethyl)oxolane-2-yl)-2-fluoro-9H-purin-6-yl)amino)methyl)-5-methyl-1,3-dioxolan-2-one, or a pharmaceutically acceptable salt thereof.

[0323] 34. The method according to embodiment 26, wherein the adenosine derivative is ((2R,3S,5R)-5-(6-amino-2-fluoro-9H-purin-9-yl)-2-ethynyl-3-hydroxyoxolane-2-yl)methyl (2-(methyl((5-methyl-2-oxo-1,3-dioxolan-4-yl)methyl)amino)ethyl)carbamate, or a pharmaceutically acceptable salt thereof.

[0324] 35. The method according to embodiment 26, wherein the adenosine derivative is [(2R,3S,5R)-5-(6-amino-2-fluoro-purin-9-yl)-2-ethynyl-3-hydroxy-oxolane-2-yl]methyl 4-[methyl-[(5-methyl-2-oxo-1,3-dioxolan-4-yl)methoxycarbonyl]amino]butanoate, or a pharmaceutically acceptable salt thereof.

[0325] 36. The adenosine derivative is the R 5 , -L 1 , -R 5 , or -Z-L selected from formulas 9 to 24 4 , -R 5 The method according to embodiment 26 comprising.

Chemical formula

[0326] 37. The method according to any one of embodiments 26 - 36, wherein the pharmaceutical composition is administered to the subject via intramuscular (IM) injection, subcutaneous (SC) injection, intravenous (IV) injection, oral administration, topical application, implant application, or a combination thereof.

[0327] 38. The method further comprises measuring a sample of the subject to determine a measured level of a target drug in the sample, wherein the target drug has the formula (T - 1),

Chem.

[0328] 39. The target drug has the formula (T - 1A),

Chem.

[0329] 40. The target drug is (2R,3S,5R)-5-(6 - amino - 2 - fluoro - 9H - purin - 9 - yl)-2 - ethynyl - 2-(hydroxymethyl)tetrahydrofuran - 3 - ol, or a pharmaceutically acceptable salt thereof, according to embodiment 38.

[0330] 41. When the measured level of the target drug is different from a predetermined target level of the target drug, further comprising adjusting the effective dosage to produce a modified effective dosage, and administering the modified effective dosage to the subject, according to any one of embodiments 38 - 40.

[0331] 42. The method according to any one of embodiments 26 to 41, wherein the disease is acquired immunodeficiency syndrome (AIDS), wild-type HIV-1, NRTI-resistant HIV-1, HIV-2, HIV having the M184V mutation, HIV having K65R, or multi-drug resistant HIV.

[0332] 43. The method according to any one of embodiments 26 to 42, further comprising administering to the subject an effective dosage of one or more anti-HIV agents selected from abacavir, abacavir sulfate, lamivudine, amprenavir, atazanavir sulfate, AZT, vicriviroc, cabotegravir, darunavir, didanosine, dideoxyinosine, dolutegravir, delavirdine, efavirenz, emtricitabine, tenofovir disoproxil fumarate, tenofovir alafenamide, 4'-ethynyl-2-fluoro-2'-deoxyadenosine, elvitegravir, etravirine, fosamprenavir calcium, indinavir, indinavir sulfate, lamivudine, lopinavir, the combination of lopinavir and ritonavir, darunavir, the combination of darunavir and cobicistat, maraviroc, nelfinavir, nelfinavir mesylate, nevirapine, PPL-100, raltegravir, rilpivirine, stavudine, tipranavir, vicriviroc, or combinations thereof.

[0333] 44. The method according to embodiment 43, wherein the adenosine derivative and the one or more anti-HIV agents are administered to the subject together or separately via oral administration, parenteral administration, or a combination thereof.

[0334] 45. The method according to embodiment 44, wherein the adenosine derivative and the one or more anti-HIV agents are administered to the subject on a daily, weekly, bi-weekly, or monthly dosing schedule.

[0335] 46. Use of a pharmaceutical composition according to any one of Embodiments 13 to 25 for treating a disease in a subject in need of treatment of the disease, wherein the disease is acquired immunodeficiency syndrome (AIDS), wild-type HIV-1, NRTI-resistant HIV-1, HIV-2, HIV having the M184V mutation, HIV having K65R, or multi-drug resistant HIV.

[0336] 47. Use of a method according to any one of Embodiments 26 to 45 for treating a disease in a subject in need of treatment of the disease, wherein the disease is acquired immunodeficiency syndrome (AIDS), wild-type HIV-1, NRTI-resistant HIV-1, HIV-2, HIV having the M184V mutation, HIV having K65R, or multi-drug resistant HIV.

[0337] 48. Use of an adenosine derivative according to any one of Embodiments 1 to 12 for the manufacture of a medicament for treating a disease, wherein the disease is acquired immunodeficiency syndrome (AIDS), wild-type HIV-1, NRTI-resistant HIV-1, HIV-2, HIV having the M184V mutation, HIV having K65R, or multi-drug resistant HIV.

[0338] 49. A method for preventing infection in a subject in need of prevention of infection, the method comprising administering to the subject an effective dosage of a pharmaceutical composition according to any one of Embodiments 13 to 25, wherein the subject does not have detectable symptoms of the infection.

[0339] 50. The method according to Embodiment 49, wherein the infectious disease comprises a disease selected from acquired immunodeficiency syndrome (AIDS), wild-type HIV-1, NRTI-resistant HIV-1, HIV-2, HIV having the M184V mutation, HIV having K65R, multi-drug resistant HIV, infectious diseases, or combinations thereof.

[0340] 51. The method according to embodiment 50, wherein the detectable symptoms include symptoms of acquired immunodeficiency syndrome (AIDS), wild-type HIV-1, NRTI-resistant HIV-1, HIV-2, HIV having the M184V mutation, HIV having K65R, symptoms of infection with an HIV virus including multi-drug resistant HIV, or combinations thereof.

[0341] 52. The method according to embodiment 51, wherein the pharmaceutical composition is administered to the subject on a daily, weekly, bi-weekly, or monthly dosing schedule.

[0342] 53. An effective dosage of abacavir, abacavir sulfate, lamivudine, amprenavir, atazanavir sulfate, AZT, vicriviroc, cabotegravir, darunavir, didoxythymidine, didoxinosine, dolutegravir, delavirdine, efavirenz, emtricitabine, tenofovir disoproxil fumarate, tenofovir alafenamide, 4'-ethynyl-2-fluoro-2'-deoxyadenosine, elvitegravir, etravirine, fosamprenavir calcium, indinavir, indinavir sulfate, lamivudine, lopinavir, the combination of lopinavir and ritonavir, darunavir, the combination of darunavir and cobicistat, maraviroc, nelfinavir, nelfinavir mesylate, nevirapine, PPL-100, raltegravir, rilpivirine, stavudine, tipranavir, vicriviroc, or combinations thereof, further comprising administering to the subject one or more anti-HIV agents selected therefrom. The method according to embodiment 52.

[0343] 54. The method according to embodiment 53, wherein the one or more anti-HIV agents are administered to the subject together with or separately from the pharmaceutical composition.

[0344] 55. A method for treating HIV infection, comprising administering to a subject in need of treatment an effective dosage of the pharmaceutical composition according to any one of embodiments 1 to 25. The method.

[0345] 56. A method for preventing HIV infection, comprising administering to a subject in need of prevention a pharmaceutical composition according to any one of embodiments 1 to 25 in an effective dosage amount, said method.

[0346] 57. The method according to embodiment 55 or 56, wherein the HIV infection is caused by wild-type HIV-1, NRTI-resistant HIV-1, HIV-2, HIV having the M184V mutation, HIV having K65R, or multi-drug resistant HIV.

[0347] 58. The method according to any one of embodiments 55 to 57, wherein the administration is by oral administration.

[0348] 59. The method according to any one of embodiments 55 to 57, wherein the administration is by parenteral administration.

[0349] 60. The method according to embodiment 59, wherein the parenteral administration is by intramuscular or subcutaneous injection.

[0350] 61. The method according to any one of embodiments 55 to 60, wherein the administration of the pharmaceutical composition results in a higher plasma concentration of EFdA compared to the administration of an equivalent dose of EFdA under the same conditions.

[0351] 62. The method according to embodiment 61, wherein the administration of the pharmaceutical composition results in a plasma concentration of EFdA that is at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or 200% higher when compared to the administration of an equivalent dose of EFdA under the same conditions.

[0352] 63. The method according to embodiment 61, wherein the administration of the pharmaceutical composition results in a plasma concentration of EFdA that is 50% - 80%, 50% - 100%, or 50% - 200% higher when compared to the administration of an equivalent dose of EFdA under the same conditions.

[0353] 64. The method according to any one of embodiments 55 to 63, wherein the administration of the pharmaceutical composition results in a sustained release of EFdA when compared to the administration of an equivalent dose of EFdA under the same conditions.

[0354] 65. The method according to any one of embodiments 55 to 64, wherein the administration of the pharmaceutical composition results in a higher AUC of EFdA when compared to the administration of an equivalent dose of EFdA under the same conditions.

[0355] 66. The method according to embodiment 65, wherein the administration of the pharmaceutical composition results in an AUC of EFdA that is at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% higher when compared to the administration of an equivalent dose of EFdA under the same conditions.

[0356] 67. The method according to embodiment 65, wherein the administration of the pharmaceutical composition results in an AUC of EFdA that is 50% - 200%, 50% - 150%, or 80% - 120% higher when compared to the administration of an equivalent dose of EFdA under the same conditions.

[0357] 68. The method according to any one of embodiments 55 to 67, wherein the administration of the pharmaceutical composition results in a higher C max of EFdA when compared to the administration of an equivalent dose of EFdA under the same conditions.

[0358] 69. The method according to embodiment 68, wherein the administration of the pharmaceutical composition results in a C max of EFdA that is at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% higher when compared to the administration of an equivalent dose of EFdA under the same conditions.

[0359] 70. The method according to embodiment 68, wherein the administration of the pharmaceutical composition results in a C max of EFdA that is 50% - 200%, 50% - 150%, or 80% - 100% higher when compared to the administration of an equivalent dose of EFdA under the same conditions.

Claims

1. An adenosine derivative of formula (4-A), or a pharmaceutically acceptable salt thereof. 【Chemical 1】

2. An adenosine derivative of formula (6-A), or a pharmaceutically acceptable salt thereof. [Chemical 2]

3. An adenosine derivative of formula (4-C), or a pharmaceutically acceptable salt thereof. 【Chemical Formula 3】

4. An adenosine derivative according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, a pharmaceutical composition comprising.

5. The pharmaceutical composition according to claim 4, wherein the pharmaceutical composition is suitable for oral administration.

6. The pharmaceutical composition according to claim 4 or 5, wherein the pharmaceutical composition is in a liquid dosage form.

7. The pharmaceutical composition according to claim 6, wherein the liquid dosage form is a solution, an emulsion or a liquid suspension.

8. The pharmaceutical composition according to claim 4 or 5, wherein the pharmaceutical composition is in a solid dosage form.

9. The pharmaceutical composition according to claim 8, wherein the solid dosage form is a tablet, a capsule, a granule, a powder, a sachet, a reconstitutable powder, an inhalable dry powder, or a chewable tablet.

10. A pharmaceutical composition according to any one of claims 4 to 9 for use in the treatment of HIV infection.

11. A pharmaceutical composition according to any one of claims 4 to 9 for use in the prevention of HIV infection.

12. The pharmaceutical composition according to claim 10 or 11, wherein the HIV infection is caused by wild-type HIV-1, NRTI-resistant HIV-1, HIV-2, HIV having the M184V mutation, HIV having K65R, or multi-drug resistant HIV.

13. The pharmaceutical composition according to claim 10 or 11, wherein the pharmaceutical composition is administered by oral administration.

14. The pharmaceutical composition according to claim 10 or 11, wherein the administration of the pharmaceutical composition results in a higher plasma concentration of EFdA when compared to the administration of an equivalent dose of EFdA under the same conditions.

15. The pharmaceutical composition according to claim 10 or 11, wherein the administration of the pharmaceutical composition results in a sustained release of EFdA when compared to the administration of an equivalent dose of EFdA under the same conditions.

16. The pharmaceutical composition according to claim 10 or 11, wherein the administration of the pharmaceutical composition results in a higher AUC of EFdA when compared to the administration of an equivalent dose of EFdA under the same conditions.

17. When the administration of the pharmaceutical composition results in a higher C of EFdA when compared to the administration of an equivalent dose of EFdA under the same conditions ｍａｘ The pharmaceutical composition according to claim 10 or 11 Composition.

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