β-D-2'-deoxy-2'-α-fluoro-2'-β-C-substituted-2-modified-N6-substituted purine nucleotides for HCV treatment

Abstract

To provide compounds, pharmaceutical compositions, and methods and uses to treat and / or prevent infections of HCV.SOLUTION: The present invention provides a compound with a structure (A) containing a specific substituent, or a pharmaceutically acceptable salt thereof, or a composition thereof.SELECTED DRAWING: Figure 4

Description

[Technical Field]

[0001] Priority This application claims priority to U.S. Patent Application No. 62 / 129,319 filed 6 March 2015, U.S. Patent Application No. 62 / 253,958 filed 11 November 2015, and U.S. Patent Application No. 62 / 276,597 filed 8 January 2016, the entirety of which constitutes part of this specification by reference to each of them.

[0002] This invention relates to nucleotide compounds and compositions for treating hepatitis C virus ("HCV"), and to the use thereof. [Background technology]

[0003] Hepatitis C virus (HCV) is an RNA single-stranded virus and a member of the hepacivirus genus. It is estimated that 75% of all liver disease cases are caused by HCV. HCV infection can lead to cirrhosis and liver cancer, and if left untreated, can result in liver failure that may require a liver transplant. Approximately 170 to 200 million people worldwide are infected, with an estimated 3 to 4 million infected in the United States.

[0004] RNA polymerase is a key component in the targeting of single-stranded RNA viruses. The non-structural protein NS5B RNA-dependent RNA polymerase of HCV is the primary enzyme responsible for initiating and catalyzing viral RNA synthesis. Consequently, HCV NS5B is currently an attractive target for the discovery and development of anti-HCV drugs. NS5B inhibitors fall into two main subclasses: anabolized nucleoside analogs, which are assimilated into active triphophenes that act as alternative substrates for polymerase, and non-nucleoside inhibitors (NNIs), which bind to allosteric regions on the protein. Nucleoside or nucleotide inhibitors mimic natural polymerase substrates and act as chain arresters. They inhibit the initiation of RNA transcription and the initial elongation of the RNA chain.

[0005] In addition to targeting RNA polymerase, other RNA viral proteins may also be targeted in combination therapies. For example, HCV proteins that are further targets of therapeutic approaches are NS3 / 4A (serine proteases) and NS5A (non-structural proteins that are essential components of HCV replicases and exert a range of effects on cellular pathways).

[0006] In December 2013, sofosbuvir (Sovaldi®, Gilead Sciences), the first nucleoside NS5B polymerase inhibitor, was approved. It is a uridine phosphoramidate prodrug taken up by hepatocytes, and after intracellular activation, it delivers the active metabolite 2'-deoxy-2'-α-fluoro-β-C-methyluridine-5'-triphosphate. See the structure below. [ka]

[0007] Sovaldi® is the first drug to be proven safe and effective in treating certain types of HCV infection without requiring co-administration with interferon. Sovaldi® is the third drug to receive Breakthrough Therapy designation from the FDA.

[0008] In 2014, the U.S. FDA approved Harvoni® (ledipasvir, an NS5A inhibitor, and sofosbuvir) to treat chronic hepatitis C virus genotype 1 infection. Harvoni® was the first combination pill approved for the treatment of chronic HCV genotype 1 infection. It was also the first approved treatment regimen that did not require administration with interferon or ribavirin. Furthermore, the FDA approved simeprevir® (Olisio®) in combination with sofosbuvir® (Sovaldi®) as an all-oral, once-daily treatment for adults with genotype 1 HCV infection that does not contain interferon or ribavirin.

[0009] In 2014, the U.S. FDA also approved AbbVie's Viekira Pak®, a combination pack containing dasabuvir (a non-nucleoside NS5B polymerase inhibitor), ombitasvir (an NS5A inhibitor), paritaprevir (an NS3 / 4A inhibitor), and ritonavir. Viekira Pak® can be used with or without ribavirin to treat patients with genotype 1 HCV infection, including patients with compensated cirrhosis. Viekira Pak® does not require combination therapy with interferon.

[0010] In July 2015, the U.S. FDA approved Technivi® and Daklinza®, respectively, for the treatment of HCV genotype 4 and HCV genotype 3. Technivi® (ombitasvir / paritaprevir / ritonavir) is approved in combination with ribavirin for the treatment of HCV genotype 4 in patients without scarring and cirrhosis, and is a first-line treatment for HCV-4 infected patients that do not require co-administration of interferon. Daklinza® is approved for use in combination with Sovaldi® for the treatment of HCV genotype 3 infection. Daklinza® has demonstrated safety and efficacy in the treatment of HCV genotype 3 that does not require co-administration of interferon or ribavirin. It was the first drug to be developed.

[0011] In October 2015, the U.S. FDA warned that the HCV treatments Viekira Pak and Tecnivi may cause serious liver injury, primarily in patients with underlying advanced liver disease, and requested that additional safety information be added to the labels.

[0012] Other currently approved treatments for HCV include interferon alpha-2b or pegylated interferon alpha-2b (PegIntron®), sometimes administered with ribavirin (Rebetol®), NS3 / 4A telaprevir (Insivec®, Vertex, and Johnson & Johnson), and boceprevir (Victorelis®). , Merck), Simeprevir (Olisio (trademark), Johnson & Johnson), Paritaprevir (AbbVie), Ombitasvir (AbbVie), (NNI) Dasabuvir (ABT-333), and Merck's Zepatia (trademark) (a combination of two drugs, grazoprevir and elbasvir in single tablets) (Combinations) are mentioned.

[0013] Further NS5B polymerase inhibitors are currently under development. Merck is a uridine nucleo We are developing the cidoprodrug MK-3682 (formerly Idenix's IDX21437). This drug is currently in Phase II combination trials.

[0014] U.S. patent and WO applications describing nucleoside polymerase inhibitors for the treatment of flaviviridae, including HCV, include those filed by Idenix Pharmaceuticals (U.S. Patent Nos. 6,812,219, 6,914,054, 7,105,493, 7,138,376, 7,148,206, 7,157,441, 7,163,929, 7,169,766, 7,192,936, 7,365,057, 7,384,924, and 7,456). ,155, No. 7,547,704, No. 7,582,618, No. 7,608,597, No. 7,608,600, No. 7,625,875, No. 7,635,689, No. 7, 662,798, 7,824,851, 7,902,202, 7,932,240, 7,951,789, 8,193,372, 8,299,038, U.S. Patent Publication No. 8,343,937, No. 8,362,068, No. 8,507,460, No. 8,637,475, No. 8,674,085, No. 8,680,071, No. 8,691,788, No. 8,742,101, No. 8,951,985, No. 9,109,001, No. 9,243,025, U.S. Patent Application Publication No. 2016 / 0002281, U.S. Patent Application Publication No. 201 International Publication Nos. 3 / 0064794, 2015 / 095305, 2015 / 081133, 2015 / 061683, 2013 / 177219, 2013 / 039920, 2014 / 137930, 2014 / 052638, and 2012 / 154321; filed by Merck (US 6, 7 No. 77,395, No. 7,105,499, No. 7,125,855, No. 7,202,224, No. 7,323,449, No. 7 ,339,054, 7,534,767, 7,632,821, 7,879,815, 8,071,568, Publications 8,148,349, 8,470,834, 8,481,712, 8,541,434, 8,697,694, 8,715,638, 9,061,041, 9,156,872, and International Publication No. 2013 / 009737); Emory Applications filed by the University (US Patent Nos. 6,348,587, 6,911,424, 7,307,065, 7,495,006, 7,662,938, 7,772,208, 8,114,994, 8,168,583, 8,609,627, US Patent Application Publication No. 2014 / 0212382, and International Publication No. 2014 / 1244430); Applications filed by Gilead Sciences / Pharmasset Inc. (US Patent Nos. 7,842,672, 7,973,0 No. 13, No. 8,008,264, No. 8,012,941, No. 8,012,942, No. 8,318,682, No. 8,324,179, No. 8,415,308, No. 8,45 No. 5,451, No. 8,563,530, No. 8,841,275, No. 8,853,171, No. 8,871,785, No. 8,877,733, No. 8,889,159, No. 8, Nos. 906,880, 8,912,321, 8,957,045, 8,957,046, 9,045,520, 9,085,573, 9,090,642 and 9,139,604 (US Nos. 6,908,924, 6,949,522, 7,094,770, 7,211,570, 7,429,572, 7, RE No. 601,820, RE No. 7,638,502, RE No. 7,718,790, RE No. 7,772,208, RE No. 42,015, RE No. 7,919,247, RE No. 7,964,580, RE No. No. 8,093,380, No. 8,114,997, No. 8,173,621, No. 8,334,270, No. 8,415,322, No. 8,481,713, No. 8,492,539, Nos. 8,551,973, 8,580,765, 8,618,076, 8,629,263, 8,633,309, 8,642,756, 8,716,262, 8,716,263, 8,735,345, 8,735,372, 8,735,569, 8,759,510 and 8,765,710); filed by Hoffman La-Roche (US No. 6,660,721); filed by Roche (US No. 6,784,166 Patent Nos. 7,608,599, 7,608,601 and 8,071,567; filed by Alios BioPharma Inc. (U.S. Patent Nos. 8,895,723, 8,877,731, 8,871,737, 8,846,896, 8,772,474, 8,980,865, 9,012,427, U.S. Patent Application Publication No. 2015 / 0105341, U.S. Patent Application Publication No. 2015 / 0011497, U.S. Patent Application Publication No. 2010 / 0249068, U.S. Patent Application Publication International Publication No. 2012 / 0070411, International Publication No. 2015 / 054465, International Publication No. 2014 / 209979, International Publication No. 2014 / 100505, International Publication No. 2014 / 100498, International Publication No. 2013 / 142159, International Publication No. 2013 / 142157, International Publication No. 2013 / 096680, International Publication No. 2013 / 088155, International Publication No. 2010 / 108135); Enanta Applications filed by Pharmaceuticals (US Patent Nos. 8,575,119, 8,846,638, 9,085,599, International Publication No. 2013 / 044030, International Publication No. 2012 / 125900); Applications filed by Biota (US National Patent Nos. 7,268,119, 7,285,658, 7,713,941, 8,119,607, 8,415,309, 8,501,699 and 8,802,840); patents filed by Biocryst Pharmaceuticals (U.S. Patents Nos. 7,388,002, 7,429,571, 7,514,410, 7,560,434, 7,994,139, 8,133,870, 8,163,703, 8,242,085 and 8,440,813); Alla Chem, Filings filed by LLC (U.S. Patent No. 8,889,701 and International Publication No. 2015 / 053662); Filings filed by Inhibitex (U.S. Patent No. 8,759,318 and International Publication No. 2015 / 053662); Publication No. 2012 / 092484); Filings filed by Janssen Products (US Patent Nos. 8,399,429, 8,431,588, 8,481,510, 8,552,021, 8,933,052, 9,006,29 and 9,012,428); Filings filed by the University of Georgia Foundation (US Patent Nos. 6,348,587, 7,307,065, 7,662,938, 8,168,583, 8,673,926, 8,816,074, 8,921,384 and 8,946,244); Filings filed by RFS Pharma, LLC (US JAPONIE Patent Nos. 8,895,531, 8,859,595, 8,815,829, 8,609,627, and 7,560,550; U.S. Patent No. 2014 / 0066395; U.S. Patent Application Publication No. 2014 / 0235566; U.S. Patent Application Publication No. 20 10 / 0279969, International Publication No. 2010 / 091386 and International Publication No. 2012 / 158811); filed by University College Cardiff Consultants Limited (International Publication No. 2014 / 076490, International Publication No. 2010 / 081082, International Publication No. 2008 / 062206); filed by Achillion Pharmaceuticals, Inc. ( Examples include: International Publication Nos. 2014 / 169278 and 2014 / 169280; applications filed by Cocrystal Pharma, Inc. (U.S. Patent No. 9,173,893); applications filed by Katholieke Universiteit Leuven (International Publication No. 2015 / 158913); applications filed by Catabasis (International Publication No. 2013 / 090420); and applications filed by the Regents of the University of Minnesota (International Publication No. 2006 / 004637). It is possible.

[0015] Nevertheless, there is a strong medical need for the development of safe, effective, and well-tolerated anti-HCV therapies. This need is increasing due to the predicted development of drug resistance. More potent direct-acting antiviral drugs have the potential to significantly shorten the duration of treatment and improve compliance and sustained response rates (SVR) in patients infected with all HCV genotypes. [Overview of the project] [Problems that the invention aims to solve]

[0016] Therefore, it is an object of the present invention to provide compounds, pharmaceutical compositions, methods and uses for treating and / or preventing HCV infection. [Means for solving the problem]

[0017] Compounds of formulas I, II, III, IV, V, VI, and VII, and β-D-2'-deoxy-2'-α-fluoro-2'-β-C-substituted-N 6 Compounds containing (mono- or di-methyl)purine nucleotides have been found to be highly active against the HCV virus when administered in effective doses to a host requiring them. The host may be a human or any animal infected with the virus.

[0018] The disclosed nucleotides include those having nanomolar activity against HCV in vitro and a therapeutic index of 25,000 or more.

[0019] Surprisingly, the parent N of the disclosed compound 6 -(methyl)purine nucleosides had not been developed as drug candidates prior to the present invention, nor had they been specifically disclosed. For example, in 2010, 3'-azide-N 6-Dimethyl-2,6-diaminopurine has been reported to be substantially not deaminated by adenosine deaminase over a long period (120 minutes), and for this reason, it has been considered an unsuitable compound for derivatization as a drug (see, for example, page 86 of WO 2010 / 091386 and the corresponding US Patent No. 8,609,627).

[0020] However, it has now been discovered that the compounds of the present invention are assimilated into the 5-monophosphate of N 6 -substituted purines without substantial N 6 -deamination and then at the 6-position to produce active guanine triphosphate compounds such that excellent activity and therapeutic indices are obtained.

[0021] In particular, as follows, β-D-2'-deoxy-2'-α-fluoro-2'-β-methyl-N 6 -methyl-2,6-diaminopurine nucleotides, and β-D-2'-deoxy-2'-α-fluoro-2'-β-methyl-N 6 -dimethyl-2,6-diaminopurine nucleotides, and also other β-D-2'-D-2'-α-fluoro-2'-β-C-substituted-2-modified-N 6 -substituted purine nucleotide 5'-stabilized phosphate prodrugs or derivatives have been found to be very active against HCV. The activity of the parent nucleoside β-D-2'-deoxy-2'-α-fluoro-2'-β-methyl-N 50 = 15.7 micromolar) in the replicon assay indicates that the activity of β-D-2'-deoxy-2'-α-fluoro-2'-β-methyl-N 6 -methyl-2,6-diaminopurine is not suitable for use as a human drug due to insufficient activity (N 6A combination of references to International Publication No. 2010 / 091386, p. 86 and corresponding U.S. Patent No. 8,609,627 suggests that methyl-2,6-diaminopurine is not deaminated in vivo. Surprisingly, the stabilized racemic phosphate prodrug (phosphoramide) shows at least a 600-fold increase in activity in replicon assays. 50 It exhibits a value of 26 nanomoles (nM). The corresponding (S)-phosphoramide shows an increase in activity of at least 3900 times EC 50 It exhibits a 4nM concentration (see the structure below and compound 5-2 in Table 7). Therefore, it has a TC concentration exceeding 100 micromoles. 50 Therefore, the above compound has a therapeutic index of over 25,000. For comparison, sofosbuvir is EC 50 = 53 nM, TC exceeding 100 micromoles 50 , and has a treatment index of over 1920. [ka]

[0022] Similarly, replicon assay (EC 50 =10.7 micromoles, "μM") Parent nucleoside β-D-2'-deoxy-2'-α-fluoro-2'-β-methyl-N 6 The activity of -dimethyl-2,6-diaminopurine also indicates that it is unsuitable for use as a human drug due to insufficient activity, whereas the stabilized racemic phosphate prodrug (phosphoramide) shows an increase in activity of more than 890 times in replicon assays. 50 It exhibits an EC2+ of 12nM. Furthermore, the corresponding (S)-phosphorumidate (compound 25, Table 7) also shows an increase in activity of at least 2600 times. 50 It exhibits a molecular weight of 4nM (see structure below). Furthermore, compound 25 also has a therapeutic index of over 25,000. [ka]

[0023] In another example, in a replicon assay, the compound isopropyl((((R,S)-(2R,3R,4R,5R)-5-(2-amino-6-(N-methyl-N-cyclopropyl -amino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate is EC 50 It shows =7nM, and the corresponding (S)-phosphoramidate is EC2. 50 It showed a maturity of 5 nM (see Compound 27 in Table 7 and the structure below). [ka]

[0024] As described above, β-D-2'-deoxy-2'-α-fluoro-2'-β-methyl-N as a phosphoramidate 6 The metabolism of methyl-2,6-diaminopurine nucleoside involves the production of 5'-monophosphate and N-2'-deoxy-2'-α-fluoro-2'-β-methyl-guanine nucleoside as 5'-monophosphate. 6 This involves the subsequent assimilation of a methyl-2,6-diaminopurine base. The monophosphate is then further assimilated into the active species, namely the 5'-triphosphate. β-D-2'-deoxy-2'-α-fluoro-2'-β-methyl-guanine triphosphate is IC12 against HCV genotype 1b NS5B polymerase. 50 It has a concentration of 0.15 μM.

[0025] Therefore, in one embodiment, the present invention is [ka] (In the formula, Y is NR 1 R 2 And, R 1This includes C1-C5 alkyl (including methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, tert-butyl, and pentyl), C1-C5 haloalkyl (including CH2F, CHF2, CF3, CH2CF3, CF2CH3, and CF2CF3), C2-C6 alkenyl, C2-C6 alkynyl, -(C0-C2 alkyl)(C3-C6 cycloalkyl), -(C0-C2 alkyl)(heterocyclic), -(C0-C2 alkyl)(aryl), -(C0-C2 alkyl)(heteroaryl), and -OR 25 , -C(O)R 3C (-C(O)CH3, -C(O)CH2CH3, -C(O)CH(CH3)2, -C(O)OCH3, -C(O)OC2H5, -C(O)OC3 H7, -C(O)OC4H9, and -C(O)OC5H 11 (including), -C(S)R 3D , or -SO2R 28 Each of these may be substituted, R 2 This includes hydrogen, C1-C5 alkyl (including methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, tert-butyl, and pentyl), C1-C5 haloalkyl (including CHF2, CHF2, CF3, CH2CF3, and CF2CF3), -(C0-C2 alkyl)(C3-C6 cycloalkyl), and -C(O)R 3C (-C(O)CH3, -C(O)CH2CH3, -C(O)CH(CH3)2, -C(O)OCH3, -C(O)OC2H5, -C(O)OC3H7, -C(O)OC4H9, and -C(O)OC5H 11 The compounds are -(C0~C2 alkyl)(aryl), -(C0~C2 alkyl)(heterocyclic), and -(C0~C2 alkyl)(heteroaryl), where R 1 and R 2 At least one of them is methyl, CH2F, CHF2, or CF3, R 3 is hydrogen, [ka] , diphosphate, tripphosphate, substituted carbonyl-linked amino acids or -C(O)R 3C And, R 3A is, O - , may be selected from OH, -O-substituted aryls, -O-substituted heteroaryls, or substituted heterocyclines, R 3B is, O - , OH, may be selected from substituted N-linked amino acids or substituted N-linked amino acid esters, R 3C These are alkyl, alkenyl, alkynyl, -(C0~C2)(cycloalkyl), -(C0~C2)(heterocyclo), -(C0~C2)(aryl), -(C0~C2)(heteroaryl), -O-alkyl, -O-alkenyl, -O-alkynyl, -O-(C0~C2)(cycloalkyl), -O-(C0~C2)(heterocyclo), -O-(C0~C2)(aryl), or -O-(C0~C2)(heteroaryl), and each may be substituted. R 4 This is a stabilized phosphate prodrug comprising a monophosphate, diphosphate, triphophate, or, but not limited to, a phosphoramidate, thiophosphorumidate, or any other portion that is metabolized in vivo to a monophosphate, diphosphate, or triphophate in a human or animal host. Or, R 3 and R 4 Together with the oxygen to which they are bound, they can form 3',5'-cyclic prodrugs, including, but not limited to, 3',5'-cyclic phosphate prodrugs. R 12 (These are CH3, CH2F, CHF2, CF3, or ethynyl.)

[0026] In one embodiment, the present invention is [ka] (In the formula, Y is NR 1 R 2 And, R 1 This includes C1-C5 alkyl (including methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, tert-butyl, and pentyl), C1-C5 haloalkyl (including CH2F, CHF2, CF3, CH2CF3, CF2CH3, and CF2CF3), C2-C6 alkenyl, C2-C6 alkynyl, -(C0-C2 alkyl)(C3-C6 cycloalkyl), -(C0-C2 alkyl)(heterocyclic), -(C0-C2 alkyl)(aryl), -(C0-C2 alkyl)(heteroaryl), and -OR 25 , -C(O)R 3C (-C(O)CH3, -C(O)CH2CH3, -C(O)CH(CH3)2, -C(O)OCH3, -C(O)OC2H5, -C(O)OC3H7, -C(O)OC4H9 and -C(O)OC5H 11 (including), -C(S)R 3D , or -SO2R 28 Each of these may be substituted, R 2 -C(O)R 3C (-C(O)CH3, -C(O)CH2CH3, -C(O)CH(CH3)2, -C(O)OCH3, -C(O)OC2H5, -C(O)OC3H7, -C(O)OC4H9 and -C(O)OC5H 11 (including), -C(S)R 3D , or -SO2R 28 And here, R 1 and R 2At least one of them is methyl, CH2F, CHF2, or CF3, R 3 is hydrogen, [ka] , diphosphate, tripphosphate, substituted carbonyl-linked amino acids, or -C(O)R 3C And, R 3A is, O - , may be selected from OH, -O-substituted aryls, -O-substituted heteroaryls, or substituted heterocyclines, R 3B is, O - , OH, may be selected from N-linked amino acids which may be substituted or N-linked amino acid esters which may be optionally substituted, R 3C These include alkyl, alkenyl, alkynyl, -(C0~C2)(cycloalkyl), -(C0~C2)(heterocyclo), -(C0~C2)(aryl), -(C0~C2)(heteroaryl), -O-alkyl, -O-alkenyl, -O-alkynyl, -O- (C0~C2)(cycloalkyl), -O-(C0~C2)(heterocyclo), -O-(C0~C2)(aryl), -O-(C0~C2)(heteroaryl), -S-alkyl, -S-alkenyl, -S-alkynyl, -S-(C0~C2)(cycloalkyl), -S-(C0~C2)(heterocyclo), -S-(C0~C2)(aryl), or -S-(C0~C2)(heteroaryl), each of which may be substituted. R 3D These are alkyl, alkenyl, alkynyl, -(C0~C2)(cycloalkyl), -(C0~C2)(heterocyclo), -(C0~C2)(aryl), -(C0~C2)(heteroaryl), -O-alkyl, -O-alkenyl, -O-alkynyl, -O-(C0~C2)(cycloalkyl), -O-(C0~C2)(heterocyclo), -O-(C0~C2)(aryl), or -O-(C0~C2)(heteroaryl), and each may be substituted. R4 This is a stabilized phosphate prodrug comprising a monophosphate, diphosphate, triphophate, or, but not limited to, a phosphoramidate, thiophosphorumidate, or any other portion that is metabolized in vivo to a monophosphate, diphosphate, or triphophate in a human or animal host. Or, R 3 and R 4 They may, together with the oxygen to which they are bound, form 3',5'-cyclic prodrugs, including, but not limited to, 3',5'-cyclic phosphate prodrugs. R 5 This includes C1-C5 alkyl (including methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, tert-butyl and pentyl), C1-C5 haloalkyl (including CHF2, CHF2, CF3, CH2CF3 and CF2CF3), C2-C6 alkenyl, C2-C6 alkynyl, -(C0-C2 alkyl)(C3-C6 cycloalkyl), -(C0-C2 alkyl)(heterocyclic), -(C0-C2 alkyl)(aryl), -(C0-C2 alkyl)(heteroaryl), -OR 25 , -C(O)R 3C (-C(O)CH3, -C(O)CH2CH3, -C(O)CH(CH3)2, -C(O)OCH3, -C(O)OC2H5, -C(O)OC3H7, -C(O)OC4H9 and -C(O)OC5H 11 (including), -C(S)R 3D , or -SO2R 28 Each of these may be substituted, R 6is hydrogen, optionally substituted C1-C5 alkyl (including methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, tert-butyl and pentyl), C1-C5 haloalkyl (including CHF2, CHF2, CF3, CH2CF3 and CF2CF3), optionally substituted -(C0-C2 alkyl)(C3-C6 cycloalkyl), optionally substituted -(C0-C2 alkyl)(heterocyclic ring), optionally substituted -(C0-C2 alkyl)(aryl), optionally substituted -(C0-C2 alkyl)(heteroaryl), -C(O)R 3C (-C(O)CH3, -C(O)CH2CH3, -C(O)CH(CH3)2, -C(O)OCH3, -C(O)OC2H5, -C(O)OC3H7, -C(O)OC4H9 and -C(O)OC5H 11 including), -C(S)R 3D or -SO2R 28 where or R 5 and R 6 may together with the nitrogen to which they are attached form a heterocyclic ring R 12 is CH3, CH2F, CHF2, CF3 or ethynyl R 22 is Cl, Br, F, CN, N3, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, -(C1-C2 alkyl)(C3-C6 cycloalkyl), -(C0-C2 alkyl)(C3-C6 heterocyclic ring), -(C0-C2 alkyl)(aryl), -(C0-C2 alkyl)(heteroaryl), -ONHC(=O)OR 23 , -NHOR 24 , -OR 25 , -SR 25 , -NH(CH2) 1~4 N(R 26 )2, -NHNHR 26 , -N=NR 27 , -NHC(O)NHNHR 27 , -NHC(S)NHNHR 27 , -C(O)NHNHR 27 , -NR 27 SO2R 28 , -SO2NR27 R 29 -C(O)NR 27 R 29 , -CO2R 29 , -SO2R 29 , [ka] , -P(O)H(OR 29 ), -P(O)(OR 29 )(OR 30 ), -P(O)(OR 29 )(NR 29 R 30 ), or -NR 5 R 6 And, For example, but not limited to, the following embodiments: chloro, bromo, fluoro, cyano, azide, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, tert-butyl, and n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, 1-ethylpropyl, vinyl, allyl, 1-butynyl, 2-butynyl, acetylenyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, -(CH 2)-Cyclopropyl, -(CH2)-Cyclobutyl, -(CH2)-Cyclopentyl, -(CH2)-Cyclohexyl, Aziridine, Oxirane, Thiirane, Azethidine, Oxetane, Thiethane, Pyrrolidine, Tetrahydrofuran, Thiorane, Pyrazolidine, Piperidine, Oxane, Thiane, -(CH2)-Aziridine, -(CH2)-Oxirane, -(CH2)-Thiirane, -(CH2)-Azethidine, -(CH2)-Oxetane, -(CH2)-Thiethane, -(CH2)-Pyrrolidine -(CH2)-tetrahydrofuran, -(CH2)-thiolane, -(CH2)-pyrazolidine, -(CH2)-piperidine, -(CH2)-oxane, -(CH2)-thiane, phenyl, pyridyl, -ONHC(=O)OCH3, -ONHC(=O)OCH2CH3, -NHOH, NHOCH3, -OCH3, OC2H5, -OPh, OCH2Ph, -SCH3, -SC2H5, -SPh, SCH2Ph, -NH(CH2)2NH2, -NH(CH2)2N(CH3)2, -NHNH2, -NHNHCH3, -N=NH, -N=NCH3, -N=NCH2CH3, -NHC(O)NHNH2, -NHC(S)NHNH2, -C(O)NHNH2, -NHSO2CH3, -NHSO2CH2CH3, -SO2NHCH3, -SO2N( CH3)2, -C(O)NH2, -C(O)NHCH3, -C(O)N(CH3)2, -CO2CH3, -CO2CH2CH3, -CO2Ph, -CO2CH2Ph, -SO2CH3, -SO2CH2CH3, -SO2Ph, -SO2CH2Ph, [ka] , -P(O)H(OH), -P(O)H(OCH3), -P(O)(OH)(OH), -P(O)(OH)(OCH3), -P(O)(OCH3)(O CH3), -P(O)(OH)(NH2), -P(O)(OH)(NHCH3), -P(O)(OH)N(CH3)2, -NHC(O)CH3, -NHC (O)CH2CH3, -NHC(O)CH(CH3)2, -NHC(O)OCH3, -NHC(O)OCH2CH3, -NHC(O)OCH(CH3)2, -NHC(O)OCH2CH2CH3, -NHC(O)OCH2CH2CH2CH3, and -NHC(O)OCH2CH2CH2CH2CH3, R 23 These are C1-C5 alkyl, -(C0-C2 alkyl)(C3-C6 cycloalkyl), -(C0-C2 alkyl)(heterocyclic)-(C0-2 alkyl)(aryl), or -(C0-C2 alkyl)(heteroaryl), and each may be substituted. R 24 Here, hydrogen is a C1-C6 alkyl group, -(C1-C2 alkyl)(C3-C6 cycloalkyl group), -(C1-C2 alkyl)(C3-C6 heterocyclic group), -(C0-C2 alkyl)(aryl group), or -(C0-C2 alkyl)(heteroaryl group), where hydrogen may be substituted in any of the following ways: R 25 These are hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, -(C0-C2 alkyl)(C3-C6 cycloalkyl), -(C0-C2 alkyl)(C3-C6 heterocyclic), -(C0-C2 alkyl)(aryl), or -(C0-C2 alkyl)(heteroaryl), where hydrogen may be substituted in any of these cases. R 26 This is independently selected from hydrogen, C1-C6 alkyl, -(C0-C2 alkyl)(C3-C6 cycloalkyl), -(C0-C2 alkyl)(heterocyclic), -(C0-C2 alkyl)(aryl), or -(C0-C2 alkyl)(heteroaryl), where each of these may be substituted except for hydrogen. R 27 is a C1-C6 alkyl group which may be hydrogenated or substituted. R28 These are C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, -(C0-C2 alkyl)(C3-C6 cycloalkyl), -(C0-C2 alkyl)(C3-C6 heterocyclic), (C0-C2 alkyl)(aryl), or (C0-C2 alkyl)(heteroaryl), and each may be substituted. R 29 These are hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, -(C0-C2 alkyl)(C3-C6 cycloalkyl), -(C0-C2 alkyl)(C3-C6 heterocyclic), -(C0-C2 alkyl)(aryl), or -(C0-C2 alkyl)(heteroaryl), where hydrogen may be substituted in any of these cases. Or, R 27 and R 29 They may form a heterocycle with the nitrogen to which they are bonded. R 30 These are hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, -(C0-C2 alkyl)(C3-C6 cycloalkyl), -(C0-C2 alkyl)(C3-C6 heterocyclic), -(C0-C2 alkyl)(aryl), or -(C0-C2 alkyl)(heteroaryl), where hydrogen may be substituted in any of these cases. Or, R 29 and R 30 They may also combine to form a heterocycle. x is 1, 2, or 3.

[0027] β-D-2'-deoxy-2'-α-fluoro-2'-β-methyl-N 6 The metabolism of -dimethyl-2,6-diaminopurine nucleotide is β-D-2'-deoxy-2'-α-fluoro-2'-β-methyl-N 6 This involves both the formation of a dimethyl-2,6-diaminopurine nucleoside triphosphate and the generation of the corresponding guanine nucleoside triphosphate. See mechanisms 2 and 3.

[0028] 2'-Deoxy-2'-α-Fluoro-2'-β-C-Substituting-N6 -Substituted-2,6-diaminopurine nucleotides can be modified by alkylation or acylation, which can alter their lipophilicity, pharmacokinetics, and / or nucleotide targeting to the liver. 2 Further substitutions can be made at the position. N as described herein 2 -Unless the amino group is completely substituted by a different moiety such as fluoro, the 2-position of the diaminopurine is modified with 2'-deoxy-2'-α-fluoro-2'-β-C-substituted-N 6 It has been discovered that the specificity of nucleotide derivatives can be further increased both in vitro and in vivo by dealkylating or deacyling substituted-2,6-diaminopurine nucleotides with liver enzymes. For example, nucleoside phosphoramidate 2'-deoxy-2'-α-fluoro-2'-β-methyl-N 2 -methyl-N 6 -Methyl-2,6-diaminopurine nucleoside phosphoramidate, when incubated in vitro with the human liver S9 fraction for up to 60 minutes, forms 2'-deoxy-2'-α-fluoro-2'-β-methyl-N 6 -Dealkylated to methyl-2,6-diaminopurine nucleoside phosphoramidate, these conditions mimic in vivo conditions. In one embodiment, N 2 The modification increases cell permeability and hepatitis targeting.

[0029] Despite numerous antiviral nucleosides in literature and patent applications, the 2'-deoxy-2'-α-fluoro-2'-β-methyl-N described herein is not the most effective. 6 -methyl-2,6-diaminopurine nucleoside, 2'-deoxy-2'-α-fluoro-2'-β-methyl-N 6 - 5'-stabilized phosphate derivatives of dimethyl-2,6-diaminopurine nucleoside, and other β-D-2'-D-2'-α-fluoro-2'-β-C-substituted-2-modified-N 6 -Substituted purine nucleoside derivatives are not specifically disclosed, nor are their advantageous activities described.

[0030] Unless otherwise specified, the compounds described herein are provided in the β-D configuration. Similarly, in the form of phosphoramides or thiophosphoramides, the amino acid moiety may be in the L or D configuration. In alternative embodiments, the above compounds may be provided in the β-L configuration. Similarly, any substituent exhibiting chirality may be provided in racemic, enantiomer, diastereomer, or any mixture thereof. Phosphoramides, thiophosphoramides, or other stabilized phosphorus prodrugs exhibiting chirality are R 4 When used as a stabilized phosphate prodrug, the phosphorus prodrug can be provided as a racemic mixture of R or S chiral phosphorus derivatives, or a mixture thereof. All of these combinations of stereochemistry are included in the invention described herein.

[0031] Accordingly, the present invention comprises compounds of formulas I to VII as described herein, or pharmaceutically acceptable compositions, salts, or prodrugs thereof. [ka]

[0032] In one particular embodiment, the parent nucleoside, namely R 4 Nucleosides having hydrogen and therefore a hydroxyl group at the 5' position are not deaminated by adenosine deaminase for 7 minutes, 10 minutes, 30 minutes, 60 minutes, or 120 minutes under conditions that substantially mimic the in vivo environment (e.g., ambient temperature and aqueous physiological pH). Unless otherwise stated, the time is 30 minutes. In this embodiment, the term "substantially deaminated" means that the parent compound is not converted to the corresponding guanine derivative, i.e., the 6-oxo derivative, in an amount sufficient to provide a therapeutic effect in vivo.

[0033] Compounds, methods, and compositions are provided for the treatment of a host infected with the HCV virus by administration of an effective amount of the compound or a pharmaceutically acceptable salt thereof.

[0034] Furthermore, the compounds and compositions can be used to treat related conditions such as anti-HCV antibody-positive and antigen-positive states, chronic liver inflammation caused by the virus, liver cancer resulting from advanced hepatitis C, cirrhosis, chronic or acute hepatitis C, fulminant hepatitis C, chronic persistent hepatitis C, and fatigue caused by anti-HCV. Additionally, in individuals who are anti-HCV antibody or antigen-positive, or who have been exposed to hepatitis C, the compounds and compositions can be used to clinically prevent the disease or suppress its progression. The compound or a formulation containing the compound may be used preventively.

[0035] In another embodiment, formula Ia: [ka] (In the formula, Y, R 3 and R 4 We disclose compounds of (as defined above).

[0036] In one embodiment of formula Ia, R 3 It is hydrogen.

[0037] In one embodiment of formula Ia, Y is NR 1 R 2 If R 1 is methyl, and R 2 It is hydrogen.

[0038] In one embodiment of formula Ia, Y is NR 1 R 2 If R 1 and R 2 All of them are methyl.

[0039] In one embodiment of formula Ia, Y is NR 1 R 2 If R 1 is methyl, and R 2 It is cyclopropyl.

[0040] In another embodiment, formula Ib: [ka] (In the formula, Y, R 3 and R 4 We disclose compounds of (as defined above).

[0041] In one embodiment of formula Ib, R 3 It is hydrogen.

[0042] In one embodiment of formula Ib, Y is NR 1 R 2 If R 1 is methyl, and R 2 It is hydrogen.

[0043] In one embodiment of formula Ib, Y is NR 1 R 2 If R 1 and R 2 All of them are methyl.

[0044] In one embodiment, formula II: [ka] (In the formula, Y, R 3 , R 4 , R 12 and R 22 We disclose compounds of (as defined above).

[0045] In another embodiment, formula IIa: [ka] (In the formula, Y, R 3 , R 4 and R 22 We disclose compounds of (as defined above).

[0046] In another embodiment, formula IIb: [ka] (In the formula, Y, R 3 , R 4 and R 22 We disclose compounds of (as defined above).

[0047] In one embodiment, Equation III: [ka] (In the formula, variables Y and R 3 , R 7 , R 8 , R 9a , R 9b , R 10 , R 12 and R 22 This specification discloses compounds of the type described herein.

[0048] In one embodiment, formula IV: [ka] (In the formula, variables Y and R 3 , R 7 , R 8 , R 9a , R 9b , R 10 and R 22 This specification discloses compounds of the type described herein.

[0049] In one embodiment, formula V: [ka] (In the formula, variables Y and R 3 , R 7 , R 8 , R 9a , R 9b , R 10 and R 22 This specification The compounds described are disclosed.

[0050] In one embodiment, formula VI: [ka] (In the formula, R 41 This includes halogens (especially F or Cl), OR 3 , N3, NH2 or CN, and also variables Y, R 3 , R 4 and R 12 This specification discloses compounds of the type described herein.

[0051] In one embodiment, formula VII: [ka] (In the formula, variables Y and R 3 , R 4 , R 12 and R 41 This specification discloses compounds of the type described herein.

[0052] The phosphorus in any of the above formulas may be chiral and therefore may be provided as an enantiomer of R or S, or a mixture thereof including a racemic mixture.

[0053] Compound 5 was separated into enantiomers 5-1 and 5-2. Compound 5-2 was also prepared by chiral synthesis and designated as compound 24.

[0054] In one embodiment, compounds, methods, and compositions for the treatment of a host infected with or exposed to hepatitis C are provided herein. The compounds of the present invention can be administered alone or in combination with another anti-HCV agent in an amount effective to treat the infected host. In certain embodiments, it is useful to administer a combination of drugs that modulate the same or different pathways or inhibit different targets in the virus. Disclosed β-D-2'-D-2'-α-fluoro-2'-β-C-substituted-2-modified-N 6 - Substituting purine nucleotides are NS5B polymerase inhibitors, therefore NS3 / 4A protea It may be useful to administer the compound to the host in combination with a protease inhibitor such as an NS5A inhibitor (e.g., telaprevir (Insivec®), boceprevir (Victorelis®), simeprevir (Olysio®), or paritaprevir), or an NS5A inhibitor (e.g., ombitasvir). Furthermore, the compound of the present invention can be administered in combination with a structurally different NS5B polymerase inhibitor, such as another compound described herein or below, including Gilead's Sovaldi®. Additionally, the compound of the present invention can be administered in combination with interferon alpha-2a, which may be pegylated or otherwise modified, and / or ribavirin.

[0055] The present invention β-D-2'-D-2'-α-fluoro-2'-β-C-substitution-2-modification-N 6 - Substituted purine nucleotides are typically administered orally, for example, in the form of pills or tablets, but may also be administered by other routes as appropriate by the attending physician, including intravenous, percutaneous, subcutaneous, topical, parenteral, or other suitable routes. [Brief explanation of the drawing]

[0056] [Figure 1] This is a semi-prep run chromatogram illustrating the separation of stereoisomers of compound 5 using the Phenominex Luna column disclosed in Example 9. The y-axis is shown in units of mAU, and the x-axis is measured in minutes. [Figure 2] This graph shows the HCV replication inhibition curves for compound 5-2 (Table 7) and sofosbuvir. Compound 5-2 has an EC50 of 4 nM, a TC50 of over 100 micromoles, and a therapeutic index of over 25,000. Sofosbuvir has an EC50 of 53 nM, a TC50 of over 100 micromoles, and a therapeutic index of over 1,920. The y-axis represents the percentage relative to the viral control, and the x-axis represents the drug concentration in μM units. [Figure 3]This graph shows the HCV replication inhibition curves for compound 25 (Table 7) and sofosbuvir. As described in Example 27, compound 25 has an EC50 of 4 nM, a TC50 greater than 100 μM, and a therapeutic index greater than 25,000. Sofosbuvir has an EC50 of 53 nM, a TC50 greater than 100 micromoles, and a therapeutic index greater than 1,920. The y-axis represents the percentage relative to the viral control, and the x-axis represents the drug concentration in μM. [Figure 4] This figure shows the intraassay comparison of anti-HCV activity for compounds 5-2, 25, 27 (Table 7), and sofosbuvir. The y-axis represents the percentage relative to the viral control, and the x-axis represents the drug concentration in μM. See Example 27. [Figure 5] This graph shows the stability of compound 5-2, compound 5-2 N2-acetate, compound 5-2 N2-butyrate, compound 5-2 N2-methyl derivative, and compound 5-2 N2-n-pentylcarbamate in human blood. The x-axis represents incubation time measured in minutes, and the y-axis represents the percentage of residual parent compound. [Figure 6] This graph shows the in vitro time-dependent dealkylation of 2'-deoxy-2'-α-fluoro-2'-β-methyl-N2-methyl-N6-methyl-2,6-diaminopurine nucleoside phosphoramide to 2'-deoxy-2'-α-fluoro-2'-β-methyl-N6-methyl-2,6-diaminopurine nucleoside phosphoramide in the presence of human liver S9 fraction. The x-axis represents measurements in minutes, and the y-axis represents the measured concentration of the residual compound in nM. [Figure 7] This graph shows the stability of compound 5-2, compound 5-2 N2-acetate, compound 5-2 N2-butyrate, compound 5-2 N2-methyl derivative, and compound 5-2 N2-n-pentylcarbamate in the presence of human liver S9 fraction. The x-axis represents measurements in minutes, and the y-axis represents the measured percentage of residual compound. [Figure 8]This figure shows the metabolites of 25 major compounds produced in human liver cells. The x-axis represents incubation time in hours. The y-axis represents intracellular concentration in pmol / 10⁶ cells. See Example 33. [Figure 9] This figure shows the metabolites of 27 major compounds produced in human liver cells. The x-axis represents incubation time in hours. The y-axis represents intracellular concentration in pmol / 10⁶ cells. See Example 33. [Figure 10] This figure shows the metabolites of the main compound 5-2 produced in human liver cells. The x-axis represents incubation time in hours. The y-axis represents intracellular concentration in pmol / 10⁶ cells. See Example 33. [Figure 11] This graph shows the activation pathways for compounds 25, 27, and 5-2. As can be seen from the figure, compounds 25, 27, and 5-2 are converted to their corresponding monophosphate analogs and then metabolized to a common MP analog, namely β-D-2'-deoxy-2'-α-fluoro-2'-β-methyl-guanine monophosphate. The monophosphate is then stepwise phosphorylated to the active triphosphate, namely β-D-2'-deoxy-2'-α-fluoro-2'-β-methyl-guanine triphosphate. See Example 33. [Modes for carrying out the invention]

[0057] The inventions disclosed herein are compounds, methods, and compositions for the treatment of infection with or exposure to the HCV virus in humans and other host animals, comprising the administration of an effective amount of a compound of formulas I to VII described herein or a pharmaceutically acceptable salt or prodrug thereof in an optionally pharmaceutically acceptable carrier. The compounds of the present invention are either antiviral in nature or metabolized into compounds exhibiting such activity.

[0058] Furthermore, the above compounds and compositions can be used to treat medical conditions associated with or resulting from HCV virus exposure. For example, active compounds can be used to treat HCV antibody-positive and HCV antigen-positive conditions, chronic liver inflammation caused by the virus, liver cancer resulting from severe hepatitis C, cirrhosis, acute hepatitis C, fulminant hepatitis C, chronic persistent hepatitis C, and anti-HCV fatigue. In one embodiment, the compound or a formulation containing the compound can also be used prophylactically to prevent or slow the progression of clinical disease in individuals who are HCV antibody or HCV antigen positive or who have been exposed to hepatitis C.

[0059] In particular, as shown below, β-D-2'-deoxy-2'-α-fluoro-2'-β-methyl-N 6 -methyl-2,6-diaminopurine nucleotide, and β-D-2'-deoxy-2'-α-fluoro-2'-β-methyl-N 6 -dimethyl-2,6-diaminopurine nucleotide, and other β-D-2'-D-2'-α-fluoro-2'-β-C-substituted-2-modified-N 6 - 5'-stabilized phosphate prodrugs or derivatives of substituted purine nucleotides were found to be highly active against HCV. The parent nucleoside in the replicon assay was β-D-2'-deoxy-2'-α-fluoro-2'-β-methyl-N 6 -Activity of methyl-2,6-diaminopurine (EC) 50 While EC2 (15.7 micromoles) indicates insufficient activity and unsuitability for use as a human drug, stabilized phosphate prodrugs (phosphoramides) show at least an 870-fold increase in activity in replicon assays. 50 This is surprising, given that it shows 26 nanomoles. Similarly, the parent nucleoside β-D-2'-deoxy-2'-α-fluoro-2'-β-methyl-N in the replicon assay. 6 Activity of dimethyl-2,6-diaminopurine (EC2) 50While a value of 10.7 micromoles (μM) indicates that it is unsuitable for use as a human drug due to insufficient activity, stabilized phosphate prodrugs (phosphoramides) show an increase in activity of more than 1300 times in replicon assays. 50 = 12 nanomoles ("nM").

[0060] Despite numerous antiviral nucleoside literature and patent applications, 2'-deoxy-2'-α-fluoro-2'-β-methyl-N 6 -methyl-2,6-diaminopurine nucleotide Reotide, 2'-deoxy-2'-α-fluoro-2'-β-methyl-N 6 -dimethyl-2,6-diaminopurine nucleotide, and other β-D-2'-D-2'-α-fluoro-2'-β-C-substituted-2-modified-N 6 -Specific 5'-stabilized phosphate derivatives of substituted purine nucleotides are not disclosed.

[0061] Unless otherwise specified, the compounds described herein are provided in the β-D configuration. In alternative embodiments, the compounds may be provided in the β-L configuration. Similarly, any substituent exhibiting chirality may be provided in racemic, enantiomer, diastereomer, or any mixture thereof. Phosphorus is a phosphoramidate, thiophosphoramide, or other stabilized phosphorus prodrug exhibiting chirality. 4 When used as a stabilized phosphate prodrug, it may be provided as a racemic mixture of R or S chiral phosphorus derivatives, or a mixture thereof. The amino acids of the phosphoramidate or thiophosphorumidate may be in a racemic mixture of D or L configurations, or a mixture thereof. Any combination of these stereoconfigurations is included in the invention described herein.

[0062] This invention includes the following features. (a) Compounds of formulas I to VII as described herein, as well as pharmaceutically acceptable salts and prodrugs thereof, (b) Formulas I to VII described herein, and their pharmaceutically acceptable salts and prodrugs, for use in the treatment or prevention of hepatitis C virus infection. (c) Use of formulas I to VII, and their pharmaceutically acceptable salts and prodrugs, in the manufacture of pharmaceuticals for the treatment of hepatitis C virus infection. (d) A method for manufacturing a pharmaceutical product intended for therapeutic use to treat hepatitis C virus infection, characterized in that formulas I to VII described herein are used in the manufacturing process. (e) Pharmaceutical preparations comprising, together with a pharmaceutically acceptable carrier or diluent, an amount effective to treat a host of formulas I to VII, or a pharmaceutically acceptable salt or prodrug thereof; (f) Formulas I to VII described herein, in substantially the absence of stereoisomers of the compounds described herein, or substantially separated from other chemical components, (g) A method for preparing a therapeutic product containing formulas I to VII described herein in an effective amount.

[0063] I. The present invention of 2'-deoxy-2'-α-fluoro-2'-β-C-substituted-2-modified-N 6 - Substituted purine nucleotides The active compound of the present invention may be provided as a pharmaceutically acceptable composition, salt, or prodrug, for example, represented by formula I: [ka] (In the formula, Y is NR 1 R 2 And, R 1 This includes C1-C5 alkyl groups (methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, tert-butyl, and pentyl), C1-C5 haloalkyl (including CH2F, CH2F, CF3, CH2CF3, CF2CH3, and CF2CF3), C2-C6 alkenyl, C2-C6 alkynyl, -(C0-C2 alkyl)(C3-C6 cycloalkyl), -(C0-C2 alkyl)(heterocyclic), -(C0-C2 alkyl)(aryl), -(C0-C2 alkyl)(heteroaryl), -OR 25 , -C(O)R 3C (-C(O)CH3, -C(O)CH2CH3, -C(O)CH(CH3)2, -C(O)OCH3, -C(O)OC2H5, -C(O)OC3H7, -C(O)OC4H9 and -C(O)OC5H 11 (including), -C(S)R 3D , or -SO2R 28 Each of these may be substituted, R 2 -C(O)R 3C (-C(O)CH3, -C(O)CH2CH3, -C(O)CH(CH3)2, -C(O)OCH3, -C(O)OC2H5, -C(O)OC3H7, -C(O)OC4H9 and -C(O)OC5H 11 (including), -C(S)R 3D or -SO2R 28 And here, R 1 and R 2 At least one of them is methyl, CH2F, CHF2, or CF3, R 3 is hydrogen, [ka] , diphosphate, tripphosphate, substituted carbonyl-linked amino acids, or -C(O)R 3C And, R 3A is, O - , may be selected from OH, -O-substituted aryls, -O-substituted heteroaryls, or substituted heterocyclines, R 3B is, O - , OH, may be selected from substituted N-linked amino acids, or substituted N-linked amino acid esters, R 3C These are alkyl, alkenyl, alkynyl, -(C0~C2)(cycloalkyl), -(C0~C2)(heterocyclo), -(C0~C2)(aryl), -(C0~C2)(heteroaryl), -O-alkyl, -O-alkenyl, -O-alkynyl, -O-(C0~C2)(cycloalkyl), -O-(C0~C2)(heterocyclo), -O-(C0~C2)(aryl), or -O-(C0~C2)(heteroaryl), and each may be substituted. R 4 This is a stabilized phosphate prodrug comprising a monophosphate, diphosphate, triphophate, or, but not limited to, a phosphoramidate, thiophosphorumidate, or any other portion that is metabolized in vivo to a monophosphate, diphosphate, or triphophate in a human or animal host. Or, R 3 and R 4 They may form 3',5'-cyclic prodrugs, including, but not limited to, 3',5'-cyclic phosphate prodrugs, together with the oxygen to which they are bound. R 12 (These are CH3, CH2F, CHF2, CF3, or ethynyl).

[0064] A stabilized phosphate prodrug is any portion capable of delivering a monophosphate, diphosphate, or triphot phosphate.

[0065] In another embodiment, formula Ia: [ka] (In the formula, Y, R 3 and R 4 We disclose compounds of (as defined above).

[0066] In another embodiment, formula Ib: [ka] (In the formula, Y, R 3 and R 4 We disclose compounds of (as defined above).

[0067] In another embodiment, the above compound is formulated with formula Ic: [ka] (In the formula, R 7 is hydrogen, C 1~6 Alkyl, C 3~7 A cycloalkyl, heteroaryl, heterocyclic, or, but not limited to, an aryl group containing phenyl or naphthyl, where phenyl or naphthyl is C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Alkoxy, F, Cl, Br, I, Nitro, Cyano, C 1~6 Haloalkyl, -N(R) 7’ )2, C 1~6 Acylamino, NHSO2C 1~6 Alkyl, -SO2N(R 7’ )2, COR 7’’ , and -SO2C 1~6 It may also be substituted with alkyl (R 7’ These are, independently, hydrogen or C 1~6 It is alkyl, R 7’’ は-OR 11 or N(R 7 )2) R 8is hydrogen, C 1~6 Alkyl, or R 9a Or R 9b and R 8 (CH2) n Here, n is 2 to 4, R 9a and R 9b teeth, (i) Hydrogen, C 1~6 Alkyl, cycloalkyl, -(CH2) c (NR 9’ )2, C 1~6 Hydroxyalkyl, --CH2SH, -(CH2)2S(O)(Me, -(CH2)3NHC(=NH)NH2, (1H-indole-3-yl)methyl, (1H-imidazole-4-yl)methyl, -(CH2) c COR 9’’ , aryl and aryl (C 1~3 Independently selected from alkyl)-, the aryl group is hydroxyl, C 1~6 Alkyl, C 1~6 They may be substituted with groups selected from alkoxy, halogen, nitro, and cyano. (ii)R 9a and R 9b All are C 1~6 It is alkyl, (iii)R 9a and R 9b (CH2) r And, (iv)R 9a is hydrogen, R 9b and R 8 Together, they form a cyclic ring containing adjacent N and C atoms (CH2) n And, (v)R 9b is hydrogen, R 9a and R 8 Together, they form a cyclic ring containing adjacent N and C atoms (CH2) n Here, c is 1 to 6, n is 2 to 4, r is 2 to 5, and R 9’ These are independently hydrogen or C 1~6It is alkyl, R 9’’ は-OR 11 or -N(R 11’ )2) and (vi)R 9a is hydrogen, R 9b These are hydrogen, CH3, CH2CH3, CH(CH3)2, CH2CH(CH3)2, CH(CH3)CH2CH3, CH2Ph, CH2-indole-3-yl, -CH2CH2SCH3, CH2CO2H, CH2C(O)NH2, CH2CH2COOH, CH2CH2C(O)NH2, CH2CH2CH2CH2NH2, -CH2CH2CH2NHC(NH)NH2, CH2-imidazole-4-yl, CH2OH, CH(OH)CH3, CH2((4'-OH)-Ph), CH2SH, or a lower cycloalkyl group. Alternatively, (vii)R 9a These are CH3, CH2CH3, CH(CH3)2, CH2CH(CH3)2, CH(CH3)CH2CH3, CH2Ph, CH2-indole-3-yl, -CH2CH2SCH3, CH2CO2H, CH2C(O)NH2, CH2CH2COOH, CH2CH2C(O)NH2, CH2CH2CH2CH2NH2, -CH2CH2CH2NHC(NH)NH2, CH2-imidazole-4-yl, CH2OH, CH(OH)CH3, CH2((4'-OH)Ph), CH2SH, or lower cycloalkyl, R 9b It is hydrogen, R 10 C may be substituted with hydrogen, alkoxy, di(lower alkyl)-amino, or halogen. 1~6 Alkyl, C 1~6 Haloalkyl, C 3~7 These include cycloalkyl, heterocycloalkyl, aminoacyl, aryl such as phenyl, heteroaryl such as pyridinyl, substituted aryl, or substituted heteroaryl. R 11 C may be substituted. 1~6 alkyl, possibly substituted cycloalkyl, possibly substituted C 2~6 Alkinyl substitution, KidC 2~6 Alkenyl or substituted acyl, but not limited to C(O)(C1~6 Contains alkyl, Y, R 3 and R 12 This is by means of (as defined herein).

[0068] In one embodiment, Formula II: [ka] (In the formula, Y is NR 1 R 2 And, R 1 This includes C1-C5 alkyl (including methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, tert-butyl, and pentyl), C1-C5 haloalkyl (including CH2F, CHF2, CF3, CH2CF3, CF2CH3, and CF2CF3), C2-C6 alkenyl, C2-C6 alkynyl, -(C0-C2 alkyl)(C3-C6 cycloalkyl), -(C0-C2 alkyl)(heterocyclic), -(C0-C2 alkyl)(aryl), -(C0-C2 alkyl)(heteroaryl), and -OR 25 , -C(O)R 3C (-C(O)CH3, -C(O)CH2CH3, -C(O)CH(CH3)2, -C(O)OCH3, -C(O)OC2H5, -C(O)OC3H7, -C(O)OC4H9 and -C(O)OC5H 11 (including), -C(S)R 3D , or -SO2R 28 Each of these may be substituted, R 2-C(O)R 3C (-C(O)CH3, -C(O)CH2CH3, -C(O)CH(CH3)2, -C(O)OCH3, -C(O)OC2H5, -C(O)OC3H7, -C(O)OC4H9 and -C(O)OC5H 11 (including), -C(S)R 3D , or -SO2R 28 And here, R 1 and R 2 At least one of them is methyl, CH2F, CHF2, or CF3, R 3 is hydrogen, [ka] , diphosphate, tripphosphate, substituted carbonyl-linked amino acids, or -C(O)R 3C And, R 3A is, O - , may be selected from OH, -O-substituted aryls, -O-substituted heteroaryls, or substituted heterocyclines, R 3B is, O - , OH, may be selected from N-linked amino acids which may be substituted or N-linked amino acid esters which may be optionally substituted, R 3CThese include alkyl, alkenyl, alkynyl, -(C0~C2)(cycloalkyl), -(C0~C2)(heterocyclo), -(C0~C2)(aryl), -(C0~C2)(heteroaryl), -O-alkyl, -O-alkenyl, -O-alkynyl, -O- (C0~C2)(cycloalkyl), -O-(C0~C2)(heterocyclo), -O-(C0~C2)(aryl), -O-(C0~C2)(heteroaryl), -S-alkyl, -S-alkenyl, -S-alkynyl, -S-(C0~C2)(cycloalkyl), -S-(C0~C2)(heterocyclo), -S-(C0~C2)(aryl), or -S-(C0~C2)(heteroaryl), each of which may be substituted. R 3D These are alkyl, alkenyl, alkynyl, -(C0~C2)(cycloalkyl), -(C0~C2)(heterocyclo), -(C0~C2)(aryl), -(C0~C2)(heteroaryl), -O-alkyl, -O-alkenyl, -O-alkynyl, -O-(C0~C2)(cycloalkyl), -O-(C0~C2)(heterocyclo), -O-(C0~C2)(aryl), or -O-(C0~C2)(heteroaryl), and each may be substituted. R 4 This is a stabilized phosphate prodrug comprising a monophosphate, diphosphate, triphophate, or, but not limited to, a phosphoramidate, thiophosphorumidate, or any other portion that is metabolized in vivo to a monophosphate, diphosphate, or triphophate in a human or animal host. Or, R 3 and R 4 They may, together with the oxygen to which they are bound, form 3',5'-cyclic prodrugs, including, but not limited to, 3',5'-cyclic phosphate prodrugs. R 5This includes C1-C5 alkyl (including methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, tert-butyl and pentyl), C1-C5 haloalkyl (including CHF2, CHF2, CF3, CH2CF3 and CF2CF3), C2-C6 alkenyl, C2-C6 alkynyl, -(C0-C2 alkyl)(C3-C6 cycloalkyl), -(C0-C2 alkyl)(heterocyclic), -(C0-C2 alkyl)(aryl), -(C0-C2 alkyl)(heteroaryl), -OR 25 , -C(O)R 3C (-C(O)CH3, -C(O)CH2CH3, -C(O)CH(CH3)2, -C(O)OCH3, -C(O)OC2H5, -C(O)OC3H7, -C(O)OC4H9 and -C(O)OC5H 11 (including), -C(S)R 3D , or -SO2R 28 Each of these may be substituted, R 6 -C(O)R 3C (-C(O)CH3, -C(O)CH2CH3, -C(O)CH(CH3)2, -C(O)OCH3, -C(O)OC2H5, -C(O)OC3H7, -C(O)OC4H9 and -C(O)OC5H 11 (including), -C(S)R 3D , or -SO2R 28 And, Or, R 5 and R 6 They may form a heterocycle with the nitrogen to which they are bonded. R 12These are CH3, CH2F, CHF2, CF3, or ethynyl. R 22 Cl, Br, F, CN, N3, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, -(C1-C2 alkyl)(C3-C6 cycloalkyl), -(C0-C2 alkyl)(C3-C6 heterocyclic), -(C0-C2 alkyl)(aryl), -(C0-C2 alkyl)(heteroaryl), -ONHC(=O)OR 23 , -NHOR 24 , -OR 25 , -SR 25 , -NH(CH2) 1~4 N(R 26 )2, -NHNHR 26 -N=NR 27 ,-NHC(O)NHNHR 27 ,-NHC(S)NHNHR 27 -C(O)NHNHR 27 , -NR 27 SO2R 28 -SO2NR 27 R 29 -C(O)NR 27 R 29 , -CO2R 29 , -SO2R 29 , [ka] , -P(O)H(OR 29 ), -P(O)(OR 29 )(OR 30 ), -P(O)(OR 29 )(NR 29 R 30 ), or -NR 5 R 6 And, For example, but not limited to, the following embodiments: chloro, bromo, fluoro, cyano, azide, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, tert-butyl, and n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, 1-ethylpropyl, vinyl, allyl, 1-butynyl, 2-butynyl, acetylenyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, -(CH 2)-Cyclopropyl, -(CH2)-Cyclobutyl, -(CH2)-Cyclopentyl, -(CH2)-Cyclohexyl, Aziridine, Oxirane, Thiirane, Azethidine, Oxetane, Thiethane, Pyrrolidine, Tetrahydrofuran, Thiorane, Pyrazolidine, Piperidine, Oxane, Thiane, -(CH2)-Aziridine, -(CH2)-Oxirane, -(CH2)-Thiirane, -(CH2)-Azethidine, -(CH2)-Oxetane, -(CH2)-Thiethane, -(CH2)-Pyrrolidine -(CH2)-tetrahydrofuran, -(CH2)-thiolane, -(CH2)-pyrazolidine, -(CH2)-piperidine, -(CH2)-oxane, -(CH2)-thiane, phenyl, pyridyl, -ONHC(=O)OCH3, -ONHC(=O)OCH2CH3, -NHOH, NHOCH3, -OCH3, OC2H5, -OPh, OCH2Ph, -SCH3, -SC2H5, -SPh, SCH2Ph, -NH(CH2)2NH2, -NH(CH2)2N(CH3)2, -NHNH2, -NHNHCH3, -N=NH, -N=NCH3, -N=NCH2CH3, -NHC(O)NHNH2, -NHC(S)NHNH2, -C(O)NHNH2, -NHSO2CH3, -NHSO2CH2CH3, -SO2NHCH3, -SO2N( CH3)2, -C(O)NH2, -C(O)NHCH3, -C(O)N(CH3)2, -CO2CH3, -CO2CH2CH3, -CO2Ph, -CO2CH2Ph, -SO2CH3, -SO2CH2CH3, -SO2Ph, -SO2CH2Ph, [ka] , -P(O)H(OH), -P(O)H(OCH3), -P(O)(OH)(OH), -P(O)(OH)(OCH3), -P(O)(OCH3)(O CH3), -P(O)(OH)(NH2), -P(O)(OH)(NHCH3), -P(O)(OH)N(CH3)2, -NHC(O)CH3, -NHC (O)CH2CH3, -NHC(O)CH(CH3)2, -NHC(O)OCH3, -NHC(O)OCH2CH3, -NHC(O)OCH(CH3)2, -NHC(O)OCH2CH2CH3, -NHC(O)OCH2CH2CH2CH3, and -NHC(O)OCH2CH2CH2CH2CH3, R 23 These are C1-C5 alkyl, -(C0-C2 alkyl)(C3-C6 cycloalkyl), -(C0-C2 alkyl)(heterocyclic), -(C0-2 alkyl)(aryl), or -(C0-C2 alkyl)(heteroaryl), and each may be substituted. R 24 Here, hydrogen is a C1-C6 alkyl group, -(C1-C2 alkyl)(C3-C6 cycloalkyl group), -(C1-C2 alkyl)(C3-C6 heterocyclic group), -(C0-C2 alkyl)(aryl group), or -(C0-C2 alkyl)(heteroaryl group), where hydrogen may be substituted in any of the following ways: R 25 These are hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, -(C0-C2 alkyl)(C3-C6 cycloalkyl), -(C0-C2 alkyl)(C3-C6 heterocyclic), -(C0-C2 alkyl)(aryl), or -(C0-C2 alkyl)(heteroaryl), where hydrogen may be substituted in any of these cases. R 26 This is independently selected from hydrogen, C1-C6 alkyl, -(C0-C2 alkyl)(C3-C6 cycloalkyl), -(C0-C2 alkyl)(heterocyclic), -(C0-C2 alkyl)(aryl), or -(C0-C2 alkyl)(heteroaryl), where each of these may be substituted except for hydrogen. R 27 is a C1-C6 alkyl group which may be hydrogenated or substituted. R28 These are C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, -(C0-C2 alkyl)(C3-C6 cycloalkyl), -(C0-C2 alkyl)(C3-C6 heterocyclic), -(C0-C2 alkyl)(aryl), or -(C0-C2 alkyl)(heteroaryl), and each may be substituted. R 29 These are hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, -(C0-C2 alkyl)(C3-C6 cycloalkyl), -(C0-C2 alkyl)(C3-C6 heterocyclic), -(C0-C2 alkyl)(aryl), or -(C0-C2 alkyl)(heteroaryl), where hydrogen may be substituted in any of these cases. Or, R 27 and R 29 They may form a heterocycle with the nitrogen to which they are bonded. R 30 These are hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, -(C0-C2 alkyl)(C3-C6 cycloalkyl), -(C0-C2 alkyl)(C3-C6 heterocyclic), -(C0-C2 alkyl)(aryl), or -(C0-C2 alkyl)(heteroaryl), where hydrogen may be substituted in any of these cases. Or, R 29 and R 30 They may also combine to form a heterocycle. The present invention discloses a compound of x (where x is 1, 2, or 3).

[0069] In another embodiment, formula IIa: [ka] (In the formula, Y, R 3 , R 4 and R 22 We disclose compounds of (as defined above).

[0070] In another embodiment, formula IIb: [ka] (In the formula, Y, R 3 , R 4 and R 22 We disclose compounds of (as defined above).

[0071] In a typical embodiment, the compound is a β-D isomer (i.e., the naturally occurring configuration) with respect to the corresponding nucleoside. In an alternative configuration, the compound is provided as a β-L isomer. The compound is typically free of at least 90% of the opposite enantiomer, and may be free of at least 98%, 99%, or even 100% of the opposite enantiomer. Unless otherwise stated, the compound is free of at least 90% of the opposite enantiomer.

[0072] In another embodiment, the above compound is of formula III: [ka] (In the formula, R 7 is hydrogen, C 1~6 Alkyl, C 3~7 A cycloalkyl, heteroaryl, heterocyclic group, or, but not limited to, an aryl group containing phenyl or naphthyl, where phenyl or naphthyl is C 1~6 Alkyl, C 2~6 Alkenil, C 2~6 Alkinyl, C 1~6 Alkoxy, F, Cl, Br, I, Nitro, Cyano, C 1~6 Haloalkyl, -N(R) 7’ )2, C 1~6 Acylamino, NHSO2C 1~6 Alkyl, -SO2N(R 7’ )2, COR 7’’ , and -SO2C 1~6 It may also be substituted with alkyl (R 7’ These are, independently, hydrogen or C 1~6 It is alkyl, R 7’’ は-OR 11 or N(R 7 )2) R 8 is hydrogen, C 1~6Alkyl, or R 9a Or R 9b and R 8 (CH2) n Here, n is 2 to 4, R 9a and R 9b teeth, (i) Hydrogen, C 1~6 Alkyl, cycloalkyl, -(CH2) c (NR 9’ )2, C1 ~6 Hydroxyalkyl, --CH2SH, -(CH2)2S(O)(Me, -(CH2)3NHC(=NH)NH2, (1H-indole-3-yl)methyl, (1H-imidazole-4-yl)methyl, -(CH2) c COR 9’’ , aryl and aryl (C 1~3 Independently selected from alkyl)-, the aryl group is hydroxyl, C 1~6 Alkyl, C 1~6 They may be substituted with groups selected from alkoxy, halogen, nitro, and cyano. (ii)R 9a and R 9b All are C 1~6 It is alkyl, (iii)R 9a and R 9b (CH2) r And, (iv)R 9a is hydrogen, R 9b and R 8 Together, they form a cyclic ring containing adjacent N and C atoms (CH2) n And, (v)R 9b is hydrogen, R 9a and R 8 Together, they form a cyclic ring containing adjacent N and C atoms (CH2) n Here, c is 1 to 6, n is 2 to 4, r is 2 to 5, and R 9’ These are independently hydrogen or C 1~6It is alkyl, R 9’’ は-OR 11 or -N(R 11’ )2) and (vi)R 9a is hydrogen, R 9b These are hydrogen, CH3, CH2CH3, CH(CH3)2, CH2CH(CH3)2, CH(CH3)CH2CH3, CH2Ph, CH2-indole-3-yl, -CH2CH2SCH3, CH2CO2H, CH2C(O)NH2, CH2CH2COOH, CH2CH2C(O)NH2, CH2CH2CH2CH2NH2, -CH2CH2CH2NHC(NH)NH2, CH2-imidazole-4-yl, CH2OH, CH(OH)CH3, CH2((4'-OH)-Ph), CH2SH, or a lower cycloalkyl group. Alternatively, (vii)R 9a These are CH3, CH2CH3, CH(CH3)2, CH2CH(CH3)2, CH(CH3)CH2CH3, CH2Ph, CH2-indole-3-yl, -CH2CH2SCH3, CH2CO2H, CH2C(O)NH2, CH2CH2COOH, CH2CH2C(O)NH2, CH2CH2CH2CH2NH2, -CH2CH2CH2NHC(NH)NH2, CH2-imidazole-4-yl, CH2OH, CH(OH)CH3, CH2((4'-OH)Ph), CH2SH, or lower cycloalkyl, R 9b It is hydrogen, R 10 C may be substituted with hydrogen, alkoxy, di(lower alkyl)-amino, or halogen. 1~6 Alkyl, C 1~6 Haloalkyl, C 3~7 These include cycloalkyl, heterocycloalkyl, aminoacyl, aryl such as phenyl, heteroaryl such as pyridinyl, substituted aryl, or substituted heteroaryl. R 11 C may be substituted. 1~6 alkyl, possibly substituted cycloalkyl, possibly substituted C 2~6 Alkinyl substitution, KidC 2~6 Alkenyl or substituted acyl, but not limited to C(O)(C1~6 Contains alkyl, Y, R 3 , R 12 and R 22 This is by means of (as defined herein).

[0073] In one embodiment, formula IV: [ka] (In the formula, variables Y and R 3 , R 7 , R 8 , R 9a , R 9b , R 10 and R 22 This specification discloses compounds of the type described herein.

[0074] In one embodiment, formula V: [ka] (In the formula, variables Y and R 3 , R 7 , R 8 , R 9a , R 9b , R 10 and R 22 This specification discloses compounds of the type described herein.

[0075] In alternative embodiments, compounds, methods, and compositions for the treatment of a host infected with or exposed to hepatitis C are provided.

[0076] In one embodiment, formula VI: [ka] (In the formula, R 41 This includes halogens (especially F or Cl), OR 3 (Containing OH), N3, NH2, or CN, and also variables Y, R 3 , R 4 and R 12 This specification discloses compounds of the type described herein.

[0077] In one embodiment, formula VII: [ka] (In the formula, variables Y and R 3 , R 4 , R 12 and R 41 This specification discloses compounds of the type described herein.

[0078] β-D-2'-deoxy-2'-α-fluoro-2'-β-C-substituted-N 6 Metabolism of -substituted-2,6-diaminopurine nucleotides β-D-2'-deoxy-2'-α-fluoro-2'-β-methyl-N as a phosphoramidate 6 The metabolism of methyl-2,6-diaminopurine nucleoside involves the production of 5'-monophosphate and N-2'-deoxy-2'-α-fluoro-2'-β-methyl-guanine nucleoside as 5'-monophosphate. 6 This involves the subsequent assimilation of a methyl-2,6-diaminopurine base. The monophosphate is further assimilated into the active species, namely the 5'-triphosphate. Here, β-D-2'-deoxy-2'-α-fluoro-2'-β-methyl-guanine triphosphate is IC10 against HCV genotype 1b NS5B polymerase. 50 It has a concentration of 0.15 μM. β-D-2'-deoxy-2'-α-fluoro-2'-β-methyl-N 6 The metabolic pathway for -methyl-2,6-diaminopurine nucleoside phosphoramidate is explained in mechanism 1 below. [ka]

[0079] β-D-2'-deoxy-2'-α-fluoro-2'-β-methyl-N 6 The metabolism of -dimethyl-2,6-diaminopurine nucleotide is β-D-2'-deoxy-2'-α-fluoro-2'-β-methyl-N 6This involves both the formation of -dimethyl-2,6-diaminopurine nucleoside triphosphate and the production of the corresponding guanine nucleoside triphosphate. The metabolic pathways are shown in mechanisms 2 and 3 below. [ka] JPEG0007893504000040.jpg122169

[0080] Stabilized Phosphate Prodrug A stabilized phosphate prodrug is a portion that can deliver a monophosphate, diphosphate, or triphosphate in vivo. For example, McGuigan disclosed phosphoramidates in U.S. Patents 8,933,053, 8,759,318, 8,658,616, 8,263,575, 8,119,779, 7,951,787 and 7,115,590. Alios cites By means of this specification, thiophosphoramidates were disclosed in U.S. Patent Nos. 8,895,723 and 8,871,737. Alios also cited by means of Therefore, cyclic nucleotides are disclosed in U.S. Patent No. 8,772,474, which is part of this specification. Idenix discloses cyclic phosphoramidates and phosphoramidate / SATE derivatives in International Publication No. 2013 / 177219, which is part of this specification by reference. Idenix also discloses substituted carbonyloxymethylphosphoramidate compounds in International Publication No. 2013 / 039920, which is part of this specification by reference. Hostetler has disclosed lipid phosphate prodrugs. For example, See U.S. Patent No. 7,517,858. Hostetler also offers phosphonate pro Lipid complexes of drugs have been disclosed. See, for example, U.S. Patents 8,889,658, 8,846,643, 8,710,030, 8,309,565, 8,008,308, and 7,790,703. Emory University disclosed nucleotide sphingoids and lipid derivatives in International Publication 2014 / 124430. RFS Pharma disclosed purine nucleoside monophosphate prodrugs in International Publication 2010 / 091386. Cocrystal Pharma Inc. also disclosed, By doing so, a purine nucleoside monophosphate prodrug was disclosed in U.S. Patent No. 9,173,893, which forms part of this specification. HepDirect® technology was disclosed in the paper "Design, Synthesis, and Characterization of a Series of Cytochrome P(450) 3A-Activated Prodrugs (HepDirect Prodrugs) Useful for Targeting Phosph(on)ate-Based Drugs to the Liver," (J. Am. Chem. SoC.126, 5154-5163 (2004)). Further phosphate prodrugs include, but are not limited to, phosphate esters, 3',5'-cyclic phosphates including CycloSAL, SATE derivative (S-acyl-2thioester) prodrugs, and DTE (dithiodiethyl) prodrugs. For a review of literature disclosing non-limiting examples, see A. Ray and K. Hostetler, "Application of kinase bypass strategies to nucleoside antivirals," Antiviral Research (2011) 277-291, M. Sofia, "Nucleotide prodrugs for HCV therapy," Antiviral Chemistry and Chemotherapy 2011; 22-23-49, and S. Peyrottes et al., "SATE Pronucleotide Approaches: An Overview," Mini Reviews in Medicinal Chemistry 2004, 4, 395. In one embodiment, a 5'-prodrug described in any of these patent applications or literature is used as the R of the presented compound. 4 It can be used for rank.

[0081] In an alternative embodiment, stabilized phosphate prodrugs include, but are not limited to, those described in U.S. Patents 9,173,893 and 8,609,627, which are part of this specification by reference (including methods for preparation). For example, 5'-prodrugs of formulas I to V may be represented by the following groups. [ka]

[0082] In alternative embodiments, the 3',5'-prodrugs of formulas I to V may be represented by the following groups: [ka] (In the formula, When chirality is present at the center of phosphorus, it is R, either entirely or partially. p Or S p It may be either one of those, or any mixture thereof. Z is either O or S, R 33 is OR 34 , [ka] and derived fatty alcohols (e.g., but not limited to, [ka] Selected from, In the formula, R 34 , R 35 and R 36 It is defined as follows: R 31 and R 32 When administered in vivo, it can provide a nucleoside monophosphate or thiomonophosphate, which may or may not be partially or completely resistant to 6-NH2 deamination in the biological system. Representative R 31 and R 32 The following are selected independently: (a)OR 34 , here, R 34 If is selected from H, Li, Na, K, phenyl and pyridinyl, then phenyl and pyridinyl are (CH2) 0~6 CO2R 37 and (CH2) 0~6 CON(R 37 ) Substituted with 1 to 3 substituents independently selected from the group consisting of 2, R 37 H and C are independent of each other. 1~20 Alkyl, fatty alcohol (oleyl alcohol, octacosanol, triacontanol, linoleyl alcohol, etc.) carbon chains, or lower alkyl, alkoxy, di(lower alkyl)-amino, fluoro, C 3~10Cycloalkyl, cycloalkylalkyl, cycloheteroalkyl, aryl such as phenyl, heteroaryl such as pyridinyl, substituted aryl, or substituted heteroaryl C 1~20 It is alkyl, Here, the substituent is C 1~5 Alkyl, or lower alkyl, alkoxy, di(lower alkyl)-amino, fluoro, C 3~10 Cycloalkyl or cycloalkyl-substituted C 1~5 It is alkyl. (b) [ka] (c) Esters of D-amino acids or L-amino acids [ka] In the formula, R 36 This is limited to side chains that occur in natural L-amino acids. R 35 H, C 1~20 Alkyl, fatty alcohol (oleyl alcohol, octacosanol, triacontanol, linoleyl alcohol, etc.) carbon chains, or lower alkyl, alkoxy, di(lower alkyl)-amino, fluoro, C 3~10 Cycloalkyl, cycloalkylalkyl, cycloheteroalkyl, aryl such as phenyl, heteroaryl such as pyridinyl, substituted aryl, or substituted heteroaryl C 1~20 It is alkyl, Here, the substituent is C 1~5 Alkyl, or lower alkyl, alkoxy, di(lower alkyl)-amino, fluoro, C 3~10 Cycloalkyl or cycloalkyl-substituted C 1~5 It is alkyl. (d)R 31 and R 32 They may together form a ring, [ka] In the formula, R 38H, C 1~20 Alkyl, C 1~20 Carbon chains derived from alkenyls, fatty alcohols (oleyl alcohol, octacosanol, triacontanol, linoleyl alcohol, etc.), or lower alkyl, alkoxy, di(lower alkyl)-amino, fluoro, C 3~10 Cycloalkyl, cycloalkylalkyl, cycloheteroalkyl, aryl such as phenyl, heteroaryl such as pyridinyl, substituted aryl, or C substituted with substituted heteroaryl 1~20 It is alkyl, Here, the substituent is C 1~5 Alkyl, or lower alkyl, alkoxy, di(lower alkyl)-amino, fluoro, C 3~10 Cycloalkyl or cycloalkyl-substituted C 1~5 It is alkyl. (e)R 31 and R 32 They may together form a ring selected from the following: [ka] During the ceremony, R 39 is O or NH, R 40 H, C 1~20 Alkyl, C 1~20 Alkenyl, carbon chain derived from fatty acids (oleic acid, linoleic acid, etc.), and lower alkyl, alkoxy, di(lower alkyl)-amino, fluoro, C 3~10 Cycloalkyl, cycloalkylalkyl, cycloheteroalkyl, aryl such as phenyl, heteroaryl such as pyridinyl, substituted aryl, or substituted heteroaryl C 1~20 It is alkyl, Here, the substituent is C 1~5 Alkyl, or lower alkyl, alkoxy, di(lower alkyl)-amino, fluoro, C 3~10 Cycloalkyl or cycloalkyl-substituted C 1~5 It is alkyl.

[0083] The above compounds can be prepared, for example, by preparing 5'-OH analogs and then converting them to monophosphate analogs.

[0084] Embodiment In a particular embodiment, (i) In equation Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 is hydrogen, R 3 is hydrogen, R 4 It is a stabilized phosphate prodrug. (ii) In equation Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 is hydrogen, R 3 is hydrogen, R 4 It is a stabilized thiophosphate prodrug. (iii) In equation Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 is hydrogen, R 3 is hydrogen, R 4 It is a phosphoamidate. (iv) In equation Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 is hydrogen, R 3 is hydrogen, R 4 It is a thiophosphorumidate. (v) In equation Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 is hydrogen, R 3 is hydrogen, R 4 It is a monophosphate. (vi) In equation Ia, Y is NR 1 R 2 And R 1 is methyl, and R2 is hydrogen, R 3 is hydrogen, R 4 It is a diphosphate. (vii) In equation Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 is hydrogen, R 3 is hydrogen, R 4 It is a triphophen. (viii) In equation Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 is methyl, and R 3 is hydrogen, R 4 It is a stabilized phosphate prodrug. (ix) In equation Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 is methyl, and R 3 is hydrogen, R 4 It is a stabilized thiophosphate prodrug. (x) In equation Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 is methyl, and R 3 is hydrogen, R 4 It is a phosphoamidate. In equation (xi) Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 is methyl, and R 3 is hydrogen, R 4 It is a thiophosphorumidate. (xii) In equation Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 is methyl, and R 3 is hydrogen, R 4 It is a monophosphate. (xiii) In equation Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 is methyl, and R 3 is hydrogen, R 4 It is a diphosphate. (xiv) In equation Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 is methyl, and R 3 is hydrogen, R 4 It is a triphophen. In equation (xv) Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 It is cyclopropyl, and R 3 is hydrogen, R 4 It is a stabilized phosphate prodrug. In equation (xvi) Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 It is cyclopropyl, and R 3 is hydrogen, R 4 It is a stabilized thiophosphate prodrug. In equation (xvii) Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 It is cyclopropyl, and R 3 is hydrogen, R 4 It is a phosphoamidate. In equation (xviii) Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 It is cyclopropyl, and R 3 is hydrogen, R 4 It is a thiophosphorumidate. In equation (xix) Ia, Y is NR 1 R2 And R 1 is methyl, and R 2 It is cyclopropyl, and R 3 is hydrogen, R 4 It is a monophosphate. In equation (xx)Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 It is cyclopropyl, and R 3 is methyl, and R 4 It is a diphosphate. In equation (xxi)Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 It is cyclopropyl, and R 3 is hydrogen, R 4 It is a triphophen. In equation (xxii)Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 It is propyl, and R 3 is hydrogen, R 4 It is a stabilized phosphate prodrug. In equation (xxiii)Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 It is propyl, and R 3 is hydrogen, R 4 It is a stabilized thiophosphate prodrug. In formula (xxiv) Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 It is propyl, and R 3 is hydrogen, R 4 It is a phosphoamidate. In equation (xxv)Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 It is propyl, and R3 is hydrogen, R 4 It is a thiophosphorumidate. In equation (xxvi)Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 It is propyl, and R 3 is hydrogen, R 4 It is a monophosphate. In equation (xxvii)Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 It is propyl, and R 3 is hydrogen, R 4 It is a diphosphate. In equation (xxviii)Ia, Y is NR 1 R 2 And Y is NR 1 R 2 And R 1 is methyl, and R 2 It is propyl, and R 3 is hydrogen, R 4 It is a triphophen. In equation (xxix) Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 is ethyl, and R 3 is hydrogen, R 4 It is a stabilized phosphate prodrug. In equation (xxx)Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 is ethyl, and R 3 is hydrogen, R 4 It is a stabilized thiophosphate prodrug. In equation (xxxi)Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 is ethyl, and R 3 is hydrogen, R4 It is a phosphoamidate. In equation (xxxii)Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 is ethyl, and R 3 is hydrogen, R 4 It is a thiophosphorumidate. In equation (xxxiii)Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 is ethyl, and R 3 is hydrogen, R 4 It is a monophosphate. In equation (xxxiv) Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 is ethyl, and R 3 is hydrogen, R 4 It is a diphosphate. In equation (xxxv)Ia, Y is NR 1 R 2 And R 1 is methyl, and R 2 is ethyl, and R 3 is hydrogen, R 4 It is a triphophen. In equation (xxxvi) Ib, Y is NR 1 R 2 And R 1 is methyl, and R 2 is methyl, and R 3 is hydrogen, R 4 It is a stabilized phosphate prodrug. In formula (xxxvii) Ib, Y is NR 1 R 2 And R 1 is methyl, and R 2 is methyl, and R 3 is hydrogen, R 4 It is a stabilized thiophosphate prodrug. In equation (xxxviii) Ib, Y is NR 1 R 2 And R 1 is methyl, and R 2 is methyl, and R 3 is hydrogen, R 4 It is a phosphoamidate. In equation (xxxix) Ib, Y is NR 1 R 2 And R 1 is methyl, and R 2 is methyl, and R 3 is hydrogen, R 4 It is a thiophosphorumidate. In equation (xl) Ib, Y is NR 1 R 2 And R 1 is methyl, and R 2 is methyl, and R 3 is hydrogen, R 4 It is a monophosphate. In equation (xli) Ib, Y is NR 1 R 2 And R 1 is methyl, and R 2 is methyl, and R 3 is hydrogen, R 4 It is a diphosphate. In equation (xlii) Ib, Y is NR 1 R 2 And R 1 is methyl, and R 2 is Methi It is R 3 is hydrogen, R 4 It is a triphophen. In equation (xliii) Ib, Y is NR 1 R 2 And R 1 is methyl, and R 2 is hydrogen, R 3 is hydrogen, R 4 It is a stabilized phosphate prodrug. In equation Ib (xliv), Y is NR 1 R 2 And R1 is methyl, and R 2 is hydrogen, R 3 is hydrogen, R 4 It is a stabilized thiophosphate prodrug. In equation (xlv) Ib, Y is NR 1 R 2 And R 1 is methyl, and R 2 is hydrogen, R 3 is hydrogen, R 4 It is a phosphoamidate. In equation (xlvi) Ib, Y is NR 1 R 2 And R 1 is methyl, and R 2 is hydrogen, R 3 is hydrogen, R 4 It is a thiophosphorumidate. In equation (xlvii) Ib, Y is NR 1 R 2 And R 1 is methyl, and R 2 is hydrogen, R 3 is hydrogen, R 4 It is a monophosphate. In equation (xlviii) Ib, Y is NR 1 R 2 And R 1 is methyl, and R 2 is hydrogen, R 3 is hydrogen, R 4 It is a diphosphate. In equation Ib (xlix), Y is NR 1 R 2 And R 1 is methyl, and R 2 is hydrogen, R 3 is hydrogen, R 4 It is a triphophen. (l) In equation Ib, Y is NR 1 R 2 And R 1 is methyl, and R 2 It is cyclopropyl, and R 3 is hydrogen, R4 is a stabilized phosphate prodrug. (li) In formula Ib, Y is NR 1 R 2 where R 1 is methyl, R 2 is cyclopropyl, R 3 is hydrogen, R 4 is a stabilized thiophosphate prodrug. (lii) In formula Ib, Y is NR 1 R 2 where R 1 is methyl, R 2 is cyclopropyl, R 3 is hydrogen, R 4 is phosphoramidate. (liii) In formula Ib, Y is NR 1 R 2 where R 1 is methyl, R 2 is cyclopropyl, R 3 is hydrogen, R 4 is thiophosphoramidate. (liv) In formula Ib, Y is NR 1 R 2 where R 1 is methyl, R 2 is cyclopropyl, R 3 is hydrogen, R 4 is monophosphate. (lv) In formula Ib, Y is NR 1 R 2 where R 1 is methyl, R 2 is cyclopropyl, R 3 is methyl, R 4 is diphosphate. (lvi) In formula Ia, Y is NR 1 R 2 where R 1 is methyl, R 2 is cyclopropyl, R 3 is hydrogen, R 4 is triphosphate.

[0085] In an alternative embodiment of any of the above, the compound has an R 22 substituent. In any of these particular embodiments, R 22 is F, an amide or a carbamate. In other particular aspects of the above embodiments, R 22 is chloro, bromo, cyano, azido, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, tert-butyl and n-pentyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, 3-methylbutyl, 1-methylbutyl, 1-ethylpropyl, vinyl, allyl, 1-butynyl, 2-butynyl, ethynyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, -(CH2)-cyclopropyl, -(CH2)-cyclobutyl, -(CH2)-cyclopentyl, -(CH2)-cyclohexyl, aziridine, oxirane, thiirane, azetidine, oxetane, thietane, pyrrolidine, tetrahydrofuran, thiolane, pyrazolidine, piperidine, oxane, thiane, -(CH2)-aziridine, -(CH2)-oxirane, -(CH2)-thiirane, -(CH2)-azetidine, -(CH2)-oxetane, -(CH2)-thietane, -(CH2)-pyrrolidine, -(CH2)-tetrahydrofuran, -(CH2)-thiolane, -(CH2)-pyrazolidine, -(CH2)-piperidine, -(CH2)-oxane, -(CH2)-thiane, phenyl, pyridyl, -ONHC(=O)OCH3, -ONHC(=O)OCH2CH3, -NHOH, NHOCH3, -OCH3, OC2H5, -OPh, OCH2Ph, -SCH3, -SC2H5, -SPh, SCH2Ph, -NH(CH2)2NH2, -NH(CH2)2N(CH3)2, -NHNH2, -NHNHCH3, -N=NH, -N=NCH3, -N=NCH2CH3, -NHC(O)NHNH2, -NHC(S)NHNH2, -C(O )NHNH2, -NHSO2CH3, -NHSO2CH2CH3, -SO2NHCH3, -SO2N(CH3)2, -C(O)NH2, -C(O)NHCH3, -C(O)N(CH3)2, -CO2CH3, -CO2CH2CH3, -CO2Ph, CO2CH2Ph, -SO2CH3, -SO2CH2CH3, -SO2Ph, -SO2CH2Ph, [ka] , -P(O)H(OH), -P(O)H(OCH3), -P(O)(OH)(OH), -P(O)(OH)(OCH3), -P(O)(OCH3)(O CH3), -P(O)(OH)(NH2), -P(O)(OH)(NHCH3), -P(O)(OH)N(CH3)2, -NHC(O)CH3, -NHC (O)CH2CH3, -NHC(O)CH(CH3)2, -NHC(O)OCH3, -NHC(O)OCH2CH3, -NHC(O)OCH(CH3)2, -NHC(O)OCH2CH2CH3, -NHC(O)OCH2CH2CH2CH3 and -NHC(O)OCH2CH2CH2CH2CH3.

[0086] In alternative embodiments of compounds (i) to (lvi), L-nucleosides are used in formulas I to VII.

[0087] In an alternative embodiment, R in formula I 12 The variable is CH2F.

[0088] In an alternative embodiment, R in formula I 12 The variable is CHF2.

[0089] In an alternative embodiment, R in formula I 12 The variable is CF3.

[0090] In one embodiment, a compound of formula Ia is provided. Non-limiting examples of compounds of formula Ia include the following: [ka] JPEG0007893504000051.jpg211169JPEG0007893504000052.jpg210169JPEG0007893504000053.jpg83169

[0091] In one embodiment, a thiophosphoramidate of formula Ia is provided. Examples of thiophosphoramidates of formula Ia, but not limited to these, include: [ka] JPEG0007893504000055.jpg207169JPEG0007893504000056.jpg211169JPEG0007893504000057.jpg125169

[0092] In one embodiment, a stabilized phosphate prodrug of formula Ia is provided. Non-limiting examples of a stabilized phosphate prodrug of formula Ia include the following: [ka] JPEG0007893504000059.jpg84169

[0093] In another embodiment, compounds of formula Ia are provided. Non-limiting examples of compounds of formula Ia include the following: [ka] JPEG0007893504000061.jpg216169JPEG0007893504000062.jpg216169JPEG0007893504000063.jpg214169

[0094] In one embodiment, a thiophosphoramidate of formula Ia is provided. Examples of thiophosphoramidates of formula Ia, but not limited to these, include: [ka] JPEG0007893504000065.jpg204169JPEG0007893504000066.jpg164169JPEG0007893504000067.jpg86169

[0095] In one embodiment, a stabilized phosphate prodrug of formula Ia is provided. Non-limiting examples of a stabilized phosphate prodrug of formula Ia include the following: [ka] JPEG0007893504000069.jpg153169

[0096] In one embodiment, a compound of formula II is provided. Non-limiting examples of compounds of formula II include the following: [ka] JPEG0007893504000071.jpg187169JPEG0007893504000072.jpg185169JPEG00078935040 00073.jpg189169JPEG0007893504000074.jpg213169JPEG0007893504000075.jpg167169

[0097] In one embodiment, a compound of formula I is provided. Non-limiting examples of compounds of formula I include the following: [ka] JPEG0007893504000077.jpg176169JPEG0007893504000078.jpg91169

[0098] In one embodiment, a compound of formula II is provided. Non-limiting examples of compounds of formula II include the following: [ka]

[0099] In one embodiment, R 4 teeth, [ka] That is the case.

[0100] In one embodiment, a compound of formula II is provided. Non-limiting examples of compounds of formula II include the following: [ka] JPEG0007893504000082.jpg219169JPEG0007893504000083.jpg189169JPEG0007893504000084.jpg196169JPEG0007893504000085.jpg119169

[0101] In some embodiments, R 3 H is R 4 teeth, [ka] That is the case.

[0102] In some embodiments, R 3 H is R 4 teeth, [ka] That is the case.

[0103] In some embodiments, R 3 H is R 4 teeth, [ka] That is the case.

[0104] In one embodiment, a compound of formula II is provided. Non-limiting examples of compounds of formula II include the following: [ka] JPEG0007893504000090.jpg179169JPEG0007893504000091.jpg179169JPEG000 7893504000092.jpg216169JPEG0007893504000093.jpg181169JPEG00078935040 00094.jpg217169JPEG0007893504000095.jpg218169JPEG0007893504000096.j pg216169JPEG0007893504000097.jpg219169JPEG0007893504000098.jpg213169

[0105] In some embodiments, R 3 H is R 4 teeth, [ka] That is the case.

[0106] In some embodiments, R 3 H is R 4 teeth, [ka] That is the case.

[0107] In some embodiments, R 3 H is R 4 teeth, [ka] That is the case.

[0108] In some embodiments, R 1 It is CH3, and R 2 H is R 3 H is R 4 teeth, [ka] That is the case.

[0109] In some embodiments, R 1 It is CH3, and R 2 H is R 3 H is R 4 teeth, [ka] That is the case.

[0110] In some embodiments, R 1 It is CH3, and R 2 H is R 3 H is R 4 teeth, [ka] That is the case.

[0111] In some embodiments, R 1 It is CH3, and R 2 It is CH3, and R 3 H is R 4 teeth, [ka] That is the case.

[0112] In some embodiments, R 1 It is CH3, and R 2 It is CH3, and R 3 H is R 4 teeth, [ka] That is the case.

[0113] In some embodiments, R 1 It is CH3, and R 2 It is CH3, and R 3 H is R4 teeth, [ka] That is the case.

[0114] In some embodiments, R 1 It is cyclopropyl, and R 2 It is CH3, and R 3 H is R 4 teeth, [ka] That is the case.

[0115] In some embodiments, R 1 It is cyclopropyl, and R 2 It is CH3, and R 3 H is R 4 teeth, [ka] That is the case.

[0116] In some embodiments, R 1 It is cyclopropyl, and R 2 It is CH3, and R 3 H is R 4 teeth, [ka] That is the case.

[0117] II. Definition The following terms are used to describe the present invention. Any term specifically defined herein is... If not, the term shall be given a meaning that is recognizable to a person skilled in the art when applying the term to the context in which it is used to describe the present invention.

[0118] In this context, the term "alkyl" means a linear or branched, fully saturated, optionally substituted (e.g., by a halogen containing fluorine) hydrocarbon radical or alkyl group. For example, an alkyl group may have one, two, three, four, five, six, seven, or eight carbon atoms (i.e., C1-C8 alkyl), one, two, three, four, five, or six carbon atoms (i.e., C1-C6 alkyl), or one to four carbon atoms (i.e., C1-C4 alkyl). Suitable examples of alkyl groups, but not limited to, include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, tert-pentyl, neopentyl, hexyl, 2-methylpentyl, 3-methylpentyl, 2,2-dimethylbutyl, and 2,3-dimethylbutyl.

[0119] The term "alkenyl" refers to a non-aromatic hydrocarbon group that contains at least one double bond between adjacent carbon atoms and, unless otherwise specified herein, an alkyl group-like structure. For example, an alkenyl group may have 2 to 8 carbon atoms (i.e., C2-C8 alkenyls) or 2 to 4 carbon atoms (i.e., C2-C4 alkenyls). Suitable examples of alkenyl groups, but not limited to, include ethenyl or vinyl (-CH=CH2), allyl (-CH2CH=CH2), 1-butenyl (-C=CH-CH2CH3), and 2-butenyl (-CH2CH=CHCH2). As described herein, the alkenyl group may be substituted.

[0120] The term "alkynyl" refers to a non-aromatic hydrocarbon group that contains at least one triple bond between adjacent carbon atoms and, unless otherwise specified herein, an alkyl group-like structure. For example, an alkynyl group may have 2 to 8 carbon atoms (i.e., C2-C8 alkynes) or 2 to 4 carbon atoms (i.e., C2-C4 alkynyls). Examples of alkynyl groups, but not limited to, include acetylenyl, ethynyl, and propargyl. As described herein, the alkynyl group may be substituted.

[0121] The term "acyl" refers to a moiety -C(O)R in which the carbonyl portion is bonded to R, for example, a -C(O) alkyl group. R may be selected from alkoxy, alkyl, cycloalkyl, lower alkyl (i.e., C1-C4), alkoxyalkyl including methoxymethyl, aralkyl including benzyl, aryloxyalkyl including phenoxymethyl, halogen, C1-C4 alkyl, or aryl including phenyl which may be substituted with a C1-C4 alkoxy. In one embodiment, the term "acyl" refers to a monophosphate, diphosphate, or triphophate.

[0122] The term "lower acyl" refers to an acyl group in which the carbonyl portion is a lower alkyl group (i.e., C1-C4).

[0123] The term "alkoxy" refers to the group -OR', where -OR' is -O-alkyl, -O-alkenyl, -O-alkynyl, -O-(C0~C2)(cycloalkyl), -O-(C0~C2)(heterocyclo), -O-(C0~C2)(aryl), or -O-(C0~C2)(heteroaryl), and each of these may be substituted.

[0124] The term "amino" refers to the group -NH2.

[0125] The terms "amino acid" or "amino acid residue" refer to D- or L-amino acids of natural or unnatural origin. This refers to amino acids. Representative amino acids include, but are not limited to, alanine, β-alanine, arginine, asparagine, aspartic acid, cysteine, cystine, glutamic acid, glutamine, glycine, phenylalanine, histidine, isoleucine, lysine, leucine, methionine, proline, serine, threonine, valine, tryptophan, or tyrosine.

[0126] The term "Azido" refers to base-N3.

[0127] The terms “aryl” or “aromatic” in this context refer to a monovalent aromatic radical having a single substituted (unless otherwise specified herein) or unsubstituted ring (e.g., phenyl or benzyl), or a fused ring (e.g., naphthyl, anthracenyl, phenantrenyl, etc.), which may be bonded to the compounds according to the present invention at any available stable position on the ring(s), or otherwise as shown in the presented chemical structures. The aryl group may be substituted as described herein.

[0128] "Cycloalkyl," "carbocyclic," or "carbocyclyl" refers to saturated ( That is, it refers to a ring having 3 to 7 carbon atoms as a monocyclic (cycloalkyl) or partially unsaturated (e.g., cycloalkenyl, cycloalkadienyl, etc.) ring. Monocyclic carbocyclic rings have 3 to 7 ring atoms, more typically 5 or 6 ring atoms. Non-limiting examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopenta-1-enyl, 1-cyclopenta-2-enyl, 1-cyclopenta-3-enyl, cyclohexyl, 1-cyclohexa-1-enyl, 1-cyclohexa-2-enyl, and 1-cyclohexa-3-enyl.

[0129] The term "cyano" refers to the group -CN.

[0130] The terms "halogen" or "halo" refer to chloro, bromo, fluoro, or iodine.

[0131] A heteroaryl ring system is a saturated or unsaturated ring having one or more nitrogen, oxygen, or sulfur atoms in a (mono) ring, but is not limited to this, and particularly includes condensed ring systems such as imidazole, furyl, pyrrole, furanyl, thien, thiazole, pyridine, pyrimidine, purine, pyrazine, triazole, oxazole, or indole, quinoline, etc., which may be optionally substituted as described above. Examples of heteroaryl groups include nitrogen-containing heteroaryl groups, such as pyrrole, pyridine, pyridone, pyridazine, pyrimidine, pyrazine, pyrazole, imidazole, triazole, triazine, tetrazole, indole, isoindole, indidine, purine, indazole, quinoline, isoquinoline, quinoridine, phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline, pteridine, imidazopyridine, imidazotriazine, pyrazinopyridazine, acridine, phenanthridine, carbazole, carbazoline, perimidine, phenanthroline, phenacene, oxadiazole, benzimidazole, pyrrolopyridine, and pyrrolopyrimidine. and pyridopyrimidines; sulfur-containing aromatic heterocycles, e.g., thiophene and benzothiophene; oxygen-containing aromatic heterocycles, e.g., furan, pyran, cyclopentapyran, benzofuran and isobenzofuran; as well as aromatic heterocycles containing two or more heteroatoms selected from nitrogen, sulfur and oxygen, e.g., thiazole, thiadizole, isothiazole, benzoxazole, benzothiazole, benzothiadiazole, phenothiazine, isoxazole, furazan, phenoxazine, pyrazoloxazole, imidazothiazole, thienofuran, furopyrrole, pyridoxazine, phlopyrrole Examples include dins, phlopyrimidines, thienopyrimidines, and oxazoles, all of which may be substituted as desired.

[0132] The terms "heterocyclic" or "heterocyclic" refer to a cyclic group containing at least one heteroatom, i.e., O, N, or S, which may be aromatic (heteroaryl) or non-aromatic. Examples of non-aromatic heterocyclic groups used in the present invention include, for example, pyrrolidinyl, piperidinyl, piperazinyl, N-methylpiperazinyl, imidazolinyl, pyrazolidinyl, imidazolidinyl, morpholinyl, tetrahydropyranyl, azetidinyl, oxetanyl, oxathiolanil, pyridone, 2-pyrrolidone, ethyleneurea, 1,3-dioxolane, 1,3-dioxane, 1,4-dioxane, phthalimide, and succinimide, all of which may be optionally substituted.

[0133] The term "hydroxyl" refers to the -OH group.

[0134] The term "nitro" refers to the group -NO2.

[0135] The terms "pharmaceutically acceptable salt" or "prodrug" refer to a β-D-2'-D-2'-α-fluoro-2'-β-C-substituted-2-modified-N compound that provides the desired active compound upon administration to a patient. 6- Used throughout the specification to describe any pharmaceutically acceptable form of substituted purine nucleotides (esters, phosphoramides, thiophosphoramides, phosphate esters, ester salts, or related groups). Examples of pharmaceutically acceptable salts are organic acid addition salts formed by acids that form physiologically acceptable anions, such as tosylates, methanesulfonates, acetates, citrates, malons, tartrates, succinates, benzoates, ascorbicates, α-ketoglutarates, and α-glycerophosphates. Suitable inorganic salts, including salts of sulfates, nitrates, bicarbonates, and carbonates, can also be formed. Pharmaceutically acceptable salts can be obtained by reacting a sufficiently basic compound, such as an amine, with a suitable acid that forms a physiologically acceptable anion, using standard methods well known in the art. Salts of alkali metals (e.g., sodium, potassium, or lithium) or alkaline earth metals (e.g., calcium) of carboxylic acids can also be made.

[0136] A "pharmaceutically acceptable prodrug" refers to a compound that is metabolized in a host, for example, by hydrolysis or oxidation, to form the compound of the present invention. Typical examples of prodrugs include compounds having a biologically unstable protecting group in the functional portion of the active compound. Examples of prodrugs include compounds that can be oxidized, reduced, aminated, deaminated, hydroxylated, dehydroxylated, hydrolyzed, dehydrolyzed, alkylated, dealkylated, acylated, deacylated, phosphorylated, dephosphorazonated, thiophosphorazonated, dethiophosphorazonated, phosphoramidated, or dephosphorazonated to produce the active compound. The compound of the present invention has antiviral activity against HCV, or is metabolized to a compound exhibiting such activity. β-D-2'-D-2'-α-fluoro-2'-β-C-substituted-2-modified-N 6- The replacement purine nucleosides can also be administered as 5'-phosphoether lipids, bisphosphoramidates, 3',5'-cyclic phosphoramidates, 3',5'-cyclic thiophosphoramidates, DTE conjugates, mixed phosphoramidate-SATE derivatives, or "SATE" derivatives.

[0137] The term "phosphoric acid" refers to the group -P(O)(OH)2.

[0138] In one embodiment, the terms purine or pyrimidine base include, but are not limited to, adenine, N 6 -alkylpurine, N 6 -acylpurine (where acyl is -C(O)alkyl, -C(O)(aryl)C0-C4 alkyl, or -C(O)(C0-C4 alkyl)aryl), N 6 -benzylpurine, N 6 -halopurine, N 6 -vinylpurine, N 6 -acetylenepurine, N 6 -acylpurine, N 6 -hydroxyalkylpurine, N 6 -thioalkylpurine, N 2 -alkylpurine, N 2 -alkyl-6-thiopurine, thymine, si tosine, 5-fluorocytosine, 5-methylcytosine, 6-azapyrimidines including 6-azacytosine, 2- and / or 4-mercaptopyrimidines, uracil, 5-halouracils including 5-fluorouracil, C 5 -alkylpyrimidine, C 5 -benzylpyrimidine, C 5 -halopyrimidine, C 5 -vinylpyrimidine, C 5 -acetylenepyrimidine, C 5 -acylpyrimidine, C 5 -hydroxyalkylpurine, C 5 -amidopyrimidine, C 5 -cyanopyrimidine, C 5 -nitropyrimidine, C 5 -aminopyrimidine, N 2-Alkylpurine, N 2 These include alkyl-6-thiopurines, 5-azacitidinyl, 5-azaurasilyl, triazolopyridinyl, imidazolopyridinyl, pyrrolopyrimidinyl, and pyrazolopyrimidinyl. Examples of purine bases, but not limited to, include guanine, adenine, hypoxanthine, 2,6-diaminopurine, and 6-chloropurine. Functional oxygen and nitrogen groups on the base can be protected as needed or desired. Suitable protecting groups, well known to those skilled in the art, include benzyl, trimethylsilyl, dimethylhexylsilyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, trityl, alkyl groups, and acyl groups such as acetyl and propionyl, methanesulfonyl, and p-toluenesulfonyl. Alternatively, the purine or pyrimidine base may be substituted to form a viable prodrug that can be cleaved in vivo. Examples of suitable substituents include acyl moieties.

[0139] The terms “substituted” or “may be substituted” indicate that the part may have at least one additional substituent, including, but not limited to, halogens (F, Cl, Br, I), OH, phenyl, benzyl, N3, CN, acyl, alkyl, alkenyl, alkynyl, alkoxy, CHF2, CH2F, and CF3, haloalkyl, etc. In one embodiment, the terms “substituted” or “may be substituted” indicate that the part may have at least one additional substituent, including, but not limited to, azide, cyano, halogen (fluoro, chloro, bromo, or iodine), alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, heteroaryl, haloalkyl, hydroxyl, alkoxy, amino, -NH(C1~C6 unsubstituted alkyl), -NH(C1~C6 substituted alkyl), -NH-(C0~C2 alkyl)(C3~C8 cycloalkyl), -NH-(C0~C2 alkyl)(C3~C8 heterocyclic), This indicates that the molecule may have at least one additional substituent, including NH-(C0-C2 alkyl)(aryl), -N(C1-C6 unsubstituted alkyl)2, -N(C1-C6 unsubstituted alkyl)(C1-C6 substituted alkyl), -N(C1-C6 substituted alkyl)2, -NH-(C0-C2 alkyl)(C3-C8 cycloalkyl), -NH-(C0-C2 alkyl)(C3-C8 heterocyclic), -NH-(C0-C2 alkyl)(aryl), acyl, nitro, sulfonic acid, sulfate, phosphonic acid, phosphate, phosphonate, or thiol.

[0140] formula R 14 S(O)2OR 15 The term "sulfonic acid ester" as expressed by R 14 Including, here, R 14 R is alkyl, haloalkyl, aralkyl, or aryl. 15 These are alkyl, aryl, or aralkyl compounds.

[0141] The term "sulfonic acid" refers to the group -SO2OH.

[0142] The term "thiol" refers to the group -SH.

[0143] As used herein, the term “nitrogen protecting group” refers to a moiety that is covalently bonded to nitrogen and is removable, typically substituted with hydrogen where appropriate. For example, a nitrogen protecting group may be a group that is removed in vivo after administration to a host, in vitro by cells, or during the manufacturing process. Suitable nitrogen protecting groups useful in the present invention are described in Greene and Wuts in Protective Groups in Organic Synthesis (1991) New York, John Wiley and Sons, Inc.

[0144] As used herein, the term “oxygen protecting group” refers to a moiety that is covalently bonded to oxygen, is removable, and is typically substituted with hydrogen where appropriate. For example, an oxygen protecting group may be a group that is removed in vivo after administration to a host, in vitro by cells, or during the manufacturing process. Suitable oxygen protecting groups useful in the present invention are described in Greene and Wuts in Protective Groups in Organic Synthesis (1991) New York, John Wiley and Sons, Inc.

[0145] "Phosphate" refers to the group -OP(O)(OH)2.

[0146] Unless otherwise specified, "phosphate ester" refers to monophosphates, diphosphates, and triphosphates.

[0147] The terms "phosphoamidate," "phosphoramide," or "phosphoroamidate" refer to a moiety having three oxygen groups and phosphorus bonded to an amine (which may be substituted). Suitable phosphoramidates useful for the present invention are described by Madela, Karolina, and McGuigan in 2012. "Progress in the development of anti-hepatitis C virus nucleoside and nucleotide prodrugs", Future Medicinal Chemistry 4(5), pages 625-650 10:1021 / jm300074y, And described in Dominique, McGuigan and Balzarini in 2004, "Aryloxy Phosphoramidate Triesters as Pro-Tides", Mini Reviews in Medicinal Chemistry 4(4), pages 371-381. Further phosphoramidates useful for the present invention are U.S. Patents No. 5,233,031, No. 7,115,590, No. 7,547,704, No. 7,879,815, No. 7,888,330, No. 7,902,202, No. 7,951,789, No. 7,964,580, No. 8,071,568, No. 8,148,349, and No. 8,263 These are described in Patent Nos. 575, 8,324,179, 8,334,270, 8,552,021, 8,563,530, 8,580,765, 8,735,372, 8,759,318, European Patent No. 2120565, European Patent No. 1143995, U.S. Patent No. 6,455,513, and 8,334,270. Other phosphoramidates are described in the nucleoside patents described in the background art of the present invention.

[0148] The phosphoramidate groups used in this invention include those with the following structures. [ka]

[0149] Other phosphoramidates used in the present invention include those with the following structures. [ka] During the ceremony, RP1 This is a substituted linear, branched, or cyclic alkyl group, or a substituted aryl, heteroaryl, or heterocyclic group, or a linked combination thereof. R P2 -NR N1 R N2 It is a group or a B' group, During the ceremony, R N1 and R N2 These are H and C, which are independent of each other. 1~8 Alkyl, (C3-C7 cycloalkyl)C0-C4 alkyl, (aryl)C0-C4 alkyl-, (C3-C6 heterocyclo)C0-C4 alkyl-, or (heteroaryl)C0-C4 alkyl-, which may be substituted, or R N1 and R N2 It links with the nitrogen atom to which it is attached to form a 3- to 7-membered heterocycle. B' is, [ka] It is the basis, During the ceremony, R 16 This is a side chain of an amino acid, often selected from the group consisting of hydrogen, (C1-C8) alkyl, (C2-C8) alkenyl, (C2-C8) alkynyl, (C3-C8 cycloalkyl)C0-C4 alkyl-, (aryl)C0-C4 alkyl-, (C3-C6 heterocyclo)C0-C4 alkyl-, (heteroaryl)C0-C4 alkyl-, or an amino acid side chain, for example, (unless otherwise specified herein) alanine, β-alanine, arginine, asparagine, aspartic acid, cysteine, cystine, glutamic acid, glutamine, glycine, phenylalanine, histidine, isoleucine, lysine, leucine, methionine, proline, serine, threonine, valine, tryptophan, or tyrosine (often, R 16 (These are hydrogen, methyl, isopropyl, or isobutyl.) R 17This is a side chain of an amino acid, often selected from the group consisting of hydrogen, (C1-C8) alkyl, (C2-C8) alkenyl, (C2-C8) alkynyl, (C3-C8 cycloalkyl)C0-C4 alkyl-, (aryl)C0-C4 alkyl-, (C3-C6 heterocyclo)C0-C4 alkyl-, (heteroaryl)C0-C4 alkyl-, or an amino acid side chain, for example, (unless otherwise specified herein) alanine, β-alanine, arginine, asparagine, aspartic acid, cysteine, cystine, glutamic acid, glutamine, glycine, phenylalanine, histidine, isoleucine, lysine, leucine, methionine, proline, serine, threonine, valine, tryptophan, or tyrosine (often, R 17 (These are hydrogen, methyl, isopropyl, or isobutyl.) R 18 is hydrogen or a C1-C3 alkyl group, or R 16 and R 17 This may form a (C3-C7) cycloalkyl or (C3-C7) heterocyclic group. Or, R 18 and R 16 Or R 17 (C3~C6) heterocyclic groups may be formed. R 19 These are hydrogen, (C1~C6)alkyl, (C3~C6)alkenyl, (C3~C6)alkynyl, (C3~C8cycloalkyl)C0~C4alkyl-, (aryl)C0~C4alkyl-, (C3~C6 heterocyclo)C0~C4alkyl-, and (heteroaryl)C0~C4alkyl-. Alternatively, B' is, [ka] It is the basis, During the ceremony, R 20 These are hydrogen, (C1-C3) alkyl, (C3-C8 cycloalkyl)C0-C4 alkyl-, (aryl)C0-C4 alkyl-, (C3-C6 heterocyclo)C0-C4 alkyl-, or (heteroaryl)C0-C4 alkyl-. R21 These are hydrogen, (C1-C3) alkyl, (C3-C8 cycloalkyl)C0-C4 alkyl-, (aryl)C0-C4 alkyl-, (C3-C6 heterocyclo)C0-C4 alkyl-, or (heteroaryl)C0-C4 alkyl-. R 18 and R 19 It is defined above.

[0150] Preferred R P1 Examples of the groups include substituted phenyl groups, naphthyl groups, and monocyclic heteroaryl groups, particularly groups that enhance the bioavailability of the compound in patient cells (especially lipophilic groups) and exhibit reduced toxicity, enhanced therapeutic index, and enhanced pharmacokinetics (the compound is metabolized and excreted more slowly).

[0151] The term phosphoramidate is used throughout this specification to describe a group found at the 5' or 3' position of the furanose ring of a nucleoside compound that forms a prodrug form of the nucleoside compound. In one embodiment, the phosphoramidate is found at both the 5' and 3' positions of the furanose ring of a nucleoside compound and can form a prodrug form of the nucleoside compound. In another embodiment, the phosphoramidate found at the 5' position of the furanose ring of a nucleoside can form a cyclic phosphoramidate compound by forming a bond with the 3'-hydroxyl substituent at the 3' position of the furanose ring of the nucleoside compound, thereby forming a prodrug form of the nucleoside compound.

[0152] The terms "thiophosphoamidate," "thiophosphoramide," or "thiophosphoroamidate" refer to a moiety having a sulfur-bonded phosphorus, two oxygen groups, and an amine (which may be substituted). Thiophosphoramides useful in the present invention are described in U.S. Patent No. 8,772,474 and International Publication No. 2012 / 040124.

[0153] The thiophosphoramide groups used in this invention include those with the following structures. [ka]

[0154] Other thiophosphoramidates include those with the following structure. [ka] During the ceremony, R P1 This is a substituted linear, branched, or cyclic alkyl group, or a substituted aryl, heteroaryl, or heterocyclic group, or a linked combination thereof. R P2 -NR N1 R N2 It is a group or a B' group, During the ceremony, R N1 and R N2 These are H and C, which are independent of each other. 1~ C8 alkyl, (C3-C7 cycloalkyl)C0-C4 alkyl-, (aryl)C0-C4 alkyl-, (C3-C6 heterocyclo)C0-C4 alkyl-, or (heteroaryl)C0-C4 alkyl-, or R N1 and R N2 It links with the nitrogen atom to which it is attached to form a 3- to 7-membered heterocycle. B' is, [ka] It is the basis, During the ceremony, R 16This is a side chain of an amino acid, often selected from the group consisting of hydrogen, (C1-C8) alkyl, (C2-C8) alkenyl, (C2-C8) alkynyl, (C3-C8 cycloalkyl)C0-C4 alkyl-, (aryl)C0-C4 alkyl-, (C3-C6 heterocyclo)C0-C4 alkyl-, (heteroaryl)C0-C4 alkyl-, or an amino acid side chain, for example, (unless otherwise specified herein) alanine, β-alanine, arginine, asparagine, aspartic acid, cysteine, cystine, glutamic acid, glutamine, glycine, phenylalanine, histidine, isoleucine, lysine, leucine, methionine, proline, serine, threonine, valine, tryptophan, or tyrosine (often, R 16 (These are hydrogen, methyl, isopropyl, or isobutyl.) R 17 This is a side chain of an amino acid, often selected from the group consisting of hydrogen, (C1-C8) alkyl, (C2-C8) alkenyl, (C2-C8) alkynyl, (C3-C8 cycloalkyl)C0-C4 alkyl-, (aryl)C0-C4 alkyl-, (C3-C6 heterocyclo)C0-C4 alkyl-, (heteroaryl)C0-C4 alkyl-, or an amino acid side chain, for example, (unless otherwise specified herein) alanine, β-alanine, arginine, asparagine, aspartic acid, cysteine, cystine, glutamic acid, glutamine, glycine, phenylalanine, histidine, isoleucine, lysine, leucine, methionine, proline, serine, threonine, valine, tryptophan, or tyrosine (often, R 17 (These are hydrogen, methyl, isopropyl, or isobutyl.) R 18 is hydrogen or a C1-C3 alkyl group, or R 16 and R 17 This may form a (C3-C7) cycloalkyl or (C3-C7) heterocyclic group. Or, R 18 and R 16 Or R 17 (C3~C6) heterocyclic groups may be formed. R 19These are hydrogen, (C1~C6)alkyl, (C3~C6)alkenyl, (C3~C6)alkynyl, (C3~C8cycloalkyl)C0~C4alkyl-, (aryl)C0~C4alkyl-, (C3~C6 heterocyclo)C0~C4alkyl-, and (heteroaryl)C0~C4alkyl-. Alternatively, B' is, [ka] It is the basis, R 18 , R 19 , R 20 and R 21 It is defined as above.

[0155] Preferred R P1 Examples of the groups include substituted phenyl groups, naphthyl groups, and monocyclic heteroaryl groups, particularly groups that enhance the bioavailability of the compound in patient cells (especially lipophilic groups) and exhibit reduced toxicity, enhanced therapeutic index, and enhanced pharmacokinetics (the compound is metabolized and excreted more slowly).

[0156] Thiophosphoramides are found at the 5' or 3' position of the furanose ring of a nucleoside compound and can form a prodrug form of the nucleoside compound. In one embodiment, thiophosphoramides are found at both the 5' and 3' positions of the furanose ring of a nucleoside compound and can form a prodrug form of the nucleoside compound. In another embodiment, thiophosphoramides found at the 5' position of the furanose ring of a nucleoside can form a cyclic thiophosphoramide compound by forming a bond with the 3'-hydroxyl substituent at the 3' position of the furanose ring of the nucleoside compound and forming a prodrug form of the nucleoside compound.

[0157] As used in the context of this invention, the term "D configuration" refers to a non-natural nucleoside or a principle configuration that mimics the natural configuration of the sugar moiety, in contrast to the "L" configuration. The terms "β" or "β-anomer" are used in reference to nucleoside analogs in which a nucleoside base is formed (positioned) on the plane of the furanose moiety in the nucleoside analog.

[0158] The terms “simultaneous administration” and “concurrent administration,” or “combination therapy,” are used to describe the administration of at least one 2'-deoxy-2'-α-fluoro-2'-β-C-nucleoside compound according to the present invention in combination with at least one additional anti-HCV agent, for example, at least one other activating agent, which, where appropriate, includes other 2'-deoxy-2'-α-fluoro-2'-β-C-nucleoside agents disclosed herein. The timing of simultaneous administration is best determined by the specialist treating the patient. Sometimes, it is preferable for the drugs to be administered simultaneously. Alternatively, the drugs selected for combination therapy may be administered to the patient at different times. Of course, if two or more viruses or other infections or other medical conditions are present, the compounds of the present invention may be combined with other agents as needed to treat the other infections or medical conditions.

[0159] As used herein, the term “host” refers to a single-celled or multicellular organism, typically human, including cell lines and animals, in which the HCV virus can replicate. Specifically, the term host refers to infected cells, cells transfected with all or part of the HCV genome, and animals, particularly primates (including chimpanzees) and humans. In most animal applications of the present invention, the host is a human patient. However, veterinary applications, in certain indications, are clearly anticipated by the present invention (e.g., chimpanzees). The host may also be, for example, a cattle, horse, bird, dog, cat, etc.

[0160] Isotope substitution The present invention includes compounds and the use of compounds involving the substitution of an atom with a desired isotope in an amount exceeding the natural abundance, i.e., a concentrated amount of the isotope. An isotope is an atom that has the same atomic number but different mass numbers, i.e., the same number of protons but different numbers of neutrons. A general example, and not limited to, is deuterium, which is an isotope of hydrogen. 2 H) and tritium ( 3 H) may be used at any location in the described structure. Alternatively or additionally, a carbon isotope, for example, 13 C and 14 C may be used. A preferred isotope substitution is the substitution of one or more hydrogen atoms with deuterium at one or more positions on the molecule to improve the performance of the drug. Deuterium may be bound at the cleavage site during metabolism (α-deuterium rate isotope effect) or adjacent to or near the cleavage site (β-deuterium rate isotope effect). Achillion Pharmaceuticals, Inc. (International Public Distributor) Publication No. 2014 / 169278 and International Publication No. 2014 / 169280 describe the deuteration of nucleotides, including the 5th position of molecules, to improve their pharmacokinetics or efficacy.

[0161] Substitution with isotopes such as deuterium can provide specific therapeutic benefits due to greater metabolic stability, such as increased in vivo half-life or reduced required dosage. Substitution of hydrogen with deuterium at sites of metabolic degradation can reduce the rate of metabolism or eliminate metabolism at its binding site. At any position in a compound where a hydrogen atom may be present, the hydrogen atom can be replaced with protium ( 1 H), deuterium ( 2 H) and tritium ( 3 It may be any hydrogen isotope containing H). Therefore, unless otherwise explicitly indicated in the context, references to compounds herein encompass all possible forms of isotopes.

[0162] The term "isotope-labeled" analog is "deuterized analog".13 "C-labeled analog" or "deuterized / 13 This refers to an analog that is a "C-labeled analog." The term "deuterated analog" refers to the compounds, H-isotopes, i.e., hydrogen / protium ( 1 H) is an H-isotope, that is, deuterium ( 2 This means substitution by H). Deuterium substitution may be partial or complete. Partial deuterium substitution means that at least one hydrogen is substituted by at least one deuterium. In certain embodiments, the isotope is concentrated to 90%, 95%, or 99% or more in the isotope at any desired position. In some embodiments, it is deuterium that is concentrated to 90%, 95%, or 99% at the desired position. Unless otherwise specified, deuteration is at least 80% at the selected position. Deuteration of the nucleoside may occur at any substituted hydrogen that provides the desired result.

[0163] III. Methods of treatment or prevention The treatments used herein refer to the administration of active compounds to a host infected with the HCV virus.

[0164] The terms “prophylactic” or “preventative,” when used, refer to the administration of an active compound to prevent or reduce the likelihood of viral injury. The present invention includes both therapeutic and prophylactic or preventative therapies. In one embodiment, the active compound is administered to a host who has been exposed to the hepatitis C virus and is therefore at risk of infection. The present invention relates to hepatitis C virus, including drug-resistant and multidrug-resistant forms of HCV, and to disease states, conditions, or complications associated with HCV infection, including cirrhosis and associated hepatotoxicity, as well as, in particular, other secondary conditions of HCV infection, such as weakness, loss of appetite, weight loss, breast enlargement (especially in men), rash (especially of the palms), blood clotting problems, spider angiography of the skin, confusion, coma (encephalopathy), fluid accumulation in the abdominal cavity (ascites), esophageal varices, portal hypertension, renal failure, splenomegaly, cytopenia, anemia, thrombocytopenia, jaundice, and hepatocellular carcinoma. The above method relates to an effective amount of at least one β-D-2'-D-2'-α-fluoro-2'-β-C-substituted-2-modified-N as described herein. 6 - The method comprises administering the substituted purine nucleotide to a host in need, optionally in combination with at least one additional bioactive agent, such as an additional anti-HCV agent, and further in combination with pharmaceutically acceptable carrier additives and / or excipients.

[0165] In yet another aspect, the present invention provides treatment for HCV infection, or in particular for related disease conditions or subsequent disease conditions, symptoms, or complications, including cirrhosis and associated hepatotoxicity, weakness, loss of appetite, weight loss, breast enlargement (especially in men), rash (especially on the palms), blood clotting problems, spider angiography of the skin, confusion, coma (encephalopathy), fluid accumulation in the abdominal cavity (ascites), esophageal varices, portal hypertension, renal failure, splenomegaly, cytopenia, anemia, thrombocytopenia, jaundice, and hepatocellular carcinoma. A method for stopping or preventing the disease, the above method involves administering to a patient at risk at least one compound according to the present invention, as described above in an effective amount, in combination with a pharmaceutically acceptable carrier, additive or excipient, and optionally in combination with another anti-HCV agent. In another embodiment, the active compound of the present invention may be administered to a patient to protect a new organ after a liver transplant associated with hepatitis.

[0166] If desired, the parent compound can be provided directly or indirectly by administration to a recipient, or as any salt or prodrug that exhibits activity itself, such as 5'-stabilized β-D-2'-D-2'-α-fluoro-2'-β-C-substituted-2-modified-N 6 - Substituted purine nucleotides can be administered. Non-limiting examples include pharmaceutically acceptable salts and compounds modified with functional groups such as hydroxyl or amine functional groups to modify the biological activity, pharmacokinetics, half-life, controlled delivery, lipophilicity, absorption rate, ease of phosphorylation to active 5'-triphosphate, or efficiency of delivery using a desired administration route. Methods for modifying the properties of active compounds to achieve target properties are known to those skilled in the art or can be readily determined by standard methods, such as acylation, phosphorylation, thiophosphoramidation, phosphoramidation, phosphononation, alkylation, or pegylation.

[0167] IV. Pharmaceutical Compositions In aspects of the present invention, the pharmaceutical composition according to the present invention may optionally be combined with a pharmaceutically acceptable carrier, additive or excipient, and further optionally with at least one other active compound, or alternately with at least one 5'-stabilized β-D-2'-D-2'-α-fluoro-2'-β-C-substituted-2-modified-N compound described herein in an amount effective against the HCV virus. 6 -Contains substituted purine nucleotide compounds.

[0168] In aspects of the present invention, the pharmaceutical composition according to the present invention may optionally be combined with a pharmaceutically acceptable carrier, additive or excipient, and further optionally with at least one other antiviral agent such as an anti-HCV agent, to contain an amount effective against HCV of at least one active β-D-2'-D-2'-α-fluoro-2'-β-C-substituted-2-modified-N as described herein. 6 -Contains substituted purine nucleotide compounds.

[0169] The present invention provides an effective amount of the β-D-2'-D-2'-α-fluoro-2'-β-C-substituted-2-modified-N to treat hepatitis C virus infection. 6 -The present invention comprises a pharmaceutical composition containing one of the substituted purine nucleotide compounds, or a salt or prodrug thereof, in a pharmaceutically acceptable carrier or excipient. In an alternative embodiment, the present invention provides an amount of the β-D-2'-D-2'-α-fluoro-2'-β-C-substituted-2-modified-N of the present invention that is effective in preventing hepatitis C virus infection. 6 -The pharmaceutical composition comprises one of the substituted purine nucleotide compounds, or a salt or prodrug thereof, in a pharmaceutically acceptable carrier or excipient.

[0170] Those skilled in the art will recognize that the therapeutically effective dose varies depending on the infection or condition being treated, its severity, the treatment plan employed, the pharmacokinetics of the drug used, and also on the patient or subject (animal or human) being treated, and that such a dose can be determined by the attending physician or specialist.

[0171] The present invention provides 5'-stabilized β-D-2'-D-2'-α-fluoro-2'-β-C-substitution-2-modification-N 6- Substituted purine nucleotide compounds may be formulated into mixtures with pharmaceutically acceptable carriers. Generally, it is preferable to administer pharmaceutical compositions in oral dosage forms, but certain formulations may be administered by other routes, including parenteral, intravenous, intramuscular, topical, transdermal, buccal, subcutaneous, suppositories, and nasal sprays. Intravenous and intramuscular formulations are often administered in sterile saline. Those skilled in the art may modify formulations to make them soluble in water or other vehicles, for example, if this is done with sufficient common knowledge of the art. This can be easily accomplished with minor modifications (such as salt formulations or esterification). Furthermore, modifying the administration route and drug regimen of specific compounds to control the pharmacokinetics of the compounds of the present invention for the greatest beneficial effect in the patient is also part of the usual practice for those skilled in the art.

[0172] In certain pharmaceutical dosage forms, prodrug forms of the compounds are particularly preferred, including acylated (acetylated or otherwise) and ether (alkyl and related) derivatives, phosphate esters, thiophosphoramides, phosphoramides, and various salt forms of the compounds of the present invention. Those skilled in the art will recognize how readily the compounds of the present invention can be converted into prodrug forms to facilitate delivery of the active compound to the target site of the host organism or patient. Where applicable, the advantageous pharmacokinetic parameters of the prodrug form should be utilized in the delivery of the active compound to the target site of the host organism or patient to maximize the intended effect of the compounds.

[0173] The amount of compound contained in the therapeutically active formulation according to the present invention is an amount effective for treating HCV infection, reducing the likelihood of HCV infection, inhibiting, reducing, and / or eliminating HCV, or for its secondary effects, including disease states, conditions, and / or complications that occur following HCV. Generally, the therapeutically effective amount of the compound of the present invention in a pharmaceutical dosage form is usually in the range of about 0.001 mg / kg to about 100 mg / kg or more per day to a patient, depending on the compound used, the condition or infection being treated, and the route of administration, and is often slightly less than about 0.1 mg / kg to considerably more than about 25 mg / kg per day. The active nucleoside compound according to the present invention is often administered to a patient in an amount ranging from about 0.1 mg / kg to about 15 mg / kg per day, depending on the pharmacokinetics of the drug in the patient. This dosage range generally results in effective blood concentrations of the active compound, which can range from approximately 0.001 micrograms to 100 micrograms per cubic centimeter of the patient's blood, or from approximately 0.05 micrograms to 100 micrograms.

[0174] Often, to treat, prevent or delay the onset of these infections, and / or to reduce the likelihood of HCV virus infection or secondary disease states, conditions, or complications of HCV, the compositions are administered orally at least once daily in amounts ranging from about 250 micrograms to a maximum of about 500 mg or more, for example, up to four times daily in amounts of at least 25 milligrams, 50 milligrams, 100 milligrams, 150 milligrams, 250 milligrams, or 500 milligrams. Although the compounds of the present invention are often administered orally, they may also be administered parenterally, topically, or in suppository form, and intranasally as nasal drops or otherwise as described herein.

[0175] In the case of co-administration of the compounds of the present invention in combination with other anti-HCV compounds as described herein, the amount of the compound according to the present invention administered is considerably greater than about 0.01 mg / kg to about 500 mg / kg to the patient, depending on the second agent administered co-administered, as well as the efficacy against the virus, the patient's condition, the severity of the disease or infection being treated, and the route of administration. Other anti-HCV agents may be administered in amounts ranging, for example, from about 0.01 mg / kg to about 500 mg / kg. In certain preferred embodiments, these compounds may generally be administered in amounts ranging from about 0.5 mg / kg to about 50 mg / kg or more (usually up to about 100 mg / kg), depending on the pharmacokinetics of the two agents in the patient. These dosage ranges generally result in effective blood concentrations of the active compound in the patient.

[0176] With regard to the object of the present invention, the prophylactic or inhibitory effective amount of the composition according to the present invention falls within the same concentration range as the therapeutic effective amount described above, and is usually the same as the therapeutic effective amount.

[0177] The active compound is administered continuously (intravenous infusion) or several times a day orally or nasally. Administration may include oral administration (e.g., QID) or transdermal administration, as well as other routes of administration, including oral, topical, parenteral, intramuscular, intravenous, subcutaneous, transdermal (may include penetration enhancers), buccal, and suppository administration. Enteric-coated oral tablets may also be used to enhance the bioavailability of the compound for oral administration. The most effective dosage form depends on the bioavailability / pharmacokinetics of the specific drug selected, as well as the severity of the patient's disease. Oral dosage forms are particularly preferred due to their ease of administration and the expected favorable patient compliance.

[0178] To prepare the pharmaceutical compositions according to the present invention, a therapeutically effective amount of one or more compounds according to the present invention is often essentially mixed with a pharmaceutically acceptable carrier in accordance with conventional pharmaceutical formulation techniques to produce a dosage. The carrier may take various forms depending on the formulation preferred for oral or parenteral administration, for example. Any of the usual pharmaceutical media may be used in the preparation of the pharmaceutical compositions into oral dosage forms. Therefore, suitable carriers and additives including water, glycol, oil, alcohol, fragrance, preservative, colorant, etc. may be used for liquid oral formulations such as suspensions, elixirs, and solutions. Suitable carriers and additives including starch, sugar carriers such as dextrose, manifolds, lactose, and related carriers, diluents, granulators, lubricants, binders, disintegrants, etc. may be used for solid oral formulations such as powders, tablets, capsules, and suppositories.

[0179] For parenteral formulations, the carrier usually contains sterile water or an aqueous sodium chloride solution, but may also contain other components, including those that assist in dispersion. If sterile water is used and maintained as sterile, the composition and carrier must, of course, also be sterilized. Alternatively, an injection suspension may be prepared, in which case an appropriate liquid carrier, suspending agent, etc., may be used.

[0180] Furthermore, liposome suspensions (containing liposomes targeted to viral antigens) may be prepared by conventional methods to produce pharmaceutically acceptable carriers. This may be suitable for the delivery of the nucleoside compound according to the present invention in the form of a free nucleoside, acyl / alkyl nucleoside, or phosphate ester prodrug.

[0181] In a typical embodiment of the present invention, the compounds and compositions described above are used to treat, prevent, or delay HCV infection or secondary disease conditions, symptoms, or complications of HCV.

[0182] V. Combination therapy and alternating therapy It is well recognized that drug-resistant variants of the virus may emerge after prolonged treatment with antiviral agents. Drug resistance most typically results from mutations in the genes encoding enzymes used in viral replication. The efficacy of a drug against HCV infection may be prolonged, increased, or restored by combining it with another (perhaps two or three more) antiviral compound that induces different mutations or acts through a different pathway than that of the principal drug, or by administering the aforementioned compound alternately with the antiviral agent. Alternatively, pharmacokinetics, distribution in the body, half-life, or other parameters of the drug may be modified by such combination therapy (which may include alternating therapy, if considered to be synergistic). Disclosed β-D-2'-D-2'-α-fluoro-2'-β-C-substitution-2-modification-N 6 - Substituted purine nucleotides are NS5B polymerase inhibitors, and it may be useful to administer the above compounds to the host in combination with, for example, the following: (1) Protease inhibitors (NS3 / 4A protease inhibitors, etc.) (2) NS5A inhibitors, (3) Another NS5B polymerase inhibitor, (4) NS5B non-substrate inhibitors, (5) Interferon alpha-2a, which may be pegylated or otherwise modified, and / or ribavirin (6) Non-substrate inhibitors, (7) Helicase inhibitors, (8) Antisense oligodeoxynucleotide (S-ODN), (9) Aptamer, (10) Nuclease-resistant ribozymes, (11) iRNAs including microRNA and siRNA, (12) Antibodies against the virus, partial antibodies, or domain antibodies, (13) A viral antigen or partial antigen that induces a host antibody response.

[0183] The present invention β-D-2'-D-2'-α-fluoro-2'-β-C-substitution-2-modification-N 6 -The following are some non-exclusive examples of anti-HCV agents that can be administered in combination with substituted purine nucleotides: (i) Protease inhibitors such as telaprevir (Insivec®), boceprevir (Victorelis®), simeprevir (Olisio®), paritaprevir (ABT-450), ACH-2684, AZD-7295, BMS-791325, danoprevir, filibuvir, GS-9256, GS-9451, MK-5172, cetrobuvir, sobaprevir, tegobuvir, VX-135, VX-222 and AL-220, (ii) NS5A inhibitors such as ACH-2928, ACH-3102, IDX-719, daclatasvir, ledipasvir, and ombitasvir (ABT-267) (iii) NS5B inhibitors such as ACH-3422, AZD-7295, cremizole, ITX-5061, PPI-461, PPI-688, Sovaldi (trademark), MK-3682, and mericitabine. (iv) NS5B inhibitors such as ABT-333 and MBX-700, (v) Antibodies such as GS-6624.

[0184] β-D-2'-D-2'-α-fluoro-2'-β-C-substituted-2-modified-N 6 -When a substituted purine nucleotide is administered to treat severe hepatitis C virus linked to liver cancer or cirrhosis, in one embodiment, for example, Andrew Zhu in "New Agents on "The Horizon in Hepatocellular Carcinoma," Therapeutic Advances in Medical Oncology, V 5(1), (January 2013), 41-50, describes the treatment of hepatocellular carcinoma (HCC). The above compounds can be administered in combination with, or alternately with, other drugs typically used for HCC. Examples of compounds suitable for combination therapy when the host has or is at risk of having HCC include anti-angiogenic agents, sunitinib, brivanib, linifanib, ramucirumab, bevacizumab, cedilanib, pazopanib, TSU-68, lenvatinib, antibodies against EGFR, mTor inhibitors, MEK inhibitors, and histone deacetylase inhibitors.

[0185] The drugs currently approved for influenza are amantadine, rimantadine, and oseltamivir. Any of these drugs can be used in combination with, or alternately with, the active compounds presented herein to treat viral infections susceptible to them. Ribavirin is used to treat measles, influenza A, influenza B, parainfluenza, severe RSV bronchiolitis, and SARS, as well as other viral infections, and is therefore particularly useful when combined with the compounds of the present invention for the treatment of hosts infected with single-stranded RNA viruses. Palivizumab is approved for use in infants at high risk of RSV infection.

[0186] Currently, there are no approved drugs for West Nile virus. Doctors recommend hospitalization, intravenous fluids, It is recommended to provide intensive supportive care, which may include the use of a ventilator to assist breathing, medications to manage seizures, antibiotics to treat brain swelling, nausea and vomiting, and antibiotics to prevent bacterial infections and further exacerbation of the disease. This highlights the importance of the compounds of the present invention for viral drug therapy.

[0187] VI. The present invention of β-D-2'-D-2'-α-fluoro-2'-β-C-substitution-2-modification-N 6 - Process of producing substituted purine nucleotides General methods for providing the compounds of the present invention are known in the art or are described herein. The synthesis of 2'-chloronucleotides is described in U.S. Patent Application Publication No. 20150366888, International Publication No. 2014058801, International Publication No. 2015 / 066370, and International Publication No. 2015200219.

[0188] The following abbreviations are used in the synthesis mechanism. CBr4: Carbon tetrabromide DBU:1,8-Diazabicyclo[5.4.0]Undeca-7-En DCM: Dichloromethane THF: Tetrahydrofuran (THF), anhydrous æ:ethyl acetate EtOH: Ethanol Li(OtBu)3AlH: Lithium hydride tri-tert-butoxyaluminum hydride Na2SO4: Sodium sulfate (anhydrous) MeCN: Acetonitrile MeNH2: Methylamine MeOH: methanol Na2SO4: Sodium sulfate NaHCO3: Sodium Bicarbonate NH4Cl: Ammonium chloride NH4OH: Ammonium hydroxide PE: Petroleum ether Ph3P: Triphenylphosphine Silica gel (230-400 mesh, Sorbent) t-BuMgCl: t-Butyl magnesium chloride t-BuOK: Sodium tert-butoxide t-BuOH:tert-butanol [Examples]

[0189] general law 1 H, 19 F and 31The NMR spectrum of P was recorded on a 300 MHz Fourier transform Bruker spectrometer. Spectra were obtained from samples prepared in 5 mm diameter tubes in CDCl3, CD3OD, or DMSO-d6. Spin multiplicity is indicated by the symbols s (singlet), d (doublet), t (triplet), m (multinumber), and br (broad). Coupling constants (J) are reported in Hz. MS spectra were obtained using electrospray ionization (ESI) on an Agilent Technologies 6120 quadrupole MS instrument. The above reaction was carried out using a dry solvent, typically Sigma-Aldrich anhydrous solvent. The experiment was conducted under a nitrogen atmosphere. All common chemicals were purchased from commercial suppliers. [ka] i) Li(OtBu)3AlH, THF, -30℃→-15℃; ii) PPh3, CBr4, DCM, -20℃→0℃; iii) 2-amino-6-chloropurine, tBuOK, tBuOH / MeCN 9:1, 65℃; iv) MeNH2 (33%), MeOH, 85℃; v) Isopropyl((R,S)-(pentafluorophenoxy)-phenoxy-phosphoryl)-L-alaninate, tBuMgCl, THF, 0℃→room temperature.

[0190] Example 1. Preparation of isopropyl((((R,S)-(2R,3R,4R,5R)-5-(2-amino-6-(methylamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate

[0191] Step 1. Preparation of ((2R,3R,4R,5R)-3-(benzoyloxy)-5-bromo-4-fluoro-4-methyltetrahydrofuran-2-yl)methylbenzoate (2). To a solution of (2R)-3,5-di-O-benzoyl-2-fluoro-2-C-methyl-D-ribono-γ-lactone (24.8 g, 66.6 mmol) in dry THF (333 mL), lithium tri-tert-butoxyaluminum hydride (1.0 M in THF, 22.6 mL, 22.6 mmol) was added dropwise under a nitrogen atmosphere, cooled to -30°C. After the addition was complete, the reaction mixture was slowly warmed to -15°C over 90 minutes, then ethyl acetate (300 mL) was added, and the mixture was quenched with saturated NH4Cl aqueous solution (200 mL). The resulting solution was filtered over Celite®, and the filtrate was extracted twice with ethyl acetate. The combined organic matter was dried (Na2SO4), filtered, and concentrated. The residue was transferred to a dry DCM (225 mL) under a nitrogen atmosphere and cooled to -20°C. PPh3 (19.1 g, 72.8 mmol) was then added. After stirring at -20°C for 10 minutes, CBr4 (26.0 g, 78.4 mmol) was added, and the reaction mixture was slowly warmed to 0°C over 2 hours. The resulting mixture was poured onto a silica gel column and eluted with PE / siRNA (gradient from 100:0 to 80:20). The fraction containing α-bromofuranosides was collected and concentrated to obtain Product 2 (18.1 g, 41.3 mmol, 62% over two steps) as a highly viscous, colorless oil. 1 H NMR(300MHz,CDCl3)δ 8.15~8.11(m,2H), 8.04~8.01(m,2H), 7.64~7.55(m,2H), 7.51~7.41(m,4H), 6.34(d,J=1.6Hz,1H), 5.29(dd,J=5.5 ,3.1Hz,1H), 4.89~4.85(m,1H), 4.78(dd,J=12.5,3.2Hz,1H), 4.63(dd,J=12.5,4.5Hz,1H), 1.72(d,J=21.6Hz,3H). 19 F NMR(282MHz, CDCl3)δ -150.0.

[0192] Step 2. Preparation of (2R,3R,4R,5R)-5-(2-amino-6-chloro-9H-purine-9-yl)-2-(benzoyloxymethyl)-4-fluoro-4-methyltetrahydrofuran-3-ylbenzoate (3). 2-amino-6-chloropurine (2.63 g, 15.5 mmol) was suspended in t-BuOH (54 mL) under a nitrogen atmosphere. The reaction mixture was heated to 30°C, and then potassium tert-butoxide (1.69 g, 15.1 mmol) was added. After 45 minutes, a solution of bromofuranoside 2 (2.24 g, 5.12 mmol) dissolved in anhydrous MeCN (6 mL) was added, and the reaction mixture was heated to 65°C for 16 hours, after which it was cooled to room temperature. Saturated NH4Cl aqueous solution (70 mL) was added, and the resulting solution was extracted with ELISA (60 mL three times). The combined organic matter was dried (Na2SO4), filtered, and concentrated. The residue was purified twice by column chromatography (PE / siRNA gradient from 80:20 to 0:100, followed by a gradient from 60:40 to 20:80) to obtain Product 3 (1.56 g, 2.96 mmol, 57%) as a white solid. 1 H NMR(300MHz,CDCl3)δ 8.05~8.02(m,2H), 7.95~7.92(m,2H), 7.88(s,1H), 7.63~7.57(m,1H), 7.53~7.41(m,3H), 7.35~7.30(m,2H), 6.43(dd,J=22.6,9.1Hz,1H), 6 .12(d,J=18.3Hz,1H), 5.34(brs,2H), 5.00(dd,J=11.9,4.5Hz,1H), 4.79~4.73(m,1H), 4.60(dd,J=11.9,5.3Hz,1H), 1.34(d,J=22.6Hz,3H). 19 F NMR(282MHz, CDCl3)δ -157.0. C 25 H 22 FN5O5[M+H] + The calculated MS(ESI)m / z value was 526.9; the measured value was 527.0.

[0193] Step 3. Preparation of (2R,3R,4R,5R)-5-(2-amino-6-(methylamino)-9H-purine-9-yl)-4-fluoro-2-(hydroxymethyl)-4-methyltetrahydrofuran-3-ol (4). Methylamine (33% in anhydrous EtOH, 1.7 mL, 1.81 mmol) was added to a solution of compound 3 (575 mg, 1.09 mmol) in MeOH (9 mL). The reaction mixture was heated to 85°C in a sealed tube for 16 hours, cooled to room temperature, and concentrated. The residue was purified by column chromatography (DCM / MeOH gradient from 100:0 to 85:15) followed by reversed-phase column chromatography (H2O / MeOH gradient from 100:0 to 0:100) to obtain product 4 (286 mg, 0.91 mmol, 84%) as a white solid. 1 H NMR(300MHz,CD3OD)δ 8.06(s,1H), 6.11(d,J=18.1Hz,1H), 4.41(dd,J=24.4,9.1Hz,1H), 4.07~4.01 (m,2H), 3.86(dd,J=12.9,3.3Hz,1H), 3.04(brs,3H), 1.16(d,J=22.3Hz,3H). 19 F NMR(282MHz,CD3OD)δ -163.7. C 12 H 19 FN6O3[M+H] + The calculated MS(ESI)m / z value was 313.1; the measured value was 313.2.

[0194] Step 4. Preparation of isopropyl((((R,S)-(2R,3R,4R,5R)-5-(2-amino-6-(methylamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate (5). To a solution of compound 4 (114 mg, 365 μmol) in dry THF (4 mL), t-butyl magnesium chloride (1.0 M in THF, 0.66 mL, 660 μmol) was added over 10 minutes under a nitrogen atmosphere while cooled to 0°C. The reaction mixture was stirred at 0°C for 15 minutes, and then stirred at room temperature for another 15 minutes. After the reaction mixture was cooled to 0°C, a solution of isopropyl ((R,S)-(pentafluorophenoxy)-phenoxy-phosphoryl)-L-alaninate dissolved in dry THF (1 mL) was used. Ross, BS, Reddy, PG, Zhang, HR, Rachak Onda, S., and Sofia, MJ, J. Org, Chem., (2011), (253 mg, 558 μmol) was added dropwise over 10 minutes. The reaction mixture was stirred at 0°C for 30 minutes, then stirred at room temperature for 18 hours, and subsequently quenched with saturated NH4Cl aqueous solution (4 mL) and extracted with ELISA (5 mL three times). The combined organic matter was dried and filtered to (Na2SO4) and concentrated. The residue was purified by column chromatography (DCM / MeOH gradient from 100:0 to 90:10) followed by reverse-phase column chromatography (H2O / MeOH gradient from 100:0 to 0:100) to obtain product 5 (a mixture of diastereomers, 101 mg, 174 μmol, 48%) as a white solid. 1 H NMR(300MHz,CD3OD)δ 7.83(s,0.55H), 7.82(s,0.45H), 7.38~7.16(m,5H), 6.15(d,J=18.5Hz,0.45H), (d,J=18.8Hz,0.55H), 4.99~4.88(H2O and double, m, 1H), 4.65-4.36 (m, 3H), 4.25-4.17 (m, 1H), 3.97-3.85 (m, 1H), 3.05 (brs, 3H), 1.32-1.28 (m, 3H), 1.25-1.15 (m, 9H). 19 F NMR (282MHz, CD3OD) δ -162.8(s), -163.3(s). 31 PNMR(121MHz,CD3OD)δ 4.10(s), 3.99(s). C 24 H 34 FN7O7P[M+H] + MS(ESI)m / z calculated value: 582.2; measured value: 582.2. [ka] i) Me2NH:HCl, DBU, MeOH, 85℃; v) Isopropyl((R,S)-(pentafluorophenoxy)-phenoxy-phosphoryl)-L-alaninate, tBuMgCl, THF, 0℃.

[0195] Example 2. Preparation of isopropyl((((R,S)-(2R,3R,4R,5R)-5-(2-amino-6-(dimethylamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate (7).

[0196] Step 1. Preparation of (2R,3R,4R,5R)-5-(2-amino-6-(dimethylamino)-9H-purine-9-yl)-4-fluoro-2-(hydroxymethyl)-4-methyltetrahydrofuran-3-ol (6). To a solution of compound 3 (500 mg, 0.95 mmol) from Example 1 in MeOH (6 mL), dimethylamine hydrochloride (783 mg, 9.6 mmol) and 1,8-diazabicyclo[5.4.0]undeca-7-ene (1.43 mL, 9.6 mmol) were added. The reaction mixture was heated at 85 °C in a sealed tube for 6 hours, cooled to room temperature, and concentrated. The residue was purified by column chromatography (DCM / MeOH gradient from 100:0 to 85:15) followed by reverse-phase column chromatography (H2O / MeOH gradient from 100:0 to 0:100) to obtain product 6 (200 mg, 0.61 mmol, 64%) as a white solid. I obtained it. 1 H NMR(300MHz,CD3OD)δ 8.07(s,1H), 6.14(d,J=18.1Hz,1H), 4.41(dd,J=24.4,9.2Hz,1H), 4.08~4.02 (m,2H), 3.87(dd,J=12.8,2.9Hz,1H), 3.42(brs,6H), 1.16(d,J=22.0Hz,3H). 19 F NMR(282MHz,CD3OD)δ -163.8. C 13 H 20 FN6O3[M+H] + The calculated MS(ESI)m / z value is 327.2; the measured value is 327.2.

[0197] Step 2. Preparation of isopropyl((((R,S)-(2R,3R,4R,5R)-5-(2-amino-6-(dimethylamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate (7). To a solution of compound 6 (80 mg, 245 μmol) in dry THF (4 mL), tert-butylmagnesium chloride (1.0 M in THF, 0.64 mL, 640 μmol) was added dropwise over 10 minutes under a nitrogen atmosphere and cooled to 0°C. The reaction mixture was stirred at 0°C for 15 minutes, and then stirred at room temperature for another 15 minutes. After the reaction mixture was cooled to 0°C, a solution of isopropyl((R,S)-(pentafluorophenoxy)-phenoxy-phosphoryl)-L-alaninate (167 mg, 367 μmol) dissolved in dry THF (4 mL) was added dropwise over 10 minutes. The reaction mixture was stirred at 0°C for 30 minutes and at room temperature for 18 hours. The reaction mixture was quenched with saturated NH4Cl aqueous solution (4 mL) and extracted with ethyl acetate (5 mL three times). The combined organic matter was dried, filtered, and concentrated under (Na2SO4). The residue was purified by column chromatography (DCM / MeOH gradient from 100:0 to 90:10) followed by reverse-phase column chromatography (H2O / MeOH gradient from 100:0 to 0:100) to obtain product 7 (a mixture of diastereomers, 35 mg, 58 μmol, 24%) as a white solid. 1 H NMR(300MHz,CD3OD)δ 7.83(s,0.5H), 7.82(s,0.5H), 7.34~7.16(m,5H), 6.15(d,J=18.7Hz,0.5H), 6.13(d,J=18.8Hz,0.5H), 4.99~4.85(overlaps with H2O, m ,1H), 4.65~4.26(m,3H), 4.27~4.12(m,1H), 3.99~3.81(m,1H), 3.42,3.41(2brs,6H), 1.36~1.25(m,3H), 1.24~1.11(m,9H). 19F NMR (282MHz, CD3OD) δ -162.7(s), -163.2(s). 31 PNMR(121MHz,CD3OD)δ 4.08(s), 4.00(s). C 25 H36 FN7O7P[M+H] + The calculated MS(ESI)m / z value was 596.5; the measured value was 596.2. [ka] i)a) N-methylcyclopropylamine hydrochloride, Et3N, MeOH, 100 b) NH4OH, MeOH, 100℃; ii) Isopropyl((R,S)-(pentafluorophenoxy)-phenoxy-phosphoryl)-L-alaninate, tBuMgCl, THF, 0℃.

[0198] Example 3. Preparation of isopropyl((((R,S)-(2R,3R,4R,5R)-5-(2-amino-6-(N-methyl-cyclopropylamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate (9).

[0199] Step 1. Preparation of (2R,3R,4R,5R)-5-(2-amino-6-(N-methylcyclopropylamino)-9H-purine-9-yl)-4-fluoro-2-(hydroxymethyl)-4-methyltetrahydrofuran-3-ol (8). To a solution of compound 3 (600 mg, 1.14 mmol) in MeOH (10 mL), N-methylcyclopropylamine hydrochloride (366 mg, 3.40 mmol) and triethylamine (470 μL, 3.40 mmol) were added. The reaction mixture was heated in a sealed tube at 100 °C for 15 hours and then cooled to room temperature. An aqueous solution containing 30% NH4OH (4 mL) was added, and the reaction mixture was heated in a sealed tube at 100 °C for 2 hours, then cooled and concentrated. The residue was purified by column chromatography (DCM / MeOH gradient from 100:0 to 90:10) to obtain product 8 (351 mg, 0.99 mmol, 87%) as a white solid. 1H NMR(300MHz,CD3OD)δ 8.13(s,1H), 6.15(d,J=18.0Hz,1H), 4.40(dd,J=24.3,9.0Hz,1H), 4.06~4.02(m,2H), 3.89~3.83(m ,1H), 3.32(m,3H), 3.18~3.11(m,1H), 1.16(d,J=22.2Hz,3H), 0.96~0.89(m,2H), 0.74~0.69(m,2H). 19 F NMR(282MHz,CD3OD)δ -163.8. C 15 H 22 FN6O3[M+H] + MS(ESI)m / z calculated value: 353.2; measured value: 353.2.

[0200] Step 2. Preparation of isopropyl((((R,S)-(2R,3R,4R,5R)-5-(2-amino-6-(N-methyl-cyclopropylamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate (9). To a solution of compound 8 (200 mg, 0.57 mmol) in dry THF (15 mL) at 0°C, tert-butylmagnesium chloride (1.0 M in THF, 680 μL, 0.68 mmol) was added dropwise over 10 minutes. The reaction mixture was stirred at 0°C for 15 minutes, and then stirred at room temperature for another 15 minutes. The reaction mixture was cooled to 0°C, and a solution of isopropyl((R,S)-(pentafluorophenoxy)-phenoxy-phosphoryl)-L-alaninate (283 mg, 0.62 mmol) dissolved in dry THF (4 mL) was added dropwise over 10 minutes. The reaction mixture was stirred at 0°C for 30 minutes and at room temperature for 18 hours. The reaction mixture was quenched with saturated NH4Cl aqueous solution (4 mL) and extracted with ethyl acetate (5 mL three times). The combined organic matter was dried over Na2SO4 and concentrated. The residue was subjected to column chromatography (DCM / MeOH gradient from 100:0 to 90:10), followed by reverse-phase column chromatography (H2O / MeOH). The product was purified by a gradient from 100:0 to 0:100 to obtain product 9 (a mixture of two diastereoisomers, 160 mg, 0.26 mmol, 45%) as a white solid. 1 H NMR(300MHz,CD3OD)δ 7.85(m,1H), 7.38-7.16(m,5H), 6.18(d,J=18.6Hz) and 6.16(d,J=18.9Hz,1H), 4.95~4.90(overlaps with H2O, m,1H), 4.58~4.47(m,3H), 4.22~ 4.19(m,1H), 3.95~3.87(m,1H), 3.36~3.34(overlaps with MeOH, m,3H), 3.19~3.12(m,1H), 1.32~1.22(m,12H), 0.96~0.89(m,2H), 0.74~0.69 (m, 2H). 31 PNMR(121MHz,CD3OD)δ 4.11(s), 4.02(s). C 27 H 38 FN7O7P[M+H] + MS(ESI)m / z calculated value: 622.2; measured value: 622.2. [ka] i) 2,6-Dichloropurine, tBuOK, tBuOH / MeCN, 65℃; ii) MeNH2, MeOH, 130℃; iii) Isopropyl ((R,S)-(pentafluorophenoxy)-phenoxy-phosphoryl)-L-alaninate, tBuMgCl, THF, 0℃ to room temperature

[0201] Example 4. Preparation of isopropyl((((R,S)-(2R,3R,4R,5R)-5-(2,6-bis-methylamino-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate (12).

[0202] Step 1. Preparation of (2R,3R,4R,5R)-5-(2,6-dichloro-9H-purine-9-yl)-2-(benzoyloxymethyl)-4-fluoro-4-methyltetrahydrofuran-3-ylbenzoate (10). Compound 2,6-dichloropurine (1.30 g, 6.86 mmol) was suspended in t-BuOH (25 mL) under a nitrogen atmosphere. Potassium tert-butoxide (778 mg, 6.92 mmol) was added in small increments, and the reaction mixture was stirred at room temperature. After 1 hour, a solution of bromofuranoside 2 (1.0 g, 2.29 mmol) dissolved in anhydrous MeCN (20 mL) was added, and the reaction mixture was heated overnight at 65°C and then cooled to room temperature. Saturated NH4Cl aqueous solution was added, and the resulting solution was extracted with RINKAN (3 times). The combined organic matter was dried over Na2SO4 and concentrated. The residue was subjected to column chromatography (PE / RINKAN). The product was purified by a gradient from 100:0 to 0:100 to obtain product 10 (148 mg, 0.27 mmol, 12%) as a viscous solid. 1 H NMR(300MHz,CDCl3)δ 8.31(s,1H), 8.12~8.09(m,2H), 8.02~7.99(m,2H), 7.64~7.39(m,6H), 6.38(d,J=17) .2Hz,1H), 6.02(dd,J=21.2,8.9Hz,1H), 4.90~4.68(m,3H), 1.33(d,J=22.4Hz,3H). 19 F NMR(282MHz, CDCl3)δ -158.0. C 25 H 20 Cl2FN4O5[M+H] + The calculated MS(ESI)m / z value was 546.4; the measured value was 546.3.

[0203] Step 2. Preparation of (2R,3R,4R,5R)-5-(2,6-bis-methylamino-9H-purine-9-yl)-4-fluoro-2-(hydroxymethyl)-4-methyltetrahydrofuran-3-ol (11). Compound 10 (148 mg, 0.27 mmol) contains methylamine (33% in EtOH, 3 The 0 mL solution was heated in a sealed tube at 130°C for 4 days, then cooled to room temperature and concentrated. The residue was purified by column chromatography (DCM / MeOH gradient from 100:0 to 50:50) followed by reverse-phase column chromatography (H2O / MeOH gradient from 100:0 to 0:100) to obtain Product 11 (33 mg, 0.10 mmol, 37%) as a white solid. 1 H NMR(300MHz,CD3OD)δ 8.00(s,1H), 6.12(d,J=18.5Hz,1H), 4.51(dd,J=24.4,9.5Hz,1H), 4.06~3.85(m,3H), 3.04(s,3H), 2.93(s,3H), 1.20(d,J=22.4Hz,3H). 19 F NMR(282MHz,CD3OD)δ -163.2. C 13 H 20 FN6O3[M+H] + The calculated MS(ESI)m / z value is 327.2; the measured value is 327.2.

[0204] Step 3. Preparation of isopropyl((((R,S)-(2R,3R,4R,5R)-5-(2,6-bis-methylamino-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate (12). To a solution of compound 11 (55 mg, 0.17 mmol) at 0°C in dry THF (2 mL), tert-butyl magnesium chloride (1 M in THF, 304 L, 0.30 mmol) was added dropwise over 10 minutes. The reaction mixture was stirred at 0°C for 15 minutes, and then stirred at room temperature for another 15 minutes. The solution was cooled to 0°C, and a solution of isopropyl ((R,S)-(pentafluorophenoxy)-phenoxy-phosphoryl)-L-alaninate (115 mg, 0.25 mmol) dissolved in dry THF (1 mL) was added dropwise over 10 minutes. The mixture was slowly warmed to room temperature and stirred for 4 days. The reaction product was quenched with saturated NH4Cl aqueous solution and extracted with ethyl acetate (3 times). The combined organic matter was dried over Na2SO4 and concentrated. The residue was purified by column chromatography (DCM / MeOH gradient from 100:0 to 50:50) to obtain product 12 (a mixture of diastereomers, 13 mg, 0.02 mmol, 13%) as a white solid. 1 H NMR(300MHz,CD3OD)δ 7.78(s,1H), 7.35~7.12(m,5H), 6.13(d,J=19.1Hz,0.53H), 6.10(d,J=19.2Hz,0.47H), 4.99~4.78(overlaps with H2O, m,1 H), 4.72~4.46(m,3H), 4.24~4.15(m,1H), 3.79~3.92(m,1H), 3.02(brs,3H), 2.92(s+s,3H), 1.29~1.11(m,12H). 19 F NMR (282MHz, CD3OD) δ -162.0(s), -162.3(s). 31 PNMR(121MHz,CD3OD)δ 3.97(s), 3.89(s). C 25 H 36 FN7O7P[M+H] + The calculated MS(ESI)m / z value was 596.6; the measured value was 596.2. [ka] i) TIPDSCl2, imidazole, DMF; ii) Isobutyryl chloride, pyridine; iii) TBAF, THF; iv) Isopropyl ((R,S)-(pentafluorophenoxy)-phenoxy-phosphoryl)-L-alaninate, tBuMgCl, THF, 0℃.

[0205] Example 5. Preparation of isopropyl((((R,S)-(2R,3R,4R,5R)-5-(2-isobutylamide-6-methylamino-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate (16).

[0206] Step 1. Preparation of compound 13. 1,3-Dichloro-1,1,3,3-tetraisopropyldisiloxane (300 μL, 0.94 mmol) was added to a 6 mL dry DMF solution of compound 4 (286 mg, 0.92 mmol) and imidazole (370 mg, 5.43 mmol) at 0°C. The reaction mixture was stirred at room temperature for 2 hours, diluted with  (50 mL), and the suspension was washed with saturated NH4Cl aqueous solution and brine (40 mL each). The organic matter was dried over Na2SO4 and concentrated. The residue was purified by column chromatography (PE /  7:3 to 3:7 gradient) to obtain product 13 (283 mg, 0.51 mmol, 56%) as a white solid. 24 H 44 FN6O4Si2[M+H] + The calculated MS(ESI)m / z value was 555.8; the measured value was 555.2.

[0207] Step 2. Preparation of compound 14. Isobutyryl chloride (38 μL, 0.36 mmol) was added to a solution of compound 13 (200 mg, 0.36 mmol) at 0°C in dried pyridine (3 mL). The reaction mixture was stirred at room temperature for 2 hours. The reaction product was quenched by adding water (500 μL). The mixture was concentrated and evaporated simultaneously with toluene (10 mL three times). The residue was purified by column chromatography (PE / Â1 1:0 to 1:1 gradient) to obtain product 14 (99 mg, 0.16 mmol, 44%) as a white solid. 28 H 50 FN6O5Si2[M+H] + The calculated MS(ESI)m / z value was 625.9; the measured value was 625.3.

[0208] Step 3. (2R,3R,4R,5R)-5-(2-isobutylamide-6-methylamino Preparation of -9H-purine-9-yl)-4-fluoro-2-(hydroxymethyl)-4-methyltetrahydrofuran-3-ol (15). To a solution of compound 14 (90 mg, 0.14 mmol) in dried THF (2 mL), tetrabutylammonium fluoride (1 M in THF, 38 μL, 0.38 mmol) was added. The mixture was stirred at room temperature for 2 hours and then concentrated. The residue was purified by column chromatography (DCM / MeOH gradient from 10:0 to 9:1) followed by reverse-phase column chromatography (H2O / MeOH gradient from 100:0 to 0:100) to obtain product 15 (42 mg, 0.11 mmol, 77%) as a white solid. 1 H NMR(300MHz,CD3OD)δ 8.31(s,1H), 6.29(d,J=17.9Hz,1H), 4.70~4.60(m,1H), 4.07~3.98(m,2H), 3. 89(dd,J=12.5,3.4Hz,1H), 3.10(brs,3H), 2.87(brs,1H), 1.23~1.16(m,9H). 19 F NMR(282MHz,CD3OD)δ -163.8. C 16 H 24 FN6O4[M+H] + The calculated MS(ESI)m / z value was 383.4; the measured value was 383.2.

[0209] Step 4. Preparation of isopropyl((((R,S)-(2R,3R,4R,5R)-5-(2-isobutylamide-6-methylamino-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate (16). To a solution of compound 15 (27 mg, 0.07 mmol) in dry THF (1 mL) at 0°C, t-butylmagnesium chloride (1.0 M in THF, 130 μL, 0.13 mmol) was added dropwise over 10 minutes. The reaction mixture was stirred at 0°C for 15 minutes, and then stirred at room temperature for another 15 minutes. The reaction mixture was cooled to 0°C, and a solution of isopropyl((R,S)-(pentafluorophenoxy)-phenoxy-phosphoryl)-L-alaninate (50 mg, 0.11 mmol) dissolved in dry THF (1 mL) was added dropwise over 10 minutes. The reaction mixture was stirred at 0°C for 30 minutes, then stirred at room temperature for 18 hours, and then quenched with saturated NH4Cl aqueous solution (2 mL) and extracted with ethyl acetate (5 mL three times). The combined organic matter was dried over Na2SO4 and concentrated. The residue was purified by column chromatography (DCM / MeOH gradient from 100:0 to 95:5) followed by reverse-phase column chromatography (H2O / MeOH gradient from 100:0 to 0:100) to obtain product 16 (a mixture of two diastereoisomers, 25 mg, 0.04 mmol, 54%) as a white solid. 1 H NMR(300MHz,CD3OD)δ 8.05(s,1H), 7.33~7.13(m,5H), 6.27(d,J=18.6Hz) and 6.21(d,J=19.1Hz,1H), 5.10~4.95(m,1H), 4.93~4.78(overlapping with H2O, m ,1H), 4.60~4.42(m,2H), 4.26~4.18(m,1H), 3.90~3.80(m,1H), 3.09(brs,3H), 2.84~2.80(m,1H), 1.33~1.15(m,18H). 31 PNMR(121MHz,CD3OD)δ 3.69(s). 31 PNMR(121MHz,CD3OD)δ 4.11(s), 3.99(s). C 28 H40 FN7O8P[M+H] + The calculated MS(ESI)m / z value was 652.6; the measured value was 652.3. [ka] i) N-methylethylamine, MeOH, 100℃; ii) Isopropyl ((R,S)-(pentafluorophenoxy)-phenoxy-phosphoryl)-L-alaninate, tBuMgCl, THF, 0℃.

[0210] Example 6. Preparation of isopropyl((((R,S)-(2R,3R,4R,5R)-5-(2-amino-6-(N-methyl-ethylamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate (18).

[0211] Step 1. Preparation of (2R,3R,4R,5R)-5-(2-amino-6-(N-methyl-ethylamino)-9H-purine-9-yl)-4-fluoro-2-(hydroxymethyl)-4-methyltetrahydrofuran-3-ol (17). N-methylethylamine (245 μL, 2.90 mmol) was added to a solution of compound 3 (150 mg, 0.29 mmol) in MeOH (4 mL). The reaction mixture was heated at 100 °C in a sealed tube for 15 hours, then cooled to room temperature and concentrated. The residue was purified by column chromatography (DCM / MeOH gradient from 100:0 to 90:10) to obtain product 31 (89 mg, 0.26 mmol, 89%) as a white solid. 1 H NMR(300MHz,CD3OD)δ 8.06(s,1H), 6.13(d,J=18.0Hz,1H), 4.40(dd,J=24.9,8.7Hz,1H), 4.11~4.01(m,4H), 3.98~3.83(m,1H), 3.34(br.s,3H), 1.24~1.11(m,6H). 19 F NMR(282MHz,CD3OD)δ -163.7. C 14 H 22 FN6O3[M+H]+ The calculated MS(ESI)m / z value is 341.2; the measured value is 341.2.

[0212] Step 2. Preparation of isopropyl((((R,S)-(2R,3R,4R,5R)-5-(2-amino-6-(N-methyl-ethylamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate (18). To a solution of compound 17 (30 mg, 0.09 mmol) in dry THF (2 mL) at 0°C, tert-butylmagnesium chloride (1.0 M in THF, 110 μL, 0.11 mmol) was added dropwise over 10 minutes. The reaction mixture was stirred at 0°C for 15 minutes, and then stirred at room temperature for another 15 minutes. The reaction mixture was cooled to 0°C, and a solution of isopropyl((R,S)-(pentafluorophenoxy)-phenoxy-phosphoryl)-L-alaninate (48 mg, 0.11 mmol) dissolved in dry THF (1 mL) was added dropwise over 10 minutes. The reaction mixture was stirred at 0°C for 30 minutes and at room temperature for 18 hours. The reaction mixture was quenched with saturated NH4Cl aqueous solution (4 mL) and extracted with ethyl acetate (5 mL three times). The combined organic matter was dissolved in Na2SO4. The solution was dried and concentrated in 4°C. The residue was purified by column chromatography (DCM / MeOH gradient from 100:0 to 90:10) to obtain product 18 (a mixture of two diastereoisomers, 22 mg, 0.04 mmol, 40%) as a white solid. 1 H NMR(300MHz,CD3OD)δ 7.69(m,1H), 7.26~7.04(m,5H), 6.05(d,J=18.6Hz) and 6.03(d,J=18.9Hz,1H), 4.86~4.79(overlapping with H2O, m ,1H), 4.50~4.32(m,3H), 4.12~4.06(m,1H), 3.96~3.79(m,3H), 3.25(br.s,3H), 1.24~1.02(m,15H). 31 PNMR(121MHz,CD3OD)δ 4.07(s), 4.00(s). C 26 H 38 FN7O7P[M+H] +The calculated MS(ESI)m / z value was 609.3; the measured value was 609.2. [ka] i) N-methylpropylamine, MeOH, 100℃; ii) Isopropyl((R,S)-(pentafluorophenoxy)-phenoxy-phosphoryl)-L-alaninate, tBuMgCl, THF, 0℃.

[0213] Example 7. Preparation of isopropyl((((R,S)-(2R,3R,4R,5R)-5-(2-amino-6-(N-methyl-propylamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate (20).

[0214] Step 1. Preparation of (2R,3R,4R,5R)-5-(2-amino-6-(N-methyl-propylamino)-9H-purine-9-yl)-4-fluoro-2-(hydroxymethyl)-4-methyltetrahydrofuran-3-ol (19). N-methylpropylamine (295 μL, 2.90 mmol) was added to a solution of compound 3 (150 mg, 0.29 mmol) in MeOH (4 mL). The reaction mixture was heated in a sealed tube at 100 °C for 15 hours, then cooled to room temperature and concentrated. The residue was purified by column chromatography (DCM / MeOH gradient from 100:0 to 90:10) followed by reverse-phase column chromatography (H2O / MeOH gradient from 100:0 to 0:100) to obtain product 19 (80 mg, 0.23 mmol, 78%) as a white solid. 1 H NMR(300MHz,CD3OD)δ 8.04(s,1H), 6.13(d,J=18.3,1H), 4.40(dd,J=24.2,9.2Hz,1H), m,4.06~3.84( m,5H), 1.68(sept,J=7.5Hz,2H), 1.15(d,J=22.2Hz,3H), 0.93(t,J=7.5Hz,3H). 19 F NMR(282MHz,CD3OD)δ -163.8. C 15H 24 FN6O3[M+H] + The calculated MS(ESI)m / z value is 355.2; the measured value is 355.2.

[0215] Step 2. Preparation of isopropyl((((R,S)-(2R,3R,4R,5R)-5-(2-amino-6-(N-methyl-propylamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate (20). To a solution of compound 19 (30 mg, 0.09 mmol) in dry THF (2 mL) at 0°C, tert-butylmagnesium chloride (1.0 M in THF, 110 μL, 0.11 mmol) was added dropwise over 10 minutes. The reaction mixture was stirred at 0°C for 15 minutes, and then stirred at room temperature for another 15 minutes. The reaction mixture was cooled to 0°C, and a solution of isopropyl((R,S)-(pentafluorophenoxy)-phenoxy-phosphoryl)-L-alaninate (46 mg, 0.11 mmol) dissolved in dry THF (1 mL) was added dropwise over 10 minutes. The reaction mixture was stirred at 0°C for 30 minutes and at room temperature for 18 hours. The reaction mixture was quenched with saturated NH4Cl aqueous solution (4 mL) and extracted with ethyl acetate (5 mL three times). The combined organic matter was dried over Na2SO4 and concentrated. The residue was purified by column chromatography (DCM / MeOH gradient from 100:0 to 90:10) to obtain product 20 (a mixture of two diastereoisomers, 22 mg, 0.03 mmol, 33%) as a white solid. 1 H NMR(300MHz,CD3OD)δ 7.78,7.77(s+s,1H), 7.37~7.13(m,5H), 6.15(d,J=18.6Hz) and 6.13(d,J=18.9Hz,1H), 4.97~4.89(overlaps with H2O, m,1H), 4.63~4.30(m,3H), 4. 22~4.14(m,1H), 4.02~3.84(m,2H), 1.74~1.63(3H,m), 1.32~1.27(m,3H), 1.23~1.13(m,9H), 0.94(t,J=7.4Hz) and 0.93(t,J=7.4Hz,3H). 31PNMR(121MHz,CD3OD)δ 4.05(s), 4.00(s). C 27 H 40 FN7O7P[M+H] + The calculated MS(ESI)m / z value was 623.3; the measured value was 623.2. [ka] i) a) N-methylcyclobutylamine hydrochloride, Et3N, MeOH, 100℃; b) NH4OH, MeOH, 100℃; ii) Isopropyl ((R,S)-(pentafluorophenoxy)-phenoxy-phosphoryl)-L-alaninate, tBuMgCl, THF, 0℃.

[0216] Example 8. Preparation of isopropyl((((R,S)-(2R,3R,4R,5R)-5-(2-amino-6-(N-methyl-cyclobutylamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate (22).

[0217] Step 1. Preparation of (2R,3R,4R,5R)-5-(2-amino-6-(N-methylcyclobutylamino)-9H-purine-9-yl)-4-fluoro-2-(hydroxymethyl)-4-methyltetrahydrofuran-3-ol (21). To a solution of compound 3 (150 mg, 0.29 mmol) in MeOH (4 mL), N-methylcyclobutylamine hydrochloride (105 mg, 0.90 mmol) and triethylamine (190 μL, 1.00 mmol) were added. The reaction mixture was heated in a sealed tube at 100 °C for 15 hours and then cooled to room temperature. An aqueous solution containing 30% NH4OH (1 mL) was added, and the reaction mixture was heated in a sealed tube at 100 °C for 2 hours, then cooled and concentrated. The residue was purified by column chromatography (DCM / MeOH gradient from 100:0 to 90:10) to obtain product 21 (90 mg, 0.25 mmol, 86%) as a pale yellow solid. 1H NMR(300MHz,CD3OD)δ 8.09(s,1H), 6.14(d,J=18.0Hz,1H), 5.80~5.70(m,1H), 4.44~4.33(m,1H), 4.06~4.02(m,2H), 3.88~3.84(m,1H), 3.34(s,3H), 2.38~2.19(m,4H), 1.79~1.71(m,2H), 1.17(d,J=22.2Hz,3H). 19 F NMR(282MHz,CD3OD)δ -163.8. C 16 H 24 FN6O3[M+H] + MS(ESI)m / z calculated value: 367.2; measured value: 367.2.

[0218] Step 2. Preparation of isopropyl((((R,S)-(2R,3R,4R,5R)-5-(2-amino-6-(N-methyl-cyclobutylamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate (22). To a solution of compound 21 (50 mg, 0.14 mmol) in dry THF (2 mL) at 0°C, tert-butylmagnesium chloride (1.0 M in THF, 210 μL, 0.21 mmol) was added dropwise over 10 minutes. The reaction mixture was stirred at 0°C for 15 minutes, and then stirred at room temperature for another 15 minutes. The reaction mixture was cooled to 0°C, and a solution of isopropyl((R,S)-(pentafluorophenoxy)-phenoxy-phosphoryl)-L-alaninate (74 mg, 0.16 mmol) dissolved in dry THF (2 mL) was added dropwise over 10 minutes. The reaction mixture was stirred at 0°C for 30 minutes and at room temperature for 18 hours. The reaction mixture was quenched with saturated NH4Cl aqueous solution (4 mL) and extracted with ethyl acetate (5 mL three times). The combined organic matter was dried over Na2SO4 and concentrated. The residue was purified by column chromatography (DCM / MeOH gradient from 100:0 to 90:10) followed by reverse-phase column chromatography (H2O / MeOH gradient from 100:0 to 0:100) to obtain product 22 (a mixture of two diastereoisomers, 24 mg, 0.04 mmol, 28%) as a white solid. 1H NMR(300MHz,CD3OD)δ 7.79(s,0.2H), 7.77(s,0.8H), 7.38~7.12(m,5H), 6.18(d,J=17.6Hz) and 6.16(d,J=17.5Hz,1H), 4.95~4.81(m,2H), 4.62~4.43( m,3H), 4.25~4.18(m,1H), 3.96~3.83(m,1H), 3.38(s) and 3.36(s,3H), 2.38~2.21(m,4H), 1.75~1.63(m,2H), 1.32~1.16(m,12H). 31 PNMR(121MHz,CD3OD)δ 4.04(s), 3.97(s). C 28 H 40 FN7O7P[M+H] + The calculated MS(ESI)m / z value was 636.3; the measured value was 636.2.

[0219] Modification of the 2-amino moiety in the active compound Those skilled in the art can add substituents to the 2-aminopurine moiety by methods well known to those skilled in the art. One non-limiting process is provided hereby, and others can be readily adapted. ((2R,3R,4R,5R)-3-(benzoyloxy)-5-bromo-4-fluoro-4-methyltetrahydrofuran-2-yl)methylbenzoate by a mixture of commercially available 2,6-dichloropurine, a base and an organic solvent. The compound (2R,3R,4R,5R)-5-(2,6-dichloro-9H-purine-9-yl)-2-(benzoyloxymethyl)-4-fluoro-4-methyltetrahydrofuran-3-ylbenzoate is produced by treatment at high temperature. In one embodiment, the base is potassium tert-butoxide. In one embodiment, the organic solvent mixture includes tert-butanol and acetonitrile. The compound (2R,3R,4R,5R)-5-(2,6-dichloro-9H-purine-9-yl)-2-(benzoyloxymethyl)-4-fluoro-4-methyltetrahydrofuran-3-ylbenzoate is treated with an amine, a base and an organic solvent at ambient temperature to produce 2-chloro-N 6-A substituted purine is produced. In one embodiment, the amine is methylamine. In one embodiment, the base is triethylamine. In one embodiment, the organic solvent is ethanol. Those skilled in the art will recognize that treatment with the amine and base simultaneously removes the benzoate group of the nucleoside, producing a deprotected furanose moiety. Subsequently, in a sealed tube, at a high temperature of about 100°C, 2-chloro-N 6 - Substituted purines are treated with amines and organic solvents, and the N of the present invention 2 ,N 6 - Disubstituted purine nucleosides can be produced. In one embodiment, the amine is methylamine. In one embodiment, the organic solvent is ethanol. 2 ,N 6 - Disubstituted purine nucleosides can be treated at low temperatures with a base, isopropyl((R,S)-(pentafluorophenoxy)-phenoxy-phosphoryl)-L-alaninate, and an organic solvent to produce compounds of formulas I to V. In one embodiment, the base is tert-butyl magnesium chloride. In one embodiment, the organic solvent is tetrahydrofuran.

[0220] Preparation of stereospecific phosphoenantiomers Certain active compounds described herein have a chiral phosphorus moiety. Any of the active compounds described herein can be provided in isolated phosphorus enantiomer form, for example, as an enantiomer of at least 80%, 90%, 95%, or 98% R or S, using methods known to those skilled in the art. There are many publications describing methods for obtaining such compounds, including, but are not limited to, column chromatography, as described in Example 17 below and, for example, in U.S. Patents 8,859,756, 8,642,756, and 8,333,309 to Ross, et al.

[0221] Example 9. Separation of stereoisomers of compound 5. The stereoisomers of compound 5 were separated on a Phenominex Luna column using the following conditions. Column: Phenominex Luna 5 micron C18(2) 250×10mm Part number OOG-4252-BO Sample concentration: Approximately 50 mg / ml in acetonitrile Injection volume: 50μl Mobile phase A: HPLC-grade water Mobile phase B: HPLC-grade acetonitrile Flow rate: 5ml / min UV: 283nm gradient:

[0222] [Table 1] Execution time: 45 minutes Column temperature: 40℃

[0223] Figure 1 shows the chromatogram of half of the sample.

[0224] The combined fractions were evaluated using an analytical column under the following conditions. Column: Phenominex Luna 5 micron C18(2) 250×2mm Part number OOG-4252-BO Injection volume: 10μl Mobile phase A: HPLC-grade water Mobile phase B: HPLC-grade acetonitrile Flow rate: 0.2ml / min UV: 283nm gradient:

[0225] [Table 2] Execution time: 45 minutes Column temperature: 40℃

[0226] The combined fractions for each stereoisomer were evaporated to dryness using a rotary evaporator (rotovap) at a bath temperature of 30°C. The resulting solid was dissolved in 1 ml of acetonitrile, and 1 The solution was transferred to a 0.7 ml microcentrifuge tube, and the solvent was evaporated by vacuum centrifugation at 30°C.

[0227] The data for the final sample is as follows: 1. First elution peak: Compound 5, No. 1 (5-1) (21.7 mg - 97.8% ee). 2. Second elution peak: Compound 5, No. 2 (5-2) (13.2 mg - 95.9% ee).

[0228] The final weights of the first and second peaks are equal to their percentages in the original mixture. This corresponds to 62.2% and 37.8% respectively.

[0229] Stereospecific synthesis of compounds of formulas I to VII [ka]

[0230] Example 10. Preparation of (2R,3R,4R,5R)-5-(2-amino-6-chloro-9H-purine-9-yl)-2-(hydroxymethyl)-4-fluoro-4-methyltetrahydrofuran-3-ol (23).

[0231] Step 1. Preparation of (2R,3R,4R,5R)-5-(2-amino-6-chloro-9H-purine-9-yl)-2-(hydroxymethyl)-4-fluoro-4-methyltetrahydrofuran-3-ol (23). Compound (2R,3R,4R,5R)-5-(2-amino-6-chloro-9H-purine-9-yl)-2-(benzoyloxymethyl)-4-fluoro-4-methyltetrahydrofuran-3-ylbenzoate, 3 (80 g, 140 mmol) was added to a methanol (7 M, 800 mL) solution of trimethylamine and stirred overnight at room temperature. After concentrating the mixture, it was purified by column chromatography (DCM:MeOH=100:1) to obtain (2R,3R,4R,5R)-5-(2-amino-6-chloro-9H-purine-9-yl)-2-(hydroxymethyl)-4-fluoro-4-methyltetrahydrofuran-3-ol (23) (40 g, 90%). [ka]

[0232] Example 11. Preparation of ((((S)-(2R,3R,4R,5R)-5-(2-amino-6-(methylamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate. [ka]

[0233] Step 1. Preparation of (2R,3R,4R,5R)-5-(2-amino-6-(methylamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-3-ol (4). (2R,3R,4R,5R)-5-(2-amino-6-chloro-9H-purine-9-yl)-2-(hydroxymethyl)-4-fluoro-4-methyltetrahydrofuran-3-ol (2.0 g, 1.0 equivalent) was dissolved in dioxane (15 mL) and then 5.0 equivalents of MeNH2 aqueous solution were added. After stirring overnight at room temperature, TLC showed that the starting material had been consumed. The mixture was concentrated and purified by column chromatography (DCM:MeOH = 40:1 to 30:1) to obtain (2R,3R,4R,5R)-5-(2-amino-6-(methylamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-3-ol (1.6 g, 81.6%) as a white powder. [M+H] + =313.5 [ka]

[0234] Step 2. Preparation of ((((S)-(2R,3R,4R,5R)-5-(2-amino-6-(methylamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate. The compounds (2R,3R,4R,5R)-5-(2-amino-6-(methylamino)-9H-purin-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-3-ol (1.47 g, 1.0 equivalent) and PPAL-S (2.35 g, 1.1 equivalent) were dissolved in anhydrous THF (29 mL). After cooling the mixture to -10°C, t-BuMgCl (5.8 mL, 1.7 M, 2.1 equivalents) was slowly added under an N2 blanket. After stirring at room temperature for 45 minutes, the mixture was quenched with saturated NH4Cl water and extracted with ELISA (20 mL three times). The combined organic layer was washed with water and brine (30 mL), dried over anhydrous Na2SO4, and concentrated. The crude product was purified by column chromatography (DCM:MeOH = 50:1 to 20:1) to obtain ((((S)-(2R,3R,4R,5R)-5-(2-amino-6-(methylamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate (1.1 g, 40.3%) as a white powder. 1 H NMR(400MHz,CD3OD)δ 7.81(s,1H), 7.33~7.16(m,5H), 6.10(d,J=18.4Hz,1H), 4.90~4.84(m ,5H), 4.55~4.46(m,3H), 4.20~4.16(m,1H), 3.91~3.87(m,1H), 3.30(m,1H), 3.03(s,3H), 1.30~1.20(m,12H). [M+H] + = 582.8. [ka]

[0235] Example 12. Preparation of isopropyl((((S)-(2R,3R,4R,5R)-5-(2-amino-6-(dimethylamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate (25). [ka]

[0236] Step 1. Preparation of (2R,3R,4R,5R)-5-(2-amino-6-(dimethylamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-3-ol. (2R,3R,4R,5R)-5-(2-amino-6-chloro-9H-purine-9-yl)-2-(hydroxymethyl)-4-fluoro-4-methyltetrahydrofuran-3-ol (2.8 g, 8 mmol) was dissolved in dioxane (20 mL) and dimethylamine aqueous solution (5 mL) was added. After stirring at room temperature for 3 hours, TLC showed that the starting material had been consumed. The mixture was concentrated and purified by column chromatography (DCM:MeOH=60:1) to obtain (2R,3R,4R,5R)-5-(2-amino-6-(dimethylamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-3-ol (2.2 g). 1 H NMR(400MHz,CD3OD)δ 8.08(s,1H), 6.13(d,J=18.0Hz,1H), 4.43(dd,J=9.2,9.2Hz,1H), 4.06(d,J =10.8Hz,2H), 3.90(m,1H), 3.37(s,3H), 3.06(s,3H), 1.18(d,J=22Hz,3H). [ka]

[0237] Step 2. Preparation of isopropyl((((S)-(2R,3R,4R,5R)-5-(2-amino-6-(dimethylamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate (25). The compounds (2R,3R,4R,5R)-5-(2-amino-6-(dimethylamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-3-ol (8 g, 1.0 equivalent) and PPAL-S (11.1 g, 1 equivalent) were dissolved in anhydrous THF (100 mL). The mixture was cooled to -5°C to 0°C, and t-BuMgCl (30.5 mL, 1.7 M, 2.1 equivalents) was slowly added under an N2 atmosphere. After stirring at room temperature for 2 hours, the mixture was quenched with saturated NH4Cl aqueous solution and extracted with ethyl acetate (3 times with 70 mL). The combined organic layer was washed with water and brine (30 mL), dried over anhydrous Na2SO4, and concentrated. The crude product was purified by column chromatography (DCM:MeOH=50:1) to obtain isopropyl((((S)-(2R,3R,4R,5R)-5-(2-amino-6-(dimethylamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate (9.5g, 65%) as a white powder. 1 H NMR(400MHz,CD3OD)δ 7.81(s,1H), 7.35~7.19(m,5H), 6.15(d,J=18.8Hz,1H), 4.90(m,1H), 4.54~4.49(m,3H) , 4.22~4.19(m,1H), 3.90(m,1H), 3.43(s,3H), 1.32(d,J=7.2Hz,3H), 1.24~1.17(m,9H). 31 P NMR (160 MHz, CD3OD) δ 3.89. [ka]

[0238] Example 13. Preparation of isopropyl((((R)-(2R,3R,4R,5R)-5-(2-amino-6-(dimethylamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate (26).

[0239] The compounds (2R,3R,4R,5R)-5-(2-amino-6-(dimethylamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-3-ol (3g, 1.0 equivalent) and PPAL-R (4.17g, 1 equivalent) were dissolved in anhydrous THF (60mL). The mixture was cooled to -5°C to 0°C, and t-BuMgCl (11.4mL, 1.7M, 2.1 equivalents) was slowly added under an N2 atmosphere. After stirring at room temperature for 16 hours, the mixture was quenched with saturated NH4Cl aqueous solution and ethyl alcohol (50mL) was added. Extraction was performed three times. The combined organic layers were washed with water and brine (30 mL), dried over anhydrous Na2SO4, and concentrated. The crude product was purified by column chromatography (DCM:MeOH=50:1) to obtain isopropyl((((R)-(2R,3R,4R,5R)-5-(2-amino-6-(dimethylamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate (2.2 g, 41%) as a white powder. 1 H NMR(400MHz,CD3OD)δ 7.8(s,1H), 7.35~7.29(m,5H), 6.18(d,J=18.8Hz,1H), 4.92(m,1H), 4.60(m,1H), 4. 51~4.23(m,3H), 3.90(m,1H), 3.44(s,6H), 1.29(d,J=6Hz,3H), 1.22~1.16(m,10H). 31 P NMR (160 MHz, CD3OD) δ 3.98. [ka]

[0240] Example 14. Preparation of isopropyl((((S)-(2R,3R,4R,5R)-5-(2-amino-6-(methylcyclopropaneamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate. [ka]

[0241] Step 1: Preparation of (2R,3R,4R,5R)-5-(2-amino-6-(methylcyclopropaneamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-3-ol (8). K2CO3 (53 g, 500 mmol) was added to an aqueous solution of N-methylcyclopropanamine hydrochloride (100 mL). After stirring at room temperature for 10 minutes, a solution of (2R,3R,4R,5R)-5-(2-amino-6-chloro-9H-purine-9-yl)-2-(hydroxymethyl)-4-fluoro-4-methyltetrahydrofuran-3-ol (35 g, 109 mmol) in dioxane (300 mL) was added. The mixture was stirred at room temperature for 16 hours. After stirring, HPLC indicated that the above reaction was complete. The mixture was concentrated and purified by column chromatography (DCM:MeOH=60:1) to obtain (2R,3R,4R,5R)-5-(2-amino-6-(methylcyclopropaneamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-3-ol (30g, 82%). 1 H NMR(400MHz,CD3OD)δ 8.16(s,1H), 6.17(d,J=18.0Hz,1H), 4.41(dd,J=9.2,9.2Hz,1H), 4.06(m,2H), 3.90 (m,1H), 3.37(s,3H), 3.16(m,1H), 1.18(d,J=22.4Hz,3H), 0.94(m,2H), 0.74(m,2H). [M+H] + = 353.2. [ka]

[0242] Step 2: Preparation of isopropyl((((S)-(2R,3R,4R,5R)-5-(2-amino-6-(methylcyclopropaneamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate. The compounds (2R,3R,4R,5R)-5-(2-amino-6-(methylcyclopropaneamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-3-ol (8 g, 1.0 equivalent) and PPAL-S (10.3 g, 1 equivalent) were dissolved in anhydrous THF (100 mL). After cooling the mixture to -5°C to 0°C, t-BuMgCl (28 mL, 1.7 M, 2.1 equivalents) was slowly added under an N2 atmosphere. The mixture was stirred at room temperature for 1 hour, quenched with saturated NH4Cl aqueous solution, and extracted with ELISA (3 times with 70 mL). The combined organic layers were washed with water and brine (30 mL), dried over anhydrous Na2SO4, and concentrated. The crude product was purified by column chromatography (DCM:MeOH = 100:1 to 50:1) to obtain isopropyl((((S)-(2R,3R,4R,5R)-5-(2-amino-6-(methylcyclopropaneamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate (9.5 g, 65%) as a white powder. 1 H NMR(400MHz,CD3OD)δ 7.86(s,1H), 7.35~7.19(m,5H), 6.17(d,J=19.2Hz,1H), 4.91(m,1H), 4.52(m,3H), 4.21(m,1H), 3 .93(m,1H), 3.35(s,3H), 3.16(m,1H), 2.0(s,1H), 1.26~1.16(m,12H), 0.93(m,2H), 0.73(m,2H). 31 P NMR (160 MHz, CD3OD) δ 3.90 [ka]

[0243] Example 15. Preparation of isopropyl((((R)-(2R,3R,4R,5R)-5-(2-amino-6-(methylcyclopropaneamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate.

[0244] The compounds (2R,3R,4R,5R)-5-(2-amino-6-(methylcyclopropaneamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-3-ol (3g, 1.0 equivalent) and PPAL-R (2.8g, 1 equivalent) were dissolved in anhydrous THF (60mL). After cooling the mixture to -5°C to 0°C, t-BuMgCl (7.6mL, 1.7M, 2.1 equivalents) was slowly added under N2. The mixture was then stirred at room temperature for 1 hour, quenched with saturated NH4Cl aqueous solution, and extracted with ethyl acetate (3 times with 50mL). The combined organic layers were washed with water and brine (30mL), dried over anhydrous Na2SO4, and concentrated. The crude product was purified by column chromatography (DCM:MeOH = 100:1 to 50:1) to obtain the product (3g, 55%) as a white powder. 1 H NMR(400MHz,CD3OD)δ 7.81(s,1H), 7.30~7.25(m,5H), 6.16(d,J=24.8Hz,1H), 4.84(m,1H), 4.84~4.50(m,3H), 4.22~4.19(m, 1H), 3.88(m,1H), 3.33(s,3H), 3.14(m,1H), 2.0(s,1H), 1.28~1.13(m,12H), 0.92(m,2H), 0.90(m,2H). 31 P NMR (160 MHz, CD3OD) δ 3.99. [ka]

[0245] Example 16. Preparation of compound 32. [ka]

[0246] Step 1. Preparation of compound 29. To a 30 mL solution of pyridine (6 g, 1.0 equivalent), TIPDSCl2 (4.35 g, 1.5 equivalents) was added at 0°C. After stirring at room temperature for 4 hours, TLC showed that the starting material had been consumed. The mixture was diluted with siRNA, washed with 1 M aqueous HCl, saturated aqueous NaHCO3, and brine, dried over anhydrous Na2SO4, and concentrated to obtain 29 g (6.3 g, 100%) as a yellow oil. [ka]

[0247] Step 2. Preparation of compound 30. Isobutyryl chloride (209 mg, 1.5 equivalents) was added at 0°C to a mixture of compound 29 (800 mg, 1.0 equivalent), DMAP (16 mg, 0.1 equivalent), pyridine (1.6 mL), and DCM (10 mL). After stirring at room temperature for 2 hours, TLC showed that the starting materials had been consumed. The mixture was quenched with water, washed with 1 M aqueous HCl, saturated aqueous NaHCO3, and brine, dried over anhydrous Na2SO4, and concentrated. The crude product was purified by column chromatography to obtain product 30 (563 mg, 62.3%) as a white oil. 1 H NMR (400MHz, CDCl3) δ 7.98(s,1H), 787(s,1H), 6.20(d,J=16.0Hz,1H), 4.32~4.07(m,4H), 3.50(s,6H), 2.3(m,1H), 1.29~1.05(m,45H). [ka]

[0248] Step 3. Preparation of compound 31. Et3N·3HF (706 mg, 5 equivalents) and Et3N (890 mg, 10 equivalents) were added to a mixture of 30 (560 mg, 1.0 equivalent) in THF (10 mL) at room temperature. After stirring at room temperature for 1.5 hours, TLC showed that the starting material had been consumed. The mixture was concentrated and purified by column chromatography to obtain 31 (288 mg, 83 equivalents) as a white powder. %) was obtained. 1 H NMR (400MHz, CDCl3) δ 7.72(s,1H), 5.96(d,J=44.0Hz,1H), 5.22(m,1H), 4.13~3.99(m,4H), 3.42(s,6H), 2.83~2.63(m,2H), 1.29~1.17(m,9H). [ka]

[0249] Step 4. Preparation of compound 32. Compound 31 (280 mg, 1.0 equivalent) and PPAL-S (320 mg, 1 equivalent) were dissolved in anhydrous THF (10 mL). After cooling the mixture to -5°C, t-BuMgCl (0.87 mL, 1.7 M, 2.1 equivalents) was slowly added under an N2 atmosphere. The mixture was stirred at room temperature for 2 hours, quenched with saturated NH4Cl aqueous solution, and extracted with  (10 mL three times). The combined organic layer was washed with water and brine (20 mL), dried over anhydrous Na2SO4, and concentrated. The crude product was purified by column chromatography to obtain the product (260 mg, 50%) as a white powder. 1 H NMR(400MHz,CD3OD)δ 7.98(s,1H), 7.25(m,5H), 6.23(d,J=18.8Hz,1H), 4.52(m,3H), 4.38(m, 1H), 3.81(m,1H), 3.75(m,1H), 3.48(s,6H), 2.81(m,1H), 1.32(m,18H). [M+H=666.9. [ka]

[0250] Example 17. Preparation of compound 35. [ka]

[0251] Step 1. Preparation of compound 33. AcCl (0.414 g, 1.5 equivalents) was added at 0°C to a mixture of 29 (2.0 g, 1.0 equivalent), DMAP (0.04 g, 0.1 equivalent), pyridine (4 mL), and DCM (20 mL). After stirring at room temperature for 2 hours, TLC showed that the starting materials had been consumed. The mixture was quenched with water, washed with 1 M aqueous HCl, saturated aqueous NaHCO3, and then brine, and dried over anhydrous Na2SO4 to concentrate. The crude product was purified by column chromatography to obtain product 33 (1.73 g, 80.8%) as a white oil. 1 H NMR(400MHz,CDCl3)δ 7.99(s,1H), 7.74(s,1H), 6.20(d,J=20.0Hz,1H), 4.33~4.11(m,4H), 3.50(s,6H), 2.63(s,3H), 2.3(m,1H), 1.26~1.05(m,29H). [M+H] + = 611.9. [ka]

[0252] Step 2. Preparation of compound 34. To a mixture of 33 (1.58 g, 1.0 equivalent) in THF (20 mL), Et3N·3HF (2.1 g, 5 equivalents) and Et3N (2.6 g, 10 equivalents) were added at room temperature. After stirring at room temperature for 1.5 hours, TLC showed that the starting material had been consumed. The mixture was concentrated and purified by column chromatography to obtain 34 (782 mg, 82%) as a white powder. [M+H] + = 369.6. [ka]

[0253] Step 3. Preparation of compound 35. Compound 34 (136 mg, 1.0 equivalent) and PPAL-S (184 mg, 1.1 equivalent) were dissolved in anhydrous THF (3 mL). After cooling the mixture to -5°C, t-BuMgCl was dissolved. (0.5 mL, 1.7 M, 2.1 equivalents) was slowly added under an N2 atmosphere. The mixture was stirred at room temperature for 30 minutes, quenched with saturated NH4Cl aqueous solution, and extracted with  (10 mL three times). The combined organic phase was washed with water and brine (20 mL), dried anhydrous, and concentrated. The crude product was purified by column chromatography (DCM:MeOH = 50:1 to 20:1) to obtain phosphoramidate 35 (150 mg, 63.8%) as a white powder. 1 H NMR(400MHz,CD3OD)δ 7.81(s,1H), 7.35~7.16(m,5H), 6.10(d,J=18.4Hz,1H), 4.87(m,1H), 4.52~4 .46(m,3H), 4.21(m,1H), 3.91~3.87(m,1H), 3.03(s,3H), 1.30~1.13(m,12H). 31 P NMR (160 MHz, CD3OD) δ 3.84. 19 F NMR(376MHz,CD3OD)δ -162.79.

[0254] β-D-2'-deoxy-2'-α-fluoro-2'-β-ethynyl-N 6 Synthesis of substituted-2,6-diaminopurine nucleotides [ka]

[0255] Example 18. β-D-2'-deoxy-2'-α-fluoro-2'-β-ethynyl-N 6 - General pathway to substituted-2,6-diaminopurine nucleotides [ka]

[0256] Step 1. Preparation of compound 36. To a solution of 6-chloroguanosine (100 g, 332 mmol) in pyridine (400 mL), TPDSCl2 (110 mL, 1.05 equivalents) was added dropwise at -5°C to 5°C under an N2 atmosphere. After stirring at this temperature for 2 hours, TLC showed that the starting material had been consumed. After adding DCM (600 mL), TMSCl (85 mL, 2 equivalents) was added dropwise at 0°C to 5°C. After stirring at this temperature for 2 hours, TLC showed that the intermediate had been consumed.

[0257] Isobutyryl chloride was added dropwise at 0°C to 5°C. After stirring at that temperature for 2 hours, TLC showed that the intermediate had been consumed. Water was added, and the contents were extracted by DCM. Subsequently, The organic phase was washed with 0.5N HCl to remove pyridine. The contents were washed to adjust the pH to 5-6, and then pTSA·H2O (9.2g, 484.5 mmol) was added at 0°C-5°C. After stirring at that temperature for 1 hour, TLC showed that the intermediate had been consumed. Water was then added, and the organic phase was washed with water, saturated NaHCO3 water, and brine. After drying with Na2SO4, the solvent was removed under vacuum. The residue was then purified by column chromatography (PE / EA = 100 to 10 / 1) to obtain the product (82g, 40%) as a bright yellow solid. 1 H NMR(400MHz,DMSO-d6)δ 10.88(s,1H), 8.55(s,1H), 5.91(d,J=1.6Hz,1H), 5.53(d,J=4.6Hz,1H), 4.72~4.58(m,2H), 4.16(dd,J=12.4,4.8Hz,1H), 4.00(ddd,J= 7.7,4.8,2.6Hz,1H), 3.93(dd,J=12.4,2.7Hz,1H), 2.78(h,J=6.9Hz,1H), 1.26~1.12(m,3H), 1.10(d,J=6.7Hz,6H), 1.09~0.88(m,24H). [ka]

[0258] Step 2. Preparation of compound 37. Desmartin periodinane was added to a 100 mL solution of 36 (10.0 g, 16.3 mmol) in DCM, and the reaction was stirred for 12 hours. TLC showed that the starting material had been consumed. The reaction mixture was then diluted with 200 mL of DCM and washed with saturated Na2S2O3 water and brine. The organic phase was then dried over Na2SO4 and concentrated to obtain crude 37 (12 g) as a bright yellow solid. Crude 53 can be used directly in the next step without purification. [ka]

[0259] Step 3. Preparation of compound 38. To a solution of ethynyltrimethylsilane (18.6 mL, 142.7 mmol) in THF (240 mL), n-BuLi (46 mL, 2.5 M, 115.0 mmol) was added dropwise under an N2 atmosphere at -15°C to -20°C. After stirring for 30 minutes, the reaction mixture was cooled to -70°C, and 37 (crude, 16.3 mmol) from THF (60 mL) was added at that temperature. The contents were then warmed to 0°C. TLC showed that the starting material had been consumed. Saturated NH4Cl water was added, and the reaction mixture was extracted three times with EA (100 mL). After combining the organic phases, the mixture was washed with brine and then further dried with Na2SO4. After concentration under vacuum, the residue was purified by column chromatography (PE / EA = 100 → 10 / 1) to obtain a bright yellow solid (6 0.0g and 52% were obtained. [ka]

[0260] Step 4. Preparation of compound 39. Pyridine (4.2 mL, 52.9 mmol) was added to a solution of 38 (6.0 g, 8.4 mmol) in DCM (240 mL) under an N2 atmosphere. The reaction mixture was cooled to -70°C, and DAST (12 mL, 90.4 mmol) was added. The contents were then warmed to -30°C. TLC showed that the starting materials had been consumed. The reaction mixture was poured into saturated NaHCO3 water and extracted with DCM (200 mL). The organic phase was washed with brine and dried over Na2SO4. After concentration under vacuum, the residue was purified by column chromatography (PE / EA = 100 → 10 / 1) to obtain a bright yellow solid (3.8 g, 63%). [ka]

[0261] Step 5. Preparation of compound 40. AcOH (1.3g, 22 mmol) and TBAF (4.2g, 15.9 mmol) were added to a 39 (3.8g, 5.3 mmol) THF (120 mL) solution at room temperature. The reaction mixture was stirred at room temperature for 30 minutes. TLC showed that the starting materials were consumed. After concentration under vacuum, the residue was purified by column chromatography (EA) to obtain the product (2.0g, 95%) as a white solid. [ka]

[0262] General procedures for amino substitution and deprotection: Add methanol or the corresponding amine aqueous solution (as a hydrochloride, free base or salt and DIEA) to a 20 mL solution of 40 (350 mg, 0.88 mmol) of dioxane at room temperature. ) was added. The contents were stirred at room temperature for 1 to 12 hours. TLC showed that the starting material had been consumed. After concentration under vacuum, the residue was used directly in the next step without purification. The residue mentioned above was dissolved in methanol (10 mL). NaOH water (2.5 N, 10 mL) was added. After stirring overnight at room temperature, TLC showed that the starting material had been consumed. The pH of the contents was adjusted to 7-8 with 1 N HCl. The solution was concentrated and purified by column chromatography (DCM / MeOH = 100 → 20 / 1) to obtain the product as a grayish-white solid (yield: 40%-80% for 2 steps). Table 1 shows the structures of compounds 57-63, and their corresponding mass spectra, and the properties of each compound. 1 This shows the 1H NMR spectrum.

[0263] [Table 3]

[0264] Example 19. Preparation of isopropyl((((R,S)-(2R,3R,4R,5R)-5-(2-amino-6-dimethylamino-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-ethynyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate [ka] i) Isopropyl((R,S)-(pentafluorophenoxy)-phenoxy-phosphoryl)-L-alaninate, tBuMgCl, THF, 0℃.

[0265] Step 1. Preparation of isopropyl((((R,S)-(2R,3R,4R,5R)-5-(2-amino-6-dimethylamino-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-ethynyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate. To a solution of compound 41 (30 mg, 0.09 mmol) in dry THF (2 mL) at 0°C, tert-butylmagnesium chloride (1.0 M in THF, 125 μL, 0.13 mmol) was added dropwise over 10 minutes. The reaction mixture was stirred at 0°C for 15 minutes, and then stirred at room temperature for another 15 minutes. The reaction mixture was cooled to 0°C, and a solution of isopropyl((R,S)-(pentafluorophenoxy)-phenoxy-phosphoryl)-L-alaninate (49 mg, 0.11 mmol) dissolved in dry THF (2 mL) was added dropwise over 10 minutes. The reaction mixture was stirred at 0°C for 30 minutes and at room temperature for 18 hours. The reaction mixture was quenched with saturated NH4Cl aqueous solution (4 mL) and extracted with ethyl acetate (5 mL three times). The combined organic matter was dried over Na2SO4 and concentrated. The residue was purified by column chromatography (DCM / MeOH gradient from 100:0 to 90:10) to obtain the product (a mixture of two diastereoisomers, 12 mg, 0.02 mmol, 24%) as a white solid. 1 H NMR(300MHz,CD3OD)δ 7.79(s,0.45H), 7.77(s,0.55H), 7.36~7.14(m,5H), 6.28(d,J=17.4Hz) and 6.26(d,J=17.5Hz,1H), 5.00~4.44(m ,5H), 4.23~4.16(m,1H), 3.69~3.81(m,1H), 3.42(bs,3H), 3.40(bs,3H), 1.32~1.26(m,3H), 1.20~1.15(m,6H). 31 P NMR (121MHz, CD3OD) δ 4.04(s), 3.98(s). C 26 H 34 FN7O7P[M+H] + MS(ESI)m / z calculated value: 606.2; measured value: 606.2.

[0266] Example 20. Preparation of isopropyl((((R,S)-(2R,3R,4R,5R)-5-(2-amino-6-methylamino-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-ethynyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate. [ka] i) Isopropyl((R,S)-(pentafluorophenoxy)-phenoxy-phosphoryl)-L-alaninate, tBuMgCl, THF, 0℃.

[0267] Step 1. Isopropyl((((R,S)-(2R,3R,4R,5R)-5-(2-Amyl Preparation of no-6-methylamino-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-ethynyltetrahydrofuran-2-yl)methoxy)phenoxy-phosphoryl)-L-alaninate. To a solution of compound 42 (30 mg, 0.09 mmol) in dry THF (2 mL) at 0°C, tert-butylmagnesium chloride (1.0 M in THF, 125 μL, 0.13 mmol) was added dropwise over 10 minutes. The reaction mixture was stirred at 0°C for 15 minutes, and then stirred at room temperature for another 15 minutes. The reaction mixture was cooled to 0°C, and a solution of isopropyl((R,S)-(pentafluorophenoxy)-phenoxy-phosphoryl)-L-alaninate (49 mg, 0.11 mmol) dissolved in dry THF (2 mL) was added dropwise over 10 minutes. The reaction mixture was stirred at 0°C for 30 minutes and at room temperature for 18 hours. The reaction mixture was quenched with saturated NH4Cl aqueous solution (4 mL) and extracted with ethyl acetate (5 mL three times). The combined organic matter was dried over Na2SO4 and concentrated. The residue was purified by column chromatography (DCM / MeOH gradient from 100:0 to 90:10) to obtain the product (a mixture of two diastereoisomers, 9 mg, 0.02 mmol, 18%) as a white solid. 1 H NMR(300MHz,CD3OD)δ 7.81,7.79(0.9s+0.1s,1H), 7.36~7.14(m,5H), 6.26(d,J=17.4Hz,0.1H) and 6.24(d,J=17.4Hz,0.9H), 4.93~4.89(overlapping with H2O, m,1H), 4.80~4.78(m,1H), 4.53~4.49(m,2H), 4.21~4.18(m,1H), 3.95~3.84(m,1H), 3.23~3.20(m,1H), 3.04(bs,1H), 1.31~1.14(m,9H).31 P NMR (121MHz, CD3OD) δ 4.06(s), 3.97(s). C 25 H 32 FN7O7P[M+H] + MS(ESI)m / z calculated value: 592.2; measured value: 592.2.

[0268] Example 21. Preparation of isopropyl((((R,S)-(2R,3R,4R,5R)-5-(2-amino-6-(N-methylcyclopropylamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-ethynyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate [ka] i) Isopropyl((R,S)-(pentafluorophenoxy)-phenoxy-phosphoryl)-L-alaninate, tBuMgCl, THF, 0℃.

[0269] Step 1. Preparation of isopropyl((((R,S)-(2R,3R,4R,5R)-5-(2-amino-6-(N-methylcyclopropylamino)-9H-purine-9-yl)-4-fluoro-3-hydroxy-4-ethynyltetrahydrofuran-2-yl)methoxy)-phenoxy-phosphoryl)-L-alaninate. To a solution of compound 43 (40 mg, 0.11 mmol) in dry THF (2 mL) at 0°C, tert-butylmagnesium chloride (1.0 M in THF, 160 μL, 0.16 mmol) was added dropwise over 10 minutes. The reaction mixture was stirred at 0°C for 15 minutes, and then stirred at room temperature for another 15 minutes. The reaction mixture was cooled to 0°C, and a solution of isopropyl((R,S)-(pentafluorophenoxy)-phenoxy-phosphoryl)-L-alaninate (55 mg, 0.12 mmol) dissolved in dry THF (2 mL) was added dropwise over 10 minutes. Reaction mixture The mixture was stirred at 0°C for 30 minutes and at room temperature for 18 hours. The reaction product was quenched with saturated NH4Cl aqueous solution (4 mL) and extracted with  (5 mL three times). The combined organic matter was dried over Na2SO4 and concentrated. The residue was purified by column chromatography (DCM / MeOH gradient from 100:0 to 90:10) to obtain the product (a mixture of two diastereoisomers, 18 mg, 0.03 mmol, 26%) as a white solid. 1 1H NMR (300MHz, CD3OD)δ 7.84,7.82(s+s,1H), 7.35~7.14(m,5H), 6.30(d,J=17.4Hz) and 6.26(d,J=1 7.6Hz,1H), 4.99~4.89(overlaps with H2O, m,1H), 4.82~4.69(m,1H), 4.59~4.46(m,2 H), 4.21(m,1H), 3.96~3.82(m,1H), 3.24~3.22(m,1H), 3.17~3.11(m,1H)1 .31~1.26(m,3H), 1.20~1.15(m,6H), 0.93~0.89(m,2H), 0.75~0.68(m,2H). 31 P NMR (121MHz, CD3OD) δ 4.06(s), 3.98(s). C 28 H 36 FN7O7P[M+H] + MS(ESI)m / z calculated value: 632.2; measured value: 632.2.

[0270] Example 22. Preparation of PPAL-S [ka]

[0271] Step 1. Preparation of racemic PPAL To a stirred solution of phenyldichlorophosphate (250 g) in  (800 mL), isopropyl L-alaninate (200 g) in triethylamine (120 g) was added at -10°C. The reaction mixture was stirred at -10°C for 1 hour. Compound 2,3,4,5,6-pentafluorophenol (220 g) in triethylamine (120 g) and  (400 mL) was added at -5°C, and the mixture was stirred at that temperature for 0.5 hours. The reaction mixture was warmed to 25°C and stirred at room temperature for 2 hours. The solution was filtered and washed twice with  (200 mL), and the combined organic phase was evaporated under vacuum to obtain solid PPAL-RS (racemic mixture).

[0272] Step 2. Preparation of PPAL-RS To a stirred solution of PPAL-RS in SiO (200 mL) and n-heptane (1.4 L), 10.1 g of 2,3,4,5,6-pentafluorophenol in triethylamine (6 g) was added, and stirring was continued for approximately 4 to 8 hours. After the solid R isomer content decreased to less than 0.5%, the solid was filtered. The solid was dissolved in SiO (4 L), washed with water (twice with 100 mL) and brine (1 L), dried over anhydrous Na2SO4, and filtered. The solvent was removed under vacuum to obtain PPAL-S (350 g). 1 H NMR(400MHz,DMSO-d6)δ=7.42~7.40(m,2H), 7.24~7.22(m,3H), 6.87(dd,J=14.1,9.9Hz,1H), 4. 90~4.84(m,1H), 3.94~3.88(m,1H), 1.27(dd,J=7.1,1.1Hz,3H), 1.15(dd,J=6.2,1.2Hz,6H)ppm. 13 P NMR (160MHz, DMSO-d6) δ=0.37ppm.

[0273] Example 23. Preparation of PPAL-R [ka]

[0274] Phenyl dichlorophosphate (189.6 g, 0.90 mol) and anhydrous ethyl acetate (750 mL) were added to a three-necked round-bottom flask fitted with a mechanical stirrer. The solution was cooled to -10°C under a nitrogen atmosphere. Isopropyl L-alaninate (118 g, 0.90 mmol) and triethylamine (100 g, 1.1 equivalents) were added to the above solution. A pre-cooled (below 10°C) mixture of 2,3,4,5,6-pentafluorophenol (165 g, 1 equivalent) and triethylamine (90.5 g, 1 equivalent) in ethyl acetate (300 mL) was added to the above mixture at -5°C using an addition funnel, and the resulting mixture was stirred for 1 hour between 20°C and 25°C. The white precipitate (TEA·HCl) was removed by filtration, and the mixture was rinsed with ethyl acetate. The filtrate was concentrated under reduced pressure to obtain approximately 280 g of PPAL-RS (S / R=1 / 1) as a white solid. PPAL-RS (280g) was ground in 300mL of heptane / siRNA (20:1) at room temperature for 5 minutes. The white suspension was filtered, and the solid was rinsed with a heptane / siRNA (20:1) mixture. The filtrate was cooled to 8°C, and the solid was collected by filtration. Crude PPAL-R (10g) was obtained with 95% chiral purity. The crude product was purified according to the above procedure. PPAL-R (5g) was obtained with 98% NLT chiral purity. 1 H NMR(400MHz,DMSO-d6)δ=7.43~7.39(m,2H), 7.27~7.22(m,3H), 6.87(dd,J=14.1,9.9Hz,1H), 4 .89~4.85(m,1H), 3.95~3.90(m,1H), 1.27(dd,J=7.1,1.1Hz,3H), 1.14(dd,J=6.2,1.2Hz,6H). 13 P NMR (160MHz, DMSO-d6) δ=0.35.

[0275] Example 24: Preparation of compound 52. [ka]

[0276] Step 1. Preparation of compound 49. A 40% aqueous solution of CH3NH2 (16.2 mmol) was added to a solution of 48 (1.81 g, 3.23 mmol) of dioxane (18 mL). The reaction mixture was stirred at 40°C for 2 hours. The mixture was concentrated, diluted with SiO2 (50 mL), and washed with water and brine. The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated to obtain a white solid 49 (1.66 g, 92%). [ka]

[0277] Step 2. Preparation of compound 50. Pentyl chloroformate (547 mg, 3.63 mmol) was slowly added at 0°C to a solution of 49 (1.34 g, 2.42 mmol) and 1-methylimidazole (794 mg, 9.68 mmol) in DCM (14 mL). The reaction mixture was stirred overnight at room temperature. The mixture was concentrated and purified by column chromatography (PE:SiO = 5:1 to 2:1) to obtain 50 (1.01 g, 62%) as a white solid. 1 H NMR(400MHz,DMSO)δ 7.96(s,1H), 6.73(s,1H), 6.06~6.10(d,J=16.0Hz,1H), 4.09~4.30(m,2H), 3.97~4 .09(m,4H), 3.28(s,3H), 1.39~1.46(m,2H), 1.0~1.2(m,35H), 0.73~0.76(t,J=8.0H z,3H). [ka]

[0278] Step 3. Preparation of compound 51. To a 50 (1.00 g, 1.5 mmol) THF (11 mL) solution, Et3N (2.0 mL, 15 mmol) and Et3N.3HF (1.21 g, 7.5 mmol) were added at 0°C. The reaction mixture was stirred at room temperature for 1.5 hours. The mixture was concentrated and purified by column chromatography (MeOH:CH2Cl2 = 50:1) to obtain 75 (460 mg, 72.2%) as a white powder. [ka]

[0279] Step 4. Preparation of compound 52. To a solution of 51 (460 mg, 1.08 mmol) and PPAL-S (538 mg, 1.19 mmol) in anhydrous THF (9 mL), t-BuMgCl (2.27 mmol) was slowly added under N2 conditions at 5°C to 10°C. The reaction mixture was stirred at room temperature for 40 minutes. The mixture was quenched with saturated NH4Cl aqueous solution, extracted with Â, washed with 5% K2CO3 aqueous solution and brine, dried over anhydrous Na2SO4, filtered, and concentrated. The crude product was purified by column chromatography (CH2Cl2:MeOH = 15:1) to obtain 52 (280 mg, 37.3%) as a white powder. 1 1H NMR (400MHz, DMSO) δ 8.12(s,1H), 7.34~7.38(m,2H), 7.18~7.23(m,3H), 6.74(s,2H), 6.11~ 6.16(d,J=16.0Hz,1H), 5.99~6.05(m,1H), 5.84(m,1H), 4.77~4.81(m, 1H), 4.30~4.41(m,3H), 4.03~4.11(m,3H), 3.78~3.80(m,1H), 3.3(s,3 H), 1.44~1.51(m,2H), 1.00~1.21(m,16H), 0.76~0.80(t,J=8.0Hz,3H). [M+H] + = 696.6.

[0280] Example 25: Preparation of compound 56. [ka]

[0281] Step 1. Preparation of compound 48. TIPDSCl2 (1.5 equivalents) was added to a 30 mL solution of pyridine (600 mg, 1 equivalent) at 0°C. The resulting solution was left at room temperature for 2 hours. The mixture was quenched with ice water and extracted with RINKAN. The organic layer was washed with 1 M aqueous HCl, saturated sodium bicarbonate solution, and saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain the crude residue. The residue was purified by chromatography (MeOH:CH2Cl2 = 1:50) to obtain 48 (998 mg, 94.4%) as a white solid foam. [ka]

[0282] Step 2. Preparation of compound 53. A mixture of 48 (800 mg, 1 equivalent), pyridine (3.2 mL), and DMAP (34.9 mg, 0.2 equivalents) in DCM (20 mL) was stirred at room temperature. N-amyl chloroformate (3.2 mL) was added dropwise at 0°C, and the mixture was stirred at room temperature for 1 day. The organic layer was washed with 1 M aqueous HCl, saturated sodium bicarbonate solution, and saturated sodium chloride solution, and anhydrous sodium sulfate was used. The solution was dried with aluminium and evaporated under vacuum. The residue was purified by chromatography on silica gel (MeOH:CH2Cl2=1:50) to obtain 53 (255 mg, 26%) as a white solid foam. [ka]

[0283] Step 3. Preparation of compound 54. To a 10 mL solution of 1,4-dioxane (270 mg, 1 equivalent) of 53, 40% aqueous CH3NH2 solution (225.7 mg, 5 equivalents) was added dropwise. The mixture was stirred at room temperature for 2 hours and then concentrated under vacuum. The residue was subjected to chromatography on silica gel (methanol:dichloromethane = 1:40) to obtain 54 (220 mg, 81.7%) as a white solid foam. [ka]

[0284] Step 4. Preparation of compound 55. Triethylamine (1011.9 mg, 10 equivalents) and Et3N·3HF (806.05 mg, 5 equivalents) were added to 54 (668 mg, 1 equivalent) of THF (10 mL) ice-cold solution, and the mixture was stirred at room temperature for 2 hours. The mixture was concentrated and subjected to chromatography on silica gel (MeOH:CH2Cl2 = 1:30) to obtain 55 (492 mg, 84%) as a white solid foam. [ka]

[0285] Step 5. Preparation of compound 56. A mixture of 55 (113 mg, 1 equivalent) and PPAL-S (120 mg, 1 equivalent) in THF (4 mL) was dropwise added with 1.7 M t-BuMgCl in THF (0.327 mL, 2.1 equivalents) at -10°C. The mixture was stirred at room temperature for 1 hour and then quenched with saturated NH4Cl aqueous solution. The aqueous phase was extracted with ethyl acetate, the organic phase was washed with brine, dried, and concentrated to obtain the crude residue. The residue was subjected to flash chromatography to obtain 56 (126 mg, 68.5%) as a white solid. 1 H NMR(400MHz,DMSO)δ 8.00(s,1H), 7.10~7.45(m,5H), 6.15~6.20(d,J=20.0Hz,1H), 5.00~5.25(s,1H), 4.80~4.86(m,1H), 4.45~4.70(m,2H), 4.12~4.19(m,3H), 3.80~3.85(m,1H), 3.04(s,3H), 1.60~1.75(m,2H), 1.10~1.40(m,16H), 0.76~0.80(t,J=8.0Hz,3H). 31 P NMR(160MHz,DMSO)δ 3.57.[M+H] + = 696.5.

[0286] Example 26: Preparation of compound 60. [ka] JPEG0007893504000173.jpg42169

[0287] Step 1. Preparation of compound 57. To a 100 mL solution of 6 (20 g, 1 equivalent) of CH3CN, imidazole (16.6 g) and TIPDSCl2 (28.9 g, 1.5 equivalents) were added sequentially at 5 ± 5 °C. The resulting solution was left at room temperature for 4 hours. The mixture was quenched with ice water and extracted with HCl. The organic layer was washed with water, saturated sodium bicarbonate solution, and saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain the crude residue (32 g). [ka]

[0288] Step 2. Preparation of compound 58. To a 57 (9.8g, 1 equivalent) THF (4mL) solution, add THF (50mL, 4.8 equivalents) 1.7 M t-BuMgCl was added dropwise at 0°C to 5°C. The mixture was stirred at room temperature for 0.5 hours, and n-amyl chloroformate (2.7 g, 1.05 equivalents) was slowly added. The mixture was stirred at 0°C to 5°C for 3 to 4 hours. The mixture was quenched with saturated NH4Cl aqueous solution. The aqueous phase was extracted with ELISA (200 mL), the organic phase was washed with brine, dried, and concentrated to obtain 58 (10.7 g) of oil. [ka]

[0289] Step 3. Preparation of compound 59. Triethylamine (10.119 g) and Et3N·3HF (8.6 g, 5 equivalents) were added to a 100 mL ice-cold THF solution of 58 (7.3 g, 1 equivalent), and the mixture was stirred at room temperature for 1 hour. The mixture was concentrated and subjected to chromatography on silica gel (MeOH:CH2Cl2 = 1:30) to obtain 59 (4.3 g, 91%) as a white solid. [ka]

[0290] Step 4. Preparation of compound 60. A mixture of 59 (2 g, 1 equivalent) and PPAL-S (2.3 g, 1.1 equivalents) in THF (40 mL) was dropwise added with 1.7 M t-BuMgCl in THF (5.6 mL, 2.1 equivalents) at -5°C. The mixture was stirred at -20±5°C for 1 hour and then quenched with saturated NH4Cl aqueous solution. The aqueous phase was extracted with ethyl acetate, the organic phase was washed with brine, dried, and concentrated to obtain the crude residue. The residue was subjected to flash chromatography to obtain 60 (1.5 g, 47%) as a white solid. 1 H NMR(400MHz,CD3OD)δ 7.9(s,1H), 7.1~7.2(m,5H), 6.2(d,J=20Hz,1H), 5.1(br,1H), 4.84(m,1H), 4.49(m,,2H), 4.16(m,1H), 4 .13(m,2H), 3.86(m,1H), 3.45(br,6H), 1.70(m,2H), 1.26(m,4H), 1.20(m,6H), 1.14(m,6H), 0.93(m,3H). [M+H] + = 710.5.

[0291] Biological data Example 27. Assay methodology and additional biological data Antiviral efficacy (EC 50 ) and cytotoxicity (TC 50 To enable parallel determination of ), Huh-7 luc / neo ET cells carrying a disisstronic HCV genotype 1b luciferase reporter replicon were placed in a double-row 96-well plate in 7.5 × 10⁶ units. 3 Cells were seeded at a concentration of cells / ml. The plates were incubated for 24 hours prior to the addition of the compound. Six consecutive samples of the test substance were added. A 3.2-fold (one half log: 1 / 2 log) dilution series (high test concentration of 100.0 μM or 1.0 μM) and human interferon-alpha 2b (high test concentration of 10.0 U / ml) were prepared in cell culture medium, and each dilution was added to cells cultured in a triple well. Six wells in the test plate were used as an untreated control with only culture medium. After 72 hours of incubation in the presence of the compounds, one plate was used to determine cytotoxicity by staining with XTT, and the others were used to determine antiviral efficacy by assessing luciferase reporter activity. TC 50 and EC 50 To determine the value, cytotoxicity and potency data were collected and imported into a customized Excel workbook. The data for compounds of formulas I to VII are shown in Table 7 below. Furthermore, Figure 2 shows the HCV replication inhibition curves for compound 5-2 and sofosbuvir. As can be seen from Figure 2, compound 5-2 is EC 50 While it has =4nM, sofosbuvir has EC 50 It has a concentration of 53 nM. The y-axis is the percentage relative to the viral control, and the x-axis is the concentration of the drug in μM units. Figure 3 shows the HCV replication inhibition curves for compound 25 and sofosbuvir. Compound 25 is EC 50 It has =4nM, and sofosbuvir is EC 50 It has a concentration of 53 nM. The y-axis represents the percentage relative to the viral control, and the x-axis represents the drug concentration in μM units. Figure 4 shows an intraassay comparison of anti-HCV activity for compound 5-2, compound 25, compound 27, and sofosbuvir. The y-axis represents the percentage relative to the viral control, and the x-axis represents the drug concentration in μM units.

[0292] The relative replication susceptibility of HCV genotypes to test compounds was determined using HCV genotypes derived from various patients, including wild-type and resistance-related variants. Replicon resistance test vectors (RTVs) containing the NS5B genomic region were prepared using viral RNA isolated from the plasma of HCV patients. Each NS5B region was amplified by reverse transcription polymerase chain reaction, cloned into HCV replicon RTVs, and then transferred into Huh-7 cells by electroporation. After 72-96 hour incubation in and without serially diluted test compounds, viral replication was measured by luciferase activity, and the 50% inhibitory concentration (IC) was measured. 50 The value was determined.

[0293] Table 2 shows IC25, Compound 27, Compound 5-2, and sofosbuvir against various clinical isolates, including wild-type and resistance-associated mutants. 50 and IC 95 Report the value of [the value].

[0294] All compounds were significantly more effective against HCV replication than sofosbuvir, and none of compounds 25, 27, and 5-2 showed any evidence of cross-resistance to the L159F, L159F, S282T, and C316N mutants.

[0295] [Table 4]

[0296] To determine the susceptibility of the HCV wild-type S282T mutant to the test compound, A transient transfect assay was performed. Huh-7 cells were electroporated from the T7 promoter in the presence of RNA transcribed from wild-type or S282T HCV replicon plasmids. Transfected cells were placed in Dulbecco's modified Eagle medium at a rate of 7.5 × 10⁶ per well. 3Cells were seeded in 96-well plates. After 24 hours of incubation, the medium was removed and replaced with fresh medium containing either no test compound or varying concentrations of the test compound. After a further 96 hours of incubation, anti-HCV activity was measured by the luciferase endpoint using the Britlite® Plus Luminescence Reporter Gene Kit (Perkin Elmer, Shelton, Connecticut). Duplicate plates were processed and incubated in parallel for evaluation of cytotoxicity by staining with tetrazolium dye XTT.

[0297] Table 3 shows the IC25, compound 27, compound 5-2, and sofosbuvir IC255, compound 27, compound 5-2, and sofosbuvir IC255 for HCV wild-type and S282T replicons. 50 and IC 95 Report the value of [the value].

[0298] All compounds were significantly more effective against HCV replication than sofosbuvir, and none of the compounds 25, 27, or 5-2 showed any evidence of cross-resistance to the S282T variant.

[0299] [Table 5]

[0300] The stability of selected compounds in fresh human whole blood and human liver S9 fraction was determined in incubations containing 10 μM of the test compound. After incubations of 0, 30, and 60 minutes, and up to 120 minutes, aliquots were removed and immediately extracted with 3 volumes of ice-cold methanol / acetonitrile (1:1, volume / volume). The extracts were centrifuged, and the supernatant was analyzed by LC-MS / MS for the concentrations of the unchanged test compound and potential metabolites.

[0301] Figure 5 shows the excellent stability of compound 5-2 and all 2-amino derivatives in human blood.

[0302] Interestingly, Figure 6 shows the 2'-deoxy-2'-α-fluoro-2'-β-methyl-N2 from the human liver S9 fraction. 2 -methyl-N 6 -Methyl-2,6-diaminopurine nucleoside phosphoramidate 2'-deoxy-2'-α-fluoro-2'-β-methyl-N 6 It exhibits in vitro, time-dependent dealkylation to methyl-2,6-diaminopurine nucleoside phosphoramidate. Furthermore, compared to compound 5-2 and other 2-amino derivatives, it reacts with unexpected, faster, and more broad proportions in the human liver S9 fraction. Cleavage of the carbamate portion was observed (Figure 7).

[0303] Example 28. HCV(gt1b)NS5B polymerase assay Inhibition of HCV(gt1b)NS5B polymerase was determined by measuring de novo polymerization in a reaction mixture containing diluted TA, in vitro transcribed viral RNA complementary to the HCV(-) chain 3'UTR region, polymerase, radioisotope-recognizing ribonucleotide, 250 μM non-competitive rNTP, and 1 μM competitive rNTP, repeated three times. 50% inhibition (IC) 50 The TA concentration that produced the desired result was determined from the obtained inhibition curve.

[0304] Example 29. Human bone marrow primordial cell assay Fresh human bone marrow progenitor cells (Invitrogen) are suspended in either BFU-E or GM-CSF-specific culture medium, and in a 6-well plate, 10 times the dilution series of triple TA. 5 Cells were added in the well. After 14 days of incubation, CC 50 Colony count was used to determine the value. BFU-E colonies were identified using benzidine technology.

[0305] Compounds 25, 27, and 5-2 do not exhibit cytotoxicity to bone marrow stem cells in vitro.

[0306] Example 30. iPS cardiomyocyte assay iPS cardiomyocytes (Cellular Dynamics) 1.5 × 10⁶ per well 4 Microphone in a cell Cells were seeded in Lottle plates. After 48 hours of incubation, the cells were washed and maintenance medium containing serially diluted TA was added in three sets. After a further 3 days of incubation, cell viability was measured by staining with XTT and CC. 50 The value was calculated.

[0307] Compounds 25, 27, and 5-2 do not exhibit cytotoxicity to iPS cardiomyocytes in vitro.

[0308] Example 31. Human DNA polymerase assay Inhibition of human DNA polymerases α, β, and γ (CHIMERx) was observed with serially diluted TA, 0.05 mM dCTP, dTTP, and dATP, 10 μCi[ 32 The reaction was repeated three times in a reaction mixture of [P]-α-dGTP (800 Ci / mmol), 20 μg of activated calf thymus DNA, and additional reagents specific to each polymerase. After a 30-minute incubation, [α- 32 The integration of P]-GTP was measured, and the resulting incubation curve was used for IC 50 It was used to calculate the value.

[0309] Not only the triphot β-D-2'-deoxy-2'-α-fluoro-2'-β-methylguanine triphot, but also the triphot analogs of compound 25, compound 27, and compound 5-2 do not inhibit human DNA polymerase α, β, or γ.

[0310] Example 32. Human hepatocyte co-culture Following a procedure established by Hepregen, micropatterned human hepatocyte co-cultures (HepatoPac®, Hepregen Corporation) were prepared by seeding cryopreserved female human hepatocytes (single donor) and 3T3 J2 mouse fibroblasts in microtiter plates. ALT leakage, urea production, albumin secretion, and cellular ATP content were measured three times to assess cytotoxicity and hepatocyte health. The culture medium was replaced every two or three days for 16 days with fresh medium containing TA and test substances (0 μM, 1 μM, 10 μM, or 30 μM). Used culture media were assayed for ALT and urea content on days 2, 5, 7, 9, 12, 16, and 21, and for albumin content on days 2, 5, 7, and 9. Cellular ATP levels were measured on days 9 and 21. Measurements were taken on day 1. ATP signaling in a control culture of stromal cells only (murid 3T3 fibroblasts) was subtracted from that of a human HepatoPac co-culture to obtain hepatocyte-specific effects. See Tables 4, 5, and 6 below.

[0311] Compound 5-2, at concentrations up to 30 μM, showed no signs of cytotoxicity, as measured by ALT leakage, albumin secretion, urea production, and cellular ATP content, when incubated in micropatterned co-cultured human hepatocytes for up to 12 days. The slight signs of cytotoxicity detected during extended exposure (up to 21 days of culture) were significantly lower than those observed with sofosbuvir. See Tables 4, 5, and 6 below.

[0312] INX-189 showed high cytotoxicity against human co-cultured hepatocytes, exhibiting decreased albumin secretion and cytotoxicity as early as day 2 in all measurements. Sofosbuvir showed higher cytotoxicity than AT-511 under the same conditions.

[0313] [Table 6]

[0314] [Table 7]

[0315] [Table 8]

[0316] Example 33. Metabolic Studies The metabolism of compounds 25, 27, and 5-2 was investigated at a concentration of 10 μM in fresh primary cultures of human, canine, and mouse hepatocytes. Human cells (XenoTech, mixed sex, collected from 10 donors) were seeded in 6-well plates coated with Matrigel. Hepatocytes derived from male beagle dogs (BioreclamationIVT) and male ICR / CD-1 mice (BioreclamationIVT, 8 donors) were incubated with 10 μM TA in singlets for 2 hours and 4 hours. After 6, 8, or 24 hours, intracellular levels of nucleotide prodrugs and their potential metabolites (prodrugs, monophosphates, triphots, and nucleosides) were quantified by LC-MS / MS. Concentrations below the limit of quantification (1.5 pmol / 10 for prodrugs, monophosphates, and nucleosides) were measured. 6 12 pmol / 10 for cells and triphot phosphates 6 The cells were estimated from the standard curve.

[0317] Compound β-D-2'-deoxy-2'-α-fluoro-2'-β-methyl-guanine triphosphate is the main metabolite of compounds 25, 27, and 5-2 observed in cultured human hepatocytes, and is present in 0.15 μM IC2. 50 It is a potent inhibitor of HCV(gt1b)NS5B polymerase.

[0318] Figure 8 shows the metabolites of 25 major compounds in human liver cells.

[0319] Figure 9 shows the metabolites of 27 major compounds in human liver cells.

[0320] Figure 10 shows the metabolites of the main compound 5-2 in human liver cells.

[0321] Figure 11 shows the activation pathways for compounds 25, 27, and 5-2. As can be seen from the figure, compounds 25, 27, and 5-2 are converted to their corresponding monophosphate analogs and then metabolized to a common MP analog, namely β-D-2'-deoxy-2'-α-fluoro-2'-β-methyl-guanine monophosphate (compound 61). Subsequently, the monophosphate is stepwise phosphorylated to the active triphosphate, namely β-D-2'-deoxy-2'-α-fluoro-2'-β-methyl-guanine triphosphate (compound 62).

[0322] Example 34. Control INX-189 (INX-08189 / BMS-986094) and sofosbuvir were used as controls in the above examples.

[0323] Two of the most potent nucleotide prodrugs, compound 25 and compound 27, produced over 100 μM of nucleotide-controlled cell volume (CC) in Huh-7 cells, human bone marrow stem cells, and human cardiomyocytes. 50 Along with the values, excellent selectivity was demonstrated. In all host cell lines, no inhibition of human DNA polymerase α, β, or γ, activity against other RNA viruses or DNA viruses, and toxicity were observed up to a maximum concentration of 100 μM.

[0324] Table 7 shows EC 50 / EC 95 (μM) and CC 50 This table explains the compounds tested in the HCV replicon assay, along with the (μM) results.

[0325] [Table 9] JPEG0007893504000183.jpg178149JPEG0007893504000184.jpg170150JPEG0007893504000185.jpg165149JPEG00078935040 00186.jpg142150JPEG0007893504000187.jpg163149JPEG0007893504000188.jpg175150JPEG0007893504000189.jpg131150

[0326] The β-D-2'-D-2'-α-fluoro-2'-β-C-substitution-2-modification-N described herein 6 - Substituted purine nucleotides exhibit significant activity against the HCV virus. The compounds according to the present invention are assayed for desired relative activity using conventional assays that are well known and found in the literature.

[0327] For example, the anti-HCV activity and cytotoxicity of the above compounds may be measured using an HCV subgenomic RNA replicon assay system in Huh7 ET cells. (Korba, et al.) See Antiviral Research 2008, 77, 56). Results were obtained using a positive control and 2'-C-Me cytosine {2'-C-Me-C} (Pierra, et al., Journal of Medicinal Chemistry). This can be summarized by comparing it with (2006, 49, 6614).

[0328] Another in vitro assay for anti-hepatitis C virus activity is a U.S. patent by Stuyver, et al., further transferred to Pharmasset, Inc. under U.S. Patent No. 7,718,7 It will be published in issue 90.

[0329] This specification is described with reference to embodiments of the present invention. Those skilled in the art can modify the present invention for desired purposes based on the teachings herein, and such modifications are considered to be within the scope of the invention. Item 1 Formula I below:

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Claims

1. The following formula: A compound of the same, or a pharmaceutically acceptable salt thereof, A pharmaceutical composition for the treatment of a human being having a disease selected from the group consisting of a positive anti-HCV antibody and antigen-positive state, chronic liver inflammation caused by the virus, cirrhosis, chronic or acute hepatitis C, fulminant hepatitis C, chronic persistent hepatitis C, and fatigue caused by anti-HCV.

2. The aforementioned compound, The pharmaceutical composition according to claim 1, or a pharmaceutically acceptable salt thereof.

3. The aforementioned compound, The pharmaceutical composition according to claim 1, or a pharmaceutically acceptable salt thereof.

4. The aforementioned compound, The pharmaceutical composition according to claim 1, or a pharmaceutically acceptable salt thereof.

5. The aforementioned compound, The pharmaceutical composition according to claim 1, or a pharmaceutically acceptable salt thereof.

6. The aforementioned compound, The pharmaceutical composition according to claim 1, or a pharmaceutically acceptable salt thereof.

7. The pharmaceutical composition according to any one of claims 1 to 6, wherein the disease is characterized by being positive for anti-HCV antibodies and antigens.

8. The pharmaceutical composition according to any one of claims 1 to 6, wherein the disease is chronic liver inflammation caused by a virus.

9. The pharmaceutical composition according to any one of claims 1 to 6, wherein the disease is cirrhosis of the liver.

10. The pharmaceutical composition according to any one of claims 1 to 6, wherein the disease is chronic or acute hepatitis C.

11. The pharmaceutical composition according to any one of claims 1 to 6, wherein the disease is fulminant hepatitis C.

12. The pharmaceutical composition according to any one of claims 1 to 6, wherein the disease is chronic persistent hepatitis C.

13. The pharmaceutical composition according to any one of claims 1 to 6, wherein the disease is fatigue caused by anti-HCV.

14. The following formula: A compound of (wherein R4 is a monophosphate, diphosphate, or triphosphate), or a pharmaceutically acceptable salt thereof.

15. The compound according to claim 14, wherein R4 is a monophosphate.

16. The compound according to claim 14, wherein R4 is a diphosphate.

17. The compound according to claim 14, wherein R4 is a triphosphate.

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