EA046452B1 - 2'-FLUORINE-SUBSTITUTED CARBANUCLEOSIDE ANALOGUES FOR ANTIVIRAL TREATMENT - Google Patents

Description

Field of technology

The invention relates generally to compounds with antiviral activity, in particular to nucleosides active against Flaviviridae infections and, in particular, to inhibitors of the RNA-dependent RNA polymerase of the hepatitis C virus.

State of the art

Viruses comprising the family Flaviviridae contain at least three distinct genera, including pestiviruses, flaviviruses and hepaciviruses (Calisher et al., J. Gen Virol., 1993, 70, 37-43). While pestiviruses are the cause of many economically important animal diseases, such as bovine diarrhea virus (BVDV), classical swine fever virus (CSFV, hog cholera), and borderline disease of sheep (BDV), their importance in human diseases is less characterized (Moennig, V., et al., Adv. Res. 1992, 48, 53-98). Flaviviruses are responsible for important human diseases such as dengue fever and yellow fever, while hepaciviruses lead to hepatitis C virus infection in humans. Other important viral infections caused by the Flaviviridae family include West Nile virus (WNV), Japanese encephalitis virus (JEV), tick-borne encephalitis virus, Junjin virus, Murray Valley encephalitis, St. Louis encephalitis, Omsk hemorrhagic fever virus, and Zika virus. Combined infections of the Flaviviridae virus family cause significant mortality, morbidity, and economic losses worldwide. Thus, there is a need to develop effective treatments for Flaviviridaevirus infections.

Hepatitis C virus (HCV) is the leading cause of chronic liver disease worldwide (Boyer, N. et al. J Hepatol. 32:98-112, 2000), and much of the current antiviral research is focusing on developing improved treatments for chronic HCV. -infections in humans (Di Besceglie, A.M. and Bacon, B.R, Scientific American, Oct.: 80-85, (1999); Gordon, C.P., et al., J. Med. Chem. 2005, 48, 1- 20; Maradpour, D. et al., Nat. Rev. 2007, 5(6), 453463). A number of treatments for HVC are reviewed by Bymock et al. in Antiviral Chemistry & Chemotherapy, 11:2; 79-95 (2000).

RNA-dependent RNA polymerase (RdRp) is one of the most well-studied targets for the development of new HCV therapeutic agents. NS5B polymerase is a target for inhibitors in early human clinical trials (Sommadossi, 1, WO 01/90121 A2, US 2004/0006002 A1). These enzymes have been extensively characterized at the biochemical and structural levels with screening to identify selective inhibitors (De Clercq, E. (2001) J. Pharmacol Exp. Ther. 297:1-10; De Clercq, E. (2001) J. Clin. Virol. 22:73-89). Biochemical targets such as NS5B are important in the development of hepatitis C therapies since HCV does not replicate in the laboratory and there are difficulties in developing cell-based assays and preclinical animal systems.

Currently, there are mainly two antiviral compounds, the nucleoside analogue ribavirin, and interferon alpha (α) (IFN), which are used to treat chronic HCV infection in humans. Ribavirin alone is not effective in reducing viral RNA levels, has significant toxicity, and is known to cause anemia. The combination of interferon and ribavirin has been reported to be effective in the treatment of chronic hepatitis C (Scott, L. J., et al. Drugs 2002, 62, 507-556), but less than half of patients infected with certain genotypes showed consistent benefit with such treatment. Other patent applications disclosing the use of nucleoside analogues for the treatment of hepatitis C virus include WO 01/32153, WO 01/60315, WO 02/057425, WO 02/057287, WO 02/032920, WO 02/18404, WO 04/046331, WO2008/089105 and WO2008/141079, but additional treatment for HCV infection has not yet become available to patients.

Virological treatment of patients with chronic HCV infection is difficult to achieve due to the enormous amount of daily virus production in chronically infected patients and the high spontaneous variability of the HCV virus (Neumann, et al., Science 1998, 282, 103-7; Fukimoto, et al., Hepatology, 1996, 24, 1351-4; Domingo, et al., Gene, 1985, 40, 1-8; Martell, et al., J. Virol 1992, 66, 3225-9). Experimental antiviral nucleoside analogs have been shown to cause viable mutations of the HCV virus both in vivo and in vitro (Migliaccio, et al., J. Biol. Chem. 2003, 926; Carroll, et al., Antimicrobial Agents Chemotherapy 2009, 926; Brown , A. B., Expert Opin. Investig. Drugs 2009, 18, 709-725). Thus, drugs with improved antiviral properties, in particular, increased activity against resistant strains of the virus; improved oral bioavailability; fewer unwanted side effects and an extended effective half-life in vivo (De Francesco, R. et al. (2003) Antiviral Research 58:1-16) are urgently needed.

Some ribosides from the nucleic bases pyrrolo[1,2-G][1,2,4]triazine, imidazo[1,5G||1.2.4|triazine. imidazo[1,2-G][1,2,4]triazine and [1,2,4]triazole[4,3-1][1,2,4]triazine were disclosed in Carbohydrate Research 2001, 331(1 ), 77-82; Nucleosides & Nucleotides (1996), 15(1-3), 793-807; Tetrahedron Letters (1994), 35(30), 5339-42; Heterocycles (1992), 34(3), 569-74; J. Chem. Soc. Perkin Trans. 1 1985, 3, 621-30; J. Chem. Soc. Perkin Trans. 1 1984, 2, 229-38; WO 2000056734; Organic Letters (2001), 3(6), 839-842; J. Chem. Soc. Perkin Trans. 1 1999, 20, 2929-2936; and J. Med. Chem. 1986, 29(11), 2231-5. However, these compounds have not been disclosed as useful for the treatment of HCV.

- 1 046452

Ribosides pyrrolo[1,2-£][1,2,4]triazinyl, imidazo[1,5-£][1,2,4]triazinyl, imidazo[1,2А1,2,4]triazinyl and [1, 2,4]triazole[4,3-ph][1,2,4]triazinyl nucleobases with antiviral, anti-HCV and anti-RdRp activities were disclosed by Babu, Y. S., WO2008/089105 and WO2008/141079; Cho, et al., WO2009/132123 and Francom, et al. WO2010/002877. Butler, et al., WO2009/132135, disclosed antiviral pyrrolo[1,2-£][1,2,4]triazinyl, imidazo[1,5-A1,2,4]triazinyl, imidazo[1,2-A1 ,2,4]triazinyl and [1,2,4]triazole[4,3-£][1,2,4]triazinyl nucleosides, in which the sugar is substituted in the 1'-position of the nucleoside.

The essence of the invention

Compounds that inhibit viruses of the Flaviviridae family are provided. The invention also includes compounds of formula V that inhibit viral nucleic acid polymerases, in particular HCV RNA-dependent RNA polymerase (RdRp), rather than cellular nucleic acid polymerases. Compounds of formula V have been discovered to be effective against both wild-type and S282T mutant strains of the HCV virus. Thus, the compounds of formula V are useful for the treatment of Flaviviridae infections in humans and other animals.

In one embodiment, compounds of formula V are provided

or pharmaceutically acceptable salts thereof;

where R ¹ represents Ci-C^-alkyl;

R ^a represents -C(=O)R ¹¹ ;

R ⁷ represents

Y represents O;

W ¹ and W2 each represent, independently, a group of formula IVa

^Rx

formula IVa where each Y ¹ represents O;

each Y ² represents O or NR;

M2 represents 0;

each R ^x represents a group of formula IVb

- 2 046452 where each M1a, M1c and Mid represents 0;

M12c represents 0 or 1;

each R ^y independently represents H, -C(=Y1)OR, -OC(=Y1)R, _C1-8 alkyl, C ₆ -C ₂₀ -aryl, or C7-C ₂ o-arylalkyl; where each C ₆ -C ₂₀ -aryl in R ^y is unsubstituted or substituted by 1-3 R ²⁰ groups; and each R is, independently, H, C ₁ -C ₈ -alkyl or C ₃ -C ₂₀ -carbocyclyl;

wherein each C _1-8 alkyl in R is optionally substituted with one or more hydroxy;

R ⁸ represents NH ₂ ;

each R ¹¹ represents, independently, H or C1-C ₈ -alkyl; and each R ²⁰ is, independently, halogen.

In another embodiment, pharmaceutical compositions are provided containing an effective amount of a compound of Formula V, or a pharmaceutically acceptable salt thereof, in combination with a pharmaceutically acceptable diluent or carrier.

Detailed Description of Exemplary Embodiments

Reference will now be made in detail to certain embodiments of the invention, examples of which are shown in the accompanying specifications, structures and claims. While the invention will be described in connection with the listed embodiments, it should be understood that they are not intended to limit the invention in these embodiments. On the contrary, the invention is intended to cover all alternatives, modifications and equivalents that may be included within the scope of the present invention.

In another aspect, the compounds of formula I are represented by formula II

R ⁸

Formula II or a pharmaceutically acceptable salt thereof;

where R ¹ represents (C1- _{C8 )} alkyl, ( _C4 - _C8 )carbocyclylalkyl, (C1- _C8 )substituted alkyl, ( _C2C8 )alkenyl, ( _C2 - _C8 )substituted alkenyl, ( C ₂ -C ₈ )alkynyl, (C ₂ -C ₈ )substituted alkynyl or aryl (C1C ₈ )alkyl;

each R ³ , R ⁴ , R ⁵ are, independently, H, OR ^a , N(R ^a )2, N3, CN, NO2, S(O)nR ^a , halogen, (C1-C8)alkyl, (C ₄ -C ₈ )carbocyclylalkyl, (C1-C ₈ )substituted alkyl, (C ₂ -C ₈ )alkenyl, (C ₂ -C ₈ ) substituted alkenyl, (C ₂ -C ₈ )alkynyl, (C ₂ -C ₈ ) substituted alkynyl or aryl (C1- _C8 )alkyl;

or any two of R ³ , R ⁴ and R ⁵ on adjacent carbon atoms, taken together, are -O(CO)O- or, taken together with the carbon atoms of the ring to which they are attached, form a double bond;

R ⁶ represents H, OR ^a , N(R ^a )2, N3, CN, NO2, S(O)nR ^a , -C(=O)R ¹¹ , -C(=O)OR ¹¹ , -C( =O)NR11R ¹² , -C(=O)SR ¹¹ , -S(O)R ¹¹ , -S(O)2R ¹¹ , -S(O)(OR ¹¹ ), -S(O)2(OR ¹¹ ), -SO2NR ¹¹ R ¹² , halogen, (^^alkyl, (C4C8)carbocyclylalkyl, (^-^substituted alkyl, (^-^alkenyl, (C ₂ -C ₈ ) substituted alkenyl, (C ₂ ^alkynyl, ( C ₂ -C ₈ ) substituted alkynyl or aryl (^-O^alkyl;

each n, independently, is 0, 1 or 2;

each R ^a , independently, represents H, (^-^alkyl, (^-^alkenyl, (^-^alkynyl, aryl (b^alkyl, ^^carbocyclylalkyl, -C(=O)R ¹¹ , -C( =O)OR11, -C(=O)NR ¹¹ R ¹² , -C(=O)SR ¹¹ , -S(O)R11, -S(O) ₂ R ¹¹ , -S(O)(OR ¹¹ ) , -S(O)2(OR ¹¹ ) or -SO2NR11R ¹² ;

R ⁷ represents H, -C(=O)R ¹¹ , -C(=O)OR ¹¹ , -C(=O)NR ¹¹ R ¹² , -C(=O)SR ¹¹ , -S(O)R ¹¹ , -S(O)2R ¹¹ , -S(O)(OR11), -S(O)2(OR ¹¹ ), -SO2NR11R ¹² or

each Y or Y ¹ is, independently, O, S, NR, ⁺ N(O)(R), N(OR), ⁺ N(O)(OR) or N-NR ₂ ;

W ¹ and W ² taken together are -Y ³ (C(R ^y ) ₂ ) ₃ Y ³ -; or one of W ¹ or W ² , together with either R ³ or R ⁴ , is -Y ³ - and the other W ¹ or W ² is of formula Ia; or W ¹ and W ² , each independently, is a group of formula Ia

- 3 046452

Formula la where each Y ² represents, independently, a bond, O, CR2, NR, +N(O)(R), N(OR), ⁺ N(O)(OR), NNR2, S, SS, S( O) or S(O)2;

each Y ³ is independently O, S or NR;

M2 is 0, 1 or 2;

each R ^x represents, independently, R ^y or the formula

where each M1a, M1c and M1d is, independently, 0 or 1;

M12c is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12;

each R ^y represents, independently, H, F, Cl, Br, I, OH, R, -C(=Y1)R, -C(=Y ¹ )OR, -C(=Y ¹ )N(R) 2, -N(R)2, -+N(R)3, -SR, -S(O)R, -S(O)2R, -S(O)(OR), -S(O)2( OR), -OC(=Y1)R, -OC(=Y1)OR, -OC(=Y ¹ )(N(R)2), -SC(=Y1)R, -SC(=Y1)OR, -SC(=Y ¹ )(N(R) ₂ ), -N(R)C(=Y1)R, -N(R)C(=Y1)OR,

-N(R)C(=Y1)N(R) ₂ , -SO ₂ NR ₂ , -CN, -N ₃ , -NO ₂ , -OR or W ³ ; or, taken together, two ^Ry on the same carbon atoms form carbocyclic rings of from 3 to 7 carbon atoms;

each R, independently, represents H, (^-^alkyl, (^-^substituted alkyl, (C ₂ ^)alkenyl, (C ₂ -C ₈ ) substituted alkenyl, (^-^alkynyl, (C ₂ -C ₈ ) substituted alkynyl, C ₆ -C ₂₀ -aryl, C ₆ -C ₂₀ substituted aryl, C ₂ -C ₂₀ -heterocyclyl, C ₂ -C ₂₀ substituted heterocyclyl, arylalkyl or substituted arylalkyl;

W ³ is W ⁴ or W ⁵ ; W ⁴ is R, -C(Y1)R ^y , -C(Y1)W ⁵ , -SO2R ^y or -SO2W ⁵ ; and W ⁵ is a carbocycle or heterocycle, wherein W ⁵ is independently substituted with 0 to 3 R ^y groups;

each X ¹ or X ² is independently CR ¹⁰ or N;

each R ⁸ is a halogen, NR11R ¹² , N(R ¹¹ )OR ¹¹ , NR ¹¹ NR ¹¹ R ¹² , N3, NO, NO2, CHO, CN, -CH(=NR ¹¹ ), -CH=NNHR ¹¹ , - CH=N(OR ¹¹ ), -CH(OR ¹¹ )2, -C(=O)NR ¹¹ R ¹² , -C(=S)NR ¹¹ R ¹² , -C(=O)OR ¹¹ , (C1^ )alkyl, (^-^alkenyl, (^-^alkynyl, (C4-C ₈ )carbocyclylalkyl, optionally substituted aryl, optionally substituted heteroaryl, -C^OX^^^kil, -S(O)N(C ₁ - C ₈ )alkyl, aryl(C ₁ -C ₈ )alkyl, OR ¹¹ or SR ¹¹ ;

R ⁹ and R ¹⁰ are each independently H, halogen, NR11R ¹² , N(R ¹¹ )OR ¹¹ , NR ¹¹ NR ¹¹ R ¹² , N ₃ , NO, NO2, CHO, CN, -CH(=NR ¹¹ ), -CH=NHNR ¹¹ , -CH=N(OR ¹¹ ), -CH(OR ¹¹ )2, -C(=O)NR ¹¹ R ¹² , -C(=S)NR ¹¹ R ¹² , -C( =O)OR ¹¹ , R ¹¹ , OR ¹¹ or SR ¹¹ ;

each R ¹¹ or R ¹² , independently, represents H, (^-^alkyl, (^-^alkenyl, ( _C2 ^)alkynyl, (C4-C8)carbocyclylalkyl, optionally substituted aryl, optionally substituted heteroaryl, -C^ OX^-C^^kil, -SCOVCkQ^kuh or aryl (^-^alkyl; or R ¹¹ and R ¹² , taken together with the nitrogen to which they are both attached, form from 3 to 7-membered heterocyclic rings, where any one carbon atom of said heterocyclic ring may optionally be replaced by -O-, -S- or -NR ^a -;

where each (^-^alkyl, (^-^alkenyl, (^-^alkynyl or aryl (C ₁ -C ₈ ))alkyl of each R ¹ , R ³ , R ⁴ , R ⁵ , R ⁶ , R ¹¹ or R ¹² is, independently, optionally substituted with one or more halogens, hydroxy, CN, N3, N(R ^a )2 or OR ^a ; and wherein one or more of the non-terminal carbon atoms of each (^-^alkyl, optionally, may be replaced by - O- , -S- or -NR ^a -.

In one embodiment of the invention of Formula II, R ¹ is (C1-C8)alkyl, (^-^alkenyl, or (^-^alkynyl). In another aspect of this embodiment, R ¹ is (C1^)alkyl. In another aspect thereof embodiments, R ¹ is methyl, CH2F, or ethynyl. In another aspect of this embodiment, R ¹ is (C1-C 8 )alkyl, and R ⁶ is H. In another aspect of this embodiment, R ¹ is (C1-C ₈ )alkyl. R ¹ is (C1-C8)alkyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ¹ is (C1

- 4 046452

C8)alkyl, and R ⁶ is CN, OH or CH3.

In one embodiment of Formula II, R3 is H, OR ^a , N(R ^a )2, N3, CN, SR ^a , halogen, (^-^alkyl, (^-^alkenyl, or (^-^alkynyl). B In one aspect of this embodiment, R ³ is H. In another aspect of this embodiment, R ³ is H, and R 1 is (C1-C8)alkyl, (C2-C8)alkenyl, or ( _C2 -C8)alkynyl. In another aspect thereof. embodiments ^R3 is H and R1 is (C1-C8)alkyl. In another aspect of this embodiment, ^R3 is H and R1 is methyl, CH2F or ethynyl. In another aspect of this embodiment, ^R3 is H and R1 is H. is methyl. In another aspect of this embodiment, R ³ is H, R 1 is (C ₁ C8)alkyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ³ is H, R ¹ is methyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ³ is H, R ¹ is (C1 -^alkyl, and R ⁶ is CN, OH or CH3 . In another aspect of this embodiment, R ³ is H, R ¹ is methyl, and R ⁶ is H.

In one embodiment of Formula II, R ⁴ is H, OR ^a , N(R ^a )2, N3, CN, SR ^a , halogen, (^-^alkyl, (^-^alkenyl, or (^-^alkynyl). B In another aspect of this embodiment, R ⁴ is H or OR ^a . In another aspect of this embodiment, R ⁴ is OR ^a . In another aspect of this embodiment, R ⁴ is OR ^a , and R ¹ is (^-^alkyl, (^-^alkenyl). or (^-^alkynyl. In another aspect of this embodiment, R ⁴ is OR ^a , and R ¹ is (C1-C ₈ )alkyl, (^-^alkenyl, or (^-^alkynyl. In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is (C1-C8)alkyl. In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OR ^a , R. ¹ is (C1^alkyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is (^-^alkyl, and R ⁶ is CN, OH or CH3. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and R ⁶ is CN, OH or CH3. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and R ⁶ is H. In another aspect of this embodiment, R ⁴ is OH, and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OH, R ¹ is (^-^alkyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OH, R ¹ is (^-^alkyl, and R ⁶ is CN, OH or CH3. In another in an aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and R ⁶ is CN, OH, or CH3. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and R ⁶ is H.

In one embodiment of Formula II, R ⁵ is H, OR ^a , N(R ^a )2, N3, CN, SR ^a , halogen, (^^8)ηλκηλ, (^-^alkenyl or (^-^alkynyl. In another aspect of this embodiment ^, R ⁴ is H or OR ^a . In another aspect of this embodiment ^, R ⁴ is OR ^a , and R ¹ is (^-^)ηλκηλ. (^-^alkenyl or (^-^alkynyl. In another aspect of this embodiment, R ⁴ is OR ^a , and R ¹ is (^-^alkyl, (^-^alkenyl, or (^-^alkynyl. In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is (^-^)ηλκηλ. In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OR. ^a , R ¹ is (C1 -^alkyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is (^-^alkyl, and R ⁶ is CN, OH or CH3. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and R ⁶ is CN, OH or CH3. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and R ⁶ is H. In another aspect of this embodiment, R ⁴ is OH, and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OH, R ¹ is (C ₁ -C ₈ )alkyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OH, R ¹ is (^-^alkyl, and R ⁶ is CN, OH or CH3. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and R ⁶ is CN, OH, or CH3. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and R. ⁶ is H. In another aspect of this embodiment, R ⁵ is N3.

In another aspect of the embodiment of Formula II, R ⁵ is H. In another aspect of this embodiment, R ⁴ is H or OR ^a . In another aspect of this embodiment, R ⁴ is OR ^a . In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is (C ₁ ^alkyl, (^-^alkenyl or (^-^alkynyl). In another aspect of this embodiment, R ⁴ is

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OR ^a and R ¹ is (C1-C8)alkyl, (C2-C8)alkenyl or (C2-C8)alkynyl. In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is (C1-C ₈ )alkyl. In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is (C1-C8)alkyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OR ^a . R ¹ is methyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is (C1-C8)alkyl, and R ⁶ is CN , OH or CH3. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and R ⁶ is CN, OH or CH3. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and R ⁶ is H. In another aspect of this embodiment, R ⁴ is OH, and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OH, R ¹ is (C1 -C8)alkyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OH, R ¹ is (C1-C8)alkyl, and R ⁶ is CN, OH, or CH3. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and R ⁶ is CN, OH or CH ₃ . In another aspect of this embodiment, ^R4 is OH, ^R1 is methyl, and ^R6 is H.

In another embodiment of Formula II, R ⁶ is H, CN, OR ^a or CH3. In another aspect of this embodiment, R ⁶ is H. In another aspect of this embodiment, R ⁶ is CN. In another aspect of this embodiment, R ⁶ is OR ^a . In another aspect of this embodiment, R ⁶ is OH. In another aspect of this embodiment, R ⁶ is CH3. In another aspect of this embodiment, R ⁴ represents H or OR ^a . In another aspect of this embodiment, R ⁴ is OR ^a . In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is (C1-C8)alkyl, (C2-C8)alkenyl or (C2-C8)alkynyl. In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is (C1-C8)alkyl, (C2-C8)alkenyl or ( _C2 -C8)alkynyl. In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is (C1-C8)alkyl. In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is (C1 -C8)alkyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OR ^a . R ¹ is methyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is (C1-C8)alkyl, and R ⁶ is CN , OH or CH3. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and R ⁶ is CN, OH or CH3. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and R ⁶ is H. In another aspect of this embodiment, R ⁴ is OH, and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OH, R ¹ is (C1-C8)alkyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OH, R ¹ is (C1-C8)alkyl, and R ⁶ is CN, OH, or CH3. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and R ⁶ is CN, OH or CH3. In another aspect of this embodiment, ^R4 is OH, ^R1 is methyl, and ^R6 is H.

In another embodiment of Formula II, R ⁶ is CN, OR ^a or CH3. In another aspect of this embodiment, R ⁶ is CN. In another aspect of this embodiment, R ⁶ is OR ^a . In another aspect of this embodiment, R ⁶ is OH. In another aspect of this embodiment, R ⁶ is CH3. In another aspect of this embodiment, R ⁴ represents H or OR ^a . In another aspect of this embodiment, R ⁴ is OR ^a . In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is (C1-C8)alkyl, (C2-C8)alkenyl or (C20)alkynyl. In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is (G-C8)onkug, (C ₂ -C8)alkenyl or (0-D)alkynyl. In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is (C1-C8)alkyl. In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is (C1 -C8)alkyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OR ^a . R ¹ is methyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is (C1-C8)alkyl, and R ⁶ is CN , OH or CH3. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and R ⁶ is CN, OH or CH3. In another aspect of this embodiment, R ⁴ is OH and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OH, R ¹ is (C1C ₈ )alkyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and at least one of X ¹ or X ² before

- 6 046452 is N. In another aspect of this embodiment, R ⁴ is OH, R ¹ is (C ₁ -C ₈ )alkyl, and R ⁶ is CN, OH or CH3. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and R ⁶ is CN, OH or CH3.

In one embodiment of Formula II, R ⁷ is H, -C(=O)R ¹¹ , -C(=O)OR ¹¹ , -C(=O)SR ¹¹ or

In an aspect of this embodiment, R ⁷ is H. In another aspect of this embodiment, R ⁷ is -C(=O)R ¹¹ . In another aspect of this embodiment, R ⁷ is -C(=O)R ¹¹ , where R ¹¹ is (C1-C ₈ )alkyl.

In another aspect of this embodiment, R ⁷ is

In another aspect of this embodiment, R ⁶ is H. In another aspect of this embodiment, R ⁶ is CN. In another aspect of this embodiment, R ⁶ is OR ^a . In another aspect of this embodiment, R ⁶ is OH. In another aspect of this embodiment, R ⁶ is CH3. In another aspect of this embodiment, R ⁴ represents H or OR ^a . In another aspect of this embodiment, R ⁴ is OR ^a . In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is (C1-C8)alkyl, (C2-C8)alkenyl or (C2-C8)alkynyl. In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is (C1-C8)alkyl. In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is (C1-C8)alkyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OR ^a . R ¹ is methyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is (C1-C8)alkyl, and R ⁶ is CN , OH or CH3. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and R ⁶ is CN, OH or CH3. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and R ⁶ is H. In another aspect of this embodiment, R ⁴ is OH, and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OH, R ¹ is (C1-C8)alkyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OH, R ¹ is (C1-C8)alkyl, and R ⁶ is CN, OH, or CH3. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and R ⁶ is CN, OH or CH3. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and R ⁶ is H.

In one embodiment of Formula II, X ¹ is N or CR ¹⁰ . In another aspect of this embodiment, X ¹ is N. In another aspect of this embodiment, X ¹ is CR ¹⁰ . In another aspect of this embodiment, X ² represents CH. In another aspect of this embodiment, X ¹ is N and X ² is CH. In another aspect of this embodiment, X ¹ is CR ¹⁰ and X ² is CH. In another aspect of this embodiment, R ⁶ is H. In another aspect of this embodiment, R ⁶ is CN. In another aspect of this embodiment, R ⁶ is OR ^a . In another aspect of this embodiment, R ⁶ is OH. In another aspect of this embodiment, R ⁶ is CH3. In another aspect of this embodiment, R ⁴ represents H or OR ^a . In another aspect of this embodiment, R ⁴ is OR ^a . In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is (C1-C8)alkyl, (C2C8)alkenyl or (C2-C8)alkynyl. In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is (C1-C8)alkyl. In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is (C1-C8)alkyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OR ^a . R ¹ is methyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is (C1-C8)alkyl, and R ⁶ is CN , OH or CH3. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and R ⁶ is CN, OH or CH3. In another aspect of this embodiment, R ⁴ is OH and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OH, R ¹ is (C1-C ₈ )alkyl, and

- 7 046452 at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and at least one of X ¹ or X ² is N. In another In an aspect of this embodiment, R ⁴ is OH, R ¹ is (C1-C ₈ )alkyl, and R ⁶ is CN, OH or CH3. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and R ⁶ is CN, OH or CH3. In another aspect of this embodiment, ^R4 is OH, ^R1 is methyl, and ^R6 is H.

In another embodiment of Formula II, each R ⁸ is, independently, halogen, NR11R ¹² , N(R ¹¹ )OR ¹¹ , NR ¹¹ NR ¹¹ R ¹² , OR ¹¹ or SR ¹¹ . In another aspect of this embodiment, R ¹ is methyl, CH2F or ethynyl. In another aspect of this embodiment, R ¹ is methyl. In another aspect of this embodiment, R ⁹ is H, halogen or NR ¹¹ R ¹² . In another aspect of this embodiment, R ⁹ is H, halogen, or NR ¹¹ R ¹² , and R ¹ is methyl, CH2F or ethynyl. In another aspect of this embodiment, R ⁹ is H, halogen or NR ⁿ R ¹² , and R ¹ is methyl. In another aspect of this embodiment, R ⁸ is NH2 and R ⁹ is H or halogen. In another aspect of this embodiment, R ⁸ is NH2 and R ⁹ is H or halogen, and R ¹ is methyl, CH2F or ethynyl. In another aspect of this embodiment, R ⁸ is NH2, and R ⁹ is H or halogen, and R ¹ is methyl. In another aspect of this embodiment, R ⁸ and R ⁹ are each NH2. In another aspect of this embodiment, R ⁸ and R ⁹ are each NH2, and R ¹ is methyl. In another aspect of this embodiment, R ⁸ and R ⁹ are each NH2, and R ¹ is methyl, CH2F or ethynyl. In another aspect of this embodiment, R ⁸ is OH and R ⁹ is NH2. In another aspect of this embodiment, R ⁸ is OH, R ⁹ is NH2, and R ¹ is methyl. In another aspect of this embodiment, R ⁸ is OH, R ⁹ is NH2, and R ¹ is methyl, CH2F or ethynyl. In another aspect of this embodiment, R ⁶ is H. In another aspect of this embodiment, R ⁶ is CN. In another aspect of this embodiment, R ⁶ is OR ^a . In another aspect of this embodiment, R ⁶ is OH. In another aspect of this embodiment, R ⁶ is CH3. In another aspect of this embodiment, R ⁴ represents H or OR ^a . In another aspect of this embodiment, R ⁴ is OR ^a . In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is (C1-C8)alkyl, (C2-C8)alkenyl or (C2-C8)alkynyl. In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is (C1-C8)alkyl. In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is (C1-C8)alkyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OR ^a . R ¹ is methyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is (C1-C8)alkyl, and R ⁶ is CN , OH or CH3. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and R ⁶ is CN, OH or CH3. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and R ⁶ is H. In another aspect of this embodiment, R ⁴ is OH, and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OH, R ¹ is (C1-C8)alkyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OH, R ¹ is (C1-C8)alkyl, and R ⁶ is CN, OH, or CH3. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and R ⁶ is CN, OH or CH3. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and R ⁶ is H.

In another embodiment of Formula II, each R ¹⁰ is, independently, H, halogen, CN, or optionally substituted heteroaryl. In another aspect of this embodiment, R ¹ is methyl. In another aspect of this embodiment, R ¹ is methyl, CH2F or ethynyl. In another aspect of this embodiment, R ⁹ is H, halogen or NR11R ¹² . In another aspect of this embodiment, R ⁹ is H, halogen or NR11R ¹² , and R ¹ is methyl. In another aspect of this embodiment, R ⁹ is H, halogen or NR ⁿ R ¹² , and R ¹ is methyl, CH2F or ethynyl. In another aspect of this embodiment, R ⁸ is NH2 and R ⁹ is H or halogen. In another aspect of this embodiment, R ⁸ is NH2, and R ⁹ is H or halogen, and R ¹ is methyl. In another aspect of this embodiment, R ⁸ is NH2 and R ⁹ is H or halogen, and R ¹ is methyl, CH2F or ethynyl. In another aspect of this embodiment, R ⁸ and R ⁹ are each NH2. In another aspect of this embodiment, R ⁸ and R ⁹ are each NH2, and R ¹ is methyl. In another aspect of this embodiment, R ⁸ and R ⁹ are each NH2, and R ¹ is methyl, CH2F or ethynyl. In another aspect of this embodiment, R ⁸ is OH and R ⁹ is NH2. In another aspect of this embodiment, R ⁸ is OH, R ⁹ is NH2, and R ¹ is methyl. In another aspect of this embodiment, R ⁸ is OH, R ⁹ is NH ₂ , and R ¹ is methyl, CH2F or ethynyl. In another aspect of this embodiment, R ⁶ is H. In another aspect

- 8 046452 of this embodiment R ⁶ represents CN. In another aspect of this embodiment, R ⁶ is OR ^a . In another aspect of this embodiment, R ⁶ is OH. In another aspect of this embodiment, R ⁶ is CH3. In another aspect of this embodiment, R ⁴ is H or OR ^a In another aspect of this embodiment, R ⁴ is OR ^a In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is (C1-C8)alkyl, (C2 -C8)alkenyl or (C2-C8)alkenyl. In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is (C1-C ₈ )alkyl. In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is (C1-C8)alkyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OR ^a . R ¹ is methyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is (C1-C ₈ )alkyl, and R ⁶ is CN, OH or CH3. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and R ⁶ is CN, OH or CH3. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and R ⁶ is H. In another aspect of this embodiment, R ⁴ is OH, and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OH, R ¹ is (C1-C8)alkyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OH, R ¹ is (C1-C ₈ )alkyl, and R ⁶ is CN, OH or CH3. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and R ⁶ is CN, OH or CH3. In another aspect of this embodiment, ^R4 is OH, ^R1 is methyl, and ^R6 is H.

In another embodiment, the compounds of formula I or formula II are represented by formula III

R ⁸

^R4F

Formula III or a pharmaceutically acceptable salt thereof;

where R ¹ represents CH3, CH2F or ethynyl, and all other variables are defined as for formula I.

In one embodiment of Formula III, R ⁴ is H, OR ^a , N(R ^a )2, N3, CN, SR ^a , halogen, (^-^alkyl, (^-^alkenyl, or (^-^alkynyl). B in another aspect of this embodiment ^, R ⁴ is H or OR ^a . In another aspect of this embodiment ^{, R 4 is} OR ^a , and R ¹ is CH3, CH2F, or ethynyl. in an aspect of this embodiment, R ⁴ is OR ^a and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and R ⁶ is CN, OH, or CH3. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and R ^6. is H. In another aspect of this embodiment, R ⁴ is OH, and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and at least one of X ¹ or X ^2. represents N. In another aspect of this embodiment, R ⁴ represents OH, R ¹ represents methyl, and R ⁶ represents CN, OH or CH ₃ . In another aspect of this embodiment, ^R4 is OH, ^R1 is methyl, and ^R6 is H.

In another embodiment of Formula III, R ⁶ is H, CN, OR ^a or CH3. In another aspect of this embodiment, R ⁶ is H. In another aspect of this embodiment, R ⁶ is CN. In another aspect of this embodiment, R ⁶ is OR ^a . In another aspect of this embodiment, R ⁶ is OH. In another aspect of this embodiment, R ⁶ is CH3. In another aspect of this embodiment, R ⁴ represents H or OR ^a . In another aspect of this embodiment, R ⁴ is OR ^a . In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl , and R ⁶ represents CN, OH or

- 9 046452

_CH3 . In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and R ⁶ is H. In another aspect of this embodiment, R ⁴ is OH, and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and R ⁶ is CN, OH or CH3. In another aspect of this embodiment, ^R4 is OH, ^R1 is methyl, and ^R6 is H.

In another embodiment of Formula III, R ⁶ is CN, OR ^a or CH3. In another aspect of this embodiment, R ⁶ is CN. In another aspect of this embodiment, R ⁶ is OR ^a . In another aspect of this embodiment, R ⁶ is OH. In another aspect of this embodiment, R ⁶ is CH3. In another aspect of this embodiment, R ⁴ represents H or OR ^a . In another aspect of this embodiment, R ⁴ is OR ^a . In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl and R ⁶ is CN, OH or CH3. In another aspect of this embodiment, R ⁴ is OH and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and R ⁶ is CN, OH or CH ₃ .

In another embodiment of Formula III, R ⁷ is H, -C(=O)R ¹¹ , -C(=O)OR ¹¹ , -C(=O)SR ¹¹ or

In an aspect of this embodiment, R ⁷ is H. In another aspect of this embodiment, R ⁷ is -C(=O)R ⁿ . In another aspect of this embodiment, R ⁷ is -C(=O)R ⁿ , where R ¹¹ is (^-^alkyl.

In another aspect of this embodiment, R ⁷ is

In another aspect of this embodiment, R ⁶ is H. In another aspect of this embodiment, R ⁶ is CN. In another aspect of this embodiment, R ⁶ is OR ^a . In another aspect of this embodiment, R ⁶ is OH. In another aspect of this embodiment, R ⁶ is CH3. In another aspect of this embodiment, R ⁴ is H or OR ^a In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl and R ⁶ is CN, OH or CH3. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and R ⁶ is H. In another aspect of this embodiment, R ⁴ is OH, and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and R ⁶ is CN, OH or CH3. In another aspect of this embodiment, ^R4 is OH, ^R1 is methyl, and ^R6 is H.

In one embodiment of Formula III, X ¹ is N or CR ^B*10 . In another aspect of this embodiment, X ¹ is N. In another aspect of this embodiment, X ¹ is CR ¹⁰ . In another aspect of this embodiment, X ² represents CH. In another aspect of this embodiment, X ¹ is N and X ² is CH. In another aspect of this embodiment, X ¹ is CR ¹⁰ and X ² is CH. In another aspect of this embodiment, R ⁶ is H. In another aspect of this embodiment, R ⁶ is CN. In another aspect of this embodiment, R ⁶ is OR ^a . In another aspect of this embodiment, R ⁶ is OH. In another aspect of this embodiment, R ⁶ is CH ₃ . In another aspect of this embodiment, R ⁴ represents H or OR ^a . In another aspect of this embodiment, R ⁴ is OR ^a . In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and at least one of

- 10 046452

X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and R ⁶ is CN, OH or CH3. In another aspect of this embodiment, R ⁴ is OH and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and R ⁶ is CN, OH or CH ₃ . In another aspect of this embodiment, ^R4 is OH, ^R1 is methyl, and ^R6 is H.

In another embodiment of Formula III, each R ⁸ is, independently, halogen, NR ¹¹ R ¹² , N(R ¹¹ )OR ¹¹ , NR ¹¹ NR ¹¹ R ¹² , OR ¹¹ or SR ¹¹ . In another aspect of this embodiment, R ¹ is methyl, CH2F or ethynyl. In another aspect of this embodiment, R ¹ is methyl. In another aspect of this embodiment, R ⁹ is H, halogen or NR ¹¹ R ¹² . In another aspect of this embodiment, R ⁹ is H, halogen or NR ⁿ R ¹² , and R ¹ is methyl, CH2F or ethynyl. In another aspect of this embodiment, R ⁹ is H, halogen or NR ⁿ R ¹² , and R ¹ is methyl. In another aspect of this embodiment, R ⁸ is NH2 and R ⁹ is H or halogen. In another aspect of this embodiment, R ⁸ is NH2 and R ⁹ is H or halogen, and R ¹ is methyl, CH2F or ethynyl. In another aspect of this embodiment, R ⁸ is NH2, and R ⁹ is H or halogen, and R ¹ is methyl. In another aspect of this embodiment, R ⁸ and R ⁹ are each NH2. In another aspect of this embodiment, R ⁸ and R ⁹ are each NH2, and R ¹ is methyl, CH2F or ethynyl. In another aspect of this embodiment, R ⁸ and R ⁹ are each NH2, and R ¹ is methyl. In another aspect of this embodiment, R ⁸ is OH and R ⁹ is NH2. In another aspect of this embodiment, R ⁸ is OH, R ⁹ is NH2, and R ¹ is methyl, CH2F or ethynyl. In another aspect of this embodiment, R ⁸ is OH, R ⁹ is NH2, and R ¹ is methyl. In another aspect of this embodiment, R ⁶ is H. In another aspect of this embodiment, R ⁶ is CN. In another aspect of this embodiment, R ⁶ is OR ^a . In another aspect of this embodiment, R ⁶ is OH. In another aspect of this embodiment, R ⁶ is CH3. In another aspect of this embodiment, R ⁴ represents H or OR ^a . In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl and R ⁶ is CN, OH or CH3. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and R ⁶ is H. In another aspect of this embodiment, R ⁴ is OH, and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and R ⁶ is CN, OH or CH3. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and R ⁶ is H.

In another embodiment of Formula III, each R ¹⁰ is, independently, H, halogen, CN, or optionally substituted heteroaryl. In another aspect of this embodiment, R ¹ is methyl. In another aspect of this embodiment, R ⁹ is H, halogen or NR ⁿ R ¹² . In another aspect of this embodiment, R ⁹ is H, halogen or NR ¹¹ R ¹² , and R ¹ is methyl. In another aspect of this embodiment, R ⁸ is NH2 and R ⁹ is H or halogen. In another aspect of this embodiment, R ⁸ is NH2, and R ⁹ is H or halogen, and R ¹ is methyl. In another aspect of this embodiment, R ⁸ and R ⁹ are each NH2. In another aspect of this embodiment, R ⁸ and R ⁹ are each NH2, and R ¹ is methyl. In another aspect of this embodiment, R ⁸ is OH and R ⁹ is NH2. In another aspect of this embodiment, R ⁸ is OH, R ⁹ is NH2, and R ¹ is methyl. In another aspect of this embodiment, R ⁶ is H. In another aspect of this embodiment, R ⁶ is CN. In another aspect of this embodiment, R ⁶ is OR ^a . In another aspect of this embodiment, R ⁶ is OH. In another aspect of this embodiment, R ⁶ is CH3. In another aspect of this embodiment, R ⁴ represents H or OR ^a . In another aspect of this embodiment, R ⁴ is OR ^a and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl and R ⁶ is CN, OH or CH3. In another aspect of this embodiment, R ⁴ is OR ^a , R ¹ is methyl, and R ⁶ is H. In another aspect of this embodiment, R ⁴ is OH, and R ¹ is methyl. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and at least one of X ¹ or X ² is N. In another aspect of this embodiment, R ⁴ is OH, R ¹ is methyl, and R ⁶ is CN, OH or CH3. In another aspect of this embodiment, ^R4 is OH, ^R1 is methyl, and ^R6 is H.

In another embodiment, compounds of formula IV are provided

- 11 046452

or pharmaceutically acceptable salts thereof;

where R ¹ represents (Cr3Dalkyl, ( _C4 - _C8 )carbocyclylalkyl, (CrC^substituted alkyl, ( _C2C8 )alkenyl, ( _C2 - _C8 )substituted alkenyl, (CVC^ _llikuiuj , ( _C2 - C ₈ )substituted alkynyl or aryl(C1C ₈ )alkyl;

R ² represents halogen;

R ³ , R ⁴ and R ⁵ are each independently H, halogen, OR ^a , N(R ^a )2, N3, CN, NO2, S(O)nR ^a , (C1-C8)alkyl, (C ₄ -C ₈ ) carbocyclyl alkyl, (C 1 - C ₈ ) substituted alkyl, (C ₂ - C ₈ ) alkenyl, (C ₂ - C ₈ ) substituted alkenyl, (C ₂ - C ₈ ) alkynyl, (C ₂ - _{C 8} )substituted alkynyl or aryl(C1- _C8 )alkyl;

or any two of R ³ , R ⁴ or R ⁵ on adjacent carbon atoms, taken together, are -O(CO)O, or, taken together with the carbon atoms of the ring to which they are attached, form a double bond;

each n is, independently, 0, 1 or 2;

each R ^a represents, independently, H, (C1- _C8 )alkyl, ( _C2 - _C8 )alkenyl, ( _C2 - _C8 )kynyl, aryl(C1- _C8 )alkyl, (C4- _C8 )carbocyclylalkyl, -C(=O)R ⁱ¹ , -C(=O)OR ⁱ¹ , -C(=O)NR ¹¹ R ¹² , -C(=O)SR ⁱ¹ , -S(O)R ¹¹ , - S(O)2R ¹¹ , -S(O)(OR ¹¹ ), -S(O)2(OR ¹¹ ) or -SO2NR ¹¹ R ¹² ;

R ⁷ represents H, -C(=O)R ¹¹ , -C(=O)OR ⁱ¹ , -C(=O)NR ⁱ¹ R ¹² , -C(=O)SR ¹¹ , -S(O)R ⁱ¹ , -S(O)2R ¹¹ , -S(O)(OR ⁱ¹ ), -S(O)2(OR ⁱ¹ ), -SO2NR ¹ⁱ R ⁱ² , or

Y is O, S, NR, +N(O)(R), N(OR), ⁺ N(O)(OR) or N-NR2;

W ¹ and W ² taken together are -Y ³ (C(R ^y ) ₂ ) ₃ Y ³ -; or one of W ¹ or W ² , together with either R ³ or R ⁴ , is -Y ³ - and the other W ¹ or W ² is of formula Ia; or

W ¹ and W ² are, each independently, a group of formula IVa

^Rx

Formula IVa where each Y ¹ is, independently, O, S, NR, +N(O)(R), N(OR), ⁺ N(O)(OR) or N-NR2;

each Y ² represents, independently, a bond, O, CR2, NR, +N(O)(R), N(OR), ⁺ N(O)(OR), N-NR2, S, SS, S(O ) or S(O)2;

each Y ³ is independently O, S or NR;

M2 represents 0, 1 or 2;

each R ^x represents a group of formula IVb

- 12 046452

Formula IVb where each M1a, M1c and Mid is, independently, 0 or 1;

M12c represents 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12;

each R ^y represents, independently, H, F, Cl, Br, I, OH, -C(=Y1)R, -C(=Y1)R ¹³ , -C(=Y1)OR, -C(=Y1 )N(R)2, -N(R)2, - ⁺ N(R)3, -SR, -S(O)R, -S(O)2R, -S(O)2R ¹³ , -S( O)(OR), -S(O)2(OR), -OC(=Y ¹ )R, -OC(=Y ¹ )OR, -OC(=Y1)(N(R)2), -SC (=Y1)R, -SC(=Y1)OR, -SC(=Y1)(N(R) ₂ ), -N(R)C(=Y1)R, -N(R)C(=Y1) OR, -N(R)C(=Y1)N(R) ₂ , -SO2NR2, -CN, -N ₃ , -NO ₂ , -OR, (C _r C ₈ )^m, (^-^ alkenyl, (^-^alkynyl, C ₆ -C ₂₀ aryl, C ₃ -C ₂₀ -carbocyclyl, C ₂ -C ₂₀ -heterocyclyl, arylalkyl, heteroarylalkyl;

where each (^-^alkyl, (^-^alkenyl, (^-^alkynyl, C ₆ -C ₂₀ -aryl, C ₃ -C ₂₀ -carbocyclyl, C ₂ -C ₂₀ heterocyclyl, arylalkyl or heteroarylalkyl, optionally substituted 1-3 R^^ynm^;

or, taken together, two ^Ry on the same carbon atoms form carbocyclic rings of from 3 to 7 carbon atoms;

each R represents, independently, H, (^-^alkyl, (^-^alkenyl, (^-^alkynyl, C6-C20aryl, C3- _{C20 -} carbocyclyl, _C2 - _C20 -heterocyclyl, or arylalkyl);

each R ⁸ is halogen, NR11R ¹² , N(R ¹¹ )OR ¹¹ , NR ¹¹ NR ¹¹ R ¹² , N ₃ , NO, NO ₂ , OR ¹¹ or S(O)nR11;

each R ⁹ represents, independently, H, halogen, NR11R ¹² , N(R ¹¹ )OR ¹¹ , NR ^u NR11R ¹² , N3, NO, NO2, CHO, CN, -CH(=NR ¹¹ ), -CH=NHNR ¹¹ , -CH=N(OR ¹¹ ), -CH(OR ¹¹ ) ₂ , -C(=O)NR ¹¹ R ¹² , -C(=S)NR ¹¹ R ¹² , -C(=O)OR ¹¹ , R ¹¹ , OR ¹¹ or S(O)nR11;

each R ¹¹ or R ¹² is, independently, H, (^-^alkyl, (^-^alkenyl, ( _C2 ^)alkynyl, (C4- _C8 )carbocyclylalkyl, optionally substituted aryl, optionally substituted heteroaryl, -C (=O)(C ₁ -C ₈ )alkyl, -S(O) _n (C ₁ -C ₈ )alkyl or aryl^-^alkyl, or R ¹¹ and R ¹² , together with the nitrogen to which they are both attached; , form a 3-7 membered heterocycle, where any carbon atom of the specified heterocyclic ring can optionally be replaced by -O-, -S- or -NR ^b -;

each R ¹³ is, independently, a carbocycle or a heterocycle, optionally substituted with 1-3 R ²⁰ groups;

each R ²⁰ is, independently, halogen, CN, N ₃ , N(R) ₂ , OR, -SR, -S(O)R, -S(O) ₂ R, -S(O)(OR), -S(O) ₂ (OR), -C(=Y1)R, -C(=Y1)OR or C(=Y1)N(R) ₂ ;

where each (^-^alkyl, (^-^alkenyl, (^-^alkynyl or aryl^-^alkyl) of each R ¹ , R ³ , R ⁴ , R ⁵ , R ⁶ , R ¹¹ or R ¹² is, independently , optionally substituted with one or more halogens, hydroxy, CN, N3, N(R ^b ) 2 or OR ^b ; and wherein one or more non-terminal carbon atoms of each (^-^ alkyl may optionally be replaced by -O-, - S- or -NR ^b ;

each R ^b represents, independently, H, (^-^alkyl, (^-^alkenyl, (^-^alkynyl, aryl^^alkyl, ^^carbocyclylalkyl, -C(=O)R ²¹ , -C(= O)OR ²¹ , -C(=O)NR ²¹ R ²² , -C(=O)SR ²¹ , -S(O)R ²¹ , -S(O)2R ²¹ , -S(O)(OR ²¹ ) , -S(O)2(OR ²¹ ) or -SO2NR ²¹ R ²² ;

each R ²¹ or R ²² is, independently, H, (^-^alkyl, (^-^alkenyl, (C ₂ ^)alkynyl, (O^carbocyclylalkyl, -C(=O)(C ₁ -C ₈ ) alkyl, -S(O) _n (C ₁ -C ₈ )alkyl or aryl(C ₁ -C ₈ )alkyl;

with the optional provision that connections 1, 1d, 1e, 2, TP-1, A-1, 8 and 21 are excluded.

In another aspect of this embodiment, Y and Y ¹ is O. In another aspect of this embodiment, R ⁸ is halogen, NR11R ¹² , N(R ¹¹ )OR ¹¹ , NR ¹¹ NR ¹¹ R ¹² , OR ¹¹ or S(O)nR11 . In another aspect of this embodiment, R ⁹ is H, halogen, S(O)nR11 or NR11R ¹² . In another aspect of this embodiment, R ⁴ is ORa. In another aspect of this embodiment, R ¹ is CH3. In another aspect of this embodiment, R ² is F.

In another aspect of this embodiment, R ⁷ is

where Y is -O-; W ¹ represents formula Ia, and W ² , together with R ⁴ , represents -O-. In another embodiment, the compounds of formula IV are represented by formula V

- 13 046452

where R ¹ represents methyl or ethynyl, and R ⁴ represents OR ^a . In another aspect of this embodiment, R ⁷ is H or

In another aspect of this embodiment, the compounds of formula V are represented by the following structures

In another embodiment, compounds of formula VI are provided

- 14 046452

Formula VI or a pharmaceutically acceptable salt thereof;

where R ⁴ represents ORa;

each n is, independently, 0, 1 or 2;

each Ra, independently, represents H, (^-^alkyl, (^-^alkenyl, (^-^alkynyl, aryl (^^alkyl, (C4-C8)carbocyclylalkyl, -C(=O)R ¹¹ , - C(=O)OR ¹¹ , -C(=O)NR ¹¹ R ¹² , -C(=O)SR ¹¹ , -S(O)R ¹¹ , -S(O) ₂ R ¹¹ , -S(O) (OR ¹¹ ), -S(O)2(OR ¹¹ ) or -SO2NR ¹¹ R ¹² ;

R ⁷ represents H, -C(=O)R ¹¹ , -C(=O)OR ¹¹ , -C(=O)NR ¹¹ R ¹² , -C(=O)SR ¹¹ , -S(O)R ¹¹ , -S(O)2R ¹¹ , -S(O)(OR ¹¹ ), -S(O)2(OR ¹¹ ), -SO ₂ NR ¹¹ R ¹² or

Y is O, S, NR, +N(O)(R), N(OR), ⁺ N(O)(OR) or N-NR2;

W ¹ and W ² taken together are -Y ³ (C(R ^y )2) ₃ Y ³ -; or one of W ¹ or W ² , together with R ⁴ , is -Y ³ - and the other W ¹ or W ² is of formula Ia;

or

W ¹ and W ² each independently represent a group of the formula Via

Formula Via where each Y ¹ is, independently, O, S, NR, +N(O)(R), N(OR), ⁺ N(O)(OR) or N-NR2;

each Y ² represents, independently, a bond, O, CR2, NR, ⁺ N(O)(R), N(OR), ⁺ N(O)(OR), N-NR2, S, SS, S(O ) or S(O) ₂ ;

each Y ³ is independently O, S or NR;

M2 represents 0, 1 or 2;

each R ^x represents a group of formula VIb

where each M1a, M1c and M1d is, independently, 0 or 1;

M12c represents 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12;

each R ^y , independently, represents H, F, Cl, Br, I, OH, -C(=Y1)R, -C(=Y1)R ¹³ , -C(=Y1)OR, -C(=Y ¹ )N(R)2, -N(R)2, - ⁺ N(R)3, -SR, -S(O)R, -S(O)2R, -S(O)2R ¹³ , -S (O)(OR), -S(O)2(OR), -OC(=Y ¹ )R, -OC(=Y ¹ )OR, -OC(=Y ¹ )(N(R)2), -SC(=Y ¹ )R, -SC(=Y ¹ )OR, -SC(=Y ¹ )(N(R)2), -N(R)C(=Y ¹ )R, -N(R )C(=Y ¹ )OR, -N(R)C(=Y ¹ )N(R) ₂ , -SO2NR2, -CN, -N ₃ , -NO2, -OR, (C _r C ₈ ^m, (^-^alkenyl, (^-^alkynyl, C ₆ -C ₂₀

- 15 046452 aryl, C3-C ₂₀ -carbocyclyl, C ₂ -C ₂₀ -heterocyclyl, arylalkyl, heteroarylalkyl;

where each (C1- _C8 )alkyl, ( _C2 - _{C8)alkenyl, (C2-C8 ) alkynyl} , _C6 - _C20 -aryl, C3- _C20- carbocyclyl, _C2 - _C20 heterocyclyl, arylalkyl or heteroarylalkyl, optionally substituted with 1-3 R^^yn^M^ each R is, independently, H, (6-O^alkyl, (^-^alkenyl, (^-^alkynyl, C6-C20aryl, C ₃ -C ₂₀ -carbocyclyl, C ₂ -C ₂₀ -heterocyclyl or arylalkyl;

each R ¹¹ or R ¹² is, independently, H, (C1- _{C8 )} alkyl, ( _C2 - _C8 )alkenyl, ( _C2C8 )alkynyl, ( _C4 - _C8 )carbocyclylalkyl, optionally substituted aryl , optionally substituted heteroaryl, -C(=O)(C1- _C8 )alkyl, 4(O) _P (C1- _C8 )alkyl or aryl(C1- _C8 )alkyl;

each R ¹³ is, independently, a carbocycle or a heterocycle, optionally substituted with 1-3 R ²⁰ groups;

each R ²⁰ is, independently, halogen, CN, N3, N(R)2, OR, -SR, -S(O)R, -S(O)2R, -S(O)(OR), -S (O)2(OR), -C(=Y ¹ )R, -C(=Y ¹ )OR or C(=Y ¹ )N(R)2;

where each (^-^alkyl, (^-^alkenyl, (^-^alkynyl or aryl(C ₁ -C ₈ ))alkyl of each R ⁴ , R ¹¹ or R ¹² is, independently, optionally substituted one or more halos, hydroxy, CN, N3, N(R ^b )2 or OR ^b ; and wherein one or more non-terminal carbon atoms of each said (C1-C ₈ )alkyl may optionally be replaced by -O-, -S- or -NR ^b ;

each R ^b represents, independently, H, (^-^alkyl, (^-^alkenyl, (^-^alkynyl, aryl^-^alkyl, (C4-C8)carbocyclylalkyl, -C(=O)R ²¹ , -C(=O)OR ²¹ , -C(=O)NR ²¹ R ²² , -C(=O)SR ²¹ , -S(O)R ²¹ , -S(O)2R ²¹ , -S(O) (OR ²¹ ), -S(O)2(OR ²¹ ) or -SO2NR ²¹ R ²² ;

each R ²¹ or R ²² is, independently, H, (^-^alkyl, (^-^alkenyl, (C ₂ ^alkynyl, (C ₄ -C ₈ )carbocyclylalkyl, -C(=O)(C ₁ - C ₈ )alkyl, -S(O) _n (C ₁ -C ₈ )alkyl or aryl(C ₁ -C ₈ )alkyl and with the optional proviso that compounds 1, 1c, 1d, 1e, 2, TP-1; , A-1, 8 and 21 are excluded.

In another aspect of this embodiment, R ^a is H, (^-^alkyl or -C(=O)(C ₁ -C ₆ )alkyl; R ⁷ or R ⁷ together with R ⁴ is

where a is the point of attachment to R ⁷ ;

b is the point of attachment to R ⁴ ;

Ar represents phenyl or naphthyl, wherein phenyl and naphthyl are optionally substituted with 1-3 R ²⁰ groups;

each R ^y is, independently, (^-^alkyl or ^-^-carbocyclyl, where alkyl and carbocyclyl are optionally substituted with 1-3 R ²⁰ groups;

each R is independently H, (^-^alkyl or arylalkyl; and each ^R20 is independently halogen, CN, N(R)2, OR, -SR, -S(O)R, -S (O)2R, -S(O)(OR), -S(O) ₂ (OR), -C(=O)R, -C(=O)OR or C(=O)N(R) ₂ .

In another embodiment, the compounds of formulas IV-VI are compounds having the structure

- 16 046452

- 17 046452

or pharmaceutically acceptable salts thereof.

In one embodiment of Formulas I-III and Formulas IV-VI, R ¹¹ or R ¹² is independently H, (C1-C8)alkyl, ( _C2 -C8)alkenyl, ( _C2 -C8)alkynyl, (C ₄ -C ₈ )carbocyclylalkyl, optionally substituted aryl, optionally substituted heteroaryl, -C^OX^-C^alkyl, ^(O) _p (C1- _C8 )alkyl or aryl( _C1 - _C8 )alkyl. In another embodiment, R ¹¹ and R ¹² , taken together with the nitrogen to which they are both attached, form a 3- to 7-membered heterocycle, wherein any carbon atom of said heterocyclic ring may optionally be replaced by -O-, -S - or -NR ^a -. Thus, by way of example and not limitation, the -NR11R ¹² residue may be represented by heterocycles

and the like.

In another embodiment of Formulas I-III and Formulas IV-VI, each R ³ , R ⁴ , R ⁵ , R ⁶ , R ¹¹ or R ¹² is, independently, (C1-C8)alkyl, (C2-C8)alkenyl, (C2-C8)alkynyl or aryl (C1-C8)alkyl, wherein said (C1-C8)alkyl, (C2-C8)alkenyl, (C2-C8)alkynyl or aryl (C1-C8)alkyl, independently, is optionally substituted one or more halo, hydroxy, CN, N3, N(R ^a )2 or OR ^a . Thus, by way of example and not limitation, R ³ , R ⁴ , R ⁵ , R ⁶ , R ¹¹ or R ¹² may represent moieties such as -CH(NH2)CH3, -CH(OH)CH2CH3, -CH (NH2)CH(CH3)2, -CH2CF3, -(CH2)2CH(N3)CH3, -(CH2XNH2 and the like.

In another embodiment of Formulas I-III and Formulas IV-VI, R ³ , R ⁴ , R ⁵ , R ⁶ , R ¹¹ or R ¹² is (C1-C8)alkyl, wherein one or more non-terminal carbon atoms of each (C1C8 )alkyl optionally may be replaced by -O-, -S- or -NR ^a- . Thus, by way of example and not limitation, ^R3 , ^R4 , ^R5 , ^R6 , ^R11 , or ^R12 may represent moieties such as _-CH2OCH3 , -CH2OCH2CH3, -CH2OCH(CH3)2, -CH2SCH3 , -(CH2XOCH3, -(CH2)6N(CH3)2 and the like.

- 18 046452

In another embodiment, embodiments of formulas I-III are a compound selected from the group consisting of

- 19 046452

- 20 046452

- 21 046452

or a pharmaceutically acceptable salt or ester thereof.

- 22 046452

In another embodiment, a compound is provided useful for the synthesis of compounds of formula I selected from the group consisting of

or their salts or esters.

Definitions

Unless otherwise specified, the following terms and phrases used herein are intended to be understood with the following meanings.

When trademarks are used herein, it is the intention of applicants to independently include the trade name of the product and the active pharmaceutical ingredient(s) of the trademarked product.

As used herein, compounds of the invention or a compound of Formula I means a compound of Formula I or a pharmaceutically acceptable salt thereof. Likewise, with respect to isolated intermediates, the phrase compound of formula (number) means a compound of that formula and its pharmaceutically acceptable salts.

Alkyl is a hydrocarbon containing normal, secondary, tertiary or cyclic carbon atoms. For example, alkyl groups may have 1 to 20 carbon atoms (e.g., C1- _C20 alkyl), 1 to 8 carbon atoms (e.g., C1- _C8 alkyl), or 1 to 6 carbon atoms (e.g., C ₁ -C ₆ -alkyl). Examples of suitable alkyl groups include, but are not limited to, methyl (Me, -CH3), ethyl (Et, -CH2CH3), 1-propyl (n-PPr, n-propyl, -CH2CH2CH3), 2-propyl (i-Pr , i-propyl, CH(CH3) ₂ ), 1-butyl (n-Bu, n-butyl, -CH2CH2CH2CH3), 2-methyl-1-propyl (i-Bu, i-butyl, -CH2CH(CH3)2 ), 2-butyl (s-Bu, s-butyl, -CH(CH3)CH2CH3), 2-methyl-2-propyl (t-Bu, t-butyl, -C(CH3)3), 1-pentyl (n- pentyl, -CH2CH2CH2CH2CH3), 2-pentyl (-CH(CH ₃ )CH ₂ CH ₂ CH ₃ ), 3-pentyl (-CH(CH ₂ CH ₃ ) ₂ ), 2methyl-2-butyl (-C(CH ₃ ) ₂ CH ₂ CH ₃ ), 3-methyl-2-butyl (-CH(CH ₃ )CH(CH ₃ ) ₂ ), 3-methyl-1-butyl (-CH2CH2CH(CH3)2), 2-methyl- 1-butyl (-CH2CH(CH3)CH2CH3), 1-hexyl (-CH2CH2CH2CH2CH2CH3), 2hexyl (-CH(CH3)CH2CH2CH2CH3), 3 hexyl (-CH(CH2CH3)(CH2CH2CH3)), 2 methyl-2-pentyl ( -C(CH ₃ ) ₂ CH ₂ CH ₂ CH ₃ ), 3-methyl-2-pentyl (-CH(CH ₃ )CH(CH ₃ )CH ₂ CH ₃ ), 4-methyl-2-pentyl (-CH (CH3)CH2CH(CH3)2), 3-methyl-3-pentyl (-C(CH3)(CH2CH3)2), 2-methyl-3-pentyl (-CH(CH2CH3)CH(CH3)2), 2 ,3-dimethyl-2-butyl (-C(CH3)2CH(CH3)2), 3,3-dimethyl-2-butyl (-CH(CH3)C(CH3)3 and octyl (-(¾^¾) .

Alkoxy means a group having the formula -O-alkyl, in which the alkyl, as defined above, is attached to the parent molecule through an oxygen atom. The alkyl part of the alkoxy group may have from 1 to 20 carbon atoms (for example, C1-C ₂₀ -alkoxy from 1 to 12 carbon atoms (for example, C1C ₁₂ -alkoxy) or from 1 to 6 carbon atoms (for example, C1-C ₆ -alkoxy )- Examples of suitable alkoxy groups include, but are not limited to, methoxy (-O-CH3 or -OMe), ethoxy (-OCH2CH3 or OEt),

- 23 046452 tert-butoxy (-OC(CH ₃ )3 or -OtBu) and the like.

Haloalkyl is an alkyl group, as defined above, in which one or more hydrogen atoms of the alkyl group are replaced by halogen atoms. The alkyl portion of a haloalkyl group may have 1 to 20 carbon atoms (e.g., C.'i-C'20-haloalkyl), 1 to 12 carbon atoms (e.g., GC^-haloalkyl), or 1 to 6 carbon atoms (e.g. , Q-Sb-alkyl). Examples of suitable haloalkyl groups include, but are not limited to, -CF ₃ , -CHF ₂ , -CFH ₂ , -CH ₂ CF ₃ and the like.

Alkenyl is a hydrocarbon containing normal, secondary, tertiary or cyclic carbon atoms with at least one site of unsaturation, i.e. carbon-carbon, sp ² double bond. For example, an alkenyl group may have from 2 to 20 carbon atoms (for example, C ₂ -C ₂₀ -alkenyl), from 2 to 8 carbon atoms (for example, C ₂ -C ₈ -alkenyl) or 2 to 6 carbon atoms (for example, C ₂ -C ₆ -alkenyl). Examples of suitable alkenyl groups include, but are not limited to, ethylene or vinyl (-CH= _CH2 ), allyl ( _-CH2CH = _CH2 ), cyclopentenyl (-C5H7) and 5-hexenyl (-CH2CH2CH2CH2CH=CH2).

Alkynyl is a hydrocarbon containing normal, secondary, tertiary or cyclic carbon atoms with at least one site of unsaturation, i.e. carbon-carbon, sp triple bond. For example, an alkynyl may have from 2 to 20 carbon atoms (for example, C ₂ -C ₂₀ -alkynyl), from 2 to 8 carbon atoms (for example, C ₂ -C ₈ -alkynyl,) or from 2 to 6 carbon atoms (for example , C ₂ -C ₆ alkynyl). Examples of suitable alkynyl groups include, but are not limited to, acetylene (-C^CH), propargyl (-CH ₂ C=CH) and the like.

Alkylene refers to a saturated, branched or straight chain or cyclic hydrocarbon radical having two monovalent radical centers obtained by removing two hydrogen atoms from the same or two carbon atoms of the parent alkane. For example, an alkylene group may have 1 to 20 carbon atoms, 1 to 10 carbon atoms, or 1 to 6 carbon atoms. Typical alkylene radicals include, but are not limited to, methylene (-CH2-), 1,1-ethyl (-CH(CH3)-), 1,2-ethyl (-CH2CH2-), 1,1-propyl (-CH (CH2CH3)-), 1,2-propyl (-CH ₂ CH(CH ₃ )-), 1,3-propyl (-CH2CH2CH2-), 1,4-butyl (-CH ₂ CH ₂ CH ₂ CH ₂ - ) etc.

Alkenylene refers to an unsaturated, branched or straight chain or cyclic hydrocarbon radical having two monovalent radical centers obtained by removing two hydrogen atoms from the same or two carbon atoms of the parent alkene. For example, an alkenylene group may have 1 to 20 carbon atoms, 1 to 10 carbon atoms, or 1 to 6 carbon atoms. Typical alkenylene radicals include, but are not limited to, 1,2ethylene(-CH=CH-).

Alkynylene refers to an unsaturated, branched or straight chain or cyclic hydrocarbon radical having two monovalent radical centers obtained by removing two hydrogen atoms from the same or two carbon atoms of the parent alkyne. For example, an alkynylene group may have 1 to 20 carbon atoms, 1 to 10 carbon atoms, or 1 to 6 carbon atoms. Typical alkynylene radicals include, but are not limited to, acetylene (-C=C-), propargyl (-CH ₂ C^C-) and 4-pentynyl (-CH ₂ CH ₂ CH ₂ C=C-).

Amino refers generally to a nitrogen radical which may be considered an ammonia derivative having the formula -N(X) ₂ wherein each X is, independently, H, substituted or unsubstituted alkyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, etc. .d. The nitrogen hybridization is about sp ³ . Non-limiting types of amino include -NH2, -N(alkyl) ₂ , -NH(alkyl), -N(carbocyclyl) ₂ , -NH(carbocyclyl), -N(heterocyclyl) ₂ , -NH(heterocyclyl), -N(aryl) ) ₂ , -NH(aryl), -Н(alkyl)(aryl), -Н(alkyl)(heterocyclyl),

-N(carbocyclyl)(heterocyclyl), -N(aryl)(heteroaryl), -N(alkyl)(heteroaryl), etc. The term alkylamino refers to an amino group substituted with at least one alkyl group. Non-limiting examples of amino groups include -NH ₂ , -NH(CH ₃ ), -N(CH ₃ ) ₂ , -NH(CH ₂ CH ₃ ), -N(CH ₂ CH ₃ ) ₂ , -NH(phenyl), -No. (phenyl) ₂ , -KH(benzyl), -N(benzyl) ₂ , etc. Substituted alkylamino generally refers to alkylamino groups, as defined above, in which at least one substituted alkyl, as defined herein, is attached to an amino nitrogen atom. Non-limiting examples of substituted alkylamino include -MH(alkylene-C(O)-OH), -UN(alkylene-C(O)-O-alkyl), -N(alkylene-C(O)-OH) ₂ , - N(alkylene-C(O)-O-alkyl) ₂ , etc.

Aryl means an aromatic hydrocarbon radical obtained by removing one hydrogen atom from one carbon atom of the aromatic parent ring system. For example, an aryl group may have 6 to 20 carbon atoms, 6 to 14 carbon atoms, or 6 to 10 carbon atoms. Typical aryl groups include, but are not limited to, radicals derived from benzene (eg, phenyl), substituted benzene, naphthalene, anthracene, diphenyl, and the like.

Aryl alkyl refers to an acyclic alkyl radical in which one of the hydrogen atoms bonded to a carbon atom, typically a terminal or sp ³ carbon atom, is replaced by an aryl radical. Typical arylalkyl groups include, but are not limited to, benzyl, 2-phenylethan-1-yl,

- 24 046452 naphthylmethyl, 2-naphthylethan-1-yl, naphthobenzyl, 2-naphthophenylethan-1-yl and the like. An arylalkyl group may include from 7 to 20 carbon atoms, for example, the alkyl portion is from 1 to 6 carbon atoms, and the aryl portion is from 6 to 14 carbon atoms.

Arylalkenyl refers to an acyclic alkenyl radical in which one of the hydrogen atoms bonded to a carbon atom, usually a terminal or sp ³ carbon atom, but also an sp ² carbon atom, is replaced by an aryl radical. The aryl moiety of arylalkenyl may include, for example, any of the aryl groups described herein, and the alkenyl moiety of arylalkenyl may include, for example, any of the alkenyl groups described herein. An arylalkenyl group may contain from 8 to 20 carbon atoms, for example, an alkenyl moiety of 2 to 6 carbon atoms and an aryl moiety of 6 to 14 carbon atoms.

Arylalkynyl refers to an acyclic alkynyl radical in which one of the hydrogen atoms bonded to a carbon atom, typically a terminal or sp ³ carbon atom, as well as an sp carbon atom, is replaced by an aryl radical. The aryl moiety of arylalkynyl may include, for example, any of the aryl groups described herein, and the alkynyl moiety of arylalkynyl may include, for example, any of the alkynyl groups described herein. An arylalkynyl group may contain from 8 to 20 carbon atoms, for example an alkynyl moiety of 2 to 6 carbon atoms and an aryl moiety of 6 to 14 carbon atoms.

The term substituted refers to alkyl, alkylene, aryl, arylalkyl, alkoxy, heterocyclyl, heteroaryl, carbocyclyl, etc., for example, substituted alkyl, substituted alkylene, substituted aryl, substituted arylalkyl, substituted heterocyclyl and substituted carbocyclyl, unless otherwise specified, means alkyl, alkylene, aryl, arylalkyl, heterocyclyl, carbocyclyl, respectively, in which one or more hydrogen atoms, independently of each other, are replaced by a non-hydrogen substituent. Typical substituents include, but are not limited to, -X, -R ^b , -O ^- , =O, -OR ^b , -SR ^b , -S ^- , -NR ^b 2, -N+R ^b 3, =NR ^b , -CX3, -CN, -OCN, -SCN, -N=C=O, -NCS, -NO, -NO2, =N, -N3, -NHC(=O)R ^b , -OC(=O) R ^b , -NHC(=O)NR ^b 2, -S(=O)2-, -S(=O)2OH, -S(=O)2R ^b , -OS(=O)2OR ^b ,

-S(=O)2NR ^b 2, -S(=O)R ^b , -OP(=O)(OR ^b )2, -P(=O)(OR ^b )2, -P(=O)( O ^- )2, -P(=O)(OH)2, -P(O)(OR ^b )(O ^- ), -C(=O)R ^b , -C(=O)X, -C( S)R ^b , -C(O)OR ^b , -C(O)O ^- , -C(S)OR ^b , -C(O)SR ^b , -C(S)SR ^b , -C(O) NR ^b 2, -C(S)NR ^b 2, -C(=NR ^b )NR ^b ₂ where each X is independently a halogen: F, Cl, Br or I; and each R ^b is independently H, alkyl, aryl, arylalkyl, heterocycle or a protecting group, or a prodrug moiety. Alkylene, alkenylene and alkynylene groups can also be easily substituted. Unless otherwise indicated, when the term substituted is used in conjunction with groups such as arylalkyl that have two or more substituent moieties, the substituents may be attached to the aryl moiety, the alkyl moiety, or both.

The term prodrug as used herein refers to any compound that, when introduced into a biological system, generates a drug substance, i.e. active ingredient, as a result of spontaneous chemical reaction(s), enzymatically catalyzed chemical reaction(s), photolysis and/or metabolic chemical reaction(s). A prodrug is thus a covalently modified analogue or latent form of a therapeutically active compound.

Those skilled in the art will appreciate that substituents and other moieties of the compounds of Formulas I-III and Formulas IV-VI must be selected to provide a compound that is sufficiently stable to provide pharmaceutically useful compounds that can be combined into acceptably stable pharmaceutical composition. The definitions and substituents of the various genus and subgenus of the present compounds are described and illustrated herein. It will be understood by one skilled in the art that any combinations of terms and substituents described above should not render the species or compound inoperable. Inoperable species or compound means a combination of structures that violate relevant scientific principles (such as, for example, a carbon atom attached to more than four covalent bonds) or compounds that are too unstable to allow isolation and combination into pharmaceutically acceptable dosage forms.

Heteroalkyl refers to an alkyl group in which one or more carbon atoms are replaced by a heteroatom, such as O, N, or S. For example, if the carbon atom of an alkyl group that is attached to the parent molecule is replaced by a heteroatom (such as O, N, or S), the resulting heteroalkyl group is, respectively, an alkoxy group (eg OCH ₃ etc.), an amine (eg -NHCH ₃ , -N(CH ₃ ) ₂ etc.) or a thioalkyl group (eg -SCH ₃ ). If the non-terminal carbon atom of an alkyl group that is not attached to the parent molecule is replaced by a heteroatom (for example, O, N or S), the resulting heteroalkyl group is, respectively, an alkyl ether (for example, -CH ₂ CH ₂ -O-CH ₃ etc. .d.), alkylamine (for example, -CH2NHCH3, -CH ₂ N(CH ₃ ) ₂ , etc.) or thioalkyl ether (for example, -CH2-S-CH3). If the terminal carbon atom of an alkyl group is replaced by a heteroatom (for example, O, N or S), the resulting heteroalkyl group is, respectively, a hydroxyalkyl group (for example, -CH2CH2-OH), an aminoalkyl group (for example, -CH ₂ NH2) or an alkyl thiol group. group (for example, -CH ₂ CH ₂ -SH). A heteroalkyl group may have, for example, 1 to 20 carbon atoms, 1 to 10 carbon atoms, or 1 to 6 carbon atoms.

- 25 046452

Ci-Cb-heteroalkyl group means a heteroalkyl group having from 1 to 6 carbon atoms.

Heterocycle or heterocyclyl as used herein includes by way of example, but not limitation, heterocycles described in Paquette, Leo A.; Principles of Modern Heterocyclic Chemistry (W.A. Benjamin, New York, 1968), especially chapters 1, 3, 4, 6, 7 and 9; The Chemistry of Heterocyclic Compounds, A Series of Monographs (John Wiley & Sons, New York, 1950 to present), particularly Volumes 13, 14, 16, 19 and 28, and J. Am. Chem. Soc. (1960) 82:5566. In one specific embodiment, a heterocycle includes a carbocycle, as defined herein, in which one or more (eg, 1, 2, 3, or 4) carbon atoms have been replaced by a heteroatom (eg, O, N, or S). The term heterocycle or heterocyclyl includes saturated rings, partially unsaturated rings and aromatic rings (ie, heteroaromatic rings). Substituted heterocycles include, for example, heterocyclic rings substituted with any of the substituents described herein, including carbonyl groups. A non-limiting example of a carbonyl substituted heterocyclyl is

Examples of heterocycles include, by way of example and not limitation, pyridyl, dihydropyridyl, tetrahydropyridyl (piperidyl), thiazolyl, tetrahydrothiophenyl, sulfur-oxidized tetrahydrothiophenyl, pyrimidinyl, furanyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, tetrazolyl, benzofuranyl, thianaphthalenyl, indolyl, lazy , quinolinyl, isoquinolinyl, benzimidazolyl, piperidinyl, 4-piperidonyl, pyrrolidinyl, 2-pyrrolidonyl, pyrrolinyl, tetrahydrofuranyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, octahydroisoquinolinyl, azocinyl, triazinyl, diazinyl, 2H,6H-1,5,2 -dithiazinyl, thienyl, thianthrenyl, pyranyl, isobenzofuranyl, chromenyl, xanthenyl, phenoxatinyl, 2H-pyrrolyl, isothiazolyl, isoxazolyl, pyrazinyl, pyridazinyl, indolizinyl, isoindolyl, 3H-indolyl, 1H-indazolyl, purinyl, 4H-quinolizinyl, phthalazinyl, iridinyl , quinoxalinyl, quinazolinyl, cinnolinyl, pteridinyl, 4aH-carbazolyl, carbazolyl, β-carbolinyl, phenanthridinyl, acridinyl, pyrimidinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, furazanil, phenoxazinyl, isochromanil, chromanyl, imidazolidinyl, imidazolinyl, dinyl, pyrazolinil, piperazinil, indolinyl , isoindolinyl, quinuclidinyl, morpholinyl, oxazolidinyl, benzotriazolyl, benzisoxazolyl, oquindolyl, benzoxazolinyl, isatinoyl and bis-tetrahydrofuranyl

By way of example and not limitation, carbon-linked heterocycles are bonded at the 2, 3, 4, 5, or 6 position of pyridine, the 3, 4, 5, or 6 position of pyridazine, the 2, 4, 5, or 6 position of pyrimidine, the 2, 3 position , 5 or 6 pyrazine, 2, 3, 4 or 5 position of furan, tetrahydrofuran, thiofuran, thiophene, pyrrole or tetrahydropyrrole, 2, 4 or 5 position of oxazole, imidazole or thiazole, 3, 4 or 5 position of isoxazole, pyrazole or isothiazole, the 2 or 3 position of aziridine, the 2, 3 or 4 position of azetidine, the 2, 3, 4, 5, 6, 7 or 8 position of quinoline or the 1, 3, 4, 5, 6, 7 or 8 position of isoquinoline. Even more typically carbon-linked heterocycles include 2-pyridyl, 3-pyridyl, 4-pyridyl, 5-pyridyl, 6pyridyl, 3-pyridazinyl, 4-pyridazinyl, 5-pyridazinyl, 6-pyridazinyl, 2-pyrimidinyl, 4-pyrimidinyl, 5- pyrimidinyl, 6-pyrimidinyl, 2-pyrazinyl, 3-pyrazinyl, 5-pyrazinyl, 6-pyrazinyl, 2-thiazolyl, 4-thiazolyl or 5-thiazolyl.

By way of example and not limitation, nitrogen-linked heterocycles are bonded at the 1-position of aziridine, azetidine, pyrrole, pyrrolidine, 2-pyrroline, 3-pyrroline, imidazole, imidazolidine, 2-imidazoline, 3-imidazoline, pyrazole, pyrazoline, 2-pyrazoline, 3-pyrazoline, piperidine, piperazine, indole, indoline, 1H-indazole, 2-position of isoindole or isoindoline, 4-position of morpholine and 9-position of carbazole or β-carboline. Even more typically nitrogen-linked heterocycles include 1-aziridyl, 1-azetedyl, 1-pyrrolyl, 1-imidazolyl, 1-pyrazolyl, 1-piperidinyl.

Heterocyclylalkyl refers to an acyclic alkyl in which one of the hydrogen atoms bonded to a carbon atom, typically a terminal or sp ³ carbon atom, is replaced by a heterocyclyl radical (ie, a heterocyclylalkylene moiety). Typical heterocyclyl-alkyl groups include, but are not limited to, heterocyclyl-OH^, 2-(heterocyclyl)ethan-1-yl, and the like, wherein the heterocyclyl moiety includes any of the heterocyclyl groups described above, including including those described in the Principles of Modern Heterocyclic Chemistry. Those skilled in the art will also appreciate that a heterocyclyl group can be attached to the alkyl moiety of a heterocyclyl alkyl via carbon-carbon or carbon-heteroatom bonds, so long as the resulting group is chemically stable. A heterocyclyl alkyl group contains from 3 to 20 carbon atoms, for example, the alkyl part of an arylalkyl group has from 1 to 6 carbon atoms, and the heterocyclyl part has from 2 to 14 carbon atoms. Examples of heterocyclylalkyls include

- 26 046452 by way of example and not limitation, 5-membered sulfur, oxygen and/or nitrogen-containing heterocycles such as thiazolylmethyl, 2-thiazolyl-1-yl, imidazolylmethyl, oxazolylmethyl, thiadiazolylmethyl, etc., 6-membered sulfur, oxygen and/or nitrogen-containing heterocycles such as piperidinylmethyl, piperazinylmethyl, morpholinylmethyl, pyridinylmethyl, pyridizylmethyl, pyrimidylmethyl, pyrazinylmethyl, etc.

Heterocyclylalkenyl refers to an acyclic alkenyl radical in which one of the hydrogen atoms bonded to a carbon atom, typically a terminal or sp ³ carbon atom, as well as an sp ² carbon atom is replaced by a heterocyclyl radical (ie, a heterocyclyl alkenylene moiety). The heterocyclyl portion of a heterocyclylalkenyl group includes any of the heterocyclyl groups described herein, including those described in the Principles of Modern Heterocyclic Chemistry, and the alkenyl portion of the heterocyclylalkenyl group includes any of the alkenyl groups described herein. Those skilled in the art will also appreciate that a heterocyclyl group can be attached to the alkenyl moiety of a heterocyclyl-alkenyl via carbon-carbon or carbon-heteroatom bonds, so long as the resulting group is chemically stable. A heterocyclyl-alkenyl group has 4 to 20 carbon atoms, for example, the alkenyl portion of a heterocyclylalkenyl group has 2 to 6 carbon atoms, and the heterocyclyl portion has 2 to 14 carbon atoms.

Heterocyclylalkynyl refers to an acyclic alkynyl radical in which one of the hydrogen atoms bonded to a carbon atom, typically a terminal or sp ³ carbon atom, as well as the sp carbon atom is replaced by a heterocyclyl radical (ie, a heterocyclyl alkynylene moiety). The heterocyclyl portion of a heterocyclyl-alkynyl group includes any of the heterocyclyl groups described herein, including those described in the Principles of Modern Heterocyclic Chemistry, and the alkynyl portion of the heterocyclyl-alkynyl group includes any of the alkynyl groups described herein. Those skilled in the art will also appreciate that a heterocyclyl group can be attached to the alkynyl moiety of a heterocyclylalkynyl via carbon-carbon or carbon-heteroatom bonds, provided that the resulting group is chemically stable. A heterocyclylalkynyl group has 4 to 20 carbon atoms, for example, the alkynyl portion of a heterocyclylalkynyl group has 2 to 6 carbon atoms, and the heterocyclyl portion has 2 to 14 carbon atoms.

Heteroaryl refers to an aromatic heterocyclyl having at least one heteroatom on the ring. Non-limiting examples of suitable heteroatoms that may be included on the aromatic ring include oxygen, sulfur and nitrogen. Non-limiting examples of a heteroaryl ring include all aromatic rings listed in the definition of heterocyclyl, including pyridinyl, pyrrolyl, oxazolyl, indolyl, isoindolyl, purinyl, furanyl, thienyl, benzofuranyl, benzothiophenyl, carbazolyl, imidazolyl, thiazolyl, isoxazolyl, pyrazolyl, isothiazolyl , quinolyl, isoquinolyl, pyridazyl, pyrimidyl, pyrazyl, etc.

Carbocycle or carbocyclyl refers to a saturated (e.g., cycloalkyl), partially unsaturated (e.g., cycloalkenyl, cycloalkadienyl, etc.) or aromatic ring having from 3 to 7 carbon atoms as a monocycle, from 7 to 12 carbon atoms as a bicycle, and up to up to 20 carbon atoms as a polycycle. Monocyclic carbocycles have 3 to 7 ring atoms, even more typically 5 or 6 ring atoms. Bicyclic carbocycles have from 7 to 12 atoms in the ring, for example, they are arranged as bicyclo [4,5], [5,5], [5,6] or [6,6] systems, or 9 or 10 atoms in the ring are arranged as bicyclo [5,6] or [6,6] systems, or spiro-fused rings. Non-limiting examples of monocyclic carbocycles include cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclohexyl, 1-cyclohex-1-enyl, 1-cyclohex-2 -enyl, 1cyclohex-3-enyl and phenyl. Non-limiting examples of bicyclo carbocycles include naphthyl, tetrahydronaphthalene and decalin.

Carbocyclylalkyl refers to an acyclic alkyl in which one of the hydrogen atoms bonded to the carbon atom is replaced by carbocyclyl, as described herein. Typical, but non-limiting examples of a carbocyclylalkyl group include cyclopropylmethyl, cyclopropylethyl, cyclobutylmethyl, cyclopentylmethyl and cyclohexylmethyl.

Arylheteroalkyl refers to a heteroalkyl, as defined herein, in which a hydrogen atom (which can be attached to either a carbon atom or a heteroatom) has been replaced by an aryl group, as defined herein. Aryl groups may be bonded to a carbon atom of a heteroalkyl group or a heteroatom of a heteroalkyl group, provided that the resulting arylheteroalkyl group provides a chemically stable moiety. For example, an arylheteroalkyl group may have the general formulas alkyleneI-aryl, -alkylene-O-αalkylene-aryl, alkylene-NH-aryl, -alkylene-NH-alkylenaryl, -alkylene-aryl, -alkylene-NH-alkylene-aryl, etc. d. In addition, any of the alkylene moieties in the general formula above may be further substituted by any of the substituents defined or given herein.

Heteroarylalkyl refers to an alkyl group, as defined herein, in which the hydrogen atom is replaced by a heteroaryl group, as defined herein. Non-limiting Examples of Heteroarylalk

- 27 046452 la include -CH ₂ -pyridinyl, -CH ₂ -pyrrolyl, -CH ₂ -oxazolyl, -CH ₂ -indolyl, -CH ₂ -isoindolyl, -CH2purinyl, -CH ₂ -furanyl, -CH ₂ -thienyl, -CH ₂ -benzofuranyl, -CH ₂ -benzothiophenyl, -CH ₂ -carbazolyl, -CH2-imidazolyl, -CH _{2 -thiazolyl, -CH 2 -isoxazolyl} , -CH ₂ -pyrazolyl, -CH ₂ -isothiazolyl, -CH ₂ -quinolyl , -CH2isoquinolyl, _-CH2 -pyridazyl, _-CH2 -pyrimidyl, -CH2 _- pyrazyl, -CH( _CH3 )-pyridinyl, -CH( _CH3 )pyrrolyl, -CH( _CH3 )-oxazolyl, -CH (CH ₃ )-indolyl, -CH(CH ₃ )-isoindolyl, -CH(CH ₃ )-purinyl, -CH(CH ₃ )-furanyl, -CH(CH ₃ )-thienyl, -CH(CH ₃ )- benzofuranyl, -CH(CH ₃ )-benzothiophenyl, -CH(CH ₃ )carbazolyl, -CH(CH ₃ )-imidazolyl, -CH(CH ₃ )-thiazolyl, -CH(CH ₃ )-isoxazolyl, -CH(CH ₃ )-pyrazolyl, -CH(CH ₃ )-isothiazolyl, -CH(CH ₃ )-quinolyl, -CH(CH ₃ )-isoquinolyl, -CH(CH ₃ )-pyridazyl, -CH(CH ₃ )-pyrimidyl, -CH(CH ₃ )-pyrazyl, etc.

The term optionally substituted, with reference to a particular moiety of a compound of Formulas I-III and Formulas IV-VI (e.g., an optionally substituted aryl group), refers to a moiety in which all of the substituents are hydrogen or in which one or more of the hydrogens may be partially replaced by substituents such as those listed under the definition of substituted, or as otherwise indicated.

The term is optionally substituted with reference to a specific moiety of a compound of Formulas I-III and Formulas IV-VI (e.g., the carbon atoms of said (C ₁ -C ₈ )αalkyl may optionally be replaced by -O-, -S- or -NR ^a- ) , means that one or more (C ₁ -C ₈ )αalkyl methylene groups can be replaced by 0, 1, 2 or more of these groups (eg, -O-, -S- or -NR ^a -).

The term non-terminal carbon atom(s), with reference to an alkyl, alkenyl, alkynyl, alkylene, alkenylene or alkynylene moiety, refers to the carbon atoms in the moiety that are placed between the first carbon atom of the part and the last carbon atom of the part. Therefore, by way of example and not limitation, in the alkyl moiety -CH ₂ (C*)H ₂ (C*)H ₂ CH ₃ or the alkylene group -CH ₂ (C*)H ₂ (C*)H ₂ CH ₂ -C* atoms will be considered non-terminal carbon atoms.

Some Y and Y ¹ alternatives are nitrogen oxides, such as +N(O)(R) or +N(O)(OR). These nitrogen oxides, as shown here attached to a carbon atom, may also be represented by charge separated groups such as

Oh oh

respectively, and are intended to equivalently represent the above groups for purposes of describing the present invention.

Linker or bond means a fragment of a chemical substance containing a covalent bond or chain of atoms. Linkers include alkyloxy (eg, polyethyleneoxy, PEG, polymethyleneoxy) and alkylamino (eg, polyethyleneamino, Jeffamine™) units; as well as dibasic acid esters and amides, including succinate, succinamide, diglycolate, malonate and caproamide.

Terms such as oxygen-linked, nitrogen-linked, carbon-linked, sulfur-linked, or phosphorus-linked mean that if a bond between two moieties can be formed by more than one type of atom in a group, then the bond is formed between the moieties through the specified atom. For example, a nitrogen-linked amino acid may be linked through the nitrogen atom of the amino acid rather than through the oxygen or carbon atom of the amino acid.

Unless otherwise indicated, the carbon atoms of compounds of formulas I-III and formulas IV-VI are intended to have a valence of four. In some chemical representations of structures where the carbon atoms do not have enough variable bonding sites to produce a valence of four, the remaining carbon substituents needed to provide a valence of four should be considered hydrogen. For example,

R ⁴ f has the same meaning as f

A protecting group refers to a moiety of a compound that masks or changes the properties of functional groups or the properties of the compound as a whole. The chemical basis of the protecting group varies widely. One of the functions of a protecting group is to serve as an intermediate in the synthesis of the parent drug. Chemical protecting groups and protection/deprotection strategies are well known in the art. See: Protective Groups in Organic Chemistry, Theodora W. Greene (John Wiley & Sons, Inc., New York, 1991). Protective groups hour

- 28 046452 are then used to mask the reactivity of certain functional groups, to assist in the efficiency of desired chemical reactions, such as the formation and breaking of chemical bonds in the resulting and planned design. Protection of functional groups of compounds changes physical properties other than the reactivity of the protected functional groups, such as polarity, lipophilicity (hydrophobicity), and other properties that can be measured by common analytical instruments. Chemically protected intermediates may themselves be biologically active or inactive.

Protected compounds may also exhibit alternative, and in some cases optimized, properties in vitro and in vivo, such as passage through cell membranes and resistance to enzymatic degradation or uptake. In this role, protected compounds with putative therapeutic effects can be considered prodrugs. Another function of the protecting group is to convert the parent drug into a prodrug, causing the parent drug to release prodrug conversion in vivo. Because active prodrugs can be absorbed more efficiently than the parent drug, the prodrug may have greater in vivo efficacy than the parent drug. Protecting groups are removed either in vitro in the case of chemical intermediates or in vivo in the case of prodrugs. In the case of chemical intermediates, it is not particularly important that the resulting products after deprotection of, for example, alcohols, are physiologically acceptable, although in general it is more desirable that the products are pharmacologically harmless.

A prodrug moiety means a labile functional group that is separated from the active inhibitory compound by metabolism, systemically, intracellularly, by hydrolysis, enzymatic degradation, or some other process (Bundgaard, Hans, Design and Application of Prodrugs in Textbook of Drug Design and Development (1991), P. Krogsgaard-Larsen and H. Bundgaard, Eds. Harwood Academic Publishers, pp. 113-191). Enzymes that are capable of enzymatically activating the phosphonate compound prodrug mechanism of the invention include, but are not limited to, amidases, esterases, microbial enzymes, phospholipases, cholinesterases and phosphases. Prodrug moieties can help increase solubility, absorption, and lipophilicity to optimize drug delivery, bioavailability, and efficacy.

The prodrug moiety may include an active metabolite or the drug itself.

Examples of prodrug moieties include hydrolytically sensitive or labile alkoxymethyl ethers -CH2OC(=O)R ³⁰ and acyloxymethyl carbonates -CH2OC(=O)OR ³⁰ , where R ³⁰ is Cj-C.'6-alkyl. C1- _C6 substituted alkyl, ^-^-aryl or _C6 - _C20 substituted aryl. The acyloxyalkyl ester was used as a prodrug strategy for carboxylic acids and then applied to phosphates and phosphonates by Farquhar et al. (1983) J. Pharm. Sci. 72: 324; also US patent numbers 4816570, 4968788, 5663159 and 5792756. In some compounds of the invention, the prodrug moiety is part of a phosphate group. The acyloxyalkyl ester can be used to deliver phosphoric acids across cell membranes and increase oral bioavailability. A closed variant of the acyloxyalkyl ether, the alkoxycarbonyloxyalkyl ester (carbonate), may also enhance bioavailability as a prodrug moiety in the combination compounds of the invention. An exemplary acyloxymethyl ether is pivaloyloxymethoxy, (POM)-CH ₂ OC(=O)C(CH ₃ ) ₃ . An exemplary acyloxymethyl carbonate moiety of a prodrug is pivaloyloxymethyl carbonate (POC)-CH ₂ OC(=O)OC(CH ₃ ) ₃ .

The phosphate group may be part of a phosphate prodrug. The prodrug moiety may be sensitive to hydrolysis, such as, but not limited to, those containing pivaloyloxymethyl carbonate (POC) or POM groups. In addition, the prodrug moiety may be susceptible to enzymatically potentiated cleavage, such as a lactate ester or phosphonamidate ester group.

Phosphorus aryl esters, particularly phenyl esters, are reported to have increased oral bioavailability (DeLambert et al. (1994) J. Med Chem. 37:498). Phenyl ethers containing an ortho to phosphate carboxyl ester have also been described (Khamnei and Torrence, (1996) J. Med. Chem. 39:4109-4115). Benzyl esters are reported to generate the parent phosphonic acid. In some cases, substituents in the ortho or para position can lead to accelerated hydrolysis. Benzyl analogues with acylated phenols or alkylated phenols can generate phenolic compounds under the action of enzymes, for example, esterases, oxidases, etc., which, in turn, undergo cleavage at the C-O benzyl bond to produce phosphoric acid and quinone methide intermediates. Examples of this class of prodrugs are described by Mitchell et al (1992) J. Chem. Soc. Perkin Trans. I 2345; Brook et al WO 91/19721. Still other benzylic prodrugs have been described as containing a carboxyl ester moiety linked to a benzyl methylene (Glazier et al WO 91/19721). Thio prodrugs have been reported to be useful for the intracellular delivery of phosphonate drugs. These pro-esters contain an ethylthio group, in which the thiol group is either esterified with an acyl group or combined with another thiol group to form a disulfide. Deesterification or disulfide reduction generates a free thio intermediate

- 29 046452 duct, which subsequently decomposes into phosphoric acid and episulfide (Puech et al (1993) Antiviral Res., 22: 155-174; Benzaria et al (1996) J. Med. Chem. 39:4958). Cyclic phosphonate esters have also been described as prodrugs of phosphorus-containing compounds (Erion et al., US Pat. No. 6,312,662).

It should be noted that all enantiomers, diastereomers and racemic mixtures, tautomers, polymorphs, pseudopolymorphs of the compounds within Formula I, Formula II, Formula III, Formula IV, Formula V or Formula VI and their pharmaceutically acceptable salts are covered by the present invention. All mixtures of such enantiomers and diastereomers are within the scope of the present invention.

The compound of formulas I-III and formulas IV-VI and its pharmaceutically acceptable salts may exist in the form of various polymorphs or pseudo-polymorphs. As used here, crystal polymorphism refers to the ability of a crystalline compound to exist in different crystal structures. Crystal polymorphism can result from differences in crystal packing (packaging polymorphism) or differences in the packing of different conformers of the same molecule (conformational polymorphism). As used here, crystalline pseudopolymorphism refers to the ability of a hydrate or solvate of a compound to exist in different crystal structures. Pseudopolymorphs of the present invention may exist due to differences in crystal packing (packaging polymorphism) or due to differences in packing of different conformers of the same molecule (conformational polymorphism). The present invention includes all polymorphic and pseudopolymorphic compounds of formulas I-III and formulas IV-VI and their pharmaceutically acceptable salts.

The compound of formulas I-III and formulas IV-VI and its pharmaceutically acceptable salts may also exist in the form of an amorphous solid. As used here, an amorphous solid is a solid in which there is no long-range order in the arrangement of atoms in the solid. This definition applies when the crystal size is two nanometers or smaller. Additives, including solvents, can be used to create the amorphous forms of the present invention. The present invention includes all amorphous forms of the compounds of formulas I-III and formulas IV-VI and their pharmaceutically acceptable salts.

Selected substituents containing compounds of formulas I-III and formulas IV-VI are present in a recursive degree. In this context, a recursive proxy means that a proxy can have another instance similar to itself. Due to the recursive nature of such substituents, in principle, a large number of compounds may be present in any embodiment. For example, R ^x includes the substituent R ^y . R ^y could be R. R could be W ³ . W ³ may be W ⁴ and W ⁴ may be R or contain substituents containing R ^y One skilled in the art of medicinal chemistry will understand that the total number of such substituents is reasonably limited by the desired properties of the intended compound. Such properties include, by way of example and not limitation, physical properties such as molecular weight, solubility or log P, application properties such as activity on the intended target, and practical properties such as ease of synthesis.

By way of example and not limitation, W ³ and R ^y are recursive substituents in some embodiments. As a rule, each recursive replacement can occur independently of each other 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1 or 0 times in this embodiment. More typically, each recursive replacement may occur independently 12 or fewer times in a given embodiment. Even more typically, each recursive replacement may occur independently 3 or fewer times in a given embodiment. For example, W ³ will occur 0 to 8 times, R ^y will occur 0 to 6 times in this embodiment. Even more typically, W ³ will occur 0 to 6 times and R ^y will occur 0 to 4 times in this embodiment.

Recursive substituents are included within an aspect of the invention. One skilled in the art of medicinal chemistry will appreciate the versatility of such substituents. To the extent that recursive substituents are present in an embodiment of the invention, the total amount will be determined as above.

The modifier about is used in connection with quantity and includes specified values, and has a meaning dictated by the context (for example, it includes the degree of error associated with measuring a specific quantity).

Compounds of formulas I-III and formulas IV-VI may contain phosphate groups, such as R ⁷ , which may be a prodrug moiety

where each Y or Y ¹ is, independently, O, S, NR, ⁺ N(O)(R), N(OR), ⁺ N(O)(OR) or N-NR ₂ ; W ¹ and

- 30 046452

W ² taken together are -Y ³ (C(R ^y ) ₂ ) ₃ Y ³ -; or one of W ¹ or W ² , together with any R ³ or R ⁴ , is -Y ³ - and the other W ¹ or W ² is Formula Ia, or W ¹ and W ² are each independently a group of Formula Ia

where each Y ² represents, independently, a bond, O, CR2, NR, +N(O)(R), N(OR), ⁺ N(O)(OR), NNR2, S, SS, S(O) or S(O)2;

each Y ³ is independently O, S or NR;

M2 represents 0, 1 or 2;

each R ^y represents, independently, H, F, Cl, Br, I, OH, R, -C(=Y1)R, -C(=Y1)OR, -C(=Y1)N(R)2, -N(R)2, - ⁺ N(R)3, -SR, -S(O)R, -S(O)2R -S(O)(OR), -S(O)2(OR), -OC(=Y1)R, -OC(=Y1)OR, -OC(=Y1)(N(R)2), -SC(=Y ¹ )R, -SC(=Y ¹ )OR, -SC (=Y1)(N(R)2), -N(R)C(=Y1)R, -N(R)C(=Y1)OR or -N(R)C(=Y1)N(R) 2, -SO ₂ NR ₂ , -CN, -N ₃ , -NO ₂ , -OR, protecting group or W ³ ; or, taken together, two ^Ry on the same carbon atoms form carbocyclic rings of from 3 to 7 carbon atoms;

each R ^x represents, independently, R ^y , a protecting group or the formula

where M1a, M1c and M1d are independently 0 or 1;

M12c represents 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12;

each R is H, halogen, (C1- _C8 )alkyl, (C1- _C8 )substituted alkyl, ( _C2 - _C8 )alkenyl, ( _C2 - _C8 )substituted alkenyl, ( _C2 - _C8 )alkynyl, (C ₂ -C ₈ ) substituted alkynyl, C ₆ -C ₂₀ -aryl, C ₆ -C ₂₀ substituted aryl, C ₂ -C ₂ o-heterocycle, C ₂ -C ₂₀ substituted heterocyclyl, arylalkyl, substituted arylalkyl or a protecting group;

W ³ is W ⁴ or W ⁵ ; W ⁴ represents R, -C(Y1)R ^y , -C(Y1)W ⁵ , -SO2R ^y or -SO2W ⁵ ; and W ⁵ is a carbocycle or a heterocycle, wherein W ⁵ is independently substituted with 0 to 3 R ^y groups.

W ⁵ carbocycles and W ⁵ heterocycles may be independently substituted with 0 to 3 R ^y groups. W ⁵ may be a saturated, unsaturated or aromatic ring containing a mono- or bicyclic carbocycle or heterocycle. W ⁵ may have from 3 to 10 ring atoms, for example from 3 to 7 ring atoms. W ⁵ rings are saturated rings that contain 3 ring atoms, saturated or monounsaturated, which contain 4 ring atoms, saturated or mono- or diunsaturated, which contain 5 ring atoms, and saturated, mono- or diunsaturated, or aromatic, which contain 6 ring atoms.

The W ⁵ heterocycle may be a monocycle having 3 to 7 ring members (2 to 6 carbon atoms and 1 to 3 heteroatoms selected from N, O, P and S) or a bicycle having 7 to 10 ring members (4 to 9 carbon atoms and 1 to 3 heteroatoms selected from N, O, P and S). W ⁵ heterocyclic monocycles may have 3 to 6 ring atoms (2 to 5 carbon atoms and 1 to 2 heteroatoms selected from N, O and S), or 5 or 6 ring atoms (3 to 5 carbon atoms and 1 to 2 heteroatoms selected from N and S). W ⁵ heterocyclic bicycles have 7 to 10 ring atoms (6 to 9 carbon atoms and 1 to 2 heteroatoms selected from N, O and S) arranged in a bicyclo [4,5], [5,5], [5 ,6] or [6,6] system, or 9 to 10 ring atoms (8 to 9 carbon atoms and 1 to 2 heteroatoms selected from N and S) arranged in a bicyclo [5,6] or [6, 6] system. The W ⁵ heterocycle can be linked to Y ² through a carbon, nitrogen, sulfur or other atom by a stable covalent bond.

W ⁵ heterocycles include, for example, pyridyl, dihydropyridinyl isomers, piperidine, pyridazinyl, pyrimidinyl, pyrazinyl, s-triazinyl, oxazolyl, imidazolyl, thiazolyl, isoxazolyl, pyrazolyl, isothiazolyl, furanyl, thiofuranyl, thienyl and pyrrolyl. W ⁵ also includes, but is not limited to, examples such as

- 31 046452

The W ⁵ carbocycles and heterocycles may be independently substituted with 0 to 3 R groups as indicated above. For example, substituted W ⁵ carbocycles include

Examples of substituted phenyl carbocycles include

Embodiments of R ⁷ or R ⁷ together with R ⁴ include structures

- 32 046452 where a represents the point of attachment to R ⁷ ;

b represents the point of attachment to R ⁴ ;

Ar represents phenyl or naphthyl, where phenyl and naphthyl are optionally substituted with 1-3 R ²⁰ groups;

each R ^y is, independently, (C ₁ -C ₈ )αalkyl or ^-^-carbocyclyl, where alkyl and carbocyclyl are optionally substituted with 1-3 R ²⁰ groups;

each R is, independently, H, (C ₁ -C ₆ )αalkyl or arylalkyl; and each R ²⁰ is, independently, halogen, CN, N(R) ₂ , OR, -SR, -S(O)R, -S(O) ₂ R, -S(O)(OR), -S (O)2(OR), -C(=O)R, -C(=O)OR or C(=O)N(R)2.

Embodiments

W ² compounds of formulas I-III and formulas IV-VI include substructures such as

where each Y ^2b is, independently, O or N(R). In another aspect of this embodiment, each Y ^2b is O and each R ^x is independently

wherein M12c is 1, 2 or 3, and each Y ² is, independently, a bond, O, CR ₂ or S. In another aspect of this embodiment, one Y ^2b -R ^x is NH(R), and the other Y ^2b -R ^x represents OR ^x , where R ^x represents

CR ₃

M12c where M12c represents 2. In another aspect of this, each R ^x represents an independent embodiment of each Y ^2b represents a co-

where M12c represents 2. In another aspect of this, each R ^x represents an independent embodiment of each Y ^2b represents a co-

where M12c represents 1 and Y ² represents a bond, O or CR ₂ . Other connection embodiments

formulas I-III and formulas IV-VI include substructures such as

- 33 046452

where each Y ³ is, independently, O or N(R). In another aspect of this embodiment, each Y ³ is an O. In another aspect of this embodiment, the substructure is

where R ^y represents W ⁵ as defined here. Another incarnation

formulas I-III and formulas IV-VI includes substructures

where each Y ^2c is, independently, O, N(R ^y ) or S. Another embodiment

compounds of formulas I-III and formulas IV-VI includes substructures where one W ¹ or W ² , together with either R ³ or R ⁴ represents -Y ³ -, and the other W ¹ or W ² represents formula Ia. Such an embodiment is a compound of formula Ib selected from

- 34 046452

In another aspect of an embodiment of Formula Ib, Y and Y ³ are each O.

In another aspect of formula Ib, W ¹ or W ² is Y ^2b -R ^x ; Y, Y ³ and Y ^2b each represent O, and R ^x represents

wherein M12c is 1, 2 or 3, and each Y ² is, independently, a bond, O, CR ₂ or S. In another aspect of the embodiment of formula Ib, W ¹ or W ² is Y ^2b -R ^x ; Y, Y ³ and Y ^2b each represent O, and R ^x represents

wherein M12c is 2. In another aspect of formula Ib, W ¹ or W ² is Y ^2b -R ^x ; Y, Y ³ and Y ^2b each represent O, and R ^x represents

- 35 046452

where M12c represents 1 and Y ² represents a bond, O or CR ₂ . Another incarnation

compounds of formulas I-III and formulas IV-VI includes the substructure

where W ⁵ represents a carbocycle such as phenyl or substituted phenyl. In another aspect of this embodiment, the substructure is

O where Y ^2b represents O or N(R), and the phenyl carbocycle is substituted with 0 to 3 R groups. In another aspect of this embodiment of the substructure R ^x is

where M12c represents 1, 2 or 3, and each Y ² represents, independently, a bond, O, CR2 or S.

Another incarnation

formulas I-III and formulas IV-VI includes the substructure

The amino acid's chiral carbon and lactate moieties can be either R- or S-configuration, or a racemic mixture.

Another incarnation

- 36 046452

formulas I-III and formulas IV-VI represents the substructure

where each Y ² represents, independently, -O- or -NH-. In another aspect of this embodiment, R ^y is (C1-C8)alkyl, (C1-C8)substituted alkyl, (C2-C8)alkenyl, (C2-C8)substituted alkenyl, (C2-C8)alkynyl, or (C2-C8) )substituted alkynyl. In another aspect of this embodiment, R ^y is (C1-C8)alkyl, (C1- _C8 )substituted alkyl, ( _C2 - _C8 )alkenyl, ( _C2 - _C8 )substituted alkenyl, ( _C2C8 )alkynyl, or (C ₂ -C ₈ )substituted alkynyl; and R is CH3. In another aspect of this embodiment, R ^y is (C1- _C8 )alkyl, (C1- _C8 )substituted alkyl, ( _C2 - _C8 )alkenyl, ( _C2C8 )substituted alkenyl, ( _C2 - _C8 )alkynyl or (C ₂ -C ₈ )substituted alkynyl; R is CH3; and each Y ² represents -NH-. In an aspect of this embodiment, W ¹ and W ² are, independently, nitrogen-linked, naturally occurring amino acids or naturally occurring amino acid esters. In another aspect of this embodiment, W ¹ and W ² are, independently, naturally occurring 2-hydroxy carboxylic acids or naturally occurring 2-hydroxy esters of carboxylic acids, wherein the acid or ester is linked to P via a 2-hydroxy group.

Another incarnation

formula I, formula II, formula III, formula IV, formula V or formula VI is the structure

In one embodiment, each R ^x is, independently, (C1-C8)αalkyl. In another aspect of this embodiment, each R ^x is, independently, C6- _C20 aryl or C6-C20 substituted aryl.

In a preferred embodiment

selected from

- 37 046452

Another incarnation

formulas I-III and formulas IV-VI represents the substructure

where W ¹ and W ² are independently selected from one of the formulas in the table. 20.1-20.37 and table. 30.1 below. Variables used in the table. 20.1-20.37 (for example, W ²³ , R ²¹ , etc.), refer only to table. 20.1-20.37, unless otherwise stated.

Variables used in the table. from 20.1 to 20.37 have the following definitions:

each R ²¹ is, independently, H or (C1-C ₈ )alkyl;

each R ²² is, independently, H, R ²¹ , R ²³ or R ²⁴ , where each R ²⁴ is, independently, substituted with 0 to 3 R ²³ ;

each R ²³ is, independently, R ^23a , R ^23b , R ^23c or R ^23d , with the proviso that when R ²³ is bonded to a heteroatom, then R ²³ is R ^23c or R ^23d ;

each R ^23a is, independently, F, Cl, Br, I, -CN, N ₃ or -NO ₂ ;

each R ^23b represents, independently, Y ²¹ ;

each R ^23c is, independently, -R ^2x , -N(R ^2x )(R ^2x ), -SR ^2x , -S(O)R ^2x , -S(O)2R ^2x , -S(O)(OR ^2x ), -S(O)2(OR ^2x ), -OC(=Y ²¹ )R ^2x , -OC(=Y ²¹ )OR ^2x , -OC(=Y ²¹ )(N(R ^2x )(R ^2x )), -SC(=Y ²¹ )R ^2x , -SC(=Y ²¹ )OR ^2x , -SC(=Y ²¹ )(N(R ^2x )(R ^2x )), -N(R ^2x )C( =Y ²¹ )R ^2x , -N(R ^2x )C(=Y ²¹ )OR ^2x or

-N(R ^2x )C(=Y ²¹ )(N(R ^2x )(R ^2x ));

each R ^23d is, independently, -C(=Y ²¹ )R ^2x , -C(=Y ²¹ )OR ^2x or -C(=Y ²¹ )(N(R ^2x )(R ^2x ));

each R ^2x is, independently, H, (C1-C8)alkyl, (C2-C8)alkenyl, (C2-C8)alkynyl, aryl, heteroaryl; or two R ^2x taken together with the nitrogen to which they are both attached form a 3-7 membered heterocycle, wherein any carbon atom of said heterocyclic ring may optionally be replaced by -O-, -S- or -NR ²¹ -; and wherein one or more non-terminal carbon atoms of each said (C1-C8)alkyl may optionally be replaced by -O-, -S- or -NR ²¹ -;

each R ²⁴ is, independently, (C1- _C8 )alkyl, ( _C2 - _C8 )alkenyl or ( _C2 - _C8 )alkynyl;

each R ²⁵ is, independently, R ²⁴ wherein each R ²⁴ is substituted with 0 to 3 R ²³ groups;

each R ^25a is, independently, (C1- _C8 )alkylene, ( _C2 - _C8 )alkenylene or ( _C2 - _C8 )alkynylene, each of the above (C1- _C8 )alkylene, ( _C2 -C ₈ ) alkenylene or (C ₂ -C ₈ ) alkynylene is substituted with 0-3 R ²³ groups;

each W ²³ is, independently, W ²⁴ or W ²⁵ ;

each W ²⁴ is, independently, R ²⁵ , -C(=Y ²¹ )R ²⁵ , -C(=Y ²¹ )W ²⁵ , -SO2R ²⁵ or -SO2W ²⁵ ;

each W ²⁵ is, independently, a carbocycle or a heterocycle, wherein W ²⁵ is independently substituted with 0 to 3 R ²² groups; And

- 38 046452 each Y ²¹ represents, independently, O or S.

Table 20.1

Table 20.2

Table 20.3

- 39 046452

Table 20.4

CH ₃ Mo 11 o 20 ( NzM ^? '^o^ ^x y°''w ²³ o 23 NzM A R ²¹ O 26 NzM Mmo 29 n ₃ s. Mmo 31 N ₃ C 0 СНз /СН ₃ /Ск ,сн ₃ '-М 3 \ ₀ ^\м хз О сн ₃ 21 СН ₃ СН ₃ JL 3 Y СН ₃ О 22 Table 20.5 НзМ Н ₃ с. 'Chu'u ⁰ ''^ ²⁵ ^ΆγΜ^Μρ ²⁴ o o 24 25 n ₃ s. n ₃ s %gu°^n ^g ^сгу°\ _С n ₃ о о 27 28 НзМ / ^{сн зг} \о mm ^ _СНз О 30 Table 20.6 Н ₃ С. /СН ₃ /О. /CH ₃ ' Μγγ O sn ₃ 32 A CH ₃ ' D ^/O mmMSn ₃ o 33

- 40 046452

Table 20.7

Table 20.8

Table 20.9

- 41 046452

Table 20.10

Table 20.11

Table 20.12

- 42 046452

Table 20.13

Table 20.14

Table 20.15

- 43 046452

Table 20.16

Table 20.17

Table 20.18

Table 20.19

- 44 046452

Table 20.20

Table 20.22

Table 20.23

Table 20.21

Table 20.24

- 45 046452

Table 20.25

Table 20.26

Table 20.28

Table 20.27

- 46 046452

Table 20.29

Table 20.30

- 47 046452

Table 20.31

200

201

ABOUT

202

203

O Y ,R^ ^a JL /H . , o o o *\ J o 204 o o o sn ₃ 206 Table 20.32 O |^^CH ₃ ^g \ /R\ ^a JL Y Ύ O O O x 208 O CH ₃ L _R 25a I J< ^CH3 / O o o sn ₃ o 210 in Their << R?? ^a bx x ° o o o 212 O _R 25 _a II _C |_| ^{213 ​} _{​ ​ ​} ^​ _{​ ​} ^{​ ​}

- 48 046452

Table 20.33

Table 20.34

Table 20.35

- 49 046452

Table 20.36

Table 20.37

Table 30.1

- 50 046452

Embodiments of R ^x include esters, carbamates, carbonates, thioesters, amides, thioamides and urea groups

Any reference to compounds of the invention described herein also includes references to physiologically acceptable salts. Examples of physiologically acceptable salts of the compounds of the invention include salts derived from corresponding bases such as alkali metals or alkaline earths (eg, Na ⁺ , Li ⁺ , K ⁺ , Ca ⁺² and Mg ⁺² ), ammonium and NR4 ⁺ (where R is defined Here). Physiologically acceptable salts of the nitrogen atom or amino group include: (a) acid addition salts formed with inorganic acids, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid and the like; (b) salts formed with organic acids, such as, for example, acetic acid, oxalic acid, tartaric acid, succinic acid, maleic acid, fumaric acid, gluconic acid, citric acid, malic acid, ascorbic acid, benzoic acid, isethionic acid , lactobionic acid, tannic acid, palmitic acid, alginic acid, polyglutamic acid, naphthalenesulfonic acid, methanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, naphthalenedisulfonic acid, polygalacturonic acid, malonic acid, sulfosalicylic acid, glycolic acid, toat, pamoate, salicylic acid, stearic acid, phthalic acid, mandelic acid, lactic acid, ethanesulfonic acid, lysine, arginine, glutamic acid, glycine, serine, threonine, alanine, isoleucine, leucine, etc.; and (c) salts formed from elemental anions, for example, chlorine, bromine and iodine. Physiologically acceptable salts of a hydroxyl group compound include an anion of the compound in combination with a suitable cation such as Na ⁺ and NR ₄ ⁺ .

For therapeutic use, salts of the active ingredients of the compounds of the invention will be physiologically acceptable, that is, they will be salts derived from a physiologically acceptable acid or base. However, salts of acids and bases that are not physiologically acceptable may also find use, for example, in the preparation or purification of a physiologically acceptable compound. All salts, whether or not formed, formed with physiologically acceptable acids or bases are within the scope of the present invention.

Finally, it should be understood that the compositions include the compounds of the invention in their non-ionized as well as zwitterionic forms and combinations with stoichiometric amounts of water, as in hydrates.

The compounds of the invention, exemplified by formulas I-III and formulas IV-VI, may have chiral centers, for example, chiral carbon or phosphorus atoms. The compounds of the invention thus include racemic mixtures of all stereoisomers, including enantiomers, diastereomers and atropisomers. In addition, the compounds of the invention include enriched or resolved optical isomers of any or all asymmetric chiral atoms. In other words, chiral centers are obvious from the description and are presented as chiral isomers or racemic mixtures. Both racemic and diastereomeric mixtures, as well as individual optical isomers isolated or synthesized substantially free from their enantiomeric or diastereomeric partners, are within the scope of the invention. Racemic mixtures are separated into individual, essentially optically pure isomers by well-known methods, such as, for example, the separation of diastereomeric salts formed with optically active additives, for example, acids or bases, followed by conversion back to optically active substances. In most cases, the desired optical isomer is synthesized using a stereospecific reaction, starting from the corresponding stereoisomer of the desired starting material.

The term chiral refers to molecules that have the property of not being mirror-imposed on their partner, while the term achiral refers to molecules that are mirror-imposed on their partner.

The term stereoisomers refers to compounds that have the same chemical composition but differ due to the arrangement of atoms or groups in space.

A diastereomer refers to a stereoisomer with two or more centers of chirality and whose molecules are not mirror images of each other. Diastereomers have different physical properties, such as melting point, boiling point, spectral properties and reactivity. Mixtures of diastereomers can be separated by high-resolution analytical procedures such as electrophoresis and chromatography.

Enantiomers refer to two stereoisomeric compounds that are not superimposed mirror images of each other.

- 51 046452

Stereochemical definitions and conventions used here generally follow S. P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984), McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., Stereochemistry of Organic Compounds (1994), John Wiley & Sons, Inc., New York. Many organic compounds exist in optically active forms, i.e. they have the ability to rotate the plane of polarized light. When describing an optically active compound, the prefixes D and L or R and S are used to indicate the absolute configuration of the molecule relative to its chiral center(s). The prefixes d and l, D and L, or (+) and (-) are used to indicate the sign of rotation of the plane of polarized light in conjunction with S, (-), or 1 means that the compound is left-handed, while a compound with the prefix R , (+), or d, is dextrorotatory. For a given chemical structure, these stereoisomers are identical except that they are mirror images of each other. A specific stereoisomer may also be called an enantiomer, and mixtures of such isomers are often called enantiomeric mixtures. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, which can occur where there was no stereoselectivity or stereospecificity in a chemical reaction or process. The terms racemic mixture and racemate refer to an equimolar mixture of two enantiomeric species lacking optical activity.

Whenever a compound described herein is replaced by more than one of the same designated group, for example, R or R ¹ , it will be understood that the groups may be the same or different, i.e. each group is independently selected. Wavy lines, l-wwv indicate the location of the covalent bond attachment to adjacent substructures, groups, fragments or atoms.

The compounds of the invention may also exist as tautomeric isomers in some cases. Although only one delocalized resonant structure can be depicted, all such shapes are contemplated by the invention. For example, en-amine tautomers may exist for purine, pyrimidine, imidazole, guanidine, amidine and tetrazole systems, and all possible tautomeric forms thereof are included within the scope of the invention.

Those skilled in the art will appreciate that pyrrolo[1,2-1][1,2,4]triazine, imidazo[1,5-1][1,2,4]triazine, imidazo[1,2-1 ][1,2,4]triazine and [1,2,4]triazole[4,3-1][1,2,4]triazine nucleosides can exist in tautomeric forms. Thus, for example, but not by way of limitation, structures (a) and (b) may have equivalent tautomeric forms, as shown below

IT

All possible tautomeric forms of heterocycles in all embodiments described herein are within the scope of the invention.

Methods for inhibiting HCV polymerase.

Another aspect of the invention relates to methods of inhibiting HCV polymerase activity, comprising the step of treating a sample suspected of containing HCV with a composition of the invention.

The compositions of the invention may act as inhibitors of HCV polymerase, as intermediates for such inhibitors, or have other utilities as described below. Inhibitors will bind to a site on the surface or cavity of the HCV polymerase that has a geometry unique to the HCV polymerase. Compositions that bind HCV polymerase can bind with varying degrees of reversibility. Those compounds for which binding is essentially irreversible are

- 52 046452 are ideal candidates for use in this method of the invention. Once noted, substantially irreversibly binding compositions are useful as probes for the detection of HCV polymerase. Thus, the invention relates to methods for detecting HCV polymerase in a sample suspected of containing HCV polymerase, comprising the steps of: reacting a sample suspected of containing HCV polymerase with a composition containing a compound of the invention associated with a label; and observing the effect of the sample on the activity of the label. Suitable labels are well known in the diagnostic field and include stable free radicals, fluorophores, radioisotopes, enzymes, chemiluminescent groups and chromogens. The compounds here are labeled in the usual manner using functional groups such as hydroxyl, carboxyl, sulfhydryl or amino acid.

In the context of the present invention, samples suspected of containing HCV polymerases include natural or artificial materials such as living organisms, tissues or cell cultures; biological samples, such as biological material samples (blood, serum, urine, cerebrospinal fluid, tears, sputum, saliva, tissue samples, etc.); laboratory samples; samples of food, water or air; samples of a biological product, such as an extract of cells, in particular, cells that recombinantly synthesize the desired glycoprotein, and the like. Typically, the sample will be expected to contain an organism that produces HCV polymerase, often a pathogenic organism such as HCV. Samples can be maintained in any medium, including water and organic solvent/water mixtures. Samples include living organisms such as humans and artificial materials such as cell cultures.

The processing step of the invention includes adding a composition of the invention to a sample or includes adding a precursor of the composition to a sample. The additional step includes any route of administration as described above.

If desired, HCV polymerase activity after application of the composition can be observed by any method, including direct and indirect methods of detecting HCV polymerase activity. Quantitative, qualitative and semi-quantitative methods for determining HCV polymerase activity are all covered. Typically, one of the screening methods described above is used, however, any other methods, such as observing the physiological properties of a living organism, are also applicable.

Organisms that contain HCV polymerase include the HCV virus. The compounds of the present invention are useful in the treatment or prevention of HCV infection in animals or humans.

However, in screening for compounds capable of inhibiting human immunodeficiency viruses, it should be kept in mind that the results of enzyme assays may not correlate with cell culture assays. Thus, cell-based assays should be the primary screening tool.

Screening for HCV polymerase inhibitors.

The compositions of the invention are tested for HCV polymerase inhibitory activity by any of the conventional methods for assessing enzyme activity. In the context of the present invention, generally, compositions are first tested for inhibition of HCV polymerase in vitro, and compositions showing inhibitory activity are then screened for in vivo activity. Compositions having in vitro Ki (inhibitory constants) of less than about 5 x 10 ^-6 M, generally less than 1 x 10 ^-7 M, and preferably less than about 5 x 10 ^-8 M are preferred for use in vivo.

Useful in vitro tests are described in detail and will not be discussed here. However, the examples describe suitable in vitro assays.

Pharmaceutical compositions.

The compounds of this invention are combined with conventional carriers and excipients, which will be selected in accordance with standard practice. The tablets will contain excipients, glidants, fillers, binders and the like. Aqueous preparations are prepared in sterile form and, when intended for delivery other than oral administration, will generally be isotonic. All compositions will additionally contain excipients such as those set forth in the Handbook of Pharmaceutical Excipients (1986). Excipients include ascorbic acid and other antioxidants, chelating agents such as EDTA, carbohydrates such as dextran, hydroxyalkylcellulose, hydroxyalkylmethylcellulose, stearic acid and the like. The pH of the formulations ranges from 3 to 11, but usually from 7 to 10.

Although this is possible for active ingredients that will only be used, it may be preferable to present them as pharmaceuticals. The compositions for both veterinary and human use of the invention contain at least one active ingredient as defined above, together with one or more acceptable carriers, and therefore, optionally, other therapeutic ingredients. The carrier(s) must be acceptable in terms of compatibility with the other ingredients of the composition and physiologically without

- 53 046452 harmful to the recipient.

Compositions include those suitable for the above routes of administration. The compositions may be presented in unit dosage form and may be prepared by any of the methods well known in the pharmaceutical field. Methods and compositions are generally available from Remington's Pharmaceutical Sciences (Mack Publishing Co., Easton, PA). Such methods include the step of mixing the active ingredient with a carrier that constitutes one or more additional ingredients. In general, compositions are prepared by uniformly and thoroughly mixing the active ingredient with liquid carriers or finely divided solid carriers, or both, and then, if necessary, molding the product.

The compositions of the present invention suitable for oral administration may be presented in the form of discrete units such as capsules, wafers or tablets, each containing a specified amount of the active ingredient in the form of a powder or granules, in the form of a solution or suspension in an aqueous or non-aqueous liquid, or as an oil-in-water liquid or a water-in-oil liquid emulsion. The active ingredient may also be administered as a bolus, porridge or paste.

A tablet is made by compression or molding, optionally with one or more additional ingredients. Compressed tablets can be prepared by compressing in a suitable machine the active ingredient in bulk form, such as powder or granules, optionally mixed with a binder, lubricant, inert diluent, preservative, surfactant or dispersant. Molded tablets can be prepared by molding in a suitable machine a mixture of powdered active ingredient moistened with an inert liquid diluent. Tablets may be coated or uncoated and, optionally, formed in such a way as to provide a slow or controlled release of the active ingredient therefrom.

For infections of the eyes or other external tissues, such as the mouth and skin, the compositions are preferably applied as an ointment or cream containing the active ingredient(s) in an amount of, for example, 0.075 to 20% w/w. (including active ingredient(s) ranging from 0.1 to 20% in increments of 0.1% w/w, such as 0.6% w/w, 0.7% w/w . etc.), preferably from 0.2 to 15% w/w, and most preferably from 0.5 to 10% w/w. When the preparation is in ointment form, the active ingredients can be used with either a paraffin or water-miscible ointment. Alternatively, the active ingredients can be formulated into an oil-in-water cream base.

If desired, the aqueous phase of the cream may include, for example, at least 30% w/w. polyhydric alcohol, that is, an alcohol having two or more hydroxyl groups, such as propylene glycol, butane-1,3-diol, mannitol, sorbitol, glycerin and polyethylene glycol (including PEG 400), and mixtures thereof. Compositions for topical use may desirably include a compound that enhances the absorption or penetration of active ingredients through the skin or other affected areas. Examples of such dermal penetration enhancers include dimethyl sulfoxide and related analogs.

The oil phase of the emulsions of the present invention can be formulated from known ingredients in a known manner. While the phase may contain only an emulsifier (otherwise known as an emulsifier), it desirably contains a mixture of at least one emulsifier with a fat or oil, or both fat and oil. Preferably, the hydrophilic emulsifier is included together with a lipophilic emulsifier which acts as a stabilizer. It is also preferable that it includes both oil and fat. Together, the emulsifier(s) with or without stabilizer(s) constitute the so-called emulsifying wax, and the wax together with the oil and fat constitute the so-called emulsifying ointment, which forms the fatty dispersed phase of cream compositions.

Emulsifiers and emulsion stabilizers suitable for use in the preparation of the formulations of the invention include Tween®60, Span®80, cetostearyl alcohol, benzyl alcohol, myristyl alcohol, glycerol monostearate and sodium lauryl sulfate.

The selection of suitable oils or fats for the preparation is based on achieving the desired cosmetic properties. The cream should preferably be a non-greasy, non-staining and washable product with a suitable consistency to avoid leakage from tubes or other containers. Straight or branched chain esters, mono- or dibasic, such as diisoadipate, isocetyl stearate, propylene glycol diester of coconut fatty acids, isopropyl myristate, decyl oleate, isopropyl palmitate, butyl stearate, 2-ethylhexyl palmitate or a mixture of branched esters known as Crodamol CAP can be used, the last three are the preferred esters. They can be used alone or in combination, depending on the required properties. Alternatively, high melting point lipids such as white soft paraffin and/or liquid paraffin or other mineral oils are used.

Pharmaceutical compositions in accordance with the present invention contain the combination of the invention together with one or more pharmaceutically acceptable carriers or excipients and, optionally, other therapeutic agents. Pharmaceutical compositions containing the active ingredient may be in any form suitable for the intended method

- 54 046452 introductions. When used for oral administration, for example, tablets, lozenges, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, syrups or elixirs may be prepared. Compositions intended for oral administration may be prepared by any method known in the art for the production of pharmaceutical compositions, and such compositions may contain one or more agents, including sweeteners, flavorings, colors and preservatives, so as to provide the drug with a pleasant taste. Tablets containing the active ingredient mixed with a non-toxic pharmaceutically acceptable excipient that is suitable for tablet manufacture are acceptable. These excipients may be, for example, inert diluents such as calcium or sodium carbonate, lactose, calcium or sodium phosphate; granulating agents and disintegrants such as corn starch or alginic acid; binding agents such as starch, gelatin or gum acacia; and lubricants such as magnesium stearate, stearic acid or talc. The tablets may be uncoated or may be coated by known methods including microencapsulation to delay disintegration and absorption in the gastrointestinal tract and thereby provide sustained action over a longer period. For example, delay materials such as glycerin or glyceryl distearate, alone or with a wax, can be used.

Compositions for oral administration may also be presented in the form of hard gelatin capsules, where the active ingredient is mixed with an inert solid diluent, such as calcium phosphate or kaolin, or in the form of soft gelatin capsules, where the active ingredient is mixed with water or an oily vehicle, such as peanut oil, petroleum jelly or olive oil.

The aqueous suspensions according to the invention contain active substances in mixture with excipients suitable for the preparation of aqueous suspensions. Such excipients include suspending agents such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylcellulose, methylcellulose, sodium alginate, polyvinylpyrrolidone, tragacanth and gum acacia, and dispersing or wetting agents such as natural phosphatides (eg lecithin), a condensation product of alkylene oxide with fatty acids (eg, polyoxyethylene stearate), condensation product of ethylene oxide with a long chain aliphatic alcohol (eg, heptadecaethyleneoxycetanol), condensation product of ethylene oxide with a partial ester derived from fatty acids, and hexitol anhydride (eg, polyoxyethylene sorbitan monooleate). Aqueous suspensions may also contain one or more preservatives, such as ethyl or n-propyl-p-hydroxy-benzoate, one or more colors, one or more flavoring agents, and one or more sweeteners, such as sucrose or saccharin.

Oily suspensions can be prepared by suspending the active ingredient in a vegetable oil such as peanut oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. Oral suspensions may contain a thickening agent such as beeswax, paraffin wax, or cetyl alcohol. Sweeteners, such as those listed above, as well as flavoring agents may be added to provide acceptable oral administration. These compositions can be preserved by adding an antioxidant such as ascorbic acid.

The soluble powders and granules of the invention, suitable for preparing an aqueous suspension by adding water, contain the active ingredient in admixture with a dispersing or wetting agent, a suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are, for example, those described above. Additional excipients, such as sweeteners, flavors and colors, may also be present.

The pharmaceutical compositions of the invention can also be in the form of an oil-in-emulsion. The oil phase may be a vegetable oil, such as olive oil or peanut oil, a mineral oil, such as liquid paraffin, or mixtures thereof. Suitable emulsifiers include natural gums such as gum acacia and tragacanth, natural phosphatides such as soy lecithin, esters or partial esters derived from fatty acids and hexitol anhydrides such as sorbitan monooleate, and condensation products of these partial esters with ethylene oxide such as polyoxyethylene sorbitan monooleate. The emulsion may also contain sweeteners and flavorings. Syrups and elixirs can be prepared with sweeteners such as glycerin, sorbitol or sucrose. Such compositions may also contain a demulcent, preservative, flavoring or coloring agent.

The pharmaceutical compositions of the invention may be in the form of sterile injectable preparations, such as sterile injectable aqueous or oily suspensions. This suspension can be prepared according to known techniques using suitable dispersing or wetting agents and suspending agents as mentioned above. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, such as a solution in 1,3-butanediol, or prepared as a lyophilized powder. Among acceptable speakers and races

- 55 046452 solvents that can be used are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile fatty oils can be conveniently used as a solvent or suspension medium. For this purpose, any soft fatty oil can be used, including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid can also be used in the preparation of the injection.

The amount of active ingredient that can be combined with the carrier material to form a unit dosage form will vary depending on the person being treated and the particular route of administration. For example, a sustained release composition for oral administration to a human may contain from about 1 to 1000 mg of active ingredient admixed with a suitable and convenient amount of carrier material, which may range from 5 to 95% of the total composition (wt:wt). The pharmaceutical composition must be capable of providing easily measurable amounts for administration. For example, an aqueous solution intended for intravenous infusion may contain from 3 to 500 mcg of active ingredient per milliliter of infusion solution, making a suitable volume of about 30 mL/hour.

Compositions suitable for topical administration to the eye also include eye drops in which the active ingredient is dissolved or suspended in a suitable vehicle, in particular an aqueous vehicle for the active ingredient. The active ingredient is preferably present in such compositions at a concentration of from 0.5 to 20%, preferably from 0.5 to 10%, and in particular about 1.5% w/w.

Compositions suitable for topical administration to the mouth include lozenges containing the active ingredient in a flavored base, typically sucrose and gum acacia or tragacanth; lozenges containing the active ingredient in an inert base such as gelatin and glycerin or sucrose and acacia; and mouth rinses containing the active ingredient in a suitable liquid carrier.

Compositions for rectal administration may be presented in the form of a suppository with a suitable base containing, for example, cocoa butter or salicylate.

Compositions suitable for intrapulmonary or nasal administration have a particle size, for example, in the range of 0.1 to 500 microns, for example, 0.5, 1, 30, 35, etc., which are administered by rapid inhalation through the nasal passage or by inhalation through the mouth, so as to reach the alveolar sacs. Suitable compositions include aqueous or oily solutions of the active ingredient. Compositions suitable for administration as an aerosol or dry powder can be prepared according to conventional methods and can be delivered with other therapeutic agents, such as compounds hitherto used in the treatment or prevention of HCV infection, as described below.

Compositions suitable for vaginal administration may be in the form of vaginal suppositories, tampons, creams, gels, pastes, foams or sprays, formulations containing in addition to the active ingredient such carriers as are known and appropriate in the art.

Compositions suitable for parenteral administration include aqueous and non-aqueous sterile injectable solutions, which may contain antioxidants, buffers, bacteriostatics and solutes that render the compositions isotonic with the blood of the intended recipient, as well as aqueous and non-aqueous sterile suspensions, which may include suspending agents and thickeners.

The compositions are presented in single dose or multi-dose containers, such as sealed ampoules and vials, and can be stored in a freeze-dried (lyophilized) state, requiring only the addition of a sterile liquid carrier, such as water for injection, immediately prior to use. Improvised solution for injections and suspensions is prepared from sterile powders, granules and tablets, as described previously. Preferred are unit dosage units containing a daily dose or daily subdose unit as described above, or an appropriate portion of the active ingredient.

It should be understood that in addition to the ingredients, particularly those mentioned above, the compositions of this invention may include other agents conventional in the art, taking into account the type of drug in use, for example, those suitable for oral administration may include flavoring agents.

The invention further provides veterinary compositions containing at least one active ingredient as defined above, therefore, together with a veterinary carrier.

Veterinary carriers are materials used for the purpose of administering the composition and may be solid, liquid or gaseous materials that are inert and acceptable in veterinary practice and are compatible with the active ingredient. These veterinary compositions can be administered orally, parenterally, or any other desired route.

The compounds of the invention are used to provide controlled release of a pharmaceutical preparation containing as an active ingredient one or more compounds of the invention (controlled release), in which the release of the active ingredient is controlled and adjusted to reduce the frequency of dosing or to improve pharmacology

- 56 046452 netical parameters or toxicity profile of a given active ingredient.

The effective dose of the active ingredient depends at least on the nature of the condition being treated, the toxicity, whether the compound is being used prophylactically (lower doses) or against an active viral infection, the mode of delivery and the pharmaceutical composition, and will be determined by the physician using routine dose escalation studies. This can be expected to be from about 0.0001 to about 100 mg/kg body weight per day; typically from about 0.01 to about 10 mg/kg body weight per day; more typically from about 0.01 to about 5 mg/kg body weight per day, most often from about 0.05 to about 0.5 mg/kg body weight per day. For example, a suggested daily dose for an adult of approximately 70 kg body weight will range from 1 to 1000 mg, preferably from 5 to 500 mg, and may take the form of one or more doses.

Routes of administration.

One or more compounds of the invention (hereinafter referred to as active ingredients) are administered via any route appropriate to the treatment conditions. Suitable routes include oral, rectal, nasal, topical (including buccal and sublingual), vaginal and parenteral (including subcutaneous, intramuscular, intravenous, intradermal, intrathecal and epidural) and the like. It should be borne in mind that the preferred route may vary depending, for example, on the condition of the recipient. An advantage of the compounds of this invention is that they are orally bioavailable and can be dosed orally.

Combination therapy.

Combinations of compounds of formulas I-III and formulas IV-VI are generally selected depending on the condition being treated, the cross-reactions of the components and the pharmacological properties of the combination. For example, when treating an infection (eg, HCV), the compositions of the invention are combined with other active therapeutic agents (such as those described herein).

The compositions of the invention are also used in combination with one or more other active ingredients. Preferably, the other active ingredients or therapeutic agents are interferons, ribavirin or its analogs, HCV NS3 protease inhibitors, NS5a inhibitors, alpha-glucosidase 1 inhibitors, hepatoprotectors, mevalonate decarboxylase antagonists, renin-angiotensin system antagonists, other anti-fibrotic agents, endothelin antagonists, nucleoside or nucleotide inhibitors of HCV NS5B polymerase, non-nucleoside inhibitors of HCV NS5B polymerase, HCV NS5A inhibitors, TLR-7 agonists, cytophylline inhibitors, HCV IRES inhibitors, pharmacokinetic enhancers or other drugs for the treatment of hepatitis C virus, or mixtures thereof.

In particular, one or more compounds of the present invention may be combined with one or more compounds selected from the group consisting of:

1) interferons, for example, pegylated rGEX-alpha^b (PEG-Intron), pegylated rIFNalpha-33 (Pegasys), rRX-alpha^b (Intron A), rEX-alpha^i (Roferon-A), interferon alpha ( MOR-22, OPC-18, Alfaferone, Alfanative, Multiferon, subalin), interferon alfacon-1 (Infergen), interferon alpha p1 (Wellferon), interferon alpha n3 (Alferon), interferon beta (Avonex, DL-8234 ), interferonomega (omega DUROS, Biomed 510), albinterferon alpha-2b (Albuferon), interferon alpha-XL, BLX883 (Locteron), DA-3021, glycosylated interferon alpha-2b (AVI-005), PEG-Infergen, PEGylated interferon lambda (PEGylated IL-29), and belerofon;

2) ribavirin and its analogues, for example, ribavirin (Rebetol, Copegus) and taribavirin (viramidine);

3) HCV NS3 protease inhibitors, for example, boceprevir (SCH-503034, SCH-7), telaprevir (VX950), VX-813, TMC-435 (TMC435350), ABT-450, BI-201335, BI-1230, MK -7009, SCH-900518, VBY-376, VX-500, GS-9256, GS-9451, BMS-790052, BMS-605339, PHX-1766, AS-101, YH-5258, YH5530, YH5531 and ITMN -191 (R-7227);

4) alpha-glucosidase 1 inhibitors, for example, celgosivir (MX-3253), miglitol and UT-231B;

5) hepatoprotectors, for example, emerikasan (IDN-6556), ME-3738, GS-9450 (LB-84451), silibilin and MitoQ;

6) nucleoside or nucleotide inhibitors of HCV NS5B polymerase, for example, R1626, R7128 (R4048), IDX184, IDX-102, PSI-7851, BCX-4678, valopicitabine (NM-283) and MK-0608;

7) non-nucleoside inhibitors of HCV NS5B polymerase, for example, filibuvir (PF-868554), ABT333, ABT-072, BI-207127, VCH-759, VCH-916, JTK-652, MK-3281, VBY-708, VCH-222 , A848837, ANA598, GL60667, GL59728, A-63890, A-48773, A-48547, BC-2329, VCH-796 (nesbuvir), GSK625433, BILN1941, XTL-2125 and GS-9190;

8) HCVNS5A inhibitors, for example, AZD-2836 (A-831), AZD-7295 (A-689) and BMS-790052;

9) TLR-7 agonists, for example, imiquimod, 852A, GS-9524, ANA-773, ANA-975, AZD-8848 (DSP3025), PF-04878691 and SM-360320;

10) cyclophylline inhibitors, for example DEBIO-025, SCY-635 and NIM811;

11) HCV IRES inhibitors, for example, MCI-067;

12) pharmacokinetic enhancers, for example, BAS-100, SPI-452, PF-4194477, TMC-41629, GS9350, GS-9585 and roxithromycin;

- 57 046452

13) other drugs for the treatment of hepatitis C virus, for example, thymosin alpha 1 (Zadaxin), nitazoxanide (Alinea, NTZ), BIVN-401 (Virostat), PYN-17 (Altirex), KPE02003002, Actilon (CPG10101), GS-9525 , KRN-7000, quoted, GI-5005, XTL-6865, BIT225, PTX-111, ITX2865, TT-033i, ANA 971, NOV-205, tarvacin, EHC-18, VGX-410C, EMZ-702, AVI 4065 , BMS-650032, BMS-791325, Bavituximab, MDX-1106 (ONO-4538), Oglufanide, FK-788 and VX-497 (merimepodib);

14) mevalonate decarboxylase antagonists, for example, statins, HMGCoA synthase inhibitors (for example, himeglusin), squalene synthesis inhibitors (for example, zaragozic acid);

15) angiotensin II receptor antagonists, for example, losartan, irbesartan, olmesartan, candesartan, valsartan, telmisartan, eprosartan;

16) angiotensin-converting enzyme inhibitors, for example, captopril, zofenopril, enalapril, ramipril, quinapril, perindopril, lisinopril, benazepril, fosinopril;

17) other antifibrotic agents, for example, amiloride; And

18) endothelin antagonists, for example, bosentan and ambrisentan.

In yet another embodiment, the present application describes pharmaceutical compositions containing a compound of the present invention, or a pharmaceutically acceptable salt, solvate and/or ester thereof, in combination with at least one additional therapeutic agent and a pharmaceutically acceptable carrier or excipient.

In accordance with the present invention, the therapeutic agent used in combination with the compound or composition of the present invention may be any substance having a therapeutic effect when used in combination with the compound of the present invention. For example, the therapeutic agent used in combination with a compound or composition of the present invention may be interferons, ribavirin or analogs thereof, HCV NS3 protease inhibitors, NS5a inhibitors, alpha-glucosidase 1 inhibitors, hepatoprotectors, mevalonate decarboxylase antagonists, renin-angiotensin system antagonists, others anti-fibrotic agents, endothelin antagonists, nucleoside or nucleotide inhibitors of HCV NS5B polymerase, non-nucleoside inhibitors of HCV NS5B polymerase, HCV NS5A inhibitors, TLR-7 agonists, cytophylline inhibitors, HCV IRES inhibitors, pharmacokinetic enhancers or other compositions for the treatment of hepatitis C virus or their mixtures.

In particular, compositions of one or more compounds of the present invention may be combined with one or more compounds selected from the group consisting of:

1) interferons, for example, PEGylated-rIFN-alpha-2b (PEG-Intron), PEGylated rIFNalpha-3; ·! (Pegasys), rGEX-alpha^b (Intron A), rGEX-alpha^c (Roferon-A), interferon alpha (MOR-22, OPC-18, Alfaferone, Alfanative, Multiferon, subalin), interferon alfacon-1 ( Infergen), interferon alpha-n1 (Wellferon), interferon alpha-i3 (Alferon), interferon-beta (Avonex, DL-8234), interferonomega (omega DUROS, Biomed 510), albinterferon alpha-2b (Albuferon), interferon alpha- XL, BLX883 (Locteron), DA-3021, glycosylated interferon alpha-2b (AVI-005), PEG-Infergen, PEGylated interferon lambda (PEGylated IL-29) and Belerophone;

2) ribavirin and its analogues, for example, ribavirin (Rebetol, Copegus) and taribavirin (viramidine);

3) HCV NS3 protease inhibitors, for example, boceprevir (SCH-503034, SCH-7), telaprevir (VX950), VX-813, TMC-435 (TMC435350), ABT-450, BI-201335, BI-1230, MK -7009, SCH-900518, VBY-376, VX-500, GS-9256, GS-9451, BMS-790052, BMS-605339, PHX-1766, AS-101, YH-5258, YH5530, YH5531 and ITMN-191 (R-7227);

4) alpha-glucosidase 1 inhibitors, for example, celgosivir (MX-3253), miglitol and UT-231B;

5) hepatoprotectors, for example, emerikasan (IDN-6556), ME-3738, GS-9450 (LB-84451), silibilin and MitoQ;

6) nucleoside or nucleotide inhibitors of HCV NS5B polymerase, for example, R1626, R7128 (R4048), IDX184, IDX-102, PSI-7851, BCX-4678, valopicitabine (NM-283) and MK-0608;

7) non-nucleoside inhibitors of HCV NS5B polymerase, for example, filibuvir (PF-868554), ABT333, ABT-072, BI-207127, VCH-759, VCH-916, JTK-652, MK-3281, VBY-708, VCH-222 , A848837, ANA598, GL60667, GL59728, A-63890, A-48773, A-48547, BC-2329, VCH-796 (nesbuvir), GSK625433, BILN1941, XTL-2125 and GS-9190;

8) HCV NS5A inhibitors, for example, AZD-2836 (A-831), AZD-7295 (A-689) and BMS-790052;

9) TLR-7 agonists, for example, imiquimod, 852A, GS-9524, ANA-773, ANA-975, AZD-8848 (DSP3025), PF-04878691 and SM-360320;

10) cyclophylline inhibitors, for example DEBIO-025, SCY-635 and NIM811;

11) HCV IRES inhibitors, for example, MCI-067;

12) pharmacokinetic enhancers, for example, BAS-100, SPI-452, PF-4194477, TMC-41629, GS9350, GS-9585 and roxithromycin;

13) other drugs for the treatment of hepatitis C virus, for example, thymosin alpha 1 (Zadaxin), nitazoxanide (Alinea, NTZ), BIVN-401 (Virostat), PYN-17 (Altirex), KPE02003002, Actilon (CPG10101), GS-9525 , KRN-7000, tsivatsir, GI-5005, XTL-6865, BIT225, PTX-111, ITX2865, TT-033i, ANA 971, NOV-205, tarvacin, EHC-18, VGX-410C, EMZ-702, AVI 4065 , BMS-650032, BMS-791325, Bavituximab,

- 58 046452

MDX-1106 (ONO-4538), Oglufanide, FK-788 and VX-497 (merimepodib);

14) mevalonate decarboxylase antagonists, for example, statins, HMGCoA synthase inhibitors (for example, himeglusin), squalene synthesis inhibitors (for example, zaragozic acid);

15) angiotensin II receptor antagonists, for example, losartan, irbesartan, olmesartan, candesartan, valsartan, telmisartan, eprosartan;

16) angiotensin-converting enzyme inhibitors, for example, captopril, zofenopril, enalapril, ramipril, quinapril, perindopril, lisinopril, benazepril, fosinopril;

17) other anti-fibrotic agents, for example, amiloride; And

18) endothelin antagonists, for example, bosentan and ambrisentan.

In yet another embodiment, the present application provides a combination of a pharmaceutical agent comprising:

a) a first pharmaceutical composition containing a compound of the present invention or a pharmaceutically acceptable salt, solvate or ester thereof; And

b) a second pharmaceutical composition containing at least one additional therapeutic agent selected from the group consisting of HIV protease inhibitory compounds, HIV non-nucleoside reverse transcriptase inhibitors, HIV reverse transcriptase inhibitors, HIV nucleotide reverse transcriptase inhibitors, HIV integrase inhibitors , gp41 inhibitors, CXCR4 inhibitors, gp120 inhibitors, CCR5 receptor inhibitors, interferons, ribavirin analogues, NS3 protease inhibitors, NS5a inhibitors, alpha-glucosidase 1 inhibitors, cyclophilin inhibitors, hepatoprotectors, non-nucleoside inhibitors of hepatitis C virus and other drugs for the treatment of hepatitis C virus , and their combinations.

Combinations of compounds of formulas I-III and formulas IV-VI and additional active therapeutic agents may be selected for the treatment of patients infected with hepatitis C virus and other diseases such as HIV infections. Accordingly, the compounds of formulas I-III and formulas IV-VI can be combined with one or more compounds used in the treatment of HIV, for example, HIV protease inhibitory compounds, HIV non-nucleoside reverse transcriptase inhibitors, HIV reverse transcriptase inhibitors, HIV nucleotide reverse transcriptase inhibitors, HIV integrase inhibitors, gp41 inhibitors, CXCR4 inhibitors, gp120 inhibitors, CCR5 receptor inhibitors, interferons, ribavirin analogues, NS3 protease inhibitors, NS5a inhibitors, alpha-glucosidase 1 inhibitors, cyclophilin inhibitors, hepatoprotectors, non-nucleoside HCV inhibitors, as well as other drugs for HCV treatment.

In particular, one or more compounds of the present invention may be combined with one or more compounds selected from the group consisting of:

1) HIV protease inhibitors, for example, amprenavir, atazanavir, fosamprenavir, indinavir, lopinavir, ritonavir, lopinavir + ritonavir, nelfinavir, saquinavir, tipranavir, brecanavir, darunavir, TMC-126, TMC-114, mosenavir (DMP-450), JE -2147 (AG1776), AG1859, DG35, L-756423, RO0334649, KNI-272, DPC-681, DPC-684, GW640385X, DG17, PPL-100;

2) HIV non-nucleoside reverse transcriptase inhibitors, for example, capravirine, emivirine, delaviridine, efavirenz, nevirapine, (+) calanolide A, etravirine, GW5634, DPC-083, DPC-961, DPC-963, MIV-150 and TMC-120, TMC-278 (rilpivirine), efavirenz, BILR 355 BS, VRX 840773, UK453, 061, RDEA806;

3) HIV nucleoside reverse transcriptase inhibitors, for example, zidovudine, emtricitabine, didanosine, stavudine, zalcitabine, lamivudine, abacavir, amdoxovir, elvucitabine, alovudine, MIV-210, racivir (±-FTC), D-d4FC, emtricitabine, phosphazide, fosivudine tidoxil, fosalvudine tidoxil, apricitibine (AVX754), amdoxovir, KP-1461, abacavir + lamivudine, abacavir + lamivudine + zidovudine, zidovudine + lamivudine;

4) HIV nucleotide reverse transcriptase inhibitors, for example, tenofovir, tenofovir disoproxil fumarate + emtricitabine, tenofovir disoproxil fumarate + emtricitabine + efavirenz and adefovir;

5) HIV integrase inhibitors, for example, curcumin, curcumin derivatives, chicoric acid, chicoric acid derivatives, 3,5-dicaffeoylquinic acid, 3,5dicaffeoylquinic acid derivatives, aurintricarboxylic acid, aurintricarboxylic acid derivatives, caffeic acid phenethyl ester, caffeic phenethyl ester derivatives acid, tyrphostin, tyrphostin derivatives, quercetin, quercetin derivatives, S-1360, zintevir (AR-177), L870812 and L-870810, MK-0518 (raltegravir), BMS-707035, MK-2048, BA-011, BMS- 538158, GSK364735C;

6) gp41 inhibitors, for example, enfuvirtide, sifuvirtide, FB006M, TRI-1144, SPC3, DES6, Locus gp41, CovX and REP 9;

7) CXCR4 inhibitors, for example, AMD-070;

8) introductory inhibitors, for example, SP01A, TNX-355;

9) gp120 inhibitors, for example, BMS-488043 and BlockAide/CR;

10) G6PD and NADH oxidase inhibitor, for example, immunotin;

- 59 046452

10.1) CCR5 receptor inhibitors, for example, aplaviroc, vicriviroc, INCB9471, PRO-140, INCB15050, PF-232798, CCR5mAb004 and maraviroc;

11) interferon, for example, PEGylated rGEX-alpha^b, PEGylated rIFN-alpha-2a, rIFN-alpha-2b, interferon alpha-L XL, gIFN-alpha-2a, interferon alpha-consensus, Infergen, Rebif, Lokteron, AVI -005, PEG-Infergen, PEG-interferon-beta, oral interferon alpha, Feron, reaferon, Intermax alpha, r-IFN-beta, infergen + actimmune, IFN-omega with DUROS and Albuferon;

12) analogues of ribavirin, for example, rebetol, copegus, VX-497, viramidine (taribavirin);

13) NS5a inhibitors, for example, A-831, A-689 and BMS-790052;

14) NS5b polymerase inhibitors, for example, NM-283, valopicitabine, R1626, PSI-6130 (R1656), IDX184, PSI-7851, HCV-796, BILB 1941, MK-0608, NM-107, R7128, VCH-759, PF-868554, GSK625433 and XTL-2125;

15) NS3 protease inhibitors, for example, SCH-503034 (SCH-7), VX-950 (telaprevir), ITMN-191 and BILN-2065;

16) alpha-glucosidase 1 inhibitors, for example, MX-3253 (celgosivir) and UT-231B;

17) hepatoprotectors, for example, IDN-6556, ME 3738, MitoQ and LB-84451;

18) non-nucleoside inhibitors of the hepatitis C virus, for example, benzimidazole derivatives, benzo1,2,4-thiadiazine derivatives and phenylalanine derivatives;

19) other drugs for the treatment of hepatitis C virus, for example, Zadaxin, nitazoxanide (Alinea), BIVN-401 (Virostat), DEBIO -025, VGX-410C, EMZ-702, AVI 4065, bavituximab, oglufanide, PYN17, KPE02003002, actilone (CPG-10101), KRN-7000, Cyvacira, GI-5005, ANA-975, XTL-6865, ANA 971, NOV -205, Tarvacina, EHC-18 and NIM811;

19.1) pharmacokinetic enhancers, for example, BAS-100 and SPI452;

20) RNase H inhibitors, for example, ODN-93 and ODN-112;

21) other anti-HIV agents, for example, VGV-1, PA-457 (bevirimate), Ampligen, HRG214, cytolin, polymune, VGX-410, KD247, AMZ 0026, CYT 99007, A-221 HIV, BAY 50-4798 , MDX010 (iplimumab), PBS119, ALG889 and PA-1050040.

In addition, any of the compounds of the invention may be combined with one or more other active therapeutic agents into a single dosage form for simultaneous or sequential administration to a patient. Combination therapy can be administered as a simultaneous or sequential regimen. When administered sequentially, the combinations may be administered in two or more administrations.

Coadministration of a compound of the invention with one or more other active therapeutic agents generally refers to the simultaneous or sequential administration of a compound of the invention and one or more other active therapeutic agents such that a therapeutically effective amount of the compound of the invention and one or more other active therapeutic agents are both present in the patient's body.

Co-administration involves administering unit doses of compounds of the invention before or after administration of a single dose of one or more other active therapeutic agents, for example, administering a compound of the invention within seconds, minutes or hours of administration of one or more other active therapeutic agents. For example, a unit dose of a compound of the invention may be administered first, followed by a unit dose of one or more other active therapeutic agents over several seconds or minutes. In addition, a dose of one or more other therapeutic agents may be administered first, followed by a single dose of a compound of the invention over several seconds or minutes. In some cases, it may be desirable to administer a unit dose of a compound of the invention first and then, after a period of several hours (eg, 1-12 hours), by administering a unit dose of one or more other active therapeutic agents. In other cases, it may be desirable to administer a dose of one or more other active therapeutic agents first and then, after a period of several hours (eg, 1-12 hours), by administering a single dose of a compound of the invention.

Combination therapy may provide synergy and synergism, i.e. the effect achieved when the active ingredients used together are greater than the sum of the effects resulting from using the compounds separately. A synergistic effect can be achieved when the active ingredients are: (1) formulated and administered together or administered simultaneously in a combination formulation, (2) administered alternately or in parallel as separate formulations, or (3) in some other regimen. When delivered in sequential therapy, a synergistic effect can be achieved when the compounds are administered or delivered sequentially, for example, in separate tablets, pills or capsules, or different injections in separate syringes. In general, during alternation therapy, an effective dose of each active ingredient is administered one after the other, that is, sequentially, while in combination therapy, effective doses of two or more active ingredients are administered together. A synergistic antiviral effect means an antiviral effect that is greater than the predicted purely additive effect of the combination of individual compounds.

In yet another embodiment, the present application provides methods for inhibiting

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HCV polymerases in a cell, comprising: contacting a cell infected with HCV with an effective amount of a compound of Formulas I-III and Formulas IV-VI, or a pharmaceutically acceptable salt, solvate and/or ester thereof, whereby the HCV polymerase is inhibited.

In yet another embodiment, the present application provides methods for inhibiting HCV polymerase in a cell, comprising: contacting an HCV-infected cell with an effective amount of a compound of Formulas I-III and Formulas IV-VI or a pharmaceutically acceptable salt, solvate and/or ester thereof, and with at least one additional active therapeutic agent that inhibits HCV polymerase.

In yet another embodiment, the present application provides methods for inhibiting HCV polymerase in a cell, comprising: contacting an HCV-infected cell with an effective amount of a compound of Formulas I-III and Formulas IV-VI or a pharmaceutically acceptable salt, solvate and/or ester thereof, and at least one additional active therapeutic agent selected from the group consisting of one or more interferons, ribavirin or analogs thereof, HCV NS3 protease inhibitor, NS5a inhibitors, alpha-glucosidase 1 inhibitor, hepatoprotectors, mevalonate decarboxylase antagonist, renin-angiotensin system antagonist, other anti-fibrotic agents, endothelin antagonists, nucleoside or nucleotide inhibitors of HCV NS5B polymerase, non-nucleoside inhibitors of HCV NS5B polymerase, HCV NS5A inhibitors, TLR-7 agonists, cyclophilin inhibitors, HCV IRES inhibitors, pharmacokinetic enhancers and other drugs for the treatment of HCV or mixtures thereof .

In yet another embodiment, the present application provides methods for treating HCV in a patient, comprising administering to the patient a therapeutically effective amount of a compound of Formulas IIII and Formulas IV-VI, or a pharmaceutically acceptable salt, solvate and/or ester thereof.

In yet another embodiment, the present application provides methods for treating HCV in a patient, comprising administering to the patient a therapeutically effective amount of a compound of Formulas IIII and Formulas IV-VI, or a pharmaceutically acceptable salt, solvate and/or ester thereof, and at least one additional active therapeutic agent, which inhibits HCV polymerase.

In yet another embodiment, the present application provides methods for treating HCV in a patient, comprising administering to the patient a therapeutically effective amount of a compound of Formulas IIII and Formulas IV-VI, or a pharmaceutically acceptable salt, solvate and/or ester thereof, and at least one additional active therapeutic agent, selected from the group consisting of one or more interferons, ribavirin or analogues thereof, HCV NS3 protease inhibitor, NS5a inhibitor, alpha-glucosidase 1 inhibitors, hepatoprotectors, mevalonate decarboxylase antagonist, renin-angiotensin system antagonist, other antifibrotic agents, endothelin antagonists, nucleoside or nucleotide HCV NS5B polymerase inhibitors, non-nucleoside HCV NS5B polymerase inhibitors, HCV NS5A inhibitors, TLR-7 agonists, cyclophilin inhibitors, HCV IRES inhibitors, pharmacokinetic enhancers and other drugs for the treatment of HCV, or mixtures thereof.

In yet another embodiment, the present application provides for use of a compound of the present invention, or a pharmaceutically acceptable salt, solvate and/or ester thereof, for the preparation of a medicament for the treatment of HCV infection in a patient.

Metabolites of the compounds of the invention.

Also within the scope of this invention are the in vivo metabolic products of the compounds described herein, to the extent that such products are new and not obvious compared to the prior art. Such products may be the result of, for example, oxidation, reduction, hydrolysis, amidation, esterification, and the like. of the administered compound mainly due to enzymatic processes. Accordingly, the invention includes novel and non-obvious compounds prepared by a process comprising contacting a compound of the invention with a mammal for a period of time sufficient to produce metabolic products. Such products are typically identified by the prepared radiolabel (eg, ¹⁴ C or ³ H) of the compound of the invention when administered parenterally at a dose that is detectable (eg, greater than about 0.5 mg/kg) in animals such as rats, mice , guinea pigs, monkeys, or humans by allowing sufficient time for metabolism to occur (usually about 30 s to 30 h) and isolating the metabolic products from urine, blood, or other biological samples. These products are easy to isolate because they are labeled (others are isolated using antibodies that can bind the surviving epitopes in the metabolite). The structure of the metabolite is determined in the usual manner, for example by MS or NMR analysis. In general, metabolite analysis is performed in the same manner as conventional drug metabolism studies well known to those skilled in the art. Metabolic products, as long as they are not found in vivo, can be used in diagnostic tests for therapeutic dosing of the compounds of the invention, even if they do not have their own inhibitory activity against HCV polymerase.

Recipes and methods for determining the stability of compounds in gastrointestinal secretion surrogates are known. Compounds are defined here as being stable in the gastrointestinal tract when

- 61 046452 less than about 50 mole percent of the protected groups are deprotected in intestinal or gastric juice surrogates when incubated for 1 hour at 37°C. Just because these compounds are stable in the gastrointestinal tract does not mean that they cannot be hydrolyzed in vivo. The prodrugs of the invention will generally be stable in the digestive system, but may be substantially hydrolyzed to the parent drug in the gastrointestinal tract, liver or other metabolic organs, or within cells in general.

Examples.

Some abbreviations and abbreviations are used to describe experimental details. Although most of them should be understandable to a person skilled in the field, table. 1 contains a list of many of these abbreviations and abbreviations.

Table 1

List of abbreviations and abbreviations

Abbreviation Meaning Ac ₂ O Acetic anhydride AIBN 2,2'-azobis (2-methylpropionitrile) VP benzyl VpVg benzyl bromide B.S.A. bis(trimethylsilyl)acetamide BzCl Benzoyl chloride LED carbonyldiimidazole DABCO 1,4-diazabicyclo[2.2.2]octane DBN 1,5-diazabicyclo[4.3.0]non-5-ene DDQ 2,3-dichloro-5,6-dicyan-1,4-benzoquinone DBU 1,5-diazabicyclo[5.4.0]undec-5-ene DCA dichloroacetamide DCC dicyclohexylcarbodiimide DCM dichloromethane DMAP 4-dimethylaminopyridine DME 1,2-dimethoxyethane DMTC1 Dimethoxytrityl chloride DMSO dimethyl sulfoxide DMTr 4, 4'-dimethoxytrityl DMF dimethylformamide EtOAc ethyl acetate ESI Electroaerosol ionization HMDS hexamethyldisilazane HPLC High Pressure Liquid Chromatography LDA Lithium diisopropylamide LRMS Low resolution mass spectrum MCPBA meta-chloroperbenzoic acid- MeCN acetonitrile MeOH methanol MMTC monomethoxytrityl chloride m/z or m/e Mass to charge ratio M.H. Mass plus 1

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MH- Mass minus 1 MsOH Methanesulfonic acid MS or MS Mass spectrum N.B.S. N-bromosuccinimide Ph phenyl rt or r.t. Room temperature TBAF Tetrabutylammonium fluoride TMSC1 chlorotrimethylsilane TMSBr bromotrimethylsilane TMSI iodotrimethylsilane TMSOTf (trimethylsilyl)trifluoromethylsulfonate TEA triethylamine T.B.A. tributylamine TBAP Tributylammonium pyrophosphate TV SCI tert-butyldimethylsilyl chloride TE AB Triethylammonium bicarbonate T.F.A. Trifluoroacetic acid TLC or tic Thin layer chromatography Tr tripheni lmethyl Tol 4-methylbenzoyl Turbo Grignard 1:1 mixture of isopropyl magnesium chloride and lithium chloride δ ppm to field from tetramethylsilane

Receiving connections.

Connection 1.

¢5 р Bub, BF ₃ -Et ₂ O q

Βζ THF Bz ^z

1a 1b

To a suspension of 7-bromo-2,4-bis-methylsulfanyl-imidazo[2,1-f][1,2,4]triazine (prepared in accordance with WO2008116064, 500 mg, 1.72 mmol) in anhydrous THF (5 ml) BuLi (1.6M in hexane, 1.61 ml, 2.41 mmol) was added dropwise at -78°C. The suspension became a red-brown solution within 5 min and then a mixture of 1a (prepared according to WO 200631725, 675 mg, 1.81 mmol) and boron trifluoride etherate (2.40 ml, 1.89 mmol) in THF (5 ml) is added dropwise to the mixture. After stirring for 2 hours at -78°C, saturated NH ₄ Cl was added to quench the reaction. The mixture was diluted with ethyl acetate, the organic layer was washed with brine and concentrated in vacuo. The residue was purified by silica gel column chromatography (EtOAc/hexanes) to give 1b as a rich yellow foam (650 mg, 67%). ¹ H NMR (400 MHz, CDCl3): δ 8.13 (d, 2H), 8.03 (d, 2H), 7.81 (d, 1H), 7.59 (t, 1H), 7.45 (m, 3H), 7.36 (t, 2H), 6.40 (brs, 1H), 6.01 (dd, 1H), 4.78 (m, 2H), 4.60 (dd, 1H), 2.68 (s, 3H), 2.45 (s, 3H), 1.62 (d, 3H ). ¹⁹ F NMR (376 MHz, CDCl ₃ ): δ -167.5. MS=585.1 (M+H+).

_R z° ^F BF ₃ -Et ₂ O _z o F

DZ DZ

CH ₂ CI ₂

1b 1c

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To a solution of 1b (820 mg, 1.40 mmol) in dichloromethane (20 ml) was added boron trifluoride etherate (2 ml) and triethylsilane (2 ml) and stirred at room temperature for 16 hours. Additional boron trifluoride etherate (1 ml) was added ) and triethylsilane (1 ml) and stirred for 7 days. The mixture was diluted with dichloromethane and saturated sodium bicarbonate solution. The organic layer was washed successively with water, saturated ammonium chloride and brine, dried over magnesium sulfate and concentrated. The residue was purified by silica gel column chromatography (EtOAc/hexane) to give 1c (605 mg, 76%). 1H NMR (400 MHz, CDCl ₃ ): δ 8.10 (d, J=7.2 Hz, 2H), 8.00 (d, J=7.2 Hz, 2H), 7.66 (s, 1H), 7.61 (t, J=7.2 Hz , 1H), 7.53 (t, J=7.2 Hz, 1H), 7.46 (t, J=7.2 Hz, 2H), 7.38 (t, J=7.2 Hz, 2H), 5.78 (m, 2H), 4.80 (dd , 1H), 4.68 (m, 1H), 4.60 (dd, 1H), 2.68 (s, 3H), 2.65 (s, 3H), 1.32 (d, 3H). ¹⁹ F NMR (376 MHz, CDCl ₃ ): δ -149.9. MS=569.1 (M+H+).

Compound 1c (635 mg, 1.12 mmol) was placed in a steel bomb reactor. Liquid ammonia (~30 ml) was added and the reactor bomb was tightly sealed. The mixture was stirred at 50°C for 16 hours. After cooling to room temperature, the ammonia was evaporated and the solid residue was dissolved in THF (10 ml) and MeOH (10 ml). Sodium ethoxide (25 wt%, 0.63 ml) was added and stirred at 60°C for 40 min. The mixture was neutralized with acetic acid and concentrated. The residue was purified by RP HPLC to give product 1d (175 mg, 48%). 1H NMR (400 MHz, DMSO- _d6 ): δ 8.21 (brs, 2H), 7.60 (s, 1H), 5.45 (brs, 1H), 5.43 (d, 1H), 4.91 (t, 1H), 3.92 ( m, 1H), 3.76 (m, 2H), 3.57 (m, 1H), 2.44 (s, 3H), 1.09 (d, 3H). ¹⁹ F NMR (376 MHz, DMSO-d ₆ ): δ -153.5. MS=330.1 (M+H+).

To a solution of 1d (175 mg, 0.53 mmol) in dichloromethane (11 ml) was added MCPBA (370 mg, ~1.5 mmol) and stirred at room temperature for 16 hours. The mixture was concentrated to give crude 1e, which was used for the next reaction without further purification. MS=362.0 (M+H+).

1e 1

Compound 1e (obtained from the previous reaction) was placed in a steel bomb reactor. Liquid ammonia (~30 ml) was added and the reactor bomb was tightly sealed. The mixture was stirred at 115°C for 3 days. After cooling to room temperature, the ammonia was evaporated. The solid residue was purified by RP HPLC to give compound 1 (105 mg, 66% in two steps). 1H NMR (400 MHz, D ₂ O): δ 7.31 (s, 1H), 5.43 (d, J=25.2 Hz, 1H), 4.07 (dd, J=9.6, 23.2, 1H), 3.89 (m, 1H) , 3.83 (dd, J=2.4, 12.8 Hz, 1H), 3.67 (dd, J=4.8, 12.8 Hz, 1H), 1.05 (d, J=22.8 Hz, 3H). ¹⁹ F NMR (376 MHz, D ₂ O): δ -153.5. MS=299.2 (M+H+).

Connection 2.

To a solution of compound 1 (82 mg, 0.28 mmol) in water (340 ml) was added adenosine deaminase (A5168 bovine spleen type IX from Sigma-Aldrich, 0.125 units per ml water), and stirred at

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37°C for 4 hours. The mixture was concentrated and purified by RP HPLC to give compound 2 (56 mg, 68%). 1H NMR (400 MHz, D ₂ O): δ 7.35 (s, 1H), 5.46 (d, J=25.2 Hz, 1H), 4.08 (dd, J=9.6, 22.6, 1H), 3.93 (m, 1H) , 3.87 (dd, J= 2.4, 12.8 Hz, 1H), 3.71 (dd, J=4.8, 12.8 Hz, 1H), 1.12 (d, J=23.2 Hz, 3H). ¹⁹ F NMR (376 MHz, D ₂ O): δ -153.4. MS=300.2 (M+H+).

Connection 3.

1a 3b

TMSCl (2, 66 ml, 21 mmol) and stirred at room temperature for 16 hours under argon. After cooling to -78°C, a solution of BuLi (1.6 M, 21 mL, 33 mmol) in hexane was added dropwise. The mixture is stirred for 1 hour at the same temperature. A solution of 1a (prepared according to WO 200631725, 4.46 g, 12 mmol) in THF (10 ml) was then added. After stirring for 2 hours at -78°C, saturated ammonium chloride solution was added to stop the reaction. The mixture is extracted with ethyl acetate. The organic extract is concentrated in vacuo. The residue was purified by silica gel chromatography (ethyl acetate/hexane) to give 3b as a yellow solid (1.6 g, 32%). MS=507.1 (M+H+).

Alternative procedure for compound 3b using 1,2-bis[(chlorodimethyl)silanyl]ethane instead of chlorotrimethylsilane.

BuLi (1.6 M in hexane, 1 .6 ml) at -78°C. After 30 min, a solution of 1,2-bis[(chlorodimethyl)silanyl]ethane (538 mg, 2.4 mmol) in THF (1.2 mL) was added. After 45 min BuLi (1.6 ml) was added. After an additional 30 min, BuLi (1.5 ml) was added. After 30 min, a solution of 1a (610 mg, 1.64 mmol) in THF (2 ml) was added dropwise. The resulting mixture was stirred at -78°C for 2 hours in an argon atmosphere. Acetic acid (0.7 ml) is added dropwise to stop the reaction, followed by the addition of saturated ammonium chloride solution. The mixture is extracted with ethyl acetate. The organic extract is concentrated in vacuo. The residue was purified by silica gel chromatography (ethyl acetate/hexane) to give 3b (320 mg, 40%). The starting compound 1a was also reduced (350 mg) by chromatography.

To a solution of compound 3b (50 mg, 0.1 mmol) and TMSCN (67 μl, 0.5 mmol) in acetonitrile (2.0 ml) at 0°C was added TMSOTf (91 μl, 0.5 mmol). The reaction mixture was stirred at room temperature for 1 hour, then at 65°C for 3 days. The reaction was stopped with saturated NaHCO ₃ at room temperature and diluted with CH ₃ CO ₂ Et. The organic phase was separated, washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated. The residue was purified by RP-HPLC (acetonitrile/water) to give the desired compound 3c (28 mg, 54%). MS=516.1 (M+H+).

Ammonium hydroxide (28% in water, 0.8 mL) was added to a solution of 3c (56 mg, 0.11 mmol) in methanol (1.2 mL) and stirred at room temperature for 16 h. The mixture was concentrated and the residue was purified with using RP HPLC (water/acetonitrile) to give compound 3 (20 mg, 60%). 1H NMR (500 MHz, D ₂ O): δ 7.88 (s, 1H), 7.07 (d, 1H), 6.92 (d, 1H), 4.17 (m, 2H), 4.04 (dd, 1H), 3.87 (dd , 1H), 1.15 (d, 3H).

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MS=308.1 (M+H+).

Connection 4.

To a solution of compound 3b (60 mg, 0.12 mmol) in methanol (0.5 ml) was added ammonium hydroxide (28% in water, 0.5 ml) and stirred at room temperature for 16 hours. The mixture was concentrated and the residue was purified by RP HPLC (water/acetonitrile) to give compound 4 (25 mg, 70%). MS=299.1 (M+H+).

Connection 5.

Compound 3b was converted to compound 5a using a procedure similar to the conversion of 1b to 1c. Compound 5a is then converted to compound 5 using a procedure similar to the conversion of 3 c to 3. 1H NMR (300 MHz, D2O): δ 7.68 (s, 1H), 6.75 (d, J= 4.5 Hz, 1H), 6.65 (d, J=4.5 Hz, 1H), 5.65 (d, J=25.2 Hz, 1H), 3.95 (m, 3H), 3.74 (dd, 1H), 0.98 (d, J=22.8 Hz, 3H). ¹⁹ F NMR (282 MHz, D2O): δ -154.2. MS=283.2 (M+H+).

General procedure for preparing nucleoside triphosphate.

A nucleoside (~20 mg) was placed in a pear-shaped flask (5-15 ml). Trimethylphosphate was added (0.51.0 ml). The solution was cooled in an ice water bath. POCl ₃ (40-45 mg) was added and stirred at 0°C until the reaction was completed (from 1 to 4 hours; the progress of the reaction is monitored by ion exchange HPLC, analytical samples are prepared by taking ~3 μl of the reaction mixture and diluting it with 1.0 M Et ₃ NH2CO ₃ (30-50 µl)). A solution of pyrophosphate-Bu ₃ N (250 mg) and Bu ₃ N (90-105 mg) in acetonitrile or dimethylformamide (11.5 ml) was then added. The mixture was stirred at 0°C for 0.3 to 2.5 hours, and then the reaction was stopped with 1.0 M Et ₃ NH2CO ₃ (~5 ml). The resulting mixture was stirred for an additional 0.5-1 hour while warming to room temperature. The mixture was concentrated to dryness, redissolved in water (4 ml) and purified using ion exchange HPLC. Fractions containing the target product were concentrated to dryness, dissolved in water (~5 ml), concentrated to dryness and again dissolved in water (~5 ml). NaHCO ₃ (30-50 mg) was added and concentrated to dryness. The residue was dissolved in water and concentrated to dryness again. This process is repeated 2-5 times. The residue is then purified by C-18 HPLC to give the desired product as sodium or salt. Additionally, the crude reaction mixture was subjected to C-18 HPLC followed by ion exchange HPLC purification to obtain the desired triethylammonium salt product.

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TP-1 connection.

Compound TP-1 was prepared by a basic method using Compound 2 as starting material. ¹ H NMR (300 MHz, D2O): δ 7.44 (s, 1H), 5.45 (d, J=25.5 Hz, 1H), 4.0-4.4 (m, 4H), 3.05 (m, NCH2CH3), 1.10 (m, NCH2CH3 and 2'-C-CH3). ³ P NMR (121.4 MHz, D2O): δ -9.5 (d, J=22.1 Hz), -11.0 (d, J=19.9 Hz), -23.2 (t, J=23.0 Hz). ¹⁹ F NMR (282 MHz, D ₂ O): δ -153.9.

TP-2 connection.

Compound TP-2 was prepared by a basic method using compound 3 as starting material. ¹ H NMR (300 MHz, D2O): δ 7.82 (s, 1H), 7.03 (d, 1H), 6.90 (d, 1H), 4.1-4.4 (m, 4H), 3.05 (m, NCH2CH3), 1.10 ( m, NCH2CH3 and 2-C-CH ₃ ). ³¹ P NMR (121.4 MHz, D2O): δ -10.7 (d, J=19.5 Hz), -11.3 (d, J=19.8 Hz), -23.1 (t, J=19.8 Hz).

TP-3 connection.

TP-3

Compound TP-3 was prepared by a basic method using compound 5 as starting material. ¹ H NMR (300 MHz, D ₂ O): δ 7.73 (s, 1H), 6.87 (d, 1H), 6.82 (d, 1H), 5.71 (d, J=24.6 Hz, 1H), 4.0-4.4 ( m, 4H), 3.05 (m, NCH2CH3), 1.14 (m, NCH2CH3), 1.00 (d, J=22.8 Hz, 3H, 2'-C-CH ₃ ). ³¹ P NMR (121.4 MHz, D2O): δ -8.1 (d, J=22.1 Hz), -11.1 (d, J=19.9 Hz), -22.7 (t, J=23.0 Hz). ¹⁹ F NMR (282 MHz, D2O): δ -155.6. MS=520.9 (M-H+).

Connection TP-8a.

HO F

TP-8a

Compound TP-8a was prepared by a basic method using compound 8 as starting material. ¹ H NMR (300 MHz, D2O): δ 7.95 (s, 1H), 7.68 (s, 1H), 5.63 (d, J=25.5 Hz, 1H), 4.0-4.4 (m, 4H), 3.05 (m, NCH2CH3), 1.10 (m, NCH2CH3 and 2'-C-CH3). ³¹ P NMR (121.4 MHz, D2O): δ -9.20 (d, J=22.1 Hz), -11.07 (d, J= 19.9 Hz), -23.82 (t, J=23.0 Hz). ¹⁹ F NMR (282 MHz, D ₂ O): δ -155.9. MS=521.6 (M-H+).

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General procedure for preparing a nucleoside prodrug (method A).

To a solution of the nucleoside (0.1 mmol) in trimethylphosphite (1.0 ml) was added N-tetrazole (42 mg, 0.6 mmol), followed by the addition of 2,2-dimethylthiopropionic acid S-(2-{diisopropylamino-[2( 2,2-Dimethylpropionylsulfanyl)ethoxy]phosphanyloxy}ethyl) ester (prepared according to J. Med Chem, 1985, 38, 3941, 90 mg, 0.2 mmol) at 0°C. After stirring for 2 hours, 30% hydrogen peroxide in H ₂ O (140 µl) was added to the mixture. The mixture was then left to warm to room temperature. After 30 min. While stirring, 1M Na ₂ S ₂ O ₃ in H2O (5 ml) was added to stop the reaction. The organic layer was washed with a saturated aqueous solution of Na ₂ CO ₃ (10 mlx2), brine, and concentrated in vacuo. The residue was purified by RP-HPLC (MeCN-H2O gradient) to give prodrug A.

Connection A-1.

A-1

Compound A-1 was prepared by Method A using Compound 1 as the starting material. 1H NMR (400 MHz, CDC^): δ 7.42 (s, 1H), 5.47 (d, J=26.4 Hz, 1H), 4.95 (brs, 2H), 4.59 (m, 2H), 4.35 (m, 1H, 4'-H), 4.18 (m, 2H, 5'-H), 4.10 (m, 4H), 3.13 (m, 4H), 1.24 (d, 3H), 1.22 (s, 9H), 1.19 (d, 9H). ³¹ P NMR (161.9 MHz, CDCl ₃ ): δ -1.26. MS=667.1 (M+H+).

General procedure for preparing a nucleoside prodrug (method B).

Non-limiting examples of monophosphoramidate prodrugs containing the present invention can be prepared in accordance with General Scheme 1.

Scheme 1

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The general procedure involves reacting an ester salt of amino acid 19b, such as a hydrochloric acid salt, with aryl dichlorophosphate 19a in the presence of about two to ten equivalents of a suitable base to give phosphoramidate 19c. Suitable bases include, but are not limited to, imidazoles, pyridines such as lutidine and DMAP, tertiary amines such as triethylamine and DABCO, and substituted amidines such as DBN and DBU. Tertiary amines are particularly preferred. Preferably, the product from each step is used directly in subsequent steps without recrystallization or chromatography. Specific but non-limiting examples 19a, 19b, 19c can be found in WO 2006/121820, which is incorporated herein by reference in its entirety. Nucleoside base 19d reacts with phosphoramidate 19c in the presence of a suitable base. Suitable bases include, but are not limited to, imidazoles, pyridines such as lutidine and DMAP, tertiary amines such as triethylamine and DABCO, and substituted amidines such as DBN and DBU. Product B can be isolated by recrystallization and/or chromatography.

Connection B-1.

Phenyl ethoxyalaninyl phosphorochloridate (124 mg,

0.42 mmol; obtained according to McGuigan et al, J. Med. Chem. 1993, 36, 1048-1052) was added to a mixture of Compound 3 (20 mg, 0.065 mmol) and Nmethylimidazole (42 μl, 0.52 mmol) in anhydrous trimethylphosphate (0.8 ml). The reaction mixture was stirred for 3 hours at room temperature, and then methanol was added to stop the reaction. The methanol solvent was removed under reduced pressure. The residue was purified by reverse phase HPLC followed by silica gel column chromatography (100% ethyl acetate) to give compound B-1 (10 mg, 27%). ³¹ P NMR (121.4 MHz, CDCl ₃ ): δ -3.42, 3.77. MS = 563.0 (M + H ⁺ ), 561.0 (MH ⁺ ).

Connection B-2.

About 3.1 mmol of 4-chlorophenyl 2-propyloxyalanyl phosphorochloridate (prepared according to McGuigan et al, J. Med. Chem. 1993, 36, 1048-1052) is added to the mixture of about 0.5 mmol of compound 3 and about 3.8 mmol N-methylimidazole in approximately 3 ml anhydrous trimethylphosphate. The reaction mixture is stirred for approximately one hour to 24 hours at room temperature and methanol is added to base the reaction. The methanol solvent is removed under reduced pressure. The residue was purified by reverse phase HPLC to give compound B-2.

Connection B-3.

Compound B-3 was prepared using a similar procedure used for compound B-1. ³¹ P NMR (121.4 MHz, CDCl ₃ ): δ -3.50, 3.76. MS=577.2 (M+H+).

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Connection B-4.

AT 4

Compound B-4 was prepared by a similar procedure as used for compound B-1 NMR (162 MHz, CD3OD): δ 2.2. MS=633.4 (M+H+).

Connection B-5.

^31P

Compound B-5 was obtained by a similar procedure used for compound B-1 NMR (162 MHz, CDCl ₃ ): δ 4.15, 4.27. MS=549.3 (M+H+).

Connection B-6.

31P

Compound B-6 was obtained by a similar procedure used for compound B-1 NMR (162 MHz, CDCl ₃ ): δ 3.50, 4.07. MS=613.1 (M+H+).

Connection B-7.

31P

Compound B-7 was prepared by a similar procedure used for compound using compound 5 as the parent nucleoside. ³¹ P MS=538.1 (M+H+).

Connection B-8.

B-1, iNMR (162 MHz, CDCl ₃ ): δ 3.37, 3.97.

Compound B-8 was prepared by a similar procedure used for compound B-1, using compound 5 as the parent nucleoside. ³¹ P NMR (162 MHz, CDCl ₃ ): δ 3.69, 4.39. MS=588.1 (M+H+).

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Alternative procedure for preparing a nucleoside prodrug (Method C).

To a flask containing L-valine hydrochloride ethyl ester (2.5 g, 13.8 mmol, 1 eq.), CH ₂ Cl ₂ (46 ml, 0.3 M) and phenyl dichlorophosphate (2.1 ml, 13.8 mmol, 1 eq.), and then cooled to -10°C. After 10 minutes, TEA (3.8 ml, 13.8 mmol, 1 eq) was slowly added to the reaction mixture over five minutes. The reaction was carried out for an hour before p-nitrophenol (1.9 g, 13.8 mmol, 1 eq.) was added to the reaction mixture followed by the addition of more TEA (3.8 mL, 13.8 mmol, 1 eq.) over five minutes. The reaction mixture was heated and continued for another two hours. The reaction mixture was concentrated in vacuo and dissolved in diethyl ether (200 ml). Insoluble salts were filtered off and the filtrate was concentrated in vacuo. Flash column chromatography was performed using 4/1 Hex/EtOAc to clear oil as C-1a.

1H NMR (400 MHz, CDCl ₃ ): δ 8.21 (s, 2H), 7.41 - 7.20 (m, 7H), 4.22-4.05 (m, 3H), 2.46 (s, 2H), 1.99 (dd, J=23.0 , 20.1 Hz, 2H), 1.68 (s, 1H), 1.20 -1.05 (m, 8H).

³¹ P NMR (162 MHz, CDCl ₃ ): δ -2.79 (dd, J=28.0, 4.2 Hz).

LC MS m/z 422.99 [M+H+].

Connection C-1.

To the flask containing compound 3 (70 mg, 0.23 mmol, 1 eq.), THF (1 ml, 0.2 M) was added and NMP (1 ml, 0.2 M) was cooled to 0°C. Tert-BuMgCl (560 µl, 2.5 eq, 1 M THF) was added slowly and stirred for 5 min before C-1a phenolate (207 mg, 0.46 mmol, 2 eq dissolved in 500 µl THF) was added ). The reaction mixture was heated to 50°C. The reaction was monitored by LCMS. After completion of the reaction, the mixture was concentrated in vacuo and the residue was purified by HPLC to give compound C-1. 1H NMR (400 MHz, CDCl3) δ 7.87 (s, 1H), 7.24-7.10 (m, 4H), 7.03 (t, J=7.2 Hz, 1H), 6.81 (d, J=4.6 Hz, 1H), 6.52 (d, J=4.7 Hz, 1H), 5.61 (s, 2H), 4.46 (dd, J=24.0, 11.4 Hz, 2H), 4.33-4.14 (m, 2h), 4.06 (dt, J=7.2, 4.2 Hz, 2H), 3.82-3.70 (m, 1H), 3.63 (t, J=10.6 Hz, 2H), 1.98 (s, 1H), 1.17 (dd, J=14.8, 7.6 Hz, 3H), 0.82 (dd , J=22.8, 6.8 Hz, 6H).

³¹ P NMR (162 MHz, CDCl ₃ ): d 5.11.

¹⁹ F NMR (376 MHz, CDCl ₃ ): d -152.28.

LC MS m/z 591.21 [M+H+].

Compound C-2a was prepared in a manner similar to that for compound C-1a, but using a methionine ester. 1H NMR (400 MHz, CDCl3) δ 8.19 (s, 2H), 7.44-7.03 (m, 7H), 4.11 (s, 2H), 3.81 (d, J=44.5 Hz, 1H), 2.04 (s, 3H) , 1.61 (s, 2H), 1.21 (d, J=6.1 Hz, 2H), 1.01-0.65 (m, 4H). ³¹ P NMR (162 MHz, CDCl ₃ ) δ -2.00 (d, J=12.9 Hz). LC MS m/z 455.03 [M+H+].

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Connection C-2.

Compound C-2 was prepared in a manner similar to that for compound C-1 using compound 3 and C-2a.

1H NMR (400 MHz, CDCl3) δ 7.96 (d, J=15.8 Hz, 1H), 7.40-7.06 (m, 13H), 6.93 (d, J=6.7 Hz, 1H), 6.70 (s, 1H), 5.98 (s, 1H), 4.54 (dd, J=21.6, 11.7 Hz, 2H), 4.32 (d, J=12.0 Hz, 2H), 4.14 (dt, J=13.0, 6.4 Hz, 4H), 2.44 (d, J=7.5 Hz, 2H), 2.00 (d, J=16.2 Hz, 5H), 1.89 (s, 2H), 1.35-1.13 (m, 7H).

³¹ P NMR (162 MHz, CDCl ₃ ) d 4.12, 3.58.

¹⁹ F NMR (376 MHz, CDCl ₃ ) d -152.28 (s).

LC MS m/z 623.27 [M+H+].

Compound C-3a was prepared in a manner similar to that for compound C-1a, but using tryptophan ester.

1H NMR (400 MHz, CDCl ₃ ) δ 8.18 - 8.03 (m, 3H), 7.29-7.08 (m, 8H), 7.36-6.98 (m, 3H), 4.41-4.11 (m, 1H), 4.15-3.95 ( m 2H), 3.68-3.80 (m, 1H), 3.33-3.04 (m, 2H), 1.06-1.17 (m, 3H).

³¹ P NMR (162 MHz, CDCl ₃ ) d -2.87, -2.99. LC MS m/z 510.03 [M+H+].

Compound C-3.

Compound C-3 was prepared in a manner similar to that of Compound C-1 using Compound 3 and C-3a.

1H NMR (400 MHz, CDCl ₃ ) δ 8.27 (s, 1H), 7.84 (s, 1H), 7.47 (s, 1H), 7.36-6.77 (m, 11H), 6.57 (s, 1H), 4.40-3.96 (m, 6H), 3.20 (s, 4H), 2.60 (s, 1H), 1.30-1.04 (m, 6H).

³¹ P NMR (162 MHz, CDCl ₃ ) d 4.02, 3.75 ¹⁹ F NMR (376 MHz, CDCl ₃ ) d -152.13.

LC MS m/z 678.32 [M+H+].

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Compound C-4a was prepared in a manner similar to that for compound C-1a by substituting phenylalanine ether.

1H NMR (400 MHz, CDCl3) δ 8.15 (t, J=8.7 Hz, 2H), 7.43-7.11 (m, 10H), 7.04 (ddd, J=11.4, 6.7, 2.9 Hz, 2H), 4.32 (ddd, J=15.3, 11.3, 6.1 Hz, 4H), 4.15-3.99 (m, 7H), 3.74 (td, J=11.0, 5.0 Hz, 8H), 3.01 (d, J=5.7 Hz, 2H), 1.17 (td , J=7.1, 5.2 Hz, 2H).

³¹ P NMR (162 MHz, CDCl3) δ -2.97, -2.99.

LC MS m/z 471.03 [M+H+].

Compound C-4.

C-4

Compound C-4 was prepared in a manner similar to that for compound C-1 using compound 3 and C-4a.

1H NMR (400 MHz, CDCl3) δ 7.92 (d, J=13.2 Hz, 1H), 7.46-6.97 (m, 17H), 6.91 (s, 1H), 6.75 (s, 1H), 4.10 (dd, J= 29.6, 19.2 Hz, 8H), 2.97 (s, 3H), 1.32-1.05 (m, 7H).

³¹ P NMR (162 MHz, CDCl ₃ ) d 5.11.

¹⁹ F NMR (376 MHz, CDCl ₃ ) d -152.34 (s).

LC MS m/z 639.24 [M+H+].

C-5a

Compound C-5a was prepared in a manner similar to the example for compound C-1a, but using proline ether.

1H NMR (400 MHz, CDCl ₃ ) δ 8.20 (d, J=7.8 Hz, 2H), 7.45-7.08 (m, 7H), 4.37 (td, J=8.0, 3.8 Hz, 2H), 4.17-3.98 (m , 2H), 3.61-3.34 (m, 2H), 2.21-1.77 (m, 3H), 1.19 (td, J=7.1, 3.8 Hz, 3H).

³¹ P NMR (162 MHz, CDCl3) d -3.92, -3.96.

LC MS m/z 420.98 [M+H+].

Connection C-5.

C-5

Compound C-5 was prepared in a similar manner to Compound C-1 using Compound 3 and C-5a.

1H NMR (400 MHz, CDCl3) δ 7.95 (d, J=4.5 Hz, 1H), 7.39-7.10 (m, 4H), 6.92 (dd, J=16.0, 4.6 Hz, 1H), 6.69 (s, 1H) , 6.03 (bs, 2H), 4.46-4.36 (m, 1H), 4.36-3.96 (m, 4H), 3.37 (d, J=58.9 Hz, 2H), 2.26-1.66 (m, 4H), 1.39-1.12 (m, 8H).

³¹ P NMR (162 MHz, CDCl ₃ ) d 3.47, 2.75.

¹⁹ F NMR (376 MHz, CDCl ₃ ) d -152.36.

LC MS m/z 589.14 [M+H+].

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Compound C-6 was prepared in a similar manner to Compound C-1 using Compound 3 and the sulfone analogue C-1a.

1H NMR (400 MHz, CDCH) δ 7.93 (s, 1H), 7.89 (s, 1H), 7.35-7.01 (m, 5H), 6.93 (d, J=2.8 Hz, 1H), 6.58 (d, J= 2.8 Hz, 1H), 5.79 (bs, 2H), 4.30 (s, 6H), 4.11 (d, J=7.0 Hz, 6H), 3.10-2.84 (m, 3H), 2.75 (s, 3H), 2.54 ( s, 6H), 1.31-1.15 (m, 6H).

³¹ P NMR (162 MHz, CDCl ₃ ) d 3.39, 3.33.

¹⁹ F NMR (376 MHz, CDCl ₃ ) d -152.40.

LC MS m/z 655.24 [M+H+].

Connection PD-A-8b.

NH ₂

t-BuMgCI

NMR/THF

To a solution of compound 8 (200 mg, 0.71 mmol) in THF (1 ml) and NMP (1 ml) under argon at 0°C was added tert-butyl magnesium chloride (1.0 M in THF, 1.06 ml, 1.06 mmol). After 15 min, compound 30d-1 (280 mg, 0.71 mmol) was added as a solution in THF. After 5 minutes, the reaction mixture was warmed to room temperature and stirred for 2 hours. The reaction mixture was cooled to 0°C, stopped with methanol and concentrated. The reaction mixture was purified by silica gel chromatography and then RP HPLC to give PD-A-8b (225 mg, 59%). ^1H NMR (400 MHz, CDCl ₃ ): δ 8.09 (two s, 1H), 7.54 (two s, 1H), 7.31-7.12 (m, 5H), 5.66 (dd, 1H), 4.52-4.45 (m, 2H), 4.19-4.03 (m, 4H), 3.87-3.69 (m, 1H), 1.35-1.15 (m, 9H).

³¹ P NMR (161 MHz, CDCl ₃ ): δ 4.14 (s), 3.55 (s).

LC/MS=539(M+H+).

Retention time: 1.94 min.

LC: Thermo Electron Surveyor HPLC.

MS: Finnigan LCQ Advantage MAX Mass Spectrometer.

Column: Phenomenex Polar RP 30 mm x 4.6 mm.

Solvents: acetonitrile with 0.1% formic acid, water with 0.1% formic acid.

Gradient: 0-0.1 min. 5% ACN, 0.1-1.95 min. 5-100% ACN, 1.95-3.5 min. 100% ACN, 3.5-3.55 min. 100-5% ACN, 3.55-4 min. 5% ACN.

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Getting 30d-l.

Compound 30d-1 was prepared from 30a in a similar manner to 30d-2 by replacing alanine ethyl ether hydrochloride with alanine isopropyl ether hydrochloride.

Compound (S)-PD-A-8c.

(S)-30d-2 Compound 8

The compound (S)-PD-A-8c was made fundamentally similar to PD-A-8b by replacing (S)-30d-2 with 30d-1. 1H NMR (400 MHz, CDCl ₃ ): δ 8.14 (s, 1H), 7.60 (s, 1H), 7.1-7.3 (m, 5H), 5.66 (dd, 1H), 5.02 (m, 1H), 4.50 ( m, 1H), 4.40 (m, 1H), 4.1-4.3 (m, 2H), 3.98 (m, 1H), 3.78 (m, 1H), 3.18 (brs, 1H), 1.15-1.4 (m, 12H) . ³¹ P NMR (161 MHz, CDCl ₃ ): δ 3.70 (s). LC/MS=553 (M+H+).

Receiving (S)-30d-2.

Alanine isopropyl ether hydrochloride (7.95 g, 47.4 mmol) was suspended in dichloromethane (100 ml). Compound 31a (10 g, 47.4 mmol) was added. Triethylamine (13.2 mL, 95 mmol) is then added dropwise over 15 min (internal reaction temperature; -10°C to 3°C). When the reaction was nearly complete (phosphorus NMR), p-nitrophenol (6.29 g, 45.0 mmol) was added as a solid in one portion. Triethylamine (6.28 ml, 45 mmol) was added to the resulting suspension over 15 minutes. The mixture is then warmed to room temperature. When the reaction was complete, MTBE (100 ml) was added. The white precipitate is removed by filtration. The filter cake was washed with MTBE (3x50 ml). The filtrate and washing waters are combined and concentrated. The residue was purified by silica gel column chromatography (0 to 50% ethyl acetate/hexane) to give 30d-2 as a 1:1 diastereomeric mixture (14.1 g, 77%). 1H NMR (300 MHz, CDCl ₃ ): δ 8.22 (2d, 2H), 7.27.4 (m, 7H), 5.0 (m, 1H), 4.09 (m, 1H), 3.96 (m, 1H), 1.39 ( 2d, 3H), 1.22 (m, 6H). MS=409.0 (M+H+), 407.2 (MH ⁺ ).

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Separation of two diastereomers of compound 30d-2.

The two diastereomers were separated by chiral column chromatography under the following conditions.

Column: Chiralpak IC, 2x25 cm.

Solvent system: 70% heptane and 30% isopropanol (IPA).

Flow rate: 6 ml/min.

Loading volume per pass: 1.0 ml.

Sample loading concentration: 150 mg/ml in 70% heptane and 30% IPA.

^-compound 30d-2: retention time 43 min. ³¹ P NMR (162.1 MHz, CDCl ₃ ): δ -2.99 (s).

^-Compound 30d-2: retention time 62 min. ³¹ P NMR (162.1 MHz, CDCl ₃ ): δ -3.02 (s).

In addition, the two diastereomers were separated by crystallization in the following order.

Compound 30d-2 was dissolved in diethyl ether (~10 ml/g). While stirring, hexane was then added until the solution became cloudy. Crystal grains (~10 mg/gram of compound 30d-2) were added to promote crystallization. The resulting suspension was stirred gently for 16 hours, cooled to ~0°C, stirred for a further 2 hours and filtered to collect crystalline material (recovered yield of crystalline material 35%-35%). The crystalline material contains ~95% ^-compound 30d-2 and ~5% ^-compound 30d-2. Recrystallization provides 99% diastereomerically pure ^)-isomer.

The following exemplary PD-A compounds were prepared following general procedures.

Connection PD-A-8d

Connection PD-A-8f

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General procedure for preparing a nucleoside prodrug (method D).

Non-limiting examples of 3'-O-acylated monophosphoramidate prodrugs containing the present invention can be prepared according to General Scheme 2.

Scheme 2

R ⁸

The general procedure involves the reaction of PD-A (R ⁴ =OH) with a carboxylic acid or activated carboxylic acid, such as an acid chloride or anhydride, which is generally known to those skilled in the art (Journal of Medicinal Chemistry, 2006, 49, 6614 and Organic Letters, 2003, 6, 807). When R ⁸ =NH ₂ , protection of the amino group may be necessary. Briefly, to a solution of compound PD-A in acetonitrile (2 ml) add ^^dimethylformamide dimethyl acetal (~1.1 eq.) and stir at room temperature for 1 hour. When protection of the 6-amino group is complete, the mixture is concentrated to dryness. A dehydrating agent such as DCC (~4 eq), acetonitrile and carboxylic acids (~2 eq) are added to the residue. The mixture is stirred at room temperature for 24-48 hours. Water (0.2 ml) and trifluoroacetic acid (0.1 ml) are added at 0°C and stirred at room temperature for 64 hours. Sodium bicarbonate is added at 0°C . The mixture is stirred at room temperature for 0.5 h and filtered. The filtrate is concentrated and the residue is purified by silica gel column chromatography to obtain compound PD-B. If an acid chloride or anhydride is used, a suitable base such as triethylamine is added in place of the drying agent.

PD-B-8i connection.

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To a solution of PD-A-8b (100 mg, 0.19 mmol) in DCM (1.0 mL) under argon at room temperature was added H,H-dimethylformamide dimethyl acetal (25 μL, 0.19 mmol). After 30 minutes, the reaction mixture is concentrated. The reaction was taken into DCM and concentrated. This process is repeated twice. The resulting residue was dissolved in THF (1.0 ml) and cooled to 0°C under argon. Triethylamine (79 μL, 0.57 mmol) and DMAP (5 mg, 0.04 mmol) were added to the solution. After 5 min, isobutyryl chloride (60 μL, 0.57 mmol) was added. After 10 minutes, the reaction mixture was warmed to room temperature and stirred for 3 hours. The mixture was cooled to 0°C, stopped with a 5% TFA solution in water, and then stirred at room temperature for 4 hours. The resulting mixture was extracted with ethyl acetate (3x). The combined organic layers were dried over sodium sulfate, filtered and concentrated. The residue was purified by RP HPLC (acetonitrile/water) to give PD-B-8i (71 mg, 61%). ^1H NMR (400 MHz, CDCl ₃ ): δ 8.17 (two s, 1H), 7.66 (two s, 1H), 7.34-7.14 (m, 5H), 5.69 (dd, 1H), 5.56-5.43 (m, 1H), 4.55-4.01 (m, 5H), 3.79-3.69 (m, 1H), 2.70-2.64 (m, 1H), 1.37-1.17 (m, 15H).

³¹ P NMR (161 MHz, CDCl ₃ ): δ 2.99 (s), 2.88 (s).

LC/MS=609 (M+H+).

Retention time: 2.21 min.

LC: Thermo Electron Surveyor HPLC.

MS: Finnigan LCQ Advantage MAX Mass Spectrometer.

Column: Phenomenex Polar RP 30 mm x 4.6 mm.

Solvents: acetonitrile with 0.1% formic acid, water with 0.1% formic acid.

Gradient: 0-0.1 min. 5% ACN, 0.1-1.95 min. 5-100% ACN, 1.95-3.5 min. 100% ACN, 3.5-3.55 min. 100-5% ACN, 3.55-4 min. 5% ACN.

The following example PD-B compounds were prepared following general procedures.

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General procedure for preparing a nucleoside prodrug (method E).

Non-limiting examples of 3',5'-cyclic monophosphoramidate prodrugs containing the present invention can be prepared according to General Scheme 3.

Scheme 3

Scheme 3 illustrates chemical processes that may be useful for preparing the PD-C compound. Accordingly, PD-A1 is converted to PD-C in the presence of a base when Ar is replaced by a leaving group electron such as a p-nitro or p-chloro group (European Journal of Medicinal Chemistry, 2009, 44, 3769). In addition, compound 40 is converted into compound 41 according to Bioorganic and Medicinal Chemistry Letters, 2007, 17, 2452, which is then, together with the amino acid ester salt

- 80 046452 forms PD-C.

Compound PD-C-8q.

A solution of PD-A-8p in DMSO is treated at room temperature with potassium tert-butoxide (~1 eq.) and the resulting mixture is stirred for about 10 minutes for about 2 hours. The mixture is then cooled to 0°C and neutralized with 1N HCl to ~pH 6. The mixture is purified by HPLC to give compound PD-C-8q.

In addition, the following exemplary PD-C compounds were prepared following general procedures.

1) ROS1z, Ri(OMe)z

KOSHCHVODN. ,ACN

Connection 8

Connection 41-1

Compound 8 is dissolved in PO(OMe) ₃ (0.1-0.5 M solution) and cooled to 0°C in an argon atmosphere. To do this, while stirring, POCl ₃ (1.0-5.0 eq.) was added dropwise to the solution, and the reaction mixture

Connection PD-C-8r

- 81 046452 was heated to room temperature for 2-16 hours. The resulting solution was added dropwise to a rapidly stirred solution of acetonitrile and 0.05-0.5 M aqueous solution of KOH. When the addition is complete, the solvents are removed under reduced pressure. The resulting residue was dissolved in water and purified by HPLC to give compound 41-1.

A solution of Compound 41-1 in DCM and PO(OMe)3 is prepared and cooled to 0°C. To this solution was added oxalyl chloride (1.0-5.0 eq.) followed by a catalytic amount of DMF. The mixture was stirred for about 10 minutes to about 1 hour. When fully activated, a large volume of 2-propanol was added to the reaction mixture and stirred and warmed to room temperature. Solvents were removed under reduced pressure and the resulting crude product was purified by preparative HPLC to give compound PD-D-8u.

Connection PD-E-8v.

Connection 41-1 Connection PD-E-8V

Compound PD-E-8v is prepared from compound 41-1 with a base similar to compound PD-D8u by replacing 2-aminopropane with 2-propanol.

Connection PD-F-8w.

Connection PD-F-8W

Compound PD-F-8w is prepared from the similar compound 20, replacing compound 8 with compound 18.

Connection PD-G-8x.

Connection PD-G-8X

About 90 mM of compound 8 in THF is cooled to -78°C, and about 2.2 to about 5 equivalents of tert-butylmagnesium chloride (about 1M in THF) are added. The mixture is heated to 0°C for 30 minutes and cooled again to -78°C. A solution of ^)-2-{[chloro(1-phenoxy)phosphoryl]amino ethyl isobutyrate (WO2008085508) (1M in THF, about 2 equivalents) was added dropwise. Cooling is stopped and the reaction mixture is stirred for about one to 24 hours. The reaction is quenched with water and the mixture is extracted with ethyl acetate. The extracts are dried and evaporated and the residue is purified by chromatography to give Compound PD-G-8x.

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Connection 6.

Compound 4 (about 0.04 mmol) and anhydrous methanol (about 5 ml) were treated with acetic acid (about 5 ml) and the reaction mixture was stirred overnight at room temperature. A saturated solution of NaHCO3 is added to neutralize the reaction mixture and the crude material is purified using an HPLC system (acetonitrile- _H2O ) to give 6.

Connection 7.

Compound 3b (about 0.39 mmol) and anhydrous dichloromethane (about 10 ml) were added to a dry argon-flushed round bottom flask (50 mL). The flask is placed in a dry ice/acetone bath (-78°C) and the solution is stirred for approximately 10 minutes. BF ₃ -Et ₂ O (about 0.10 ml) is added dropwise and the reaction mixture is stirred for 10 minutes. AlMe ₃ (about 1.16 mmol, 2.0 M in toluene) is then added. After a few minutes, the dry ice/acetone is removed and the reaction mixture is stirred at room temperature to approximately 45°C for approximately 4 hours to 4 days. A solution of pyridine (about 2 ml) in methanol (about 10 ml) is added and the solvent is removed under reduced pressure. The crude product is purified by chromatography and treated with ammonium hydroxide in methanol for 16 hours at room temperature. The mixture is concentrated and the residue is purified by HPLC to give 7.

Connection 8.

To a suspension of 7-bromimidazo[1,2-L| [1,2,4]triazin-4-amine (according to ACS Medicinal Chemistry Letters, 2010, 1, 286; 375 mg, 1.75 mmol) in THF (4.0 ml) was added under argon. 2-bis-[(chlorodimethyl)silanyl]ethane (452 mg, 2.10 mmol). After 60 min, the reaction mixture was cooled to -78°C and BuLi (1.6 M in THF, 3.8 mL, 6.10 mmol) was added. After 10 min at -78°C, a solution of 1a (prepared according to WO 200631725, 782 mg, 2.10 mmol) in THF (1.0 ml) was added dropwise. The resulting mixture was stirred at -78°C for 1 hour. A saturated aqueous solution of ammonium chloride was added and heated to 0°C. Add water until all solids are soluble. The mixture is extracted with ethyl acetate. The organic extract is dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (ethyl acetate/hexane) to give 8b (606 mg, 59%) as a yellow solid.

LC/MS=508 (M+H+).

Retention time: 2.17-2.26 min.

LC: Thermo Electron Surveyor HPLC.

MS: Finnigan LCQ Advantage MAX Mass Spectrometer.

Column: Phenomenex Polar RP 30 mm x 4.6 mm.

Solvents: acetonitrile with 0.1% formic acid, water with 0.1% formic acid.

Gradient: 0-0.1 min. 5% ACN, 0.1-1.95 min. 5-100% ACN, 1.95-3.5 min. 100% ACN, 3.5-3.55 min. 100-5% ACN, 3.55-4 min. 5% ACN.

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To a solution of 8b (510 mg, 1.39 mmol) in dichloroethane (10.0 mL) at 0°C under argon, add silane triethylamine (1.77 mL, 11.09 mmol) followed by BF ₃ -Et ₂ O (1.41 ml, 11.09 mmol). The reaction mixture is stirred at 55°C for 16 hours. The reaction mixture is cooled to 0°C and stopped with saturated NaHCO ₃ (aq). The reaction products are extracted with DCM and then EtOAc. The combined organic phase is dried over sodium sulfate, filtered and concentrated. The residue was purified by silica gel chromatography (ethyl acetate/hexane) to give 8c (453 mg, 64%). ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.10-7.94 (m, 5H), 7.6-7.33 (m, 7H), 5.91 (dd, 1H), 5.78 (d, J=24.6 Hz, 1H), 4.87 (dd, 1H), 4.70 (m, 1H), 4.58 (dd, 1H), 1.31 (d, J=22.4 Hz, 3H).

LC/MS=491 (M ⁺ ).

Retention time: 2.36 min.

LC: Thermo Electron Surveyor HPLC.

MS: Finnigan LCQ Advantage MAX Mass Spectrometer.

Column: Phenomenex Polar RP 30 mm x 4.6 mm.

Solvents: acetonitrile with 0.1% formic acid, water with 0.1% formic acid.

Gradient: 0-0.1 min. 5% ACN, 0.1-1.95 min. 5-100% ACN, 1.95-3.5 min. 100% ACN, 3.5-3.55 min.

100-5% ACN, 3.55-4 min. 5% ACN.

c Connection 8

To a solution of 8c (500 mg, 01.02 mmol) in THF (5.0 ml) add lithium hydroxide (122 mg, 5.09 mmol) as a solution in H ₂ O (5.0 ml) and stir at room temperature within 1 hour. The reaction mixture is cooled to 0°C and neutralized with 1N HCl in water (5.1 ml). The mixture was concentrated and the residue was purified by RP HPLC (water/acetonitrile) to give compound 8 (185 mg, 64%). 1H NMR (400 MHz,

CD ₃ OD): δ 7.97 (s, 1H), 7.63 (s, 1H), 5.54 (d, J=24.8 Hz, 1H), 4.03 (dd, 1H), 3.88 (m, 1H), 3.71 (dd, 1H), 1.80 (d, J=22.1 Hz, 3H).

LC/MS=284 (M+H ⁺ ).

Retention time: 1.06 min.

LC: Thermo Electron Surveyor HPLC.

MS: Finnigan LCQ Advantage MAX Mass Spectrometer.

Column: Phenomenex Polar RP 30 mm x 4.6 mm.

Solvents: acetonitrile with 0.1% formic acid, water with 0.1% formic acid.

Gradient: 0-0.1 min. 5% ACN, 0.1-1.95 min. 5-100% ACN, 1.95-3.5 min. 100% ACN, 3.5-3.55 min. 100-5% ACN, 3.55-4 min. 5% ACN.

Alternative procedure for connection 8.

Compound 1e (crude obtained from the previous reaction step) is dissolved in ethanol. Excess sodium borohydride is added in portions until the reaction is almost complete. The mixture is neutralized with acetic acid. The mixture was concentrated and the solid was purified by silica gel column chromatography (0-10% methanol/dichloromethane) to give compound 27 (210 mg, 50% in two steps).

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Additional alternative procedure for connection 8.

NO F NO F

1d 8

Raney nickel (ca. 500 mg) was neutralized by washing with H2O, and added to a solution of 1d (ca. 100 mg) in ethanol (ca. 10 ml). The mixture was then heated to 80°C until the reaction was complete. The catalyst is removed by filtration and the solution is concentrated in vacuo. The mixture was concentrated and the residue was purified by HPLC to give 8.

Connection 9.

NH ₂

II?

HO—R—O—w Q VN _S he \ / ' ^{CN N}

BUT F

To the flask containing compound 3 (120 mg, 0.39 mmol, 1 eq.) add PO(OMe) ₃ (1.5 ml, 0.25 M) and cool to 0°C before adding POCl ₃ (125 µl, 1 .37 mmol, 3.5 eq.). The reaction mixture is stirred for 5 hours until the reaction stops with water. It was directly purified by HPLC to isolate the monophosphate of compound 9.

LC MS m/z 387.95 [M+H+].

Connection 10.

o nh ₂ \ο^ο^ Π / o—r—O—f ₀ d

O\V-'CN ^N \d° N)—O'L but F / O

To a flask containing compound 9 (30 mg, 0.078 mmol, 1 eq.) add NMP (0.8 ml, 0.1 M) followed by TEA (43 μl, 0.31 mmol, 4 eq.), tetrabutylammonium bromide (25 mg, 0.078 mmol, 1 eq.) before adding chloromethyl isopropyl carbonate (60 μl, 0.38 mmol, 5 eq.). The reaction mixture is heated to 50°C and stirred overnight. This was purified directly by high performance liquid chromatography to give compound 10.

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.98 (s, 1H), 7.01 (d, J=4.7 Hz, 1H), 6.72 (d, J=4.7 Hz, 1H), 6.04 (bs, 2H), 5.74 -5.61 (m, 4H), 4.91 (ddt, J=12.6, 9.4, 6.3 Hz, 2H), 4.64-4.28 (m, 4H), 1.37-1.19 (m, 15H).

³¹ P NMR (162 MHz, CDCl ₃ ) δ -4.06.

¹⁹ F NMR (376 MHz, CDCl ₃ ) δ -76.58, -151.95 TFA salt.

LC MS m/z 620.03 [M+H+].

Connection 11.

A solution of compound B-2 in DMSO is treated with about 3 mol equivalents of potassium tert-butoxide for 15 minutes to 24 hours. The reaction is stopped with 1N HCl and compound 11 is isolated by reverse phase HPLC.

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Connection 12.

About Bz

o f

Bz

NH ₃

NHO Bz

F

Bz

1b

12a

Nickel Raney

HO

HO F nh ₂

1.TMSCN

TMSOTf

2. NH ₃

BzO

BzO F

12b nh ₂

Compound 1b (about 1 mmol) was placed in a steel bomb reactor. The reactor is charged with liquid ammonia (about 30 ml) and the mixture is stirred at 0°C to about 50°C for about 16 hours. The ammonia is evaporated and the residue is purified to give 12a. A solution of 12a (about 100 mg) in ethanol (about 10 ml) is treated with Raney nickel (about 500 mg), which is neutralized by washing with H ₂ O. The mixture is then heated to about 35 to about 80° C. until the reaction is complete. The catalyst is removed by filtration and the solution is concentrated in vacuo. The mixture was concentrated and the residue was purified by HPLC to give 12b. To a solution of compound 12b (about 50 mg) and TMSCN (about 0.5 mmol) in acetonitrile (about 2.0 ml) at about 0°C add TMSOTf (about 0.5 mmol). The reaction mixture is stirred at room temperature for 1 hour, then at 65°C for 3 days. The reaction is stopped with saturated NaHCO ₃ at room temperature and the mixture is diluted with CH ₃ CO2Et. The organic phase is separated by washing with brine, dried over Na ₂ SO ₄ , filtered and concentrated. The residue is purified by RP-HPLC, then dissolved in methanol (about 1 ml). Ammonium hydroxide (28% in water, about 0.8 ml) was added and the mixture was stirred at room temperature for 16 hours. The mixture was concentrated and the residue was purified by RP HPLC to give 12.

Connection 13.

Compound 13 was prepared in the same manner as Compound 9 using Compound 12 as the starting material.

Connection 14.

Compound 14 is prepared by treating compound 13 with about one to about five equivalents of DCC in pyridine and heating the reaction mixture to boiling for about one to about 24 hours. Compound 14 is isolated by conventional ion exchange and reverse phase HPLC.

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Connection 15.

O—x ' < °-P-0 r

ABOUT

A solution of about 0.4 mmol of compound 14 in about 10 ml of DMF is treated with about 0.8 mmol of DIPEA and about 0.8 mmol of chloromethyl isopropyl carbonate (WO2007/027248). The reaction mixture is heated to about 25 to about 80°C for about 15 minutes to about 24 hours. The solvent is removed in vacuo and the residue is purified by HPLC to give compound 15.

Connection 16.

NHDMTr n ISK Vn J ^N but' ¥

Compound 3 (about 0.22 mmol) was dissolved in anhydrous pyridine (about 2 ml) and chlorotrimethylsilane (about 0.17 ml). The mixture is stirred at a temperature of about 0°C to 25°C for about one to about 24 hours. Additionally, chlorotrimethylsilane (about 0.1 ml) is added and the reaction mixture is stirred for about one to about 24 hours. dimethoxytrityl chloride (about 0.66 mmol) and DMAP (about 0.11 to about 0.22 mmol) are added sequentially. The mixture is stirred for one to about 24 hours. A solution of TBAF (1.0 M, about 0.22 mL) in THF is added and the reaction mixture is stirred for about one to about 24 hours. The mixture is partitioned between ethyl acetate and water. The ethyl acetate layer is dried and concentrated. The residue is purified by chromatography to give compound 16.

Connection 17.

A mixture of about 1.25 mmol of compound 16 and

about 1.9 mmol of triethylammonium 2-(2,2-dimethyl-3(trityloxy)propanoylthio)ethylphosphinate (WO2008082601) was dissolved in anhydrous pyridine (about 19 ml). Pivaloyl chloride (about 2.5 mmol) is added dropwise at about -30 to about 0°C and the solution is stirred for about 30 minutes to about 24 hours. The reaction mixture is diluted with methylene chloride and neutralized with aqueous ammonium chloride (about 0.5 M) . The methylene chloride phase is evaporated and the residue is dried and purified by chromatography to give compound 17.

Connection 18.

To a solution of about 0.49 mmol of compound 17 in anhydrous carbon tetrachloride (about 5 ml) benzylamine (about 2.45 mmol) was added dropwise. The reaction mixture is stirred for one to about 24 hours. The solvent is evaporated and the residue is purified by chromatography to give compound 18.

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Connection 20.

A solution of about 2 mmol of compound 18 in methylene chloride (about 10 ml) is treated with an aqueous solution of trifluoroacetic acid (90%, about 10 ml). The reaction mixture is stirred at a temperature of about 25 to about 60°C for one to about 24 hours. The reaction mixture is diluted with ethanol, the volatiles are evaporated and the residue is purified by chromatography to give compound 20.

Connection 21.

Approximately 90 mM of compound 2 was cooled in THF for several minutes at approximately -78°C and approximately 2.2 to approximately 5 equivalents of t-butylmagnesium chloride (about 1M in THF) was added. The mixture was heated to approximately 0°C for approximately 30 minutes and cooled again to approximately -78°C.

A solution of DO)-2-{[chloro(1-phenoxy)phosphoryl]amino}propyl pivaloate (WO2008085508) (1M in THF, about 2 equivalents) is added dropwise. Cooling is stopped and the reaction mixture is stirred for about one to about 24 hours. The reaction is quenched with water and the mixture is extracted with ethyl acetate. The extracts are dried and evaporated and the residue is purified by chromatography to give compound 21.

Connection 22.

Compound 22a was prepared in a manner similar to that for C-1a. 1H NMR (400 MHz, CDCl ₃ ) δ 8.11 (d, J=9.0 Hz, 2H), 8.02 (s, 1H), 7.48 (t, J=7.5 Hz, 2H), 7.42-7.25 (m, 4H), 7.21 (dt, J=14.9, 5.5 Hz, 2H), 7.08 (t, J=7.3 Hz, 2H), 5.17-5.03 (m, 2H), 4.99 (dd, J=16.5, 9.7 Hz, 2H), 3.44 (s, 1H), 3.35-3.21 (m, 2H), 3.19 (d, J=9.2 Hz, 1H), 3.00-2.80 (m, 2H). ³¹ P NMR (162 MHz, CDCl ₃ ) δ 4.27. LC MS m/z 452.09 [M+H+].

Compound 22b was prepared in a manner similar to the preparation of C-1 using compounds 3 and 22a.

1H NMR (400 MHz, CD3OD) δ 7.76 (d, J=6.3 Hz, 1H), 7.38 (t, J=8.2 Hz, 1H), 7.27-7.12 (m, 4H), 7.066.81 (m, 3H) , 6.74 (dd, J=4.6, 3.5 Hz, 1H), 4.95-4.79 (m, 1H), 4.35-3.90 (m, 4H), 3.23 (dt, J=3.2, 1.6 Hz, 3H), 3.18-3.05 (m, 2H), 2.82 (dt, J=14.7, 7.3 Hz, 2H), 1.15 (d, J=22.4 Hz, 3H). ³¹ P NMR (162 MHz, CD3OD) δ 10.76,

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10.71. LC MS m/z 620.05 [M+H+].

To a flask containing 22b (50 mg, 0.08 mmol, 1 eq.) add ethanol (4 ml), then Pd(OH) ₂ (56 mg, 0.08 mmol, 1 eq.) and ammonium formate (42 mg , 0.64 mmol, 8 eq.). The reaction mixture is heated to 80°C for one hour. The solid is filtered off and the material is purified by HPLC.

¹ H NMR (400 MHz, DMSO-d6) δ 10.72 (s, 1H), 7.91 (s, 1H), 7.95-7.89 (bs, 2H), 7.41 (d, J=7.7 Hz, 1H), 7.26 (d , J=8.1 Hz, 1H), 7.19-6.66 (m, 3H), 4.20-3.75 (m, 3H), 2.99 (dd, J=16.5, 9.6 Hz, 2H), 2.89-2.70 (m, 2H), 2.48-2.58 (m, 8H), 1.10 (d, J=22.3 Hz, 3H). ³¹ P NMR (162 MHz, DMSO-d,) δ 7.49. ¹⁹ F NMR (376 MHz, DMSO-d6) δ -154.89. LC MS m/z 530.21 [M+H+].

Connection 23.

Compound 3 (250 mg, 0.82 mmol) was dissolved in PO(OMe) ₃ (5 ml, 0.16 M) and cooled to 0°C under argon. For this stirring, POCl ₃ (0.32 ml, 4.1 mmol) is added slowly dropwise and the reaction mixture is warmed to room temperature over 16 hours. The resulting solution is added dropwise with rapid stirring to the acetonitrile solution (400 ml) and 0.08 M aqueous KOH (300 ml). When the addition was completed, the progress of the reaction was checked by LCMS. When the reaction has been completed, the solvent is removed under reduced pressure. The resulting solid was dissolved in water and purified by HPLC to obtain 140 mg of compound 23 (yield: 47%).

¹ H-NMR (400 MHz; CD3OD): δ 8.15 (s, 1H), 7.40 (d, 1H; J=4.8 Hz), 7.09 (d, 1H; J=4.8 Hz), 4.64 (dd, 1H; J =24 Hz, 7.2 Hz), 4.50-4.36 (m, 3H), 1.32 (d, 3H; J=22 Hz).

^F-NMR (376 MHz; CD3OD): δ -153.11.

³¹ P-NMR (162 MHz; CD ₃ Od): δ -2.20.

MS [M+H+]=370.2.

Connection 24.

A solution of compound 23 (7 mg, 0.02 mmol) in DCM (2 ml) and PO(OMe) ₃ (1 ml) was prepared and cooled to 0°C. To this solution add oxalyl-Cl (10 µl) followed by DMF (2 µl). The mixture is stirred for 1 minute, an aliquot is removed and cooled in methanol and then checked by LCMS for activation. Add successive amounts of oxalyl-Cl (10 µl) and DMF A (2 µl) until complete activation. At this point, a large volume of 2-propanol (5 ml) is added to the reaction mixture and stirred and warmed to room temperature. After completion of the reaction, the solvent was removed under reduced pressure and the resulting crude product was purified by preparative HPLC to obtain 5.5 mg of compound 24 (70% yield). 1H-NMR (400 MHz; DMSO-d ₆ ): δ 8.26 (br, 1H), 8.15 (br, 1H), 7.97 (s, 1H), 7.00 (d, 1H; J=4.4 Hz), 6.88 (d , 1H; J=4.4 Hz), 4.59-4.51 (m, 2H), 4.37-4.25 (m, 2h), 1.23 (d, 3H; J=22.8 Hz). ^F-NMR (376 MHz; CD3OD): δ -151.72. ³¹ P-NMR (162 MHz; CD3OD): δ -5.69. MS [M+H+]=412.0.

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Connection 25.

Compound 25 is prepared from compound 23, which is essentially similar to compound 24, replacing alanine heptyl ester with 2-propanol (5.3% yield). ¹ H-NMR (400 MHz; CD ₃ OD): δ 7.91 (s, 1H), 6.98 (d, 1H; J=4.8 Hz), 6.92 (d, 1H; J=4.8 Hz), 5.29 (dd, 1H ; J=24.4 Hz, 8.8 Hz), 4.66-4.60 (m, 2H), 4.48-4.40 (m, 1H), 4.15-4.11(m, 3H), 3.92 (dd, 1H; J=9.6 Hz, 7.2 Hz ), 1.67-1.64 (m, 3H), 1.40-1.27 (m, 15H), 0.91-0.87 (m, 6H). ¹⁹ P-NMR (376 MHz; CD3OD): δ -151.46. ³¹ P-NMR (162 MHz; CD3OD): δ 7.36. MS [M+H+]=539.4.

Connection 26.

Connection 26

Compound 26 is prepared from compound 22 in a manner substantially similar to the preparation of compound 10.

Antiviral activity.

Another aspect of the invention provides methods for inhibiting viral infections, comprising the step of treating a sample or item suspected of needing such inhibition with a composition of the invention.

In the context of the present invention, samples suspected of containing viruses include natural or artificial materials such as living organisms; tissue or cell cultures; biological samples, such as biological material samples (blood, serum, urine, cerebrospinal fluid, tears, sputum, saliva, tissue samples, etc.); laboratory samples; samples of food, water or air; bioproducts of samples, such as an extract of cells, in particular cells synthesizing the recombinant desired glycoprotein, and the like. Typically, the sample will be expected to contain the organism that causes the viral infection, often a pathogenic organism such as a tumor virus. Samples may be maintained in any medium, including water and organic solvent/water mixtures. Samples include living organisms such as humans and artificial materials such as cell cultures.

If desired, the antiviral activity of a compound of the invention after application of the composition can be observed by any method, including direct and indirect methods of detecting such activity. Quantitative, qualitative and semi-quantitative methods for determining such activity are all considered. Typically, one of the screening methods described above is used, however, any other methods, such as observing the physiological properties of a living organism, are also applicable.

The antiviral activity of a compound of the invention can be measured using standard screening protocols that are known. For example, the antiviral activity of a compound can be measured using the following general protocols.

Cellular Flavivirus immunoassay.

BHK21 or A549 cells are trypsinized, counted and diluted to 2x105 cells/ml in Hams F-12 medium (A549 cells) or RPMI-1640 medium (BHK21 cells) supplemented with 2% fetal bovine serum (FBS) and 1% penicillin/streptomycin . 2 x 104 cells per well are plated into wells of a 96-well tissue culture plate and left at 37°C, 5% CO ₂ overnight. The next day, cells infected with viruses at a multiplicity of infection (MOI) of 0.3 in the presence of varying concentrations of test compounds were incubated for 1 hour at 37°C and 5% CO ₂ and left for an additional 48 hours. Cells were washed once with PBS and Fix with cold methanol for 10 minutes. After washing twice with PBS, fixed cells are blocked with PBS containing 1% FBS and 0.05% Tween-20 for 1 h at room temperature. Primary antibody solution (4G2) is then added at a concentration of 1:20 to 1:100 in PBS containing 1% FBS and 0.05% Tween-20 and incubated for 3 hours. The cells are then washed three times with PBS, followed by one hour of incubation with horseradish peroxidase (HRP)-conjugated anti-mouse IgG antibodies (Sigma, diluted 1:2000). By

- 90 046452 After washing three times with PBS, 50 µl of 3,3',5,5'-tetramethylbenzidine (TMB) substrate solution (Sigma) is added to each well for two minutes. The reaction is stopped by adding 0.5 M sulfuric acid. The plates are read at 450 nm absorbance to quantify viral load. After measurement, cells are washed three times with PBS followed by incubation with propidium iodide for 5 min. The plates are read in a Tecan Satire™ photometer (excitation 537 nm, emission 617 nm) for cell quantification. Dose-response curves are plotted based on the average optical density versus the tenth logarithm of the concentration of the test compounds. EC ₅₀ is calculated by nonlinear regression analysis. A positive control such as N-nonyldioxynoirimicin can be used.

Cellular analysis of the cytopathic effect of Flavivirus.

To test for West Nile virus or Japanese encephalitis virus, BHK21 cells are trypsinized and diluted to a concentration of 4 x 10 ⁵ cells/ml in RPMI-1640 medium with 2% FBS and 1% penicillin/streptomycin. For testing against dengue virus, Huh7 cells are trypsinized and diluted to a concentration of 4 x 10 ⁵ cells/ml in DMEM supplemented with 5% FBS and 1% penicillin/streptomycin. 50 μl of cell suspension (2 x 10 ⁴ cells) is dispensed per well into 96-well polymer-based plates, optically accessible from the bottom of the PIT (Nunc). Cells are grown overnight in culture medium at 37°C, 5% CO ₂ and then infected with West Nile virus (eg, strain B956) or Japanese encephalitis virus (eg, Nakayama strain) at an MOI of 0.3, or with virus dengue (for example, DEN-2 NGC strain) at MOI=1, are studied in the presence of various concentrations of the test compounds. The plate containing the virus and compounds is further incubated at 37°C, 5% CO ₂ for 72 hours. At the end of the incubation, 100 μl of CellTiter-Glo™ reagent is added to each well. The contents are mixed for 2 minutes on an orbital shaker to cause cell lysis. The plates are incubated at room temperature for 10 min to stabilize the luminescent signal. Luminescence is read from the recorded plates using a plate reader. A positive control such as N-nonyldioxynoirimicin can be used.

Antiviral activity in a mouse model of dengue infection.

The compounds are being tested in vivo in a mouse model of dengue virus infection (Schul et al. J. Infectious Dis. 2007; 195:665-74). Six to ten week old AG129 mice (B&K Universal Ltd, Hll, UK) were housed in individual ventilated cages. Mice are injected intraperitoneally with 0.4 ml of TSV01 dengue virus 2 suspension. Blood samples were collected through a retro-orbital puncture under isoflurane anesthesia. Blood samples were collected into tubes containing sodium citrate to a final concentration of 0.4% and immediately centrifuged for 3 min at 6000 g to obtain plasma. Plasma (20 μl) was diluted in 780 μl RPMI-1640 and frozen in liquid nitrogen until subsequent epidemic analysis. The remaining plasma was reserved for determination of cytokine and NS1 protein levels. The mice developed dengue fever with increasing viremia over several days, reaching a maximum on the 3rd day after infection.

To test antiviral activity, compounds of the invention were dissolved in a carrier liquid, for example, 10% ethanol, 30% PEG-300 and 60% D5W (5% dextrose in water, or 6 N HCl (1.5 eq): 1 N NaOH (pH up to 3.5): 100 mM citrate buffer pH 3.5 (0.9% v/v: 2.5% v/v: 96.6% v/v). Six 6-10 week old AG129 mice were divided into six groups of six mice each. All mice were infected with dengue virus as described above (day 0). Group 1 was gavaged with 200 ml/mouse with 0.2 mg/kg of compound. invention twice daily (once early in the morning and once in the afternoon) for three days, starting on day 0 (first dose immediately before dengue infection Groups 2, 3 and 4 are dosed in the same way, with 1 mg). /kg, 5 mg/kg and 25 mg/kg of the compound, respectively. A positive control can be used, for example, (2R, 3R, 4R, 5R)-2-(2-amino-6-hydroxy-purin-9-yl). )-5-hydroxymethyl-3-methyl-tetrahydro-furan-3,4-diol, dosed by gavage 200 µl/mouse in the same way as in previous groups. The other group is considered only with a carrier liquid.

On day 3 postinfection, approximately 100 μl of blood (anticoagulated with sodium citrate) was collected from mice by retroorbital puncture under isoflurane anesthesia. Plasma was obtained from each blood sample by centrifugation and frozen in liquid nitrogen for subsequent epidemic analysis. Collected plasma samples were analyzed by epidemic assay as described by Schul et al. Cytokines were also analyzed as described by Schul. NS1 protein levels were analyzed using the Platelia™ kit (BioRad Laboratories). The antiviral effect is indicated by a decrease in the level of cytokines and/or NS1 protein.

Typically, a reduction in viremia of about 5-100-fold, more typically 10-60-fold, most commonly 20-30-fold, is obtained at 5-50 mg/kg dose volume of the compounds of the invention.

Determination of HCV IC ₅₀ .

Assay protocol: either wild type or S282T (Migliaccio, et al., J. Biol. Chem. 2003, 49164-49170; Klumpp, et al., J. Biol. Chem. 2006, 3793-3799) mutant polymerase enzyme was used in this

- 91 046452 analysis. NS5b polymerase assay (40 μl) was assembled by adding 28 μl of polymerase mixture (final concentration: 50 mM Tris-HCl at pH 7.5, 10 mM KCl, 5 mM MgCl ₂ , 1 mM DTT, 10 mM EDTA, 4 ng /μl RNA template, and 75 nM HCV A 21 NS5b polymerase) for plate assay, then 4 μl of dissolved compound was added. The polymerase and compound were preincubated at 35°C for 10 min before adding 8 μl of a nucleotide substrate mixture (33P-α-labeled nucleotide at a competitive K _m value and 0.5 mM of the remaining three nucleotides). The assay plates were sealed and incubated at 35°C for 90 min. The reaction mixtures were then filtered through 96 and DEAE-81 filter plates using vacuum. The filter plates were then washed under vacuum with several volumes of 0.125 M NaHPO ₄ , water and ethanol to remove unincorporated labels. The plates were then analyzed on TopCount to assess the level of synthesis products compared to background controls. IC ₅₀ volume was determined using the established Prism program.

Preferably, the compounds described herein inhibit NS5b polymerase with an IC ₅₀ below 1000 μM, more preferably below 100 μM, and most preferably below 10 μM. For example, compound TP-1 has an IC ₅₀ of 0.15 µM against both wild-type HCV polymerase and the S282T mutant enzyme. Table II below shows the activity of TP-1 and TP-2 against both the wild type and the S282T mutant enzyme compared to the activity obtained with 2'-methylguanidine triphosphate and (2R,3R,4R,5R)-2-(4-aminopyrrolo) triphosphate [1,2-f][1,2,4]triazin-7-yl)-3,4-dihydroxy-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-carbonitrile. This suggests that replacing the 2'OH pyrrolo[1,2£][1,2,4]triazin-7-yl nucleosides with 2'F unexpectedly confers activity against resistant S282T HCV mutant strains of the virus.

Table II

Triphosphate WT IC ₅ o(mkM) S282T IC ₅ o(mkM) Note OOO II II II but-r-o-r-o-r-o- HE HE HE Λ,Ο N ^- but it is 2'-C-MeGTP 0 1/ NH ⁿ⁴ nh ₂ 0.1 20 from J. Bio. Chem., 2003,278,49164 (200 fold shift) LLC II II II N-R-O-R-O-R-O he he he O LJnh \ ^N nh ₂ H0 > TP-1 0.15 0.15 (1x shift) OOO II II II N-R-O-R-O-R-O he he he CM Garden ο ^ζ J 0.525 111 WO/2009/132135 (242 fold shift) LLC II II II N-R-O-R-O-R-O- he he he LLC II II II N-R-O-R-O-R-O- he he he but he nh ₂ to V Ύ, \ //CN HO F TP-2 nh ₂ ΛΑ z ^N ' V / N- ⁷ 0.24 0.034 1.60 (7x shift) HO F TP-3 LLC II II II N-R-O-R-O-R-O he he he HO F TP-8a nh ₂ hl · J -F> 0.30 1.6 (5.3x shift)

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Determination of HCV EC50.

Replicon cells were seeded in 96-well plates at a density of 8x10 ³ cells per well in 100 μl of culture medium, with the exception of Geneticin. The compound was serially diluted in 100% DMSO and then added to the cells at a 1:200 dilution to achieve a final concentration of 0.5% DMSO and a total volume of 200 μl. The plates were incubated at 37°C for 3 days, after which the culture medium was removed and the cells were lysed in lysis buffer provided by the Promega Luciferase Assay System. Following the manufacturer's instructions, 100 μl of luciferase substrate was added to the lysed cells and luciferase activity was measured in a TopCount luminometer. Preferably, the compounds described herein have an EC ₅₀ below 1000 μM, more preferably below 100 μM, and most preferably below 10 μM. The activities of the presented compounds of formula I are given in table. III below.

The cytotoxicity of a compound of the invention can be determined using the following general protocols.

Metabolism study.

Applicants have noted that monophosphate prodrugs of nucleoside analogues with a nitrogen at the X ¹ position may have increased activity compared to their partners with a carbon at the X ¹ position. This difference in activity correlates with the amount of active triphosphate analog compounds in the cells. This can be quantified by metabolic studies, which quantify the intracellular concentrations of triphosphate analogues. Increased intracellular concentrations of the triphosphate metabolite correlate with a prodrug with increased activity.

For example, comparison of the prodrug of compound B-7 with the prodrug of compound PD-A-8b shows increased activity when the X ¹ position is nitrogen. This can be seen in table. III, where the HCV EC ₅₀ for the compound where the X ¹ nitrogen position (compound PD-A-8b) is 0.68 µM compared to 63-73 µM for compound B-7. Activation of the prodrug analogue PD-A-8b (its triphosphate analogue TP-8a) showed that it is more than two orders of magnitude more effective than that observed for its prodrug counterpart, where the X ¹ position is carbon, B-7 (in its triphosphate analogue TP-3), as shown in table. IV.

Experimental data.

Huh-luc/neo replicons cells containing HCV genotype 1b subgenomic replicons were maintained in Dulbecco's modified Eagle's medium containing Glutamax supplemented with 10% heat-inactivated fetal bovine serum, penicillin-streptomycin and G418 disul.

- 93 046452 veil of saline solution. Cells were transferred to a twelve-well tissue culture plate, tryponized, and grown to confluence (0.88 x 106 cells/well). Cells were treated for 24 h with 10 μM nucleosides, or 10 μM prodrug. After 24 h, cells were washed 2 times with 2.0 ml of ice-cold 0.9% saline sodium chloride. Cells were then damaged with 0.5 ml of 70% methanol (MeOH) and frozen overnight to facilitate extraction of nucleotide metabolites. The extracted cell material in 70% MeOH was transferred into tubes and dried. After drying, samples were resuspended in 1 mM ammonium phosphate, pH 8.5, containing internal standard (100 nM ClATP). Intracellular levels of nucleoside triphosphates were quantified from authentic standard curves using liquid chromatography coupled to tandem mass spectrometry.

Results.

Table IV

Intracellular concentrations of triphosphate analogues formed in Huh-luc/neo replicon cells _____________ after 24 h of incubation with 10 μM PD-A-8b and B-7_____________

Prodrug Triphosphate Intracellular concentration of triphosphate analog (µM/million) AT 7 TR-3 <0.11 ^a PD-A-8b TR-8a 20.5 ^{b a} Intracellular concentrations were below the lower limit of quantitation. ^b The value is the average of 2 individual wells.

Cell culture cytotoxicity assay (CC50 determination).

The assay is based on the evaluation of the cytotoxic effect of the test compounds using a metabolic substrate. Analysis protocol for CC50 determination.

1. Maintain MT-2 cells in RPMI-1640 medium supplemented with 5% fetal bovine serum and antibiotics.

2. Dispense cells into a 96-well plate (20,000 cells in 100 µl media per well) and add different concentrations of test compound in triplicate (100 µl/well). Enable raw control.

3. Incubate cells for 5 days at 37°C.

4. Prepare XTT solution (6 ml for plate assay) in the dark at a concentration of 2 mg/ml in phosphate buffered saline pH 7.4. Heat the solution in a water bath at 55°C for 5 minutes. Add 50 µl N-methylphenazonium metasulfate (5 µg/ml) to 6 ml XTT solution.

5. Remove 100 µl of medium from each well of the assay plate and add 100 µl of XTT substrate solution per well. Incubate at 37°C for 45 to 60 minutes in a CO ₂ incubator.

6. Add 20 µl of 2% Triton X-100 per well to stop the metabolic conversion of XTT.

7. Read absorbance at 450 nm with background subtraction at 650 nm.

8. Plot the percentage of absorption compared to the untreated control and estimate the CC50 value as the drug concentration resulting in 50% inhibition of cell growth. Consider uptake to be directly proportional to cell growth.

All publications, patents and patent documents cited herein above are incorporated herein by reference, as if individually, by reference. The invention has been described with reference to various specific and preferred embodiments and methods. However, those skilled in the art will appreciate that many changes and modifications can be made while remaining within the spirit and scope of the invention.

Claims (3)

1. Compound of formula V or a pharmaceutically acceptable salt thereof; where R ¹ represents C1- _C8 alkyl; R ^a represents -C(=O)R11; R ⁷ represents Y represents O; W ¹ and W ² each independently represent a group of formula IVa R ^x formula IVa where each Y ¹ represents O; each Y ² represents O or NR; M2 represents 0; each R ^x represents a group of formula IVb M1s M1a formula IVb where each M1a, M1c and M1d represents 0; M12c represents 0 or 1; each R ^y independently represents H, -C(=Y1)OR, -OC(=Y1)R, C ₁ -C ₈ -alkyl, C ₆ -C2 ₀ -aryl or C ₇ -C ₂₀ -arylalkyl; where each C ₆ -C ₂₀ -aryl in R ^y is unsubstituted or substituted by 1-3 R ²⁰ groups; and each R is, independently, H, C1- _C8 alkyl or _C3 - _C20 carbocyclyl; wherein each C _1-8 alkyl in R is optionally substituted with one or more hydroxy; R ⁸ represents NH2; each R ¹¹ represents, independently, H or C1-C ₈ -alkyl; and each R ²⁰ is, independently, halogen. 2. Pharmaceutical composition for the treatment of infection caused by the Flaviviridae virus, containing - 95 046452 providing a therapeutically effective amount of a compound according to claim 1 and a pharmaceutically acceptable carrier. 3. The pharmaceutical composition according to claim 2, in which the compound is selected from the group consisting of Compound PD-B-8j nh ₂ PD-B-8k connection Connection PD-B-8I nh ₂ - 96 046452 Connection PD-B-8m Connection PD-B-8n Connection PD-B-80 nh ₂ and a pharmaceutically acceptable salt of any of these compounds.