ANTIVIRAL PHOSPHINATE COMPOUNDS

10

FIELD OF THE INVENTION

This application relates generally to compounds and pharmaceutical 15 compositions useful for the treatment of infection by, but not limited to, RNA viruses. In particular, these derivatives are intended for the treatment of infection by the Hepatitis C virus (HCV).

BACKGROUND OF THE INVENTION

20 Hepatitis C virus (HCV) is the major cause of non-A non-B hepatitis worldwide. Infection with HCV can progress to chronic liver disease (chronic hepatitis C), which can then progress to serious conditions such as liver cirrhosis,, hepatocellular carcinoma and terminal liver disease leading to death.

The current standard-of-care treatment for HCV infection is interf eron-α

25 (or its PEG-derivatized equivalent) in combination with ribavirin, a regimen that

produces sustained virologic response in only 40% of people infected with the HCV genotype 1. This regimen has significant side effects, leading an unacceptable number of patients to discontinue treatment (Hepatology, 2002, 2, 205). There is a clear need for novel therapies that are both more effective, and more tolerable to treat patients

10 Therefore, a need exists for the development of effective antiviral agents for treatment of HCV infection that overcomes the limitations of existing pharmaceutical therapies.

SUMMARY OF THE INVENTION

15 The present application is directed to compounds and pharmaceutical compositions for treating HCV, e.g., by inhibiting HCV NS5b polymerase,

In one embodiment, the present application provides for compounds having a structure according to Formula I, Formula II, Formula III or Formula IV:

W ² ' _R 3a-^C 7^ _R 8 W ^{2 L} R ^3b ^\ r^- R ⁸

R ⁴ R ^{7 R4 i i R7}

OR ^a OR ^a OR ^a OR ^a

Formula I Formula II

£>,

W ² -L ² V V B ^*2

W ^{2 R3d \ / ~ Rβ R F\P O PR a~R7 R R\ 6 O PR a"R7}

20 Formula HI Formula IV or a pharmaceutically acceptable salt, solvate, and/or ester thereof, wherein:

U iS -O-, -S-, or -N(R")-;

2

5 L ² is -C(R ¹⁰ J ₂ -; each R ^3a is CH2R ⁹ , alkenyl, substituted alkenyl, alkynyl, or substituted alkynyl; each R ^3b is CH2R ⁹ , alkenyl, substituted alkenyl, alkynyl, or substituted alkynyl wherein R ⁹ is not H; 10 each R ³ * is CH2R ⁹ , alkenyl, substituted alkenyl, alkynyl, or substituted alkynyl wherein R ⁹ is not H, OH, or F; each R ^3d is H, CH2R ⁹ , alkenyl, substituted alkenyl, alkynyl, or substituted alkynyl; each R ⁴ is independently H, CH2R ⁹ , alkenyl, substituted alkenyl, alkynyl, or 15 substituted alkynyl; each R ⁵ and R ⁶ is independently H, N(R ^a >, N3, CN, NO2, SR ^a , halogen, CHbR ⁹ , alkenyl, substituted alkenyl, alkynyl, or substituted alkynyl; or R ⁵ and R ⁶ taken together are =O, =NR ^b , or =CR ^c R ^d ; or R ⁵ and R ⁶ taken together with the carbon atom to which they are attached form a 3-7 membered 20 heterocyclic ring wherein one carbon atom in the heterocyclic ring can optionally be replaced with -O-, -S- or -NR ^a ~; each R ^a is independently H, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, or substituted alkynyl; each R ^b is independently H, alkyl, substituted alkyl, alkenyl, substituted 25 alkenyl, alkynyl, substituted alkynyl, or OR ^a ; each R ^c and R ^d are independently H, alkyl, substituted alkyl, alkenyl, . substituted alkenyl, alkynyl, substituted alkynyl, or halo; each R ⁷ is independently H, CH2R ⁹ , alkenyl, substituted alkenyl, alkynyl, or substituted alkynyl; 30 each R ⁸ is independently H, CH2R ⁹ , halo, alkyl, substituted alkyl, haloalkyl, -CN, -N3, alkenyl, substituted alkenyl, alkynyl, or substituted alkynyl;

3

each R ⁹ is independently H, OH, halo, Na, CN, N(R ^a )2, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, or substituted alkynyl, wherein one or more of the non-adjacent carbon atoms in the alkyl or substituted alkyl is optionally replaced with -O-, -S- or -NR ^a -; each R ¹⁰ is independently H, alkoxy, alkyl, or halo;

10 each R ¹¹ is independently H, alkyl, aryl, or substituted aryl;

B ¹ is a nucleobase selected from

N

?15

< and

_R 1β

each R ¹³ is independently OH or NH2;

15 R ¹⁴ is H or CH ₃ ;

R ¹⁵ is H, amino, or halo;

R ¹⁶ is H, halo, OR ^17a , N(R ²⁰ )(R ²¹ ), N(R ²⁸ )N(R ²⁸ )S(O) ₂ R ²⁸ , alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, S(O) _m R ²⁸ , or

20 S(O) ₂ NR ^17a R ^17b , NR ^17a R ^17b , N ₃ , NO, Nθ2, formyl, cyano, -C(O)NR ^17a R ^17b , - C(S)NR ^17a R ^17b , or -C(O)OR ¹⁷³ ; each R ^17a and R ^17b are independently H, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl, or optionally substituted alkanoyl;

25

4

5 R ²⁰ is H or OR ^17a ;

R ²¹ is H, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, optionally substituted aryl, cycloalkyl, or arylalkyl; or

R ²⁰ and R ²¹ together with the nitrogen to which they are attached form an 10 optionally substituted 3-7 membered heterocyclic ring wherein one carbon atom of the heterocyclic ring can optionally be replaced with -O-, - S- or -NR ^a -;

E ² is >N, >C-R ²⁵ or >C-R ³⁰ ;

D ₇ E, and F ^x are each independently >N or XI-R ²⁵ ;

15 each R ²⁵ is independently H, cyano, nitro, azido, optionally substituted (Ci-

Cβjalkyl, optionally substituted (Ci-C^alkenyl, optionally substituted (G- C ₆ )alkynyl, -NHCONH ₂₇ C(O)NR ²⁶ R ²⁷ , C(S)NR ²⁶ R ²⁷ , C(O)OR ²⁸ , hydroxy, OR ²⁸ , S(O) _m R ²⁸ , S(O)mNR ²⁶ R ²⁷ , -NR ²⁶ R ²⁷ , halo, l,3-oxazol-2-yl, l,3-oxazol-5- yl, l,3-thiazol-2-yl, imidazol-2-yl, 2-oxo-[l,3]dithiol-4-yl, furan-2-yl, or 20 2H-[l,2,3]triazol-4-yl; each R ²⁶ and R ²⁷ is independently H, optionally substituted (Ci-Cβjalkyl, optionally substituted (Ci-C6)alkenyl, optionally substituted (Ci- Cδjalkynyl, optionally substituted (C3-Cβ)cycloalkyl, aryl, heteroaryl, optionally substituted heterocycle, hydroxy, optionally substituted (G- 25 Cβ)alkoxy; or R ²⁶ and R ²⁷ together with the nitrogen to which they are attached form an optionally substituted 3-7 membered heterocyclic ring wherein one carbon atom of the heterocyclic ring can optionally be replaced with -O-, -S- or -NR ^a -; each R ²⁸ IS independently H, optionally substituted (Ci -Cβ) alkyl, optionally 30 substituted (Ci-C6)alkenyl, optionally substituted (Ci-Ce)alkynyl, optionally substituted (Gs-Csjcycloalkyl, aryl, heteroaryl or heterocycle;

5 R ³⁰ is -O CR ³¹ , -CH=CHR ³² , formyl, -CH=NNHR ³³ , -CH=N(OR ³³ ), -CH(OR ³⁴ ),

Or-B(OR ³³ );

R ³¹ is H, tri(G-C6)alkylsilyl, (G-C ₆ )alkoxy(Ci-C6)alkyl, optionally substituted heteroaryl, optionally substituted aryl, carboxy, or (Ci-Cβjalkoxycarbonyl; R ³² is hydrogen or (Ci-Ce)alkoxy; 10 R ³³ is H or (G-C6)alkyl;

R ³⁴ is (Ci-Ce)alkyl; m is 0, 1, or 2; wherein each aryl or heteroaryl of R ²⁶ , R ²⁷ , R ²⁸ and R ³¹ is independently optionally substituted with one or more (Ci-C6)alkyl, (Ci-Cβjalkoxy, (Ci- 15 C6)alkanoyl, (Ci-C6)alkanoyloxy, (Ci-C6)alkoxycarbonyl,

NR ³⁵ R ³⁶ , -Cf=O)NR ³⁵ R ³⁶ , cyano, halo, hydroxy, nitro, carboxy, (C ₃ - Cβjcycloalkyl, (C3-C8)cycloalkoxy, guanidino, trifluoromethoxy, mercapto, S(O)mR ³⁸ , S(O) _m NR ³⁵ R ³⁶ or trifluoromethyl;

R ³⁵ and R ³⁶ are each independently H, (Ci-Cs)alkyl or (Ci-Cβ)alkanoyl; 20 R ³⁸ is H, (G-C ₆ )alkyl or (Ci-C6)alkanoyl;

B ² is a nucleobase selected from

_R 16

= ¹ N" I ^{* " D}

R 44 R ⁴¹

R ,42

_R 43_γ

N

R ⁴³ -X R ⁴³ -Y,

_R 55 _R 55

R ⁵ S R ⁵ ^ .N, .R ⁵⁸ N' ^• N

,f\r\r .-} J. _

^■ X

_R 60

R™

N

R ^{62 ^} N

_R 90

91

γ4 z ¹ - ^■ \

X ⁸ X ⁸

\ .W*

W ⁶ . Wl .wϋ

W ⁶ .'-" ^* . N /' W ^6' .-"" ^> . N ₇ u // l\ ■we

N-

/ O and /

E ¹ is >N or >C-R ^2S ;

E ² , R ¹⁶ , E, F* , and D are defined as for B ¹ ;

5 R ⁴⁰ is H, NR ^4a R ^4b , NHC(O)R ^4b , (Ci-Ce)alkylNR ^4a R ^4b , NHNH ₂ , cyano, (G-

Ca)HIkYl (C2-C6)alkenyl, (C2-Cδ)alkynyl, aryl(G-C6)alkyl, heterocycle(Ci- Cό)alkyl _/ halo, (Ci-C6)alkylthio, (Ci-Ce)BIkOXy, hydroxy, or mercapto; R ⁴¹ is H, (Ci-Ce)alkyl, hydroxy(Ci-Ce)alkyl, (C2-Cβ)alkenyl, (C2-C ₆ )alkynyl, heterocycle, aryl, aryl(Ci-Cβ)alkyl; 10 each R ⁴² is independently H, hydroxy, mercapto, cyano, -SNR^R* ¹ , -C(NH)NR^R* ¹ , -C(=NH)NHOH, -C(NH)NHOR ^4c , -C(= NH)NHNR ⁴ =R ^4d , NHCOR ⁴ =, SR ^4c , OR ⁴ "=, SOR ⁴ =, SO ₂ R ⁴ =, -C(O)NR ⁴ =R ^4d , -C(S)NR ⁴¹ R ⁴ ", or R ^4c ;

R ⁴³ is H, hydroxy, NR ^4c R ^4d , NHC(O)NR ⁴ =, NHNHR ⁴ =, (Ci-C6)alkyl, (C2- 15 Ce)alkenyl, (C2-Cδ)alkynyl, heterocycle, aryl, aryl(Ci-Ce)aIkyl, halo,

C(O)OR ⁴ =, C(O)NR ^4c R ^4d , or absent when Y is N; R ^4a and R ^4b are each independently hydrogen, (Ci-Cβjalkyl, (Cz-Ce) alkenyl,

(C∑-Cβjalkynyl, heterocycle, or aryl;

R ⁴ =, and R ^4d are each independently hydrogen, (Ci-Cό)alkyl, (C2-C6)alkenyl, 20 (C2-C6)alkynyl, heterocycle, or aryl;

X, Y, and W are each independently N, C, CR ^4c , S or P; each R ⁴⁴ and R ⁴⁵ is independently H, hydroxy, mercapto, cyano, -SNR ^4c R ^4d , -C(NH)NR ^t =R ⁴ ", -C(=NH)NHOH, -C(NH)NHOR4 _C , -C(= NH)NHNR ⁴ =R ^4d , NHCOR ⁴ =, SR ⁴ =, OR ⁴ =, SOR ⁴ =, 25 SO ₂ R ⁴ =, -C(O)NR ^4c R ^4d , -C(S)NR ⁴⁰ R^, or R ⁴ =;

R ⁴⁶ , and R ⁴⁷ together with the atoms to which they are attached form a heterocyclic ring; U is S or O; wherein each aryl or heterocycle of R ⁴⁰ , R ⁴¹ , R ⁴² , R ⁴³ , R ^4a , R ^4b , R ^4c , R ^4d , R ⁴⁴ and 30 R ⁴⁵ is optionally substituted with one or more (Ci-Cδ)alkyl, (Ci-

Ce)alkylthio, (Ci-C6)alkoxy, (Ci-Cejalkanoyl, (Ci-C6)alkanoyloxy, (Ci-

10

5 Cβjalkoxycarbonyl, cyano, halo, hydroxy, nitro, carboxy, (C3-C8)cycloalkyl,

(C3-C8)cycloalkoxy, trifluoromethoxy, mercapto, or trifluoromethyl; R ⁵⁰ is NR ^5a R ^sb , ONR ⁵³ R ⁵¹³ , NR ^5a NR ^5a R ^5b , SR ^5b , OR ^5b , H, hydroxy, (C ₁ -C6)alkyl,

(Ci-Cδ)alkenyl, (Ci-C6)alkynyl, or aryl; R ⁵¹ is (G-C6)alkyl, (G-Gs)alkanoyl, or aryl; 10 R ⁵⁵ is NR ⁵³ R ⁵ ", ONR ^5a R ^5b , NR ^5a NR ^5a R ^5b , SR ^5b , OR ^5b , H, halo, hydroxy, (G-

Ce)alkyl, (Ca-Ce) alkenyl, (C2-G>)alkynyl, or aryl; R ⁵⁶ is H, halo, (G-C ₆ )alkyl, (G-C ₆ )alkenyl or (Q-GOalkynyl;

R ⁵⁷ and R ⁵⁸ are each independently -L-R ⁵ *=; each L is independently a direct bond, -N(R ^5a )-, O or S; 15 each R ^5a and R ^5b is independently H, hydroxy, (Ci-C6)alkyl, (C2-Cό)alkenyl,

(C2-C6)alkynyl, or aryl; each R ⁵ = is NR ^5a R ^5b , H, hydroxy, (Ci-Cβ)alkyl, (C2-Ce)alkenyl, (C2-Cβ)alkynyl, or aryl; wherein each (Ci-C6)alkyl, (G-Cejalkenyl, (C2-Cδ)alkynyl, or aryl of R ⁵⁰ , R ⁵¹ , 20 R ⁵⁵ , R ⁵⁶ , R ⁵⁷ , R ⁵⁸ , R ^5a , R ^5b and R ^5c is optionally substituted with one or more

(G-C ₆ )alkyl, (Ci-C6)alkylthio, (Ci-C ₆ )alkoxy, (Ci-Cβ)alkanoyl, (G- Cβjalkanoyloxy, (G-C6)alkoxycarbonyl, cyano, halo, hydroxy, nitro, carboxy, (C3-C8)cycloalkyl, (C3-C8)cycloalkoxy, trifluoromethoxy, mercapto, or trifluoromethyl;

25 R ⁶⁰ , R ⁶¹ , and R ⁶² are each independently H, halo, NR ^6b R ^6c , hydroxyamino,

NR ^6b NR ^6b R ^6c , N ₃ , NO, NO ₂ , formyl, cyano, -C(O)NR^R ⁶ -, -C(S)NR ⁶¹¹ R ⁶ =, -C(O)OR ^6b , R ^6b , OR ^6b , or SR ^6b ; R ^6b , and R ^6c are each independently H, (G-C ₆ )alkyl, (G-Cejalkenyl, (C2-

Ce)alkynyl, aryl, (G-Cβjalkanoyl, or aryl(G-C6)alkyl;

30 wherein each (G-Cejalkyl, (C2-Ce)alkenyl, (C ₂ -C6)alkynyl, or aryl of R ^6b and

R ^6c is optionally substituted with one or more (G-G;)alkyl, (G-

11

5 Cβjalkylthio, (Ci-Cβjalkoxy, (Ci-Cejalkanoyl, (Ci-C6)alkanoyloxy, (Ci-

Ce)alkoxycarbonyl, cyano, halo, hydroxy, nitro, carboxy, (C3-Cβ)cycloalkyl, (C3-C8)cycloalkoxy, trifluoromethoxy, mercapto, or trifluoromethyl; X ⁵ , X ⁶ , and X ⁷ are each independently N, CH, or C-R ^7a ; R ⁷⁰ and R ^7a are each independently H, halo, NR ⁷¹⁵ R ⁷ *, hydroxyamino, 10 NR ⁷ WR^R ⁷ -=, Ns, NO, NO ₂ , formyl, cyano, -C(O)NR ⁷¹⁵ R ⁷ ', -C(S)NR ^7b R ⁷ ?, -C(O)OR ⁷ ", R ⁷ ", OR ⁷ ", or SR ^7b ; R ⁷ ", and R ^7c are each independently H, (Ci-C ₆ )alkyl, (G∑-Cejalkenyl, (C ₂ -

Cβjalkynyl, aryl, (Ci-Cβjalkanoyl, or aryl(Ci-C6)alkyl; A ⁸⁰ , B ⁸⁰ , and Y ⁸⁰ , are each independently H, halo, OR ⁸⁰ , S(O) _n R ⁸⁰ , NR ⁸⁰ R ⁸¹ , 15 cyano, trifluoromethyl, C(W ^8O )OR ⁸⁰ , C(W ⁸⁰ )SR ⁸⁰ , C (W ⁸⁰ )NR ⁸⁰ R ⁸¹ , nitro, azido, carbocyclic, (Ci-Cό)alkyl, (C2-Cό)alkenyl, (C2-Cό)alkynyl, aryl, aryl(Ci-C6)alkyl, or heterocycle; or A ⁸⁰ and B ⁸⁰ taken together with the carbon atoms to which they are attached from a 4-7 membered carbocyclic or heterocyclic ring; 20 W ⁸⁰ is O, S, NR ⁸⁰ ; n is 0, 1, or 2;

Z ⁸⁰ is O, S, NR ⁸⁰ , or CR ⁸⁰ R ⁸¹ ; each V ⁸⁰ is independently N or CR ⁸⁰ ; each R ⁸⁰ and R ⁸¹ is independently H, carbocycle, (Ci-C6)alkyl, (C2-C6)alkenyl, 25 (C2-Cό)alkynyl, halo, (Ci-C6)alkoxy, amino, methylamino, dimethylamino, cyano, (Ci-C6)alkanoyl, aryl, aryl(Ci-Cό)alkyl, an amino acid residue or heterocycle; or R ⁸⁰ and R ⁸¹ taken together with the atom(s) to which they are attached form a 3-7 membered carbocyclic or heterocyclic ring; ^' X ⁹ is CR ^90a or N; 30 X ¹⁰ is O, S, or NR ^91a ;

12

5 R ⁹⁰ and R ⁹¹ are each independently H, halo, hydroxy, (Ci-C6)alkoxy, NR ⁹ ° ^b R ^91b , aryl, heterocycle; (Ci-C6)alkyl, (C ₂ -Ce) alkenyl, (C ₂ -C ₆ ) alkynyl, (G- Cβ)cycloalkyl, aryl(Ci-Ce)alkyl, S(O) _m R ^90b , S(O)m(aryl), or S(O)mNR ^90b R ^91b ; R ⁹⁰⁸ is H, halo, methyl, azido, or amino; R ^91a is H, (Ci-C6)alkyl or (G-Ce) alkanoyl; 10 R ^90b and R ^91b are each independently H, (Ci-Cejalkyl, (Q-Csjcycloalkyl, aryl(Ci-C6)alkyl, aryl, (Ci-Ce) alkanoyl, or aryl-C(O)-; wherein each (Ci-Ce)alkyl, (Cs-Csjcycloalkyl, aryl(Ci-Ce)alkyl, aryl, (Ci-Ce) alkanoyl, aryl-C(O)- and heterocycle of R ⁹⁰ , R ⁹¹ , R ^91a , R ^90b and R ^91b are optionally substituted with one to four halo, hydroxy, amino, (Ci-C6)alkyl, 15 and (Ci-Ce)alkoxy; each Z ¹ is independently N, C-R ^9a , O, S, NR ^9b , >C=O, >C=S, >C=NR ^9b , >S=O, >S(O)2 or CH-R ^9a ; provided that if a Z ¹ participates in an optional bond represented by a dotted line — in the formula, then that Z ¹ is N or C-R ^9a ; and provided that if a Z ¹ does not participate in an optional bond 20 represented by a dotted line — in the formula, then that Z ¹ is O, S, NR ^9b ,

>C=O, >C=S, >C=NR ^9b , >S=O, >S(O)2 or CH-R ^9a ; X ⁸ is O, S, SO, SO ₂ , Se, SeO, SeO ₂ or NR ^9b ; each W ⁶ is C, CH, or N; wherein if a W ⁶ participates in an optional bond represented by a dotted line — in the formula, then that W ⁵ is C; and if a 25 W ⁶ does not participate in an optional bond represented by a dotted line —

- in the formula, then that W ⁶ is CH, or N; each R ^9a is independently H, halo, NR ^9c R ^9d , hydroxyamino, NR ^9c NR ^9c R ^9d , Ns, cyano, -C(O)NR ^9c R ^9d , -C(S)NR ^9c R ^9d , -C(S)NR ^9c R ^9d , -C(=NH)OR ^9c , R ^9c , OR ^9c , or SR ^9c ;

30 each R ^9b is independently H, (Ci-Ce)alkyl, (Ca-Ce) alkenyl, (C2-Ce)alkynyl, aryl,

(Ci-Ce)alkanoyl, or aryl(Ci-Ce)alkyl;

13

R ^9c and R ^9d are each independently H ₇ (Ci-C.6)alkyl, (C∑-Cejalkenyl, (C2-

Ce)alkynyl, aryl, (G-C6)alkanoyl, or aryl(G-C6)alkyl; Y3=Y ⁴ is -N=N-, -CH=N-, -N=CR ⁸³ -, or -CH=CR ⁸³ -; each R ^8a is independently H, halo, or (G~C6)alkyl; B ³ is a nucleobase selected from

_R 7Q

R ¹⁶

E N

-N

\

10 N D' and

W ¹ and W ² are each independently a group of the formula:

Y ¹

R ^{x /} -γ2 i p. ^\

Ϋ ²

R ^x

M2 wherein: each Y ¹ is independently O, S, NR, ⁺ N(O)(R), N(OR), ⁺ N(O)(OR), or

15 N-NR2; ^' . ^' each Y ² is independently a bond, O, CR ₂ , NR, ⁺ N(O)(R), N(OR), ⁺ N(O)(OR),

N-NR ₂ , S, S-S, S(O), or S(O) ₂ ; M2 is 0, 1 or 2; each R ^x is independently R ^y , a protecting group, or the formula:

20 M1d

14

5 wherein:

MIa, MIc, and Mid are independently 0 or 1; M12c is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12; or when taken together, two R ^x are optionally substituted C2-C4 alkylene thereby forming a phosphorous-containing heterocycle;

10 each R ^y is independently H, F, Cl, Br, I, OH, R, -CC=Y ¹ JR, -CC=Y ¹ JOR, -

CC=Y ¹ JN(R) ₂ , -N(R) ₂ , - ⁺ N(R) ₃ , -SR, -S(O)R, -S(O) ₂ R, -S(O)(OR), - S(O) ₂ (OR), -OC(=Y ¹ )R, -OC(=Y ¹ )OR, -OQ=Y ¹ XN(R) ₂ ), -SC(=Y ¹ )R, - SC(=Y ¹ )OR, -SQ=Y ¹ XN(R) ₂ ), -N(R)C(=Y ¹ )R, -N(R)C(=Y ¹ )OR, or - N(R)C(=Y ¹ )N(R) ₂ , amino (-NH2), ammonium (-NHb ⁺ ), alkylamino,

15 dialkylamino, trialkylammonium, Ci-Cs alkyl, Ci-Cs alkylhalide, carboxylate, sulfate, sulfamate, sulfonate, Ci-Cs alkylsulfonate, Ci-Cs alkylamino, Ci-Cs alkylhydroxyl, Ci-Cs alkylthiol, alkylsulfone (-SO2R), sulfonamide (-SO ₂ NR ₂ ), alkylsulfoxide (-SOR), ester (-C(=O)OR), amido (-C(=O)NR ₂ ), nitrile (-CN), azido (-N ₃ ), nitro (-NO2), Ci-Cs

20 . alkoxy (-OR), Ci-Cs alkyl, Ci-Cs substituted alkyl, C ₂ -Cs alkenyl,

C2-C8 substituted alkenyl, C ₂ -Cs alkynyl, C ₂ -Cs substituted alkynyl, a protecting group, or W ³ ; or when taken together, two R ^y on the same carbon atom forms a carbocyclic ring of 3 to 7 carbon atoms; each R is independently H, halogen, Ci-Cs alkyl, Ci-Cs substituted alkyl,

25 C ₂ -Cs alkenyl, C ₂ -Cs substituted alkenyl, C ₂ -Cs alkynyl, C ₂ -Cs substituted alkynyl, C6-C20 aryl, Cβ-G-o substituted aryl, C2-C20 heterocycle, C2-C20 substituted heterocycle or a protecting group; W ³ is W ⁴ or W ⁵ ; W ⁴ is R, -C(Y ^J )R ^y , -C(Y ¹ JW ⁵ , -SO ₂ R ^y , or -SO ₂ W ⁵ ; and

30 W ⁵ is a carbocycle or a heterocycle wherein W ⁵ is independently substituted with 0 to 3 R ^y groups.

15

5 In another embodiment, the present application provides for a pharmaceutical composition comprising a compound of Formula I, Formula II, Formula III, or Formula IV; or a pharmaceutically acceptable salt, solvate, and/or ester thereof; and a pharmaceutically acceptable carrier or excipient.

In another embodiment, the present application provides for a 10 pharmaceutical composition comprising a compound of Formula I, Formula II, Formula III, or Formula IV; at least one additional therapeutic agent; and a pharmaceutically acceptable carrier or excipient.

In another embodiment, the present application provides for combination pharmaceutical agent comprising:

15 a) a first pharmaceutical composition comprising a compound of

Formula I, Formula II, Formula III, or Formula IV; or a pharmaceutically acceptable salt, solvate, or ester thereof; and b) a second pharmaceutical composition comprising at least one additional therapeutic agent selected from the group consisting of interferons, 20 ribavirin analogs, NS3 protease inhibitors, alpha-glucosidase 1 inhibitors, hepatoprotectants, non-nucleoside inhibitors of HCV, and other drugs for treating HCV.

In another embodiment, the present application provides for a method of inhibiting HCV polymerase, comprising contacting a cell infected with HCV 25 with an effective amount of a compound of Formula I, Formula II, Formula III, or Formula IV; or a pharmaceutically acceptable salts, solvate, and/or ester thereof.

In another embodiment, the present application provides for a method of inhibiting HCV polymerase, comprising contacting a cell infected with HCV . 30 with an effective amount of a compound of Formula I, Formula II, Formula III, or Formula IV; or a pharmaceutically acceptable salts, solvate, and/or ester thereof; and at least one additional therapeutic agent.

16

5 In another embodiment, the present application provides for a method of treating HCV in a patient, comprising administering to said patient a therapeutically effective amount of a compound of Formula I, Formula π, Formula III, or Formula IV; or a pharmaceutically acceptable salt, solvate, and/or ester thereof.

10 In another embodiment, the present application provides for a method of treating HCV in a patient, comprising administering to said patient a therapeutically effective amount of a compound of Formula I, Formula II, Formula III, or Formula IV; or a pharmaceutically acceptable salt, solvate, and/or ester thereof; and at least one additional therapeutic agent.

15

DETAILED DESCRIPTION

Reference will now be made in detail to certain claims of the invention, examples of which are illustrated in the accompanying structures and formulas. While the invention will be described in conjunction with the enumerated claims,

20 it will be understood that they are not intended to limit the invention to those claims. On the contrary, the invention is intended to cover all alternatives, modifications, and equivalents, which may be included within the scope of the present invention as defined by the claims.

25 Definitions

Unless stated otherwise, the following terms and phrases as used herein are intended to have the following meanings:

When trade names are used herein, applicants intend to independently include the tradename product and the active pharmaceutical ingredient(s) of 30 the tradename product.

As used herein, "a compound of the invention" or "a compound of formula (I)" means a compound of formula (I) or a pharmaceutically acceptable

17

5 salt, solvate, or physiologically functional derivative thereof. Similarly, with respect to isolatable intermediates, the phrase "a compound of formula (number)" means a compound of that formula and pharmaceutically acceptable salts, solvates and physiologically functional derivatives thereof.

"Alkyl" is hydrocarbon containing normal, secondary, tertiary or cyclic

10 carbon atoms. For example, an alkyl group can have 1 to 20 carbon atoms (Le, C1-C20 alkyl), 1 to 10 carbon atoms (i.e., C1-C10 alkyl), or 1 to 6 carbon atoms (i.e., Ci-Ce alkyl). Examples of suitable alkyl groups include, but are not limited to, methyl (Me, -CH ₃ ), ethyl (Et, -CH2CH3), 1-propyl (n-Pr, n-propyl, -CH ₂ CHaCH ₃ ), 2-propyl (i-Pr, i-propyl, -CH(CHs) ₂ ), 1-butyl (n-Bu, n-butyl, -CH ₂ CH ₂ CH ₂ CH ₃ ),

15 2-methyl-l -propyl (i-Bu, i-butyl, -CH ₂ CH(CHa) ₂ ), 2-butyl (s-Bu, s- butyl, -CH(CH ₃ )CH ₂ CH ₃ ), 2-methyl-2-proρyl (t-Bu, t-butyl, -C(CHs) ₃ ), 1-pentyl (n-pentyl, -CH ₂ CH ₂ CH ₂ CH ₂ CH ₃ ), 2-pentyl (-CH(CH ₃ )CH ₂ CH ₂ CH ₃ ), 3-pentyl (-CH(CH ₂ CHs) ₂ ), 2-methyl-2-butyl (-C(CHs) ₂ CH ₂ CH ₃ ), 3-methyl-2-butyl (-CH(CH ₃ )CH(CHs) ₂ ), 3-methyl-l-butyl (-CH ₂ CH ₂ CH(CHs) ₂ ), 2-methyl-l-butyl

20 (-CH ₂ CH(CH ₃ )CH ₂ CH ₃ ), 1-hexyl (-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CHS), 2-hexyl

(-CH(CH ₃ )CH ₂ CH ₂ CH ₂ CHS), 3-hexyl (-CH(CH ₂ CHS)(CH ₂ CH ₂ CHS)), 2-methyl-2- pentyl (-C(CHa) ₂ CH ₂ CH ₂ CH ₃ ), 3-methyl-2-pentyl (-CH(CH ₃ )CH(CHS)CH ₂ CH ₃ ), 4-methyl-2-pentyl (-CH(CHs)CH ₂ CH(CHa) ₂ ), 3-methyl-3-pentyl (- C(CH ₃ )(CH ₂ CHs) ₂ ), 2-methyl-3-pentyl (-CH(CH ₂ CHS)CH(CHS) ₂ ), 2,3-dimethyl-2-

25 butyl (-C(CHs) ₂ CH(CHs) ₂ ), 3,3-dimethyl-2-butyl (-CH(CHs)C(CHs) ₃ , and octyl (-(CH ₂ J ₇ CHa).

"Alkoxy" means a group having the formula -O-alkyl, in which an alkyl group, as defined above, is attached to the parent molecule via an oxygen atom. The alkyl portion of an alkoxy group can have 1 to 20 carbon atoms (i.e., Ci-Go

30 alkoxy), 1 to 12 carbon atoms(z.e., Ci-Ci ₂ alkoxy), or 1 to 6 carbon atoms(i.e., Ci-Cβ alkoxy). Examples of suitable alkoxy groups include, but are not limited to, methoxy (-O-CHs or -OMe), ethoxy (-OCH ₂ CHs or -OEt), t-butoxy (-O-C(CHs)a

18

5 or -OtBu) and the like.

"Haloalkyl" is an alkyl group, as defined above, in which one or more hydrogen atoms of the alkyl group is replaced with a halogen atom. The alkyl portion of a haloalkyl group can have 1 to 20 carbon atoms (i.e., G.-C20 haloalkyl), 1 to 12 carbon atoms(z.e., G-C12 haloalkyl), or 1 to 6 carbon atoms(z._?., Ci-Gs alkyl).

10 Examples of suitable haloalkyl groups include, but are not limited to, -CF ₃ , -CHF ₂ , -CFH ₂ , -CH2CF3, and the like.

"Alkenyl" is a hydrocarbon containing normal, secondary, tertiary or cyclic carbon atoms with at least one site of unsaturation, i.e. a carbon-carbon, sp ² double bond. For example, an alkenyl group can have 2 to 20 carbon atoms (i.e.,

15 C2-C20 alkenyl), 2 to 12 carbon atoms (i.e., G.-Ci2 alkenyl), or 2 to 6 carbon atoms (i.e., C2-C6 alkenyl). Examples of suitable alkenyl groups include, but are not limited to, ethylene or vinyl (-CH=CHz), allyl (-CHaCH=CKb), cyclopentenyl (-CsH ₇ ), and 5-hexenyl (-CH ₂ CH ₂ CH ₂ CH ₂ CH=CH ₂ ).

"Alkynyl" is a hydrocarbon containing normal, secondary, tertiary or

20 cyclic carbon atoms with at least one site of unsaturation, i.e. a carbon-carbon, sp triple bond. For example, an alkynyl group can have 2 to 20 carbon atoms (i.e., C2-C20 alkynyl), 2 to 12 carbon atoms (i.e., C2-C12 alkyne,), or 2 to 6 carbon atoms (i.e., C2-C6 alkynyl). Examples of suitable alkynyl groups include, but are not limited to, acetylenic (-C≡CH), propargyl (-CH ₂ CsCH), and the like.

25 "Alkylene" refers to a saturated, branched or straight chain or cyclic hydrocarbon radical having two monovalent radical centers derived by the removal of two hydrogen atoms from the same or two different carbon atoms of a parent alkane. For example, an alkylene group can have 1 to 20 carbon atoms, 1 to 10 carbon atoms, or 1 to 6 carbon atoms. Typical alkylene radicals include, but are

30 not limited to, methylene (-CH2-), 1,1-ethyl (-CH(CH ₃ )-), 1,2-ethyl (-CH2CH2-), 1,1- propyl (-CH(CH ₂ CH ₃ )-), 1,2-propyl (-CH ₂ CH(CH ₃ )-), 1,3-propyl (-CH ₂ CH ₂ CH ₂ -), 1,4-butyl (-CH ₂ CH ₂ CH ₂ CH ₂ -), and the like.

19

5 "Alkenylene" refers to an unsaturated, branched or straight chain or cyclic hydrocarbon radical having two monovalent radical centers derived by the removal of two hydrogen atoms from the same or two different carbon atoms of a parent alkene. For example, and alkenylene group can have 1 to 20 carbon atoms, 1 to 10 carbon atoms, or 1 to 6 carbon atoms. Typical alkenylene radicals include,

10 but are not limited to, 1,2-ethylene (-CH=CH-).

" Alkynylene" refers to an unsaturated, branched or straight chain or cyclic hydrocarbon radical having two monovalent radical centers derived by the removal of two hydrogen atoms from the same or two different carbon atoms of a parent alkyne. For example, an alkynylene group can have 1 to 20 carbon atoms, 1

15 to 10 carbon atoms, or 1 to 6 carbon atoms. Typical alkynylene radicals include, but are not limited to, acetylene (-C≡C-), propargyl (-CEbC≡≡C-), and 4-pentynyl (-CH ₂ CH ₂ CH ₂ C=CH-).

"Amino" refers generally to a nitrogen radical which can be considered a derivative of ammonia, having the formula -N(X)2, where each "X" is

20 independently H, substituted or unsubstituted alkyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, etc. The hybridization of the nitrogen is approximately sp ^s . Nonlimiting types of amino include -NH ₂ , - N(alkyl) ₂ , -NH(alkyl), -N(carbocyclyl)2, -NH(carbocyclyl), -N(heterocyclyl) ₂ , - NH(heterocyclyl), -N(aryl) ₂ , -NH(aryl), -N(alkyl)(aryl), -N(alkyl) (heterocyclyl), -

25 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. Nonlirniting examples of amino groups include -NHz, -NH(CHs), -N(CH3)2, -NH(CH ₂ CH ₃ X - N(CH ₂ CHS) ₂ , -NH(phenyl), -N(phenyl) _2/ -NH(benzyl), -N(benzyl) ₂ , etc. Substituted alkylamino refers generally to alkylamino groups, as defined

30 above, in which at least one substituted alkyl, as defined herein, is attached to the amino nitrogen atom. Non-limiting examples of substituted alkylamino includes -

20

5 NH(alkylene-C(O)-OH), -NH(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 derived by the removal of one hydrogen atom from a single carbon atom of a parent aromatic ring system.

For example, an aryl group can have 6 to 20 carbon atoms, 6 to 14 carbon atoms, or 10 6 to 12 carbon atoms. Typical aryl groups include, but are not limited to, radicals derived from benzene (e.g., phenyl), substituted benzene, naphthalene, anthracene, biphenyl, and the like.

" Arylalkyl" 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 15 replaced with an aryl radical. Typical arylalkyl groups include, but are not limited to, benzyl, 2-ρhenylethan-l-yl, naphthylmethyl, 2-naphthylethan-l-yl, naphthobenzyl, 2-naphthophenylethan-l-yl and the like. The arylalkyl group can comprise 6 to 20 carbon atoms, e.g., the alkyl moiety is 1 to 6 carbon atoms and the aryl moiety is 6 to 14 carbon atoms. 20 " Arylalkenyl" 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, but also an sp ² carbon atom, is replaced with an aryl radical. The aryl portion of the arylalkenyl can include, for example, any of the aryl groups disclosed herein, and the alkenyl portion of the arylalkenyl can include, for 25 example, any of the alkenyl groups disclosed herein. The arylalkenyl group can comprise 6 to 20 carbon atoms, e.g., the alkenyl moiety is 1 to 6 carbon atoms and the aryl moiety is 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 30 atom, but also an sp carbon atom, is replaced with an aryl radical. The aryl portion of the arylalkynyl can include, for example, any of the aryl groups disclosed herein, and the alkynyl portion of the arylalkynyl can include, for

21

5 example, any of the alkynyl groups disclosed herein. The arylalkynyl group can comprise 6 to 20 carbon atoms, e.g., the alkynyl moiety is 1 to 6 carbon atoms and the aryl moiety is 6 to 14 carbon atoms.

The term "substituted" in reference to alkyl, alkylene, aryl, arylalkyl, alkoxy, heterocyclyl, heteroaryl, carbocyclyl, etc. , for example, "substituted

10 alkyl", "substituted alkylene", "substituted aryl", "substituted arylalkyl",

"substituted heterocyclyl", and "substituted carbocyclyl" means alkyl, alkylene, aryl, arylalkyl, heterocyclyl, carbocyclyl respectively, in -which one or more hydrogen atoms are each independently replaced with a non-hydrogen substituent. Typical substituents include, but are not limited to, -X, -R, -O,

15 O, -OR, -SR, -S-, -NR ₂ , -N ⁺ R ₃ ,

=NR, -CX ₃ , -CN, -OCN, -SCN, -N=C=O, -NCS, -NO, -NO2,

=N ₂ , -N ₃ , -NHC(O)R, -C(=O)R, -C(=O)NRR -S(O) ₂ -, -S(O) ₂ OH, -S(O) ₂ R, -OS(O ) ₂ OR, -S(O) ₂ NR, -S(O)R, -OP(O)(OR) ₂ , -P(O)(OR) ₂ , -P(O)(O) ₂ , -P(O)(OH) ₂ , -P (O)(OR)(O-), -C(O)R, -C(O)X, -C(S)R, -C(O)OR, -C(O)O-, -C(S)OR, -C(O)SR, -C(

20 S)SR, -C(O)NRR, -C(S)NRR, -C(=NR)NRR, where each X is independently a halogen: F, Cl, Br, or I; and each R is independently H, alkyl, aryl, arylalkyl, a heterocycle, or a protecting group or prodrug moiety. Alkylene, alkenylene, and alkynylene groups may also be similarly substituted. Unless otherwise indicated, when the term "substituted" is used in conjunction with groups such as arylalkyl,

25 which have two or more moieties capable of substitution, the substituents can be attached to the aryl moiety, the alkyl moiety, or both.

The term "prodrug" as used herein refers to any compound that when administered to a biological system generates the drug substance, i.e., active ingredient, as a result of spontaneous chemical reaction(s), enzyme catalyzed

30 chemical reaction(s), photolysis, and/or metabolic chemical reaction(s). A prodrug is thus a covalently modified analog or latent form of a therapeutically active compound.

22

5 One skilled in the art will recognize that substituents and other moieties of the compounds of Formula I-IV should be selected in order to provide a compound "which is sufficiently stable to provide a pharmaceutically useful compound "which can be formulated into an acceptably stable pharmaceutical composition. Compounds of Formula I-IV which have such stability are

10 contemplated as falling "within the scope of the present invention.

"Heteroalkyl" refers to an alkyl group where one or more carbon atoms have been replaced with a heteroatom, such as, O, N, or S. For example, if the carbon atom of the alkyl group "which is attached to the parent molecule is replaced with a heteroatom (e.g., O, N, or S) the resulting heteroalkyl groups are,

15 respectively, an alkoxy group (e.g., -OCH3, etc.), an amine (e.g., -NHCH3, -N(OHb) ₂ , etc.), or a thioalkyl group (e.g., -SCH3). If a non-terminal carbon atom of the alkyl group which is not attached to the parent molecule is replaced with a heteroatom (e.g., O, N, or S) the resulting heteroalkyl groups are, respectively, an alkyl ether (e.g., -CH2CH2-O-CH3, etc.), an alkyl amine (e.g., -CH2NHCH3, -CH ₂ N(CHs) ₂ , etc.),

20 or a thioalkyl ether (e.g _v -CH2-S-CH3). If a terminal carbon atom of the alkyl group is replaced with a heteroatom (e.g., O, N, or S), the resulting heteroalkyl groups are, respectively, a hydroxyalkyl group (e.g., -CH ₂ CH ₂ -OH), an aminoalkyl group (e.g., -CH ₂ NH.), or an alkyl thiol group (e.g., -CH2CH2-SH). A heteroalkyl group can have, for example, 1 to 20 carbon atoms, 1 to 10 carbon atoms, or 1 to 6 carbon

25 atoms. A Ci-Cβ heteroalkyl group means a heteroalkyl group having 1 to 6 carbon atoms.

"Heterocycle" or "heterocyclyl" as used herein includes by way of example and not limitation those heterocycles described in Paquette, Leo A.; Principles of Modern Heterocyclic Chemistry (W.A. Benjamin, New York, 1968),

30 particularly 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), in particular Volumes 13, 14, 16, 19, and 28; and /. Am. Chem. Soc. (1960)

23

5 82:5566. In one specific embodiment of the invention "heterocycle" includes a "carbocycle" as defined herein, wherein one or more (e.g. 1, 2, 3, or 4) carbon atoms have been replaced -with a heteroatom (e.g. O, N, or S). The terms "heterocycle" or "heterocyclyl" includes saturated rings, partially unsaturated rings, and aromatic rings (i.e., heteroaromatic rings). Substituted heterocyclyls 10 __ include, for example, heterocyclic rings substituted with any of the substituents disclosed herein including carbonyl groups. A non-limiting example of a carbonyl substituted heterocyclyl is: n

^ T o

Examples of heterocycles include by way of example and not limitation

15 pyridyl, dihydroypyridyl, tetrahydropyridyl (piperidyl), thiazolyl, tetrahydrothiophenyl, sulfur oxidized tetrahydrothiophenyl, pyrimidinyl, furanyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, tetrazolyl, benzofuranyl, thianaphthalenyl, indolyl, indolenyl, quinolinyl, isoquinolinyl, benzimidazolyl, piperidinyl, 4-piperidonyl, pyrrolidinyl, 2-pyrrolidonyl, pyrrolinyl,

20 tetrahydrofuranyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, octahydroisoqujnolinyl, azocinyl, triazinyl, 6H-1,2,5- thiadiazinyl, 2H _/ 6H-l,5,2-dithiazinyl _/ thienyl, thianthrenyl, pyranyl, isobenzofuranyl, chromenyl, xanthenyl, phenoxathinyl, 2H-pyrrolyl, isothiazolyl, isoxazolyl, pyrazinyl, pyridazinyl, indolizinyl, isoindolyl, 3H-indolyl, IH-

25 indazoly, purinyl, 4H-quinolizinyl, phthalazinyl, naphthyridinyl, quinoxalinyl, quinazolinyl, cinnolinyl, pteridinyl, 4aH-carbazolyl, carbazolyl, β-carbolinyl, phenanthridinyl, acridinyl, pyrimidinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, furazanyl, phenoxazinyl, isochromanyl, chromanyl, imidazolidinyl, imidazolinyl, pyrazolidinyl, pyrazolinyl, piperazinyl, indolinyl, .

30 isoindolinyl, quinuclidinyl, morpholinyl, oxazolidinyl, benzotriazolyl,

24

5 benzisoxazolyl, oxindolyl, benzoxazolinyl, isatinoyl, and bis-tetrahydro£uranyl:

O

By way of example and not limitation, carbon bonded heterocycles are bonded at position 2, 3, 4, 5, or 6 of a pyridine, position 3, 4, 5, or 6 of a pyridazine, position 2, 4, 5, or 6 of a pyrimidine, position 2, 3, 5, or 6 of a

10 pyrazine, position 2, 3, 4, or 5 of a furan, tetrahydrofuran, thiofuran, thiophene, pyrrole or tetrahydropyrrole, position 2, 4, or 5 of an oxazole, imidazole or thiazole, position 3, 4, or 5 of an isoxazole, pyrazole, or isothiazole, position 2 or 3 of an aziridine, position 2, 3, or 4 of an azetidine, position 2, 3, 4, 5, 6, 7, or 8 of a quinoline or position 1, 3, 4, 5, 6, 7, or 8 of an isoquinoline. Still more typically,

15 carbon bonded heterocycles include 2-pyridyl, 3-pyridyl, 4-pyridyl, 5-pyridyl, 6- pyridyl, 3-pyridazinyl, 4-pyridazinyl, 5-pyridazinyl, 6-pyridazinyl, 2- pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, 6-pyrimidinyl, 2-ρyrazinyl, 3- pyrazinyl, 5-pyrazinyl, 6-pyrazinyl, 2-thiazolyl, 4-thiazolyl, or 5-thiazolyl.

By way of example and not limitation, nitrogen bonded heterocycles are

20 bonded at position 1 of an aziridine, azetidine, pyrrole, pyrrolidine, 2-pyrroline, 3-pyrroline, imidazole, imidazolidine, 2-imidazoline, 3-imidazoline, pyrazole, pyrazoline, 2-pyrazoline, 3-pyrazoline, piperidine, piperazine, indole, indoline, lH-indazole, position 2 of a isoindole, or isoindoline, position 4 of a morpholine, and position 9 of a carbazole, or β-carboline. Still more typically, nitrogen

25 bonded heterocycles include 1-aziridyl, 1-azetedyl, 1-pyrrolyl, 1-imidazolyl, 1- pyrazolyl, and 1-piperidinyl.

"Heterocyclylalkyl" 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 with a heterocyclyl radical (i.e., a heterocyclyl-alkylene-

30 moiety). Typical heterocyclyl alkyl groups include, but are not limited to

25

5 heterocyclyl-CIHb-, 2-(heterocyclyl)ethan-l-yl, and the like, wherein the

"heterocyclyl" portion includes any of the heterocyclyl groups described above, including those described in Principles of Modern Heterocyclic Chemistry. One skilled in the art will also understand that the heterocyclyl group can be attached to the alkyl portion of the heterocyclyl alkyl by means of a carbon-carbon bond

10 or a carbon-heteroatom bond, with the proviso that the resulting group is chemically stable. The heterocyclyl alkyl group comprises 6 to 20 carbon atoms, e.g., the alkyl portion of the arylalkyl group is 1 to 6 carbon atoms and the heterocyclyl moiety is 5 to 14 carbon atoms. Examples of heterocyclylalkyls include by way of example and not limitation 5-membered sulfur, oxygen,

15 and/or nitrogen containing heterocycles such as thiazolylmethyl, 2- thiazolylethan-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.

20 "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, but also a sp ² carbon atom, is replaced with a heterocyclyl radical (i.e., a heterocyclyl-alkenylene- moiety). The heterocyclyl portion of the heterocyclyl alkenyl group includes any of the heterocyclyl groups described herein,

25 including those described in Principles of Modern Heterocyclic Chemistry, and the alkenyl portion of the heterocyclyl alkenyl group includes any of the alkenyl groups disclosed herein. One skilled in the art will also understand that the heterocyclyl group can be attached to the alkenyl portion of the heterocyclyl alkenyl by means of a carbon-carbon bond or a carbon-heteroatom bond, with

30 the proviso that the resulting group is chemically stable. The heterocyclyl alkenyl group comprises 6 to 20 carbon atoms, e.g., the alkenyl portion of the heterocyclyl alkenyl group is 1 to 6 carbon atoms and the heterocyclyl moiety is

26

5 5 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; but also an sp carbon atom, is replaced with a heterocyclyl radical (i.e., a heterocyclyl-alkynylene- moiety). The heterocyclyl portion of the heterocyclyl

10 alkynyl group includes any of the heterocyclyl groups described herein, including those described in Principles of Modern Heterocyclic Chemistry, and the alkynyl portion of the heterocyclyl alkynyl group includes any of the alkynyl groups disclosed herein. One skilled in the art will also understand that the heterocyclyl group can be attached to the alkynyl portion of the heterocyclyl

15 alkynyl by means of a carbon-carbon bond or a carbon-heteroatom bond, with the proviso that the resulting group is chemically stable. The heterocyclyl alkynyl group comprises 6 to 20 carbon atoms, e.g., the alkynyl portion of the heterocyclyl alkynyl group is 1 to 6 carbon atoms and the heterocyclyl moiety is 5 to 14 carbon atoms.

20 "Heteroaryl" refers to an aromatic heterocyclyl having at least one heteroatom in the ring. Non-limiting examples of suitable heteroatoms which can be included in the aromatic ring include oxygen, sulfur, and nitrogen. Non- limiting examples of heteroaryl rings include all of those listed in the definition of "heterocyclyl", including pyridinyl, pyrrolyl, oxazolyl, indolyl, isoindolyl,

25 purinyl, furanyl, thienyl, benzofuranyl, benzothiophenyl, carbazolyl, imidazolyl, thiazolyl, isoxazolyl, pyrazolyl, isothiazolyl, quinolyl, isoquinolyl, pyridazyl, pyrimidyl, pyrazyl, etc.

"Carbocycle" or "carbocyclyl" refers to a saturated (i.e., cycloalkyl), partially unsaturated (e.g., cycloakenyl, cycloalkadienyl, etc.) or aromatic ring

30 having 3 to 7 carbon atoms as a monocycle, 7 to 12 carbon atoms as a bicycle, and up to about 20 carbon atoms as a polycycle. Monocyclic carbocycles have 3 to 6 ring atoms, still more typically 5 or 6 ring atoms. Bicyclic carbocycles have 7

27

5 to 12 ring atoms, e.g., arranged as a bicyclo [4,5], [5,5], [5,6] or [6,6] system, or 9 or 10 ring atoms arranged as a bicyclo [5,6] or [6,6] system, or spiro-fused rings. Non-limiting examples of monocyclic carbocycles include cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-l-enyl, l-cyclopent-2-enyl, l-cyclopent-3- enyl, cyclohexyl, 1-cyclohex-l-enyl, l-cyclohex-2-enyl, l-cyclohex-3-enyl, and

10 phenyl. Non-limiting examples of bicyclo carbocycles includes naphthyl.

"Arylheteroalkyl" refers to a heteroalkyl as defined herein, in which a hydrogen atom (which may be attached either to a carbon atom or a heteroatom) has been replaced with an aryl group as defined herein. The aryl groups may be bonded to a carbon atom of the heteroalkyl group, or to a heteroatom of the

15 heteroalkyl group, provided that the resulting arylheteroalkyl group provides a chemically stable moiety. For example, an arylheteroalkyl group can have the general formulae -alkylene-

O-aryl, -alkylene-O-alkylene-aryl, -alkylene-NH-aryl, -alkylene-NH-alkylene-ary 1, -alkylene-S-aryl, -alkylene-S-alkylene-aryl, etc. In addition, any of the alkylene

20 moieties in the general formulae above can be further substituted with any of the substituents defined or exemplified herein.

"Heteroarylalkyl" refers to an alkyl group, as defined herein, in which a hydrogen atom has been replaced with a heteroaryl group as defined herein. Non-limiting examples of heteroaryl alkyl

25 include -Ctk-pyridinyl, -CH2-pyrrolyl, -CFfe-oxazolyl, -CHa-indolyl, -GHb-isoind ^■ plyl, -CH2-purinyl, -CH∑-furanyl, -CH2-thienyl, -CHi-benzofuranyl, -CH2-benzot hiophenyl, -CH2-carbazolyl, -CH2-imidazolyl, -CH2-thiazolyl, -QHb-isoxazolyl, -C H2-pyrazolyl, -CH2-isothiazolyl, -CHb-quinolyl, -CH_-isoquinolyl, -CHb-pyridazy 1, -CHa-pyrimidyl, -CHz-pyrazyl, -CH(CH3)-pyridinyl, -CH(CH3>-pyrrolyl, -CH(C

30 H3)-oxazolyl, -CH(CH ₃ )-indolyl, -CH(CH ₃ )-isoindolyl, -CH(CH ₃ )-purinyl, -CH(C H3)-furanyl, -CH(CH ₃ )-thienyl, -CH(CH ₃ ) -benzofuranyl, -CH(CHs)-benzothiophe nyl, -CH(CHs)-carbazolyl, -CH(CH3)-imidazolyl, -CH(CH3)-thiazolyl, -CH(CHs)-I

28

5 soxazolyl, -CH(CHs)-ρyrazolyl _/ -CH(CH ₃ )-isothiazolyl, -CH(CH3)-quinolyl, -CH( CH3)-isoquinolyl, -CH(CH ₃ )-pyridazyl _/ -CH(CH3)-pyrimidyl, -CH(CH3)-pyrazyI, etc.

The term "optionally substituted" in reference to a particular moiety of the compound of Formula I, II, III, or IV (e.g., an optionally substituted aryl 10 group) refers to a moiety having 0, 1, 2, or more substituents.

"Linker" or "link" means a chemical moiety comprising a covalent bond or a chain of atoms. Linkers include repeating units of alkyloxy (e.g. polyethyleneoxy, PEG, polymethyleneoxy) and alkylamino (e.g. polyethyleneamino, Jeff amine™); and diacid ester and amides including 15 succinate, succinamide, diglycolate, malonate, and caproamide.

The terms such as "oxygen-linked", "nitrogen-linked", "carbon-linked", "sulfur-linked", or "phosphorous-linked" mean that if a bond between two moieties can be formed by using more than one type of atom in a moiety, then the bond formed between the moieties is through the atom specified. For 20 example, a nitrogen-linked amino acid would be bonded through a nitrogen atom of the amino acid rather than through an oxygen or carbon atom of the amino acid.

Certain Y ¹ and Y ² alternatives are nitrogen oxides such as ⁺ N(O)(R) or ⁴ N(O)(OR). These nitrogen oxides, as shown here attached to a carbon atom, can

O ^"

25 also be represented by charge separated groups such as ^R or

^ ^OR , respectively, and are intended to be equivalent to the afore mentioned representations for the purposes of describing this invention.

29

Unless otherwise specified, the carbon atoms of this invention are intended to have a valence of four. In some chemical structure representations where carbon atoms do not have a sufficient number of variables attached to produce a valence of four, the remaining carbon substitutents needed to provide a valence of four should be assumed to be hydrogen. For example,

O

NH

10 ^Ύ R ^{2 T} Ύ has the same meaning as

CH ₃ HL ^L ^O

Whenever a compound described herein is substituted with more than one of the same designated group, e.g., "R" or "R ¹ ", then it will be understood that the groups may be the same or different, i.e., each group is independently 15 selected. Wavy lines indicate the site of covalent bond attachments to the adjoining substructures, groups, moieties, or atoms.

The term "chiral" refers to molecules which have the property of non- superimposability of the mirror image partner, while the term "achiral" refers to molecules which are superimposable on their mirror image partner. 20 The term "stereoisomers" refers to compounds which have identical chemical constitution, but differ with regard to the arrangement of the atoms or groups in space.

30

5 "Diastereomer" refers to a stereoisomer with two or more centers of chirality and whose molecules are not mirror images of one another. Diastereomers have different physical properties, e.g., melting points, boiling points, spectral properties, and reactivities. Mixtures of diastereomers may separate under high resolution analytical procedures such as electrophoresis and

10 chromatography.

"Enantiomers" refer to two stereoisomers of a compound which are non- superimposable mirror images of one another.

Stereochemical definitions and conventions used herein generally follow S. P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-

15 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 plane-polarized light. In describing an optically active compound, the prefixes D and L or R and S are used to denote the absolute configuration of the

20 molecule about its chiral center(s). The prefixes d and 1 or (+) and (-) are employed to designate the sign of rotation of plane-polarized light by the compound, with (-) or 1 meaning that the compound is levorotatory. A compound prefixed with (+) or d is dextrorotatory. For a given chemical structure, these stereoisomers are identical except that they are mirror images of

25 one another. A specific stereoisomer may also be referred to as an enantiomer, and a mixture of such isomers is often called an enantiomeric mixture. A 50:50 mixture of enantiomers is referred to as a racemic mixture or a racemate, which may occur where there has been no stereoselection or stereospecificity in a chemical reaction or process. The terms "racemic mixture" and "racemate" refer

30 to an equimolar mixture of two enantiomeric species, devoid of optical activity.

"Nucleobase" or "nucleoside base" means any nitrogen-containing

31

5 heterocyclic moiety capable of forming Watson-Crick hydrogen bonds in. pairing with a complementary nucleobase or nucleobase analog, e.g. a purine, a 7- deazapurine, or a pyrimidine. Typical nucleobases are the naturally-occurring nucleobases: adenine, guanine, cy tosine, uracil, thymine, and analogs of the naturally-occurring nucleobases, e.g. 7-deazaadenine, substituted 7-deazapurines

10 such as 7-alkynyl, 7-cyano, 7-carboxamido, 7-deazaguarrine, 7-deaza-8-azaguanine, 7-deaza-8-azaadenine, inosine, nebularine, nitropyrrole, nitroindole, 2- aminopurine, 2-amino-6-chloropurine, 2,6-diaminopurine, hypoxanthine, pseudouridine, pseudocytosine, pseudoisocytosine, 5-propynylcytosine, isocytosine, isoguanine, 7-deazaguanine, 2-thiopyrimidiαe, 6-thioguanine, 4-

15 thiothymine, 4-thiouradl, O ⁶ -methylguanine, N ⁶ -methyladenine, O- methylthymine, 5,6-dihydrothymine, 5,6-dihydrouracil, 4-methylindole, pyrazine bases including 3-oxo-2-carboxamidopyrazine, 5-fluoro-3-oxo-2- carboxamidopy r azine, 6-fluoro-3-oxo-2-carboxamidopy razine, py razolo [3, 4- D]pyrimidines (U.S. Patent Nos. 6,143,877 and 6,127,121; WO 01/38584), and

20 ethenoadenine (Fasman (1989) in Practical Handbook of Biochemistry and Molecular Biology, pp. 385-394, CRC Press, Boca Raton, Fl).

The invention provides compounds of Formula I, II, III or IV wherein B ¹ , B ² , or B ³ is a nucleoside base. The compounds may include any nucleoside base, provided the final compound possesses useful therapeutic (e.g. anti-viral)

25 properties. Additional nucleoside bases that can be incorporated into the compounds of this invention are disclosed in United States Patent Application Publication Number 2004/0147464, United States Patent Application Publication Number 2005/0215511, International Patent Application Publication Number WO 03/061385, International Patent Application Publication Number WO

30 03/062257, International Patent Application Publication Number WO 031 '072757, International Patent Application Publication Number WO 03/073989,

32

International Patent Application Publication Number WO 2005/021568, International Patent Application Publication Number WO 2005/123087, International Patent Application Publication Number WO 2006/002231, and International Patent Application Publication Number WO 2006/000922.

In one embodiment, compounds of this invention are of Formula I:

^W R ^3a -λ /^R ⁸

^R i I ^R?

10 OR ^a OR ^a

Formula I wherein all variables are defined as above for Formula I. In one aspect of this embodiment, each R ^a is H. In another aspect of this embodiment, R ^3a is CH2R ⁹ . In another aspect of this embodiment, R ^3a is alkenyl or substituted alkenyl. In 15 another aspect of this embodiment, R ^3a is alkynyl or substituted alkynyl. In another aspect of this embodiment, each R ¹⁰ is H and L ¹ is O.

In preferred embodiment of Formula I, B ¹ is R ¹³

. In one aspect of this embodiment, each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ^3a is CH2R ⁹ . In another 20 aspect of this embodiment, R ^3a is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3a is alkynyl or substituted alkynyl. In another aspect of this embodiment, R ^3a is CH2R ⁹ and R ⁹ is not H.

In another preferred embodiment of Formula I, B ¹ is

33

_R 13

N

/

wherein each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In one aspect of this embodiment, R ^3a is CH2R ⁹ . In another aspect of this embodiment, R ^3a is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3a is alkynyl or substituted alkynyl. In another aspect of this embodiment, R ^3a is CH2R ⁹ and R ⁹ is not H. In another aspect of this embodiment, R ¹³ is NH2. In

10 another aspect of this embodiment, R ¹³ is NH2 and R ¹⁵ is H. In another aspect of this embodiment, R ¹³ is OH. In another aspect of this embodiment, R ¹³ is OH and R ¹⁵ is NH2. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently

15 Y ² -R ^x wherein each Y ² is -O- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R ^x is not H.

In another preferred embodiment of Formula I, B ¹ is

R ^{13 ' •}

wherein R ¹³ is NH ₂ , each R ¹⁰ is H, L ¹ is O, and each R ^a is H.

20 In one aspect of this embodiment, R ^3a is CH2R ⁹ . In another aspect of this embodiment, R ^3a is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3a is alkynyl or substituted alkynyl. In another aspect of this embodiment, R ^3a is CH2R ⁹ and R ⁹ is not H. In another aspect of this embodiment, R ^3a is methyl. In another aspect of this embodiment, R ^3a is ethylenyl. In another

34

5 aspect of this embodiment, R ^3a is ethynyl. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R* wherein each Y ² is independently -O- or -N(R)- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is -O- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is 10 independently -N(R)- and R ^x is not H.

In another preferred embodiment of Formula I, B ¹ is R ¹³

wherein R ¹³ is OH, each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In one aspect of this embodiment, R ^3a is CH2R ⁹ . In another aspect of this embodiment, R ^3a is alkenyl or substituted alkenyl. In another aspect of this

15 embodiment, R ^3a is alkynyl or substituted alkynyl. In another aspect of this embodiment, R ^3a is CH2R ⁹ and R ⁹ is not H. In another aspect of this embodiment, R ^3a is methyl. In another aspect of this embodiment, R ^3a is ethylenyl. In another aspect of this embodiment, R ^3a is ethynyl. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or

20 -N(R)- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is -O- and R ^x is not H. In another aspect of this embodiment, each W and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R ^x is not H.

In another preferred embodiment of Formula I, B ¹ is

35

_R 13

R ¹ 4 N

T V. Ml

5 >nw . In one aspect of this embodiment, R ¹⁰ is H, L ¹ is O, and each

R ^a is H. In another aspect of this embodiment, R ^3a is CH2R ⁹ . In another aspect of this embodiment, R ^3a is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3a is alkynyl or substituted alkynyl. In another aspect of this embodiment, R ^3a is CH2R ⁹ and R ⁹ is not H.

10 In another preferred embodiment of Formula I, B ¹ is

_R 13

•"λ^ wherein each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In one aspect of this embodiment, R ^3a is CH2R ⁹ . In another aspect of this embodiment, R ^3a is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3a is alkynyl or substituted alkynyl. In another aspect of this embodiment, R ^3a is

15 CH2R ⁹ and R ⁹ is not H. In another aspect of this embodiment, R ¹³ is NH2. In another aspect of this embodiment, R ¹³ is NH2 and R ¹⁴ is H. In another aspect of . this embodiment, R ¹³ is OH. In another aspect of this embodiment, R ¹³ is OH and R ¹⁴ is H. . In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is -O- and R* is not H. In another aspect of

20 this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R ^x is not H.

In another preferred embodiment of Formula I, B ¹ is

36

5 >^ ^r w ^> wherein R ¹³ is NH ₂ , R ¹⁴ is H, each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In one aspect of this embodiment, R ^3a is CHzR ⁹ . In another aspect of this embodiment, R ^3a is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3a is alkynyl or substituted alkynyl. In another aspect of this embodiment, R ^3a is CH2R ⁹ and R ⁹ is not H. In another aspect of this embodiment,

10 R ^3a is methyl. In another aspect of this embodiment, R ^3a is ethylenyl. In another aspect of this embodiment, R ^3a is ethynyl. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R ^x is not H. In another aspect of this embodiment, each W ^: and W ² is independently Y ² -R ^x wherein each Y ² is -O- and R ^x is not H. In another aspect of

15 this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R ^x is not H.

In another preferred embodiment of Formula I, B ¹ is

R ¹³

^{I X λλλP wherein R 13 is OH, R 14 is H, each R 10 is H, L 1 is O, and each R a is H. In one aspect of this embodiment, R 3a is CH2R 9 . In another aspect of this 20 embodiment, R 3a is alkenyl or substituted alkenyl. In another aspect of this embodiment, R 3a is alkynyl or substituted alkynyl. In another aspect of this embodiment, R 3a is CH2R 9 and R 9 is not H. In another aspect of this embodiment, R 3a is methyl. In another aspect of this embodiment, R 3a is ethylenyl. In another aspect of this embodiment, R 3a is ethynyl. In another aspect of this embodiment,}

37

5 each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently — O- or -N(R)- and R* is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is -O- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R ^x is not H.

10 In another preferred embodiment of Formula I, B ¹ is

_R 16

. In one aspect of this embodiment, each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ^3a is CRbR ⁹ . In another aspect of this embodiment, R ^3a is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3a is alkynyl or substituted alkynyl. In another 15 aspect of this embodiment, R ^3a is CH2R ⁹ and R ⁹ is not H.

In another preferred embodiment of Formula I, B ¹ is

^" _R 1β

^V ^^ ^V wherein each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In one aspect of this embodiment, R ^3a is CH2R ⁹ . In another aspect of this embodiment, R ^3a is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3a is 20 alkynyl or substituted alkynyl. In another aspect of this embodiment, R ^3a is

CH2R ⁹ and R ⁹ is not H. In another aspect of this embodiment, each E and D is >N. In another aspect of this embodiment, each E and D is >N and F ^x is XZ-R ²⁵ .

In another preferred embodiment of Formula I, B ¹ is

38

_R 16

5 ^r wherein each E and D is >N, each R ¹⁰ is H, L ¹ is O, and each R ^a is

H. In one aspect of this embodiment, R ^3a is CH2R ⁹ . In another aspect of this embodiment, R ^3a is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3a is alkynyl or substituted alkynyl. In another aspect of this embodiment, R ^3a is CH2R ⁹ and R ⁹ is not H. In another aspect of this embodiment 10 F ^x is >C-R ²⁵ .

In another preferred embodiment of Formula I, B ¹ is

_R 1β

wherein each E and D is >N, F ^x is >C-R ²⁵ , each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In one aspect of this embodiment, R ^3a is CH2R ⁹ . In another aspect of this embodiment, R ^3a is alkenyl or substituted alkenyl. In another

15 aspect of this embodiment, R ^3a is alkynyl or substituted alkynyl. In another aspect of this embodiment, R ^3a is CH2R ⁹ and R ⁹ is not H. In another aspect of this embodiment, R ^3a is methyl. In another aspect of this embodiment, R ^3a is ethylenyl. In another aspect of this embodiment, R ^3a is ethynyl. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each

20 Y ² is independently — O- or -N(R)- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is -O- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R ^x is not H.

In another preferred embodiment of Formula I, B ¹ is

39.

_R 16

E ²

5 ^{N D} . In one aspect of this embodiment, each R ¹⁰ is H _x L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ^3a is CH2R ⁹ . In another aspect of this embodiment, R ^3a is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3a is alkynyl or substituted alkynyl. In another aspect of this embodiment, R ^3a is CH2R ⁹ and R ⁹ is not H. In another aspect of this 10 embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is -O- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R* is not H.

15 In one embodiment, compounds of this invention are of Formula II:

^W _R 3b^ Y— R ^β

^{R4 I 1 R7}

OR ^a OR ^a Formula II wherein all variables are defined as above for Formula II. In one aspect of this embodiment, each R ^a is H. In another aspect of this embodiment, R ^3b is 20 CH2R ⁹ wherein R ⁹ is not H. In another aspect of this embodiment, R ³¹ ? is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3b is alkynyl or substituted alkynyl. In another aspect of this embodiment, each R ¹⁰ is H and L ¹ is O.

In a preferred embodiment of Formula II,- B ² is

40

. In one aspect of this embodiment, each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ^3b is CH2R ⁹ wherein R ⁹ is not H. In another aspect of this embodiment, R ^3b is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3b is alkynyl or substituted alkynyl.

10 In another preferred embodiment of Formula II, B ² is

_R 16

wherein each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In one aspect of this embodiment, R ^3b is CH2R ⁹ wherein R ⁹ is not H. In another aspect of this embodiment, R ^3b is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3b is alkynyl or substituted alkynyl. In another aspect of this

15 embodiment, each E ² , E, and D is >N. In another aspect of this embodiment, each E ² , E, and D is >N and F ^x is >C-R ²⁵ . In another aspect of this embodiment, each E ² , E, and D is >N and each E ¹ and F" is independently XZ-R ²⁵ . In another aspect of this embodiment, each E ¹ , E ² , E, and D is >N and F ^x is >C-R ²⁵ . In another aspect of this embodiment, each E ¹ , E, and D is >N and each E ² and F* is

20 independently >C-R ²⁵ . In another aspect of this embodiment, each E and D is >N; each E ¹ and F* is independently >C-R ²⁵ ; and E ² is >C-R ³⁰ .

In another preferred embodiment of Formula II, B ² is

41

>16

N

'rv/

^V ^{^} wherein each E ² , E, and D is >N, each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In one aspect of this embodiment, R ^3b is CPi-R ⁹ wherein R ⁹ is not H. In another aspect of this embodiment, R ^3b is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3b is alkynyl or substituted alkynyl. In another aspect of this invention, F ^x is XZ-R ²⁵ . In another aspect of this invention

10 E ¹ is >N. In another aspect of this invention, E ¹ is XZ-R ²⁵ .

In another preferred embodiment of Formula II, B ² is

_R 16

wherein each E ² , E, and D is >N, F* is XI-R ²⁵ , each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In one aspect of this embodiment, R ^3b is CH2R ⁹ wherein R ⁹ is not H. In another aspect of this embodiment, R ^3b is alkenyl or substituted

15 alkenyl. In another aspect of this embodiment, R ^3b is alkynyl or substituted alkynyl. In another aspect of this embodiment, E ¹ is >N. In another aspect of this embodiment, E ¹ is XZ-R ²⁵ .

In another preferred embodiment of Formula II, B ² is

_R 16

wherein each E ² , E, and D is >N, each E ¹ and F ^x is independently

20 XI-R ²⁵ , each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In one aspect of this embodiment, R ^3b is CH2R ⁹ wherein R ⁹ is not H. In another aspect of this embodiment, R ^3b is

42

5 alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3b is alkynyl or substituted alkynyl. In another aspect of this embodiment, each R ²⁵ is H. In another aspect of this embodiment, R ¹⁵ is N(R ²⁰ )(R ²¹ ). In another aspect of this embodiment, R ¹⁶ is N(R ²⁰ )(R ²¹ ) wherein R ²⁰ is OR ^17a . In another aspect of this embodiment, R ¹⁶ is OR ^17a . In another aspect of this embodiment, R ¹⁶ is NH2 and

10 each R ²⁵ is H. In another aspect of this embodiment, R ¹⁶ is OR ^17a and F* is >C- NR ²⁶ R ²⁷ . In another aspect of this embodiment, R ¹⁶ Js OH, E ¹ is >C-R ²⁵ wherein R ²⁵ is H, and F ^x is >C-NR ²⁶ R ²⁷ wherein each R ²⁶ and R ²⁷ is H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R ^x is not H. In another aspect of this

15 embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is -O- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R ^x is not H.

In another preferred embodiment of Formula II, B ² is

_R 16

wherein each E ¹ , E ² , E, and D is >N, F ^x is >C-R ²⁵ , each R ¹⁰ is H, L ¹ 20 is O, and each R ^a is H. In one aspect of this embodiment, R ^3b is CH2R ⁹ wherein R ⁹ is not H. In another aspect of this embodiment, R ^3b is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3b is alkynyl or substituted alkynyl. In another aspect of this embodiment, R ¹⁶ is N(R ²⁰ )(R ²¹ ). In another aspect of this embodiment, R ¹⁶ is N(R ²⁰ )(R ²¹ ) wherein R ²⁰ is OR ^17a . In another 25 aspect of this embodiment, R ¹⁶ is OR ^17a . In another aspect of this embodiment, R ¹⁶ is NH2 and each R ²⁵ is H. In another aspect of this embodiment, R ¹⁶ is OR ^17a and F ^x is >C-NR ²⁶ R ²⁷ . In another aspect of this embodiment, R ¹⁶ is NH2 and each R ²⁵ is H. In another aspect of this embodiment, R ¹⁶ is OR ^17a and F ^x is >C-NR ²⁶ R ²⁷ .

43

5 In another aspect of this embodiment, R ¹⁶ is OH and F ^x is >C-NR ²⁶ R ²⁷ wherein each R ²⁶ and R ²⁷ is H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R* wherein each Y ² is -O- and R* is not H. In another aspect of this 10 embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R ^x is not H.

In another preferred embodiment of Formula II, B ² is

_R 16

^J ^ wherein each E ¹ , E, and D is >N, each E ² and F ^x is independently

XH-R ²⁵ , each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In one aspect of this embodiment, 15 R ^3b is CH2R ⁹ wherein R ⁹ is not H. In another aspect of this embodiment, R ^3b is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3b is alkynyl or substituted alkynyl. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is 20 independently Y ² -R ^x wherein each Y ² is -O- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R ^x is not H.

In another preferred embodiment of Formula II, B ² is

_R 16

F ^>

^J ^ wherein each E and D is >N, each E ¹ and F* is independently >C-

44

5 R ²⁵ , E ² is >C-R ³⁰ , each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In one aspect of this embodiment, R ^3b is CH2R ⁹ wherein R ⁹ is not H. In another aspect of this embodiment, R ^3b is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3b is alkynyl or substituted alkynyl. In another aspect of this embodiment R ¹⁶ is N(R ²⁰ )(R ²¹ ). In another aspect of this embodiment, R ¹⁶ is

10 N(R ²⁰ )(R ²¹ ) wherein R ²⁰ is OR ^17a . In another aspect of this embodiment, R ¹⁶ is OR ^17a . In another aspect of this embodiment, R ¹⁵ is NH2 and each R ²⁵ is H. In another aspect of this embodiment, R ¹⁶ is OR ^17a and F* is >C-NR ²⁶ R ²⁷ . In another aspect of this embodiment, R ¹⁶ is NH2 and each R ²⁵ is H. In another aspect of this embodiment, R ¹⁶ is OR ^17a and F ^x is >C-NR ²⁶ R ²⁷ . In another aspect of this

15 embodiment, R ¹⁶ is OH and F* is >C-NR ²⁶ R ²⁷ wherein each R ²⁶ and R ²⁷ is H. In another aspect of this embodiment, R ³⁰ is ethynyl, 2-trimethylsilylethynyl, 2-(2- pyridyl)ethynyl, 2-(4~pyridyl)ethynyl, 2-(4-methoxy)ethynyl, 2-(aminocarbonyl)ethynyl, 3,3-diethoxypropyn- 1-yl, 2-(dimethylaminocarbonyl)ethynyl, 2-(N-amino(aminocarbonyl)ethynyl,

20 2-carboxyethynyl, 2-ethoxycarbonylethynyl, 2-methoxycarbonylethynyl,

2-phenylethynyl, 2-(4-fluorophenyl)ethynyl, 2-(4-methylphenyl)ethynyl, vinyl, 2-methoxyvinyl, formyl, -CH=N-NFl-, -CH=NOH, 1,1-diisopropoxymethyl, or -B(OH) 2. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R* wherein each Y ² is independently -O- or -N(R)- and R ^x is

25 not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is -O- and R" is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R ^x is not H.

In another preferred embodiment of Formula II, B ² is

45

_R 45 41

R 44 _R

N N

- ^{O CX "} W 'R ⁴⁰

₃ 42

42

R ⁴³ -Y

R ⁴²

R ⁴³ -Y

R ⁴³ -Y

R ⁴³ - Y,

R ⁴³ -Y

,47 or . In one aspect of this embodiment, each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ^3b is CH2R ⁹ wherein R ⁹ is not H. In another aspect of this embodiment, R ^3b is alkenyl or substituted alkenyl. In another aspect of this

10 embodiment, R ^3b is alkynyl or substituted alkynyl. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is

46

independently -O- or -N(R)- and R* is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is -O- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R" is not H.

In another preferred embodiment of Formula II, B ² is

_R 45

R ^A t ,R ⁴¹

N' ^V N ^'

/

R ⁴³ - W Uλ ^• N / Y.

R ⁴²

R ⁴³ -γ

R ⁴³ -Y

10 j42

_R 43_γ

R ⁴³ -Y

N

R ⁴³ -Y _R 43_γ

_R 47 or wherein each R ¹⁰ is

47

H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ^3b is CHzR ⁹ wherein R ⁹ is not H. In another aspect of this embodiment R ^3b is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3b is alkynyl or substituted alkynyl. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R ^x is

10 not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is -O- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R ^x is not H.

In another preferred embodiment of Formula II, B ² is

N i

-R ⁵¹

_R 55

N N

_R 56

N

"Y^ or

15 ^" *- . In one aspect of this embodiment, each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ^3b is CH2R ⁹ wherein R ⁹ is not H. In another aspect of this embodiment, R ^3b is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3b is alkynyl or substituted alkynyl.

48

In another preferred embodiment of Formula II, B ² is

_R 50 _R 50

N

N

N- -R ⁵¹

_R 55

R ⁵ £

^N

N,

N R ⁵⁶ "N' ^* O

λλλ

*^v^ or e, ,NH wherein each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ^3b is CH2R ⁹ wherein R ⁹ is not H. In another aspect of this embodiment, R ^3b is alkenyl or substituted alkenyl. In another aspect of this embodiment R ^3b is

10 alkynyl or substituted alkynyl. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R* wherein each Y ² is independently -O- or -N(R)- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is -O- and R* is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² ~R* wherein each Y ² is

15 independently -N(R)- and R ^x is not H.

In another preferred embodiment of Formula ϊI, B ² is

49

vλλλ

' . In one aspect of this embodiment, each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ^3b is CH2R ⁹ wherein R ⁹ is not H. In another aspect of this embodiment, R ^3b is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3b is alkynyl or substituted alkynyl.

10 In another preferred embodiment of Formula II, B ² is

_R 60

_R 62 - _N - ^r ₀

wherein each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ^3b is CH2R ⁹ wherein R ⁹ is not H. In another aspect of this embodiment, R ^3b is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3b is alkynyl or substituted alkynyl. In another

15 aspect of this embodiment, R ⁶⁰ is NR ⁶¹³ R ⁶⁰ . In another aspect of this embodiment, R ⁶⁰ is NR ⁶¹³ R*= and R ⁶² is H. In another aspect of this embodiment, R ⁶⁰ is OR ^6b . In another aspect of this embodiment, R ⁶⁰ is OR ^6b and R ⁶² is H. In another aspect of this embodiment, each R ⁶¹ and R ⁶² is H and R ⁶⁰ is NH2. In another aspect of this embodiment, each R ⁶¹ and R ⁶² is H and R ⁶⁰ is OH. In another aspect of this

20 embodiment, R ⁶⁰ is not NR ⁶¹³ R ⁶⁰ or OR ^6b . In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is

50

5 independently Y ² -R ^x wherein each Y ² is -O- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R ^x is not H.

In another preferred embodiment of Formula II, B ² is

R70 _χ β ^R

^/υ N

. In one aspect of this embodiment, each R ¹⁰ is H, L ¹ is O, and 10 each R ^a is H. In another aspect of this embodiment, R ^3b is CH2R ⁹ wherein R ⁹ is not H. In another aspect of this embodiment, R ^3b is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3b is alkynyl or substituted alkynyl.

In another preferred embodiment of Formula II, B ² is

r ^x ,R ⁷⁰

N

15 ' ^" wherein each R ¹⁰ is H, L ¹ is O, and each R" is H. In another aspect of this embodiment, R ^3b is CH2R ⁹ wherein R ⁹ is not H. In another aspect of this embodiment, R ^3b is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3b is alkynyl or substituted alkynyl. In another aspect of this embodiment, X ^s is CH and each X ⁶ and X ⁷ is N. In another aspect of this

20 embodiment, X ⁵ is CH, each X ⁶ and X ⁷ is N, and R ⁷⁰ is -C(O)NR ⁷¹³ R^. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is -O- and

51

5 R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R ^x is not H.

In another preferred embodiment of Formula II, B ² is

X ⁶ . .R ⁷⁰

X ⁿ N , wherein X ⁵ is CH, each X ⁶ and X ⁷ is N, R ⁷⁰ is -C(O)NR ⁷¹⁵ R ⁷ =, each R ¹⁰ is H, L ¹ is O ₇ and each R ^a is H. In another aspect of this embodiment, R ^3b is

10 CH2R ⁹ wherein R ⁹ is not H. In another aspect of this embodiment, R ^3b is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3b is alkynyl or substituted alkynyl. In another aspect of this embodiment, each R ⁵ * and R ^7c is H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R" is not H. In another

15 aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is -O- and R* is not H. In another aspect of this embodiment, each W and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R ^x is not H.

In another preferred embodiment of Formula II, B ² is

52

γ80

^• N V ⁸⁰

A ⁸⁰ (( /

^^..«A* ₃ 78O

N' 'V ⁸⁰

. In one aspect of this embodiment, each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ^3b is CH2R ⁹ wherein R ⁹ is not H. In another aspect of this embodiment, R ^3b is alkenyl or substituted atkenyl. In another aspect of this embodiment, R ^3b is alkynyl or substituted alkynyl.

10 In another preferred embodiment of Formula II, B ² is

53

wherein each R ^1Q is

H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ^3b is CH2R ⁹ wherein R ⁹ is not H. In another aspect of this embodiment R ^3b is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3b is alkynyl or substituted alkynyl. In another aspect of this embodiment, each W ¹ and W ² is 10 independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is -O- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y^R" wherein each Y ² is independently -N(R)- and R ^x is not H.

15 In another preferred embodiment of Formula II, B ² is

54

. In one aspect of this embodiment, each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ^3b is CH2R ⁹ wherein R ⁹ is not H. In another aspect of this embodiment, R ^3b is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3b is alkynyl or substituted alkynyl.

10 In another preferred embodiment of Formula II, B ² is R ⁹⁰

wherein each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ^3b is CH2R ⁹ wherein R ⁹ is not H. In another aspect of this embodiment, R ^3b is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3b is alkynyl or substituted alkynyl. In another aspect of this

15 embodiment, X ⁹ is CR ^9Oa . In another aspect of this embodiment, X ¹⁰ is O, S, or NR ^91a . In another aspect of this embodiment, X ⁹ is CH and X ¹⁰ is O, S, or NR ^91a . In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each

20 Y ² is -O- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R" is not H.

In another preferred embodiment of Formula II, B ² is

55

wherein X ⁹ is CH, X ¹⁰ is O, S, or NR ⁹¹³ , each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ^3b is CH2R ⁹ wherein R ⁹ is not H. In another aspect of this embodiment, R ^3b is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3b is alkynyl or substituted alkynyl. In another aspect of this embodiment, X ¹⁰ is O. In another aspect of this

10 embodiment, X ¹⁰ is S. In another aspect of this embodiment, X ¹⁰ is NR ^91a . In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² ^-R* wherein each Y ² is -O- and R* is not H. In another aspect of this embodiment, each W ¹ and W ²

15 is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R ^x is not H.

In another preferred embodiment of Formula II, B ² is

V

«λ/Xγ> . In one aspect of this embodiment, each R ¹⁰ is H, JJ is O, and each R ^a is H. In another aspect of this embodiment, R ^3b is CH2R ⁹ wherein R ⁹ is not H. In another aspect of this embodiment, R ^3b is alkenyl or substituted alkenyl. In

20 another aspect of this embodiment, R ^3b is alkynyl or substituted alkynyl. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R ^x is not H. In another aspect of this embodiment each W ¹ and W ² is independently Y ² -R ^x wherein each

56

Y ² is -O- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R* wherein each Y ² is independently -N(R)- and R ^x is not H.

In another preferred embodiment of Formula II, B ² is

Z ¹ Z\ Z ¹ rZ{

/ \r > \λ/6 *' \c \ \λ/6

_W 6 / ^W \ W ^β ^ ^W \

// vy ⁶ ..--- ^* *, N /'' VV ⁶ , _,' - ^" ON ² I' ^' I ^•' ; I ^z ϊ> 1 i '

P ^z ^1' r ""z ¹

' or ' . In one aspect of this embodiment, each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this 10 embodiment, R ^3b is CH2R ⁹ wherein R ⁹ is not H. In another aspect of this embodiment, R ^3b is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3b is alkynyl or substituted alkynyl.

In another preferred embodiment of Formula II, B ² is z ¹ -

\

X ⁸

Z ¹ '. x ^β / .' - ² \ z ¹ •:

W ⁶

% N I /' ^{' V} N

I ; I I

/ I Z ¹ ' ^z \ Z ¹ .Wf--' ^' Z ¹

' ox I w ^{^} herein each R ^:o is H, L ¹ is O,

15 and each R ^a is H. In another aspect of this embodiment, R ^3b is CH2R ⁹ -wherein R ⁹ is not H. In another aspect of this embodiment, R ^3b is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3b is alkynyl or substituted alkynyl. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R ^x is not H. In 20 another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x

57

wherein each Y ² is -O- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R* is not H. In another aspect of this embodiment, a specific value for B ² is

P

58

PCH ₃

NH

\ N N ^' Cl 'N' ^* O ¹ A-

T^

CH ₃

T ⁱ NH

'N' O

α. ^

H ₃ CO- or

⁴ A-

In another preferred embodiment of Formula II, B ² is

59

N NH ₃

<

60

.OCH

N' J** wherein each R ¹⁰ is H, L ¹ is O, and each. R ^a is H. In another aspect of this embodiment, R ^3b is CH2R ⁹ wherein R ⁹ is not H. In another aspect of this embodiment, R ^3b is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3b is alkynyl or substituted

10 alkynyl. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is -O- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² ~R ^X wherein each Y ² is independently -N(R)-

15 and R ^x is not H.

In one embodiment, compounds of this invention are of Formula III: ^•

61

O wi w * ^{: *} I B ²

R 3c R ⁸

R ¹ 6- -R 7

5 R ⁵ OR ^a

Formula III wherein all variables are defined as above for Formula III. In one aspect of this embodiment, R ^a is H. In another aspect of this embodiment, each R ¹⁰ is H and L ¹ is O. In another aspect of this embodiment R ^3c is CH2R ⁹ wherein R ⁹ is not H, OH, 10 or F. In another aspect of this embodiment, R ^3c is alkenyl or substituted alkenyl. In another aspect of this embodiment R ³ * is alkynyl or substituted alkynyl. In another aspect of this embodiment each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ⁵ and R ⁶ , taken together, are =CR ^c R ^d .

In a preferred embodiment of Formula IE, B ² is

_R 1 ^β

15 ^i . In one aspect of this embodiment, each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ^3c is CH2R ⁹ wherein R ⁹ is not H, OH, or F. In another aspect of this embodiment, R ³⁰ is alkenyl or substituted alkenyl. In another aspect of this embodiment, R^ is alkynyl or substituted alkynyl. In another aspect of this embodiment, each R ⁵ and R ⁶ is H.

20 In another aspect of this embodiment, R ⁵ and R ⁶ , taken together, are =CR ^c R ^d .

In another preferred embodiment of Formula III, B ² is

62

_R 16

E ² -^/^

5 C ^J L wherein each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ³ *= is CH2R ⁹ wherein R ⁹ is not H, OH, or F. In another aspect of this embodiment, R ^3c is alkenyl or substituted, alkenyl. In another aspect of this embodiment, R ³⁰ is alkynyl or substituted alkynyl. In another aspect of this embodiment, each R ⁵ and R ⁶ is H. In another aspect of this

10 embodiment, R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, each E ² , E, and D is >N. In another aspect of this embodiment, each E ² , E, and D is >N and F ^x is >C-R ²⁵ . In another aspect of this embodiment, each E ² , E, and D is >N and each E ¹ and F* is independently >C-R ²⁵ . In another aspect of this embodiment, each E ¹ , E ² , E, and D is >N and F is ^-R ²⁵ . In another

15 aspect of this embodiment, each E ¹ , E, and D is >N and each E ² and F ^x is independently >C-R ²⁵ . In another aspect of this embodiment, each E and D is >N; each E ¹ and F" is independently XZ-R ²⁵ ; and E ² is >C-R ³⁰ .

In another preferred embodiment of Formula III, B ² is

_R 1β

wherein each E ² , E, and D is >N, each R ¹⁰ is H, L ¹ is O, and each 20 R ^a .is H. In another aspect of this embodiment, R ³⁰ is CH2R ⁹ wherein R ⁹ is not H, OH, or F. In another aspect of this embodiment, R ³⁰ is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3c is alkynyl or substituted alkynyl. In another aspect of this embodiment, each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another 25 aspect of this invention, F ^x is >C-R ²⁵ . In another aspect of this invention E ¹ is >N.

63

5 In another aspect of this invention, E ¹ is >C-R ²⁵ .

In another preferred embodiment of Formula III, B ² is

_R 16

wherein each E ² , E, and D is >N, F ^x is >C-R ²⁵ , each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ³⁰ is CH2R ⁹ wherein R ⁹ is not H, OH, or F. In another aspect of this embodiment, R ³¹ = is alkenyl or 10 substituted alkenyl. In another aspect of this embodiment, R ³ = is alkynyl or substituted alkynyl. In another aspect of this embodiment, each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, E ¹ is >N. In another aspect of this embodiment, E ^: is X-Z-R ²⁵ .

15 In another preferred embodiment of Formula III, B ² is

_R 16

wherein each E ² , E, and D is >N, each E ¹ and F ^x is independently XI-R ²⁵ , each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ³⁰ is CH2R ⁹ wherein R ⁹ is not H, OH, or F. In another aspect of this embodiment, R ³¹ is alkenyl or substituted alkenyl. In another aspect of this 20 embodiment, R ³⁰ is alkynyl or substituted alkynyl. In another aspect of this embodiment, each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, each R ²⁵ is H. In another aspect of this embodiment, R ¹⁶ is N (R ²⁰ ) (R ²¹ ). In another aspect of this embodiment, R ¹⁶ is N(R ^2O )(R ²¹ ) wherein R ²⁰ is OR ¹⁷ⁿ . In another aspect of this

64

5 embodiment, R ¹⁶ is OR ^]7a . In another aspect of this embodiment, R ¹⁶ is NH2 and each R ²⁵ is H. In another aspect of this embodiment, R ¹⁶ is OR ^α7a and F" is >C- NR ²⁶ R ²⁷ . In another aspect of this embodiment, R ¹⁶ is OH, E ¹ is >C-R ⁷⁵ wherein R ²⁵ is H, and F ^x is >C-NR ²⁶ R ²⁷ wherein each R ²⁶ and R ²⁷ is H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is 10 independently -O- or -N(R)- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is -O- and R* is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R ^x is not H.

In another preferred embodiment of Formula III, B ² is

_R 16

15 ^V wherein each E ¹ , E ² , E, and D is >N, F* is XZ-R ²⁵ , each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ³⁰ is CH2R ⁹ wherein R ⁹ is not H, OH, or F. In another aspect of this embodiment, R ^3c is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3c is alkynyl or substituted alkynyl. In another aspect of this embodiment, each R ⁵

20 and R ⁶ is H. In another aspect of this embodiment, R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, R ¹⁶ is N(R ²⁰ )(R ²¹ ). In another aspect of this embodiment, R ¹⁶ is N(R ²⁰ )(R ²¹ ) wherein R ²⁰ is OR ^17a . In another aspect of this embodiment, R ¹⁶ is OR ^17a . In another aspect of this embodiment, R ¹⁶ is NH2 and each R ²⁵ is H. In another aspect of this embodiment, R ¹⁶ is OR ^17a

25 and F ^x is >C-NR ²⁶ R ²⁷ . In another aspect of this embodiment, R ¹⁶ is NH2 and each R ²⁵ is H. In another aspect of this embodiment, R ¹⁶ is OR ^17a and F ^x is >C-NR ²⁶ R ²⁷ . In another aspect of this embodiment, R ¹⁶ is OH and F ^x is >C-NR ²⁶ R ²⁷ wherein each R ²⁶ and R ²⁷ is H. In another aspect of this embodiment, each W ¹ and W ² is

65

5 independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² τR ^x wherein each Y ² is -O- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein, each Y ² is independently -N(R)- and R ^x is not H.

10 . In another preferred embodiment of Formula III, B ² is

_R 16

wherein each E ¹ , E, and D is >N, each E ² and F ^x is independently >C-R ²⁵ , each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ³ = is CH2R ⁹ wherein R ⁹ is not H, OH, or F. In another aspect of this embodiment, R ^3c is alkenyl or substituted alkenyl. In another aspect of this

15 embodiment, R ^3c is alkynyl or substituted alkynyl. In another aspect of this embodiment, each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is

20 independently Y ² -R ^x wherein each Y ² is -O- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R ^x is not H.

In another preferred embodiment of Formula III, B ² is

_R 16

"^ wherein each E and D is >N, each E ¹ and F ^x is independently >C-

66

5 R ²⁵ , E ² is XZ-R ³⁰ , each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ³⁰ is CH2R ⁹ wherein R ⁹ is not H, OH, or F. In another aspect of this embodiment, R ^3c is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3c is alkynyl or substituted alkynyl. In another aspect of this embodiment, each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ⁵ and

10 R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, R ¹⁶ is

N(R ²⁰ )(R ²¹ ). In another aspect of this embodiment, R ¹⁶ is N(R ²⁰ )(R ²¹ ) wherein R ²⁰ is OR ^17a . In another aspect of this embodiment, R ¹⁶ is OR ^:7a . In another aspect of this embodiment, R ¹⁶ is NH2 and each R ²⁵ is H. In another aspect of this embodiment, R ¹⁶ is OR ^17a and F* is >C-NR ²⁶ R ²⁷ . In another aspect of this

15 embodiment, R ¹⁶ is NH2 and each R ²⁵ is H. In another aspect of this embodiment, R ¹⁶ is OR ^17a and F ^x is >C-NR ²⁶ R ²⁷ . In another aspect of this embodiment, R ¹⁶ is OH and F ^x is >C-NR ²⁶ R ²⁷ wherein each R ²⁶ and R ²⁷ is H. In another aspect of this embodiment, R ³⁰ is ethynyl, 2-trimethylsilylethynyl, 2-(2-pyridyl)ethynyl, 2-(4- pyridyl)ethynyl, 2-(4-methoxy)ethynyl, 2-(aminocarbonyl)ethynyl, 3,3-

20 diethoxypropyn-1-yl, 2-(dimethylaminocarbonyl)ethynyl, 2-(N- amino(aminocarbonyl)ethynyl, 2-carboxyethynyl, 2-ethoxycarbonylethynyl, 2- methoxycarbonylethynyl, 2-phenylethynyl, 2-(4-fluorophenyl)ethynyl, 2-(4- -methylphenyl) ethynyl, vinyl, 2-methoxyvinyl, formyl, -CH=N-NHz, -CH=NOH, 1,1-diisopropoxymethyl, or -B(OH) 2. In another aspect of this embodiment; each

25 W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or - ■ N(R)- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is -O- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)r and R ^x is not H.

30 In another preferred embodiment of Formula III, B ² is

67

R ⁴³ -Y.

₃ 42

_R 44

,42

R ⁴³ ~Y

R ⁴³ -Y

R ⁴³ ~Y

R ⁴³ -Y

R ⁴³ -Y.

R ⁴³ — Y or . In one aspect of this embodiment, each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ^3c is CH2R ⁹ wherein R ⁹ is not H, OH, or F. In another aspect of this embodiment, R ^3c is alkenyl or substituted alkenyl. In another aspect of

10 this embodiment, R ^3c is alkynyl or substituted alkynyl. In another aspect of this embodiment, each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ⁵ and

68

R ⁶ , taken together, are =CR ^c R ^d .

In another preferred embodiment of Formula III, B ² is

_R 45

R ⁴³ -Y

R ⁴³ -Y

R ⁴³ -Y

R ⁴³ -Y ?43_ Y

N

R ⁴³ - Y

R ⁴³ -Y

X or ^* W "R ⁴⁷ vrv/vr wherein each R ¹⁰ is

H, L ¹ is O, and each R ⁰ is H. In another aspect of this embodiment, R ^3c is CH2R ⁹

10 wherein R ⁹ is not H, OH, or F. In another aspect of this embodiment, R ^{3c :} iis; alkenyl or substituted alkenyl. In another aspect of this embodiment, R ³ = ^: iifs

69

alkynyl or substituted alkynyl. In another aspect of this embodiment, each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x

10 wherein each Y ² is -O- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R* wherein each Y ² is independently -N(R)- and R ^x is not H.

In another preferred embodiment of Formula III, B ² is

N — R ⁵¹

_R 55 R ⁵⁵

R ⁵ D57 M R58

V4 ^

N.

N

«λ/Vι or

I X ^H . In one aspect of this

15 embodiment, each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ³ = is CH2R ⁹ wherein R ⁹ is not H, OH, or F. In another aspect of this embodiment, R ^3c is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ³⁰ is alkynyl or substituted alkynyl. In another aspect of this embodiment, each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ⁵ and

20 R ⁶ , taken together, are =CR ^c R ^d .

In another preferred embodiment of Formula III, B ² is

70

_R 55 _R 55

N, .R ^{58 Y R} ^ ^N Y Y

N N T NH wherein R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ³⁰ is CH2R ⁹ wherein R ⁹ is not H, OH, or F. In another aspect of this embodiment, R ^3c is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ³⁰ is alkynyl or substituted alkynyl. In another aspect of this embodiment, each R ⁵

10 and R ⁶ is H. In another aspect of this embodiment, R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y^R* wherein each Y ² is -O- and R ^x is not H. In another aspect of this embodiment,

15 each W ^: and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R* is not H.

In another preferred embodiment of Formula III, B ² is

_R 60

N

_R62 ^ _N A, _O

In one aspect of this embodiment,, each R ¹⁰ is H, L ¹ is O, and

71

5 each R ^a is H. In another aspect of this embodiment, R ³ = is CH2R ⁹ wherein R ⁹ is not H, OH, or F. In another aspect of this embodiment, R ^3c is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3c is alkynyl or substituted alkynyl. In another aspect of this embodiment, each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ⁵ and R ⁶ ₇ taken together, are =CR ^c R ^d .

10 In another preferred embodiment of Formula III, B ² is

_R 60

1

_R 61

wherein each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment _/ R ³⁰ is CH2R ⁹ wherein R ⁹ is not H, OH, or F. In another aspect of this embodiment, R ^3c is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3c is alkynyl or substituted alkynyl. In

15 another aspect of this embodiment, each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, R ⁶⁰ is NR ^6b R ^6c . In another aspect of this embodiment, R ⁶⁰ is NR ^6b R ^6c and R ⁶² is H. In another aspect of this embodiment, R ⁶⁰ is OR ^6b . In another aspect of this embodiment/ R ⁶⁰ is OR ^6b and R ⁶² is H. In another aspect of this

20 embodiment, R ⁶⁰ is OH and each R ⁶¹ and R ⁶² is H. In another aspect of this embodiment, R ⁶⁰ is NH2 and each R ⁶¹ and R ⁶² is H. In another aspect of this embodiment, R ⁶⁰ is not NR ^6b R ^6c or OR ^6b . In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is

25 independently Y ² -R ^x wherein each Y ² is -O- and R* is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R ^x is not H.

In another preferred embodiment of Formula III, B ² is

72

_χ6 ^R ^{70 f~X} .

5 / ^v v j _{n one} aspect _o f ^ ₃ embodiment, each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ³⁰ is CH2R ⁹ wherein R ⁹ is not H, OH, or F. In another aspect of this embodiment, R^ is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3c is alkynyl or substituted alkynyl. In another aspect of this embodiment, each R ⁵ and R ⁶ is H. 10 In another aspect of this embodiment, R ⁵ and R ⁶ , taken together, are =CR ^c R ^d .

In another preferred embodiment of Formula III, B ² is

R70

_X 6 _^ R

^/T N

wherein each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ^3c is CH2R ⁹ wherein R ⁹ is not H, OH, or F. In another aspect of this embodiment, R ³ *= is alkenyl or substituted alkenyl. In

15 another aspect of this embodiment, R ^3c is alkynyl or substituted alkynyl. In another aspect of this embodiment, each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, X ⁵ is CH and each X ⁶ and X ⁷ is N. In another aspect of this embodiment, X ⁵ is CH, each X ⁶ and X ⁷ is N, and R ⁷⁰ is -C(O)NR ⁷¹⁵ R ⁷⁰ . In another

20 aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R* is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is -O- and R* is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R ^x is not H.

73

In another preferred embodiment of Formula III, B ² is

_R 70

N

wherein X ⁵ is CH, each X ⁶ and X ⁷ is N, R ⁷⁰ is -C(O)NR ⁷ Wv each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ^3c is CH2R ⁹ wherein R ⁹ is not H, OH, or F. In another aspect of this embodiment, R ^3c is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ³⁰ is

10 alkynyl or substituted alkynyl. In another aspect of this embodiment, each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x

15 wherein each Y ² is -O- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R" wherein each Y ² is independently -N(R)- and R* is not H.

In another preferred embodiment of Formula III, B ² is

74

ϊS- _V 8O

^■ N'

A. -80

vλλλ o ^f / . In one aspect of this embodiment each R ¹⁰ is H ₇ L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ^3c is CH2R ⁹ wherein R ⁹ is not H, OH, or F. In another aspect of this embodiment, R ^3c is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ³ = is alkynyl or substituted alkynyl. In another aspect of this

10 embodiment, each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is -O- and R ^x is not H. In another aspect of

15 this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R ^x is not H.

In another preferred embodiment of Formula III, B ² is

75

. In one aspect of this embodiment, each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ³ = is CHzR ⁹ wherein R ⁹ is not H, OH, or F. In another aspect of this embodiment, R ³⁰ is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ³ *= is alkynyl or substituted alkynyl. In another aspect of this embodiment, each R ⁵ and R ⁵ is H.

10 In another aspect of this embodiment, R ⁵ and R ⁶ , taken together, are =CR ^c R ^d .

In another preferred embodiment of Formula III, B ² is R ⁹⁰

X ¹⁰ .

wherein each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ^3c is CH2R ⁹ wherein R ⁹ is not H, OH, or F. In another aspect of this embodiment, R ^3c is alkenyl or substituted alkenyl. In

15 another aspect of this embodiment, R ^3c is alkynyl or substituted alkynyl. In another aspect of this embodiment, each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, X ⁹ is CR ^90a . In another aspect of this embodiment; X ¹⁰ is O, S, or NR ^91a . In another aspect of this embodiment, X ⁹ is CH and X ¹⁰ is O, S, or NR ^91a .

20 In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R* is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is -O- and R* is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R* wherein each Y ² is independently -N(R)- and R* is not H.

76

In another preferred embodiment of Formula III, B ² is

_R 90

wherein X ⁹ is CH, X ¹⁰ is O, S, or NR ^91a , each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ^3b is CH2R ⁹ wherein R ⁹ is not H. In another aspect of this embodiment, R ³ ^ is CH2R ⁹ wherein R ⁹ is not H, OH, or F. In another aspect of this embodiment, R ^3c is alkenyl or substituted

10 alkenyl. In another aspect of this embodiment, R ³⁰ is alkynyl or substituted alkynyl. In another aspect of this embodiment, each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, X ¹⁰ is O. In another aspect of this embodiment, X ¹⁰ is S. In another aspect of this embodiment, X ¹⁰ is NR ^91a . In another aspect of this

15 embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R" is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is -O- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R* is not H.

20 In another preferred embodiment of Formula III, B ² is

/ \

^I/W . In one aspect of this embodiment, each R ¹⁰ is H, L ¹ is O, and each

R ^a is H. In another aspect of this embodiment, R ^3c is CH2R ⁹ wherein R ⁹ is not H,

77

5 OH, or F. In another aspect of this embodiment, R ^3c is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ³ = is alkynyi or substituted alkynyl. In another aspect of this embodiment, each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each 10 Y ² is independently -O- or -N(R)- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is -O- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R ^x is not H.

In another preferred embodiment of Formula III, B ² is z ¹ - z ¹ -

X ⁸ - \. ^z v ^"

/ \ ^ /S I ' » 1

_Z 1 // I - I I φ Z? I ^■ \ ' MI

V N

"Z ¹ '* V ^wS ^* -- .z

^Z ^

15 I' oorr / / . In one aspect of this embodiment, each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ³⁰ is CH2R ⁹ wherein R ⁹ is not H, OH, or F. In another aspect of this embodiment, R ^3c is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3c is alkynyl or substituted alkynyl. In another aspect of this

20 embodiment, each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ⁵ and R ⁶ , taken together, are =CR ^c R ^d .

In another preferred embodiment of Formula III, B ² is

78

Z ¹ - Z ¹ Z ¹ / .' z ^{1 ■} : X ⁸

W ⁶ . .Wl

N /' * ^{■ "} W ^6' 'N

Z ¹ -W^- - ' ^' ^Z ¹

¹ ^ >z ¹ ' or wherein each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R^ is CH2R ⁹ wherein R ⁹ is not H, OH, or F. In another aspect of this embodiment, R^ is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ³⁰ is alkynyl or substituted alkynyl. In another aspect of this embodiment, each R ⁵ and R ⁵ is H.

10 In another aspect of this embodiment, R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R" is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R" wherein each Y ² is — O- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ²

15 is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R ^x is not H. In another aspect of this embodiment, a specific value for B ² is

P

NH

N

NH

/

\ 1 « j

%.

19

P ^Br v P

NH H ₃ CO- NH

H ₃ CO J

N

/ J J

, ^N N^ ^CI

In another preferred embodiment of Formula III, B ² is

80

NH

NH

Br o

H ₃ CO / X NH

%,

81

,OCH ₃

wherein each R ¹⁰ is H, L ¹ is O, and each R ^a is H. In another aspect of this embodiment, R ^3c is CH2R ⁹ wherein R ⁹ is not H, OH, or F. In another aspect of this embodiment, R ^3c is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ³ * ¹ is alkynyl or

10 substituted alkynyl. In another aspect of this embodiment, each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently — O- or -N(R)- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each

15 Y ² is -O- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R" is not H.

In one embodiment, compounds of this invention are of Formula IV:

82

Wi

W ² ' H. B ³

_R 3d v-λv r~-;R ^{β R6} I I ^R7

5 R ⁵ OR ^a

Formula IV wherein all variables are defined as above for Formula IV. In one aspect of this embodiment, R ^a is H. In another aspect of this embodiment, each R ¹⁰ is H and L ¹ is O. In another aspect of this embodiment, R ^M is H. In another aspect of this

10 embodiment, R^ is CH2R ⁹ . In another aspect of this embodiment, R ^3d is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ³ * ¹ is alkynyl or substituted alkynyl. In another aspect of this embodiment, each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, R ^a is H, each R ¹⁰ is H and L ¹ is O. In another

15 aspect of this embodiment, R ^a is H, each R ¹⁰ is H, L ¹ is O and R ^3d is H. In another aspect of this embodiment, R ^a is H, each R ¹⁰ is H, L ¹ is O and R ^3d is CH2R ⁹ . In another aspect of this embodiment, R ^a is H, each R ¹⁰ is H, L ¹ is O and R ^3d is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^a is H, each R ¹⁰ is H, L ¹ is O and R ^3d is alkynyl or substituted alkynyl.

20 In a preferred embodiment of Formula IV, B ³ is

• _R 1β

I ^' . In one aspect of this embodiment, R ^a is H. In another aspect of this embodiment, each R ¹⁰ is H and L ¹ is O. In another aspect of this embodiment, R™ is H. In another aspect of this embodiment, R ³ * is CBbR ⁹ . In another aspect of this embodiment, R ^3d is alkenyl or substituted alkenyl. In 25 another aspect of this embodiment, R ³ * ¹ is alkynyl or substituted alkynyl. In

83

5 another aspect of this embodiment, each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, R ^a is H, each R ¹⁰ is H and L ¹ is O. In another aspect of this embodiment, R ^a is H, each R ¹⁰ is H, L ¹ is O and R ^M is H. In another aspect of this embodiment, R ^a is H, each R ¹⁰ is H, L ¹ is O and R ^3d is CH2R ⁹ . In another aspect of 10 this embodiment, R ^a is H, each R ¹⁰ is H, L ¹ is O and R ^3d is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^a is H, each R ¹⁰ is H, L ¹ is O and R ^3d is alkynyl or substituted alkynyl. In another aspect of this embodiment, each E and D is >N. In another aspect of this embodiment, each E and D is >N and F ^x is XI-R ²⁵ .

15 In another preferred embodiment of Formula IV, B ³ is

_R 16

* ^r * ^λ ^ wherein R ^a is H, each R ¹⁰ is H and L ¹ is O. In another aspect of this embodiment, R ^3d is H. In another aspect of this embodiment, R ^3d is CH2R ⁹ . In another aspect of this embodiment, R ^3d is alkenyl or substituted alkenyl. In another aspect of this embodiment, R^ is alkynyl or substituted alkynyl. In 20 another aspect of this embodiment, each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ⁵ and R ⁶ , taken together, are ==CR ^c R ^d . In another aspect of this embodiment, each E and D is >N. In another aspect of this embodiment, each E and D is >N and F* is XI-R ²⁵ .

In another preferred embodiment of Formula IV, B ³ is

84

_R 16

N-^/^ ⁵

<X N ^r J- wherein R ^a is H, each R ¹⁰ is H, L ¹ is O and each E and D is >N. In another aspect of this embodiment, R ^3d is H. In another aspect of this embodiment, R ^3d is CH2R ⁹ . In another aspect of this embodiment, R ^3d is alkenyl . or substituted alkenyl. In another aspect of this embodiment, R ^3d is alkynyl or substituted alkynyl. In another aspect of this embodiment, each R ⁵ and R ⁶ is H.

10 In another aspect of this embodiment, R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, F ^x is >C-R ²⁵ . In another aspect of this embodiment, R ¹⁶ is N(R ²⁰ )(R ²¹ ). In another aspect of this embodiment, R ¹⁶ is OR ^17a . In another aspect of this embodiment, R ¹⁶ is N(R ²⁰ )(R ²¹ ) and F* is >C-H. In another aspect of this embodiment, R ¹⁶ is OR ^17a and F* is >C-NR ²⁶ R ²⁷ .

15 In another preferred embodiment of Formula IV, B ³ is

_R 16

wherein R ^a is H, each R ¹⁰ is H, L ¹ is O, each E and D is >N and F" is >C-R ²⁵ . In another aspect of this embodiment, R ^3d is H. In another aspect of this embodiment, R ^M is CH2R ⁹ . In another aspect of this embodiment, R ^3d is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3d is

20 alkynyl or substituted alkynyl. In another aspect of this embodiment, each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, R ¹⁶ is N(R ²⁰ )(R ²¹ )- In another aspect of this embodiment, R ¹⁶ is OR ^17a . In another aspect of this embodiment, R ¹⁶ is N(R ²⁰ XR ²¹ ) and R ²⁵ is H. In another aspect of this embodiment, R ¹⁶ is OR ^17a

25 and R ²⁵ is NR ²⁶ R ²⁷ . In another aspect of this embodiment, R ¹⁶ is OH and R ²⁵ is

85

5 NH2. In another aspect of this embodiment, R ¹⁶ is NHb and R ²⁵ is H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R* is not H.- In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is -O- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is 10 independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R* is not H.

In another preferred embodiment of Formula IV, B ³ is

_R 16

A . wherein R ^a is H, each R ¹⁰ is H, L ¹ is O, each E and D is >N, F ^x is

5"C-R ²⁵ and each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ^3d is H. In another aspect of this embodiment, R ^M is CH2R ⁹ . In another aspect of this

15 embodiment, R ^M is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^M is alkynyl or substituted alkynyl. In another aspect of this embodiment, R ¹⁶ is N(R ²⁰ )(R ²¹ ). In another aspect of this embodiment, R ¹⁶ is OR ^17a . In another aspect of this embodiment, R ¹⁶ is N(R ²⁰ ) (R ²¹ ) and R ²⁵ is H. In another aspect of this embodiment, R ¹⁶ is OR ^17a and R ²⁵ is NR ²⁶ R ²⁷ . In another aspect of

20 this embodiment, R ¹⁶ is OH and R ²⁵ is NH2. In another aspect of this embodiment, R ¹⁶ is NH2 and R ²⁵ is H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R* wherein each Y ² is independently -O- or - N(R)- and R* is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is -O- and R ^x is not H. In another aspect of

25 this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R ^x is not H.

In another preferred embodiment of Formula IV, B ³ is

86

R ¹⁶

5 - ^C ^ ^O wherein R ^a is H, each R ¹⁰ is H, L ¹ is O, each E and D is >N, F" is

>C-R ²⁵ and R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, W ⁰ is H. In another aspect of this embodiment, R ^3d is CH2R ⁹ . In . another aspect of this embodiment, R ^3d is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3d is alkynyl or substituted alkynyl. In

10 another aspect of this embodiment, R ¹⁶ is N(R ²⁰ )(R ²¹ ). In another aspect of this embodiment, R ¹⁶ is OR ^17a . In another aspect of this embodiment R ¹⁶ is N(R ²⁰ ) (R ²¹ ) and R ²⁵ is H. In another aspect of this embodiment, R ¹⁶ is OR ^:7a and R ²⁵ is NR ²⁶ R ²⁷ . In another aspect of this embodiment, R ¹⁶ is OH and R ²⁵ is NH2. In another aspect of this embodiment, R ¹⁶ is NH2 and R ²⁵ is H. In another aspect of this

15 embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R* is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is -O- and R* is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R ^x is not H.

20 In another preferred embodiment of Formula IV ₇ B ³ is

R16

E ² ,

/ ^f E

-N 1

^" * ^D . In one aspect of this embodiment, R ⁰ is H. In another aspect of this embodiment, each R ¹⁰ is H and L ¹ is O. In another aspect of this embodiment, R ^3d is H. In another aspect of this embodiment, R ^M is CH2R ⁹ . In another aspect of this embodiment, R ^3d is alkenyl or substituted alkenyl. In 25 another aspect of this embodiment, R ^3d is alkynyl or substituted alkynyl. In

87

5 another aspect of this embodiment, each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, R ^a is H, each R ¹⁰ is H and L ¹ is O. In another aspect of this embodiment, R ^a is H, each R ¹⁰ is H, L ¹ is O and R ^3d is H. In another aspect of this embodiment, R ^a is H, each R ¹⁰ is H, L ¹ is O and R ^3d is CH2R ⁹ . In another aspect of

10 this embodiment, R ^a is H, each R ¹⁰ is H, L ¹ is O and R ³ " ¹ is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^a is H, each R ¹⁰ is H, L ¹ is O and R ^3d is alkynyl or substituted alkynyl. In another aspect of this embodiment, each E and D is >N. In another aspect of this embodiment, each E and D is >N and F ^x is >C-R ²⁵ . In another aspect of this embodiment, E ² is >N. In another aspect of .

15 this embodiment, E ² is XI-R ²⁵ . In another aspect of this embodiment, E ² is >C-R ³⁰ .

In another preferred embodiment of Formula IV, B ³ is

R16

E

^D wherein R ^a is H, each R ¹⁰ is H and L ¹ is O. In another aspect of this embodiment, R ^3d is H. In another aspect of this embodiment, R ^3d is CH2R ⁹ . In another aspect of this embodiment, R ^3d is alkenyl or substituted alkenyl. In

20 another aspect of this embodiment, R ³ * ¹ is alkynyl or substituted alkynyl. In another aspect of this embodiment, each R ⁵ and R ⁵ is H. In another aspect of this embodiment, R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, R ^a is H, each R ¹⁰ is H and L ¹ is O. In another aspect of this embodiment, R ^a is H, each R ¹⁰ is H, L ¹ is O and R ^3d is H. In another aspect of this

25 embodiment, R ^a is H, each R ¹⁰ is H, L ¹ is O and R ^3d is CH2R ⁹ . In another aspect of this embodiment, R ^a is H, each R ¹⁰ is H, L ¹ is O and R ^3d is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^a is H, each R ¹⁰ is H, L ¹ is O and R ^3d is alkynyl or substituted alkynyl. In another aspect of this embodiment, each E and D is >N. In another aspect of this embodiment, each E and D is >N and F*

88

5 is >C-R ²⁵ . In another aspect of this embodiment, E ² is >N. In another aspect of this embodiment E ² is >C-R ²⁵ . In another aspect of this embodiment, E ² is >C-R ³⁰ . In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each 10 Y ² is -O- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R ^x is not H.

In another preferred embodiment of Formula IV, B ³ is p70 ^r X ⁶ - ^R

V NT ^e

I ^ ^" . In one aspect of this embodiment, R ^a is H, each R ¹⁰ is H and L ¹ is O. In another aspect of this embodiment, R ^3d is H. In another aspect of this

15 embodiment, R ^M is CH2R ⁹ . In another aspect of this embodiment, R ^3d is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^3d is alkynyl or substituted alkynyl. In another aspect of this embodiment, each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, R ^a is H, each R ¹⁰ is H, L ¹ is O and R ³⁰ is H. In

20 another aspect of this embodiment, R ^a is H, each R ¹⁰ is H, L ¹ is O and R ^M is CH2R ⁹ . In another aspect of this embodiment, R ^a is H, each R ¹⁰ is H, L ¹ is O and R ³⁰ is alkenyl or substituted alkenyl. In another aspect of this embodiment, R ^a is H, each R ¹⁰ is H, L ¹ is O and R ^3d is alkynyl or substituted alkynyl.

In another preferred embodiment of Formula IV, B ³ is

89

.R ⁷⁰

X ¹

5 I wherein R ^a is H, each R ¹⁰ is H and L ¹ is O. In another aspect of this embodiment, K ³⁰ is H, and each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ^M is H, and R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, R ^3d is ClHbR ⁹ , and each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ^3d is CH2R ⁹ , and R ⁵ and R ⁶ , taken together, are

10 =CR ^c R ^d . In another aspect of this embodiment, R ^3d is alkenyl or substituted alkenyl, and each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ^M is alkenyl or substituted alkenyl, and R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, R ^3d is alkynyl or substituted alkynyl, and each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ^M is alkynyl or

15 substituted alkynyl, and R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, X ⁵ is CH and each X ⁶ and X ⁷ is N. In another aspect of this embodiment, X ⁵ is CH, each X ⁶ and X ⁷ is N, and R ⁷⁰ is -C(O)NR ^7b R ^7c . In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R" is not H. In another aspect of this

20 embodiment, each W ¹ and W ² is independently Y ² -R* wherein each Y ² is — O- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -N(R)- and R ^x is not H.

^• In another preferred embodiment of Formula IV, B ³ is

_X 6 _^ R ^{70 /Ύ 1ST}

wherein X ⁵ is CH, each X ⁶ and X ⁷ is N, R ^a is H, each R ¹⁰ is H and

90

5 L ¹ is O. In another aspect of this embodiment, R ^3d is H, and each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ³⁰ is H, and R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, R ^3d is CH2R ⁹ , and each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ^3d is CH2R ⁹ , and R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, R ³⁰ is alkenyl or

10 substituted alkenyl, and each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ^3d is alkenyl or substituted alkenyl, and R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, R ^3d is alkynyl or substituted alkynyl, and each R ^s and R ⁶ is H. In another aspect of this embodiment, R ^M is alkynyl or substituted alkynyl, and R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In

15 another aspect of this embodiment, R ⁷⁰ is -C(O)NR ⁷¹⁵ R ⁷ =.

In another preferred embodiment of Formula IV, B ³ is ,R ^{70 r r}

/ ^ wherein R ⁷⁰ is -C(O)NH ₂ , X ⁵ is CH, each X ⁶ and X ⁷ is N, R ^a is H, each R ¹⁰ is H and L ¹ is O. In another aspect of this embodiment, R ^3d is H, and each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ^M is H, and R ⁵ and

20 R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, R ^3d is CH2R ⁹ , and each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ³⁰ is CH2R ⁹ , and R ⁵ and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, R ^3d is alkenyl or substituted alkenyl, and each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ³⁰ is alkenyl or substituted alkenyl, and R ⁵

25 and R ⁶ , taken together, are =CR ^c R ^d . In another aspect of this embodiment, R ^3d is alkynyl or substituted alkynyl, and each R ⁵ and R ⁶ is H. In another aspect of this embodiment, R ^3d is alkynyl or substituted alkynyl, and R ⁵ and R ⁶ , taken together,

91

are =CR ^c R ^d . In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is independently -O- or -N(R)- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R ^x wherein each Y ² is -O- and R ^x is not H. In another aspect of this embodiment, each W ¹ and W ² is independently Y ² -R* wherein each Y ² is

10 independently -N(R)- and R ^x is not H.

The compounds of the Formulas I, II, III and IV bear a phosphonate group,

w ^; IL i which may be a prodrug moiety wherein W and W ² are each independently a group of the formula:

R ^x / λ Y ¹ _ _Y 2 p.

15 M2 wherein: each Y ² is independently a bond, O, CR ₂ , NR, ⁺ N(O)(R), N(OR), ⁺ N(O)(OR), N-NR ₂ , S, S-S, S(O), or S(O) ₂ ;

M2 is 0, 1 or 2;

20 each R ^y is independently H ₇ F, Cl, Br ₇ 1, OH, R, -Q=Y ¹ JR, -Q=Y ¹ JOR, - Q=Y^N(R) ₂ , -N(R) ₂ , - ⁺ N(R) ₃ , -SR, -S(O)R, -S(O) ₂ R, -S(O)(OR), -S(O) ₂ (OR), - OQ=YOR, -OC(=Y ¹ )OR, -OQ=Y ¹ XN(R) ₂ ), -SQ=Y ¹ JR, -SC(=Y ¹ )OR, -SQ=Y ¹ XN(R) ₂ ), -N(R)Q=Y ¹ JR, -N(R)Q=Y ¹ PR ₇ or -N(R)Q=Y ¹ JN(R) ₂ , amino (-NH ₂ ), ammonium (-NHb ⁺ ), alkylamino, dialkylamino, trialkylammonium, Ci-Cs alkyl, Ci-Cs

25 alkylhalide, carboxylate, sulfate, sulfamate, sulfonate, Ci-Cs alkylsulfonate, Ci-Cs

92

alkylainino, Ci-Cs alkylhydroxyl, Ci-Cs alkylthioϊ, alkylsulfone (-SO2R), sulfonamide (-SOzNRa), alkylsulfoxide (-SOR), ester (-C(O)OR), arrrido (-C(=O)NR2), nitrile (-CN), azido (-N ₃ ), nitro (-NO ₂ ), Ci-Cs alkoxy (-OR), Ci-Cs alkyl, Ci-Cs substituted alkyl, Cz-Cs alkenyl, Cz-Cs substituted alkenyl, C2-C8 alkynyl, C2-C8 substituted alkynyl, a protecting group or W ³ ; or when taken

10 together, two R ^y on the same carbon atom form a carbocyclic ring of 3 to 7 carbon atoms; each R ^x is independently R ^y , a protecting group, or the formula:

Y

M1a M12c M1c M1d wherein:

15 MIa, MIc, and MId are independently 0 or 1;

M12c is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12; or when taken together, two R ^x are optionally substituted C2-C4 alkylene thereby forming a phosphorous-containing heterocycle; each R is H, halogen, Ci-Cs alkyl, Ci-Cs substituted alkyl, Ci-Ce alkenyl,

20 C2-C8 substituted alkenyl, C2-C8 alkynyl, Gz-Cs substituted alkynyl, C6-C20 aryl, C6-C20 substituted aryl, C2-C20 heterocycle, C2—C20 substituted heterocycle, or a protecting group;

W ³ is W ⁴ or W ⁵ ; W ⁴ is R, -C(Y ¹ )R ^y , -C(Y ¹ JW ⁵ , -SO ₂ R ^y , or -SO ₂ W ⁵ ; and W ⁵ is a carbocycle or a heterocycle wherein W ⁵ is independently substituted with 0 to 3

25 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 comprising a mono- or bicyclic carbocycle or heterocycle. W ⁵ may have 3 to 10

93

5 ring atoms, e.g., 3 to 7 ring atoms. The W ⁵ rings are saturated when containing 3 ring atoms, saturated or mono-unsaturated when containing 4 ring atoms, saturated, or mono- or di-unsaturated when containing 5 ring atoms, and saturated, mono- or di~unsaturated, or aromatic when containing 6 ring atoms. A W ⁵ heterocycle may be a monocycle having 3 to 7 ring members (2 to 6

10 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

15 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 as 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 hetero atoms selected from N and S) arranged as a bicyclo [5,6] or [6,6] system. The W ⁵ heterocycle may be bonded to Y ² through a carbon, nitrogen, sulfur or other

20 atom by a stable covalent bond.

W ⁵ heterocycles include for example, pyridyl, dihydropyridyl 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:

25

94

N ' ¹ S

H

*— N H

^{¥ <} -N ^■ . ^ύ S-"* .and

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

PH

.Cl

OH Cl

N

NH ₂

10

NH NH I— N NH

\

I— { \ I— N SH _i→ ( \o ₂

Examples of substituted phenyl carbocycles include:

95

Embodiments of ^w of Formula I, II, III or IV compounds include substructures such as:

O

.ιλ ,R ^X

X υ ^2'

10 wherein Y ² is -O- or -N(R)-.

O

Another embodiment of ^" ^w of Formula I, II, m or IV includes the substructures:

JUY. ^S RV

-W ⁵

15 wherein Y ² is O, N(R) or S.

96

diment of ¹ Xf .α L ² J^ Another embo - - ^w ₂ of Formula I, II, III or IV compounds include the substructures:

wherein W ⁵ is a carbocycle such as phenyl or substituted phenyl. Such a substructure includes:

-LJ v° ^py

X L ^{2^} V OR

10 wherein Y ² is O or N(R) and the phenyl carbocycle is substituted with 0 to 3 R ^y groups.

An embodiment of ^ of Formula I, II, III or IV includes

15 phenyl phosphonamidate amino acids, e.g. alanate esters and phenyl phosphonate-lactate esters:

\ O CH ₃

OR V and

97 ^λ OR

5 The chiral carbon of the amino acid and lactate moieties may be either the

R or S configuration or the racemic mixture.

,.W ¹

Another embodiment of V ^W2 of Formula I, Formula II, Formula III, or Formula IV is

10 wherein Y ¹ is O or S and each Y ² is -O- or -N(R)-. In a preferred embodiment, W ¹ and W ² are independently nitrogen-linked naturally occurring amino acids or their enantiomers. In another preferred embodiment, W ¹ and W ² are independently naturally occurring 2-hydroxy carboxylic acids or their

15 enantiomers that are linked through the 2-hydroxy group.

98

^W ¹

Another embodiment of ¹ XT ^L- ^{^} W ² of Formula I, Formula II, Formula III, or Formula IV is

O

Il

*lA ,R

X -L ² '

^* R

In one preferred embodiment each R is independently Ci-Ce alkyl. In another preferred embodiment each R is independently C6-C20 aryl or C6-C20 substituted

10 aryl.

Embodiments of R* include esters, carbamates, carbonates, thioesters, amides, thioamides, and urea groups:

,R ^y

Cellular Accumulation

15 One aspect of the invention is HCV polymerase inhibitor compounds capable of accumulating in human PBMC (peripheral blood monocyte cells).

Optionally, the compounds of the invention demonstrate improved • intracellular half-life of the compounds or intracellular metabolites of the compounds in human PBMC when compared to analogs of the compounds not

20 having the phosphonate or phosphonate prodrug. Typically, the half-life is improved by at least about 50%, more typically at least in the range 50-100%, still more typically at least about 100%, more typically yet greater than about 100%.

99

5 In one embodiment, the intracellular half-life of a metabolite of the compound in human PBMC is improved when compared to an analog of the compound not having the phosphonate or phosphonate prodrug. In such embodiments, the metabolite is typically generated intracellularly, more typically, it is generated within Human PBMC. Still more typically, the

10 metabolite is a product of the cleavage of a phosphonate prodrug within human PBMCs. More typically yet, the phosphonate prodrug is cleaved to form a metabolite having at least one negative charge at physiological pH. Most typically, the phosphonate prodrug is enzymatically cleaved within human PBMC to form a phosphonate having at least one active hydrogen atom of the

15 form P-OH.

Recursive Substituents

Selected substituents within the compounds of the invention are present to a recursive degree. In this context, "recursive substituent" means that a

20 substituent may recite another instance of itself. Because of the recursive nature of such substituents, theoretically, a large number of compounds may be present in any given embodiment. For example, R* contains a R ^y substituent. R ^y can be R. One of ordinary skill in the art of medicinal chemistry understands that the total number of such substituents is reasonably limited by the desired properties

25 of the compound intended. Such properties include, by of example and not limitation, physical properties such as molecular weight, solubility or log P, application properties such as activity against the intended target, and practical properties such as ease of synthesis.

By way of example and not limitation, W ³ and R^ are recursive

30 substituents in certain embodiments. Typically, each of these may independently occur 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1,

100

5 or 0, times in a given embodiment. More typically, each of these may independently occur 12 or fewer times in a given embodiment. More typically yet, W ³ - will occur 0 to 8 times, R ^y will occur 0 to 6 times in a given embodiment. Even more typically, W ³ will occur 0 to 6 times, R>" will occur 0 to 4 times and R ³ will occur 0 to 8 times in a given embodiment.

10 Recursive substituents are an intended aspect of the invention. One of ordinary skill in the art of medicinal chemistry understands the versatility of such substituents. To the degree that recursive substituents are present in an embodiment of the invention, the total number will be determined as set forth above.

15 Protecting Groups

In the context of the present invention, embodiments of protecting groups include prodrug moieties and chemical protecting groups.

Protecting groups are available, commonly known and used, and are optionally used to prevent side reactions with the protected group during

20 synthetic procedures, i.e. routes or methods to prepare the compounds of the invention. For the most part the decision as to which groups to protect, when to do so, and the nature of the chemical protecting group "PRT" will be dependent upon the chemistry of the reaction to be protected against (e.g., acidic, basic, oxidative, reductive or other conditions) and the intended direction of the

25 synthesis. The PRT groups do not need to be, and generally are not, the same if the compound is substituted with multiple PRT. In general, PRT will be used to protect functional groups such as carboxyl, hydroxyl or amino groups and to thus prevent side reactions or to otherwise facilitate the synthetic efficiency. The order of deprotection to yield free, deprotected groups is dependent upon the

30 intended direction of the synthesis and the reaction conditions to be encountered, and may occur in any order as determined by the artisan.

101

5 Various functional groups of the compounds of the invention may be protection. For example, protecting groups for -OH groups (whether hydroxyl, carboxylic acid, phosphonic acid, or other functions) are embodiments of "ether- or ester-forming groups". Ether- or ester-forming groups are capable of functioning as chemical protecting groups in the synthetic schemes set forth

10 herein. However, some hydroxyl and thio protecting groups are neither ether- nor ester-forming groups, as will be understood by those skilled in the art, and are included with amides, discussed below.

A very large number of hydroxyl protecting groups and amide-forming groups and corresponding chemical cleavage reactions are described in

15 "Protective Groups in Organic Chemistry", Theodora W. Greene (John Wiley & Sons, Inc., New York, 1991, ISBN 0-471-62301-6) ("Greene"). See also Kocienski, Philip J.; "Protecting Groups" (Georg Thieme Verlag Stuttgart, New York, 1994), which is incorporated by reference in its entirety herein. In particular Chapter 1, Protecting Groups: An Overview, pages 1-20, Chapter 2, Hydroxyl Protecting

20 Groups, pages 21-94, Chapter 3, Diol Protecting Groups, pages 95-117, Chapter 4, Carboxyl Protecting Groups, pages 118-154, Chapter 5, Carbonyl Protecting Groups, pages 155-184. For protecting groups for carboxylic acid, phosphonic acid, phosphonate, sulfonic acid and other protecting groups for acids see Greene as set forth below. Such groups include by way of example and not

25 limitation, esters, amides, hydrazides, and the like.

Ether- and Ester-forming protecting groups

Ester-forming groups include: (1) phosphonate ester-forming groups, such as phosphonamidate esters, phosphorothioate esters, phosphonate esters, and phosphon-bis-amidates; (2) carboxyl ester-forming groups, and (3) sulphur 30 ester-forming groups, such as sulphonate, sulfate, and sulfinate.

The phosphonate moieties of the compounds of the invention may or may

102

5 not be prodrug moieties, i.e. they may or may not be susceptible to hydrolytic or enzymatic cleavage or modification. Certain phosphonate moieties are stable under most or nearly all metabolic conditions. For example, a dialkylphosphonate, where the alkyl groups are two or more carbons, may have appreciable stability in vivo due to a slow rate of hydrolysis.

10 Within the context of phosphonate prodrug moieties, a large number of structurally-diverse prodrugs have been described for phosphonic acids (Freeman and Ross in Progress in Medicinal Chemistry 34: 112-147 (1997) and are included within the scope of the present invention. An exemplary embodiment of a phosphonate ester-forming group is the phenyl carbocycle in a

15 substructure having the formula:

wherein the phenyl carbocycle is substituted with 0 to 3 R groups. Also, in this embodiment, where Y ² is O, a lactate ester is formed. Alternatively, where Y ² is NR, N-OR or N-N(R)2, then phosphonamidate esters result. R 20 susbstituents include H and C1-C12 alkyl.

In its ester-forming role, a protecting group typically is bound to any acidic group such as, by way of example and not limitation, a -CO2H or -C(S)OH group, thereby resulting in -CO2R* where R* is defined herein. Also, R ^x for example includes the enumerated ester groups of WO 95/07920. 25 Examples of protecting groups include (a)-(j):

(a) C3— C12 heterocycle (described above) or aryl. These aromatic groups optionally are poly cyclic or monocyclic. Examples include phenyl, spiryl, 2- and

103

5 3-pyrrolyl, 2- and 3-thienyl, 2- and 4-imidazolyi, H-, 4- and 5-oxazolyl, 3- and 4- isoxazolyl, 2-, 4- and 5-thiazolyl, 3-, 4- and 5-isothiazolyl, 3- and 4-pyrazolyl, 1-, 2-, 3- and 4-pyridinyl, and 1-, 2-, Ar and 5-pyrimidinyl.

(b) C3-C12 heterocycle or aryl substituted with halo, R ¹ , W-O-Q.\XL\2. alkylene, C1-C12 alkoxy, CN, NO2, OH, carboxy, carboxyester, thiol, thioester,

10 C1-C12 haloalkyl (1-6 halogen atoms), C2-C12 alkenyl or C2-C12 alkynyl. Such groups include, but are not limited to, 2-, 3- and 4-alkoxyphenyl (C1-C12 alkyl); 2-, 3- and 4-methoxyphenyl; 2-, 3- and 4-ethoxyphenyl; 2,3-, 2,4-, 2,5-, 2,6-, 3,4- and 3,5-diethoxyphenyl; 2- and 3-carboethoxy-4-hydroxyphenyl; 2- and 3- ethoxy-4-hydroxyphenyl; 2- and 3~ethoxy-5-hydroxyphenyl; 2- and 3-ethoxy-6-

15 hydroxyphenyl; 2-, 3- and 4-O-acetylphenyl; 2-, 3- and 4-dimethylaminophenyl; 2-, 3- and 4-methylmercaptophenyl; 2-, 3- and 4-halophenyl (including 2-, 3- and 4-fluorophenyl and 2-, 3- and 4-chlorophenyl); 2,3-, 2,4-, 2,5-, 2,6-, 3,4- and 3,5- dimethylphenyl; 2,3-, 2,4-, 2,5-, 2,6-, 3,4- and 3,5-biscarboxyethylphenyl; 2,3-, 2,4-, 2,5-, 2,6-, 3,4- and 3,5-dimethoxyphenyl; 2,3-, 2,4-, 2,5-, 2,6-, 3,4- and 3,5-

20 dihalophenyl (including 2,4-difluorophenyl and 3,5-difluorophenyl); 2-, 3- and 4- haloalkylphenyl (1 to 5 halogen atoms, C1-C12 alkyl including 4- trifluoromethylphenyl); 2-, 3- and 4-cyanophenyl; 2-, 3- and 4-nitrophenyl; 2-, 3- and 4-haloalkylbenzyl (1 to 5 halogen atoms, C1-C12 alkyl including 4- trifluoromethylbenzyl and 2-, 3- and 4-trichloromethylphenyl and 2-, 3- and 4-

25 trichloromethylphenyl); 4-N-methylpiperidinyl; 3-N-methylpiperidinyl; 1- ethylpiperazinyl; benzyl; alkylsalicylphenyl (C1-C4 alkyl, including 2-, 3- and 4- ethylsalicylphenyl); 2-,3- and 4-acetylphenyl; 1,8-dihydroxynaphthyl (-C10H6- OH) and aryloxy ethyl [Cό-Cg aryl (including phenoxy ethyl)], 2,2'- dihydroxybiphenyl; 2-, 3- and 4-N,N-dialkylaminophenol; -C6H4CH2-N(CH3)2;

30 trimethoxybenzyl; triethoxybenzyl; and 2-alkyl pyridinyl (Ci -4 alkyl).

104

-N -CH ₂ -O-C(O)- " ( _C ) O H . \=/ . ^ \==/ _{; C4} c ₈

esters of 2-carboxyphenyl; and C1-C4 alkylene-C3~C6 aryl (including benzyl, - CH2-pyrrolyl, -CH ₂ -thienyl, -CH2-imidazolyl _/ -CH2-oxazolyl, -CH2-isoxazolyl, - CH2-thiazolyl, -CH2-isothiazolyl, -CH2-pyrazolyl, ~CH ₂ -pyridinyl and -CH2- pyrimidinyl) substituted in the aryl moiety by 3 to 5 halogen atoms or 1 to 2 10 atoms or groups selected from halogen, C1-C12 alkoxy (including methoxy and ethoxy), cyano, nitro, OH, C1-C12 haloalkyl (1 to 6 halogen atoms; including - CH2CCI3), C1-C12 alkyl (including methyl and ethyl), C2-C12 alkenyl or C2-C12 alkynyl.

(d) Alkoxy ethyl [C1-C6 alkyl including -CH2-CH2-O-CH3 (methoxy 15 ethyl)], alkyl substituted by any of the groups set forth above for aryl, in particular OH or by 1 to 3 halo atoms (including -CH3, -CH(CH ₃ )2, -C(CH ₃ )3, - CH ₂ CH ₃ , -(CH ₂ )2CH ₃ , -(CH ₂ )3CH ₃ , -(CH ₂ UCH ₃ , -(CH ₂ )SCH^CH ₂ CH ₂ F, -

CH ₂ CH ₂ Cl, -CH ₂ CF ₃ , and -CH ₂ CCl ₃ ), ^N ' , -N-2- propylmorpholino, 2,3-dihydro-6-hydroxyindene, sesamol, catechol monoester, - 20 CH ₂ -C(O)-N(R ¹ )2, -CH ₂ -S(O)(R ¹ ), -CH2-S(O)2(R ¹ ), -CH ₂ -CH(OC(O)CH ₂ R ¹ )- CH ₂ (OC(O)CH ₂ R ¹ ), cholesteryl, enolpyruvate (HOOC-C(=CH ₂ )-) or glycerol.

(e) A 5 or 6 carbon monosaccharide, disaccharide or oligosaccharide (3 to 9 monosaccharide residues).

(f) Triglycerides such as α-D-(3-diglycerides (wherein the fatty acids 25 composing glyceride lipids generally are naturally-occurring saturated or unsaturated Cβ-26/ C6-18 or Cό-10 fatty acids such as linoleic, lauric, myristic, palmitic, stearic, oleic, palmitoleic, linolenic and the like fatty acids) linked to

105

5 acyl of the parental compounds herein through a glyceryl oxygen of the triglyceride;

(g) Phospholipids linked to the carboxyl group through the phosphate of the phospholipid.

(h) Phthalidyl (shown in Fig. 1 of Clayton et al., Antimicrob. Agents Chemo. 10 (1974) 5(6):670-671.

(i) Cyclic carbonates such as (5-Rd-2-oxo~l,3-dioxolen-4-yl) methyl esters (Sakamoto et al _v Client. Pharm. Bull. (1984) 32(6)2241-2248) where Rd is Ri, R4 or aryl.

-CH ₂ C(O)N p

15

The hydroxy! groups of the compounds of this invention optionally are substituted with one of groups III, IV or V disclosed in WO 94/21604, or with isopropyl.

As further embodiments, Table A lists examples of protecting group ester

20 moieties that for example can be bonded via oxygen to -C(O)O- and -P(O)(O-)2 groups. Several amidates also are shown, which are bound directly to -C(O)- or - P(O)2- Esters of structures 1-5, 8-10 and 16, 17, 19-22 are synthesized by reacting the compound herein having a free hydroxyl with the corresponding halide (chloride or acyl chloride and the like) and N ,N-dicyclohexyl-N-morpholine

25 carboxamidine (or another base such as DBU, triethylamine, CsCO3, N,N- dimethylaniline and the like) in DMF (or other solvent such as acetonitrile or N- methylpyrrolidone). When the compound to be protected is a phosphonate, the esters of structures 5-7, 11, 12, 21, and 23-26 are synthesized by reaction of the alcohol or alkoxide salt (or the corresponding amines in the case of compounds

30 such as 13, 14 and 15) with the monochlorophosphonate or dichlorophosphonate

106

5 (or another activated phosphonate).

Table A

1. -CH2-C(O)-N(Ri)2 * 10. -CH ₂ -O-C(O)-C(CH3)3 10 2. -CH2-S(O)(Ri) 11. -CH2-CCI3

3. -CH ₂ -S(O) ₂ (Rl) 12. -C6H5

4. -CH ₂ -O-C(O)-CH ₂ -CeHs 13. -NH-CH2-C(O)O-CH ₂ CH3

5. 3-cholesteryl 14. -N(CH3)-CH ₂ -C(O)O-CH ₂ CH3

6. 3-pyridyl 15. -NHRi

15 7. N-ethylmorpholino 16. -CH ₂ -O-C(O)-CiOHi ₅

8. -CH ₂ -O-C(O)-C6H5 17. -CH ₂ -O-C(O)-CH(CH3) ₂

9. -CH ₂ -O-C(O)-CH ₂ CH3 18. -CH ₂ -CfH(OC(O)CH ₂ Ri)-CH ₂ - (OC(O)CH ₂ Ri)*

HO

-CH ₂ C(O)N^JD

20 19. 20. O H 21. HO

CH ₃ O(O)C

N

-CH ₂ -O-C(O) -\3 -CH ₂ CH ₂ -^ ^ / λ

22. 23. 24.

OCH,

CH ₃ CH ₂ O(O)C -CH ₂ OCH,

25. 26. OCHo

25 # - chiral center is (R), (S) or racemate.

107

Other esters that are suitable for use herein are described in EP 632048. Protecting groups also includes "double ester" forming profunctionalities. such as

-CH2θC(O)OCH3, o -CH2SCOCH3, -CH2θCON(CH3)2, or alkyl- or aryl-acyloxyalkyl groups of the structure -CH(R ¹ or W ⁵ )O((CO)R ³⁷ ) or

10 -CH(R ¹ or W ⁵ X(CO)OR ³⁸ ) (linked to oxygen of the acidic group) wherein R ³⁷ and R ³⁸ are alkyl, aryl, or alkylaryl groups (see U.S. Patent No.4,968,788). Frequently R ³⁷ and R ³⁸ are bulky groups such as branched alkyl, ortho-substituted aryl, meta-substituted aryl, or combinations thereof, including normal, secondary, iso- and tertiary alkyls of 1-6 carbon atoms. An example is the

15 pivaloyloxymethyl group. These are of particular use with prodrugs for oral administration. Examples of such useful protecting groups are alkylacyloxymethyl esters and their derivatives, including -

v. ,CX

CH(CH2CH2θCH3)OC(O)C(CH3)3, °

-CH ₂ OC(O)CiOHi ₅ , -CH ₂ OC(O)C(CH ₃ )3, -CH(CH ₂ OCH ₃ )OC(O)C(CH3)3, 20 -CH(CH(CH3)2)OC(O)C(CH3)3, -CH ₂ OC(O) CH ₂ CH(CH3>2,

-CH ₂ OC(O)COHH, -CH ₂ OC(O)CoHs -CH ₂ OC(O)CI ₀ HI ₅ ,

-CH ₂ OC(O)CH ₂ CH3, ~CH ₂ OC(O)CH(CH ₃ )2,

-CH ₂ OC(O)C(CH3)3 and -CH ₂ OC(O)CH ₂ CoH ₅ .

For prodrug purposes, the ester typically chosen is one heretofore used 25 ' for antibiotic drugs, in particular the cyclic carbonates, double esters, or the phthalidyl, aryl or alkyl esters.

In some embodiments the protected acidic group is an ester of the acidic group and is the residue of a hydroxyl-containing functionality. In other

108

5 " embodiments, an amino compound is used to protect the acid functionality. The residues of suitable hydroxyl or amino-containing functionalities are set forth above or are found in WO 95/07920. Of particular interest are the residues of amino acids, amino acid esters, polypeptides, or aryl alcohols. Typical amino acid, polypeptide and carboxyl-esterified amino acid residues are described on

10 pages 11-18 and related text of WO 95/07920 as groups Ll or L2. WO 95/07920 expressly teaches the amidates of phosphonic acids, but it will be understood that such amidates are formed with any of the acid groups set forth herein and the amino acid residues set forth in WO 95/07920.

Typical esters for protecting acidic functionalities are also described in

15 WO 95/07920, again understanding that the same esters can be formed with the acidic groups herein as with the phosphonate of the '920 publication. Typical ester groups are defined at least on WO 95/07920 pages 89-93 (under R ³¹ or R ³⁵ ), the table on page 105, and pages 21-23 (as R). Of particular interest are esters of unsubstituted aryl such as phenyl or arylalkyl such benzyl, or hydroxy-, halo-, 20 alkoxy-, carboxy- and/or alkylestercarboxy-substituted aryl or alkylaryl, especially phenyl, ortho-ethoxyphenyl, or C1-C4 allcylestercarboxyphenyl

(salicylate C1-C12 alkylesters).

The protected acidic groups, particularly when using the esters or amides of WO 95/07920, are useful as prodrugs for oral administration. However, it is

25 not essential that the acidic group be protected in order for the compounds of this invention to be effectively administered by the oral route. When the compounds of the invention having protected groups, in particular amino acid amidates or substituted and unsubstituted aryl esters are administered systemically or orally they are capable of hydrolytic cleavage in vivo to yield the

30 free acid.

A plurality of the acidic hydroxyls may be protected. If more than one

109

5 acidic hydroxyl is protected then the same or a different protecting group is employed, e.g., the esters may be different or the same, or a mixed amidate and ester may be used.

Typical acid hydroxy protecting groups described in Greene (pages 14- 118) include substituted methyl and alkyl ethers, substituted benzyl ethers, silyl . 10 ethers, esters including sulfonic acid esters, and carbonates. For example:

• Ethers (methyl, f-butyl, allyl);

• Substituted Methyl Ethers (Methoxymethyl, Methylthiomethyl, t- Butylthiomethyl, (Phenyldimethylsilyl)methoxymethyl, Benzyloxymethyl, p- Methoxybenzyloxymethyl, (4-Methoxyphenoxy)methyl, Guaiacolmethyl, t-

15 Butoxymethyl, 4-Pentenyloxymethyl, Siloxymethyl, 2-Methoxyethoxymethyl,

2,2,2-Trichloroethoxymethyl, Bis(2-chloroethoxy)methyl, 2- (Trimethylsilyl)ethoxymethyl, Tetrahydropyranyl, 3-

Bromotetrahydropyranyl, Tetrahydropthiopyranyl, 1-Methoxycyclohexyl, 4- Methoxytetrahydropyranyl, 4-Methoxytetrahydrothiopyranyl, 4-

20 Methoxy tetrahydropthiopyranyl S,S-Dioxido, l-[(2-Chloro-4-methyl)phenyl]-

4-methoxypiperidin-4-yl, l,4-Dioxan-2-yl, Tetrahydrofuranyl, Tetrahydrothiofuranyl, 2,3,3a _/ 4,5,6,7,7a-Octahydro-7 _/ 8,8-trimethyl-4,7- methanobenzofuran-2-yl));

• Substituted Ethyl Ethers (1-Ethoxy ethyl, l-(2-Chloroethoxy)ethyl, 1-Methyl- 25 1-methoxyethyl, 1-Methyl-l-benzyloxyethyl, l-Methyl-l-benzyloxy-2- fluoroethyl, 2,2,2-Trichloroethyl, 2-Trimethylsilylethyl, 2- (Phenylselenyl)ethyl,

• p-Chlorophenyl, p-Methoxyphenyl, 2,4-Dinitrophenyl, Benzyl);

• Substituted Benzyl Ethers (p-Methoxybenzyl, 3,4-Dimethoxybenzyl, o- 30 Nitrobenzyl, p-Nitrobenzyl, /7-Halobenzyl, 2,6-Dichlorobenzyl, p-

Cyanobenzyl, p-Phenylbenzyl, 2- and 4-Picolyl, 3-Methyl-2-picolyl N-Oxido, Diphenylmethyl, p,p'-Dinitrobenzhydryl, 5-Dibenzosuberyl, Triphenylirtethyl,

110

5 α-Naphthyldiphenylmethyl, p-methoxyphenyldiphenylmethyl, Oi(p- methoxyphenyl)phenylmethyl, Tri(p-methoxyphenyl)methyl, 4-(4'- Bromophenacyloxy)phenyldiphenylmethyl, 4,4',4"-Tris(4,5- dichlorophthalimidophenyl)methyl, 4,4',4"-Tris(levulinoyloxyphenyl)iiιethyl, 4,4^4"-Tris(benzoyloxyphenyl)methyl _/ 3-(IInidazol-l-ylmethyl)bis(4',4' ¹ - 10 dimethoxyphenyl)methyl, l,l-Bis(4-methoxyphenyl)-l'-pyrenyhnethyl, 9-

Anthryl, 9-(9-Phenyl)xanthenyl, 9-(9-Phenyl-10-oxo)anthryl, 1,3- Benzodithiolan-2-yl, Benzisothiazolyl S,S-Dioxido);

• Silyl Ethers (Trirrvethylsilyl, Triethylsilyl/ Triisopropylsilyl, Dimethylisopropylsilyl, Diethylisopropylsilyl, Dimethylthexylsilyl, t-

15 Butyldimethylsilyl, t-Butyldiphenylsilyl, Tribenzylsilyl, Tri-p-xylylsilyl,

Triphenylsilyl, Diphenylmethylsilyl, t-Butylmethoxyphenylsilyl);

• Esters (Formate, Benzoylformate, Acetate, Choroacetate, Dichloroacetate, Trichloroacetate, Trifluoroacetate, Methoxyacetate, Triphenylmethoxyacetate, Phenoxyacetate, p-Chlorophenoxyacetate, p-poly-Phenylacetate, 3-

20 Phenylpropionate, 4-Oxopentanoate (Levulinate), 4,4-

(Ethylenedithio)pentanoate, Pivaloate, Adamaritoate, Crotonate, 4- Methoxycrotonate, Benzoate, p-Phenylbenzoate, 2,4,6-Trimethylbenzoate (Mesitoate));

• Carbonates (Methyl, 9-Fluorenylmethyl, Ethyl, 2,2,2-Trichloroethyl, 2-

25 (Trimethylsilyl)ethyl, 2-(Phenylsulionyl)ethyl, 2-(Triphenylphosphonio)ethyl,

Isobutyl, Vinyl, Allyl, p-Nitrophenyl, Benzyl, p-Methoxybenzyl, 3,4- Dimethoxybenzyl, o-Nitrobenzyl, p-Nitrobenzyl, S-Benzyl Thiocarbonate, 4- Ethoxy-1-naphthyl, Methyl Dithiocarbonate);

.• Groups With Assisted Cleavage (2-Iodobenzoate, 4-Azidobutyrate, 4-Nitro-4- 30 methylpentanoate, o-(Dibromomethyl)benzoate, 2-Formylbenzenesulfonate,

2-(Methylthiomethoxy)ethyl Carbonate, 4-(Methylthiomethoxy)butyrate, 2- (Methylthiomethoxymethyl)benzoate); Miscellaneous Esters (2,6-Dichloro-4-

111

5 methylphenoxyacetate, 2,6-Dichloro-4-(l, 1,3,3 tetramethylbutyl)phenoxyacetate, 2,4-Bis(l _/ l-dimethylpropyl)phenoxyacetate, Chlorodiphenylacetate, Isobutyrate, Monosuccinate, (£)-2-Methyl-2- butenoate (Tigloate), o-(Methoxycarbonyl)benzoate, p-poly-Benzoate, α- Naphthoate, Nitrate, Alkyl N,N,N',N'-Tetramethylphosphorodiamidate, N- 10 Phenylcarbamate, Borate, Dimethylphosphinothioyl, 2,4-

Dinitrophenylsulfenate); and

• Sulfonates (Sulfate, Methanesulfonate (Mesylate), Benzylsulfonate, Tosylate). Typical 1,2-diol protecting groups (thus, generally where two OH groups are taken together with the protecting functionality) are described in Greene at 15 pages 118-142 and include Cyclic Acetals and Ketals (Methylene, Ethylidene, 1-t- Butylethylidene, 1-Phenylethylidene, (4-Methoxyphenyl)ethylidene, 2,2,2- Trichloroethylidene, Acetonide (Isopropylidene), Cyclopentylidene, Cyclohexylidene, Cycloheptylidene, Benzylidene, p-Methoxybenzylidene, 2,4- Dimethoxybenzylidene, 3,4-Dimethoxybenzylidene, 2-Nitrobenzylidene); Cyclic 20 Ortho Esters (Methoxymethylene, Ethoxymethylene, Dimethoxymethylene, 1- Methoxy ethylidene, 1-Ethoxyethylidine, 1,2-Dimethoxyethylidene, α- Methoxybenzylidene, 1-(N _/ N-Dimethylamino)ethylidene Derivative, α -(N ,N- Dimethylamino)benzylidene Derivative, 2-Oxacyclopentylidene); Silyl Derivatives (Di-t-butylsilylene Group, 1,3-(1,1,3,3-

25 Tetraisopropyldisiloxanylidene), and Tetra-t-butoxydisiloχane-l,3-diylidene), Cyclic Carbonates, Cyclic Boronates, Ethyl Boronate and Phenyl Boronate.

More typically, 1,2-diol protecting groups include those shown in Table B, still more typically, epoxides, acetonides, cyclic ketals and aryl acetals.

30

112

Table B

V V °γ° V° V° o o o o

\'° R'O-- ^N V ^{O R9} °-N, jo ^R9Q -^ P RO' ^{^} o I _O A _{O R9o} λ _o wherein R ⁹ in Table B is Cl -Co alkyl. Amino protecting groups

Another set of protecting groups include any of the typical amino 10 protecting groups described by Greene at pages 315-385. They include:

• Carbamates: (methyl and ethyl, 9-fluorenylmethyl, 9(2-sulfo)fluorenylmethyl _/ 9-(2 _/ 7-dibromo)fluorenylmethyl 2,7-di-t-butyl-[9-(10,10-dioxo-10 _/ 10 _/ 10 _/ 10- tetrahydrotWoxanthyl^methyl, 4-methoxyphenacyl);

• Substituted Ethyl: (2,2,2-trichoroethyl, 2-trimethylsilylethyl, 2-ρhenylethyl, 1- 15 (l-adamantylj-l-methylethyl l^-dimethyl^-haloethyl, l,l-dimethyl-2,2- dibromoethyl, lj-dimethyl^^^-trichloroethyl, 1 -methyl- 1 -(4- biphenylyl)ethyl, l-(3,5-di-f-butylphenyl)-l-methylethyl, 2-(2'- and 4'- pyridyl)ethyl, 2-(N _/ N-dicyclohexylcarboxamido)ethyl, i-butyl, 1-adamantyl, vinyl, allyl, 1-isopropylallyl, cinnamyl, 4-nitrocinnamyl, 8-quinolyl, N- 20 hydroxypiperidinyl, alkyldithio, benzyl, /?-methoxybenzyl, p-nitrobenzyl, γ- bromobenzyl, p-chlorobenzyl, 2,4-dichlorobenzyl, 4-methylsulfinylbenzyl, 9- anthrylmethyl, diphenylmethyl);

• Groups With Assisted Cleavage: (2-methylthioethyl, 2-methylsulfonylethyl, 2-(p-toluenesulfonyl)ethyl, [2-(l,3-dithianyl)]methyl, 4-methylthiophenyl, 2,4-

25 dimethylthiophenyl, 2-ρhosphonioethyl, 2-triρhenylphosphonioisopropyl,

113

5 l,l-dimethyI-2-cyanoethyl, m-choro-p-acyloxybenzyl, γ-

(dihydroxyboryl)benzyl, 5-benzisoxazolylmethyl, 2-(trifluoromethyl)-6- chromonylmethyl);

• Groups Capable of Photolytic Cleavage: (m-nitrophenyl, 3,5- dimethoxybenzyl, o-nitrobenzyl, 3,4-dimethoxy-6-nitrobenzyl, phenyl(o-

10 nitrophenyl)methyl); Urea-Type Derivatives (phenothiazinyl-(l O)-CaAoHyI,

N'-p-toluenesulfonylarrrinocarbonyl, N'-phenylaminothiocarbonyl);

• Miscellaneous Carbamates: (i-arnyl, S-benzyl thiocarbamate, p-cyanobenzyl, cyclobutyl, cyclohexyl, cyclopentyl, cyclopropylmethyl, p-decyloxybenzyl, diisopropylmethyl, 2,2-diinethoxycarbonylvinyl, o-(N,N-

15 dimethylcarboxamido)benzyl _/ l,l-dimethyl-3-(N,N- dimethylcarboxamidojpropyl, 1,1-dimethylpropynyl, di(2-pyridyl)methyl, 2- furanylmethyl, 2-Iodoethyl, Isobornyl, Isobutyl, Isonicotinyl, p-(p'- Methoxyphenylazo)benzyl, 1-methylcyclobutyl, 1-methylcyclohexyl, 1- methyl-1-cyclopropylmethyl, l-methyl-l-(3 _/ 5-dimethoxyphenyl)ethyl _/ 1-

20 methyl-l-(p-phenylazophenyl)ethyl, 1-methyl-l-phenylethyL l-methyl-l-(4- pyridyl)ethyl, phenyl, ^-(phenylazoybenzyl, 2,4,6-tri-f-butylphenyl, 4- (trirn.ethylarnrnoniurn)benzyl, 2,4,6-trimethylbenzyl);

• Amides: (N-formyl, N-aceryl, N-choroacetyl, N-trichoro acetyl, N- trifluoroacetyl, N-phenylacetyl, N-3-phenylpropionyl, N-picolinoyl, N-3-

25 pyridylcarboxamide, N-benzoylphenylalanyl/ N-benzoyl, N-p- phenylbenzoyl);

• Amides With Assisted Cleavage: (N-o-nitrophenylacetyl, N-o- nitrophenoxyacetyl, N-acetoacetyl, (N'-dithiobenzyloxycarbonylamino)acetyl, N-3-(p-hydroxyphenyl)propionyl, N-3-(o-nitrophenyl)propionyl, N-2-methyl-

30 2-(o-nitrophenoxy)propionyl _/ N-2-methyl-2-(o-phenylazophenoxy)propionyl,

N-4-chlorobutyryl, N-3-methyl-3-nitrobutyryl, N-o-nitrocinnamoyl, N- acetylmethionine, N-o-nitrσbenzoyl, N-o-(benzoyloxymethyl)benzoyl, 4,5-

114

5 diphenyl-3-oxazolin-2-one);

• Cyclic Imide Derivatives: (N-phthalimide, N~dithiasuccinoyl, N-2,3- diphenylmaleoyl, N-2,5-dimethylpyrrolyl, N-1,1,4,4- tetramethyldisilylazacyclopentane adduct, 5-substituted l _/ S-dimethyl-lAS- triazacyclohexan-2-one, 5-substituted l,3-dibenzyl-l,3-5-triazacycIohexan-2-

10 one, 1-substituted 3,5-dinitro-4-pyridonyl);

• N-Alkyl and N-Aryl Amines: (N-methyl, N-allyl, N-[2- (trimemylsilyl)ethoxy]methyl, N-3-acetoxypropyl, N-(l-isopropyl-4-nitro-2- oxo-3-pyrrolin-3-yl), Quaternary Ammonium Salts, N-benzyl, N-di(4- methoxyphenyl)methyl, N-5-dibenzosuberyl, N-triphenylmethyl, N-(4-

15 methoxyphenyl)diphenylmethyl, N-9-phenylfluorenyl, N-2,7-dichloro-9- fluorenylmethylene, N-ferrocenylmethyl, N-2-picolylamine N'-oxide);

• Imine Derivatives: (N-l,l-dimethylthiomethylene, N-benzylidene, N-p- methoxybenylidene, N-diphenylmethylene, N-[(2-pyridyl)mesityl]methylene, N^N^N'-dimethylaminomethylene, N,N'-isopropylidene, N-p-

20 nitrobenzylidene, N-salicylidene, N-5-chlorosalicylidene, N-(5-chloro-2- hydroxyphenyl)phenylmethylene, N-cyclohexylidene);

• Enamine Derivatives: (N-tS _/ δ-dimethyl-S-oxo-l-cyclohexenyl));

• N-Metal Derivatives (N-borane derivatives, N-diphenylborinic acid derivatives, N- [phenyl (pentacarbonylchromium- or -tungsten)]carbenyl, N-

25 copper or N-zinc chelate);

• N-N Derivatives: (N-nitro, N-nitroso, N-oxide);

• N-P Derivatives: (N-diphenylphosphinyl, N-dimethylthiophosphinyl, N- diphenylthiophosphinyl, N-dialkyl phosphoryl, N-dibenzyl phosphoryl, N- diphenyl phosphoryl);

30 • N-Si Derivatives, N-S Derivatives, and N-SuIf enyl Derivatives: (N- benzenesulfenyl, N-o-nitrobenzenesulfenyl, N-2,4-dinitrobenzenesulfenyl, N- pentachlorobenzenesulfenyl, N-2-nitro-4-methoxybenzenesulfenyl, N-

115

5 triphenylmethylsulfenyl, N-3-nitropyridinesulfenyl); and N-sulfonyl

Derivatives (N-p-toluenesulfonyl, N-benzenesulfonyl, N-2,3,6-trimethyl-4- methoxybenzenesulfonyl, N-2,4,6-trimethoxybenzenesulfonyl, N-2,6- dimethyl-4-methoxybenzenesulfonyl _/ N-pentamethylbenzenesulfonyl _/ N- 2,3,5,6,-tetramethyl-4~methoxybenzenesulfonyl, N-4-

10 methoxybenzenesulfonyl, N~2,4,6-trimethylbenzenesulfonyl, N-2,6- dimethoxy-4-methylbenzenesulfonyl, N^^^^^-pentamethylchroman-ό- sulfonyl, N-methanesulfonyl, N-β-trimethylsilyethanesulfonyl, N-9- anthracenesulfonyl, N-4-(4' _/ 8'-dimethoxynaphthylmethyl)benzenesulfonyl, N-benzylsulfonyl, N-trifluoromethylsulfonyl, N-phenacylsulfonyl).

15 Protected amino groups include carbamates, amides and amidines, e.g.

-NHC(O)OR ¹ , -NHC(O)R ¹ or -N=CR ¹ N(R ¹ )^ Another protecting group, also useful as a prodrug for amino or -NH(R ⁵ ), is:

O cAo Q

W ₆ ^N -— O

See for example Alexander, J. et al (1996) J. Med. Chem.39:480-486.

20 Amino acid and polypeptide protecting group and conjugates

An amino acid or polypeptide protecting group of a compound of the invention has the structure R ^15a NHCH(R ^16a )C(O)-, where R ^15a is H, an amino acid or polypeptide residue, or R ^15a , and R ^16a is defined below.

R ^16a is lower alkyl or lower alkyl (Cχ-C6) substituted with amino, carboxyl, 25 amide, carboxyl ester, hydroxyl, C6-C7 aryl, guanidinyl, imidazolyl, indolyl, sulfhydryl, sulfoxide, and/or alkylphosphate. R ^16a also is taken together with the amino acid α-N to form a proline residue (R ¹⁶ = -CH2)3-)- However, R ¹⁶ is generally the side group of a naturally-occurring amino acid such as H, -CH3, -

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5 CH(CH3)2, -CH2-CH(CH3)2, -CHCH3-CH2-CH3, -CH2-C6H5, -CH2CH2-S-CH3, -CH2OH, -CH(OH)-CH3, -CH2-SH, -CH2-C6H4OH, -CH2-CO-NH2, -CH2-CH2-CO-NH2, -CH2-COOH, -CH2-CH2-COOH, -(CH2)4~NH2 and - (CH2)3-NH-C(NH2)-NH2. R ^16a also includes l-guanidinoprop-3-yl, benzyl, 4- hydroxybenzyl, imidazol-4-yl, indol-3-yl, methoxyphenyl and ethoxyphenyl.

10 Another set of protecting groups include the residue of an amino- containing compound, in particular an amino acid, a polypeptide, a protecting group, -NHSO2R, NHC(O)R, -N(R)2, NH2 or -NH(R)(H), whereby for example a carboxylic acid is reacted, i.e. coupled, with the amine to form an amide, as in C(O)NR-. A phosphonic acid may be reacted with the amine to form a

15 phosphonamidate, as in -P(O)(OR)(NRz).

Amino acids have the structure R ^17c C(O)CH(R ^16a )NH-, where R ^17c is -OH, -OR, an amino acid or a polypeptide residue. Amino acids are low molecular weight compounds, on the order of less than about 1000 MW and which contain at least one amino or imino group and at least one carboxyl group. Generally

20 the amino acids will be found in nature, i.e., can be detected in biological material such as bacteria or other microbes, plants, animals or man. Suitable amino acids typically are alpha amino acids, Le. compounds characterized by- one amino or imino nitrogen atom separated from the carbon atom of one carboxyl group by a single substituted or unsubstituted alpha carbon atom. Of

25 particular interest are hydrophobic residues such as mono-or di-alkyl or aryl amino acids, cycloalkylamino acids and the like. These residues contribute to cell permeability by increasing the partition coefficient of the parental drug. Typically, the residue does not contain a sulfhydryl or guanidino substituent. Naturally-occurring amino acid residues are those residues found

30 naturally in plants, animals or microbes, especially proteins thereof.

Polypeptides most typically will be substantially composed of such naturally-

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5 occurring amino acid residues. These amino acids are glycine, alanine, valine, leucine, isoleucine, serine, threonine, cysteine, methionine, glutamic acid, aspartic acid, lysine, hydroxylysine, arginine, histidine, phenylalanine, tyrosine, tryptophan, proline, asparagine, glutamine and hydroxyproline. Additionally, unnatural amino acids, for example, valanine, phenylglycine and homoarginine

10 are also included. Commonly encountered amino acids that are not gene- encoded may also be used in the present invention. AU of the amino acids used in the present invention may be either the D- or L- optical isomer. In addition, other peptidomimetics are also useful in the present invention. For a general review, see Spatola, A. F., in Chemistry and Biochemistry of Amino Acids, Peptides

15 and Proteins, B. Weinstein, eds., Marcel Dekker, New York, p. 267 (1983).

When protecting groups are single amino acid residues or polypeptides they optionally are substituted with substituents. These conjugates are generally produced by forming an amide bond between a carboxyl group of the amino acid (or C-terminal amino acid of a polypeptide for example). Generally, only

20 one of any site in the scaffold drug-like compound is amidated with an amino acid as described herein, although it is within the scope of this invention to introduce amino acids at more than one permitted site. Usually, a carboxyl group of R ³ is amidated with an amino acid. In general, the ct-amino or α- carboxyl group of the amino acid or the terminal amino or carboxyl group of a

25 polypeptide are bonded to the scaffold, parental functionalities. Carboxyl or amino groups in the amino acid side chains generally may be used to form the amide bonds with the parental compound or these groups may need to be protected during synthesis of the conjugates as described further below.

With respect to the carboxyl-containing side chains of amino acids or

30 polypeptides it will be understood that the carboxyl group optionally will be blocked, e.g. esterified or amidated with R.

Such ester or amide bonds with side chain amino or carboxyl groups, like

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5 the esters or amides with the parental molecule, optionally are hydrolyzable in vivo or in vitro tinder acidic (pH <3) or basic (pH >10) conditions. Alternatively, they are substantially stable in the gastrointestinal tract of humans but are hydrolyzed enzymatically in blood or in intracellular environments. The esters or amino acid or polypeptide amidates also are useful as intermediates for the 10 preparation of the parental molecule containing free amino or carboxyl groups. The free acid or base of the parental compound, for example, is readily formed from the esters or amino acid or polypeptide conjugates of this invention by conventional hydrolysis procedures.

When an amino acid residue contains one or more chiral centers, any of 15 the D, L, meso, threo or erythro (as appropriate) racemates, scalemates or mixtures thereof may be used. In general, if the intermediates are to be hydrolyzed non-enzymatically (as would be the case where the amides are used as chemical intermediates for the free acids or free amines), D isomers are useful. On the other hand, L isomers are more versatile since they can be susceptible to 20 both non-enzymatic and enzymatic hydrolysis, and are more efficiently transported by amino acid or dipeptidyl transport systems in the gastrointestinal tract.

Examples of suitable amino acids whose residues are represented by R ^x or R ^y include the following: 25 Glycine;

Aminopolycarboxylic acids, e.g., aspartic acid, β-hydroxyaspartic acid, glutamic acid, β -hydroxyglutamic acid, β-methylaspartic acid, β- methylglutamic acid, β, β-dimethylaspartic acid, γ-hydroxyglutamic acid, β, γ- dihydroxyglutamic acid, β -phenylglutamic acid, γ-methyleneglutamic acid, 3- 30 aminoadipic acid, 2-aminopimelic acid, 2-aminosuberic acid and 2-aminosebacic acid;

Amino acid amides such as glutamine and asparagine;

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5 Polyamino- or polybasic-monocarboxylic acids such as arginine, lysine, β

-ammoalanine, γ -aminobutyrine, ornithine, citruline, homoarginine, homocitrulline, hydroxylysine, allohydroxylsine and diaminobutyric acid; Other basic amino acid residues such as histidine; Diaminodicarboxylic acids such as α, α'-diaminosuccinic acid, α, α'- 10 diaminoglutaric acid, α, α'-diaminoadipic acid, α, α'-diaminopimelic acid, α, α'- diamino- β-hydroxypimelic acid, a, α'-diaminosuberic acid, a, α'-diaminoazelaic acid, and α, α'-diaminosebacic acid;

Imino acids such as proline, hydroxyproline, allohydroxyproline, γ- methylproline, pipecolic acid, 5-hydroxypipecolic acid, and azetidine-2- 15 carboxylic acid;

A mono- or di-alkyl (typically Cχ-C8 branched or normal) amino acid such as alanine, valine, leucine, allylglycine, butyrine, norvaline, norleucine, heptyline, α-methylserine, α-amino-α-methyl-γ-hydroxyvaleric acid, α-amino- α-methyl-δ-hydroxyvaleric acid, α-amino- α-methyl-ε-hydroxycaproic acid, 20 isovaline, α-methylglutamic acid, α-aminoisobutyric acid, α-aminodiethylacetic acid, α-aminodiisopropylacetic acid, α-aminodi-n-propylacetic add, α- aminodiisobutylacetic acid, α-aminodi-n-butylacetic acid, α- aminoethylisopropylacetic acid, α-amino-n-propylacetic acid, a- aminodiisoamyacetic acid, α-methylaspartic acid, α-methylglutamic acid, 1- 25 aminocyclopropane-1-carboxylic acid, isoleucine, alloisoleucine, teri-leucine, β- methyltryptophan and α-amino- β-ethyl-β-phenylpropionic acid; β-phenylserinyl;

Aliphatic α-amino- β-hydroxy acids such as serine, β-hydroxyleucine, β- hydroxynorleucine, β -hydroxynorvaline, and α-amino-β-hydroxystearic acid; 30 α- Amino, a-, γ-, δ- or ε-hydroxy acids such as homoserine, δ - hydroxynorvaline, γ-hydroxynorvaline and ε-hydroxynorleucine residues; canavine and canaline; γ -hydroxyornithine;

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5 2-hexosaminic acids such as D-glucosaminic acid or D-galactosaminic acid; α-Amino-β-thiols such as penicillamine, β-thiolnorvaline or β- thiolbutyrine;

Other sulfur containing amino acid residues including cysteine; 10 homocystine, β-phenylmethionine, methionine, S-allyl-L-cysteine sulfoxide, 2- thiolhistidine, cystathionine, and thiol ethers of cysteine or homocysteine;

Phenylalanine, tryptophan and ring-substituted α-amino acids such as the phenyl- or cyclohexylamino acids α-aminophenylacetic acid, Oi- aminocyclohexylacetic acid and α-amino-β-cyclohexylpropionic acid; 15 phenylalanine analogues and derivatives comprising aryl, lower alkyl, hydroxy, guanidino, oxyalkylether, nitro, sulfur or halo-substituted phenyl (e.g., tyrosine, methyltyrosine and o-chloro-, p-chloro-, 3,4-dichloro, o- _r m- or p-methyl-, 2,4,6- trimethyl-, 2-ethoxy-5-nitro-, 2-hydroxy-5-nitro- and p-nitro-phenylalanine); furyl-, thienyl-, pyridyl-, pyrimidinyl-, purinyl- or naphthyl-alanines; and 20 tryptophan analogues and derivatives including kynurenine, 3- hydroxykynurenine, 2-hydroxytryptophan and 4-carboxytryptophan; α-Amino substituted amino acids including sarcosine (N-methylglycine), N-benzylglycine, N-methylalanine, N-benzylalanine, N-methylphenylalanine, N-benzylphenylalanme, N-methylvaline and N-benzylvaline; and 25 α-Hydroxy and substituted α -hydroxy amino acids including serine, threonine, allothreonine, phosphoserine and phosphothreonirte.

Polypeptides are polymers of amino acids in which a carboxyl group of one amino acid monomer is bonded to an amino or imino group of the next amino acid monomer by an amide bond. Polypeptides include dipeptides, low 30 molecular weight polypeptides (about 1500-5000 MW) and proteins. Proteins optionally contain 3, 5, 10, 50, 75, 100 or more residues, and suitably are substantially sequence-homologous with human, animal, plant or microbial

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5 proteins. They include enzymes (e.g., hydrogen peroxidase) as well as . immunogens such as KLH, or antibodies or proteins of any type against -which one wishes to raise an immune response. The nature and identity of the polypeptide may vary widely.

The polypeptide amidates are useful as immunogens in raising antibodies

10 against either the polypeptide (if it is not immunogenic in the animal to which it is administered) or against the epitopes on the remainder of the compound of this invention.

Antibodies capable of binding to the parental non-peptidyl compound are used to separate the parental compound from mixtures, for example in diagnosis

15 or manufacturing of the parental compound. The conjugates of parental compound and polypeptide generally are more immunogenic than the polypeptides in closely homologous animals, and therefore make the polypeptide more immunogenic for facilitating raising antibodies against it. Accordingly, the polypeptide or protein may be immunogenic in an animal

20 typically used to raise antibodies, e.g., rabbit, mouse, horse, or rat. The polypeptide optionally contains a peptidolytic enzyme cleavage site at the peptide bond between the first and second residues adjacent to the acidic heteroatom. Such cleavage sites are flanked by enzymatic recognition structures, e.g. a particular sequence of residues recognized by a peptidolytic enzyme.

25 Peptidolytic enzymes for cleaving the polypeptide conjugates of this invention are well known, and in particular include carboxypeptidases, which digest polypeptides by removing C-terminal residues, and are specific in many instances for particular C-terminal sequences. Such enzymes and their substrate requirements in general are well known. For example, a dipeptide (having a

30 given pair of residues and a free carboxyl terminus) is covalently bonded through its α-amino group to the phosphorus or carbon atoms of the compounds herein. In certain embodiments, a phosphonate group substituted with an

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5 amino acid or peptide will be cleaved by the appropriate peptidolytic enzyme, leaving the carboxyl of the proximal amino acid residue to autocatalytically cleave the phosphonoamidate bond.

Suitable dipeptidyl groups (designated by their single letter code) are AA, AR, AN, AD, AC, AE, AQ, AG, AH, AI, AL, AK, AM, AF, AP, AS, AT, AW, AY,

10 AV, RA, RR, RN, RD, RC, RE, RQ, RG, RH, RI, RL, RK, RM, RF, RP, RS, RT, RW, RY, RV, NA, NR, NN, ND, NC, NE, NQ, NG, NH, NI, NL, NK, NM, NF, NP, NS, NT, NW, NY, NV, DA, DR, DN, DD, DC, DE, DQ, DG, DH, DI, DL, DK, DM, DF, DP, DS, DT, DW, DY, DV, CA, CR, CN, CD, CC, CE, CQ, CG, CH, CI, CL, CK, CM, CF, CP, CS, CT, CW, CY, CV, EA, ER, EN, ED, EC, EE, EQ, EG, EH, EI, EL,

15 EK, EM, EF, EP, ES, ET, EW, EY, EV, QA, QR, QN, QD, QC, QE, Qa QG, QH, QI, QL, QK, QM, QF, QP, QS, QT, QW, QY, QV, GA, GR, GN, GD, GC, GE, GQ, GG, GH, GI, GL, GK, GM, GF, GP, GS, GT, GW, GY, GV, HA, HR, HN, HD, HQ HE, Ha HG, HH, HI, HL, HK, HM, HF, HP, HS, HT, HW, HY, HV, IA, IR, IN, ID, IQ IE, ia IG, IH, DC IL, IK, IM, IF, IP, IS, IT, IW, IY, IV, LA, LR, LN, LD, LC, LE, La

20 LG, LH, LI, LL, LK, LM, LF, LP, LS, LT, LW, LY, LV, KA, KR, KN, KD, KC, KE, Ka KG, KH, KI, KL, KK, KM, KF, KP, KS, KT, KW, KY, KV, MA, MR, MN, MD, MC, ME, Ma MG, MH, MI, ML, MK, MM, MF, MP, MS, MT, MW, MY, MV, FA, FR, FN, FD, FC, FE, Fa FG, FH, FI, FL, FK, FM, FF, FP, FS, FT, FW, FY, FV, PA, PR, PN, PD, PC, PE, Pa ^G, PH, PI, PL, PK, PM, PF, PP, PS, PT, PW, PY, PV, SA,

25 SR, SN, SD, SC, SE, Sa SG, SH, SI, SL, SK, SM, SF, SP, SS, ST, SW, SY, SV, TA, TR, TN, TD, TC, TE, Ta TG, TH, TI, TL, TK, TM, TF, TP, TS, TT, TW, TY, TV, WA, WR, WN, WD, WC, WE, WQ, WG, WH, WI, WL, WK, WM, WF, WP, WS, WT, WW, WY, WV, YA, YR, YN, YD, YC, YE, Ya YG, YH, YI, YL, YK, YM, YF, YP, YS, YT, YW, YY, YV, VA, VR, VN, VD, VC, VE, Va VG, VH, VI, VL, VK,

30 VM, VF, VP, VS, VT, VW, VY and W.

Tripeptide residues are also useful as protecting groups. When a phosphonate is to be protected, the sequence -X ⁴⁰ -pro- ⁵⁰ - (where X ⁴⁰ is any amino

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5 add residue and X ⁵⁰ is an amino acid residue, a carboxyl ester of proline, or hydrogen) will be cleaved by luminal carboxypeptidase to yield X ⁴⁰ with a free carboxyl, which in turn is expected to autocatalytically cleave the phosphonoamidate bond. The carboxy group of X ⁵⁰ optionally is esterified with benzyl.

10 Dipeptide or tripeptide species can be selected on the basis of known transport properties and/or susceptibility to peptidases that can affect transport to intestinal mucosal or other cell types. Dipeptides and tripeptides lacking an α-amino group are transport substrates for the peptide transporter found in brush border membrane of intestinal mucosal cells (Bai, J.P.F., (1992) Pharm Res.

15 9:969-978. Transport competent peptides can thus be used to enhance bioavailability of the amidate compounds. Di- or tripeptides having one or more amino acids in the D configuration may be compatible with peptide transport. Amino acids in the D configuration can be used to reduce the susceptibility of a di- or tripeptide to hydrolysis by proteases common to the brush border such as

20 aminopeptidase N. In addition, di- or tripeptides alternatively are selected on the basis of their relative resistance to hydrolysis by proteases found in the lumen of the intestine. For example, tripeptides or polypeptides lacking asp and/or glu are poor substrates for aminopeptidase A, di- or tripeptides lacking amino acid residues on the N-terminal side of hydrophobic amino acids (leu, tyr,

25 phe, val, trp) are poor substrates for endopeptidase, and peptides lacking a pro residue at the penultimate position at a free carboxyl terminus are poor substrates for carboxypeptidase P. Similar considerations can also be applied to the selection of peptides that are either relatively resistant or relatively susceptible to hydrolysis by cytosolic, renal, hepatic, serum or other peptidases.

30 Such poorly cleaved polypeptide amidates are immunogens or are useful for bonding to proteins in order to prepare immunogens.

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5 Intracellular Targeting

The phosphonate group of Formula I-IV compounds may cleave in vivo in stages after they have reached the desired site of action, i.e. inside a cell. One mechanism of action inside a cell may entail a first cleavage, e.g. by esterase, to provide a negatively-charged "locked-in" intermediate. Cleavage of a terminal

10 ester grouping in Formula I-IV compounds thus affords an unstable intermediate which releases a negatively charged "locked in" intermediate.

After passage inside a cell, intracellular enzymatic cleavage or modification of the phosphonate prodrug compound may result in an intracellular accumulation or retention of the cleaved or modified compound by

15 a "trapping"' mechanism. The cleaved or modified compound, i.e. active metabolite, may then be "locked-in" the cell, i.e. accumulate in the cell by a significant change in charge, polarity, or other physical property change which decreases the rate at which the cleaved or modified compound can exit the cell, relative to the rate at which it entered as the phosphonate prodrug. Other

20 mechanisms by which a therapeutic effect is achieved may be operative as well. Enzymes which are capable of an enzymatic activation mechanism with the phosphonate prodrug compounds of the invention include, but are not limited to, amidases, esterases, microbial enzymes, phospholipases, cholinesterases, and phosphatases.

25 It is known that the drug is activated in vivo by phosphorylation. Such . activation may occur in the present system by enzymatic conversion of the "locked-in" intermediate with phosphokinase to the active phosphonate diphosphate and/or by phosphorylation of the drug itself after its release from the "locked-in" intermediate as described above. In either case, the original

30 nucleoside-type drug will be converted, via the derivatives of this invention, to the active phosphorylated species.

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5 From the foregoing, it will be apparent that many structurally different known approved and experimental HCV polymerase inhibitor drugs can be derivatized in accord with the present invention. Numerous such drugs are specifically mentioned herein. However, it should be understood that the discussion of drug families and their specific members for derivatization

10 according to this invention is not intended to be exhaustive, but merely illustrative.

As another example, when the selected drug contains multiple reactive hydroxyl functions, a mixture of intermediates and final products may again be obtained. In the unusual case in which all hydroxy groups are approximately

15 equally reactive, there is not expected to be a single, predominant product, as each mono-substituted product will be obtained in approximate by equal amounts, while a lesser amount of multiply-substituted product will also result. Generally speaking, however, one of the hydroxyl groups will be more susceptible to substitution than the other(s), e.g. a primary hydroxyl will be

20 more reactive than a secondary hydroxyl, an unhindered hydroxyl will be more reactive than a hindered one. Consequently, the major product will be a mono- substituted one in which the most reactive hydroxyl has been derivatized while other mono-substituted and multiply-substituted products may be obtained as minor products.

25 Stereoisomers

The compounds of the invention, exemplified by Formula I, II, III or IV may have chiral centers, e.g. 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

30 compounds of the invention include enriched or resolved optical isomers at any or all asymmetric, chiral atoms. In other words, the chiral centers apparent from

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5 the depictions are provided as the chiral isomers or racemic mixtures. Both racemic and diastereomeric mixtures, as well as the individual optical isomers isolated or synthesized, substantially free of their enantiomeric or diastereomeric partners, are all within the scope of the invention. The racemic mixtures are separated into their individual, substantially optically pure isomers through

10 well-known techniques such as, for example, the separation of diastereomeric salts formed with optically active adjuncts, e.g., acids or bases followed by conversion back to the optically active substances/ In most instances, the desired optical isomer is synthesized by means of stereospecific reactions, beginning with the appropriate stereoisomer of the desired starting material.

15 The compounds of the invention can also exist as tautomeric isomers in certain cases. Although only one delocalized resonance structure may be depicted, all such forms are contemplated within the scope of the invention. For example, ene-amine tautomers can exist for purine, pyrϊmidine, imidazole, guanidine, amidine, and tetrazole systems and all their possible tautomeric

20 forms are within the scope of the invention.

Metabolites of the Compounds of the Invention

Also falling within the scope of this invention are the in vivo metabolic products of the compounds described herein, to the extent such products are novel and unobvious over the prior art. Such products may result for example 25 from the oxidation, reduction, hydrolysis, amidation, esterification and the like of the administered compound, primarily due to enzymatic processes. Accordingly, the invention includes novel and unobvious compounds produced by a process comprising contacting a compound of this invention with a mammal for a period of time sufficient to yield a metabolic product thereof.

30 Such products typically are identified by preparing a radiolabeled (e.g. ^C or ^H) compound of the invention, administering it parenterally in a detectable

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5 dose (e.g. greater than about 0.5 mg/kg) to an animal such as rat, mouse, guinea pig, monkey, or to man, allowing sufficient time for metabolism to occur (typically about 30 seconds to 30 hours) and isolating its conversion products from the urine, blood or other biological samples. These products are easily isolated since they are labeled (others are isolated by the use of antibodies

10 capable of binding epitopes surviving in the metabolite). The metabolite structures are determined in conventional fashion, e.g. by MS or NMR analysis. In general, analysis of metabolites is done in the same way as conventional drug metabolism studies well-known to those skilled in the art. The conversion products, so long as they are not otherwise found in vivo, are useful in diagnostic

15 assays for therapeutic dosing of the compounds of the invention even if they possess no HCV polymerase inhibitory activity of their own.

Recipes and methods for determining stability of compounds in surrogate gastrointestinal secretions are known. Compounds are defined herein as stable in the gastrointestinal tract where less than about 50 mole percent of the

20 protected groups are deprotected in surrogate intestinal or gastric juice upon incubation for 1 hour at 37°C. Simply because the compounds are stable to the gastrointestinal tract does not mean that they cannot be hydrolyzed in vivo. The phosphonate prodrugs of the invention typically will be stable in the digestive system but may be substantially hydrolyzed to the parental drug in the digestive

25 lumen, liver or other metabolic organ, or within cells in general.

Exemplary Methods of Making the Compounds of the Invention.

The invention provides many methods of making the compositions of the invention. The compositions are prepared by any of the applicable techniques of organic synthesis. Many such techniques are well known in the art, such as 30 those elaborated in "Compendium of Organic Synthetic Methods" (John Wiley & Sons, New York), Vol. 1, Ian T. Harrison and Shuyen Harrison, 1971; Vol. 2, Ian

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5 T. Harrison and Shuyen Harrison, 1974; Vol. 3, Louis S. Hegedus and Leroy

Wade, 1977; Vol. 4, Leroy G. Wade, jr., 1980; Vol. 5, Leroy G. Wade, Jr., 1984; and Vol. 6, Michael B. Smith; as well as March, J., "Advanced Organic Chemistry, Third Edition", Qohn Wiley & Sons, New York, 1985), "Comprehensive Organic Synthesis. Selectivity, Strategy & Efficiency in Modern Organic Chemistry. In 9

10 Volumes", Barry M. Trost, Editor-in-Chief (Pergamon Press, New York, 1993 printing).

Dialkyl phosphonates may be prepared according to the methods of: Quast et al (1974) Synthesis 490; Stowell et al (1990) Tetrahedron Lett. 3261; US Patent No. 5,663,159.

15 In general, synthesis of phosphonate esters is achieved by coupling a nucleophile amine or alcohol with the corresponding activated phosphonate electrophilic precursor. For example, chlorophosphonate addition on to 5'- hydroxy of nucleoside is a well known method for preparation of nucleoside phosphate monoesters. The activated precursor can be prepared by several well

20 known methods. Chlorophosphonates useful for synthesis of the prodrugs are prepared from the substituted-l,3-propanediol (Wissner, et al, (1992) /. Med Chem. 35:1650). Chlorophosphonates are made by oxidation of the corresponding chlorophospholanes (Anderson, et al, (1984) /. Org. Chem. 49:1304) which are obtained by reaction of the substituted diol with phosphorus

25 trichloride. Alternatively, the chlorophosphonate agent is made by treating substituted-l _/ 3-diols with phosphorusoxychloride (Patois, et al, (1990) /. Chem. Soc. Perkin Trans. 1, 1577). Chlorophosphonate species may also be generated in situ from corresponding cyclic phosphites (Silverburg, et al., (1996) Tetrahedron Lett, 37:771-774), which in turn can be either made from chlorophospholane or

30 phosphoramidate intermediate. Phosphoroflouridate intermediate prepared either from pyrophosphate or phosphoric acid may also act as precursor in

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5 preparation of cyclic prodrugs (Watanabe et al., (1988) Tetrahedron Lett, 29:5763- 66). Caution: fluorophosphonate compounds may be highly toxic!

Phosphonate prodrugs of the present invention may also be prepared from the precursor free acid by Mitsunobu reactions (Mitsunobu, (1981) Synthesis, 1; Campbell, (1992) /. Org. Chem., 52:6331), and other acid coupling

10 reagents including, but not limited to, carbodiimides (Alexander, et al, (1994) Collect Czech. Chem. Commun. 59:1853; Casara, et al, (1992) Bioorg. Med. Chem. Lett., 2:145; Ohashi, et al, (1988) Tetrahedron Lett., 29:1189), and benzotriazolyloxytris-(dimethylamino)phosphonium salts (Campagne, et al, (1993) Tetrahedron Lett, 34:6743).

15 Aryl halides undergo Ni ⁺² catalyzed reaction with phosphite derivatives to give aryl phosphonate containing compounds (Balthazar, et al (1980) J. Org. Chem. 45:5425). Phosphonates may also be prepared from the chlorophosphonate in. the presence of a palladium catalyst using aromatic triflates (Petrakis, et al, (1987) /. Am. Chem. Soc.109:2831; Lu, et al, (1987) Synthesis,

20 726). In another method, aryl phosphonate esters are prepared from aryl phosphates under anionic rearrangement conditions (Melvin (1981) Tetrahedron Lett. 22:3375; Casteel, et al, (1991) Synthesis, 691). N-Alkoxy aryl salts with alkali metal derivatives of cyclic alkyl phosphonate provide general synthesis for heteroaryl-2-phosphόnate linkers (Redmore (1970) /. Org. Chem. 35:4114), These

25 above mentioned methods can also be extended to compounds where the W ⁵ group is a heterocycle. Cyclic-l^-propanyl prodrugs of phosphonates are also synthesized from phosphonic diacids and substituted propane-l,3-diols using a coupling reagent such as 1,3-dicyclohexylcarbodiimide (DCC) in presence of a base (e.g., pyridine). Other carbodϋmide based coupling agents like 1,3-

30 disopropylcarbodiimide or water soluble reagent, l-(3-dimethylaminopropyl)-3- ethylcarbodiimide hydrochloride (EDCI) can also be utilized for the synthesis of cyclic phosphonate prodrugs.

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5 The carbamoyl group may be formed by reaction of a hydroxy group according to the methods known in the art, including the teachings of Ellis, US 2002/0103378 Al and Hajima, US Patent No.6,018,049.

Schemes and Examples

A number of exemplary methods for the preparation of the compositions

10 of the invention are provided below. These methods are intended to illustrate the nature of such preparations are not intended to limit the scope of applicable methods.

General aspects of these exemplary methods are described below and in the Examples. Each of the products of the following processes is optionally

15 separated, isolated, and/or purified prior to its use in subsequent processes. Generally, the reaction conditions such as temperature, reaction time, solvents, work-up procedures, and the like, will be those common in the art for the particular reaction to be performed. The cited reference material, together with material cited therein, contains detailed descriptions of such conditions.

20 Typically the temperatures will be -100 ⁰ C to 200 ⁰ C, solvents will be aprotic or protic, and reaction times will be 10 seconds to 10 days. Work-up typically consists of quenching any unreacted reagents followed by partition between a water/organic layer system (extraction) and separating the layer containing the product.

25 Oxidation and reduction reactions are typically carried out at temperatures near room temperature (about 20°C), although for metal hydride reductions frequently the temperature is reduced to 0 ⁰ C to -100 ⁰ C, solvents are typically aprotic for reductions and may be either protic or aprotic for oxidations. Reaction times are adjusted to achieve desired conversions.

30 Condensation reactions are typically carried out at temperatures near room temperature, although for non-equilibrating, kinetically controlled

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5 condensations reduced temperatures (0 ⁰ C to -100 ⁰ C) are also common. Solvents can be either protic (common in equilibrating reactions) or aprotic (common in kinetically controlled reactions).

Standard synthetic techniques such as azeotropic removal of reaction byproducts and use of anhydrous reaction conditions (e.g. inert gas environments)

10 are common in the art and will be applied when applicable.

The terms "treated", "treating", "treatment", and the like, mean contacting, mixing, reacting, allowing to react, bringing into contact, and other terms common in the art for indicating that one or more chemical entities is treated in such a manner as to convert it to one or more other chemical entities. This

15 means that "treating compound one with compound two" is synonymous with "allowing compound one to react with compound two", "contacting compound one with compound two", "reacting compound one with compound two", and other expressions common in the art of organic synthesis for reasonably indicating that compound one was "treated", "reacted", "allowed to react", etc.,

20 with compound two.

"Treating" indicates the reasonable and usual manner in which organic chemicals are allowed to react. Normal concentrations (0.01M to 1OM, typically 0.1M to IM), temperatures (-100 ⁰ C to 250 ⁰ C, typically -78°C to 150 ⁰ C, more typically -78°C to 100 ⁰ C, still more typically 0 ⁰ C to 100 ⁰ C), reaction vessels

25 (typically glass, plastic, metal), solvents, pressures, atmospheres (typically air for oxygen and water insensitive reactions or nitrogen or argon for oxygen or water sensitive), etc., are intended unless otherwise indicated. The knowledge of similar reactions known in the art of organic synthesis are used in selecting the conditions and apparatus for "treating" in a given process. In particular, one of

30 ordinary skill in the art of organic synthesis selects conditions and apparatus reasonably expected to successfully carry out the chemical reactions of the described processes based on the knowledge in the art.

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5 Modifications of each of the exemplary schemes above and in the examples (hereafter "exemplary schemes") leads to various analogs of the specific exemplary materials produce. The above cited citations describing suitable methods of organic synthesis are applicable to such modifications. In each of the exemplary schemes it may be advantageous to separate

10 reaction products from one another and/or from starting materials. The desired products of each step or series of steps is separated and/or purified (hereinafter separated) to the desired degree of homogeneity by the techniques common in the art. Typically such separations involve multiphase extraction, crystallization from a solvent or solvent mixture, distillation, sublimation, or chromatography.

15 Chromatography can involve any number of methods including, for example: reverse-phase and normal phase; size exclusion; ion exchange; high, medium, and low pressure liquid chromatography methods and apparatus; small scale analytical; simulated moving bed (SMB) and preparative thin or thick layer chromatography, as well as techniques of small scale thin layer and flash

20 chromatography.

Another class of separation methods involves treatment of a mixture with a reagent selected to bind to or render otherwise separable a desired product, unreacted starting material, reaction by product, or the like. Such reagents include adsorbents or absorbents such as activated carbon, molecular sieves, ion

25 exchange media, or the like. Alternatively, the reagents can be acids in the case of a basic material, bases in the case of an acidic material, binding reagents such as antibodies, binding proteins, selective chelators such as crown ethers, liquid/liquid ion extraction reagents (LIX), or the like.

Selection of appropriate methods of separation depends on the nature of

30 the materials involved. For example, boiling point, and molecular weight in distillation and sublimation, presence or absence of polar functional groups in chromatography, stability of materials in acidic and basic media in multiphase

133

5 extraction, and the like. One skilled in the art will apply techniques most likely to achieve the desired separation.

A single stereoisomer, e.g. an enantiomer, substantially free of its stereoisomer may be obtained by resolution of the racemic mixture using a method such as formation of diastereomers using optically active resolving

10 agents ("Stereochemistry of Carbon Compounds," (1962) by E. L. Eliel, McGraw Hill; Lochmuller, C. H., (1975) /. Chromatogr., 113:(3) 283-302). Racemic mixtures of chiral compounds of the invention can be separated and isolated by any suitable method, including: (1) formation of ionic, diastereomeric salts with chiral compounds and separation by fractional crystallization or other methods,

15 (2) formation of diastereomeric compounds with chiral derivatizing reagents, separation of the diastereomers, and conversion to the pure stereoisomers, and (3) separation of the substantially pure or enriched stereoisomers directly under chiral conditions.

Under method (1), diastereomeric salts can be formed by reaction of

20 enantiomerically pure chiral bases such as brucine, quinine, ephedrine, strychnine, α-methyl-β-phenylethylamine (amphetamine), and the like with asymmetric compounds bearing acidic functionality, such as carboxylic acid and sulfonic acid. The diastereomeric salts may be induced to separate by fractional crystallization or ionic chromatography. For separation of the optical isomers of

25 amino compounds, addition of chiral carboxylic or sulfonic acids, such as camphorsulf onic acid, tartaric acid, mandelic acid, or lactic acid can result in formation of the diastereomeric salts.

Alternatively, by method (2), the substrate to be resolved is reacted with one enantiomer of a chiral compound to form a diastereomeric pair (Eliel, E. and

30 Wilen, S. (1994) Stereochemistry of Organic Compounds, John Wiley & Sons, Inc., p. 322). Diastereomeric compounds can be formed by reacting asymmetric compounds with enantiomerically pure chiral derivatizing reagents, such as

134

5 menthyl derivatives, followed by separation of the diastereomers and hydrolysis to yield the free, enantiomerically enriched xanthene. A method of determining optical purity involves making chiral esters, such as a menthyl ester, e.g. (-) menthyl chloroformate in the presence of base, or Mosher ester, α-methoxy-α- (trifluoromethyl)phenyl acetate (Jacob III. (1982) /. Org. Chem. 47:4165), of the

10 racemic mixture, and analyzing the NMR spectrum for the presence of the two atropisomeric diastereomers. Stable diastereomers of atropisomeric compounds can be separated and isolated by normal- and reverse-phase chromatography following methods for separation of atropisomeric naphthyl-isoquinolines (Hoye, T., WO 96/15111). By method (3), a racemic mixture of two enantiomers can be

15 separated by chromatography using a chiral stationary phase ( Chiral Liquid Chromatography (1989) W. J. Lough, Ed. Chapman and Hall, New York; Okamoto, (1990) /. ofChromatogr. 513:375-378). Enriched or purified enantiomers can be distinguished by methods used to distinguish other chiral molecules with asymmetric carbon atoms, such as optical rotation and circular dichroism.

20 All literature and patent citations above are hereby expressly incorporated by reference at the locations of their citation. Specifically cited sections or pages of the above cited works are incorporated by reference with specificity. The invention has been described in detail sufficient to allow one of ordinary skill in the art to make and use the subject matter of the following

25 Embodiments. It is apparent that certain modifications of the methods and compositions of the following Embodiments can be made within the scope and spirit of the invention.

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Scheme A

O O R-link — P-OR ¹ R-link R-OR ¹

OR ¹ OH

27.1 27.2

O O

R-Ijnk — R-OR ¹ R-link — P-OH

OR ¹ OH

27.1 27.3

O O R-link — R-OR ¹ R-link — R-OH

OH OH 27.2 27.3

O 4 O R-link — P-OR ¹ R-link — R-OR ¹

OH W 27.2 27.1

O R-link — R-OH R-link — P-OR ¹

OH OH 27.3 27.2 p , O

R-link — R-OH R-link — R-OR ¹

OH OR ' 27.3 27.1

Scheme A shows the general interconversions of certain phosphonate compounds: acids -P(O)(OH)2; mono-esters -P(O)(ORi)(OH); and diesters -

10 P(O)(ORi)2 in which the R ¹ groups are independently selected, and defined herein before, and the phosphorus is attached through a carbon moiety (link, i.e. linker), which is attached to the rest of the molecule, e.g. drug or drug intermediate (R). The R ¹ . groups attached to the phosphonate esters in Scheme 1 may be changed using established chemical transformations. The

15 interconversions may be carried out in the precursor compounds or the final products using the methods described below. The methods employed for a given phosphonate transformation depend on the nature of the substituent R ¹ .

136

5 The preparation and hydrolysis of phosphonate esters is described in Organic Phosphorus Compounds, G. M. Kosolapoff, L. Maeir, eds, Wiley, 1976, p. 9ff.

The conversion of a phosphonate diester 27.1 into the corresponding phosphonate monoester 27.2 (Scheme A, Reaction 1) can be accomplished by a number of methods. For example, the ester 27.1 in which R ¹ is an arylalkyl

10 group such as benzyl, can be converted into the monoester compound 27.2 by reaction with a tertiary organic base such as diazabicyclooctane (DABCO) or quinuclidine, as described in/. Org. Chem., 1995, 60:2946. The reaction is performed in an inert hydrocarbon solvent such as toluene or xylene, at about 110 ⁰ C. The conversion of the diester 27.1 in which RHs an aryl group such as

15 phenyl, or an alkenyl group such as allyl, into the monoester 27.2 can be effected by treatment of the ester 27.1 with a base such as aqueous sodium hydroxide in acetonitrile or lithium hydroxide in aqueous tetrahydrofuran. Phosphonate diesters 27.2 in which one of the groups R ¹ is arylalkyl, such as benzyl, and the other is alkyl, can be converted into the monoesters 27.2 in which R ¹ is alkyl, by

20 hydrogenation, for example using a palladium on carbon catalyst. Phosphonate diesters in which both of the groups R ¹ are alkenyl, such as allyl, can be converted into the monoester 27.2 in which R ¹ is alkenyl, by treatment with chloro tris(triphenylphosphine)rhodium (Wilkinson's catalyst) in aqueous ethanol at reflux, optionally in the presence of diazabicyclooctane, for example

25 by using the procedure described in /. Org. Chem., 38:3224 1973 for the cleavage of allyl carboxylates.

The conversion of a phosphonate diester 27.1 or a phosphonate monoester 27.2 into the corresponding phosphonic acid 27.3 (Scheme A, Reactions 2 and 3) can effected by reaction of the diester or the monoester with trimethylsilyl

30 bromide, as described in /. Chem. Soc, Chem. Comm., 739, 1979. The reaction is conducted in an inert solvent such as, for example, dichloromethane, optionally in the presence of a silylating agent such as bis(trimethylsilyl)trifluoroacetamide,

137

S at ambient temperature. A phosphonate monoester 27.2 in which R ^a is arylalkyl such as benzyl, can be converted into the corresponding phosphonic add 27.3 by hydrogenation over a palladium catalyst, or by treatment with hydrogen chloride in an ethereal solvent such as dioxane. A phosphonate monoester 27.2 in which R ¹ is alkenyl such as, for example, allyl, can be converted into the

10 phosphonic acid 27.3 by reaction with Wilkinson's catalyst in an aqueous organic solvent, for example in 15% aqueous acetonitrile, or in aqueous ethanol, for example using the procedure described in HeIv. Chim. Acta., 68:618, 1985. Palladium catalyzed hydrogenolysis of phosphonate esters 27.1 in which R ¹ is benzyl is described in /. Org. Chern., 24:434, 1959. Platinum-catalyzed

15 hydrogenolysis of phosphonate esters 27.1 in which R ¹ is phenyl is described in /. Amer. Chem. Soc, 78:2336, 1956.

The conversion of a phosphonate monoester 27.2 into a phosphonate diester 27.1 (Scheme A, Reaction 4) in which the newly introduced R ¹ group is alkyl, arylalkyl, or haloalkyl such as chloroethyl, can be effected by a number of

20 reactions in which the substrate 27.2 is reacted with a hydroxy compound R ¹ OH, in the presence of a coupling agent. Suitable coupling agents are those employed for the preparation of carboxylate esters, and include a carbodiimide such as dicyclohexylcarbodiimide, in which case the reaction is preferably conducted in a basic organic solvent such as pyridine, or (benzotriazol-1-

25 yloxy)tripyrrόlidinophosphonium hexafluorophosphate (PYBOP, Sigma), in which case the reaction is performed in a polar solvent such as dimethylformamide, in the presence of a tertiary organic base such as diisopropylethylamine, or Aldrithiol-2 (Aldrich) in which case the reaction is conducted in a basic solvent such as pyridine, in the presence of a triaryl

30 phosphine such as triphenylphosphine. Alternatively, the conversion of the phosphonate monoester 27.1 to the diester 27.1 can be effected by the use of the . Mitsunobu reaction. The substrate is reacted with the hydroxy compound R ¹ OH,

138

5 in the presence of diethyl azodicarboxylate and a triarylphosphine such as triphenyl phosphine. Alternatively, the phosphonate monoester 27.2 can be transformed into the phosphonate diester 27.1, in which the introduced R ¹ group is alkenyl or arylalkyl, by reaction of the monoester with the halide R ¹ Br, in which R ¹ is as alkenyl or arylalkyl. The alkylation reaction is conducted in a

10 polar organic solvent such as dimethylformarrd.de or acetonitrile, in the presence of a base such as cesium carbonate. Alternatively, the phosphonate monoester can be transformed into the phosphonate diester in a two step procedure. In the first step, the phosphonate monoester 27.2 is transformed into the chloro analog -P(O)(OR ¹ )C1 by reaction with thionyl chloride or oxalyl chloride and the like, as

15 described in Organic Phosphorus Compounds, G. M. Kosolapoff, L. Maeir, eds, Wiley, 1976, p. 17, and the thus-obtained product -P(O)(OR ¹ )C1 is then reacted with the hydroxy compound R ¹ OH, in the presence of a base such as triethylamine, to afford the phosphonate diester 27.1.

A phosphonic acid -P(O)(OH>2 can be transformed into a phosphonate

20 monoester -P(O)(OR ¹ XOH) (Scheme A, Reaction 5) by means of the methods described above of for the preparation of the phosphonate diester -P(O)(OR ¹ )2 27.1, except that only one molar proportion of the component R ¹ OH or R ¹ Br is employed.

A phosphonic acid -P(O)(OH)227.3 can be transformed into a

25 phosphonate diester -P(O)(OR ¹ )227.1 (Scheme A, Reaction 6) by a coupling reaction with the hydroxy compound R ¹ OH, in the presence of a coupling agent such as Aldrithiol-2 (Aldrich) and triphenylphosphine. The reaction is conducted in a basic solvent such as pyridine. Alternatively, phosphonic acids 27.3 can be transformed into phosphonic esters 27.1 in which R ¹ is aryl, such as

30 phenyl, by means of a coupling reaction employing, for example, phenol and dicyclohexylcarbodiimide in pyridine at about 7O ⁰ C. Alternatively, phosphonic acids 27.3 can be transformed into phosphonic esters 27.1 in which R ¹ is alkenyl,

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5 by means of an alkylation reaction. The phosphonic acid is reacted with the alkenyl bromide R ¹ Br in a polar organic solvent such as acetonitrile solution at reflux temperature, in the presence of a base such as cesium carbonate, to afford the phosphonic ester 27.1.

Phosphonate prodrugs of the present invention may also be prepared

10 from the precursor free acid by Mitsunobu reactions (Mitsunobu, (1981)

Synthesis, 1; Campbell, (1992) /. Org. Chem., 52:6331), and other acid coupling reagents including, but not limited to, carbodiimides (Alexander, et al, (1994) Collect. Czech. Chem. Comtnun. 59:1853; Casara, et al, (1992) Bioorg. Med. Chem. Lett., 2:145; Ohashi, et al, (1988) Tetrahedron Lett., 29:1189), and

15 benzotriazolyloxytris-(dimethylamino)phosphonium salts (Campagne, et al, (1993) Tetrahedron Lett., 34:6743).

Preparation of carboalkoxy-substituted phosphonate bisamidates, monoamidates, diesters and monoesters. 20

A number of methods are available for the conversion of phosphonic acids into amidates and esters. In one group of methods, the phosphonic acid is either converted into an isolated activated intermediate such as a phosphoryl chloride, or the phosphonic acid is activated in situ for reaction with an amine or 25 a hydroxy compound.

The conversion of phosphonic acids into phosphoryl chlorides is accomplished by reaction with thionyl chloride, for example as described in J.

Gen. Chem. USSR, 1983, 53, 480, Zh. Obschei Khim., 1958, 28, 1063, or J. Org.

Chem., 1994, 59, 6144, or by reaction with oxalyl chloride, as described in J. Am. 30 Chem. Soc, 1994, 116, 3251, or J. Org. Chem., 1994, 59, 6144, or by reaction with phosphorus pentachloride, as described in J. Org. Chem., 2001, 66, 329, or in J.

Med. Chem., 1995, 38, 1372. The resultant phosphoryl chlorides are then reacted

140

5 with amines or hydroxy compounds in the presence of a base to afford the amidate or ester products.

Phosphonic acids are converted into activated imidazolyl derivatives by reaction with carbonyl diimidazole, as described in J. Chem. Soα, Chem. Comm., 1991, 312, or Nucleosides Nucleotides 2000, 19, 1885. Activated sulfonyloxy

10 derivatives are obtained by the reaction of phosphonic acids with trichloromethylsulfonyl chloride, as described in J. Med. Chem. 1995, 38, 4958, or with triisopropylbenzenesulfonyl chloride, as described in Tet. Lett., 1996, 7857, or Bioorg. Med. Chem. Lett., 1998, 8, 663. The activated sulfonyloxy derivatives are then reacted with amines or hydroxy compounds to afford amidates or esters.

15 Alternatively, the phosphonic acid and the amine or hydroxy reactant are combined in the presence of a diimide coupling agent. The preparation of phosphonic amidates and esters by means of coupling reactions in the presence of dicyclohexyl carbodiimide is described, for example, in J. Chem. Soc, Chem. Comm, 1991, 312, or J. Med. Chem., 1980, 23, 1299 or Coll. Czech. Chem. Comm.,

20 1987, 52, 2792. The use of ethyl dimethylaminopropyl carbodiimide for activation and coupling of phosphonic acids is described in Tet. Lett., 2001, 42, 8841, or Nucleosides Nucleotides, 2000, 19, 1885.

A number of additional coupling reagents have been described for the preparation of amidates and esters from phosphonic acids. The agents include

25 Aldrithiol-2, and PYBOP and BOP, as described in J. Org. Chem., 1995, 60, 5214, and J. Med. Chem., 1997, 40, 3842, mesitylene-2-sulfonyl-3-nitro-l,2,4-triazole (MSNT), as described in J. Med. Chem., 1996, 39, 4958, diphenylphosphoryl azide, as described in J. Org. Chem., 1984, 49, 1158, l-(2,4,6- triisopropylbenzenesulfonyl-3-nitro-l,2,4-triazole (TPSNT) as described in

30 Bioorg. Med. Chem. Lett., 1998, 8, 1013, bromotris(dimethylamino)phosphonium hexailuorophosphate (BroP), as described in Tet. Lett., 1996, 37, 3997, 2-chloro- 5,5-dimethyl-2-oxo-l,3,2-dioxaphosphinane, as described in Nucleosides

141

5 Nucleotides 1995, 14, 871, and diphenyl chlorophosphate, as described in J. Med. Chem., 1988, 31, 1305.

Thosphonic acids are converted into amidates and esters by means of the Mitsonobu reaction, in which the phosphonic acid and the amine or hydroxy reactant are combined in the presence of a triaryl phosphine and a dialkyl 10 azodicarboxylate. The procedure is described in Org. Lett., 2001, 3, 643, or J. Med. Chem., 1997, 40, 3842.

Phosphonic esters are also obtained by the reaction between phosphonic acids and halo compounds, in the presence of a suitable base. The method is described, for example, in Anal. Chem., 1987, 59, 1056, or J. Chem. Soc. Perkin 15 Trans., 1, 1993, 19, 2303, or J. Med. Chem., 1995, 38, 1372, or Tet. Lett., 2002, 43, 1161.

Schemes 1 - 4 illustrate the conversion of phosphonate esters and phosphonic acids into carboalkoxy-substituted phosphorobisamidates (Scheme T), phosphoroamidates (Scheme 2), phosphonate monoesters (Scheme 3) and 20 phosphonate diesters, (Scheme 4).

Scheme 1 illustrates various methods for the conversion of phosphonate diesters 1.1 into phosphorobisamidates 1.5. The diester 1.1, prepared as described previously, is hydrolyzed, either to the monoester 1.2 or to the phosphonic acid 1.6. The methods employed for these transformations are 25 described above. The monoester 1.2 is converted into the monoamddate 1.3 by reaction with an amirioester 1.9, in which the group R ² is H or aϊkyl, the group R ⁴ is an alkylene moiety such as, for example, CHCH ₃ , CHPr ¹ , CH(CH-Ph), CHiCH(CHs) and the like, or a group present in natural or modified aminoacids, and the group R ⁵ is alkyl. The reactants are combined in the presence of a 30 coupling agent such as a carbodiimide, for example dicyclohexyl carbodiimide, as described in J. Am. Chem. Soc, 1957, 79, 3575, optionally in the presence of an activating agent such as hydroxybenztriazole, to yield the amidate product 1.3.

142

5 The amidate-forming reaction is also effected in the presence of coupling agents such as BOP, as described in J. Org. Chem., 1995, 60, 5214, Aldrithiol, PYBOP and similar coupling agents used for the preparation of amides and esters. Alternatively, the reactants 1.2 and 1.9 are transformed into the monoamidate 1.3 by means of a Mitsonobu reaction. The preparation of amidates by means of the

10 Mitsonobu reaction is described in J. Med. Chem., 1995, 38, 2742. Equimolar amounts of the reactants are combined in an inert solvent such as tetrahydrofuran in the presence of a triaryl phosphine and a dialkyl azodicarboxylate. The thus-obtained monoamidate ester 1.3 is then transformed into amidate phosphonic acid 1.4. The conditions used for the hydrolysis

15 reaction depend on the nature of the R ¹ group, as described previously. The phosphonic acid amidate 1.4 is then reacted with an aminoester 1.9, as described above, to yield the bisamidate product 1.5, in -which the amino substituents are the same or different.

An example of this procedure is shown in Scheme 1, Example 1. In this

20 procedure, a dibenzyl phosphonate 1.14 is reacted with diazabicyclooctane (DABCO) in toluene at reflux, as described in J. Org. Chem., 1995, 60, 2946, to afford the monobenzyl phosphonate 1.15. The product is then reacted with equimolar amounts of ethyl alaninate 1.16 and dicyclohexyl carbodiimide in pyridine, to yield the amidate product 1.17. The benzyl group is then removed,

25 for example by hydrogenolysis over a palladium catalyst, to give the monoacid product 1.18. This compound is then reacted in a Mitsonobu reaction with ethyl leucinate 1.19, triphenyl phosphine and diethylazodicarboxylate, as described in J. Med. Chem., 1995, 38, 2742, to produce the bisamidate product 1.20.

Using the above procedures, but employing, in place of ethyl leucinate

30 1.19 or ethyl alaninate 1.16, different aminoesters 1.9, the corresponding products 1.5 are obtained.

143

5 Alternatively, the phosphonic acid 1.6 is converted into the bisamidate 1.5 by use of the coupling reactions described above. The reaction is performed in one step, in which case the nitrogen-related substituents present in the product 1.5 are the same, or in two steps, in which case the nitrogen-related substituents can be different.

10 An example of the method is shown in Scheme 1, Example 2. In this procedure, a phosphonic acid 1.6 is reacted in pyridine solution with excess ethyl phenylalaninate 1.21 and dicyclohexylcarbodiimide, for example as described in J. Chem. Soo, Chem. Comm., 1991, 1063, to give the bisamidate product 1.22. Using the above procedures, but employing, in place of ethyl

15 phenylalaninate, different aminoesters 1.9, the corresponding products 1.5 are obtained.

As a further alternative, the phosphonic acid 1.6 is converted into the mono or bis-activated derivative 1.7, in which Lv is a leaving group such as chloro, imidazolyl, triisopropylbenzenesulfonyloxy etc. The conversion of

20 phosphonic acids into chlorides 1.7 (Lv = Cl) is effected by reaction with thionyl chloride or oxalyl chloride and the like, as described in Organic Phosphorus Compounds, G. M. Kosolapoff, L. Maeir, eds, Wiley, 1976, p. 17. The conversion of phosphonic acids into monoimidazolides 1.7 (Lv = imidazolyl) is described in J. Med. Chem., 2002, 45, 1284 and in J. Chem. Soc. Chem. Comm., 1991, 312.

25 Alternatively, the phosphonic acid is activated by reaction with triisopropylbenzenesulfonyl chloride, as described in Nucleosides and Nucleotides, 2000, 10, 1885. The activated product is then reacted with the aminoester 1.9, in the presence of a base, to give the bisamidate 1.5. The reaction is performed in one step, in which case the nitrogen substituents present in the

30 product 1.5 are the same, or in two steps, via the intermediate 1.11, in which case the nitrogen substituents can be different.

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5 Examples of these methods are shown in Scheme 1, Examples 3 and 5. In the procedure illustrated in Scheme 1, Example 3, a phosphonic acid 1.6 is reacted with ten molar equivalents of thionyl chloride, as described in Zh. Obschei Khim., 1958, 28, 1063, to give the dichloro compound 1.23. The product is then reacted at reflux temperature in a polar aprotic solvent such as

10 acetonitrile, and in the presence of a base such as triethylamine, with butyl serinate 1.24 to afford the bisamidate product 1.25.

Using the above procedures, but employing, in place of butyl serinate 1.24, different aminoesters 1.9, the corresponding products 1.5 are obtained.

In the procedure illustrated in Scheme 1, Example 5, the phosphonic acid

15 1.6 is reacted, as described in J. Chem. Soc. Chem. Comm., 1991, 312, with carbonyl diimidazole to give the imidazolide 1.32. The product is then reacted in acetonitrile solution at ambient temperature, with one molar equivalent of ethyl alaninate 1.33 to yield the monodisplacement product 1.34. The latter compound is then reacted with carbonyl diimidazole to produce the activated

20 intermediate 1.35, and the product is then reacted, under the same conditions, with ethyl N-methylalaninate 1.33a to give the bisamidate product 1.36.

Using the above procedures, but employing, in place of ethyl alaninate 1.33 or ethyl N-methylalaninate 1.33a, different aminoesters 1.9, the corresponding products 1.5 are obtained.

25 The intermediate monoamidate 1.3 is also prepared from the monoester

1.2 by first converting the monoester into the activated derivative 1.8 in which Lv is a leaving group such as halo, imidazolyl etc, using the procedures described above. The product 1.8 is then reacted with an aminoester 1.9 in the presence of a base such as pyridine, to give an intermediate monoamidate

30 product 1.3. The latter compound is then converted, by removal of the R ¹ group and coupling of the product with the aminoester 1.9, as described above, into the bisamidate 1.5.

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5 An example of this procedure, in which the phosphonic acid is activated by conversion to the chloro derivative 1-26, is shown in Scheme 1, Example 4. In this procedure, the phosphonic monobenzyl ester 1.15 is reacted, in dichloromethane, with thionyl chloride, as described in Tet. Let., 1994, 35, 4097, to afford the phosphoryl chloride 1.26. The product is then reacted in

10 acetonitrile solution at ambient temperature with one molar equivalent of ethyl 3-amino-2-methylpropionate 1.27 to yield the monoamidate product 1.28. The latter compound is hydrogenated in ethyl acetate over a 5% palladium on carbon catalyst to produce the monoacid product 1.29. The product is subjected to a Mitsonobu coupling procedure, with equimolar amounts of butyl alaninate 1.30,

15 triphenyl phosphine, diethylazodicarboxylate and triethylamine in tetrahydrofuran, to give the bisamidate product 1.31.

Using the above procedures, but employing, in place of ethyl 3-amino-2- methylpropionate 1.27 or butyl alaninate 1.30, different aminoesters 1.9, the corresponding products 1.5 are obtained.

20 The activated phosphonic acid derivative 1.7 is also converted into the bisamidate 1.5 via the diamino compound 1.10. The conversion of activated phosphonic acid derivatives such as phosphoryl chlorides into the corresponding amino analogs 1.10, by reaction with ammonia, is described in Organic Phosphorus Compounds,, G. M. Kosolapoff, L. Maeir, eds, Wiley, 1976.

25 The diamino compound 1.10 is then reacted at elevated temperature with a haloester 1.12, in a polar organic solvent such as dimethylformamide, in the presence of a base such as dimethylaminopyridine or potassium carbonate, to yield the bisamidate 1.5.

An example of this procedure is shown in Scheme 1, Example 6. In this

30 method, a dichlorophosphonate 1.23 is reacted with ammonia to afford the diamide 1.37. The reaction is performed in aqueous, aqueous alcoholic or alcoholic solution, at reflux temperature. The resulting diamino compound is

146

5 then reacted with two molar equivalents of ethyl 2-bromo-3-methylbutyrate 1.38, in a polar organic solvent such as N-methylpyrrolidinone at ca. 150°C, in the presence of a base such as potassium carbonate, and optionally in the presence of a catalytic amount of potassium iodide, to afford the bisamidate product 1.39. Using the above procedures, but employing, in place of ethyl 2-bromo-3-

10 methylbutyrate 1.38, different haloesters 1.12 the corresponding products 1.5 are obtained.

The procedures shown in Scheme 1 are also applicable to the preparation of bisamidates in which the aminoester moiety incorporates different functional groups. Scheme 1, Example 7 illustrates the preparation of bisamidates derived

15 from tyrosine. In this procedure, the monoimidazolide 1.32 is reacted with propyl tyrosinate 1.40, as described in Example 5, to yield the monoamidate 1.41. The product is reacted with carbonyl diimidazole to give the imidazolide 1.42, and this material is reacted with a further molar equivalent of propyl tyrosinate to produce the bisamidate product 1.43.

20 Using the above procedures, but employing, in place of propyl tyrosinate

1.40, different aminoesters 1.9, the corresponding products 1.5 are obtained. The aminoesters employed in the two stages of the above procedure can be the same or different, so that bisamidates with the same or different amino substituents are prepared.

25 Scheme 2 illustrates methods for the preparation of phosphonate monoamidates.

In one procedure, a phosphonate monoester 1.1 is converted, as described in Scheme 1, into the activated derivative 1.8. This compound is then reacted, as described above, with an aminoester 1.9, in the presence of a base, to afford the

30 monoamidate product 2.1. The procedure is illustrated in Scheme 2, Example 1. In this method, a monophenyl phosphonate 2.7 is reacted with, for example, thionyl chloride, as described in J. Gen. Chem. USSR., 1983, 32, 367, to give the

147

5 chloro product 2.8. The product is then reacted, as described in Scheme 1, with ethyl alaninate 2.9, to yield the amidate 2.10.

Using the above procedures, but employing, in place of ethyl alaninate 2.9, different aminoesters 1.9, the corresponding products 2.1 are obtained.

Alternatively, the phosphonate monoester 1.1 is coupled, as described in 10 Scheme 1, with an aminoester 1.9 to produce the amidate 2.1. If necessary, the R ¹ substituent is then altered, by initial cleavage to afford the phosphonic acid 2.2. The procedures for this transformation depend on the nature of the R ¹ group, and are described above. The phosphonic acid is then transformed into the ester amidate product 2.3, by reaction with the hydroxy compound R ³ OH, in which 15 the group R ³ is aryl, heteroaryl, alkyl, cydoalkyl, haloalkyl etc, using the same coupling procedures (carbodiimide, Aldrithiol-2, PYBOP, Mitsonobu reaction etc) described in Scheme 1 for the coupling of amines and phosphonic acids.

148

Scheme 1

p Ha(R ⁴ )CO ₂ R ⁵ 1.12 ,0

R-link — R-N R-link — R"-LV _{λ q} R-link — F£ P-fN ²

\~™"H _A -* — H R-link- -Fa-NH ₂ \ / _R 4)

.NKR ⁴ JCO ₂ R^x ₆ (Lv or OH)

NH ₂ (Lv or OH) > '

(R ⁴ ) 1.10 1.7 1.11 CO ₂ R ⁵

CO ₂ R ⁰

1.9

1.5 o R ² p o .O

R-link— F(-N'^ ₄ ,CO ₂ R ⁵

R-link — fi-OR ¹ R-link — f(-OR ¹ - R-link — F(-OH- OH OH N , ^l ' OR ¹ (R ⁴ ) R ² 1.1 1.6 ^N CO ₂ R ⁵

1.9 O

R-link— F(-OR ¹ *-_ R-link — R^-OR ¹ R-link — F(-OH Lv R ² NH(R ⁴ JCO ₂ R ⁵ - ^N ~ ^R2 JM-R ²

1.9 (R ⁴ ) (R ⁴ )

1.8 CO ₂ R ⁵ ^CO ₂ R ⁵

1.3 1.4

Scheme 1 Example 1

H ₂ NGH(Me)CO ₂ Et p R-link — R-OBn R-llnk— rf°OH ^{1 16} _> R-link— F(°NW ^M ^—^ R-link— f£N^x ^Me

OBn OBn OB Inn A ' OH V

COOEt COOEt 1.14 1.15 1.17 1.18

PH

H ₂ NCH(CH ₂ Pr ^1- )CO ₂ Et R-liπk -FfN ,Me

NH -^

1.19 COOEt

Pr ¹ H ₂ C- (

COOEt 1.20

Scheme 1 Example 2

_o H ₂ NCH(Bn)CO ₂ Et _Q )-cθOEt

R-link — pfoH — ¹ - ²¹ », R-link — F(-NH OH NH

Bn- {

COOEt

¹ - ⁶ 1.22

149

Scheme 1 Example 3 OH

_Q H ₂ NCH(CH ₂ OH)CO ₂ Bu _o V-CO ₂ Bu

R-link — F(-OH R-link -If-Cl l^l^ R.iink—^NVl OH Cl NH

1.6 1.23 HO CO ₂ Bu

1.25

Scheme 1 Example 4

H ₂ NCH ₂ CH(Me)CO ₂ Et _λ

O O -j 27 P

R-link - -R-OBn ^• R-link — F(-OBπ *- R-link — R ₁ -OBn R-link — F(-OH OH Cl NH NH

1.15 1.26 V-CO ₂ Et

Me Me Me

H ₂ NCH(Me)CO ₂ Bu o y — Co ₂ Bu V CO ₂ Et

1.28 1.29 »►- R-link — F(-NH

1.30 NH

CO ₂ Et

Me

1.31

Scheme 1 Example 5

H ₂ NCH(Me)CO ₂ Et Me _N

O P V-CO ₂ Et R-link — f(-OH R-link — R-OH //

R-link — F^-NH

OH Im 1.33 OH 1.6 1.32 1.34

Me, Me

O V-CO ₂ Et

MeNHCH(Me)CO ₂ Et O V-CO ₂ Et R-link — F^-NH *- R-link — P _x -NH Im 1.33a N-Me

1.35 Me— (

CO ₂ Et 1.36

Scheme 1 Example 6

^Pr '\

O P BrCH(Pr ¹ JCO ₂ Et P /- ^co 2Et

R-link — R-Cl -*~ R-link — R-NH ₂ »- R-link — R^-NH Cl NH ₂ ¹ - ³⁸ NH

1.23 1.37 P* ⁴ — (

CO ₂ Et 1.39

150

Scheme 1 Example 7

HOv

R-link- -F^-OH R-link — R^-Im NH NH

_/ P H ₂ N ^' CO ₂ Pr R-!ink — R-OH

Im 1.40 /= ^* CO ₂ Pr

1.32 HO

1.41 1.42 HO

1.43

Examples of this method are shown in Scheme 2, Examples and 2 and 3. In the sequence shown in Example 2, a monobenzyl phosphonate 2.11 is transformed by reaction with ethyl alaninate, using one of the methods

10 described above, into the monoamidate 2.12. The benzyl group is then removed by catalytic hydrogenation in ethyl acetate solution over a 5% palladium on carbon catalyst, to afford the phosphonic acid amidate 2.13. The product is then reacted in dichloromethane solution at ambient temperature with equimolar amounts of l-(dimethylammopropyl)-3-ethylcarbodiimide and trifluoroethanol

15 2.14, for example as described in Tet. Lett., 2001, 42, 8841, to yield the amidate ester 2.15.

In the sequence shown in Scheme 2, Example 3, the monoamidate 2.13 is coupled, in tetrahydrofuran solution at ambient temperature, with equimolar amounts of dicyclohexyl carbodiimide and 4-hydroxy-N-methylpiperidine 2.16,

20 to produce the amidate ester product 2.17.

Using the above procedures, but employing, in place of the ethyl alaninate product 2.12 different monoacids 2.2, and in place of trifluoroethanol 2.14 or 4-hydroxy-N-methylpiperidine 2.16, different hydroxy compounds R ³ OH, the corresponding products 2.3 are obtained.

25 Alternatively, the activated phosphonate ester 1.8 is reacted with ammonia to yield the amidate 2.4. The product is then reacted, as described in Scheme 1, with a haloester 2.5, in the presence of a base, to produce the amidate

151

5 product 2.6. If appropriate, the nature of the R ¹ group is changed, using the procedures described above, to give the product 2.3. The method is illustrated in Scheme 2, Example 4. In this sequence, the monophenyl phosphoryl chloride 2.18 is reacted, as described in Scheme 1, with ammonia, to yield the amino product 2.19. This material is then reacted in N-methylpyrrolidinone solution at

10 170°C with butyl 2-bromo-3-phenylpropionate 2.20 and potassium carbonate, to afford the amidate product 2.21.

Using these procedures, but employing, in place of butyl 2-bromo-3- phenylpropionate 2.20, different haloesters 2.5, the corresponding products 2.6 are obtained.

15 The monoamidate products 2.3 are also prepared from the doubly activated phosphonate derivatives 1.7. In this procedure, examples of which are described in Synlett., 1998, 1, 73, the intermediate 1.7 is reacted with a limited amount of the aminoester 1.9 to give the mono-displacement product 1.11. The latter compound is then reacted with the hydroxy compound R ³ OH in a polar

20 organic solvent such as dimethylformamide, in the presence of a base such as diisopropylethylamine, to yield the monoamidate ester 2.3.

The method is illustrated in Scheme 2, Example 5. In this method, the phosphoryl dichloride 2.22 is reacted in dichloromethane solution with one molar equivalent of ethyl N-methyl tyrosinate 2.23 and dimethylaminopyridine,

25 to generate the monoamidate 2.24. The product is then reacted with phenol 2.25 in dimethylformamide containing potassium carbonate, to yield the ester amidate product 2.26.

Using these procedures, but employing, in place of ethyl N-methyl tyrosinate 2.23 or phenol 2.25, the aminoesters 1.9 and/or the hydroxy

30 compounds R ³ OH, the corresponding products 2.3 are obtained.

152

Scheme 2

,u

R-link — Ff-Lv . P , ^R2

R-link — R-N Lv 1.9 Lv(R ⁴ )

1.7 CO ₂ R ⁵ 1.11

}R ³ OH

O ,O o

R-link — f(-OR ¹ R-link — F(-OR ¹ - R-link — pf-OH *-. R-link — f(-OR ³

1.1 O υπH 1.9 ,N-R ² YI-R ²

/ N-R ² (R ⁴ ') (R p4 ⁴ _\ ) (R ⁴ )

R ² NH(R ⁴ JCO ₂ R^. ^V CO ₂ R ⁵ CO ₂ R ⁵ bo ₂ R ⁵ 1.9 2.1 2.2 2.3

O p Ha(R ⁴ )CO ₂ R ⁵ p

R-link — f(-OR ¹ R-link— F(-0R ¹ ^ R-link— fC-OR ¹ Lv NH ₂ ² - ⁵ ^NH

1.8 (R ⁴ )

CO ₂ R ⁵ 2-4 2.6

Scheme 2 Example 1

/P _^ H ₂ NCH(Me)CO ₂ Et ,p

R-link — F^-OPh- -^R-link— Ff-OPh ^- U- ² R-||nk— RrOPh OH Cl 2.9 \ _H

2.7 Me— {

2.8

CO ₂ Et 2.10

Scheme 2 Example 2

P _/ P CF ₃ CH ₂ OH _/ P

R-link — p(-OBn R-link — F(-OBπ R-link— fi-OH — - — →- R-link— R-OCH ₂ CF ₃ OH NH NH - 2."14 NH

Me— ( Me— ( Me— (

CO ₂ Et CO ₂ Et CO ₂ Et

2.11 2.12 2.13 2.15

Scheme 2 Example 3

R-link — OH F(-OH R-link — f^-O—ζ N-Me

NH NH ^ ' Me-( Me' Me— (

CO ₂ Et 2.16 CO ₂ Et

2.13 2.17

153

Scheme 2 Example 4

_Q BrCH(Bn)CO ₂ Bu O

R-link — pf-OPh R-link — F(-OPh »- R-"nk — F(-OPh Cl NH ₂ ² - ²⁰ NH

Bn — (

CO ₂ Bu

2.18 2.19 2.21

Scheme 2 Example 5

HO

p

CO ₂ Et R-link — R^-Cl PhOH R-link — R;-O— ^ )>

R-link — pf'CI 2.25 Cl 2.23 γ \ _v ^N"Me HO-(Z >

CO ₂ Et CO ₂ Et

2.22 2.24 2.26

Scheme 3 illustrates methods for the preparation of carboalkoxy- substituted phosphonate diesters in which one of the ester groups incorporates a carboalkoxy substituent. 10 In one procedure, a phosphonate monoester 1.1, prepared as described above, is coupled, using one of the methods described above, with a hydroxyester 3.1, in which the groups R ⁴ and R ⁵ are as described in Scheme 1.

For example, equimolar amounts of the reactants are coupled in the presence of a carbodiimide such as dicyclohexyl carbodiimide, as described in Aust. J. 15 Chem., 1963, 609, optionally in the presence of dimethylaminopyridine, as described in Tet, 1999, 55, 12997. The reaction is conducted in an inert solvent at ambient temperature.

The procedure is illustrated in Scheme 3, Example 1. In this method, a monophenyl phosphonate 3.9 is coupled, in dichloromethane solution in the 20 presence of dicyclohexyl carbodiimide, with ethyl 3-hydroxy-2- methylpropionate 3.10 to yield the phosphonate mixed diester 3.11.

154

5 Using this procedure, but employing, in place of ethyl 3-hydroxy-2- methylpropionate 3.10, different hydroxyesters 3.1, the corresponding products 3.2 are obtained.

The conversion of a phosphonate monoester 1.1 into a mixed diester 3.2 is also accomplished by means of a Mitsonobu coupling reaction with the 10 hydroxyester 3.1, as described in Org. Lett., 2001, 643. In this method, the reactants 1.1 and 3.1 are combined in a polar solvent such as tetrahydrofuran, in the presence of a triarylphosphine and a dialkyl azodicarboxylate, to give the mixed diester 3.2. The R ¹ substituent is varied by cleavage, using the methods described previously, to afford the monoacid product 3.3. The product is then 15 coupled, for example using methods described above, with the hydroxy compound R ³ OH, to give the diester product 3.4.

The procedure is illustrated in Scheme 3, Example 2. In this method, a monoallyl phosphonate 3.12 is coupled in tetrahydrofuran solution, in the presence of triphenylphosphine and diethylazodicarboxylate, with ethyl lactate 20 3.13 to give the mixed diester 3.14. The product is reacted with tris(triphenylphosphine) rhodium chloride (Wilkinson catalyst) in acetonitrile, as described previously, to remove the allyl group and produce the monoacid product 3.15. The latter compound is then coupled, in pyridine solution at ambient temperature, in the presence of dicyclohexyl carbodiimide, with one 25 molar equivalent of 3-hydroxypyridine 3.16 to yield the mixed diester 3.17.

Using the above procedures, but employing, in place of the ethyl lactate 3.13 or 3-hydroxyρyridine, a different hydroxyester 3.1 and/or a different hydroxy compound R ³ OH, the corresponding products 3.4 are obtained.

The mixed diesters 3.2 are also obtained from the monoesters 1.1 via the

30 intermediacy of the activated monoesters 3.5. In this procedure, the monoester

1.1 is converted into the activated compound 3.5 by reaction with, for example, phosphorus pentachloride, as described in J. Org. Chem., 2001, 66, 329, or with

155

5 thionyl chloride or oxalyl chloride (Lv = Cl), or with triisopropylbenzenesulfonyl chloride in pyridine, as described in Nucleosides and Nucleotides, 2000, 19, 1885, or with carbonyl diimidazole, as described in J. Med. Chem., 2002, 45, 1284. The resultant activated monoester is then reacted with the hydroxyester 3.1, as described above, to yield the mixed diester 3.2.

10 The procedure is illustrated in Scheme 3, Example 3. In this sequence, a monophenyl phosphonate 3.9 is reacted, in acetonitrile solution at 70°C, with ten equivalents of thionyl chloride, so as to produce the phosphoryl chloride 3.19. The product is then reacted with ethyl 4-carbamoyl-2-hydroxybutyrate 3.20 in dichloromethane containing triethylamine, to give the mixed diester 3.21.

15 Using the above procedures, but employing, in place of ethyl 4- carbamoyl-2-hydroxybutyrate 3.20, different hydroxyesters 3.1, the corresponding products 3.2 are obtained.

The mixed phosphonate diesters are also obtained by an alternative route for incorporation of the R ³ O group into intermediates 3.3 in which the

20 hydroxyester moiety is already incorporated. In this procedure, the monoacid intermediate 3.3 is converted into the activated derivative 3.6 in which Lv is a leaving group such as chloro, imidazole, and the like, as previously described. The activated intermediate is then reacted with the hydroxy compound R ³ OH, in the presence of a base, to yield the mixed diester product 3.4.

25 The method is illustrated in Scheme 3, Example 4. In this sequence, the phosphonate monoacid 3.22 is reacted with, trichloromethanesulfonyl chloride in tetrahydrofuran containing collidine, as described in J. Med. Chem., 1995, 38, 4648, to produce the trichloromethanesulfonyloxy product 3.23. This compound is reacted with 3-(morpholinomethyl)phenol 3.24 in dichloromethane containing

30 triethylamine, to yield the mixed diester product 3.25.

156

5 Using the above procedures, but employing, in place of with 3-

(morpholinonαethyl)phenol 3.24, different carbinols R ³ OH, the corresponding products 3.4 are obtained.

The phosphonate esters 3.4 are also obtained by means of alkylation reactions performed on the monoesters 1.1. The reaction between the monoacid

10 1.1 and the haloester 3.7 is performed in a polar solvent in the presence of a base such as diisopropylethylamine, as described in Anal. Chem., 1987, 59, 1056, or triethylamine, as described in J. Med. Chem., 1995, 38, 1372, or in a non-polar solvent such as benzene, in the presence of 18-crown-6, as described in Syn. Comm., 1995, 25, 3565.

15 The method is illustrated in Scheme 3, Example 5. In this procedure, the monoacid 3.26 is reacted with ethyl 2-bromo-3-phenylpropionate 3.27 and diisopropylethylamine in dimethylformamide at 80°C to afford the mixed diester product 3.28.

Using the above procedure, but employing, in place of ethyl 2-bromo-3-

20 phenylpropionate 3.27, different haloesters 3.7, the corresponding products 3.4 are obtained.

157

Scheme 3

R-link- -ffoR ¹

3.4 _(R4 °

A ^fcθ2R5

I Ha-R ⁴ -COOR ⁵

I P ³ - . ⁷ HO-R ⁴ -COOR ⁵ .0 R-IiOk-R _x -OR ¹ ^ R-link— 1(-OR ¹ . p p

R-link — P.-OH *~ R-link — F(-OR ³

OH 3.1 O-R ⁴ -COOR ⁵ O-R ⁴ -COOR ⁵ O-R ⁴ -COOR ⁵

1.1 ^ 3.2 3.3 3.4

3.1

O

R-link — f(-OR ¹ R-link — R^-Lv

O-R ⁴ -COOR ⁵ 3.5 ^LV 3.6

Scheme 3 Example 1

R-link — F^-OPh

R.πnk-ff P OPh HOCH ₂ CH(Me)CO ₂ Et OH 3.10

CO ₂ Et

3.9 Me ^

3.11

Scheme 3 Example 2

p HOCH(Me)CO ₂ Et O O

R-liπk — R-O »- R-link — P.-O - Cr ^OH

OH^- \ 3.13 o ^V -

\

Me— { Me— ( Me— ( N

CO ₂ Et CO ₂ Et CO ₂ Et

³ - ¹² 3.14 3.15 3.17

Scheme 3 Example 3 p EtO ₂ CCH(OH)CH ₂ CH ₂ CONH _{2 Q}

R-link — P _x -OPh SOCI _{2 R} _l _ink _p'°OPh 3.20 _λ R-link— ff OPh OH 3.18

Cl

CO ₂ Et

3.9 3.19 H ₂ N 3.21

158

Scheme 3 Example 4 p p ^r v ^ϋ o

R-link — R-OH *~ R-link — R-OSO ₂ CCI ₃ »~ R-link — R-O p p 3.24 p

Me— ( Me— {

CO ₂ Et CO ₂ Et

3.22 3.23 3.25

Scheme 3 Example 5

P BrCH(Bn)CO ₂ Et P

_R _| _ink __f£-OH 1— U^ R-|ink— F(-OCH(Bn)CO ₂ Et

OCH ₂ CF ₃ 3.27 OCH ₂ CF ₃ 3.26 3.28

Scheme 4 illustrates methods for the preparation of phosphonate diesters in which both the ester sύbstituents incorporate carboalkoxy groups.

10 The compounds are prepared directly or indirectly from the phosphonic acids 1.6. In one alternative, the phosphonic acid is coupled with the hydroxyester 4.2, using the conditions described previously in Schemes 1 - 3, such as coupling reactions using dicyclohexyl carbodiimide or similar reagents, or under the conditions of the Mitsonobu reaction, to afford the diester product

15 4.3 in which the ester substituents are identical.

This method is illustrated in Scheme 4, Example 1. In this procedure, the phosphonic acid 1.6 is reacted with three molar equivalents of butyl lactate 4.5 in the presence of Aldrithiol-2 and triphenyl phosphine in pyridine at ca. 70 ⁰ C, to afford the diester 4.6.

20 Using the above procedure, but employing, in place of butyl lactate 4.5, different hydroxyesters 4.2, the corresponding products 4.3 are obtained.

159

5 Alternatively, the diesters 4.3 are obtained by atkylation of the phosphonic acid 1.6 with a haloester 4.1. The alkylation reaction is performed as described in Scheme 3 for the preparation of the esters 3.4.

This method is illustrated in Scheme 4, Example 2. In this procedure, the phosphonic acid 1.6 is reacted with excess ethyl 3-bromo-2-methylpropionate 4.7 10 and diisopropylethylamine in dimethylformamide at ca.80°C, as described in Anal. Chem., 1987, 59, 1056, to produce the diester 4.8.

Using the above procedure, but employing, in place of ethyl 3-bromo-2~ methylpropionate 4.7, different haloesters 4.1, the corresponding products 4.3 are obtained.

15 The diesters 4.3 are also obtained by displacement reactions of activated derivatives 1.7 of the phosphonic acid with the hydroxyesters 4.2. The displacement reaction is performed in a polar solvent in the presence of a suitable base, as described in Scheme 3. The displacement reaction is performed in the presence of an excess of the hydroxyester, to afford the diester product 4.3 20 in which the ester substituents are identical, or sequentially with limited amounts of different hydroxyesters, to prepare diesters 4.3 in which the ester substituents are different.

The methods are illustrated in Scheme 4, Examples 3 and 4. As shown in Example 3, the phosphoryl dichloride 2.22 is reacted with three molar 25 equivalents of ethyl 3-hydroxy-2-(hydroxymethyI)propionate 4.9 in tetrahydrofuran containing potassium carbonate, to obtain the diester product 4.10.

Using the above procedure, but employing, in place of ethyl 3-hydroxy-2- (hydroxymethyl)propionate 4.9, different hydroxyesters 4.2, the corresponding 30 products 4.3 are obtained.

Scheme 4, Example 4 depicts the displacement reaction between equimolar amounts of the phosphoryl dichloride 2.22 and ethyl 2-methyl-3-

160

5 hydroxypropionate 4.11, to yield the monoester product 4.12. The reaction is conducted in acetonitrile at 70°C in the presence of diisopropylethylamine. The product 4.12 is then reacted, under the same conditions, with one molar equivalent of ethyl lactate 4.13, to give the diester product 4.14. Using the above procedures, but employing, in place of ethyl 2-methyl-3- 10 hydroxypropionate 4.11 and ethyl lactate 4.13, sequential reactions with different hydroxyesters 4.2, the corresponding products 4.3 are obtained.

161

Scheme 4

P

R-link — f^-OH — R-link — f(-Lv

O(R ⁴ )CO ₂ R ⁵ O(R ⁴ )CO ₂ R ⁵ 4.5 4.4

4.2 4.1 4.2

HO(R ⁴ JCO ₂ R ⁵

R-link— F(-OH 4.2 R-link— R^-O(R ⁴ JCO ₂ R ⁵ 1.6 OH 0(R ⁴ JCO ₂ R ⁵

Ha(R ⁴ JCO ₂ R ⁵ 4.3

4.2

O

R-link — P^-Lv R-link — P^-Lv Lv 4.2 0(R ⁴ JCO ₂ R ⁵

1.7 4.4 Scheme 4 Example 1

,P HOCH(CH ₃ )CO ₂ Bu .O

R-link — P _x -OH ... _R .| _ink — #- OCH(CH ₃ )CO ₂ Bu

OH 4.5 OCH(CH ₃ )CO ₂ Bu

1.6 4.6

Scheme 4 Example 2 p BrCH ₂ CH(CH ₃ )CO ₂ Et O l-link — P^-OH ^. R-iink — R^-OCH ₂ CH(CH ₃ )CO ₂ Et

OH 4.7 OCH ₂ CH(CH ₃ )CO ₂ Et ^" 1.6 4.8 Scheme 4 Example 3

(HOCH ₂ ) ₂ CHCO ₂ Et

.O

R-link — F(-CI R-link— R^-OCH ₂ CH(CH ₂ OH)CO ₂ Et Cl 4.9 OCH ₂ CH(CH ₂ OH)CO ₂ Et

2.22 4.10 Scheme 4 Example 4 _o HOCH ₂ CH(CH ₃ )CO ₂ Et _o HOCH(CH ₃ )CO ₂ Et _o

R-link — P^-Cl * ^► R-link — R-OCH ₂ CH(CH ₃ )CO ₂ Et »» R-link— R^-OCH ₂ CH(CH ₃ )CO ₂ Et

4 1

Cl ⁴ -. ¹ 1 ¹ h Cil ⁴ - ¹³ OCH(CH ₃ )CO ₂ Et 2.22 4.12 4.14

162

The compounds of Formula I-IV include all stereoisomers, and mixtures thereof. For example and not by way of limitation, the compounds of Formula I include at least the following stereoisomers:

,1 wy B ¹

P L ² -

_R 3aV'- \ / '""// _R 8 W R ₃₈ ιWVv>'T T""///R8

^R4 *T .P" ^R7 pj4\W"'i 1"'"///R7

OR ^a OR ^a OR ^a OR ^a

D1

^ JK jF "^ f-ι _llR % W ²

10

OR ^a OR ^a OR ^a OR ^a

■°v . -e,

W ² ^ _o R ₃ ³ _a ^' '"/ _/ R8 W z' _R 3a """//R8

Ft" I ^R ⁷ R4tt»'"' ϊ"""//R7

OR ^a OR ^a OR ^a OR ^a

O O

B ¹ -4 .B ¹

W ^2' _R 3a*^\ /"""// _R 8 W ² ' _R 33aa«»^f . y^Rβ

_R 4W-J |^ _R 7 R ⁴ »4 1"""//R7

OR ^a OR ^a OR ^a OR ^a

163

O. B ¹

8 W ² _R 3a^λ r S** _R 8

OR ^a OR ^a OR ^a OR ^a

W ²

In another embodiment, the compounds of Formula I-IV are selected from the group consisting of

HO? ^■ =bi =N

JO. /= /= N N. NH,

^

N^ N

HO OH HO OH

KJ

O. O' K

N; .N

10 X ^P HOτ "OH ^{r^}

O O Il

O

HO ^' >> ^ _M A?v ^V O.V n =N

OH HO

/ N. NH,

HO ^

^

N

HO OH

^{K '} r HO ^± OH ^C

164

r ==N

HN-«ι A O _s ^N NH ₂ HN'

>^ /

N^ N

HO t>H

O

Ay*

^NH ₂ ^HO ^- _^ / ^O V/ ^O N^N-i _o n 1 π ^{: =?v} ^ — - w _^ ^ ^N -

V ^ Hd ^H ^N ^ _f OH

O

P

\ O

HCX" Q Ov

* * O

HO "OH

HO OH

P

O O W Il o

HO' ^O ^O"°^_,°V I NH / ^O ONH

HO O

^

HO OH

P

O

HO 'OH

^ ^NH

HN. " ^/O^ .O,

10

NH ₂ / ^NH N

HN U Q HN :x!r .NN--λ4 O E^ .CL -N~4

^ O

— f

HO "OH HO OH

NH ₂

(

HoJ? Q N

O. HO- ^P ^

HO OH

NH ₂ NH ₂

/ \

HO.2 O.i

A HO' ^K ~ lX!r N-4 *

O

HO OH

HO OH

NH ₂

O O

W » ^

N

HO^O^O ?^O O. LN— 4 HO ^ O

HO OH

NH ₂ f -N

H ^N N^-°- ^λ N— 4

^ O

HO ^" OH

NH ₂

J O / N

HN M ^ Q HN ⁰ V _Z ONH

HO OH

10

166

NH ₂

I o

CL ( ⁴ N

HN' ^P ^ ^»N -i HO.° 'I n N IN==NN

V> =^w O HO

HO OH N; .N

OH

o" ^p "^ ^' \!r ^N Vυ ^NH2

OH ^^

O O W Il

O

^HO '^ _H ^O _H άV N=N

/

HO ^' \> ^N' γ N^S^Nr ^NH2

OH

HN.§ o

OH

H ₂ N y=N H ₂ N

>=N

^{^} V^ ^O yN.^ N^\ ^

OH

OH

H ₂ N

>=N

HN.? o N^ )

10

167

H ₂ N,

O \ y=s=-sNiN.

« Il .Px o N* /) Hθj? .Q,

KJ* // N NH ₂

^■ N HO' rv<° t)H

-OH

\ O

V r 'OH

::^ °- t)H

O W 9

HO ^' ^^ ^V NCV^NH,

HO

I)H

O ^ /=N

HO.u^o. OH

HO ^' X < λ rV ^y i

NH ₂

HO.JI

=N HO'

HO _; ^O O _N -N. NH ₂

^ N; N NH ₂

OH

10

168

OH

HoJ? =N

.

NH ₅ HO WO. v

N; .N

Il OH

\-oi =N

.O.

NH ₂ P-^S NH ₂ N; N OH

Il OH T T Il ^ OH T

NH ₂ T NH ₂

HN. I o /= ^N OH

NHp

OH

10 NHP

169

HN' ^P ^

NH,

NH _?

and

or pharmaceutically acceptable salts, solvates, and/or esters thereof. ^• 10 In another embodiment of the compounds of Formula I- IV, -U-L ² -

P(O)WiW ² is -0-C(R ^1O ) ₂ -P(O)W ¹ W ² , wherein each R ¹⁰ is independently H, alkoxy, alkyl, or halo. Non-limiting examples of suitable -0-C(R ¹⁰ ^-P(O)W ¹ W ² groups include -0-CH ₂ -P(O) W ¹ W ² , -CHCl-O-P(O)W ¹ W ² , -CCb-O-P(O)W ¹ W ² , - CH(CHS)-O-P(O)W ¹ WVC(CHS) ₂ -O-P(O)W ¹ W ² , etc.

170

5 In another embodiment of the compounds of Formula I-IV, -U-L ² -

P(O)W ¹ W ² is -S-C(R ^l0 )2-P(O)W ¹ W ² , wherein each R ¹⁰ is independently H ₇ alkoxy, alkyl, or halo. Non-limiting examples of suitable -S-C(R ¹⁰ ^-P(O)W ¹ W ² groups include -S-CH ₂ -P(O)W ¹ W ² , -S-CHCl-P(O)W ¹ W ² , -S-CHF-P(O)W ¹ W ² , -S- CH(CHs)-P(O)W ¹ W ² ^S-CF ₂ -P(O)W ¹ W ² , etc.

10 In another embodiment of the compounds of Formula I-IV, -U-L ² -

P(O)W ¹ W ² is -N(R ¹¹ J-C(R ^1O ) ₂ -P(O)W ¹ W ² , wherein each R ¹⁰ is independently H, alkoxy, alkyl, or halo and R" is H, alkyl, aryl, or substituted aryl. Non-limiting examples of suitable -N(R ¹¹ J-C(R ^1O ) ₂ -P(O)W ¹ W ² groups include -NH-CH ₂ - P(O)W ¹ W ² , -NH-CHCl-P(O)W ¹ W ² , -NH-CHF-P(O)W ¹ W ² , -NH-CH(CHs)-

15 P(O)W ¹ W ² , -NH-CF ₂ -P(O)W ¹ W ² , — N(CHs)-CH ₂ -P(O)W ¹ W ² , -N(CHs)-CHCl- P(O)W ¹ W ² , -N(CHs)-CF ₂ -P(O)W ¹ W ² , - N(CHs)-CH(CHs)-P(O)W ¹ W ² , - N(CH ₃ )- CHF-P(O)W ¹ W ² , - N(phenyl)-CH ₂ -P(O) W ¹ W ² , - N(phenyl)-CHCl-P(O)W ¹ W ² , - N(phenyl)-CF ₂ -P(O) W ¹ W ² , - N(PhBn ₇ I)-CH(CHs)-P(O)W ¹ W ² , etc.

One skilled in the art will recognize that nucleobases can exist in

20 tautomeric forms. For example, structures (a) and (b) can have equivalent tautomeric forms as shown below:

(a) and

OH

_F χ

(b)

171

5 AU possible tautomeric forms of the nucleobases of all of the embodiments are within the scope of the invention.

By way of example and not by limitation, nucleobases B ¹ , B ² , or B ³ for Formula I-IV that are within the scope of the invention include:

VV Ύ ^NH2 V ^V V ¹

r ^N y

V M _^ τ N ^NH2 v

T _Nl γN

N, V

10 ^N ^'\ N NHH _{2 f} /^ ₇ π Cl

W r ^N γ^r ^{^π} ^N^

OCH ₃

NH ₂ / ^N -- όl N^- ¹ N

O _C H ₃ ^NCV^OCHa _^ ^ ^ _^QCH

N^N ^N γ ^N J ₁ ^N

N^N O I _{1 N} ^

Cl Nc^N O Jj ₁ ^

172

V ^{Q ^ t} - ^{M r ^ tr^} .«. v

NH ₂ ' J"

>, .N r -Y N. ^f i Ϋ NH ₂

^^/ ^N

Cl

H

V ^{ς^} r ^% "^ _r ^% S *-o

Cl

%' ^{ς^} o v ^% c» . ^a o -

NH ₂ ' N^

N

l-v^ rY \ vV λ v--Dγ°"

/

NH ₂

173

N=π

V ^~ V I ^N M=~

I ι Irl ^OH v r ^~ O \<rv Cτ ^OH =\

1 U

N.

NH ₂ *-V NC _{N ^} r OH N

/ ^N -s

N Cl

==zλ NCN^N

V OCH,

N^^N V OCH, N~ --N,

T N^γN \ OCH,

NH _{2 ^} N _^

C/

NHo

N Ac-s _^ ¹ NA O fl N^ TN 0 T F

NH _{2 ^ N^}

N=r

^N ^ _y N O ; ^N =\ I ' ''^ - ^l Cl 1 U Il N- _^ .w 0

N; .N O N<γN O

I .

/ NV;

=λ NH ₂

N — i , N^s _^ N O

' A H

N^M O

10 NH^

Nz

*" ^φ YV N=*

N^N O N=r Cf V

N^ _^ N V V

NH ₂

174

N=

=\ H

N=r\

/ ci iLJiij ¹ ^

N=r

=\ H N=

=\ H π ^p ro ^W o v ^{J^} o

Cl

/=N

.NH, s-N A _{^ -} NH ₂ ^ r T ^ r T

NCN^N w NwH ₂ _ .N. y

N

\

^■" *- -, NH ₂ ^ ^N^

/=N

10 Cl _^ N^ _/ N T T

NH ₂ ^N^

3

^N Y Cl ^N ^ ^N γ N^ ^h _^ Nr ^{OCH3 X} T

NJH ₂ ^X T ^NX^ r=N

^c ' N^N O T

^^ , NH ₂

175

=N

YΎ ,°oΎ> .NH, =N

W N^YN "

/' V 11

O

N ^? cs _Ύ ^V λ _^ *r°Y O ^N /= ^N , N

T / N ^M O

N. T Cl

=N H

-:N O^ ^N

^V Y ^Y TT

NH ₂

/= ^N v H

-N. X _^ O^ ^N. /=N H v ^N γ ^h r°γ ^N

Cl _^ r ^N γ N-^hNr ^N ^7

.r^

V

NH ₂

Cl

/= ^N _V H ' N<^N ¹ ^- ^/

10 N

INπ ₂ ^ :N

. H N=N N=N

^cι , ^N ^ ^N , IH _{2 ^ /} .)N'.

176

, ^N = ^N . N=N N=N

W " w W Y^ ^W ^

Cl N. -N T MLJ T T

/ N NI.

N=N N=N N=N

W -* N=N OH

N^N V

Cl N^^N

NH ₂

N=N

N=N N=N

W Y" N=N

OCH, \

%- v ^~N V OCH-,

N^vN ^OCH3 ^ ^N VY

Cl N. ,N

NH ₉

N=N

N=N

OCH^

N=N O^ ^NH ₂

%'

N^N

Cl T NH ₂

N=N

^N _. λ^ _/ O^^NH ₂ N=N

O^ _/ NH ₂

N N==NN \ N=N \

^V -V N^ _/ Y ^V N 0 ^* W _N Y ^{1 V} M 0 V- ^O $ r

_N W u » 0

10 T γi

NH ₂ T N.

/ Cl

N=N H

N=N

N^ _/ N O K^ T

^^ , NH ₂

177

' ^N=N v H N=N

V-V vY Hi - ^feN , H

/ ^N ^ Cl

N=N _H N=N

/ N=N _V H

T ^{^} V

NH ₂ T

Cl

N=N N=N

%- ^ςv Nc^Nt ^-> /

, * ^< " T NH ^{^} ₃ ^ ^W T N. O-

-K

, ^N=N v H \

< ^

N-N N-N

^■N W ^N χ> N-N NH ₅

N NH,

T W ^N' VN" NH ₅

N^N

Cl N; N

NH ₂

A N-N -<

N.jL _^ NH ₂ λ N-N / N-N χl A

N-N NH, N-N

N ^' V

N<^N N, <N

.N . \\

Ncy WN N^-N . \\ Cl N; \ NH,

Cl

^

N-N A N-N A

N-N N ^ OH N. ^-v ϊ OH N-N

^N' <Jγ ^OH . \\ N<3N^OH N-^N N .^ WN y UN

10 N^^N N . _c

NH ₂ a

178

N-N

<

N-N OCH, OCH, -K N-N

N^Jk^OCH ₃ < _s λγOCH ₃

1 N^N

N N Nc ₅ ^N N. y Cl

λ

A N-N A N-N

N-N >N

"SV NH,

N^N O ^{■2 N'} vV°γ ^NH2

N^^Nγ NH,

-K N-N

,NH ₂ ^ N-N \

N-N \

N^N O ^N V N^VN ^O Y O ^N - C! W N. .N O NH ₂

•A N-N I ^ N.-N

NcyN O ¹ V N^VY N O ^■ ^; N-.

Cl ^N V N^VN O

^:

N-N H N-N H

N^ _/ N O ^NW .O _^ y ^Nt.

NH ₂

^; N-N

^N' W N^N°Y X O

10 Cl ^N W ^N ^7 ^V

179

A N-N -^:

H N M--NN LJ

^• ^

N-N

N^N

N^N H.

N ^ _/ N ^W

/

λ-N H V« H . λ.-N

W O - W ^~ O W o -

NH ₂ / / ^N -- / Cl / _e C _t l _c L.

In still another embodiment, the compounds of Formula I, Formula II, Formula III, or Formula IV are named below in tabular format (Table 6) as compounds of general Formula V: wl °

w ^2' I CHγ —ex

XT" ^" L /

/ _^ -OH

)

10 X2 \ ₃

Formula V wherein Xl, X2, X3, and L represent substituents attached to the tetrahydrofuranyl ring as defined in Tables 1-4, below; B is a purine defined in Table 5, below; and each W ¹ and W ² are as previously defined above.

15 The point of attachment of the core structure C is indicated in each of the structures of Xl, X2, X3, L and B. Each structure in Tables 1-5 is represented by an alphanumeric ""code". Each structure of a compound of Formula V can thus be designated in tabular form by combining the "code" representing each structural moiety using the following syntax: X1.X2.X3.L.B. Thus, for example,

20 Xla.X2c.X3a.Ll.Bl represents the following structure:

180

amino halo

Table 1: Xl Structures

Table 2: X2 Structures

10

Table 3: X3 Structures

Table 4: L Structures

15

181

Table 5: B Structures

Table 6: List of Compounds of Formula V

Xla.X2a.X3a.Ll.Bl, Xla.X2a.X3a.Ll.B2, Xla.X2a.X3a.Ll.B3, Xla.X2a.X3a.Ll.B4,

Xla.X2a.X3a.L2.Bl, Xla.X2a.X3a.L2.B2, Xla.X2a.X3a.L2.B3, Xla.X2aJC3a.L2.B4,

10 Xla.X2a.X3a.L3.Bl, Xla.X2a.X3a.L3.B2, Xla.X2a.X3a.L3.B3, Xla.X2a.X3a.L3.B4, Xla.X2a.X3b.Ll.Bl, Xla.X2a.X3b.Ll.B2, XlaJC2a.X3b.Ll.B3, Xla.X2a.X3b.Ll.B4, Xla.X2a.X3b.L2.Bl, Xla.X2a.X3b.L2.B2, Xla.X2a.X3b.L2.B3, Xla.X2a.X3b.L2.B4, Xla.X2a.X3b.L3.Bl, Xla.X2a.X3b.L3.B2, Xla.X2a.X3b.L3.B3, Xla.X2a.X3b.L3.B4, Xla.X2a.X3c.Ll.Bl, Xla.X2a.X3c.Ll.B2, Xla.X2a.X3c.Ll.B3, Xla.X2a.X3c.Ll.B4,

15 Xla.X2a.X3c.L2.Bl, Xla.X2a.X3c.L2.B2, Xla.X2a.X3c.L2.B3, Xla.X2a.X3c.L2.B4, Xla.X2a.X3c.L3.Bl, Xla.X2a.X3c.L3.B2, Xla.X2a.X3c.L3.B3, Xla.X2a.X3c.L3.B4, Xla.X2b.X3a.Ll.Bl, Xla.X2b.X3a.Ll.B2, XlaJC2b.X3a.Ll.B3, Xla.X2b.X3a.Ll.B4, Xla.X2b.X3a.L2.Bl, Xla.X2b.X3a.L2.B2, Xla.X2b.X3a.L2.B3, Xla.X2b.X3a.L2.B4,

182

5 Xla.X2b.X3a.L3.Bl, Xla.X2b.X3a.L3.B2, Xla.X2b.X3a.L3.B3, Xla.X2b.X3a.L3.B4, Xla.X2b.X3b.Ll.Bl, Xla.X2b.X3b.Ll.B2, Xla.X2b.X3b.Ll.B3, Xla.X2b.X3b.Ll.B4, Xla.X2b.X3b.L2.Bl, Xla.X2b.X3b.L2.B2, Xla.X2b.X3b.L2.B3, Xla.X2b.X3b.L2.B4, Xla.X2b.X3b.L3.Bl, Xla.X2b.X3b.L3.B2, Xla.X2b.X3b.L3.B3, Xla.X2b.X3b.L3.B4, Xla.X2b.X3c.Ll.Bl, Xla.X2b.X3c.Ll.B2, Xla.X2b.X3c.Ll.B3, Xla.X2b.X3c.Ll.B4,

10 Xla.X2b.X3c.L2.Bl, Xla.X2b.X3c.L2.B2, Xla.X2b.X3c.L2.B3, Xla.X2b.X3c.L2.B4, Xla.X2b.X3c.L3.Bl, Xla.X2b.X3c.L3.B2, Xla.X2b.X3c.L3.B3, Xla.X2b.X3c.L3.B4, Xla.X2c.X3a.Ll.Bl, Xla.X2c.X3a.Ll.B2, Xla.X2c.X3a.Ll.B3, Xla.X2c.X3a.Ll.B4, Xla.X2c.X3a.L2.Bl, Xla.X2c.X3a.L2.B2, Xla.X2c.X3a.L2.B3, Xla.X2c.X3a.L2.B4, Xla.X2c.X3a.L3.Bl, Xla.X2c.X3a.L3.B2, Xla.X2c.X3a.L3.B3, Xla.X2c.X3a.L3.B4,

15 Xla.X2c.X3b.Ll.Bl, Xla.X2c.X3b.Ll.B2, Xla.X2c.X3b.Ll.B3, Xla.X2c.X3b.Ll.B4, Xla.X2c.X3b.L2.Bl, Xla.X2c.X3b.L2.B2, Xla.X2c.X3b.L2.B3, Xla.X2c.X3b.L2.B4, Xla.X2c.X3b.L3.Bl, Xla.X2c.X3b.L3.B2, Xla.X2c.X3b.L3.B3, Xla.X2c.X3b.L3.B4, Xla.X2c.X3c.Ll.Bl, Xla.X2c.X3c.Ll.B2, Xla.X2c.X3c.Ll.B3, Xla.X2c.X3c.Ll.B4, Xla.X2c.X3c.L2.Bl, Xla.X2c.X3c.L2.B2, Xla.X2c.X3c.L2.B3, Xla.X2c.X3c.L2.B4,

20 Xla.X2c.X3c.L3.Bl, Xla.X2c.X3c.L3.B2, Xla.X2c.X3c.L3.B3, Xla.X2c.X3c.L3.B4, Xlb.X2a.X3a.Ll.Bl, Xlb.X2a.X3a.Ll.B2, Xlb.X2a.X3a.Ll.B3, Xlb.X2a.X3a.Ll.B4, Xlb.X2a.X3a.L2.Bl, Xlb.X2a.X3a.L2.B2, Xlb.X2a.X3a.L2.B3, Xlb.X2a.X3a.L2.B4, Xlb.X2a.X3a.L3.Bl, Xlb.X2a.X3a.L3.B2, Xlb.X2a.X3a.L3.B3, Xlb.X2a.X3a.L3.B4, Xlb.X2a.X3b.Ll.Bl, Xlb.X2a.X3b.Ll.B2, Xlb.X2a.X3b.Ll.B3, Xlb.X2a.X3b.Ll.B4,

25 Xlb.X2a.X3b.L2.Bl, Xlb.X2a.X3b.L2.B2, Xlb.X2a.X3b.L2.B3, Xlb.X2a.X3b.L2.B4, Xlb.X2a.X3b.L3.Bl, Xlb.X2a.X3b.L3.B2, Xlb.X2a.X3b.L3.B3, Xlb.X2a.X3b.L3.B4, Xlb.X2a.X3c.Ll.Bl, Xlb.X2a.X3c.Ll.B2, Xlb.X2a.X3c.Ll.B3, Xlb.X2a.X3c.Ll.B4, Xlb.X2a.X3c.L2.Bl, Xlb.X2a.X3c.L2.B2, Xlb.X2a.X3c.L2.B3, Xlb.X2a.X3c.L2.B4, Xlb.X2a.X3c.L3.Bl, Xlb.X2a.X3c.L3.B2, Xlb.X2a.X3c.L3.B3, Xlb.X2a.X3c.L3.B4,

30 Xlb.X2b.X3a.Ll.Bl, Xlb.X2b.X3a.Ll.B2, Xlb.X2b.X3a.Ll.B3, Xlb.X2b.X3a.Ll.B4, Xlb.X2b.X3a.L2.Bl, Xlb.X2b.X3a.L2.B2, Xlb.X2b.X3a.L2.B3, Xlb.X2b.X3a.L2.B4, Xlb.X2b.X3a.L3.Bl, Xlb.X2b.X3a.L3.B2, Xlb.X2b.X3a.L3.B3, Xlb.X2b.X3a.L3.B4,

183

5 Xlb.X2b.X3b.Ll.Bl ₇ Xlb.X2b.X3b.Ll.B2, Xlb.X2b.X3b.Ll.B3, Xlb.X2b.X3b.Ll.B4, Xlb.X2b.X3b.L2.Bl, Xlb.X2b.X3b.L2.B2, Xlb.X2b.X3b.L2.B3, Xlb.X2b.X3b.L2.B4, Xlb.X2bJC3b.L3.Bl, Xlb.X2b.X3b.L3.B2, Xlb.X2b.X3b.L3.B3, Xlb.X2b.X3b.L3.B4, Xlb.X2b.X3c.Ll.Bl, Xlb.X2b.X3c.Ll.B2, XlbJC2b.X3c.Ll.B3, Xlb.X2b.X3c.Ll.B4, Xlb.X2b JC3c.L2.Bl, Xlb.X2b.X3c.L2.B2, Xlb.X2b.X3c.L2.B3, Xlb.X2b.X3c.L2.B4,

10 Xlb.X2b.X3c.L3.Bl, Xlb.X2b.X3c.L3.B2, Xlb.X2b.X3c.L3.B3, Xlb.X2b.X3c.L3.B4, Xlb.X2c.X3a.Ll.Bl, Xlb.X2c.X3a.Ll.B2, Xlb.X2c.X3a.Ll.B3, Xlb.X2c.X3a.Ll.B4, XIb JC2cJC3a.L2.Bl, Xlb.X2c.X3a.L2.B2, Xlb.X2c.X3a.L2.B3, Xlb.X2c.X3a.L2.B4, Xlb.X2c.X3a.L3.Bl, Xlb.X2c.X3a.L3.B2, Xlb.X2c.X3a.L3.B3, Xlb.X2c.X3a.L3.B4, Xlb.X2cJC3b.Ll.Bl, Xlb.X2c.X3b.Ll.B2, Xlb.X2c.X3b.Ll.B3, Xlb.X2c.X3b.Ll.B4,

15 Xlb.X2cJC3b.L2.Bl, Xlb.X2c.X3b.L2.B2, Xlb.X2c.X3b.L2.B3, Xlb.X2c.X3b.L2.B4, XIb JC2cJC3b.L3.Bl, Xlb.X2c.X3b.L3.B2, XIb JC2cJC3b.L3.B3, Xlb.X2c.X3b.L3.B4, Xlb.X2c.X3c.Ll.Bl, Xlb.X2c.X3c.Ll.B2, Xlb.X2c.X3c.Ll.B3, XlbJC2cJC3c.Ll.B4, Xlb.X2c.X3c.L2.Bl, Xlb.X2c.X3c.L2.B2, Xlb.X2c.X3c.L2.B3, Xlb.X2c.X3c.L2.B4, Xlb.X2cJC3c.L3.Bl, Xlb.X2c.X3c.L3.B2, Xlb.X2c.X3c.L3.B3, Xlb.X2c.X3c.L3.B4,

20 Xlc.X2a.X3a.Ll.Bl, Xlc.X2a.X3a.Ll.B2, Xlc.X2a.X3a.Ll.B3, Xlc.X2a.X3a.Ll .B4, Xlc.X2a.X3a.L2.Bl, Xlc.X2a.X3a.L2.B2, Xlc.X2a.X3a.L2.B3, Xlc.X2a.X3a.L2.B4, Xlc.X2a.X3a.L3.Bl, Xlc.X2a.X3a.L3.B2, Xlc.X2a.X3a.L3.B3, Xlc.X2a.X3a.L3.B4, Xlc.X2a.X3b.Ll.Bl, Xlc.X2a.X3b.Ll.B2, Xlc.X2a.X3b.Ll.B3, Xlc.X2a.X3b.Ll.B4, Xlc.X2aJC3b.L2.Bl, Xlc.X2a.X3b.L2.B2, Xlc.X2a.X3b.L2.B3, Xlc.X2a.X3b.L2.B4,

25 Xlc.X2a.X3b.L3.Bl, Xlc.X2a.X3b.L3.B2, Xlc.X2a.X3b.L3.B3, Xlc.X2a.X3b.L3.B4, XlcJC2aJC3c.Ll.Bl, Xlc.X2a.X3c.Ll.B2, Xlc.X2a.X3c.Ll.B3, Xlc.X2a.X3c.Ll.B4, Xlc.X2a.X3c.L2.Bl, Xlc.X2a.X3c.L2.B2, Xlc.X2a.X3c.L2.B3, Xlc.X2a.X3c.L2.B4, Xlc.X2a.X3c.L3.Bl, Xlc.X2a.X3c.L3.B2, Xlc.X2a.X3c.L3.B3, Xlc.X2a.X3c.L3.B4, Xlc.X2bJC3a.Ll.Bl, Xlc.X2bJOa.Ll.B2, Xlc.X2b.X3a.Ll.B3, Xlc.X2b.X3a.Ll.B4,

30 Xlc.X2b.X3a.L2.Bl, Xlc.X2b.X3a.L2.B2, Xlc.X2b.X3a.L2.B3, Xlc.X2b.X3a.L2.B4, Xlc.X2b.X3a.L3.Bl, Xlc.X2b.X3a.L3.B2, Xlc.X2b.X3a.L3.B3, Xlc.X2b.X3a.L3.B4, Xlc.X2b.X3b.Ll.Bl, Xlc.X2b.X3b.Ll.B2, Xlc.X2b.X3b.Ll.B3, Xlc.X2b.X3b.Ll.B4,

184

5 Xlc.X2b.X3b.L2.Bl, Xlc.X2b.X3b.L2.B2, Xlc.X2b.X3b.L2.B3, Xlc.X2b.X3b.L2.B4, Xlc.X2b.X3b.L3.Bl, Xlc.X2b.X3b.L3.B2, Xlc.X2b.X3b.L3.B3, Xlc.X2b.X3b.L3.B4, Xlc.X2b.X3c.Ll.Bl, Xlc.X2b.X3c.Ll.B2, Xlc.X2b.X3c.Ll.B3, Xlc.X2b.X3c.Ll.B4, Xlc.X2b.X3c.L2.Bl, Xlc.X2b.X3c.L2.B2, Xlc.X2b.X3c.L2.B3, Xlc.X2b.X3c.L2.B4, Xlc.X2b.X3c.L3.Bl, Xlc.X2b.X3c.L3.B2, Xlc.X2b.X3c.L3.B3, Xlc.X2b.X3c.L3.B4,

10 Xlc.X2c.X3a.Ll.Bl, Xlc.X2c.X3a.Ll.B2, Xlc.X2c.X3a.Ll.B3, Xlc.X2c.X3a.Ll.B4, Xlc.X2c.X3a.L2.Bl, Xlc.X2c.X3a.L2.B2, Xlc.X2c.X3a.L2.B3, Xlc.X2c.X3a.L2.B4, Xlc.X2c.X3a.L3.Bl, Xlc.X2c.X3a.L3.B2, Xlc.X2c.X3a.L3.B3, Xlc.X2c.X3a.L3.B4, Xlc.X2c.X3b.Ll.Bl, Xlc.X2c.X3b.Ll.B2, Xlc.X2c.X3b.Ll.B3, Xlc.X2c.X3b.Ll.B4, Xlc.X2c.X3b.L2.Bl, Xlc.X2c.X3b.L2.B2, Xlc.X2c.X3b.L2.B3, Xlc.X2c.X3b.L2.B4,

15 Xlc.X2c.X3b.L3.Bl, Xlc.X2c.X3b.L3.B2, Xlc.X2c.X3b.L3.B3, Xlc.X2c.X3b.L3.B4, Xlc.X2c.X3c.Ll.Bl, Xlc.X2c.X3c.Ll.B2, Xlc.X2c.X3c.Ll.B3, Xlc.X2c.X3c.Ll.B4, Xlc.X2c.X3c.L2.Bl, Xlc.X2c.X3c.L2.B2, Xlc.X2c.X3c.L2.B3, Xlc.X2c.X3c.L2.B4, Xlc.X2c.X3c.L3.Bl, Xlc.X2c.X3c.L3.B2, Xlc.X2c.X3c.L3.B3, Xlc.X2c.X3c.L3.B4,

20 Phosphonate Embodiments of Compounds of Formula I-IV

By way of example and not limitation, the phosphonate embodiments of Formula I-IV may be represented by the general formula "MBF":

O

Sc-

^Pd ² Pd ¹

MBF

25 Each embodiment of MBF is depicted as a substituted nucleus (Sc). Sc is described in formulae A-G of Table 1.1 below, wherein Sc is a generic formula for a compound of Formula I, Formula II, Formula III, or Formula IV, and the point of attachement to -P(O)Pd ¹ Pd ² is indicated with a wavy line.

185

Table 1.1

OH

NH ₂ /^VV* N^NH ₂

OH

B

OH OH

N-

•* \^Q NH ₂

HO OH i? 5H

C D -

OH ' NH ₂

/T ]T ^{^} N _N '/ Tl ■>

H(5 OH HO OH

OH N>

N ₃ ³ λ_/

HO OH G

10 Pd ¹ and Pd ² are. each independently selected from species in Tables 20.1 to

20.37. The variables used in Tables 20.1-20.37 (e.g., W ³ , R ¹ , etc.) pertain only to Tables 20.1-20.37, unless otherwise indicated. Additional phosphonate groups are disclosed in U.S. patent publication No. 2004/100960, the entirety of which is incorporated herein by reference.

15 The variables used in Tables 20.1 to 20.37 have the following definitions:

R ¹ is independently H or alkyl of 1 to 18 carbon atoms;

186

5 R ² is independently H, R ¹ , R ³ or R ⁴ wherein each R ⁴ is independently substituted with 0 to 3 R ³ groups or taken together at a carbon atom, two R ² groups form a ring of 3 to 8 carbons and the ring may be substituted with 0 to 3 R ³ groups;

R ³ is R ^3a , R ^3b , R ³⁰ or R ^3d , provided that when R ³ is bound to a heteroatom, 10 then R ³ is R ^3c or R ^M ;

R ^3a is F, Cl, Br ₇ I, -CN, Ns or -NCh; R ^3b is Y ¹ ;

R ^3c is -R", -N(R*)(R ^X ), -SR*, -S(O)R", -S(O) ₂ R-, -S(O)(OR*), -S(O) ₂ (OR*), - OC(Y ^] )R*, -OC(Y ¹ JOR*, -OC(Y ¹ XN(RO(RO), -SC(V)R*, -SC(Y ¹ PR*, - 15 SC(Y ¹ XN(RO(R*)), -N(R*)C(Y ¹ )R ^X , -N(R*)C(V)OR ^X , or -N(R*)C(Y ¹ )(N(R")(R ^X )) ; -R ^3d is -C(V)R", -C(V)OR* or -C(Y ¹ )(N(R ^X )(R*));

R ⁴ is an alkyl of 1 to 18 carbon atoms, alkenyl of 2 to 18 carbon atoms, or alkynyl of 2 to 18 carbon atoms;

R ⁵ is R ⁴ wherein each R ⁴ is substituted with 0 to 3 R ³ groups;

20 R ^5a is independently alkylene of 1 to 18 carbon atoms, alkenylene of 2 to

18 carbon atoms, or alkynylene of 2-18 carbon atoms any one of which alkylene, alkenylene or alkynylene is substituted with 0-3 R ³ groups; W ³ is W ⁴ or W ⁵ ;

W ⁴ is R ⁵ , -C(Y ¹ JR ⁵ , -C(Y ¹ JW ⁵ , -SO ₂ R ⁵ , or -SO ₂ W ⁵ ;

25 W ⁵ is carbocycle or heterocycle wherein W ⁵ is independently substituted with 0 to 3 R ² groups;

187

O^

λ R ⁵

.o.

^" W ³

O

O

O.

λ ^■ R ¹

.0.

λ ^" R ⁴

O

O

XX.

λ Ok

λ 'CV^

H

O

O

.Ok

λ

O 8

O _^ .O^

λ

O

1S8

CB ₃

.CH ₃

CH ₃

CHβ

O

W

189

CH ₃

λ

W ^{3 <} K R ⁵

O

13 CH ₃

λ \ R ¹ o

15

λ °\ CKfe

O 17

CH ₃

λ

XH ₃

CH ₃ O

19

λ O. -CH ₃

O

18

190

191

5 Table 20.5

W ³

H

H ₃ C

λ CX ^CH ₃

O

.29

192

Table 20.6

H ₃ C

CH ₃

λ

XHj

O

33

193

Table 20.7

W ³ W ³

^ ^ xx λ

W ³ °\ R ⁵

O

34 W ³

λ °\ R ⁴ R ¹

O

36 R ⁵

λ °\ W ³

O

38

R ⁵ .

λ o.

^' R ⁴

O

40

194

Table 20.8

R ⁴ R ⁴

λ Q

R ⁵

O

43 R ⁴

λ °\ R ¹

O

45 R ¹

λ o

R ⁵

O

47

R ¹

λ \ R ¹

O 49

195

⁰ ^^R ⁵ ς

CX

^* W ³ H 51

H O

50

λ O^

-R ¹

λ .CX

-R ⁴

H O 53

H O

52

O _^

λ -CH ₃

O _^

λ Ή

H O 55

H O

54

λ ~<α

il O 57

£&*

λ

H O

56

196

x^s^a Q^λQ. CB ₃

O

,CB ₃

O O-

H 58 CH ₃

O

& 59

CH ₃

O CH ₃

λ

O

H 60

197

Table 2Oli CH ₃

CH ₃

λ

λ '°\ /°\ R ⁵

W ³ H O

62 CH ₃

CH ₃

λ

λ O. °\ R ^{1 ■ R 4 H O}

H O 64

63

CH ₃

CH ₃

λ

λ JL ,o. °\ CH ₃

η H O

H O 66

65

CH ₃

λ ^{^} CH ₃

CH ₃ H O

68

λ .CH ₃

H O

67

198

CH ₃ CH ₃

λ

XHj .

H O

71

199

5 Table2Ol3 B ₃ C

λ

H O

72

H ₃ C

λ O^

R ⁴

H O

74

H ₃ C.

CH ₃

λ ¹ .ex

Ή

H O

76

H ₃ C

.CH ₃

λ O.

H O

78

200

201

W ³

W ³

CX^

λ O λ R ⁵

W ³ H O

H 84 ⁿ 83 °

W ³

W ³

λ O.

^■ R ^α

λ XX

- _R 4 H O

86

H O

85 R ⁵ R ⁵ °\

λ °\ R ^a w ³ H O

H O

87

R ⁵

R ⁵

λ

λ °\ R ⁴ H O

90

H O

89

202

Table 20.16

R ⁴ R ⁴

λ °\ λ

W ³ °\ R ⁵

H

O H O

91 92

R ⁴ R ⁴

λ O. λ O\

R ⁴ R ¹

H

O H

93 O

94 R ¹ R ¹

λ °\ λ

W ^{3 0} S

R ⁵

H

O H O

95 96

R ¹ R ¹

^ °\ A

R ⁴ ^R ¹

H O

H O

97 98

203

λ ^R*

O^

Vr

R ³ O 100

R* O

99

λ °-^

λ

B ³ O 102

R ³ O

101

λ -°\ CH ₃

λ .α. ^* H

R° O 104

R ³ O

103

.O. -CH ₃

R ³ O 106

,CH ₃

λ

R ³ O

105

204

.CB ₃

O

CB ₃ o O

*e 107 λ

CB ₃

&

R* 108

CB ₃

CB ₃

*

O

B. ^d 109

205

Table 20.19

CH ₃ CH ₃

(^ ^ ,α A w ^{3 0} S R ⁵

R ³ O R ³ O

110 111

CH ₃ CH ₃

λ O λ

R ⁴ °\ R ¹

R ³ O R ^d

112 O

113

CH ₃ CH ₃

λ O << O

H CH ₃

R ³

O R ³ O

114 115

CH ₃

λ

^{^} CH ₃

CH ₃

R ³ O

117

^ O^^CHg

R ³ O

116

206

Table 20.20

R ³ O

118

CH*

CH ₃ «

λ o>

'CH ₃

R ³ O

120

207

5 Table 20.21 KbC

Q

W ³

R ³ O

λ o

R ⁴

R ³ O

H ₃ C. ¹²³

λ α

H

H ₅ C o 128

λ Ck^^CHg

R ³ O

127

208

R ³ O

129

H ₃ C

CH ₃

λ ^v Cϊ%

R ³ O

131

209

Table 20.23

W ³ W ³

λ °\ ti

W ³ °\ R ⁵

R ^d

O R ³ O

132 133

W ³ W ³

λ °\ ti

R ⁴ °\ R ¹

R ³ O

134 R ³ O

135

R ⁵ R ⁵

<i °\ λ α

W ³ R ⁵

R ³

O R ³ O

136 137

R ⁵ R ⁵ ti. °\ λ

R ⁴ °\ R ¹

R ³

O R ^ό . O

138 139

210

Table 20.24

R ⁴ R ⁴

λ Q λ O

W ³ R ⁵

R ³ O

140

R ⁴ R ⁴

λ °\ λ

R ⁴ °\ R ¹

R ³ O R ³ O

142 143

R ¹ R ¹

λ O. (<

W ³ °\ R ⁵

R ³ O R ³ O

144 145

R ¹ R ¹

<< O. λ

R ⁴ °\ R ¹

R ³

O R ³ O

146 147

211

Table 20.25

λ λ

^{^} w ³ λ

^' R ^{5 '} R ⁴

148 149 150

λ

^{^} R ¹ λ λ

^' H ^{^} R ³

151 152 153

(< λV ³ λ .R ⁵ r< R ⁴

154 155 156

<< .R ¹ ^ H << .R ³

157 158 159

212

Table 20.26

^\ JN ³ <<. ,R5 λ *4

H H H

160 161 162

λ .R ¹ << -H λ ,R ³

^{H H H}

163 164 165

^ .W ³ ^ ^R ⁵ ^N. _^ R ⁴

R ³ R ³ R ³

166 167 168

^\ / ^Rl ^ -H ^\ _^ R ³

R ³ _{3 R} 3

169 170 171

213

Table 20.27

172 173

O

<< .R ¹ 5a 'R ⁴

174 175

O

λ .R ⁵ *

^* σ

H

176 177

178

179

214

Xabi,

^Q ri O

\ .£ ⁵ *

O'

180

181

182

W

184

185

215

Table 20.29

O O

λ λ w ³ "Cf R ⁵

186 187

O O

A ti R ⁴ R ¹

188 189

O λ X) ^' CH ₃

190 191

O

λ CH ₃

\r "σ O I I C IH «3,

λ

192

193

216

Table 20.30

194

195

λ CH ₅

XT

196

197

198

O

199

217

5 Tabte2O31

O

O

λ .R 5a ,W ³

O ^' a^

λ .R 5a R ⁵ σ

200

201

O

O

λ ,R ^! 5a R ⁴

O R ¹

202

203

O O

λ _? 5a -H λ ,R ^! 5a ct^ σ

204 205 HA

O

O J T _j 5a

λ ,R 5a ^{^} CH ₃

207

206

218

Tafel ^gj fi .32 CH ₃

O

λ R ^: 5a O CH ₃

^

5a

λ R ^: O -a%

/

208 O

209 o ct^

.CH ₃

λ R 5 ^: a Cft xS

O

CH ₃

210

λ R 5 ^: a

O

-c^ σ%

211

213

219

Table 20.33

O

A -W ³ O o^

A R ⁵ O^

214

215

O

A R ⁴ xS O

216

217

O

218

221

220

220

Tak! _§ L2&2£ CH ₃

O

CH ₃

λ O

λ CH ₃

222

223 o CH ₃

.CH ₃

λ CH ₃

O

O

CH ₃

224

O ^'

CH ₃

225

O

O

227

221

Table 20.35

λ λ ,R ^; 5a Xk^ w ³ R ⁵

O O

228 229

λ o. λ 5a

Ό R ⁴ XT

O O

230 231

λ ,R ¹ 5a

- ^θ \ A R 5a λ^

W ³ R ⁵

H O H O

232 233

I^L R ^5a o> ^

^{^} R ⁴ °\ R ¹

H O H O

234 235

222

O.

-R ⁵

-M""

R ³ O

237

A O.

^■ R ¹

λ

^ °-^ ■ ^

R ^a O 239

R ³ O

238

λ R°

λ >< /

R- ¹ 241

240

λ

λ R^

243

242

223

Table 20.37

244 245

246 ^s^ 247

Pd ¹ and Pd ² of the "Sc" structures of Table 1.1 can also be independently selected from Table 30.1, below: Table 30.1

CH ₃

CH ₃

10

H ₃ C H,C

CH-j

OW

224

HsC

CH ₃

249

225

CH

/

253 254

255 256 257

10 Combinations of "Sc" and Pd ¹ and Pd ² independently selected from table

30.1 can be expressed in the form of ScPd ¹ .Pd ² , where Sc is represented by the respective letter A-G from Table 1.1 and Pd ¹ and Pd ² are represented by the respective number from Table 30.1. Thus, A.256.256 represents the following compound:

226

O ^

HO — jk ^CL n M

HO :χy

OH

Table 7: List of Compounds of MBF

A.254.67, A.254.68, A.254.69, A.254.78, A.254.79, A.254.80, A.254.248, A.254.249,

10 A.254.250, A.254.251. A.254.252, A.254.253, B.254.67, B.254.68, B.254.69, B.254.78, B.254.79, B.254.80, B .254.248, B.254.249, B.254.250, B.254.251, B.254.252, B.254.253, C.254.67, C.254.68, C.254.69, C.254.78, C.254.79, C.254.80, C.254.248, C.254.249, C.254.250, C.254.251, C.254.252, C.254.253, D.254.67, D.254.68, D.254.69, D.254.78, D.254.79, D.254.80, D.254.248, D.254.249, D.254.250, D.254.251, D.254.252,

15 D.254.253, E.254.67, E.254.68, E.254.69, E.254.78, E.254.79, E.254.80, E.254.248,

E.254.249, E.254.250, E.254.251, E.254.252, E.254.253, F.254.67, F.254.68, F.254.69, F.254.78, F.254.79, F.254.80, F.254.248, F.254.249, F.254.250, F.254.251, F.254.252, F.254.253, G.254.67, G.254.68, G.254.69, G.254.78, G.254.79, G.254.80, G.254.248, G.254.249, G.254.250, G.254.251, G.254.252, G.254.253, A.255.67, A.255.68, A.255.69,

20 A.255.78, A.255.79, A.255.80, A.255.248, A.255.249, A.255.250, A.255.251, A.255.252, A.255.253, B.255.67, B.255.68, B.255.69, B.255.78, B.255.79, B.255.80, B.255.248, B.255.249, B.255.250, B.255.251, B.255.252, B.255.253, C.255.67, C.255.68, C.255.69, C.255.78, C.255.79, C.255.80, C.255.248, C.255.249, C.255.250, C.255.251, C.255.252, C.255.253, D.255.67, D.255.68, D.255.69, D.255.78, D.255.79, D.255.80, D.255.248,

25 D.255.249, D.255.250, D.255.251, D.255.252, D.255.253, E.255.67, E.255.68, E.255.69, E.255.78, E.255.79, E.255.80, E.255.248, E.255.249, E.255.250, E.255.251, E.255.252, E.255.253, F.255.67, F.255.68, F.255.69, F.255.78, F.255.79, F.255.80, F.255.248, F.255.249, F.255.250, F.255.251, F.255.252, F.255.253, G.255.67, G.255.68, G.255.69, G.255.78, G.255.79, G.255.80, G.255.248, G.255.249, G.255.250, G.255.251, G.255.252,

30 G.255.253, A.67.67, A.68.68, A.69.69, A.78.78, A.79.79, A.80.80, A.248.248,

A.249.249, A.250.250, A.251.251, A252.252, A.253.253, B.67.67, B.68.68, B.69.69, B.78.78, B.79.79, B.80.80, B.248.248, B.249.249, B.250.250, B.251.251, B252.252, B.253.253, C.67.67, C.68.68, C.69.69, C.78.78, C.79.79, C.80.80, C.248.248, C.249.249,

227

5 C.250.250, C.251.251 , C252.252, C.253.253, D.67.67, D.68.68, D.69.69, D.78.78,

D.79.79, D.80.80, D.248.248, D.249.249, D.250.250, D.251.251, D252.252, D.253.253, E.67.67, E.68.68, E.69.69, E.78.78, E.79.79, E.80.80, E.248.248, E.249.249, E.250.250, E.251.251, E252.252, E.253.253, F.67.67, F.68.68, F.69.69, F.78.78, F.79.79, F.80.80, F.248.248, F.249.249, F.250.250, F.251.251, F252.252, F.253.253, G.67.67, G.68.68, 10 G.69.69, G.78.78, G.79.79, G.80.80, G.248.248, G.249.249, G.250.250, G.251.251, G252.252, G.253.253, A.256.257, B.256.257, C.256.257, D.256.257, E.256.257, F.256.257, G.256.257.

Methods of Inhibition of HCV polymerase

Another aspect of the invention relates to methods of inhibiting the

15 activity of HCV polymerase comprising the step of treating a sample suspected of containing HCV with a composition of the invention.

Compositions of the invention may act as inhibitors of HCV polymerase , as intermediates for such inhibitors or have other utilities as described below. The inhibitors will bind to locations on the surface or in a cavity of HCV

20 polymerase having a geometry unique to HCV polymerase . Compositions binding HCV polymerase may bind with varying degrees of reversibility. Those compounds binding substantially irreversibly are ideal candidates for use in this method of the invention. Once labeled, the substantially irreversibly binding compositions are useful as probes for the detection of HCV polymerase .

25 Accordingly, the invention relates to methods of detecting HCV polymerase in a sample suspected of containing HCV polymerase comprising the steps of: treating a sample suspected of containing HCV polymerase with a composition comprising a compound of the invention bound to a label; and observing the effect of the sample on the activity of the label. Suitable labels are well known in

30 the diagnostics field and include stable free radicals, fluorophores, radioisotopes, enzymes, chemiluminescent groups and chromogens. The compounds herein are labeled in conventional fashion using functional groups such as hydroxy 1,

228

5 ' carboxyl, sulfhydryl or amino.

Within the context of the invention, samples suspected of containing HCV polymerase include natural or man-made 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, 10 and the like); laboratory samples; food, water, or air samples; bioproduct samples such as extracts of cells, particularly recombinant cells synthesizing a desired glycoprotein; and the like. Typically the sample will be suspected of containing an organism which produces HCV polymerase , frequently a pathogenic organism such as HCV. Samples can be contained in any medium 15 including water and organic solvent \ water mixtures. Samples include living organisms such as humans, and man made materials such as cell cultures.

The treating step of the invention comprises adding the composition of the invention to the sample or it comprises adding a precursor of the composition to the sample. The addition step comprises any method of 20 administration as described above.

If desired, the activity of HCV polymerase after application of the . composition can be observed by any method including direct and indirect methods of detecting HCV polymerase activity. Quantitative, qualitative, and semiquantitative methods of determining HCV polymerase activity are all 25 contemplated. Typically one of the screening methods described above are applied, however, any other method such as observation of the physiological properties of a living organism are also applicable.

Organisms that contain HCV polymerase include the HCV virus. The compounds of this invention are useful in the treatment or prophylaxis of HCV 30 infections in animals or in man.

However, in screening compounds capable of inhibiting human immunodeficiency viruses, it should be kept in mind that the results of enzyme

229

5 assays may not correlate with cell culture assays. Thus, a cell based assay should be the primary screening tool.

Screens for HCV polymerase Inhibitors.

Compositions of the invention are screened for inhibitory activity against HCV polymerase by any of the conventional techniques for evaluating enzyme

10 activity. Within the context of the invention, typically compositions are first screened for inhibition of HCV polymerase in vitro and compositions showing inhibitory activity are then screened for activity in vivo. Compositions having in vitro Ki (inhibitory constants) of less then about 200 X 10" ⁶ M, typically less than about 100 X 10" ⁷ M and preferably less than about 50 X 10" ⁸ M are preferred for

15 in vivo use.

Useful in vitro screens have been described in detail and will not be elaborated here. However, the examples describe suitable in vitro assays.

The compounds of the present invention have HCV CC50 values (μM) in the range of about 0.1 to about 1000, or about 0.1 to about 500, or about 0.1 to

20 about 400, or about 0.1 to about 300, or about 0.1 to about 200, or about 0.1 to about 100, or less than about 500, or less than about 400, or less than about 300, or less than about 200, or less than about 100, or less than about 50, or less than about 20, or less than about 10.

The compounds of the present invention have HCV EC50 values (μM) in

25 the range of about 0.1 to about 500, or about 0.1 to about 400, or about 0.1 to about 300, or about 0.1 to about 200, or about 0.1 to about 100, or about 0.1 to about 50, or less than about 500, or less than about 400, or less than about 300, or less than about 200, or less than about 100, or less than about 50, or less than about 20, or less than about 10.

30 Salts and Hydrates

230

5 The compositions of this invention optionally comprise salts of the compounds herein, especially pharmaceutically acceptable non-toxic salts containing, for example, Na ⁺ , Li ⁺ , K ⁺ / Ca ⁺² and Mg ⁺² . Such salts may include those derived by combination of appropriate cations such as alkali and alkaline earth metal ions or ammonium and quaternary amino ions with an acid anion 10 moiety, typically a carboxylic acid. Monovalent salts are preferred if a water soluble salt is desired.

Metal salts typically are prepared by treating a metal hydroxide with a compound of this invention. Examples of metal salts which are prepared in this way are salts containing Li ⁺ , Na ⁺ , and K ⁺ . A less soluble metal salt may be 15 precipitated from the solution of a more soluble salt by addition of the suitable metal compound.

In addition, salts may be formed from acid addition of certain organic and inorganic acids, e.g., HCl, HBr, ^SO^ H3PO4 or organic sulfonic acids, to basic centers, typically amines, or to acidic groups. Finally, it is to be understood that 20 the compositions herein comprise compounds of the invention in their unionized, as well as zwitterionic form, and combinations with stoichiometric amounts of water as in hydrates.

Also included within the scope of this invention are the salts of the parental compounds with one or more amino acids. Any of the amino acids 25 described above are suitable, especially the naturally-occurring amino acids found as protein components, although the amino acid typically is one bearing a side chain with a basic or acidic group, e.g., lysine, arginine or glutamic acid, or a neutral group such as glycine, serine, threonine, alanine, isoleucine, or leucine.

231

5 Pharmaceutical Formulations

The compounds of this invention are formulated with conventional carriers and excipients, which will be selected in accord with ordinary practice. Tablets will contain excipients, glidants, fillers, binders and the like. Aqueous formulations are prepared in sterile form, and when intended for delivery by

10 other than oral administration generally will be isotonic. All formulations will optionally contain excipients such as those set forth in the Handbook of Pharmaceutical Excipients (1986), herein incorporated by reference in its entirety. Excipients include ascorbic acid and other antioxidants, chelating agents such as EDTA, carbohydrates such as dextrin, hydroxyalkylcellulose,

15 hydroxyalkylmethylcellulose, stearic acid and the like. The pH of the formulations ranges from about 3 to about 11, but is ordinarily about 7 to 10.

While it is possible for the active ingredients to be administered alone it may be preferable to present them as pharmaceutical formulations. The formulations of the invention, both for veterinary and for human use, comprise

20 at least one active ingredient, e.g. a compound of the present invention, together with one or more acceptable carriers and optionally other therapeutic ingredients. The carrier(s) must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and physiologically innocuous to the recipient thereof.

25 ^• The formulations include those suitable for the foregoing administration routes. The formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. Techniques and formulations generally are found in Remington's Pharmaceutical Sciences (Mack Publishing Co., Easton, Pa.), herein incorporated

30 by reference in its entirety. Such methods include the step of bringing into association the active ingredient with the carrier which constitutes one or more accessory ingredients. In general the formulations are prepared by uniformly

232

5 and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product.

Formulations of the present invention suitable for oral administration may be presented as discrete units such as capsules, cachets or tablets each

10 containing a predetermined amount of the active ingredient; as a powder or granules; as a solution or a suspension in an aqueous or non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion. The active ingredient may also be administered as a bolus, electuary or paste.

A tablet is made by compression or molding, optionally with one or more

15 accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the active ingredient in a free-flowing form such as a powder or granules, optionally mixed with a binder, lubricant, inert diluent, preservative, surface active or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered active ingredient moistened with an

20 inert liquid diluent. The tablets may optionally be coated or scored and optionally are formulated so as to provide slow or controlled release of the active ingredient.

For administration to the eye or other external tissues e.g., mouth and skin, the formulations are preferably applied as a topical ointment or cream

25 containing the active ingredient(s) in an amount of, for example, 0.075 to 20% w/w (including active ingredient(s) in a range between 0.1% and 20% in increments of 0.1% w/w such as 0.6% w/w, 0.7% w/w, etc.), preferably 0.2 to 15% w/w arid most preferably 0.5 to 10% w/w. When formulated in an ointment, the active ingredients may be employed with either a paraffinic or a water-miscible

30 ointment base. Alternatively, the active ingredients may be formulated in a cream with an oil-in-water cream base.

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5 If desired, the aqueous phase of the cream base may include, for example, at least 30% w/w of a polyhydric alcohol, i.e. an alcohol having two or more hydroxyl groups such as propylene glycol, butane 1,3-diol, mannitol, sorbitol, glycerol and polyethylene glycol (including PEG 400) and mixtures thereof. The topical formulations may desirably include a compound which enhances

10 absorption or penetration of the active ingredient through the skin or other affected areas. Examples of such dermal penetration enhancers include dimethyl sulphoxide and related analogs.

The oily phase of the emulsions of this invention may be constituted from known ingredients in a known manner. While the phase may comprise merely

15 an emulsifier (otherwise known as an emulgent), it desirably comprises a mixture of at least one emulsifier with a fat or an oil or with both a fat and an oil. Preferably, a hydrophilic emulsifier is included together with a lipophilic emulsifier which acts as a stabilizer. It is also preferred to include both an oil and a fat. Together, the emulsifier(s) with or without stabilizer(s) make up the so-

20 called emulsifying wax, and the wax together with the oil and fat make up the so-called emulsifying ointment base which forms the oily dispersed phase of the cream formulations.

Emulgents and emulsion stabilizers suitable for use in the formulation of the invention include Tween® 60, Span® 80, cetostearyl alcohol, benzyl alcohol,

25 myristyl alcohol, glyceryl mono-stearate and sodium lauryl sulfate.

The choice of suitable oils or fats for the formulation is based on achieving the desired cosmetic properties. The cream should preferably be a non-greasy, non-staining and washable product with suitable consistency to avoid leakage from tubes or other containers. Straight or branched chain, mono- or dibasic

30 alkyl esters such as di-isoadipate, isocetyl stearate, propylene glycol diester of coconut fatty acids, isopropyl myristate, decyl oleate, isopropyl palmitate, butyl stearate, 2-ethylhexyl palmitate or a blend of branched chain esters known as

234

5 Crodamol CAP may be used, the last three being preferred esters. These may be used alone or in combination depending on the properties required. Alternatively, high melting point lipids such as white soft paraffin and/or liquid paraffin or other mineral oils are used.

Pharmaceutical formulations according to the present invention comprise

10 one or more compounds of the invention together with one or more pharmaceutically acceptable carriers or excipients and optionally other therapeutic agents. Pharmaceutical formulations containing the active ingredient may be in any form suitable for the intended method of administration. When used for oral use for example, tablets, troches, lozenges,

15 aqueous or oil suspensions, dispersible powders or granules, emulsions, hard or soft capsules, syrups or elixirs may be prepared. Compositions intended for oral use may be prepared according to any method known to the art for the manufacture of pharmaceutical compositions and such compositions may contain one or more agents including sweetening agents, flavoring agents,

20 coloring agents and preserving agents, in order to provide a palatable preparation. Tablets containing the active ingredient in admixture with nontoxic pharmaceutically acceptable excipient which are suitable for manufacture of tablets are acceptable. These excipients may be, for example, inert diluents, such as calcium or sodium carbonate, lactose, lactose monohydrate,

25 croscarmellose sodium, povidone, calcium or sodium phosphate; granulating and disintegrating agents, such as maize starch, or alginic acid; binding agents, such as cellulose, microcrystalline cellulose, starch, gelatin or acacia; and lubricating agents, such as magnesium stearate, stearic acid or talc. Tablets may be uncoated or may be coated by known techniques including

30 microencapsulation to delay disintegration and adsorption in the gastrointestinal tract and thereby provide a sustained action over a longer period.

235

5 For example, a time delay material such as glyceryl monostearate or glyceryl distearate alone or with a wax may be employed.

Formulations for oral use may be also presented as hard gelatin capsules where the active ingredient is mixed with an inert solid diluent, for example calcium phosphate or kaolin, or as soft gelatin capsules wherein the active

10 ingredient is mixed with water or an oil medium, such as peanut oil, liquid paraffin or olive oil.

Aqueous suspensions of the invention contain the active materials in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients include a suspending agent, such as sodium

15 carboxymethylcellulose, methylcellulose, hydroxypropyl methylcelluose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia, and dispersing or wetting agents such as a naturally occurring phosphatide (e.g., lecithin), a condensation product of an alkylene oxide with a fatty acid (e.g., polyoxyethylene stearate), a condensation product of ethylene oxide with a long

20 chain aliphatic alcohol (e.g., heptadecaethyleneoxycetanol), a condensation product of ethylene oxide with a partial ester derived from a fatty acid and a hexitol anhydride (e.g., polyoxyethylene sorbitan monooleate). The aqueous suspension may also contain one or more preservatives such as ethyl or n-propyl p-hydroxy-benzoate, one or more coloring agents, one or more flavoring agents

25 and one or more sweetening agents, such as sucrose or saccharin.

Oil suspensions may be formulated by suspending the active ingredient in a vegetable oil, such as arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. The oral suspensions may contain a thickening agent, such as beeswax, hard paraffin or ceryl alcohol. Sweetening

30 agents, such as those set forth herein, and flavoring agents may be added to provide a palatable oral preparation. These compositions may be preserved by the addition of an antioxidant such as ascorbic acid.

236

5 Dispersible powders and granules of the invention suitable for preparation of an aqueous suspension by the addition of water provide 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 exemplified by those disclosed above. Additional

10 excipients, for example sweetening, flavoring and coloring agents, may also be present.

The pharmaceutical compositions of the invention may also be in the form of oil-in-water emulsions. The oily phase may be a vegetable oil, such as olive oil or arachis oil, a mineral oil, such as liquid paraffin, or a mixture of these.

15 Suitable emulsifying agents include naturally-occurring gums, such as gum acacia and gum tragacanth, naturally occurring phosphatides, such as soybean lecithin, esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan monooleate, and condensation products of these partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate. The

20 emulsion may also contain sweetening and flavoring agents. Syrups and elixirs may be formulated with sweetening agents, such as glycerol, sorbitol or sucrose. Such formulations may also contain a demulcent, a preservative, a flavoring or a coloring agent.

The pharmaceutical compositions of the invention may be in the form of a

25 sterile injectable preparation, such as a sterile injectable aqueous or oleaginous suspension. This suspension may be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents which have been mentioned herein. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable

30 diluent or solvent, such as a solution in 1,3-butane-diol or prepared as a lyophilized powder. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution. In

237

5 addition, sterile fixed oils may conventionally be employed as a solvent or suspending medium. For this purpose any bland fixed oil may be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid may likewise be used in the preparation of injectables.

The amount of active ingredient that may be combined with the carrier 10 material to produce a single dosage form will vary depending upon the host treated and the particular mode of administration. For example, a time-release formulation intended for oral administration to humans may contain approximately 1 to 1000 mg of active material compounded with an appropriate and convenient amount of carrier material which may vary from about 5 to 15 about 95% of the total compositions (weight: weight). The pharmaceutical composition can be prepared to provide easily measurable amounts for administration. For example, an aqueous solution intended for intravenous infusion may contain from about 3 to 500 μg of the active ingredient per milliliter of solution in order that infusion of a suitable volume at a rate of about 20 30 mL/hr can occur.

Formulations suitable for administration to the eye include eye drops wherein the active ingredient is dissolved or suspended in a suitable carrier, especially an aqueous solvent for the active ingredient. The active ingredient is preferably present in such formulations in a concentration of 0.5 to 20%, 25 advantageously 0.5 to 10% particularly about 1.5% w/w.

Formulations suitable for topical administration in the mouth include lozenges comprising the active ingredient in a flavored basis, usually sucrose and acacia or tragacanth; pastilles comprising the active ingredient in an inert basis such as gelatin and glycerin, or sucrose and acacia; and mouthwashes 30 comprising the active ingredient in a suitable liquid carrier.

Formulations for rectal administration may be presented as a suppository with a suitable base comprising for example cocoa butter or a salicylate.

238

5 Formulations suitable for intrapulmonary or nasal administration have a particle size for example in the range of 0.1 to 500 μm (including particle sizes in a range between 0.1 and 500 μm in increments such as 0.5 μm, 1 μm, 30 μm, 35 μm, etc.), which is administered by rapid inhalation through the nasal passage or by inhalation through the mouth so as to reach the alveolar sacs. Suitable

10 formulations include aqueous or oily solutions of the active ingredient. Formulations suitable for aerosol or dry powder administration may be prepared according to conventional methods and may be delivered with other therapeutic agents such as compounds heretofore used in the treatment or prophylaxis of infections as described herein.

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

Formulations suitable for parenteral administration include aqueous and

20 non-aqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of ^• the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents.

The formulations are presented in unit-dose or multi-dose containers, for

25 example sealed ampoules and vials, and may be stored in a freeze-dried

(lyophilized) condition requiring only the addition of the sterile liquid carrier, for example water for injection, immediately prior to use. Extemporaneous injection solutions and suspensions are prepared from sterile powders, granules and tablets of the kind previously described. Preferred unit dosage formulations

30 are those containing a daily dose or unit daily sub-dose, as herein above recited, or an appropriate fraction thereof, of the active ingredient.

239

5 It should be understood that in addition to the ingredients provided by the present invention the formulations of this invention may include other agents conventional in the art having regard to the type of formulation in question, for example those suitable for oral administration may include flavoring agents.

10 The invention further provides veterinary compositions comprising at least one active ingredient, e.g., a compound of the present invention together with a veterinary carrier.

Veterinary carriers are materials useful for the purpose of administering the composition and may be solid, liquid or gaseous materials which are

15 otherwise inert or acceptable in the veterinary art and are compatible with the active ingredient. These veterinary compositions may be administered orally, parenterally or by any other desired route.

Compounds of the invention can also be formulated to provide controlled release of the active ingredient to allow less frequent dosing or to improve the

20 pharmacokinetic or toxicity profile of the active ingredient. Accordingly, the invention also provided compositions comprising one or more compounds of the invention formulated for sustained or controlled release.

The effective dose of an active ingredient depends at least on the nature of the condition being treated, toxicity, whether the compound is being used

25 prophylactically (lower doses) or against an active disease or condition, the method of delivery, and the pharmaceutical formulation, and will be determined by the clinician using conventional dose escalation studies. The effective dose can be expected to be from about 0.0001 to about 100 rήg/kg body weight per day. Typically, from about 0.01 to about 10 mg/kg body weight per day. More

30 typically, from about 0.01 to about 5 mg/kg body weight per day. More typically, from about 0.05 to about 0.5 mg/kg body weight per day. For example, the daily candidate dose for an adult human of approximately 70 kg body weight will

240

5 range from 1 mg to 1000 mg, or between 5 mg and 500 mg, and may take the form of single or multiple doses.

In another embodiment, the present application provides pharmaceutical compositions comprising a compound of the present invention, or a pharmaceutically acceptable salt, solvate, and/or ester thereof, and a 10 pharmaceutically acceptable carrier or exipient.

Routes of Administration

One or more compounds of the invention (herein referred to as the active ingredients) are administered by any route appropriate to the condition to be

15 treated. 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 will be appreciated that the preferred route may vary with for example the condition of the recipient. An advantage of the compounds of this invention is that they are

20 orally bioavailable and can be dosed orally.

Combination Therapy

In one embodiment, the compounds of the present invention can be used alone, e.g., for inhibiting cytochrome P450 monooxygenase. In another

25 embodiment, the compounds of the present invention are used in combination with other active therapeutic ingredients or agents. Preferably, the other active therapeutic ingredients or agents are interferons, ribavirin analogs, NS3 protease inhibitors, alpha-glucosidase 1 inhibitors, hepatoprotectants, non-nucleoside inhibitors of HCV, and other drugs for treating HCV.

30 Combinations of the compounds of Formula I-IV are typically selected based on the condition to be treated, cross-reactivities of ingredients and pharmaco-properties of the combination. For example, when treating an

241

5 infection (e.g., HCV), the compositions of the invention are combined with other active therapeutic agents (such as those described herein).

Suitable active therapeutic agents or ingredients which can be combined with the compounds of Formula I-IV can include interferons, e.g., pegylated rIFN-alpha 2b, pegylated rIFN-alpha 2a, rIFN-alpha 2b, IFN alpha-2b XL, rlFN- 10 alpha 2a, consensus IFN alpha, infergen, rebif, locteron, AVI-005, PEG-infergen, pegylated IFN-beta, oral interferon alpha, feron, reaferon, intermax alpha, r-IFN- beta, infergen + actimmune, IFN-omega with DUROS, and albuferon; ribavirin analogs, e.g., rebetol, copegus, VX-497, and viramidine (taribavirin); NS5a inhibitors, e.g., A-831 and A-689; NS5b polymerase inhibitors, e.g., NM-283, 15 valopicitabine, R1626, PSI-6130 (R1656), HCV-796, BILB 1941, MK-0608, NM-107, R7128, VCH-759, PF-868554, GSK625433, and XTL-2125; NS3 protease inhibitors, e.g., SCH-503034 (SCH-7), VX-950 (Telaprevir), ITMN-191, and BILN-2065; alpha-glucosidase 1 inhibitors, e.g., MX-3253 (celgosivir) and UT-231B; hepatoprotectants, e.g., IDN-6556, ME 3738, MitoQ, and LB-84451; non- 20 nucleoside inhibitors of HCV, e.g., benzimidazole derivatives, benzo-1,2,4- thiadiazine derivatives, and phenylalanine derivatives; and other drugs for treating HCV, e.g., zadaxin, nitazoxanide (alinea), BIVN-401 (virostat), DEBIO- 025, VGX-410C, EMZ-702, AVI 4065, bavituximab, oglufanide, PYN-17, KPE02003002, actilon (CPG-10101), KRN-7000, civacir, GI-5005, ANA-975, XTL- 25 6865, ANA 971, NOV-205, tarvacin, EHC-18, and NIM811.

In yet another embodiment, the present application discloses pharmaceutical compositions comprising 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 30 pharmaceutically acceptable carrier or exipient.

According to the present invention, the therapeutic agent used in combination with the compound of the present invention can be any agent

242

5 having a therapeutic effect when used in combination with the compound of the present invention. For example, the therapeutic agent used in combination with the compound of the present invention can be interferons, ribavirin analogs, NS3 protease inhibitors, alpha-glucosidase 1 inhibitors, hepatoprotectants, non- nucleoside inhibitors of HCV, and other drugs for treating HCV.

10 In another embodiment, the present application provides pharmaceutical compositions comprising 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 selected from the group consisting of pegylated rIFN-alpha 2b, pegylated rIFN-alpha 2a, rIFN-alpha 2b, IFN alpha-

15 2b XL, rIFN-alpha 2a, consensus IFN alpha, infergen, rebif, locteron, AVI-005, PEG-infergen, pegylated IFN-beta, oral interferon alpha, feron, reaferon, intermax alpha, r-IFN-beta, infergen + actimmune, IFN-omega with DUROS, albuferon, rebetol, copegus, VX-497, viramidine (taribavirin), A-831, A-689, NM- 283, valopicitabine, R1626, PSI-6130 (R1656), HCV-796, BILB 1941, MK-0608,

20 NM-107, R7128, VCH-759, PF-868554, GSK625433/ XTL-2125, SCH-503034 (SCH- 7), VX-950 (Telaprevir), ITMN-191, and BILN-2065, MX-3253 (celgosivir), UT- 231B, IDN-6556, ME 3738, MitoQ, and LB-84451, benzimidazole derivatives, benzo-l,2,4-thiadiazine derivatives, and phenylalanine derivatives, zadaxin, nitazoxanide (alinea), BIVN-401 (virostat), DEBIO-025, VGX-410C, EMZ-702,

25 AVI 4065, bavituximab, oglufanide, PYN-17, KPE02003002, actilon (CPG-10101), KRN-7000, civacir, GI-5005, ANA-975, XTL-6865, ANA 971, NOV-205, tarvacin, EHC-18, and NIM811 and a pharmaceutically acceptable carrier or exipient.

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

30 a) a first pharmaceutical composition comprising a compound of the present invention, or a pharmaceutically acceptable salt, solvate, or ester thereof; and

243

5 . b) a second pharmaceutical composition comprising at least one additional therapeutic agent selected from the group consisting of HIV protease inhibiting compounds, HIV non-nucleoside inhibitors of reverse transcriptase, HTV nucleoside inhibitors of reverse transcriptase, HIV nucleotide inhibitors of reverse transcriptase, HIV integrase inhibitors, gp41 inhibitors, CXCR4

10 inhibitors, gpl20 inhibitors, CCR5 inhibitors, interferons, ribavirin analogs, NS3 protease inhibitors, alpha-glucosidase 1 inhibitors, hepatoprotectants, non- nucleoside inhibitors of HCV, and other drugs for treating HCV, and combinations thereof.

Combinations of the compounds of Formula I-IV and additional active

15 therapeutic agents may be selected to treat patients infected with HCV and other conditions such as HIV infections. Accordingly, the compounds of Formula I-IV may be combined with one or more compounds useful in treating HIV, for example HIV protease inhibiting compounds, HIV non-nucleoside inhibitors of reverse transcriptase, HTV nucleoside inhibitors of reverse transcriptase, HIV

20 nucleotide inhibitors of reverse transcriptase, HIV integrase inhibitors, gp41 inhibitors, CXCR4 inhibitors, gpl20 inhibitors, CCR5 inhibitors, interferons, ribavirin analogs, NS3 protease inhibitors, alpha-glucosidase 1 inhibitors, hepatoprotectants, non-nucleoside inhibitors of HCV, and other drugs for treating HCV.

25 More specifically, 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, e.g., amprenavir, atazanavir, fosamprenavir, indinavir, lopinavir, ritonavir, lopinavir + ritonavir, nelfinavir, saquinavir, tipranavir, brecanavir, darunavir, TMC-126, TMC-114, mozenavir (DMP-450),

30 JE-2147 (AG1776), AG1859, DG35, L-756423, RO0334649, KNI-272, DPC-681,

DPC-684, and GW640385X, DG17, PPL-100, 2) a HIV non-nucleoside inhibitor of reverse transcriptase, e.g., capravirine, emivirine, delaviridine, efavirenz,

244

5 nevirapine, (+) calanolide A ₇ etravirine, GW5634, DPC-083, DPC-961, DPC-963, MIV-150, and TMC-120, TMC-278 (rilpivirine), efavirenz, BILR 355 BS ₇ VRX 840773, UK-453,061, RDEA806, 3) a HIV nucleoside inhibitor of reverse transcriptase, e.g., zidovudine, emtricitabine, didanosine, stavudine, zalcitabine, lamivudine, abacavir, amdoxovir, elvucitabine, alovudine, MIV-210, racivir (±-

10 FTC), D-d4FC, emtricitabine, phosphazide, fozivudine tidoxil, fosalvudine tidoxil, apricitibine (AVX754), amdoxovir, KP-1461, abacavir + lamivudine, abacavir + lamivudine + zidovudine, zidovudine + lamivudine, 4) a HTV nucleotide inhibitor of reverse transcriptase, e.g., tenofovir, tenofovir disoproxil fumarate + emtricitabine, tenofovir disoproxil fumarate + emtricitabine +

15 efavirenz, and adefovir, 5) a HIV integrase inhibitor, e.g., curcumin, derivatives of curcumin, chicoric acid, derivatives of chicoric acid, 3,5-dicaffeoylquinic acid, derivatives of 3,5-dicaffeoylquinic acid, aurintricarboxylic acid, derivatives of aurintricarboxylic acid, caffeic acid phenethyl ester, derivatives of caffeic acid phenethyl ester, tyrphostin, derivatives of tyrphostin, quercetin, derivatives of

20 quercetin, S-1360, zintevir (AR-177), L-870812, and L-870810, MK-0518

(raltegravir), BMS-707035, MK-2048, BA-011, BMS-538158, GSK364735C, 6) a gp41 inhibitor, e.g., enfuvirtide, sifuvirtide, FB006M, TRI-1144, SPC3, DES6, Locus gp41, CovX, and REP 9, 7) a CXCR4 inhibitor, e.g., AMD-070, 8) an entry inhibitor, e.g., SPOlA, TNX-355, 9) a gpl20 inhibitor, e.g., BMS-488043 and

25 BlockAide/CR, 10) a G6PD and NADH-oxidase inhibitor, e.g., immunitin, 10) a CCR5 inhibitor, e.g., aplaviroc, vicriviroc, INCB9471, PRO-140, INCB15050, PF- 232798, CCR5mAb004, and maraviroc, 11) an interferon, e.g., pegylated rlFN- alpha 2b, pegylated rIFN-alpha 2a, rIFN-alpha 2b, IFN alpha-2b XL, rIFN-alpha 2a, consensus IFN alpha, infergen, rebif, locteron, AVI-005, PEG-infergen,

30 pegylated IFN-beta, oral interferon alpha, feron, reaferon, intermax alpha, r-IFN- beta, infergen + actimmune, IFN-omega with DUROS, and albuferon, 12) ribavirin analogs, e.g., rebetol, copegus, VX-497, and viramidine (taribavirin) 13)

245

5 NS5a inhibitors, e.g., A-831 and A-689, 14) NS5b polymerase inhibitors, e.g.,

NM-283, valopicitabine, R1626, PSI-6130 (R1656), HCV-796, BILB 1941, MK-0608, NM-107, R7128, VCH-759, PF-868554, GSK625433, and XTL-2125, 15) NS3 protease inhibitors, e.g., SCH-503034 (SCH-7), VX-950 (Telaprevir), ITMN-191, and BILN-2065, 16) alpha-glucosidase 1 inhibitors, e.g., MX-3253 (celgosivir) and

10 UT-231B, 17) hepatoprotectants, e.g., IDN-6556, ME 3738, MitoQ, and LB-84451, 18) non-nucleoside inhibitors of HCV, e.g., benzimidazole derivatives, benzo- 1,2,4- thiadiazine derivatives, and phenylalanine derivatives, 19) other drugs for treating HCV, e.g., zadaxin, nitazoxanide (alinea), BIVN-401 (virostat), DEBIO- 025, VGX-410C, EMZ-702, AVI 4065, bavituximab, oglufanide, PYN-17,

15 KPE02003002, actilon (CPG-10101), KRN-7000, civacir, GI-5005, ANA-975, XTL- 6865, ANA 971, NOV-205, tarvacin, EHC-18, and NIM811, 19) pharmacokinetic enhancers, e.g., BAS-100 and SPI452, 20)RNAse H inhibitors, e.g., ODN-93 and ODN-112, 21) other anti-HIV agents, e.g., VGV-I, PA-457 (bevirimat), ampligen, HRG214, cytolin, polymun, VGX-410, KD247, AMZ 0026, CYT 99007, A-221 HIV,

20 BAY 50-4798, MDXOlO (iplimumab), PBSl 19, ALG889, and PA-1050040.

It is also possible to combine any compound of the invention with one or more other active therapeutic agents in a unitary dosage form for simultaneous or sequential administration to a patient. The combination therapy may be administered as a simultaneous or sequential regimen. When administered

25 sequentially, the combination may be administered in two or more administrations.

Co-administration of a compound of the invention with one or more other active therapeutic agents generally refers to simultaneous or sequential administration of a compound of the invention and one or more other active

30 therapeutic agents, such that therapeutically effective amounts of the compound of the invention and one or more other active therapeutic agents are both present in the body of the patient.

246

5 Co-administration includes administration of unit dosages of the compounds of the invention before or after administration of unit dosages of one or more other active therapeutic agents, for example, administration of the compounds of the invention within seconds, minutes, or hours of the administration of one or more other active therapeutic agents. For example, a

10 unit dose of a compound of the invention can be administered first, followed within seconds or minutes by administration of a unit dose of one or more other active therapeutic agents. Alternatively, a unit dose of one or more other therapeutic agents can be administered first, followed by administration of a unit dose of a compound of the invention within seconds or minutes. In some

15 cases, it may be desirable to administer a unit dose of a compound of the invention first, followed, after a period of hours (e.g., 1-12 hours), by administration of a unit dose of one or more other active therapeutic agents. In other cases, it may be desirable to administer a unit dose of one or more other active therapeutic agents first, followed, after a period of hours (e.g., 1-12 hours),

20 by administration of a unit dose of a compound of the invention.

The combination therapy may provide "synergy" and "synergistic effect", i.e. the effect achieved when the active ingredients used together is greater than the sum of the effects that results from using the compounds separately. A synergistic effect may be attained when the active ingredients are: (1) co-

25 formulated and administered or delivered simultaneously in a combined formulation; (2) delivered by alternation or in parallel as separate formulations; or (3) by some other regimen. When delivered in alternation therapy, a synergistic effect may be attained when the compounds are administered or delivered sequentially, e.g., in separate tablets, pills or capsules, or by different

30 injections in separate syringes. In general, during alternation therapy, an effective dosage of each active ingredient is administered sequentially, i.e.

247

5 serially, whereas in combination therapy, effective ^" dosages of two or more active ingredients are administered together.

In still yet another embodiment, the present application provides for methods of inhibiting HCV polymerase in a cell, comprising: contacting a cell infected with HCV with an effective amount of a compound of Formula I-IV, or

10 a pharmaceutically acceptable salt, solvate, and/or ester thereof, whereby HCV polymerase is inhibited.

In still yet another embodiment, the present application provides for methods of inhibiting HCV polymerase in a cell, comprising: contacting a cell infected with HCV with an effective amount of a compound of Formula I-IV, or

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

20 a pharmaceutically acceptable salt, solvate, and/or ester thereof, and at least one additional active therapeutic agent selected from the group consisting of interferons, ribavirin analogs, NS3 protease inhibitors, alpha-glucosidase 1 inhibitors, hepatoprotectants, non-nucleoside inhibitors of HCV, and other drugs for treating HCV.

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

In still yet another embodiment, the present application provides for

30 methods of treating HCV in a patient, comprising: administering to the patient a therapeutically effective amount of a compound of Formula I-IV, or a

248

5 pharmaceutically acceptable salt, solvate, and/or ester thereof, and at least one additional active therapeutic agent, whereby HCV polymerase is inhibited. In still yet another embodiment, the present application provides for methods of treating HCV in a patient, comprising: administering to the patient a therapeutically effective amount of a compound of Formula I-IV, or a

10 pharmaceutically acceptable salt, solvate, and/or ester thereof, and at least one additional active therapeutic agent selected from the group consisting of interferons, ribavirin analogs, NS3 protease inhibitors, alpha-glucosidase 1 inhibitors, hepatoprotectants, non-nucleoside inhibitors of HCV, and other drugs for treating HCV.

15 In still yet another embodiment, the present application provides for the 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 treating an HCV infection in a patient.

20 Examples

Exemplary methods for the preparation of the compounds of the invention are provided below. These methods are intended to illustrate the nature of such preparations and are not intended to limit the scope of applicable

25 methods. While the examples specify certain reaction conditions, one skilled in the art will understand how to vary the specific reaction conditions to obtain the full scope of the invention. Exemplary methods for the preparation of the compounds of the invention are illustrated in Schemes 2, 3, 5, 6, 7, 8, 9, 10, 30, 31, and 32.

30 Certain abbreviations and acronyms are used in describing the .

. experimental details. Although most of these would be understood by one

249

5 skilled in the art, Table IE contains a list of many of these abbreviations and acronyms.

250

10

15

251

Example 1

Scheme 2

NH ₂ H ₂ N^% o Il Persilylated

'?- 8-aza adenine O

^Sa ' ⁰ ^Vx- ⁰ yV. .OAc Il N -\M KCi

EtO

O i^Et°γ°y' Eto'i^- ⁰ v^ ⁰ y ^N

CH ₃ CN. BF ₃ Et ₂ O, OEt OBz 45 ⁰ C, 2 h

OBz OBz

1.7 2.1 2.2

NH,

TMS-Br O N' ^' Tl "7 >• cαπc. INn ₄ UH _{1 Q} JV^N Ala-O-Bu..HCI, TEA effort »θiy ^θ yy«^ ii.cV ^" HPLC HO i^-' V V* ^N Py- ^{60 c} .4h; 25 %

VJ

OBz OH 2.3 2.4

NH,

"BuO. N^T .J

J HNH X_7

OH ⁿ BuO

2.5

.N- .N-

\ ^V _N I. cone. NH ₄ OH, ^HaN ^f \ Ala-0-Bu..HCI.TEA

O >Y 45 °C _. ^/ Ph ₃ P, Aldrithiol ^P Il^O^ _O > _' N "-C18-HPLC

HO ^'

OH xy S o _n VN ^py - ⁶⁰ ° ^c - ^4h : ^25%

OBz ^Ho i^V/

OH

2.6 2.7

H ₂ N-^ ^N" ^\

^" ^Y S^N H^NH V \yV ^' *

T ^ OH

"BuO

2.8

10

252

5 Benzoic acid 2-(7-amino-[l,2,3]triazolo[4,5-d]pyiliiiidin-3-yI)-5-(dietho xy- phosphoiylmethoxy)-tetrahydrofyian-3-yl ester, (2.1) and benzoic acid 2-(7- ainino-[l,2,3]triazolo[4,5-d]pyrintidin-2-yl)-5-(diethoxy-ph osphorylinethoxy)- tetrahydrofyran-3-yl ester (2.2)

10 A 1,1,1,3,3,3-hexarnethyldisilazane suspension (10 mL) of 8-azaadenine (0.25 g, 1.8 mmol) and ammonium sulfate (0.16 g, 1.2 mmol) was heated at 140 ⁰ C for 1.5 h. The resulting clear solution was concentrated to dryness and azeotroped with acetonitrile twice. The solution of the persilylated 8-azaadenine was added to phosphonate 1.7 (see scheme 1) (0.5 g, 1.2 mmol) in acetonitrile (10 mL), and the

15 resultant solution then treated with BF3.Et2θ (0.24 mL, 1.8 mmol). The resulting solution was heated at 45 °C for 1.5 h and then concentrated to dryness. The crude residue was purified by C-18 HPLC to afford the title compound 2.1 (0.16 g/37 %) and 2.2 (0.07 g, 12 %). Compound 2.1 ¹ H NMR (CDCIs): δ 8.45 (s, IH), 8.00 (d, 2H, J= 8.0), 7.56 (t, IH, J = 7.3), 7.42 (t, 2H, J = 7.6), 6.68 (d, IH, J= 1.9),

20 6.49-6.54 (m, IH), 5.60-5.70 (m, IH), 4.01-4.18 (m, 4H), 3.64-3.80 (m, 2H), 3.08-3.17 (m, IH), 2.64-2.72 (m, IH), 1.20-1.34 (2t,6H). ³¹ P NMR: 20.55 ppm. Compound 2.2 ¹ H NMR (CDCl ₃ ): δ 8.48(s, IH), 8.00 (d, 2H, J = 8.0), 7.59 (t, IH, J = 7.6), 7.42 (t, 2H, J = 7.6), 6.65 (d, IH, J= 3.3), 6.15-6.21 (m, IH), 5.55 (m, lH), 4.04-4.22 (m, 5H), 3.69 (t, IH, J = 2.4),.2.94-3.01 (m, IH), 2.49-2.60 (m, IH), 1.20-1.34 (2t, 6H). ³¹ P NMR:

25 20.68 ppm.

[5-(7-Amino-[l,2,3]triazolo[4,5-d]pyriinidin-3-yl)-4-hydr oxy-teixahydrofuran-2- yloxymethyl]-phosphonic acid (2.4)

30 A solution of compound 2.1 (300 mg, 0.6 mmol) in acetonitrile (5 mL) was treated with bromotrimethylsilane (0.93 g, 6 mmol) and 2,6-lutidine at room temperature for 3.5 h. The mixture was concentrated to dryness and azeotroped with cone. NH4OH twice. The residue was subjected to C-18 HPLC eluting with to give 180 mg (68 % yield) diacid 2.3. The diaάd 2.3 was then treated with

35 concentrated NH4OH (10 ml) at 45 ⁰ C for 30 min. The reaction mixture was

253

5 concentrated to dryness and purified by C-18 HPLC to afford compound 2.4 (94 mg, 68 %). ¹ H NMR P2O): δ 8.25 (s, 1H),6.26 (d, IH, /= 2.4), 5.42-5.52 (m, 2H), 3.32 (d, 2H, / = 9.1), 2.54-2.61 (m, IH), 2.33-2.42 (m, IH); ³¹ P NMR: 15.39 ppm.

[5-(7-Amino-[l,2,3]iaiazolo[4 _/ 5-d]p3τimidin-2-yl)-4-hydroxy-tetrahydroftu-aϋn 10 yloxymethyll-phosphonic acid (2.7)

Compound 2.7 (76 mg, 71 %) was obtained from 2.2 using the procedure described for the preparation of 2.4. ¹ H NMR (D ₂ O): δ 8.21 (s, 1H),6.25 (d, IH, / = 2.4), 563 (t, IH, J= 4.0), 4.99-5.04 (m, IH), 3.48-3.56 (m, IH), 3.32-3.40 (m, IH), 15 2.46-2.54 (m, IH), 2.28-2.37 (m, IH); ³¹ P NMR: 13.03 ppm. LRMS (ESI) MH ⁺ C&H^NeOeP requires 333.1. Found 332.9.

Bis-amidate prodrug 2.5: Phosphonic acid 2.4 (30 mg, 0.09 mmol) was dissolved in pyridine (0.5 mL) and treated with (S)-Alanine n-butyl ester hydrochloride (98 mg, 0.54 mmol) and TEA (91 mg, 0.90 mmol). A freshly prepared solution of

20 PhsP (165 mg, 0.63 mmol) and 2-Aldrithiol (140 mg, 0.66 mmol) in pyridine (1 mL) was added. The resulting mixture was heated at 65°C for 4 h and then concentrated to dryness. The crude resiude was purified on silica gel column chromatography (eluting with EtOAc and then 30 % Ethanol /70 % EtOAc) and repurified on C-18 HPLC to afford 14.5 mg of prodrug 2.4. ¹ H NMR (CDsOD): δ

25 8.36 (s, IH), 6.31 (d, IH, / = 3.7), 5.45-5.48 (m, IH), 5.34-5.40 (m, IH), 3.92-4.20 (m, 6H), 3.75-3.82 (m, IH), 3.61-3.70 (m, IH), 2.64-2.72 (m, IH), 2.35-2.44 (m, IH), 1.53-1.70 (m, 4H), 1.30-1.47 (m, 10 H), 0.89-1.00 (m, 6H); ³¹ P NMR: 23.30 ppm. LRMS (ESI) MH ⁺ C^H ₃₉ N ₈ OeP requires 587.3. Found 587.2.

Diphosphophosphonate of 2.4: The diphosphophosphonate of 2.4 was 30 prepared as the same manner as the diphosphophosphonate of 5.2 as described below.

254

5 Bis-amidate prodrug 2.8: Prepared as described for the synthesis of 2.5. Yield 15 mg (26 %). ¹ H NMR (CD ₃ OD): δ 8.31 (s, IH), 6.27 (d, IH, / = 3.0), 5.52-5.55 (m, IH), 5.13-5.17 (m, IH), 3.92-4.20 (m, 7H), 3.69-3.77 (m, IH), 2.60-2.67 (m, IH), 2.32-2.40 (m, IH), 1.53-1.70 (m, 4H), 1.30-1.47 (m, 10 H), 0.89-0.98 (2t, 6H); ³¹ P NMR: 23.19 ppm.

10

Diphosphophosphonate of 2.7: The diphosphophosphonate of 2.7 was prepared as the same manner as the diphosphophosphonate of 5.2 as described below. ¹ H NMR (CD.OD): δ 2.33-2.38 (m, IH), 2.54-2.65 (m, IH), 3.58-3.70 (m, 2H), 5.20-5.30 (m, IH), 5.57-5.65 (m, IH), 6.00 (d, IH, / = 3.4). ³¹ PNMR: 8.11, -10.68,

15 -23.23 ppm.

20

25

30

255

Example 2

Scheme 3

NH, NHBz NHBz i. TBDPSCI, pyr,

<f Y V ^L DMTrCI, pyr yy Tj --NN D DMMAAPI

/ ^V/ VN^N^ " ^{• BZCI} - ^Pyf λ/V "N

> \_J ^{*" DMT} r0 \_J

OH OH δTBDPS 1 3

NBz ₂ NHBz i. mCPBA HO^PO ₃ Et ₂

' ^{• BzCI} - PVf ^OyN^^ 'B"OH , ^ ^ y

OTBDPS OTBDPS

NHBz NHBz ϊ i. TMSBr, A

^N-~^W _N i. DMSO, DCC ❖ Y^ ^N " ^{■ NH} 4° ^H

Et ₂ O ₃ Pv^-O, iλ..^ «■ ^H ₂ P ⁽ Ph ⁾ ₃ ^Ci . _{aojSk A} λ^I^ iii. TBAF nBuLi »- HO' iii. nBuLi / THF

OTBDPS OTBDPS

NH ₂ NH ₂

"BuQ

PPh ₃ Et ₃ N

H ₂ O ₃ Pv <ϊ Aldrithiol O >- H\N- °Pv

NH ₂ HN -°κ/\J "BUO

OH "BuO.

¹ Y^ V _N - OH

Lead Poisoned 5%Pd/BaCO ₃ Lead Poisoned 5%Pd/BaCO ₃ Quiπoliπe Quinoline

H ₂ (S) H ₈ (g)

NH ₂ NH ₂

N- "BuQ N IN--v^-5

H ₂ O ₃ Px -O O. xy 3 M HN- 9PV _N HN

OH ^nBuO γk yy - ^N -^w :

OH

10 11

10 Cordycepin 1 (3' deoxy adenosine) (Sigma Aldrich) is first 5'O protected by treatment with dimethoxy trityl chloride, and then N-6 benzoyl protected

256

5 according to the procedure of Charubala et al. (HeIv. Chim Acta 2002 p2284, herein incorporated by reference in its entirety), to give 2. Intermediate 2 is then 2'O protected by treatment with TBDMS chloride in the presence of pyridine and silver nitrate (see Tet Lett. 1981, p 4775, herein incorporated by reference in its entirety) and then the 5'O trityl group is removed by treatment with acetic

10 acid to give the intermediate 3. Treatment of 3 with triphenylphosphine and iodine according to the procedure of Maag et al. (J. Med. Chem. 1992, pl440 , herein incorporated by reference in its entirety) introduces the 5' iodine. Further treatment with benzoyl chloride then affords the bis benzoyl protected product 4 (Maag et al J. Med. Chem. 1992 pl440). Treatment of 4 with sodium methoxide

15 affords exocyclic alkene 5 (Maag et al J. Med. Chem. 1992, pl440). Treatment of alkene 5 with m-CPBA (m-chloroperbenzoic acid) in the presence of hydroxylmethyl diethylphosphonate afforded the alcohol 6 (see Maag et al J. Med. Chem. 1992 pl440). Alcohol 6 was then oxidized using Swern conditions, followed by Wittig olefination using (chloromethyl)triphenylphosphonium

20 chloride, and finally treatment with nBuLi at low temperature provided the alkyne 7 (see J. Med. Chem, 2004, p5041, Siddiqui et al.) both of which are herein incorporated by reference in their entirety. The alkyne 7 is then treated with TMSBr in acetonitrile to remove the phosphonate ester groups, followed by treatment with ammonium hydroxide to remove the benzoyl groups, and finally

25 TBAF to remove the 2'O-silyl group, thereby affording 8 (See scheme 1 and 4 and Greene and Wuts, protecting groups in Organic Chemistry, 3 ^rd Edition, Wiley, herein incorporated by reference in its entirety). Diacid 8 is then converted to the bis amidate prodrug 9 according to the procedure described in Scheme 1. Diacid 8 is then treated with lead poisoned-5% palladium on barium

30 carbonate in the presence of quinoline under about one to five atmospheres of hydrogen gas to give olefin 10. Similar treatment of bis amidate prodrug 9 will give olefin 11.

257

Example 3

Scheme 5

O

"NH

Persilylated I I) TMSBr

^« 4λCV .OAc 8-aza-G N NH ₂

CH ₃ CN, SnCI ₄ , ^Eto' L .O _v O _y N'

\_y 2) NH ₄ OH

OBz r.t., 1 day OBz

1.7 5.1

V CO ₂ Bu

O N ,«; ^N V Ji S J ^H NH ₂ HCI

HO-P _^ -. λ-; K' N- _T Jλl INH ^HNJ ;^ ^O -^°N^ ^{N N NH} 2

Aldrithiol-2, Ph ₃ P OH

OH Py, 65 ⁰ C

5.2 5.3

10

[5-(2-Ainino-6-oxo-l,6-dihydro-pιιrin-9-yl)-4-hydroxy-t etrahydro-ft-iaiι-2- yloxymethyl]-phosphonic acid (5.2).

8-aza-G was persilylated first using the procedure described for the preparation 15 of compound 2.2. Compound 1.7 (300 mg, 0.7 mmol) and the persilyated 8-aza-G base (1.8 mmol) were dissolved in 5 mL QHbCN, and treated with SnCk (IM solution in CH2CI2) (2.1 mL, 2.1 mmol). The mixture was stirred at r.t. for 24 h, quenched with NaHCCk (2 g) and water (3 mL), filtered, and concentrated down under reduced pressure. The residue was subjected to reverse phase HPLC 20 eluting with 5-95% CHsCN in water (0.1% TFA) to yield crude product 5.1 (45 mg, 14% yield). The crude product was carried on to the next two steps using the procedure described for the preparation of compound 2.4 to yield compound 5.2 (21 mg, 68% yield). ¹ HNMR (300 MHz, D2O) δ 2.35-2.41 (m, IH), 2.54-2.63 (m,

258

5 IH), 3.29-3.45 (m, 2H), 5.24-5.26 (m, IH), 5.58 (d, IH, / = 2.7), 6.04 (d, IH, / = 2.7) ³¹ PNMR : 24.32 ppm.

Bis-amidate prodrug 5.3.

Compound 5.3 (4.5 mg, 45% yield) was synthesized from compound 5.2 (7 mg,

10 0.02 mmol) using the procedure described for the preparation of compound 2.5. ¹ HNMR (300 MHz, CD ₃ OD) δ 0.92-1.00 (m, 6H), 1.36-1.45 (m, 10H), 1.60-1.70 (m, 4H), 2.30-2.40 (m, IH), 2.60-2.70 (m, IH), 3.65-3.70 (m, IH), 3.84-3.90 (m, IH), 3.90-4.20 (m, 6H), 5.25-5.28 (m, IH), 5.42 (s, IH), 6.09 (d, IH, J = 3.5). ³¹ PNMR : 24.32. LRMS (ESI) MH ⁺ C23H39N8O9P requires 603.3. Found 603.0.

15

Diphosphophosphonate of 5.2.

Compound 5.2 (6.0 mg, 0.0173 mmol) was dissolved in DMSO (.500 mL) and then treated with tributylamine (.021 mL, 0.087mmol) followed by carbonyldiimidazole (28 mg, 0.173mmol). The mixture was stirred at room

20 temperature for 1 h and then MeOH (.0063 mL, 0.156 mmol) was added. The mixture was stirred for an additional 30 min. Tributyl ammonium pyrophosphate (95 mg, 0.173 mmol) in DMF (0.4 mL) was added and the reaction mixture stirred for 1 h. The solvent was removed under reduced pressure and the crude product was purified by ion exchange HPLC (0-40%

25 TEAB) to provide the diphosphophosphonate (4.5 mg). ¹ HNMR (300 MHz, D2O) δ 1.11-1.20 (m, 27H), 2.38-2.45 (m, IH), 2.60-2.70 (m, IH), 2.85-3.25 (m, 18H), 3.40- 3.60 (m, 2H), 5.38-5.42 (m, IH), 5.58-5.61 (m, IH), 6.25 (d, IH, J = 2.8), 8.24 (s, IH). ³¹ PNMR: 8.25, -5.67, -21.46 ppm.

30

259

Example 4

Scheme 6

¹ PrO.? ,.OH ¹ PrC^

OBz

BzO ^{1 Br>} γ ^O N- ^> '--OBz i) HBr, AcOH 'PrO-Jl _x

¹ PrO ,O _* ^-°\..>.- 1) K ₂ CO ₃ , EtOH

-OBz

BzO OBz ii) SnCI ₄ , MeCN 2) TBDPSCI, Py F" OBZ Tetrahedron, 1998, 54 (28), 8223

O 1) BzCl, DMAP, pyr

¹ PrO-S n λX , ') Dess Martin _ip _ Tf _o 2) NH ₄ F. EtOH. iprO'^ H ii. NaBH ₄ 3) BAIB, TEMPO

OH OBz

NH ₂

¹ PrO . 8-aza adenine _lD

.0^ ¹ PrO 1 -Q" ⁰ N-J'^UoH Pb(OAc) ₄ & r^oJ^L _n

^oO^^/y°\^-OAc HMDS ^PrO - ³ _V ^O. ^{^} N^

F OBz DMF, Py P= _{OB2 BF3 θEt MeCN} ^{■ ■}

OBz

5 6

INπ ₂ ^• N

I) TMSBr N"

HO _^ o-^ λ ^w y-N N ^l 2) NaOEt HO :X ^{' ^} - ^{^} "- ii-Q »*

' ^o ~o- "Bu-Q HN

PPh ₃

F OH Aldrithiol Y< F OH

Et ₃ N 8

The commercially available benzoate 6.1 (CMS chemicals, UK) is 10 converted to the anomeric bromide according to the method described in J. Org Chem. 1985, p3644 (herein incorporated by reference in its entirety). The bromide is then reacted with the diisopropyl phosphonate in the presence of tin(IV) chloride in acetonitrile at reflux according to the procedure described in Tetrahedron, 1998, 54 (28), 8223, herein incorporated by reference in its entirety. 15 The product is an approximate 1:1 mixture of anomeric isomers that is carried forward as a mixture of isomers. The isomeric mixture 6.2 is then treated with potassium carbonate in methanol to provide the diol as a mixture of isomers.

260

5 After purification by silica gel chromatography the diol is then treated with TBDPS chloride in pyridine overnight to afford the 5'O-silyl protected mixture of isomers 6.3. At this stage the anomeric isomers are separated to give the desired beta-isomer 6.3. Protected intermediate 6.3 is then oxidized by treatment with 1.5 eq. of Dess Martine Periodinane reagent, and after work up

10 immediately reduced by dissolution in ethanol and treatment with sodium borohydride (6 eq.) at 0 C to afford 6.4. Compound 6.4 was then treated with benzoyl chloride in the presence of pyridine and DMAP (dimethylamino pyridine) to give the benzoate, which is then treated with pyridine.HF (10 eq.) in dichloromethane to provide the 5' OH intermediate. This intermediate is then

15 oxidized using BAIB and TEMPO (as in Scheme 1) to provide 6.5. Acid 6.5 is then treated with lead tetraacetate in DMF and pyridine overnight to afford the anomeric acetate 6.6 (as in Scheme 2). Acetate 6.6 is then reacted with a silylated nucleobase, for example silylated 8-azaadenine prepared from the treatment of 8-aza adenine with hexamethyldisilazide and ammonium sulfate, in the

20 presence of a Lewis acid e.g. BFsOEt to afford the nucleoside phosphonate 6.7. Phosphonate 6.7 is then treated with TMSBr (trimethylsilyl bromide) to generate the diacid, followed by sodium ethoxide to afford the desired diacid 6.8. Diacid 6.8 can be converted to the prodrug, e.g. the bis alanine n-butyl amidate 6.9, according to the method described in Scheme 2.

25 .. ;

Example 5 Scheme 7 O O _n Z=N

\-cJ M t)Bz \λ M O. Bz ^N i ^u

7.1 7.2

Benzoic acid 2-acetoxy-5-(diethoxy- 1-[3-Benzoyloxy-5-(diethoxy-phospriorylmethoxy)- phosphorylmethoxy)-tetrahydro-furan-3-yl ester tθtrahydro-furan-2-yl]-

" * * 1H-[1,2,4]triazole-3-carboxylic acid methyl ester

261

5 l-[3-Benzoyloxy-5-(diethoxy-phosphorylmethoxy)-tetrahydro-ft ιran-2-yl]- UEf-[l,2,4]triazole-3-carboxylic acid methyl ester.

Benzoic acid 2-acetoxy-5-(diethoxy-phosphorylmethoxy)-tetrahydro-furcin-3 -yl ester (170 mg, 0.4 mmol), lH-tlA^Triazole-S-carboxylic acid methyl ester (51

10 mg, 0.4 mmol) and Bis-(p-nitrophenyl)ρhosphate were reacted according to literature procedure (J. Med. Chem. 2000, 43, 1019-1028). The desired product was isolated in 65% yield (150 mg). ¹ H NMR (CDCIa, 300 MHz) δ 8.60 (s, IH), 8.00 (d, 2H, / = 7.8 Hz), 7.60 (t, IH, /= 7.2 Hz), 7.45 (app t, 2H, / = 7.7 Hz), 6.29 (d, IH, /= 1.5 Hz), 5.91-5.98 (m, IH), 5.67-5.71 (m, IH), 4.11-4.23 (m, 4H), 3.96-4.04

15 (m, 4H), 3.85-3.92 (m, IH), 2.55-2.72 (m, 2H), 1.29-1.37 (m, 6H). ³¹ P NMR (121.4 MHz, CDCl ₃ ) δ 19.8. LRMS (ESI) MH ⁺ C ₂₀ H ₂₇ N ₃ O ₉ P requires 484.1 Found 483.8.

Example 6

Scheme 8

20

O _n /=N O _n /=N

V-O HO u-n(^ ^O -e ' —V V^ ^NH ₂

HO ' Il

OBz OH

7.2 8.3

25

[5-(3-Carbamoyl-[l _/ 2,4]triazol-l-yl)-4-hydroxy-tetrahydro-furan-2- yloxymethyl]-phosphonic acid.

The l-[3-benzoyloxy-5-(diethoxy-phosphorylmethoxy)-tetrahydrofur an-2-yl]- 30 lH-[l,2,4]triazole-3-carboxylic acid methyl ester (60 mg. 0.124 mmol) was dissolved in MeCN (1.5 mL). Bromotrimethylsilane (0.164mL, 1.24 mmol) and 2,6-lutidine (0.043 mL, 0.372 mmol) were added and the mixture stirred at ambient temperature for 1O h. The solvents were removed in vacuo and the

262

5 product isolated by C-18 HPLC. To this was then added ammonia (7 N in MeOH, 4 mL) and the mixture stirred overnight. The title compound was isolated by C-18 HPLC. ¹ H NMR (300 MHz, D ₂ O) ppm 8.65 (s, IH), 5.95 (d, IH, / = 2.4 Hz), 5.49-5.52 (m, IH), 4.81-4.89 (m, IH), 3.47-3.70 (m, 2H), 2.27-2.32 (m, 2H). ³¹ P NMR (121.4 MHz, D2O) ppm 15.0. LRMS (ESI) M-H CeHi ₂ N ₄ O ₇ P requires 10 307.0 Found 307.0.

Example 7

Scheme 9

OH O O

NH ₃ HO- P H - NH, u HnO ^v '. W O 0 O OHH u HnO > f Ii

15 OH 'OH

8.3 9.4

Diphosphate of [5-(3-Carbamoyl-[l,2,4]triazol-l-yl)-4-hydroxy-tetrahydro-

20 furan-2-yloxymethyl]-phosphonic acid.

The [5-(3-carbamoyl-[l,2 _/ 4]triazol-l-yl)-4-hydroxy-tetrahydro-furan-2- yloxymethylj-phosphonic acid (20 mg, 0.058 mmol) was dissolved in anhydrous DMF (5 mL) and BusN (0.042 mL, 0.175 mmol) was added. The mixture was

25 concentrated in vacuo and then redissolved in anhydrous DMF (5 mL) containing N,N-carbonyldiimidazole (95.0 mg. 0.58 mmol). The mixture was stirred for 30 min and tributylammonium pyrophosphate (excess), predissolved in anhydrous DMF (1 mL) was added. After 2 h, the reaction was complete. Excess aqueous NH3 was added and the mixture concentrated in vacuo. The

30 desired product (9 mg) was isolated by ion exchange chromatography. ³¹ P NMR

263

5 (121.4 MHz, D ₂ O) ppm 7.10-7.30 (m), -6.26 (d, / = 20.9 Hz), -22.4 (app t, / = 21.4 Hz). LRMS (ESI) M-H C ₈ HuN ₄ Oi ₃ P ₃ requires 467.0 Found 466.9.

Example 8 Scheme 10

10

/y — \ O /=N

O ~ /=N HN \ / ^N J, _H

^Ho _H - _o ^°x!r ^N -N^ O ^NH2 — - -> ^{■ bH ' OH O}

8.3 9.5

15 Bis (Alanirte-n-butyl ester) phosphonamidate prodrug of [5-(3-carbamoyl- [l,2,4]triazol-l-yl)-4-hydroxy-tetrahydro-furan-2-yloxyineth yl]-phosphonic acid.

The [5-(3-carbainoyl-[l _/ 2 _/ 4]triazol-l-yl)-4-hydroxy-tetrahydro-furan-2- 20 yloxymethylj-phosphonic acid (21.7 mg, 0.057 mmol) was dissolved in anhydrous pyridine (3.2 mL) and the Alanine, n-butyl ester hydrochloride (72.4 mg, 0.40 mmol) was added. The mixture was concentrated in vacuo, redissolved in anhydrous pyridine and concentrated again. The resultant solids were suspended in anhydrous pyridine (2.2 mL) and stirred under Ar(g) at 60 ⁰ C. A 25 separate flask, PPha (104 mg, 0.40 mmol) and Aldrithiol (88.0 mg, 0.40 mmol) were combined in anhydrous pyridine (1 mL) and stirred for 20-30 min. The two mixtures were combined and stirred for 3-4 h at 60 ⁰ C under Ar(g). The mixture was concentrated and the product (10.6 mg) isolated by reverse-phase HPLC.

264

5 ³¹ P NMR (121.4 MHz, D ₂ O) ppm 21.6. LRMS (ESI) MH ⁺ C22H40N6O9P requires 563.3 Found 563.0.

Example 9 Scheme 30

O 0 NH ₂ . lf | ^H Il *t ^H 1) 2,4,6-triisopropyl I i HO P _OoEt , _n ¹ LA ₀ I X benzenesulfonyl chloride %A _O ^HO ^ ^PO3Et2 *y°-y» ^N ° TBDPSCI ^CL ^N O Et ₃ N, DMAP, MeCN _ «*-γ-°y* ^{N υ} _^

> — '., DMAP \ — / 2) NH ₄ OH -• — •. pyridinium

HO" 'OH TBSO° ''OTBS TBSO" 'OTBS p-toluene sulfonate

30.1 30.2 . 30.3

NH ₂ NH ₂ NH ₂ n ^?N fi (\ ^N

B _O ^O^°V^ ^NλO _{I N TBAFπ} - _{HF Et} α-?^^^ ^ _HO i^.o _v oλN ^λ c ^oE T ^t BS vd ¹ -L,'OTBS ^{^ OEt} : H;OM _ω O _H H issds; ^OH _H ^? _C fyX 'OH

30.4 30.5 30.6

V^CO ₂ Bu /^^^o^-^ (T ^ ^N

Aldrithiol-2, Ph ₃ P Py, 65 ⁰ C

10 30.7

l-((2R _/ 3R,4S)-3 _/ 4-bis(tert-butyldimethylsilyloxy)-5-methylene- tetrahydrofuran-2-yl)pyrimidine-2,4(lH,3H)-dione (30.2).

15 To a solution of compound 30.1 (540mg, 2.39mmol) in DMF (20 mL) was added imidazole (1.620, 23.9 mmol) and DMAP (290mg, 2.39mmol) and finally tert- butyldimethylsilyl chloride (2.15g, 14.3mmol). The mixture was heated to 6O ⁰ C for 4 h, water was added and the aqueous layer was extracted with ethyl acetate. The organic layer was dried over MgSO ₄ , filtered, and concentrated under

20 reduced pressure. The residue was subjected to a silica gel column

265

5 chromatography eluting with 40% EtOAc in Hexane to give compound 30.2 (700 mg, 67 % yield). ¹ H NMR (300 MHz, CDCb): δ 0.04 (s, 3H), 0.07 (s, 3H), 0.13 (s, 6H), 0.93 (s, 9H), 1.59 (s, 9H), 4.20 (dd, IH, / = 4.6, 1.0), 4.27 (d, IH, / = 4.3), 4.35 (d, IH, / = 4.3), 4.54 (s, IH), 5.80 (d, IH, / = 6.1), 6.08 (d, IH, /= 8.3), 7.18 (d, IH, / = 8.0), 8.24 (s, IH). LCMS [M-H]-C ₂ IH ₃₈ N ₂ OsSi ₂ requires 453.7. Found 453.3. 10

l-((2R,3R,4S)-3,4-bis(tert-butyldimethylsilyloxy)-5-methy lene- tetrahydrofuran-2-yl)-4-aminopyi±midin-2(lH)-one (30.3).

To a solution of compound 30.2 (940 mg, 2.07mmol) in acetonitrile (25 mL) was 15 added DMAP (505mg, 4.14mmol) and triethylamine (0.58mL, 4.14mmol) and 2,4,6 triisopropylbenzene sulfonyl chloride (1.25g, 4.14mmol). The mixture was stirred for 1 hour; concentrated ammonia hydroxide (20 mL) was added and stirred for 12 hours. The mixture was then concentrated under reduced pressure and the residue was partitioned between water and EtOAc. The organic layer 20 was dried over MgSO-t, filtered, and concentrated under reduced pressure. The residue was subjected to a silica gel column chromatography eluting with 5% methanol in CHaCbto give compound 30.3 (843 mg, 90% yield). ¹ H NMR (300 MHz, CDCb): δ 0.04 (s, 3H), 0.07 (s, 3H), 0.13 (s, 6H), 0.93 (s, 9H), 0.98 (s, 9H), 4.22 (s, IH), 4.27 (d, IH, J = 2.1), 4.35 (d, IH, / = 4.3), 4.58 (s, IH), 5.78 (d, IH, / = 25 8.1), 5.95 (d, IH, / = 2.3), 7.22 (d, IH, / = 8.1).

diethyl((2R,3S,4R,5R)-5-(4-amino-2-oxopyrimidin-l(2H)-yl) -3,4-bis(tert- butyldiinethylsilyloxy)-2-rnetiιyl-tetraLhydτofuraiι-2-yl oxy)inethylphosphonate (30.4). 30

To a solution of compound 30.3 (840mg, 1.85mmol) in dichloroethane (2OmL) was added diethyl (hydroxymethyl) phosphonate (0.41 mL, 2.87 mmol) and pyridinium p-toluene sulfonate (0.23g, 0.92mmol). The mixture was heated to

266

5 60 ⁰ C for 1 h and then concentrated under reduced pressure. The residue was subjected to a silica gel column chromatography eluting with 10% MeOH in CH2CI2 to give compound 30.4 (289 mg, 24% yield). ¹ H NMR (300 MHz, CDCl ₃ ): δ®0.04 (s, 3H), 0.08 (s, 3H), 0.10 (s, 6H), 0.89 (s, 9H), 0.92 (s, 9H), 1.38 (m, 6H), 1.49 (s, 3H), 3.93 (m, IH), 4.14 (m, 2H), 4.20 (m, IH), 4.38 (m, IH), 5.83 (d, IH, / = 7.7), 10 6.08 (d, IH, / = 4.0), 7.99 (d, IH, / = 7.3). LRMS (ESI) MH ⁺ C ₂₆ H ₅₂ N ₃ OsPSi ₂ requires 622.9. Found 622.1.

diethyl((2R,3S,4R,5R)-5-(4-amino-2-oxoρyrimidin-l(2H)-yl )-3,4-dihydroxy-2- methyl-tetrahydrofuran-2-yloxy)methylphosphonate (30.5).

15

To a solution of compound 30.4 (288 mg, 0.46mmol) in THF (20 mL) was added a IN solution of tetrabutylammonium fluoride in THF (2.7mL, 2.8mmol). The mixture was stirred at r.t. for 1 h and concentrated down under reduced pressure. The residue was subjected to a silica gel column chromatography

20 eluting with 10% MeOH in CH2CI2 to give compound 30.5 (114mg, 63% yield). ¹ H NMR (300 MHz, CD ₃ OD): δ 1.38 (t, 6H, / = 7.0), 1.40 (s, 3H), 3.86 (m, 2H), 4.02 (d, IH, / = 4.9), 4.11 (m, 4H), 4.56 (dd, IH, / = 7.0, 6.7), 5.98 (d, IH, / = 7.4), 6.11 (d, IH, / = 7.0), 7.63 (d, IH, / = 7.6). LRMS (ESI) MH ⁺ Ci ₄ H ₂₄ NsO ₈ P requires 394.4. Found 394.1.

25

((2R,3S,4R,5R)-5-(4-amino-2-oxopyriirιidin-l(2H)-yl)-3,4 -dihydroxy-2-inethyl- tetrahydrofuran-2-yloxy)methylphosphonic acid (30.6).

To a solution of compound 30.5 (112 mg, 0.29 mmol) in acetonitrile (10 mL) was 30 added 2, 6-lutidine (0.20 mL, 1.7 mmol) and then trimethylsilyl iodide (0.41 mL, 2.8 mmol). The mixture was stirred at room temperature for 1 h. After the reaction was done, 0.30 mL of 2, 6-lutidine was added to the mixture and the mixture was concentrated under reduced pressure. The residue was then put

267

5 into solution with water (3 mL) and treated with sodium hydroxide IN to pH=10. The solution was then acidify to pH=3 using acetic acid and subjected to a reverse phase (YMC-Pack ODS-A) to give compound 30.6 (69 mg, 72% yield). ¹ H NMR (300 MHz, DaO): δ 1.39 (s, 3H), 3.54 (m, 2H), 3.99 (d, IH, / = 4.6), 4.63 (m, IH), 6.09 (d, IH, 7 = 7.0), 6.14 (d, IH, / = 7.0), 8.0 (d, IH, / = 7.6). LRMS (ESI) MH ⁺ 10 CioHi6N3θβP requires 338.2. Found 337.8.

Bis-amidate prodrug 30.7.

To a solution of compound 30.6 (31.7 mg, 0.094mmol) in pyridine (2 mL) was added the bis-alabutylamine (85.0mg, 0.47mmol), triphenylphosphine (148 mg,

15 0.56mmol), triethylamine (80 uL, 0.47mmol) and Aldrichthiol-2 (104 mg, 0.47 mmol). The mixture was heated to 6O ⁰ C for 3 h and then concentrated down under reduced pressure. The residue was diluted in ethyl acetate and washed with a IN solution of NaHzPCk and then with brine. The organic layer was dried over Na-SO-J, filtered, and concentrated under reduced pressure. The residue

20 was subjected to a silica gel column chromatography eluting with 10% MeOH in. CH2CI2 to give compound 30.7 (llmg, 20% yield).

25

30

35

268

Example 10

Scheme 31

NH ₂ NH ₂ NH ₂

( T J < 1 J HO^PO ₃ Et ₂ p < A J

N-/ DMAP >— / pyridinium ^0Et ^ ) — (

HO ^V 'OH TBSd ^* 'OTBS p-to!uene sulfonate TBSO OTBS

31.1 31.2 ³¹ - ³

NH ₂ NH ₂

N^λ _>N N-^- ¹ W _N

^1NTBAF/TH,F _ao W OEt *'" \ y / ^{N N';} 2 ™,6- ^s lu ^ι tidiπ ^, e HorJ OUH *v° \ S j^ ^N ^ wf ^'' '0H HO° ^' 'OH

31.4 31.5

9-((2R,3R,4S)-3,4-bis(tert-butyldimethylsilyloxy)-5-methy lene- 10 tetrahydrofuran-2-yl)-9H-ptirin-6-ainine (31.2).

To a solution of compound 31.1 (276mg, 2.39mmol) in DMF (10 mL) was added imidazole (604 mg, 8.9 mmol) and DMAP (135mg, 1.Immol) and finally tert- butyldimethylsilyl chloride (665 mg, 4.4mmol). The mixture was heated to 60 ⁰ C

15 for 4 h, water was added and the aqueous layer was extracted with ethyl acetate. The organic layer was dried over MgSCX filtered, and concentrated under reduced pressure. The residue was subjected to a silica gel column chromatography eluting with 50% EtOAc in Hexane to give compound 31.2 . (280mg, 63 % yield). ¹ H NMR (300 MHz, CDCl ₃ ): δ -0.30 (s, 3H), -0.06 (s, 3H), 0.16

20 (s, 6H), 0.77 (s, 9H), 0.96 (s, 9H), 4.30 (s, IH), 4.54 (s, IH), 4.60 (d, IH, / = 4.2), 5.10 (t, IH), 5.52 (s, 2H), 6.12 (d, IH, J = 5.8), 7.89 (s, IH), 8.39 (s, IH). LRMS (ESI) MH ⁺ CazHagNsOaSia requires 478.8. Found 477.9.

269

5 diethyl ((2R,3S,4R,5R)-5-(6-amino-9H-purin-9-yl)-3,4-bis(tert- bufyldiinethyIsilyloxy)-2-inethyl-tetrahydrofuran-2-yloxy)in ethylphosphonate (31.3).

To a solution of compound 31.2 (280 mg, 0.59 mmol) in dichloroethane (5 mL) 10 was added diethyl (hydroxymethyl) phosphonate (0.22 mL, 1.47 mmol) and pyridinium p-toluene sulfonate (56 mg, 0.29 mmol). The mixture was heated to 6O ⁰ C for 1 h and then concentrated under reduced pressure. The residue was subjected to a silica gel column chromatography eluting with 10% MeOH in CH2CI2 to give compound 31.3 (102 mg, 16% yield). ¹ H NMR (300 MHz, CDCl ₃ ): 15 δ -0.45 (s, 3H), -0.08 (s, 3H), 0.05 (s, 3H), 0.71 (s, 9H), 0.98 (s, 9H), 1.35 (m, 6H), 1.61 (s, 3H), 3.78 (t, IH), 3.95 (t, IH), 4.04 (s, IH), 4.21 (m, 4H), 4.96 (m, IH), 6.24 (d, IH, / = 6.7), 8.10 (s, IH), 8.70 (s, IH). LRMS (ESI) MH ⁺ C ₂₇ H ₅₂ NsO ₇ PSi ₂ requires 646.9. Found 645.9.

diethyl ((2R,3S,4R,5R)-5-(6-ajmino-9H-purin-9-yl)-3,4-dihydroxy-2-me thyl- 20 tetrahydrofuran-2-yloxy)methylphosphonate (31.4).

To a solution of compound 31.3 (102 mg, 0.16mmol) in THF (5 mL) was added a IN solution of tetrabutylammonium fluoride in THF (0.15 mL, 0.5 mmol). The mixture was stirred at r.t. for 1 h and concentrated down under reduced 25 pressure. The residue was subjected to a silica gel column chromatography eluting with 10% MeOH in CH2CI2 to give compound 31.4 (38 mg, 59% yield). ¹ H NMR (300 MHz, CD ₃ OD): δ 1.28 (m, 6H), 1.52 (s, 3H), 3.62 (m, IH), 3.80 (m, 2H), 4.02 (m, 4H), 4.11 (m, IH), 6.15 (d, IH, / = 7.1), 8.1 (s, IH), 8.75 (s, IH). LRMS (ESI) MH ⁺ Ci ₅ H ₂₄ NsO ₇ P requires 418.4. Found 417.9.

30 ((2R,3S,4R,5R)-5-(6-amino-9H-purin-9-yI)-3,4-dihydroxy-2-met hyl~ tetrahydrofuran-2-yloxy)methylphosphonic acid (31.5).

270

5 To a solution of compound 31.4 (20 mg, 0.048 mmol) in acetonitrile (2 mL) was added 2, 6-lutidine (0.034 mL, 0.29 mmol) and then trimethylsilyl iodide (0.068 mL, 0.48 mmol). The mixture was stirred at room temperature for 1 h. After the reaction was done, 0.08 mL of 2, 6-lutidine was added to the mixture and the mixture was concentrated under reduced pressure. The residue was then put

10 into solution with water (2 mL) and treated with sodium hydroxide IN to pH=10. The solution was then acidify to pH=3 using acetic acid and subjected to a reverse phase (YMC-Pack ODS-A) to give compound 31.5 (69 mg, 72% yield). ¹ H NMR (300 MHz, D2O): δ 1.42 (s, 3H), 3.33 (t, IH, J = 10.7), 3.51 (t, IH, / = 11.9), 4.12 (d, IH, /= 4.6), 5.18 (m, IH), 6.10 (d, IH, / = 7.1), 8.24 (s, IH), 8.55 (s, IH).

15 LRMS (ESI) MH ⁺ CπHieNsO/P requires 362.3. Found 361.9.

Example 11

Scheme 32

^{^} NH 11 r

_C y^o HO. -PO ₃ Et ₂ O

EtO r- °P-

OEt λ _^ J EtO ,'~pT'— ^ o-t-V

HO ¹ 'OH pyridinium H(f ^' 'OH ^OEt HO* ^{" '} ''OH

32.1 p-toluene sulfonate 32.2 32.3

O

V^CO ₂ Bu NH ₂ HCI

AIdrithiol-2, Ph ₃ P Py ₁ 65 ⁰ C

20

271

5 diethyl ((2R _/ 3S _r 4R _λ 5R)-5-(2,4-dioxo-3 _/ 4-dihydropyrimidiα-l(2H)-yl)-3,4- dihydroxy-2-methyl-tetrahydrofuran-2-yloxy)methylphosphonate (32.2); diethyl ((2S,3S _/ 4R _/ 5R ⁾ -5- ⁽ 2 _/ 4-dioxo-3,4-dihydropyrimidm-l(2H)-yl ⁾ -3 _/ 4- dihydroxy-2-methyl-tetrahydrofuran-2-yloxy)inethylphosphonat e (32.3).

10 To a solution of compound 32.1 (0.67 g, 2.8 mmol) in dimethyl formamide (15 mL) was added diethyl (hydroxymethyl) phosphonate (0.65 mL, 4.4 mmol) and pyridinium p-toluene sulfonate (0.37 g, 1.5 mmol). The mixture was heated to 80 ⁰ C for 1 h and then concentrated under reduced pressure. The residue was subjected to a silica gel column chromatography eluting with 10% MeOH in

15 CH2CI2 and then 10% MeOH in EtOAc to give two diastereoisomers, compound 32.2 (80 mg, 7% yield) and compound 32.3 (100 mg, 8.5% yield). Data for compound 32.2: ¹ H NMR (300 MHz, CD ₃ OD): 1.37 (t, 6H), 1.52 (s, 3H), 3.95 (m, IH), 4.02 (m, 2H), 4.10 (m, 2H), 4.25 (m, 4H), 5.71 (d, IH, / = 8.0), 5.85 (d, IH, / = 2.7), 7.56 (d, IH, / = 8.0). LRMS [M-H]- C14H23N2O9P requires 393.3. Found 393.2.

20

((2R,3S,4R,5R)-5-(2,4-dioxo-3,4-dihydroρyrimidin-l(2H)-y l)-3,4-dihydroxy-2- m.ethyl-tetrahydrofuran-2-yloxy)methylphosphonic acid (32.4).

Compound 32.4 (61 mg, 75% yield) was synthesized from compound 32.2 (95 mg, 25 0.24 mmol) using the procedure described for the preparation of compound 31.5. ¹ H NMR (300 MHz, D ₂ O) δ 1.38 (s, 3H), 3.50 (m, 2H), 3.98 (d, IH, / = 4.6), 5.84 (d, IH, / = 7.6), 6.08 (d, IH, / = 7.3), 7.81 (d, IH, / = 8.2). LRMS [M-H]- C10H15N2O9P requires 337.2. Found 337.2.

30 Bisamidate prodrug 32.5.

Compound 32.5 (6.2 mg, 18% yield) was synthesized from compound 32.4 (19.4 mg, 0.057 mmol) using the procedure described for the preparation of compound 30.7. ¹ H NMR (300 MHz, CDsOD): δ 0.98 (t, 6H, / = 7.3), 1.34-1.46 (m,

272

5 8H), 1.44 (s, 3H), 1.66 (m, 4H), 3.67 (m, 2H), 3.91 (d, IH, / = 4.5), 4.18 (m, 4H), 4.56 (m, IH), 5.69 (d, IH, / = 8.3), 6.07 (d, IH, / = 7.4), 7.93 (d, IH, / = 8.2).

HCV Replicon and Cytotoxicity Assays

10 The compounds of the present invention can be evaluated using known

HCV replicon assays and cytotoxicity assays, for example those described in Stuyver et al _v Antimicrobial Agents and Chemotherapy 2003, 47(1), pp. 244-254, and Lohman et al., Science 1999, vol.285, pp. 110-113, each of which is herein incorporated by reference in its entirety. Exempliery experimental protocols are

15 shown below.

HCV ICso Determination

Assay Protocol: NS5b polymerase assay (40 μL) was assembled by adding 28 μL polymerase mixture (final concentration: 50 mM Tris-HCl at pH 7.5, 10 niM KCL, 5 mM MgCl ₂ , 1 mM DTT, 10 mM EDTA, 4 ng/μL of RNA template,

20 and 75 nM HCV ®21 NS5b polymerase) to assay plates followed by 4 μL of compound dilution. The polymerase and compound were pre-incubated at 35 ⁰ C for 10 minute before the addition of 8 μL of nucleotide substrate mixture (33P-@-labeled competing nucleotide at KM and 0.5 mM of the remaining three nucleotides). The assay plates were covered and incubated at 35 ⁰ C for 90 min.

25 Reactions were then filtered through 96-well DEAE-81 filter plates via vacuum. The filter plates were then washed under vacuum with multiple volumes of 0.125 M NaHPCU, water, and ethanol to remove unincorporated label. Plates were then counted on TopCount to assess the level of product synthesis over background controls. The IC50 value was determined using Prism fitting

30 program.

Table 2E shows representative examples of the activity of the compounds of the invention at a particular concentration when tested in this assay.

273

Table 2E. Representative activity of compounds in the polymerase assay.

10 HCV ECso Determination

Replicon cells were seeded in 96-well plates at a density of 8 x 10 ³ cells per well in 100 μL of culture medium, excluding Geneticin. Compound was serially diluted in 100% DMSO and then added to the cells at a 1:200 dilution, achieving a final concentration of 0.5% DMSO and a total volume of 200 μL. Plates were

15 incubated at 37°C for 3 days, after which culture medium was removed and cells were lysed in lysis buffer provided by Promega's luciferase assay system. Following the manufacturer's instruction, 100 μL of luciferase substrate was added to the lysed cells and luciferase activity was measured in a TopCount luminometer.

20 Typically, compounds of the invention that were tested were found to have an ECso of less than about 1000 μM (Huh7). Some compounds demonstrated an ECso of less than about 250 μM (Huh7).

Table 3E shows representative examples of the activity of the compounds of the invention at a particular concentration when tested in this assay.

25

274

5 Table 3E. Representative activity of compounds in the replicon assay.

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

10 Cytotoxicity Cell Culture Assay (Determination of CC50):

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

1. Maintain MT-2 cells in RPMI-1640 medium supplemented with 5% fetal 15 bovine serum and antibiotics.

2. Distribute the cells into a 96-well plate (20,000 cell in 100 μl media per well) and add various concentrations of the tested compound in triplicate (100 μl/well). Include untreated control.

3. Incubate the cells for 5 days at 37 °C.

20 4. Prepare XTT solution (6 ml per assay plate) in dark at a concentration of

2mg/ml in a phosphate-buffered saline pH 7.4. Heat the solution in a water- bath at 55°C for 5 min. Add 50 μl of N-methylphenazonium methasulfate (5 μg/ml) per 6 ml of XTT solution.

275

5 5. Remove 100 μl media from each well on the assay plate and add 100 μl of the XTT substrate solution per well. Incubate at 37 ⁰ C for 45 to 60 min in a CO2 incubator. 6. Add 20 μl of 2% Triton X-100 per well to stop the metabolic conversion of

XTT. 10 7. Read the absorbance at 450 ran with subtracting off the background at 650 run.

8. Plot the percentage absorbance relative to untreated control and estimate the CC50 value as drug concentration resulting in a 50% inhibition of the cell growth. Consider the absorbance being directly proportional to the cell 15 growth.

276