Note: Descriptions are shown in the official language in which they were submitted.
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Piperidine Compounds
Background of the Invention
Field of the Invention
Neuraminidase (also known as sialidase, acylneuraminyl hydrolase,
and EC 3.2.1.18) is an enzyme common among animals and a number of
microorganisms. It is a glycohydrolase that cleaves terminal alpha-
ketosidically linked sialic acids from glycoproteins, glycolipids and
oligiosaccharides. Many of the microorganisms containing neuraminidase are
pathogenic to man and other animals including fowl, horses, swine and seals.
Organisms having N-acetylneuraminidases include bacteria such as Vibrio
cholerae, C. perfringens and Streptococcus sp. and viruses such as influenza
virus, and parainfluenza virus.
Influenza neuraminidase has been implicated in the pathogenicity of
influenza viruses. It is thought to help the elution of newly synthesized
virons from infected cells and assist in the movement of the virus (through
its hydrolase activity) through the mucus of the respiratory tract.
Brief Description of Related Art
von Itzstein, L. M. et al., "Nature", 363(6428):418-423 (1993), discloses the
rational design of sialidase-based inhibitors of influenza virus replication.
Colman, P. M. et al. International Patent Publication No. WO 92/06691
(Int. App. No. PCT/AU90/00501, publication date April 30, 1992), von Itzstein,
L. M. et al., European Patent Publication No. 0 539 204 Al (EP App. No.
92309684.6, publication date April 28, 1993), and von Itzstein, L. M. et al.
International Publication No. WO 91/16320 (Int. App. No. PCT/AU91/00161,
publication date October 31, 1991) disclose compounds that bind
neuraminidase and are asserted to exhibited antiviral activity in vivo.
Umezawa, H. et al., "J. Antibiotics" 27:963-969 (1974), discloses the
isolation of Siastatin B. Nishimura, Y. et al., "J. Am. Chem. Soc." 110:7249-
7250
(1988), and "Bull. Chem. Soc. Jpn." 65:978-986 (1992), disclose the total
synthesis
of Siastatin B. Nishimura, Y. et al. "J. Antibiotics" 45(10):1662-1668 (1992);
46(2):
300-309 (1993); 46(12):1883-1889 (1993); 47(1):101-107 (1994); and "Nat. Prod.
Lett." 1(1): 39-44 (1992); as well as Japanese Patent Applications 92-287381
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WO 98/17647 PCTIUS97/18193
(October 26, 1992); 90-201437 (July 31, 1990); 88-125020 (May 24, 1988) and
50046895 (April 25, 1975) disclose synthetic transformations of Siastatin B
including certain dehydrosiastatin B analogs. Zbiral, E. et al., "Liebigs Ann.
Chem." 129-134 (1991), and von Itzstein, L. M. et al., "Carbohydrate Res."
244:181-185 (1993), disclose synthetic transformation of the hydroxy group at
C4
of sialic acid to an amino group.
Objects of the Invention
Selected embodiments of the invention satisfy one or more of the
following objects:
A principal object of the invention is inhibition of bacteria and viruses,
in particular influenza viruses. In particular, an object is inhibition of
glycolytic enzymes such as neuraminidase, in particular the selective
inhibition of viral or bacterial neuraminidases.
An additional object of the invention is to provide neuraminidase
inhibitors that have a retarded rate of urinary excretion, that enter into
nasal
or pulmonary secretions from the systemic circulation, that have sufficient
oral bioavailability to be therapeutically effective, that possess elevated
potency, that exhibit clinically acceptable toxicity profiles and have other
desirable pharmacologic properties.
Another object is to provide improved and less costly methods for
synthesis of neuraminidase inhibitors.
A still further object is to provide improved methods for
administration of known and novel neuraminidase inhibitors.
An additional object is to provide compositions useful in preparing
polymers, surfactants or immunogens and for use in other industrial
processes and articles.
These and other objects will be readily apparent to the ordinary artisan
from consideration of the invention as a whole.
Summary of the Invention
Compounds, or compositions having formula (IX) are provided herein:
J2 J2
W6N-, N E1
T1 J1a
J2
G1 2a
-2-
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wherein
Ei is -(CR1R1)m1W1;
G1 is N3, -CN, -OH, -OR6a, -N02, or -(CR1R1)m1W2;
T1 is -NR1W3, or a heterocycle;
Jla are independently Ri, Br, Cl, F, I, CN, N02 or N3;
J2 and J2a are independently H or Rl;
Rl is independently H or alkyl of 1 to 12 carbon atoms;
R2 is independently R3 or R4 wherein each R4 is independently
substituted with 0 to 3 R3 groups;
R3 is independently F, Cl, Br, I, -CN, N3, -NO2, -ORba, -OR1, -N(Rl)2,
-N(R1)(R6b). -N(R6b)2. -SR1, -SR6a, -S(O)R1, -S(0)2R1, -S(O)OR1, -S(O)OR6a.
-S(O)20R1, -S(0)20R6a, -C(O)OR1, -C(O)R6c, -C(O)OR6a, -OC(O)Ri,
-N(Rl)(C(O)Rl), -N(R6b)(C(O)Rl), -N(Rl)(C(O)ORl), -N(R6b)(C(O)ORl),
-C(O)N(Rl)2, -C(O)N(R6b)(R1), -C(O)N(R6b)2, -C(NR1)(N(R1)2).
-C(N(R6b))(N(Rl)2), -C(N(Rl))(N(R1)(R6b)), -C(N(R6b))(N(Rl)(R6b)),
-C(N(R1))(N(R6b)2), -C(N(R6b))(N(R6b)2), -N(R1)C(N(R1))(N(Rl)2),
-N(Rl)C(N(Rl))(N(Rl)(R6b)), -N(R1)C(N(R6b))(N(Rl)2),
-N(R6b)C(N(R1))(N(R1)2), -N(R6b)C(N(R6b))(N(Rl)2),
-N(R6b)C(N(R1))(N(R1)(R6b)), -N(Rl)C(N(R6b))(N(R1)(R6b)).
-N(Rl)C(N(R1))(N(R6b)2), -N(R6b)C(N(R6b))(N(R1)(R6b)),
-N(R6b)C(N(R1))(N(R6b)2), -N(R1)C(N(R6b))(N(R6b)2),
-N(R6b)C(N(R6b))(N(R6b)2), =0, =S, =N(Rl) or =N(R6b);
R4 is independently alkyl of 1 to 12 carbon atoms, alkenyl of 2 to 12
carbon atoms, or alkynyl of 2 to 12 carbon atoms;
R5 is independently R4 wherein each R4 is substituted with 0 to 3 R3
groups;
R5a is independently alkylene of 1 to 12 carbon atoms, alkenylene of 2 to
12 carbon atoms, or alkynylene of 2-12 carbon atoms any one of which
alkylene, alkenylene or alkynylene is substituted with 0-3 R3 groups;
R6a is independently H or an ether- or ester-forming group;
R6b is independently H, a protecting group for amino or the residue of a
carboxyl-containing compound;
R6c is independently H or the residue of an amino-containing
compound;
Wi is a group comprising an acidic hydrogen, a protected acidic group,
or an R6c amide of the group comprising an acidic hydrogen;
W2 is a group comprising a basic heteroatom or a protected basic
- 3-
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heteroatom, or an R6b amide of the basic heteroatom;
W3 is W4 or W5;
W4 is R5 or -C(O)R5, -C(O)W5, -S02R5, or -S02W5;
W5 is carbocycle or heterocycle wherein W5 is independently
substituted with 0 to 3 R2 groups;
W6 is -R5, -W5, -R5aW5, -C(O)OR6a, -C(O)R6c, -C(O)N(R6b)2,
-C(NR6b)(N(R6b)2), -C(NR6b)(N(H)(R6b)), -C(N(H)(N(R6b)2), -C(S)N(R6b)2, or
-C(O)R2; and
each m1 is independently an integer from 0 to 2;
provided, however, that compounds are excluded wherein Jla is H, each J2 is
H, J2a is H and Tl is -N(H)(Ac) and:
E1 is -CO2H or -CO2CH3,
G1 is -OBoc, and
W6 is Boc;
E1 is -CO2H or -CO2CH3,
G1 is -OH, and
W6 is H;
E1 is -CO2H, -CO2CH3 or -CO2Bn
G1 is -OH, and
W6 is Boc;
E1 is -CONH2,
G1 is -OH, and
W{, is Boc or H;
El is -CO2H or -CO2CH3,
Gl is OH, and
W6 is Bn; or
Et is -CO2H or -CO2CH3,
G1 is -OH, and
W6 is -CH2CH(OH)CH2(OH);
wherein Bn is benzyl and Boc is -CO2C(CH3)3;
and the salts, solvates, resolved enantiomers and purified diastereomers
thereof.
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In another embodiment, compounds, or compositions having formula
(X) are provided herein:
Z2
Z1-N J1
1 E1
(X)
JT, N Jla
J2 Z1 1
wherein
one Zl is W6 and the other Zl is G1;
Z2 is H or W6;
El is -(CR1R1)m1Wl;
G1 is -OH, -OR6a, or -(CR1R1)m1W2;
T1 is -NR1W3 or a heterocycle;
Ji and J1a are independently Rl, Br, Cl, F, I, CN, N02 or N3;
J2 is H or Rl;
Rl is independently H or alkyl of 1 to 12 carbon atoms;
R2 is independently R3 or R4 wherein each R4 is independently
substituted with 0 to 3 R3 groups;
R3 is independently F, Cl, Br, I, -CN, N3, -N02, -OR6a, -OR1, -N(Rl)2,
-N(Rl)(R6b), -N(R6b)2, -SR1, -SR6a, -S(O)Rl, -S(0)2R1, -S(O)OR1, -S(O)OR6a,
-S(O)20R1, -S(0)2OR6a, -C(O)OR1, -C(O)R6c, -C(O)OR6a, -OC(O)Rl,
-N(R1)(C(O)R1), -N(R6b)(C(O)Rl), -N(Rl)(C(O)OR1), -N(R6b)(C(O)OR1),
-C(O)N(R1)2, -C(O)N(R6b)(Rl), -C(O)N(R6b)2, -C(NRl)(N(R1)2),
-C(N(R6b))(N(R1)2), -C(N(R1))(N(Rl)(R6b)), -C(N(R6b))(N(R1)(R6b)),
-C(N(R1))(N(R6b)2), -C(N(R6b))(N(R6b)2), -N(R1)C(N(R1))(N(R1)2),
-N(Rl)C(N(Rl))(N(RZ)(R6b)), -N(R1)C(N(R6b))(N(R1)2),
-N(R6b)C(N(R1))(N(R1)2), -N(R6b)C(N(R6b))(N(R1)2),
-N(R6b)C(N(R1))(N(Rl)(R6b)), -N(Rl)C(N(R6b))(N(RZ)(R6b)).
-N(Rl)C(N(Rl))(N(R6b)2), -N(R6b)C(N(R6b))(N(R2)(R6b)).
-N(R6b)C(N(Rl))(N(R6b)2), -N(R1)C(N(R6b))(N(R6b)2),
-N(R6b)C(N(R6b))(N(R6b)2), =0, =S, =N(RI) or =N(R6b);
R4 is independently alkyl of 1 to 12 carbon atoms, alkenyl of 2 to 12
carbon atoms, or alkynyl of 2 to 12 carbon atoms;
R5 is independently R4 wherein each R4 is substituted with 0 to 3 R3
groups;
R5a is independently alkylene of 1 to 12 carbon atoms, alkenylene of 2 to
12 carbon atoms, or alkynylene of 2-12 carbon atoms any one of which
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alkylene, alkenylene or alkynylene is substituted with 0-3 R3 groups;
R6a is independently H or an ether- or ester-forming group;
R6h is independently H, a protecting group for amino or the residue of a
carboxyl-containing compound;
R6c is independently H or the residue of an amino-containing
compound;
Wi is a group comprising an acidic hydrogen, a protected acidic group,
or an R6c amide of the group comprising an acidic hydrogen;
W2 is H or a group comprising a basic heteroatom or a protected basic
heteroatom, or an R6b amide of the basic heteroatom;
W3 is W4 or W5;
W4 is R5 or -C(O)R5, -C(O)W5, -S02R5, or -S02W5;
W5 is carboc cle or heteroc cle wherein W5 y y is independently
substituted with 0 to 3 R2 groups;
W6 is -R5, -W5, -R5aW5, -C(O)OR6a, -C(O)R6c, -C(O)N(R6b)2,
-C(NR6b)(N(R6b)2), -C(NR6b)(N(H)(R6b)), -C(N(H)(N(R6b)2), -C(S)N(R6b)2, or
-C(O)R2;
each ml is independently an. integer from 0 to 2;
and the salts, solvates, resolved enantiomers and purified diastereomers
thereof.
The present invention as claimed provides a compound of formula
(XXVIII) or (XXVIIa):
ws Q J1 Ws 0 J2 J2a
J2 ~ Ei J2 El
(XXVIII) I (XXVIIla)
Ty J2 N J2 2a T~ J N Ji
G1 2 G,
wherein:
E1 is -(CR1 R1)m1 W1 ;
6
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G1 is W2 or -NHR1, -C(NH)(NH2), -NR1-C(NR1)(NR1 R3), -NH-C(NH)(NHR3),
-NH-C(NH)(NHR1), -NH-C(NH)NH2, -CH(CH2NHR1)(CH2OH),
-CH(CH2NHR1)(CH2NHR1), -CH(NHR1)-(CR1 R1)m2-CH(NHR1)R1,
-CH(OH}{CR1 R1)m2-CH(NHR1)R1, or -CH(NHR1 }(CR1 R1)m2-CH(OH)R1,
-(CR1 R1)m2-S-C(NH)NH2, -N=C(NHR1)(R3), -N=C(SR1)N(R1)2,
-N(R1)C(NH)N(R1)C=N, or -N=C(NHR1)(R1);
T1 is -NR1 W3, or a monocycle having 3 to 7 ring members containing 2
to 6 carbon atoms and 1 to 3 heteroatoms selected from N, 0, P and S,
or a bicycle having 7 to 10 ring members containing 4 to 9 carbon atoms
and 1 to 3 heteroatoms selected from N, 0, P and S;
J1 is R1, Br, Cl, F, I, CN, NO2 or N3;
J2 is H or R1;
J2a is H or R1;
R1 is independently H or alkyl of 1 to 12 carbon atoms;
R2 is independently R3 or R4 wherein each R4 is independently
substituted with 0 to 3 R3 groups;
R3 is independently F, Cl, Br, I, -CN, N3, -N02, -OR6a, -OR1, -N(R1)2,
-N(R1)(R6b), -N(R6b)2, -SR1, -SR6a, -S(O)R1, -S(O)2R1, -S(O)OR1, -S(O)OR6a,
-S(O)20R1, -S(O)2OR6a, -C(O)OR1, -C(O)R6c, -C(O)OR6a, -OC(O)R1,
-N(R1)(C(O)R1), -N(R6b)(C(O)R1), -N(R1)(C(O)OR1), -N(R6b)(C(O)OR1),
-C(O)N(R1)2, -C(O)N(R6b)(R1), -C(O)N(R6b)2, -C(NR1)(N(R1)2),
-C(N(R6b))(N(R1)2), -C(N(R1))(N(R1)(R6b)), -C(N(R6b))(N(R1)(R6b)).
-C(N(R1))(N(R6b)2), -C(N(R6b))(N(R6b)2), -N(R1)C(N(R1))(N(R1)2),
-N(R1)C(N(R1))(N(R1)(R6b)), -N(R1)C(N(R6b))(N(R1)2),
-N(R6b)C(N(R1))(N(R1)2), -N(R6b)C(N(R6b))(N(R1)2),
-N(R6b)C(N(R1))(N(R1)(R6b)2), -N(R1)C(N(R6b))(N(R1)(R6b)),
-N(R1)C(N(R1))(N(R6b)2), -N(R6b)C(N(R6b))(N(R1)(R6b)),
6a
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N(R6b)C(N(R1))(N(R6b)2), -N(R1)C(N(R6b))((N(R6b)2),
-N(R6b)C(N(R6b))(N(R6b)2), =0, =S, =N(R1), =N(R6b) or W5;
R3a is independently -CN, N3, -NO, -NO2, -OR6a, -OR1, -N(R1)2,
-N(R1)(R6b), -N(R6b)2, -SR1, -SR6a, -S(O)R1, -S(O)2R1, -S(O)OR1,
-S(O)OR6a, -S(O)20R1, -S(O)2OR6a, -C(O)OR1, -C(O)R6c, -C(O)OR6a,
-OC(O)R1, -N(R1)(C(O)R1), -N(R6b)(C(O)R1), -N(R1)(C(O)OR1),
-N(R6b)(C(O)OR1), -C(O)N(R1)2, -C(O)N(R6b)(R1), -C(O)N(R6b)2,
-C(NR1)(N(R1)2), -C(N(R6b))(N(R1)2), -C(N(R1))(N(R1)(R6b)),
-C(N(R6b))(N(R1)(R6b)), -C(N(R1))(N(R6b)2), -C(N(R6b))(N(R6b)2),
-N(R1)C(N(R1))(N(R1)2), -N(R1)C(N(R1))(N(R1)(R6b)), -N(R1)C(N(R6b))(N(R1)2),
-N(R6b)C(N(R1))(N(R1)2), -N(R6b)C(N(R6b)XN(R1)2), -N(R6b)C(N(R1)XN(R1 XR6b)),
-N(R1)C(N(R6b)XN(R1 XR6b)), -N(R1)C(N(R1)XN(R6b)2), -f'1(R6b)C((N(R6b))N(R1
XR6b)),
-N(R6b)C(N(R1)XN(R6b)2), -N(R1)C(N(R6b))(N(R6b)2) or
-N(R6b)C(N(R6b))N(R6b)2),
R4 is independently alkyl of 1 to 12 carbon atoms, alkenyl of 2 to 12
carbon atoms, or alkynyl of 2 to 12 carbon atoms;
R5 is independently R4 wherein each R4 is substituted with 0 to 3 R3
groups;
R5a is independently alkylene of 1 to 12 carbon atoms, alkenylene of 2 to
12 carbon atoms, or alkynylene of 2-12 carbon atoms any one of which alkylene,
alkenylene or alkynylene is substituted with 0-3 R3 groups;
R6a is independently H, an ether-group or an ester group;
R6b is independently H, a protecting group for amino or the residue of a
carboxyl-containing compound;
R6c is independently H or the residue of an amino-containing compound;
W1 is a group comprising an acidic hydrogen;
W2 is a group comprising a N or S atom;
6b
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W3 is W4 or W5;
W4 is R5 or -C(O)R5, -C(O)W5, -S02R5, or -S02W5;
W5 is carbocycle selected from a monocycle containing 3 to 7 carbon
atoms and a bicycle containing 7 to 12 carbon atoms; or a heterocycle selected
from a monocycle having 3 to 7 ring members containing 2 to 6 carbon atoms
and 1 to 3 heteroatoms selected from N, 0, P and S, and a bicycle having 7 to
ring members containing 4 to 9 carbon atoms and 1 to 3 heteroatoms
selected from N, 0, P and S; wherein W5 is independently substituted with 0 to
3 R2 groups;
10 W6 is -R5, -W5, or -R5aW5,
each m1 is independently an integer from 0 to 2;
and the salts, solvates, resolved enantiomers and purified diasteriomers
thereof.
The present invention as claimed also provides a compound of formula
(XXII) or (XXlla):
Ws'~' N Et Ws,,,, N Et
(XXII) (XXlla)
T, T,
G, G1
wherein:
E1 is -(CR1 R1)m1 W1 ;
G1 is W2 or -NHR1, -C(NH)(NH2), -NR1-C(NR1)(NR1R3), -NH-C(NH)(NHR3),
-NH-C(NH)(NHR1), -NH-C(NH)NH2, -CH(CH2NHR1)(CH20H),
-CH(CH2NHR1)(CH2NHR1), -CH(NHR1)-(CR1 R1)m2-CH(NHR1 R1,
-CH(OH}(CR1)R1)m2-CH(NHR1)R1, or -CH(NHR1 )(CR1 R1)m2-CH(OH)R1,
6c
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-(CR1 R1)m2-S-C(NH)NH2, -N=C(NHR1)(R3), -N=C(SR1)N(R1)2,
-N(R1)C(NH)N(R1)C=N, or -N=C(NHR1)(R1);
T1 is -NR1 W3, or a monocycle having 3 to 7 ring members containing 2
to 6 carbon atoms and 1 to 3 heteroatoms selected from N, 0, P and S,
or a bicycle having 7 to 10 ring members containing 4 to 9 carbon atoms
and 1 to 3 heteroatoms selected from N, 0, P and S;
R1 is independently H or alkyl of 1 to 12 carbon atoms;
R2 is independently R3 or R4 wherein each R4 is independently
substituted with 0 to 3 R3 groups;
R3 is independently F, CI, Br, I, -CN, N3, -N02, -OR6a, -OR1, -N(R1)2,
-N(R1)(R6b), -N(R6b)2, -SR1, -SR6a, -S(O)R1, -S(0)2R1, -S(O)OR1, -S(O)OR6a,
-S(O)20R1, -S(O)2OR6a, -C(O)OR1, -C(O)R6c, -C(O)OR6a, -OC(O)R1,
-N(R1)(C(O)R1), -N(R6b)(C(O)R1), -N(R1)(C(O)OR1), -N(R6b)(C(O)OR1),
-C(O)N(R1)2, -C(O)N(R6b)(R1), -C(O)N(R6b)2, -C(NR1)(N(R1)2),
-C(N(R6b))(N(R1)2), -C(N(R1))(N(R1)(R6b)), -C(N(R6b))(N(R1)(R6b)),
-C(N(R1))(N(R6b)2), -C(N(R6b))(N(R6b)2), -N(R1)C(N(R1))(N(R1)2),
-N(R1)C(N(R1))(N(R1)(R6b)), -N(R1)C(N(R6b))(N(R1)2),
-N(R6b)C(N(R1))(N(R1)2). -N(R6b)C(N(R6b))(N(R1)2),
-N(R6b)C(N(R1))(N(R1)(R6b)2), -N(R1)C(N(R6b))(N(R1)(R6b)),
-N(R1)C(N(R1))(N(R6b)2), -N(R6b)C(N(R6b))(N(R1)(R6b)),
N(R6b)C(N(R1))(N(R6b)2), -N(R1)C(N(R6b))((N(R6b)2),
-N(R6b)C(N(R6b))(N(R6b)2), =0, =S, =N(R1), =N(R6b) or W5;
R3a is independently -CN, N3, -NO, -N02, -OR6a, -OR1, -N(R1)2,
-N(R1)(R6b), -N(R6b)2, -SR1, -SR6a, -S(O)R1, -S(O)2R1, -S(O)OR1,
-S(O)OR6a, -S(O)20R1, -S(O)2OR6a, -C(O)OR1, -C(O)R6c, -C(O)OR6a,
-OC(O)R1, -N(R1)(C(O)R1). -N(R6b)(C(O)R1), -N(R1)(C(O)0R1),
6d
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-N(Rgb)(C(O)ORj), -C(O)N(R1)2, -C(O)N(R6b)(R1), -C(O)N(R6b)2,
-C(NR1)(N(R1)2), -C(N(R6b))(N(R1)2), -C(N(R1))(N(R1)(R6b)),
-C(N(R6b))(N(R1)(R6b)), -C(N(R1))(N(R6b)2), -C(N(R6b))(N(R6b)2),
-N(R1)C(N(R1))(N(R1)2), -N(R1)C(N(R1))(N(R1)(R6b)), -N(R1)C(N(R6b))(N(R1)2),
-N(R6b)C(N(R1))(N(R1)2), -N(R6b)C(N(R6b)XN(R1)2), -N(R6b)C(N(R1))(N(R1 XR6b)),
-N(R1)C(N(R6b)XN(R1 XR6b)), -N(R1)C(N(R1)XN(R6b)2), -N(R6b)C((N(R6b))N(R1
XR6b)),
-N(R6b)C(N(R1)XN(R6b)2), -N(R1)C(N(R6b))(N(R6b)2) or
-N(R6b)C(N(R6b))N(R6b)2),
R4 is independently alkyl of 1 to 12 carbon atoms, alkenyl of 2 to 12
carbon atoms, alkenyl of 2 to 12 carbon atoms, or alkynyl of 2 to 12 carbon
atoms;
R5 is independently R4 wherein each R4 is substituted with 0 to 3 R3
groups;
R5a is independently alkylene of 1 to 12 carbon atoms, alkenylene of 2 to
12 carbon atoms, or alkynylene of 2-12 carbon atoms any one of which alkylene,
alkenylene or alkynylene is substituted with 0-3 R3 groups;
R6a is independently H, an ether-group or an ester group;
R6b is independently H, a protecting group for amino or the residue of a
carboxyl-containing compound;
R6c is independently H or the residue of an amino-containing compound;
Wl is a group comprising an acidic hydrogen;
W2 is a group comprising a N or S atom;
W3 is W4 or W5;
W4 is R5 or -C(O)R5, -C(O)W5, -S02R5, or -S02W5;
6e
CA 02268756 2006-03-21
W5 is carbocycle selected from a monocycle containing 3 to 7 carbon
atoms and a bicycle containing 7 to 12 carbon atoms, or a heterocycle selected
from a monocycle having 3 to 7 ring members containing 2 to 6 carbon atoms
and 1 to 3 heteroatoms selected from N, 0, P and S, and a bicycle having 7 to
ring members containing 4 to 9 carbon atoms and 1 to 3 heteroatoms
selected from N, 0, P and S; wherein W5 is independently substituted with 0 to
3 R2 groups;
W6 is -R5, -W5, or -R5aW5,
each m1 is independently an integer from 0 to 2;
10 and the salts, solvates, resolved enantiomers and purified diasteriomers
thereof.
In another embodiment of the invention a compound or composition of
the invention is provided that further comprises a pharmaceutically acceptable
carrier.
In another embodiment of the invention the activity of neuraminidase is
inhibited by a method comprising the step of treating a sample suspected of
containing neuraminidase with a compound or composition of the invention.
The invention also concerns a use of the aforementioned compounds for
inhibiting the activity of neuraminidase wherein the compounds are in a form
suitable for contacting a sample suspected of containing neuraminidase.
Another embodiment of the invention provides a method for inhibiting the
activity of neuraminidase comprising the step of contacting a sample suspected
of containing neuraminidase with the composition embodiments of the invention.
Detailed Description
Compositions of the Invention.
The compounds of this invention exclude compounds heretofore
known. However, as will be further apparent below, in other embodiments, it
is within the invention to use for antiviral purposes known compounds
heretofore only produced and used as intermediates in the preparation of
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antiviral compounds. With respect to the United States, the compounds or
compositions herein exclude compounds that are anticipated under 35 USC
102 or obvious under 35 USC 103. In particular, the claims herein shall be
construed as excluding the compounds which are anticipated by or not
possessing novelty over WO 96/26933 (September 6, 1996); Nishimura, Y. et
al., "J. Am. Chem. Soc." 110:7249-7250 (1988); and "Bull. Chem. Soc. Jpn."
65:978-986 (1992), disclose the total synthesis of Siastatin B. Nishimura, Y.
et al.,
"J. Antibiotics" 45(10):1662-1668 (1992); 46(2): 300-309 (1993); 46(12):1883-
1889
(1993); 47(1):101-107 (1994); "Nat. Prod. Lett." 1(1):39-44 (1992); and
Japanese
Patent Applications 92-287381 (October 26, 1992); 90-201437 (July 31, 1990);
88-
125020 (May 24, 1988) and 50046895 (April 25, 1975).
In a further embodiment, the compounds of this invention are those in
which W6 is not -CH2OH, -CH2OAc, or -CH2OCH2Ph.
In a further embodiment, the compounds of this invention are those in
which El is not -CH2OH, -CH2OTMS, or -CHO.
In a further embodiment, the compounds of this invention are those in
which W6 is not polyhydroxyalkane, especially -CH(OH)CH(OH)CH2OH. In a
further embodiment, W6 is a branched chain group R5 as described below or a
carbocycle which is substituted with at least one group R5.
Whenever a compound described herein is substituted with more than
one of the same designated group, e.g., "R1" or "R6a", then it will be
understood that the groups may be the same or different, i.e., each group is
independently selected.
"Heterocycle" as used herein includes by way of example and not
limitation these heterocycles described in Paquette, Leo A., "Principles of
Modern Heterocyclic Chemistry" (W.A. Benjamin, New York, 1968),
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 "J. Am. Chem.
Soc.",
82:5566 (1960).
Examples of heterocycles include by way of example and not limitation
pyridyl, 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, tetrahydrofuranyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl,
decahydroquinolinyl, octahydroisoquinolinyl, azocinyl, triazinyl, 6H-1,2,5-
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thiadiazinyl, 2H,6H-1,5,2-dithiazinyl, thienyl, thianthrenyl, pyranyl,
isobenzofuranyl, chromenyl, xanthenyl, phenoxathiinyl, 2H-pyrrolyl,
isothiazolyl, isoxazolyl, pyrazinyl, pyridazinyl, indolizinyl, isoindolyl, 3H-
indolyl, 1H-indazoly, purinyl, 4H-quinolizinyl, phthalazinyl, naphthyridinyl,
quinoxalinyl, quinazolinyl, cinnolinyl, pteridinyl, 4aH-carbazolyl,
carbazolyl,
0-carbolinyl, phenanthridinyl, acridinyl, pyrimidinyl, phenanthrolinyl,
phenazinyl, phenothiazinyl, furazanyl, phenoxazinyl, isochromanyl,
chromanyl, imidazolidinyl, imidazolinyl, pyrazolidinyl, pyrazolinyl,
piperazinyl, indolinyl, isoindolinyl, quinuclidinyl, morpholinyl,
oxazolidinyl,
benzotriazolyl, benzisoxazolyl, oxindolyl, benzoxazolinyl, and isatinoyl.
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
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, 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-pyrazinyl, 3-pyrazinyl, 5-pyrazinyl, 6-pyrazinyl, 2-thiazolyl,
4-
thiazolyl, or 5-thiazolyl.
By way of example and not limitation, nitrogen bonded heterocycles are
bonded at position 1 of an aziridine, azetidine, pyrrole, pyrrolidine, 2-
pyrroline, 3-pyrroline, imidazole, imidazolidine, 2-imidazoline, 3-
irinidazoline, pyrazole, pyrazoline, 2-pyrazoline, 3-pyrazoline, piperidine,
piperazine, indole, indoline, 1H-indazole, position 2 of a isoindole, or
.isoindoline, position 4 of a morpholine, and position 9 of a carbazole, or 30
carboline. Still more typically, nitrogen bonded heterocycles include 1-
aziridyl,
1-azetedyl, 1-pyrrolyl, 1-imidazolyl, 1-pyrazolyl, and 1-piperidinyl.
"Alkyl" as used herein, unless stated to the contrary, is C1-C12
hydrocarbon containing normal, secondary, tertiary or cyclic carbon atoms.
Examples are methyl (Me, -CH3), ethyl (Et, -CH2CH3), 1-propyl (n-Pr, n-propyl,
-CH2CH2CH3), 2-propyl (i-Pr, i-propyl, -CH(CH3)2), 1-butyl (n-Bu, n-butyl,
-CH2CH2CH2CH3), 2-methyl-l-propyl (i-Bu, i-butyl, -CH2CH(CH3)2),
2-butyl (s-Bu, s-butyl, -CH(CH3)CH2CH3), 2-methyl-2-propyl (t-Bu, t-butyl,
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-C(CH3)3), 1-pentyl (n-pentyl, -CH2CH2CH2CH2CH3), 2-pentyl
(-CH(CH3)CH2CH2CH3), 3-pentyl (-CH(CH2CH3)2), 2-methyl-2-butyl
(-C(CH3)2CH2CH3), 3-methyl-2-butyl (-CH(CH3)CH(CH3)2), 3-methyl-l-butyl
(-CH2CH2CH(CH3)2), 2-methyl-l-butyl (-CH2CH(CH3)CH2CH3), 1-hexyl
(-CH2CH2CH2CH2CH2CH3), 2-hexyl (-CH(CH3)CH2CH2CH2CH3), 3-hexyl
(-CH(CH2CH3)(CH2CH2CH3)), 2-methyl-2-pentyl (-C(CH3)2CH2CH2CH3),
3-methyl-2-pentyl (-CH(CH3)CH(CH3)CH2CH3), 4-methyl-2-pentyl
(-CH(CH3)CH2CH(CH3)2), 3-methyl-3-pentyl (-C(CH3)(CH2CH3)2),
2-methyl-3-pentyl (-CH(CH2CH3)CH(CH3)2), 2,3-dimethyl-2-butyl
(-C(CH3)2CH(CH3)2), 3,3-dimethyl-2-butyl (-CH(CH3)C(CH3)3). Examples of
alkyl groups appear in Table 2 as groups 2-5, 7, 9, and 100-399.
The compositions of the invention comprise compounds of either
formula:
Z2
J2 J2 Z ,N J1
W6~N E1 ~1 E1
I T, J1a T1 N J1a
J2 G 1 J2a J2 Z'
(IX) or (X)
In the typical embodiment, the compounds of Formula IX are chosen.
T1 and J1a are independently R1, Br, Cl, F, I, CN, N02 or N3, typically R1
or F, more typically H or F, more typically yet H.
J2 and J2a are independently H or R1, typically H.
One Z1 of Formula X is W6 and the other is G1.
Z2 of Formula X is H or W6, typically H.
El is -(CR1R1)m1W1.
Typically, Ri is H or alkyl of 1 to 12 carbon atoms, usually H or an alkyl
of 1 to 4 or 5 to 10 carbon atoms, still more typically, H or an alkyl of 1,
2, 3, 4, 5,
6, 7, 8, 9, 10, 11, or 12 carbon atoms, more typically yet, H or an alkyl of 1
to 3
carbon atoms selected from methyl, ethyl, n-propyl, and i-propyl. Most
typically Ri is H.
ml is an integer of 0 to 2, typically 0 or 1, most typically 0.
m2 is an integer of 0 to 1.
m3 is an integer of 1 to 3.
Wi is a group comprising an acidic hydrogen, a protected acidic group
or an R6c amide of the group comprising an acidic hydrogen which, within
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the context of the invention, means a group having a hydrogen atom that can
be removed by a base yielding an anion or its corresponding salt or solvate.
The general principles of acidity and basicity of organic materials are well
understood and are to be understood as defining Wi. They will not be
detailed here. However, a description appears in Streitwieser, A. and
Heathcock, C. H. "Introduction to Organic Chemistry, Second Edition"
(Macmillan, New York, 1981), pages 60-64. Generally, acidic groups of the
invention have pK values less than that of water, usually less than pK = 10,
typically less than pK = 8, and frequently less than pK = 6. They include
tetrazoles and the acids of carbon, sulfur, phosphorous and nitrogen,
typically
the carboxylic, sulfuric, sulfonic, sulfinic, phosphoric and phosphonic acids,
together with the R6c amides and R6b esters of those acids (R6c and R6b are
defined below). Exemplary Wi are -CO2H, -CO2R6a, -OSO3H, -S03H, -S02H,
-OP03H2, -PO3(R6a)2, -P03H2, -P03(H)(R6a), and -OPO3(R6a)2. El typically is
W1, and W1 typically is -CO2H, -CO2R6a, -CO2R4 or CO2R1, and most typically
is CO2R14 wherein R14 is normal or terminally secondary Cl-C6 alkyl.
Wi may also be a protected acidic group, which, within the context of
the invention means an acidic group as described above that has been
protected by one of the groups commonly used in the art for such groups and
are described below under R6a. More typically, protected W1 is -CO2R1,-
SO3R1, -S(O)OR1, -P(O)(OR1)2, -C(O)NHSO2R4, or -SO2NHC(O)-R4, wherein
R1 and R4 are defined above.
Most typically, El is selected from -C(O)O(CH2)bCH((CH2)cCH3)2 where
b=0to4,c=0to4,andb+c=lto4,orfromthegroupof
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, ,0 _ ,0 O
~--~ ~( ~4 CH3
OH O-CH3 , O-/
~ 0 O CH3
O~CHs ~ CH3 ~--~ CH
o~ 3
CH3
0 CH3 O~ ~O
~S, OH
H
I
~\ ZOH 0~ ,O-CH3 ~N~
P" p~ N N
OH ' S~ O-CH , and ~ N 11
3 S
Exemplary El groups are listed in Tables 3a through 3b.
G1 of Formula X is -OH, OR6a, or -(CR1R1)m1W2, G1 of Formula IX is
-N3, -CN, -OH, OR6a, -NO2 or -(CR1R1)m1W2, wherein R1 and ml are
defined above. Ordinarily, G1 of Formula (IX) is -(CR1R1)m1W2 and G1 of
Formula (X) is H.
W2 of Formula (X) is H or a group comprising a basic heteroatom, a
protected basic heteroatom or an R6b amide of the basic heteroatom. W2 of
Formula (IX) is a group comprising a basic heteroatom, a protected basic
heteroatom or an R6b amide of the basic heteroatom. W2 generally comprises
a basic heteroatom, which, within the context of the invention means an atom
other than carbon which is capable of protonation, typically by an acidic
hydrogen having an acidity in the range described above for W1. The basic
principles of basicity are described in Streitwieser and Heathcock (op. cit.)
and
provide meaning for the term basic heteroatom as will be understood by those
ordinarily skilled in the art. Generally, the basic heteroatoms employed in
the
compounds of the invention have pK values for the corresponding
protonated form that are in the range of values described above for W1. Basic
heteroatoms include the heteroatoms common in organic compounds which
have an un-shared, non-bonding, n-type, or the like, electron pair. By way of
example and not limitation, typical basic heteroatoms include the oxygen,
nitrogen, and sulfur atoms of groups such as alcohols, amines, amidines,
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guanidines, sulfides, and the like, frequently, amines, amidines and
guanidines. Ordinarily, W2 is amino or an amino alkyl (generally lower alkyl
C1 to C6) group such as aminomethyl, aminoethyl or aminopropyl; an
amidinyl, or an amidinoalkyl group such as amidinomethyl, amidinoethyl, or
amidinopropyl; or guanidinyl, or a guanidinoalkyl group such as
guanidinomethyl, guanidinoethyl, or guanidinopropyl (in each instance
wherein the alkyl group serves to bridge the basic substituent to the
carbocyclic
ring). More typically, W2 is amino, amidino, guanidino, heterocycle,
heterocycle substituted with 1 or 2 amino or guanidino groups (usually 1), or
an alkyl of 2 to 3 carbon atoms substituted with amino or guanidino, or such
alkyl substituted with an amino and a second group selected from the group
consisting of hydroxy and amino. The heterocycles useful as W2 include
typically N or S-containing 5 or 6 membered rings, wherein the ring contains 1
or 2 heteroatoms. Such heterocycles generally are substituted at ring carbon
atoms. They may be saturated or unsaturated and may be linked to the core
cyclohexene by lower alkyl (m1=1 or 2) or by -NR1-. Still more typically, W2
is
-NHR1, -C(NH)(NH2), -NR1-C(NR1)(NR1R3), -NH-C(NH)(NHR3), -NH-
C(NH)(NHR1),
-NH-C(NH)NH2, -CH(CH2NHR1)(CH2OH), -CH(CH2NHR1)(CH2NHR1),
-CH(NHR1)-(CR1R1)m2-CH(NHR1)R1, -CH(OH)-(CR1R1)m2-CH(NHR1)R1,
or -CH(NHR1)-(CR1R1)m2-CH(OH)R1, -(CR1R1)m2-S-C(NH)NH2,
-N=C(NHR1)(R3), -N=C(SR1)N(R1)2, -N(R1)C(NH)N(R1)C=N, or
-N=C(NHR1)(R1); wherein each m2 is ordinarily 0, and ordinarily R1 is H and
R3 is C(O)N(R1)2.
W2 optionally is a protected basic heteroatom which within the context
of the invention means a basic heteroatom as described above that has been
protected by R6b such as one of the groups common in the art. Such groups
are described in detail in Greene (op. cit.) as set forth below. Such groups
include by way of example and not limitation, amides, carbamates, amino
acetals, imines, enamines, N-alkyl or N-aryl phosphinyls, N-alkyl or N-aryl
sulfenyls or sulfonyls, N-alkyl or N-aryl silyls, thioethers, thioesters,
disulfides, sulfenyls, and the like. In some embodiments, the protecting group
R6b will be cleavable under physiological conditions, typically it will be
cleavable in vivo where, for example, the basic heteroatom forms an amide
with an organic acid or an amino acid such as a naturally occurring amino acid
or a polypeptide as described below for the R6a group.
Typically G1 of Formula (X) is H and G1 of Formula (IX) is selected from
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the group consisting of:
NH2 NH2 NH2 NH2
= ' V--~ NH ' 'OC~ NH
H H
I I
~~Ny NH ~NH2 ,/'~Ny NH
NH2 ' NH2
YNYNH2 YNYNH2 YNy NH2
H CH3 SCH3
H CH3 H ~CH3 H CH3
YN N YN N / N N
NH2
NH2 HNH2
CN ~ HN N, H IY N~H
N H2 S y ,N HN- CN H ,
NH2
H H H
I I I
I< N, CH3 NuCH3 "-'~CH
3
3
H H H
I I I
Ny CH3 OH NH2
CH3
= CH3 cCH3
N, CH3 and N'----CH3
Further exemplary G1 groups are listed in Table 4.
Ti is -NR1W3, -R3, -R5 or heterocycle. Typically Ti is -NR1W3 or
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heterocycle. Generally T1 is selected from the group consisting of:
~ O O
H3C N FH2CN HF2CJ~NA
H H H
O
O O
N N
F3C N~ N N ~
~ and
H > > > .
O
Exemplary Ti groups are listed in Table 5.
W3 is W4 or W5, wherein W4 is R5 or -C(O)R5, -C(O)W5, -S02R5, or
-S02W5_ Typically, W3 is -C(O)R5 or W5.
R2 is independently R3 or R4 as defined below, with the proviso that
each R4 is independently substituted with 0 to 3 R3 groups;
R3 is independently F, Cl, Br, I, -CN, N3, -NO2, -ORba, -ORl, -N(Rl)2,
-N(R1)(R6b), -N(R6b)2, -SR1, -SR6a, -S(O)R1. -S(0)2R1, -S(O)OR1, -S(O)ORba.
-S(O)20R1, -S(0)2OR6a, -C(O)ORl, -C(O)R6c, -C(O)OR6a, -OC(O)Rl,
-N(Rl)(C(O)R1), -N(R6b)(C(O)R1), -N(R1)(C(O)ORl), -N(R6b)(C(O)OR1),
-C(O)N(Rl)2, -C(O)N(R6b)(Rl), -C(O)N(R6b)2, -C(NR1)(N(R1)2),
-C(N(R6b))(N(Rl)2), -C(N(R1))(N(R1)(R6b)), -C(N(R6b))(N(Rl)(R6b)),
-C(N(Rl))(N(R6b)2), -C(N(R6b))(N(R6b)2), -N(R1)C(N(Rl))(N(R1)2),
-N(Rl)C(N(Rl))(N(R1)(R6b)), -N(Rl)C(N(R6b))(N(Rl)2),
-N(R6b)C(N(Rl))(N(Rl)2), -N(R6b)C(N(R6b))(N(R1)2),
-N(R6b)C(N(Rl))(N(Rl)(R6b)), -N(Rl)C(N(R6b))(N(Rl)(R6b)),
-N(Rl)C(N(R1))(N(R6b)2), -N(R6b)C(N(R6b))(N(R1)(R6b)),
-N(R6b)C(N(Rl))(N(R6b)2), -N(Rl)C(N(R6b))(N(R6b)2),
-N(R6b)C(N(R6b))(N(R6b)2), =0, =S, =N(Ri), =N(R6b) or W5. Typically R3 is F,
Cl, -CN, N3, N02, -OR6a, -OR1, -N(R1)2, -N(R1)(R6b), -N(R6b)2, -SR1, -SR6a,
-C(O)OR1. -C(O)R6c, -C(O)OR6a, -OC(O)R1, -NR1C(O)R1, -N(R6b)C(O)R1,
-C(O)N(R1)2, -C(O)N(R6b)(R1), -C(O)N(R6b)2, or =0. More typical R3 groups
comprising R6b include -C(O)N(R6b)2 or -C(O)N(R6b)(R1). More typically yet
R3 is F, Cl, -CN, N3, -OR1, -N(R1)2, -SR1, -C(O)OR1, -OC(O)R1, or =0. More
typically still, R3 is F, -OR1, -N(R1)2, or =0. In the context of the present
application, "=0" denotes a double bonded oxygen atom (oxo), and "=S"
=N(R6b) and "=N(R1)" denote the sulfur and nitrogen analogs.
R4 is alkyl of 1 to 12 carbon atoms, and alkynyl or alkenyl of 2 to 12
carbon atoms. The alkyl R4's are typically of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,
11, or 12
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carbon atoms and the alkenyl and alkynyl R4's are typically of 2, 3, 4, 5, 6,
7, 8,
9, 10, 11, or 12 carbon atoms. R4 ordinarily is alkyl (as defined above). When
R4 is alkenyl it is typically ethenyl (-CH=CH2), 1-prop-l-enyl (-CH=CHCH3), 1-
prop-2-enyl (-CH2CH=CH2), 2-prop-1-enyl (-C(=CH2)(CH3)), 1-but-l-enyl
(-CH=CHCH2CH3), 1-but-2-enyl (-CH2CH=CHCH3), 1-but-3-enyl
(-CH2CH2CH=CH2), 2-methyl-l-prop-l-enyl (-CH=C(CH3)2), 2-methyl-l-prop-
2-enyl (-CH2C(=CH2)(CH3)), 2-but-1-enyl (-C(=CH2)CH2CH3), 2-but-2-enyl
(-C(CH3)=CHCH3), 2-but-3-enyl (-CH(CH3)CH=CH2), 1-pent-l-enyl
(-C=CHCH2CH2CH3), 1-pent-2-enyl (-CHCH=CHCH2CH3), 1-pent-3-enyl
(-CHCH2CH=CHCH3), 1-pent-4-enyl (-CHCH2CH2CH=CH2), 2-pent-1-enyl
(-C(=CH2)CH2CH2CH3), 2-pent-2-enyl (-C(CH3)=CH2CH2CH3), 2-pent-3-enyl
(-CH(CH3)CH=CHCH3), 2-pent-4-enyl (-CH(CH3)CH2CH=CH2) or 3-methyl-l-
but-2-enyl (-CH2CH=C(CH3)2). More typically, R4 alkenyl groups are of 2, 3 or
4 carbon atoms. When R4 is alkynyl it is typically ethynyl (-C=CH), 1-prop-l-
ynyl (-C=CCH3),1-prop-2-ynyl (-CH2C=CH), 1-but-1-ynyl (-C=CCH2CH3), 1-but-
2-ynyl (-CH2C=CCH3),1-but-3-ynyl (-CH2CH2C=CH), 2-but-3-ynyl
(CH(CH3)C=CH), 1-pent-1-ynyl (-C=CCH2CH2CH3), 1-pent-2-ynyl
(-CH2C-CCH2CH3),1-pent-3-ynyl (-CH2CH2C=_CCH3) or 1-pent-4-ynyl
(-CH2CH2CH2C=CH). More typically, R4 alkynyl groups are of 2, 3 or 4 carbon
atoms.
R5 is R4, as defined above, or R4 substituted with 0 to 3 R3 groups.
Typically R5 is an alkyl of 1 to 4 carbon atoms substituted with 0 to 3
fluorine
atoms.
R5a is independently alkylene of 1 to 12 carbon atoms, alkenylene of 2 to
12 carbon atoms, or alkynylene of 2-12 carbon atoms any one of which
alkylene, alkenylene or alkynylene is substituted with 0-3 R3 groups. As
defined above for R4, R5a's are of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12
carbon atoms
when alkylene and of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 carbon atoms when
alkenylene or alkynylene. Each of the typical R4 groups is a typical R5a group
with the proviso that one of the hydrogen atoms of the described R4 group is
removed to form the open valence to a carbon atom through which the
second bond to the R5a is attached.
R14 is normal or terminally secondary C1-C6 alkyl.
W5 is a carbocycle or heterocycle, with the proviso that each W5 is
independently substituted with 0 to 3 R2 groups. W5 carbocycles and T1 and
W5 heterocycles are stable chemical structures. Such structures are isolatable
in measurable yield, with measurable purity, from reaction mixtures at
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temperatures from -78 C to 200 C. Each W5 is independently substituted with
0 to 3 R2 groups. Typically, T1 and W5 are a saturated, unsaturated or
aromatic ring comprising a mono- or bicyclic carbocycle or heterocycle. More
typically, T1 or W5 has 3 to 10 ring atoms, still more typically, 3 to 7 ring
atoms, and ordinarily 3 to 6 ring atoms. The T1 and W5 rings are saturated
when containing 3 ring atoms, saturated or monounsaturated when
containing 4 ring atoms, saturated, or mono- or diunsaturated when
containing 5 ring atoms, and saturated, mono- or diunsaturated, or aromatic
when containing 6 ring atoms. Unsaturation of the W5 rings include internal
and external unsaturation wherein the external incorporates a ring atom.
When W5 is carbocyclic, it is typically a 3 to 7 carbon monocycle or a 7 to
12 carbon atom bicycle. More typically, W5 monocyclic carbocycles have 3 to 6
ring atoms, still more typically 5 or 6 ring atoms. W5 bicyclic carbocycles
typically have 7 to 12 ring atoms arranged as a bicyclo [4,5], [5,5], [5,6] or
[6,6]
system, still more typically, 9 or 10 ring atoms arranged as a bicyclo [5,6]
or [6,6]
system. Examples include cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-l-
enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclohexyl, 1-cyclohex-l-enyl, 1-
cyclohex-2-enyl, 1-cyclohex-3-enyl, phenyl, spiryl and naphthyl.
A T1 or W5 heterocycle is typically a monocycle having 3 to 7 ring
members (2 to 6 carbon atoms and 1 to 3 heteroatoms selected from N, 0, 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, 0, P, and S). More typically, T1 and W5
heterocyclic monocycles have 3 to 6 ring atoms (2 to 5 carbon atoms and 1 to 2
heteroatoms selected from N, 0, and S), still more typically, 5 or 6 ring
atoms
(3 to 5 carbon atoms and 1 to 2 heteroatoms selected from N and S). T1 and
W5 heterocyclic bicycles have 7 to 10 ring atoms (6 to 9 carbon atoms and 1 to
2
heteroatoms selected from N, 0, and S) arranged as a bicyclo [4,5], [5,5],
[5,6], or
[6,6] system, still more typically, 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.
Typically Ti and W5 heterocycles are selected from pyridyl, pyridazinyl,
pyrimidinyl, pyrazinyl, s-triazinyl, oxazolyl, imidazolyl, thiazolyl,
isoxazolyl,
pyrazolyl, isothiazolyl, furanyl, thiofuranyl, thienyl, or pyrrolyl.
More typically, the heterocycle of T1 and W5 is bonded through a carbon
atom or nitrogen atom thereof. Still more typically Ti heterocycles are bonded
by a stable covalent bond through a nitrogen atom thereof to the cyclohexene
ring of the compositions of the invention and W5 heterocycles are bonded by a
stable covalent bond through a carbon or nitrogen atom thereof to the
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cyclohexene ring of the compositions of the invention. Stable covalent bonds
are chemically stable structures as described above.
W5 optionally is selected from the group consisting of:
N
a'N f
H
N N ~
~ N
N H,
S'Y~ <Nl N~~
/
N S and \~-S
W6 is -R5, -W5, -R5aW5, -C(O)OR6a, -C(O)R6c. -C(O)N(R6b)2,
-C(NR6b)(N(R6b)2), -C(NR6b)(N(H)(R6b)), -C(N(H)(N(R6b)2), -C(S)N(R6b)2, or
-C(O)R2, typically W6 is -R5, -W5, or -R5aW5; in some embodiments, W6 is
R1, -C(O)-R1, -CHR1W7, -CH(R1)aW7, -CH(W7)2, (where, W7 is monovalent a
is 0 or 1, but is 0 when W7 is divalent) or -C(O)W7. In some embodiments,
W6 is -CHR1W7 or -C(O)W7, or W6 is -(CH2)m1CH((CH2)m3R3)2;
-(CH2)m1C((CH2)m3R3)3; -(CH2)m1CH((CH2)m3R5aW5)2;
-(CH2)m1CH((CH2)m3R3)((CH2)m3R5aW5)%
-(CH2)mlC((CH2)m3R3)2(CH2)m3R5aW5); -(CH2)mlC((CH2)m3R5aW5)3 or
-(CH2)m1C((CH2)m3R3)((CH2)m3R5aW5)2; and wherein m3 is an integer
from 1 to 3.
W7 is R3 or R5, but typically is alkyl of 1 to 12 carbons substituted with 0
to 3 R3 groups, the latter typically selected from the group consisting of
-NR1(R6b), -N(R6b)2, -OR6a, or SR6a. More typically, W7 is -OR1 or an alkyl
of 3 to 12 carbon atoms substituted with ORl.
In general, W6 is R1-, -CHR1 W 7,
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OH OH
HO"'~
OH OH
H3C
CH3
CH3 CH3
CH3
H3C CH3 CH3 , and
Exemplary W6 groups are listed in Table 2.
An embodiment of the invention comprises a compound of the
formula:
R1
W6'~, N W1 W6_ N'
, (XIII)
W3", (XI) W
W3
R1 /N' N N
W3 R, or R1 R6b
wherein each R1 and R6b are typically H, and W2 is typically selected from the
group consisting of:
O 0 O
O~S-
OH + O- Rsa OH ,
H
I
O\ /OH O\\ O-R6a ~N
P" OH yP~O_R , and N N
6a
and W6 is one of:
R~
R7-/\ f R7/~~~ f R7~
I R7 ' R7 , and
wherein R7 is H, -CH3, -CH2CH3, -CH2CH2CH3, -OCH3, -OAc (-O-C(O)CH3),
-OH, -NH2, or -SH, typically H, -CH3 or -CH2CH3. .
Another embodiment of the invention is directed toward compounds
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of formula (XX) or (XXa):
G2 J1 G2 J2 J2a
J3 E1 E1
(XX) J3 (XXa)
T1 A3 J2a T1 A3 j 1
J2 1 ~2 J2 I
Z3 Z3
wherein
A3 is N, N(O) or N(S);
Z3 is H, W6, G1 or R3a;
El is -(CR1R1)m1W1;
G1 is N3, -CN, -OH, -OR6a, -N02 or -(CR1R1)m1W2;
G2 is G1 or -X1W6;
T1 is -NR1W3 or a heterocycle;
U. is R1, Br, Cl, F, I, CN, N02 or N3;
J2 is H or R1;
J3 is J1 if X1 is a bond; and J3 is J2 if X1 is -0-, -N(H)-, -N(W6)-, -N(OH)-,
-N(OW6)-, -N(NH2)-, -N(N(H)(W6))-, -N(N(W6)2)-, -N(H)N(W6)-, -S-, -SO-, or
-S02-;
R1 is independently H or alkyl of 1 to 12 carbon atoms;
R2 is independently R3 or R4 wherein each R4 is independently
substituted with 0 to 3 R3 groups;
R3 is independently F, Cl, Br, I, -CN, N3, -N02, -OR6a, -OR1, -N(R1)2,
-N(R1)(R6b), -N(R6b)2, -SR1, -SR6a, -S(O)R1, -S(O)2R1, -S(O)OR1, -S(O)OR6a,
-S(O)2OR1, -S(O)2OR6a, -C(O)OR1, -C(O)R6c, -C(O)ORba, -OC(O)R1,
-N(R1)(C(O)R1), -N(R6b)(C(O)R1), -N(R1)(C(O)0R1), -N(R6b)(C(O)OR1),
-C(O)N(R1)2, -C(O)N(R6b)(R1), -C(O)N(R6b)2, -C(NR1)(N(R1)2),
-C(N(R6b))(N(R1)2), -C(N(R1))(N(R1)(R6b)), -C(N(R6b))(N(R1)(R6b)),
-C(N(R1))(N(R6b)2), -C(N(R6b))(N(R6b)2), -N(Rl)C(N(Rl))(N(Rl)2),
-N(R1)C(N(R1))(N(R1)(R6b)), -N(R1)C(N(R6b))(N(R1)2),
-N(R6b)C(N(R1))(N(R1)2), -N(R6b)C(N(R6b))(N(R1)2),
-N(R6b)C(N(R1))(N(R1)(R6b)), -N(R1)C(N(R6b))(N(R1)(R6b))'
-N(R1)C(N(R1))(N(R6b)2), -N(R6b)C(N(R6b))(N(R1)(R6b)).
-N(R6b)C(N(R1))(N(R6b)2), -N(R1)C(N(R6b))(N(R6b)2),
-N(R6b)C(N(R6b))(N(R6b)2), =0, =S, =N(R1). =N(R6b) or W5;
R3a is independently -CN, N3, -NO, -N02, -OR6a, -OR1, -N(R1)2,
-N(R1)(R6b), -N(R6b)2, -SR1, -SR6a, -S(O)R1, -S(O)2R1, -S(O)OR1, -S(O)OR6a,
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-S(O)2OR1, -S(O)2OR6a, -C(O)OR1, -C(O)R6c, -C(O)OR6a, -OC(O)Rl,
-N(R1)(C(O)Rl), -N(R6b)(C(O)Rl), -N(R1)(C(O)OR1), -N(R6b)(C(O)ORl),
-C(O)N(RI)2, -C(O)N(R6b)(RZ), -C(O)N(R6b)2, -C(NR1)(N(Rl)2).
-C(N(R6b))(N(Rl)2), -C(N(R1))(N(RI)(R6b)), -C(N(R6b))(N(R1)(R6b)).
-C(N(RZ))(N(R6b)2), -C(N(R6b))(N(R6b)2), -N(R1)C(N(Ri))(N(Rl)2),
-N(R1)C(N(Ri))(N(RZ)(R6b)), -N(Rl)C(N(R6b))(N(R7)2),
-N(R6b)C(N(Rj))(N(R1)2), -N(R6b)C(N(R6b))(N(Rl)2).
-N(R6b)C(N(R1))(N(Rl)(R6b)), -N(Rl)C(N(R6b))(N(Rl)(R6b)),
-N(R1)C(N(Ri))(N(R6b)2), -N(R6b)C(N(R6b))(N(Rl)(R6b)),
-N(R6b)C(N(R1))(N(R6b)2), -N(R1)C(N(R6b))(N(R6b)2) or
-N(R6b)C(N(R6b))(N(R6b)2);
R4 is independently alkyl of 1 to 12 carbon atoms, alkenyl of 2 to 12
carbon atoms, or alkynyl of 2 to 12 carbon atoms;
R5 is independently R4 wherein each R4 is substituted with 0 to 3 R3
groups;
R5a is independently alkylene of 1 to 12 carbon atoms, alkenylene of 2 to
12 carbon atoms, or alkynylene of 2-12 carbon atoms any one of which
alkylene, alkenylene or alkynylene is substituted with 0-3 R3 groups;
R6a is independently H or an ether- or ester-forming group;
R6b is independently H, a protecting group for amino or the residue of a
carboxyl-containing compound;
R6c is independently H or the residue of an amino-containing
compound;
Wi is a group comprising an acidic hydrogen, a protected acidic group,
or an R6c amide of the group comprising an acidic hydrogen;
W2 is a group comprising a basic heteroatom or a protected basic
heteroatom, or an R6b amide of the basic heteroatom;
W3 is W4 or W5;
W4 is R5 or -C(O)R5, -C(O)W5, -S02R5, or -S02W5;
W5 is carbocycle or heterocycle wherein W5 is independently
substituted with 0 to 3 R2 groups;
W6 is -R5, -W5, -R5aW5, -C(O)OR6a, -C(O)R6c, -C(O)N(R6b)2.
-C(NR6b)(N(R6b)2), -C(NR6b)(N(H)(R6b)), -C(N(H)(N(R6b)2), -C(S)N(R6b)2, or
-C(O)R2;
X1 is a bond, -0-, -N(H)-, -N(W6)-, -N(OH)-, -N(OW6)-, -N(NH2)-,
-N(N(H)(W6))-, -N(N(W6)2)-, -N(H)N(W6)-, -S-, -SO-, or -S02-; and
each m1 is independently an integer from 0 to 2;
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provided, however, that compounds of formula (XX) are excluded wherein A3
is N, each J1, J2, J2a and J3 is H and Tl is -N(H)(Ac) and:
E1 is -CO2H or -CO2CH3,
G2 is -OBoc, and
Z3 is Boc;
E1 is -CO2H or -CO2CH3,
G2 is -OH, and
Z3 is H;
E1 is -CO2H, -CO2CH3 or -CO2Bn
G2 is -OH, and
Z3 is Boc;
El is -CONH2,
G2 is -OH, and
Z3 is Boc or H;
El is -CO2H or -CO2CH3,
G2 is OH, and
Z3 is Bn; or
El is -CO2H or -CO2CH3,
G2 is -OH, and
Z3 is -CH2CH(OH)CH2(OH);
wherein Bn is benzyl and Boc is -CO2C(CH3)3;
further excluded are compounds of the formula: (VII) or (VIII):
U1"'~ U1-~'
N E1 N E1
T1 ,J1a (VII) T1 J (VIII)
1a
J2 J1 J2
G1 J2a 1 J2a
wherein
El is -(CR1R1)m1W1;
Gi is N3, -CN, -OH, -OR6a, -NO2, or -(CR1R1)m1W2;
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T1 is -NR1W3, a heterocycle, or is taken together with G1 to form a
group having the structure
R6b-N
;
U1 is -X1W6;
J1 and J1a are independently R1, Br, Cl, F, I, CN, N02 or N3;
J2 and J2a are independently H or R1;
R1 is independently H or alkyl of 1 to 12 carbon atoms;
R2 is independently R3 or R4 wherein each R4 is independently
substituted with 0 to 3 R3 groups;
R3 is independently F, Cl, Br, I, -CN, N3, -NO2, -OR6a, -OR1, -N(R1)2,
-N(R1)(R6b), -N(R6b)2, -SR1, -SR6a, -S(O).R1. -S(O)2R1, -S(O)OR1, -S(O)OR6a,
-S(O)20R1, -S(O)2OR6a, -C(O)OR1, -C(O)R6c, -C(O)OR6a, -OC(O)Rl,
-N(Rl)(C(O)R1), -N(R6b)(C(O)Rl), -N(R1)(C(O)ORl), -N(R6b)(C(O)OR1).
-C(O)N(R1)2, -C(O)N(R6b)(R1), -C(O)N(R6b)2, -C(NR1)(N(Rl)2),
-C(N(R6b))(N(Rl)2), -C(N(Rl))(N(R1)(R6b)), -C(N(R6b))(N(R1)(R6b)),
-C(N(Rl))(N(R6b)2), -C(N(R6b))(N(R6b)2), -N(R1)C(N(RI))(N(Rl)2),
-N(Rl)C(N(R1))(N(Ri)(R6b)), -N(RI)C(N(R6b))(N(Rl)2),
-N(R6b)C(N(Rl))(N(RZ)2), -N(R6b)C(N(R6b))(N(R1)2),
-N(R6b)C(N(RZ))(N(RZ)(R6b)), -N(Rl)C(N(R6b))(N(Ri)(R6b)),
-N(Rl)C(N(RZ))(N(R6b)2), -N(R6b)C(N(R6b))(N(Rl)(R6b)).
-N(R6b)C(N(Rl))(N(R6b)2), -N(Ri)C(N(R6b))(N(R6b)2),
-N(R6b)C(N(R6b))(N(R6b)2), =0, =S, =N(Rl) or =N(R6b);
R4 is independently alkyl of 1 to 12 carbon atoms, alkenyl of 2 to 12
carbon atoms, or alkynyl of 2 to 12 carbon atoms;
R5 is independently R4 wherein each R4 is substituted with 0 to 3 R3
groups;
R5a is independently alkylene of 1 to 12 carbon atoms, alkenylene of 2 to
12 carbon atoms, or alkynylene of 2-12 carbon atoms any one of which
alkylene, alkenylene or alkynylene is substituted with 0-3 R3 groups;
R6a is independently H or an ether- or ester-forming group;
R6b is independently H, a protecting group for amino or the residue of a
carboxyl-containing compound;
R6c is independently H or the residue of an amino-containing
compound;
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W1 is a group comprising an acidic hydrogen, a protected acidic group,
or an R6c amide of the group comprising an acidic hydrogen;
W2 is a group comprising a basic heteroatom or a protected basic
heteroatom, or an R6b amide of the basic heteroatom;
W 3 is W4 or W5;
W4 is R5 or -C(O)R5, -C(O)W5, -S02R5, or -S02W5;
W5 is carbocycle or heterocycle wherein W5 is independently
substituted with 0 to 3 R2 groups;
W6 is -R5, -W5, -R5aW5, -C(O)OR6a, -C(O)R6c, -C(O)N(R6b)2,
-C(NR6b)(N(R6b)2), -C(NR6b)(N(H)(R6b)), -C(N(H)(N(R6b)2), -C(S)N(R6b)2, or
-C(O)R2;
X1 is a bond, -0-, -N(H)-, -N(W6)-, -S-, -SO-, or -S02-; and
each ml is independently an integer from 0 to 2;
and the salts, solvates, resolved enantiomers and purified diastereomers
thereof.
Typically A3 is N or N(O), more typically A3 is N.
In typical embodiments, one of Z3 and G2 is G1 or R3a and the other is
W6 or -X1W6. More typically, Z3 is W6 or R3a and G2 is G1 or -X1W6. More
typically yet, Z3 is W6 and G2 is G1; or Z3 is R3a and G2 is -X1W6.
J3 is Jl if Xl is a bond; and J3 is J2 if X1 is -0-, -N(H)-, -N(W6)-, -N(OH)-,
-N(OW6)-, -N(NH2)-, -N(N(H)(W6))-, -N(N(W6)2)-, -N(H)N(W6)-, -S-, -SO-, or
-S02-. The typical embodiments of J1 and J2, described above, are typical
embodiments of J3.
An embodiment of the invention comprises a compound of the
formula (XXI) or (XXIa):
J2 J2a J1
W6\N E1 W6~N E1
T I (XXI) (XXIa)
1 J1a Ti J2a
J2 G1 J2 J2 G1 2 An An embodiment of the invention comprises a compound of the
= formula (XXII) or (XXIIa):
W6 E1 ws\N E1
~ (XXII) (XXlla)
T1 T1
G1 G1
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An embodiment of the invention comprises a compound of the
formula (XXIII) or (XXIIIa):
Ws~N Wi Ws~N W1
W3~ I (XXIII) Ws~ (XXllla)
N N
I I =
R1 H~N\ R R1 H~N\ R
1 1
An embodiment of the invention comprises a compound of the
formula (XXIV) or (XXIVa):
N Z2 J1 Z2 J2 J2a
Z - J2 E1 Z~2-N E1
T J2a (XXIV) T (XXIVa)
1 N 1 N J1
J2 ~1 J2 J2 ~1
wherein one Z1 is W6 and the other Z1 is Gl; and Z2 is H or W6. Typically, Z2
is H.
An embodiment of the invention comprises a compound of the
formula (XXV) or (XXVa):
Z2 Z2
Ws N J1 W6 N J2 J2a
J2 E1 J2 E1
T J2a (XXV) T ~ (XXVa)
1 N 1 N J1
J2 G1 J2 J2 G1
An embodiment of the invention comprises a compound of the
formula (XXVI) or (XXVIa):
R1 R1
Ws N''1.. E1 W6 N'~~.. E1
(XXVI) ! (XXVIa)
Ti N Ti N
6, 61
An embodiment of the invention comprises a compound of the
formula (XXVII) or (XXVIIa):
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R 1 R 1
Ws N,,, W i Ws NW i
(XXVII) (XXVIIa)
W3' N W3~ N
R1 H,N~R, R, HRi
An embodiment of the invention comprises a compound of the
formula (XXVIII) or (XXVIIIa):
Ws O J1 Ws O J2 J2a
J2 \ E1 J2 E1
(XXVIII) I (XXVllla)
TIN J2a T1 N J1
J2 1 J2 J2
G1 G1
An embodiment of the invention comprises a compound of the
formula (XXIX) or (XXIXa):
W6 0/,,.,, El Ws 0Ei
(XXIX) I (XXIXa)
T, N T,
N
G1 G1
An embodiment of the invention comprises a compound of the
formula (XXX) or (XXXa):
Wg-O/i=... W1 Wfi O/i=.. W1
Ws',(XXX) W3~ I (XXXa)
N N N N
i ( Ri I
R1 H'N\R1 1 H~N\R,
It is important to appreciate that each of the typical embodiments of
formula (XXX) and (XXXa), set forth above and in the appended claims, are
also embodiments of formulas (XXI)-(XXX) and (XXIa)-(XXXa), set forth
immediately above.
Groups R6a and R6b are not critical functionalities and may vary widely.
When not H, their function is to serve as intermediates for the parental drug
substance. This does not mean that they are biologically inactive. On the
contrary, a principal function of these groups is to convert the parental drug
into a prodrug, whereby the parental drug is released upon conversion of the
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prodrug in vivo. Because active prodrugs are absorbed more effectively than
the parental drug they in fact often possess greater potency in vivo than the
parental drug. When not hydrogen, R6a and R6b are removed either in vitro,
in the instance of chemical intermediates, or in vivo, in the case of
prodrugs.
With chemical intermediates, it is not particularly important that the
resulting pro-functionality products, e.g. alcohols, be physiologically
acceptable, although in general it is more desirable if the products are
pharmacologically innocuous.
R6a is H or an ether- or ester-forming group. "Ether-forming group"
means a group which is capable of forming a stable, covalent bond between the
parental molecule and a group having the formula:
S-0-Va(V1)3 ~ S-0-Va(V1)(U2) ~ S-0-Va(V3)
S-O-ub(V1)2 S-0-Ub(V2) , or S-O-UcM)
Wherein Va is a tetravalent atom typically selected from C and Si; Vb is a
trivalent atom typically selected from B, Al, N, and P, more typically N and
P;
Vc is a divalent atom typically selected from 0, S, and Se, more typically S;
Vi
is a group bonded to Va, Vb or Vc by a stable, single covalent bond, typically
Vi
is W6 groups, more typically Vi is H, R2, W5, or -R5aW5, still more typically
H or R2; V2 is a group bonded to Va or Vb by a stable, double covalent bond,
provided that V2 is not =0, =S or =N-, typically V2 is =C(V1)2 wherein V1 is
as
described above; and V3 is a group bonded to Va by a stable, triple covalent
bond, typically V3 is -C(V1) wherein V1 is as described above.
. "Ester-forming group" means a group which is capable of forming a
stable, covalent bond between the parental molecule and a group having the
formula:
S-O-Va(V1)N4) S-0-Ub(U4) S-0-Ud(V1)2(V4)
S-0-Ud(V4)2 9 S O-Ve(V1)3(V4) , or S 0-Ve(U1)(V4)2
Wherein Va, Vb, and V1, are as described above; Vd is a pentavalent atom
typically selected from P and N; Ve is a hexavalent atom typically S; and V4
is
a group bonded to Va, Vb, Vd or Ve by a stable, double covalent bond,
provided that at least one V4 is =0, =S or =N-V1, typically V4, when other
than =0, =S or =N-, is =C(V1)2 wherein V1 is as described above.
Protecting groups for -OH functions (whether hydroxy, acid or other
functions) are embodiments of "ether- or ester-forming groups".
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Particularly of interest are ether- or ester-forming groups that are
capable of functioning as protecting groups in the synthetic schemes set forth
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 under R6c below. R6c is capable of
protecting hydroxyl or thio groups such that hydrolysis from the parental
molecule yields hydroxyl or thio.
In its ester-forming role, R6a 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 -CO2R6a. R6a for example is deduced from the enumerated ester
groups of WO 95/07920.
Examples of R6a include
C3-C12 heterocycle (described above) or C6-C12 aryl. These
aromatic groups optionally are polycyclic or monocyclic. Examples include
phenyl, spiryl, 2- and 3-pyrrolyl, 2- and 3-thienyl, 2- and 4-imidazolyl, 2-,
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-, 4-
and 5-
pyrimidinyl,
C3-C12 heterocycle or C6-C12 aryl substituted with halo, Ri, R1-O-
C1-C12 alkylene, C1-C12 alkoxy, CN, NO2, OH, carboxy, carboxyester, thiol,
thioester, C1-C12 haloalkyl (1-6 halogen atoms), C2-C12 alkenyl or C2-C12
alkynyl. Such groups include 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-
hydroxyphenyl, 2-, 3- and 4-0-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-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-trichloromethylphenyl), 4-N-methylpiperidinyl, 3-N-
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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 [C6-C9 aryl (including
phenoxy ethyl)), 2,2'-dihydroxybiphenyl, 2-, 3- and 4-N,N-
dialkylaminophenol, -C6H4CH2-N(CH3)2, trimethoxybenzyl, triethoxybenzyl,
J-j N/
\ I -CH2-O-C(O) N
2-alkyl pyridinyl (C1-4 alkyl); 0 H ; -~
R10(O)C
C4 - C8 esters of 2-carboxyphenyl; and Cl-C4 alkylene-C3-C6
aryl (including benzyl, -CH2-pyrrolyl, -CH2-thienyl, -CH2-imidazolyl, -CH2-
oxazolyl, -CH2-isoxazolyl, -CH2-thiazolyl, -CH2-isothiazolyl, -CH2-pyrazolyl,
-CH2-pyridinyl and -CH2-pyrimidinyl) substituted in the aryl moiety by 3 to 5
halogen atoms or 1 to 2 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 -CH2-CC13), C1-C12 alkyl (including methyl and
ethyl), C2-C12 alkenyl or C2-C12 alkynyl;
alkoxy ethyl [C2-C6 alkyl including -CH2-CH2-O-CH3 (methoxy
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(CH3)2, -C(CH3)3,
-CH2CH3, -(CH2)2CH3, -(CH2)3CH3, -(CH2)4CH3, -(CH2)5CH3, -CH2CH2F,
-CH2CH2C1, -CH2CF3, and -CH2CC13);
N O
'-/ ; -N-2-propylmorpholino, 2,3-dihydro-6-
hydroxyindene, sesamol, catechol monoester, -CH2-C(O)-N(R1)2, -CH2-
S(O)(Rl), -CH2-S(O)2(R1), -CH2-CH(OC(O)CH2R1)-CH2(OC(O)CH2Rl),
cholesteryl, enolpyruvate (HOOC-C(=CH2)-), glycerol;
a 5 or 6 carbon monosaccharide, disaccharide or oligosaccharide (3 to
9 monosaccharide residues);
triglycerides such as a-D-0-diglycerides (wherein the fatty acids
composing glyceride lipids generally are naturally occurring saturated or
unsaturated C6-26, C6-18 or C6-1o fatty acids such as linoleic, lauric,
myristic,
palmitic, stearic, oleic, palmitoleic, linolenic and the like fatty acids)
linked to
acyl of the parental compounds herein through a glyceryl oxygen of the
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WO 98/17647 PCT/US97/18193
triglyceride;
phospholipids linked to the carboxyl group through the phosphate
of the phospholipid;
phthalidyl (shown in Fig. 1 of Clayton et al., "Antimicrob. Agents
Chemo." 5(6):670-671 [1974]);
cyclic carbonates such as (5-Rd-2-oxo-1,3-dioxolen-4-yl) methyl esters
(Sakamoto et al., "Chem. Pharm. Bull." 32(6):2241-2248 [19841) where Rd is R1,
R4 or aryl; and
-CH2C(O)N ~ 0
The hydroxyl 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 R6a ester moieties
that for example can be bonded via oxygen to -C(0)0- and -P(O)(O-)2 groups.
Several R6c amidates also are shown, which are bound directly to -C(O)- or
-P(0)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
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 W1 is 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 such as 13, 14 and 15)
with the monochlorophosphonate or dichlorophosphonate (or another
activated phosphonate).
TABLE A
1. -CH2-C(O)-N(R1)2 10. -CH2-O-C(O)-C(CH3)3
2. -CHZ-S(O)(Rl) 11. -CH2-CCl3
3. -CH2-S(0)2(R1) 12. -C6H5
4. -CH2-0-C(O)-CH2-C6-I5 13. -NH-CH2-C(O)O-CH2CH3
5. 3-cholesteryl 14. -N(CH3)-CH2-C(O)O-CH2CH3
6. 3-pyridyl 15. -NHR1
7. N-ethylmorpholino 16. -CH2-O-C(O)-C1oH15
8. -CH2-0-C(O)-C6H5 17. -CH2-0-C(O)-CH(CH3)2
9. -CH2-0-C(O)-CH2CH3 18. -CH2-C#H(OC(O)CH2R1)-CH2-
-(OC(O)CH2R1)
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WO 98/17647 PCTIUS97/18193
J\~ - , HO
2C(O)N O N\ ~ OH HO
-CH
19. 20. O H 21. HO
CH3O(O)C
N N
-CH2 O-C(O) -CH2CH2
22. 23. 24.
OCH3
CH3CH2O(O) -CH2 OCH3
~ ~ -
25. - 26. OCH3
# - chiral center is (R), (S) or racemate.
Other esters that are suitable for use herein are described in EP 632,048.
R6a also includes "double ester" forming profunctionalities such as
0
-CH2OC(O)OCH3, 0 ,-CH2SCOCH3, -CH2OCON(CH3)2, or alkyl- or
aryl-acyloxyalkyl groups of the structure -CH(R1 or W5)O((CO)R37) or -CH(Ri
or W5)((CO)OR38) (linked to oxygen of the acidic group) wherein R37 and R38
are alkyl, aryl, or alkylaryl groups (see U.S. Patent 4,968,788). Frequently
R37
and R38 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 pivaloyloxymethyl
group. These are of particular use with prodrugs for oral administration.
Examples of such useful R6a groups are alkylacyloxymethyl esters and their
derivatives, including -CH(CH2CH2OCH3)OC(O)C(CH3)3,
-,,~,O
TQ
; -CH2OC(O)C10Hj5, -CH2OC(O)C(CH3)3,
-CH(CH2OCH3)OC(O)C(CH3)3, -CH(CH(CH3)2)OC(O)C(CH3)3,
-CH2OC(O)CH2CH(CH3)2, -CH2OC(O)C6H11, -CH2OC(O)C6H5,
-CH2OC(O)C1oH15, -CH2OC(O)CH2CH3, -CH2OC(O)CH(CH3)2 ,
-CH2OC(O)C(CH3)3 and -CH2OC(O)CH2C6H5.
For prodrug purposes, the ester typically chosen is one heretofore used
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WO 98/17647 PCTIUS97/18193
for antibiotic drugs, in particular the cyclic carbonates, double esters, or
the
phthalidyl, aryl or alkyl esters.
As noted, R6a, R6c and R6b groups optionally are used to prevent side
reactions with the protected group during synthetic procedures, so they
function as protecting groups (PRT) during synthesis. For the most part the
decision as to which groups to protect, when to do so, and the nature of the
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 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 carboxyl, hydroxyl or amino groups.
The order of deprotection to yield free groups is dependent upon the intended
direction of the synthesis and the reaction conditions to be encountered, and
may occur in any order as determined by the artisan.
A very large number of R6a hydroxy protecting groups and R6c amide-
forming groups and corresponding chemical cleavage reactions are described
in "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 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 R6a carboxylic acid,
phosphonic acid, phosphonate, sulfonic acid and other protecting groups for
Wl acids see Greene as set forth below. Such groups include by way of
example and not limitation, esters, amides, hydrazides, and the like.
In some embodiments the R6a protected acidic group is an ester of the
acidic group and R6a is the residue of a hydroxyl-containing functionality. In
other embodiments, an R6c 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 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
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CA 02268756 2006-03-21
groups set forth herein and certain amino acid residues. Such residues are for
example set.
Typical R6a esters for protecting W1 acidic functionalities are also
described in 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 R31 or R35), the table ori 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-, alkoxy-, carboxy- and/or alkylestercarboxy-
substituted aryl or alkylaryl, especially phenyl, ortho-ethoxyphenyl, or Cl-C4
alkylestercarboxyphenyl (salicylate Cl-C12 alkylesters).
The protected acidic groups Wl, are useful as prodrugs for oral
administration. However, it is not essential that the Wl 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 free acid.
One or more of the acidic hydroxyls are protected. If more than one
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 R6a hydroxy protecting groups described in Greene j(pages 14-118)
include Ethers (Methyl); Substituted Methyl Ethers (Methoxymethyl,
Methylthiomethyl, t-Butylthiomethyl, (Pheriyldimethylsilyl)methoxymethyl,
Benzyloxymethyl, p-Methoxybenzyloxymethyl, (4-Methoxyphenoxy)methyl,
Guaiacolmethyl, t-Butoxymethyl, 4-Pentenyloxymethyl, Siloxymethyl,
2-Methoxvethoxymethyl, 2,2,2-Trichloroethoxymethyl, Bis(2-
chloroethoxy)methyl, 2-(Trimethylsilyl)ethoxymethyl, Tetrahydropyranyl,
3-Bromotetrahydropyranyl, Tetrahydropthiopyranyl, 1-Methoxycyclohexyl,
4-Methoxytetrahydropyranyl, 4-Methoxytetrahydrothiopyranyl,
4-Methoxytetrahydropthiopyranyl S,S-Dioxido, 1-1(2-Chloro-4-methyl)phenyl]-
4-methoxypiperidin-4-yl, 35, 1,4-Dioxan-2-yl, Tetrahydrofuranyl,
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CA 02268756 2006-03-21
Tetrahydrothiofuranyl, 2,3,3a,4,5,6,7,7a-Octahydro-7,8,8-trimethyl-4,7-
methanobenzofuran-2-yl)); Substituted Ethyl Ethers (1-Ethoxyethyl,
7-(2-Chloroethoxy)ethyl, 1-Methyl-l-methoxyethyl, 1-Methyl-l-benzyloxyethyl,
Methyl-l-benzyloxy-2-fluoroethyl, 2,2,2-Trichloroethyl, 2-
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WO 98/17647 PCT/US97/18193
Trimethylsilylethyl, 2- (Phenylselenyl) ethyl, t-Butyl, A11y1, p-Chlorophenyl,
p-
Methoxyphenyl,
2,4-Dinitrophenyl, Benzyl); Substituted Benzyl Ethers (p-Methoxybenzyl,
3,4-Dimethoxybenzyl, o-Nitrobenzyl, p-Nitrobenzyl, p-Halobenzyl,
2,6-Dichlorobenzyl, p-Cyanobenzyl, p-Phenylbenzyl, 2- and 4-Picolyl, 3-Methyl-
2-picolyl N-Oxido, Diphenylmethyl, p,p'-Dinitrobenzhydryl, 5-Dibenzosuberyl,
Triphenylmethyl, a-Naphthyldiphenylmethyl, p-
methoxyphenyldiphenylmethyl, Di(p-methoxyphenyl)phenylmethyl, Tri(p-
methoxyphenyl)methyl, 4-(4'-Bromophenacyloxy)phenyldiphenylmethyl,
4,4',4"-Tris(4,5-dichlorophthalimidophenyl)methyl, 4,4',4"-
Tris(levulinoyloxyphenyl)methyl, 4,4',4"-Tris(benzoyloxyphenyl)methyl,
3-(Imidazol-1-ylmethyl)bis(4',4"-dimethoxyphenyl)methyl, 1,1-Bis(4-
methoxyphenyl)-1'-pyrenylmethyl, 9-Anthryl, 9-(9-Phenyl)xanthenyl,
9-(9-Phenyl-10-oxo)anthryl, 1,3-Benzodithiolan-2-yl, Benzisothiazolyl S,S-
Dioxido); Silyl Ethers (Trimethylsilyl, Triethylsilyl, Triisopropylsilyl,
Dimethylisopropylsilyl, Diethylisopropylsily, Dimethylthexylsilyl, t-
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-Phenylpropionate, 4-Oxopentanoate (Levulinate), 4,4-
(Ethylenedithio)pentanoate, Pivaloate, Adamantoate, Crotonate,
4-Methoxycrotonate, Benzoate, p-Phenylbenzoate, 2,4,6-Trimethylbenzoate
(Mesitoate)); Carbonates (Methyl, 9-Fluorenylmethyl, Ethyl, 2,2,2-
Trichloroethyl, 2-(Trimethylsilyl)ethyl, 2-(Phenylsulfonyl)ethyl,
2-(Triphenylphosphonio)ethyl, Isobutyl, Vinyl, Allyl, p-Nitrophenyl, Benzyl,
p-Methoxybenzyl, 3,4-Dimethoxybenzyl, o-Nitrobenzyl, p-Nitrobenzyl, S-
Benzyl Thiocarbonate, 4-Ethoxy-l-naphthyl, Methyl Dithiocarbonate); Groups
With Assisted Cleavage (2-Iodobenzoate, 4-Azidobutyrate, 4-Niotro-4-
methylpentanoate, o-(Dibromomethyl)benzoate, 2-Formylbenzenesulfonate,
2-(Methylthiomethoxy)ethyl Carbonate, 4-(Methylthiomethoxy)butyrate,
2-(Methylthiomethoxymethyl)benzoate); Miscellaneous Esters (2,6-Dichloro-4-
methylphenoxyacetate, 2,6-Dichloro-4-(1,1,3,3
tetramethylbutyl)phenoxyacetate, 2,4-Bis(1,1-dimethylpropyl)phenoxyacetate,
Chorodiphenylacetate, Isobutyrate, Monosuccinoate, (E)-2-Methyl-2-butenoate
(Tigloate), o-(Methoxycarbonyl)benzoate, p-poly-Benzoate, a-Naphthoate,
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Nitrate, Alkyl N,N,N',N'-Tetramethylphosphorodiamidate, N-
Phenylcarbamate, Borate, Dimethylphosphinothioyl, 2,4-
Dinitrophenylsulfenate); and Sulfonates (Sulfate, Methanesulfonate
(Mesylate), Benzylsulfonate, Tosylate).
More typically, R6a hydroxy protecting groups include substituted
methyl ethers, substituted benzyl ethers, silyl ethers, and esters including
sulfonic acid esters, still more typically, trialkylsilyl ethers, tosylates
and
acetates.
Typical 1,2-diol protecting groups (thus, generally where two OH groups
are taken together with the R6a protecting functionality) are described in
Greene at 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 Ortho Esters
(Methoxymethylene, Ethoxymethylene, Dimethoxymethylene,
1-Methoxyethylidene, 1-Ethoxyethylidine, 1,2-Dimethoxyethylidene,
a-Methoxyberizylidene, 1-(N,N-Dimethylamino)ethylidene Derivative,
(x-(N,N-Dimethylamino)benzylidene Derivative, 2-Oxacyclopentylidene);
Silyl Derivatives (Di-t-butylsilylene Group, 1,3-(1,1,3,3-
Tetraisopropyldisiloxanylidene), and Tetra-t-butoxydisiloxane-1,3-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.
Table B
~~, r~c r~c r~c r~c 0
O o
O O o o
O \ S ,o o\
y
11
O o 0 SO
r~c r~c r~c r~c r
O"O ,N O 9 9 O
R90 ~ R O-N, /O R O-N O
R90 O O O S~~O R90 P\\O
0
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WO 98/17647 PCT/US97/18193
wherein R9 is C1-C6 alkyl.
R6b is H, a protecting group for amino or the residue of a carboxyl-
containing compound, in particular H, -C(O)R4, an amino acid, a polypeptide
or a protecting group not -C(O)R4, amino acid or polypeptide. Amide-forming
R(b are found for instance in group G1. When R6b is an amino acid or
polypeptide it has the structure R15NHCH(R16)C(O)-, where R15 is H, an
amino acid or polypeptide residue, or R5, and R16 is defined below.
R16 is lower alkyl or lower alkyl (C1-C6) substituted with amino,
carboxyl, amide, carboxyl ester, hydroxyl, C6-C7 aryl, guanidinyl, imidazolyl,
indolyl, sulfhydryl, sulfoxide, and/or alkylphosphate. R16 also is taken
together with the amino acid (x N to form a proline residue (R16 = -CH2)3-).
However, R16 is generally the side group of a naturally-occurring amino acid
such as H, -CH3, -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. R16 also includes 1-guanidinoprop-3-yl, benzyl, 4-
hydroxybenzyl, imidazol-4-yl, indol-3-yl, methoxyphenyl and ethoxyphenyl.
R6b are residues of carboxylic acids for the most part, but any of the
typical amino protecting groups described by Greene at pages 315-385 are
useful. They include Carbamates (methyl and ethyl, 9-fluorenylmethyl, 9(2-
sulfo)fluoroenylmethyl, 9-(2,7-dibromo)fluorenylmethyl, 2,7-di-t-buthyl-[9-
(10,10-dioxo-10,10,10,10-tetrahydrothioxanthyl)]methyl, 4-methoxyphenacyl);
Substituted Ethyl (2,2,2-trichoroethyl, 2-trimethylsilylethyl, 2-phenylethyl,
1-(1-adamantyl)-1-methylethyl, 1,1-dimethyl-2-haloethyl, 1,1-dimethyl-2,2-
dibromoethyl, 1,1-dimethyl-2,2,2-trichloroethyl, 1-methyl-l-(4-
biphenylyl)ethyl, 1-(3,5-di-t-butylphenyl)-1-methylethyl, 2-(2'- and 4'-
pyridyl)ethyl, 2-(N,N-dicyclohexylcarboxamido)ethyl, t-butyl, 1-adamantyl,
vinyl, allyl,
1-isopropylallyl, cinnamyl, 4-nitrocinnamyl, 8-quinolyl, N-hydroxypiperidinyl,
alkyldithio, benzyl, p-methoxybenzyl, p-nitrobenzyl, p-bromobenzyl, p-
chorobenzyl, 2,4-dichlorobenzyl, 4-methylsulfinylbenzyl, 9-anthrylmethyl,
diphenylmethyl); Groups With Assisted Cleavage (2-methylthioethyl,
2-methylsulfonylethyl, 2-(p-toluenesulfonyl)ethyl, [2-(1,3-dithianyl)]methyl,
4-methylthiophenyl, 2,4-dimethylthiophenyl, 2-phosphonioethyl,
2-triphenylphosphonioisopropyl, 1,1-dimethyl-2-cyanoethyl, m-choro-p-
acyloxybenzyl, p-(dihydroxyboryl)benzyl, 5-benzisoxazolylmethyl,
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2-(trifluoromethyl)-6-chromonylmethyl); Groups Capable of Photolytic
Cleavage (m-nitrophenyl, 3,5-dimethoxybenzyl, o-nitrobenzyl, 3,4-dimethoxy-
6-nitrobenzyl, phenyl(o-nitrophenyl)methyl); Urea-Type Derivatives
(phenothiazinyl-(10)-carbonyl, N'-p-toluenesulfonylaminocarbonyl, N'-
phenylaminothiocarbonyl); Miscellaneous Carbamates (t-amyl, S-benzyl
thiocarbamate, p-cyanobenzyl, cyclobutyl, cyclohexyl, cyclopentyl,
cyclopropylmethyl, p-decyloxybenzyl, diisopropylmethyl, 2,2-
dimethoxycarbonylvinyl, o-(N,N-dimethylcarboxamido)benzyl, 1,1-dimethyl-
3-(N,N-dimethylcarboxamido)propyl, 1,1-dimethylpropynyl, di(2-
pyridyl)methyl, 2-furanylmethyl, 2-lodoethyl, Isobornyl, Isobutyl,
Isonicotinyl,
p-(p'-Methoxyphenylazo)benzyl, 1-methylcyclobutyl, 1-methylcyclohexyl, 1-
methyl-l-cyclopropylmethyl, 1-methyl-l-(3,5-dimethoxyphenyl)ethyl, 1-
methyl-l-(p-phenylazophenyl)ethyl, 1-methyl-l-phenylethyl, 1-methyl-l-(4-
pyridyl)ethyl, phenyl, p-(phenylazo)benzyl, 2,4,6-tri-t-butylphenyl, 4-
(trimethylammonium)benzyl, 2,4,6-trimethylbenzyl); Amides (N-formyl, N-
acetyl, N-choroacetyl, N-trichoroacetyl, N-trifluoroacetyl, N-phenylacetyl, N-
3-
phenylpropionyl, N-picolinoyl, N-3-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-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-nitrobenzoyl, N-
o-(benzoyloxymethyl)benzoyl, 4,5-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 1,3-dimethyl-1,3,5-triazacyclohexan-2-one, 5-substituted 1,3-
dibenzyl-1,3-5-triazacyclohexan-2-one, 1-substituted 3,5-dinitro-4-pyridonyl);
N-Alkyl and N-Aryl Amines (N-methyl, N-allyl, N-[2-
(trimethylsilyl)ethoxyjmethyl, N-3-acetoxypropyl, N-(1-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-
methoxyphenyl)diphenylmethyl, N-9-phenylfluorenyl, N-2,7-dichloro-9-
fluorenylmethylene, N-ferrocenylmethyl, N-2-picolylamine N'-oxide), Imine
Derivatives (N-1,1-dimethylthiomethylene, N-benzylidene, N-p-
methoxybenylidene, N-diphenylmethylene, N-[(2-pyridyl)mesityl]methylene,
N,(N',N'-dimethylaminomethylene, N,N'-isopropylidene, N-p-
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nitrobenzylidene, N-salicylidene, N-5-chlorosalicylidene, N-(5-chloro-2-
hydroxyphenyl)phenylmethylene, N-cyclohexylidene); Enamine Derivatives
(N-(5,5-dimethyl-3-oxo-l-cyclohexenyl)); N-Metal Derivatives (N-borane
derivatives, N-diphenylborinic acid derivatives, N-
[phenyl(pentacarbonylchromium- or -tungsten)]carbenyl, N-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); N-Si
Derivatives; N-S Derivatives; N-Sulfenyl Derivatives (N-benzenesulfenyl, N-
o-nitrobenzenesulfenyl, N-2,4-dinitrobenzenesulfenyl, N-
pentachlorobenzenesulfenyl, N-2-nitro-4-methoxybenzenesulfenyl, N-
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-methoxybenzenesulfonyl,
N-2,4,6-trimethylbenzenesulfonyl, N-2,6-dimethoxy-4-
methylbenzenesulfonyl, N-2,2,5,7,8-pentamethylchroman-6-sulfonyl, N-
methanesulfonyl, N-[3-trimethylsilyethanesulfonyl, N-9-anthracenesulfonyl,
N-4-(4',8'-dimethoxynaphthylmethyl)benzenesulfonyl, N-benzylsulfonyl, N-
trifluoromethylsulfonyl, N-phenacylsulfonyl).
More typically, protected amino groups include carbamates and amides,
still more typically, -NHC(O)R1 or -N=CR1N(Rl)2. Another protecting group,
also useful as a prodrug at the G1 site, particularly for amino or -NH(R5),
is:
O
O"k O 0
- ~---~
Ws ~ O
see for example Alexander, J. et al., "J. Med. Chem." 39:480-486 (1996).
R6c is H or the residue of an amino-containing compound, in particular
an amino acid, a polypeptide, a protecting group, -NHSO2R4, NHC(O)R4,
-N(R4)2, NH2 or -NH(R4)(H), whereby for example the carboxyl or phosphonic
acid groups of Wi are reacted with the amine to form an amide, as in -C(O)R6c,
-P(O)(R6c)2 or -P(O)(OH)(R6c). In general, R6c has the structure
R17C(O)CH(R16)NH-, where R17 is OH, OR6a, OR5, an amino acid or a
polypeptide residue.
Amino acids are low molecular weight compounds, on the order of less
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than about 1,000 MW, that contain at least one amino or imino group and at
least one carboxyl group. Generally 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,
i.e. 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 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
naturally in plants, animals or microbes, especially proteins thereof.
Polypeptides most typically will be substantially composed of such naturally-
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.
When R6b and R6c are single amino acid residues or polypeptides they
usually are substituted at R3, W6, W1 and/or W2, but typically only Wi or
W2. These conjugates are produced by forming an amide bond between a
carboxyl group of the amino acid (or C-terminal amino acid of a polypeptide
for example) and W2. Similarly, conjugates are formed between W1 and an
amino group of an amino acid or polypeptide. Generally, only one of any site
in the parental molecule 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 Wi is amidated
with an amino acid. In general, the a-amino or a-carboxyl group of the amino
acid or the terminal amino or carboxyl group of a polypeptide are bonded to
the parental functionalities, i.e., carboxyl or amino groups in the amino acid
side chains generally are not used to form the amide bonds with the parental
compound (although 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
polypeptides it will be understood that the carboxyl group optionally will be
blocked, e.g. by R6a, esterified with R5 or amidated with R6c. Similarly, the
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amino side chains R16 optionally will be blocked with R6b or substituted with
R5.
Such ester or amide bonds with side chain amino or carboxyl groups,
like the esters or amides with the parental molecule, optionally are
hydrolyzable in vivo or in vitro under 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 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 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 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 R6b
and R6c include the following:
Glycine;
Aminopolycarboxylic acids, e.g., aspartic acid, 0-hydroxyaspartic acid,
glutamic acid, (3-hydroxyglutamic acid, P-methylaspartic acid, 0-
methylglutamic acid, (3,(3-dimethylaspartic acid, y-hydroxyglutamic acid, 0,7
dihydroxyglutamic acid, (3-phenylglutamic acid, y-methyleneglutamic acid, 3-
aminoadipic acid, 2-aminopimelic acid, 2-aminosuberic acid and 2-
aminosebacic acid;
Amino acid amides such as glutamine and asparagine;
Polyamino- or polybasic-monocarboxylic acids such as arginine, lysine,
0-aminoalanine, y-aminobutyrine, ornithine, citruline, homoarginine,
homocitrulline, hydroxylysine, allohydroxylsine and diaminobutyric acid;
Other basic amino acid residues such as histidine;
Diaminodicarboxylic acids such as a,a'-diaminosuccinic acid, a,a'-
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diaminoglutaric acid, a,a'-diaminoadipic acid, a,a'-diaminopimelic acid, a,a'-
diamino-(3-hydroxypimelic acid, a,a'-diaminosuberic acid, a,a'-
diaminoazelaic acid, and a,a'-diaminosebacic acid;
Imino acids such as proline, hydroxyproline, allohydroxyproline, y-
methylproline, pipecolic acid, 5-hydroxypipecolic acid, and azetidine-2-
carboxylic acid;
A mono- or di-alkyl (typically C1 - C8 branched or normal) amino acid
such as alanine, valine, leucine, allylglycine, butyrine, norvaline,
norleucine,
heptyline, a-methylserine, a-amino-a-methyl-y-hydroxyvaleric acid, a-amino-
a-methyl-b-hydroxyvaleric acid, a-amino-a-rnethyl-~-hydroxycaproic acid,
isovaline, a-methylglutamic acid, a-aminoisobutyric acid, a-
aminodiethylacetic acid, a-aminodiisopropylacetic acid, a-aminodi-n-
propylacetic acid, a-aminodiisobutylacetic acid, a-aminodi-n-butylacetic acid,
a-aminoethylisopropylacetic acid, a-amino-n-propylacetic acid, a-
aminodiisoamyacetic acid, a-methylaspartic acid, a-methylglutamic acid, 1-
arninocyclopropane-l-carboxylic acid, isoleucine, alloisoleucine, tert-
leucine,
j3-methyltryptophan and a-amino-(3-ethyl-(3-phenylpropionic acid;
0-phenylserinyl;
Aliphatic a-amino-(3-hydroxy acids such as serine, (3-hydroxyleucine, 0-
hydroxynorleucine, 0-hydroxynorvaline, and a-amino-(3-hydroxystearic acid;
a-Amino, a-, y-, S- or e-hydroxy acids such as homoserine, y-
hydroxynorvaline, S-hydroxynorvaline and epsilon-hydroxynorleucine
residues; canavine and canaline; y-hydroxyornithine;
2-hexosaminic acids such as D-glucosaminic acid or D-galactosaminic
acid;
a-Amino-(3-thiols such as penicillamine, 0-thiolnorvaline or P-
thiolbutyrine;
Other sulfur containing amino acid residues including cysteine;
hornocystine, P-phenylmethionine, methionine, S-allyl-L-cysteine sulfoxide,
2-thiolhistidine, cystathionine, and thiol ethers of cysteine or homocysteine;
Phenylalanine, tryptophan and ring-substituted a amino acids such as
the phenyl- or cyclohexylamino acids a-aminophenylacetic acid, a-
aminocyclohexylacetic acid and a-amino-p-cyclohexylpropionic acid;
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-dicloro, o-, m-
or p-
methyl-, 2,4,6-trimethyl-, 2-ethoxy-5-nitro-, 2-hydroxy-5-nitro- and p-nitro-
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phenylalanine); furyl-, thienyl-, pyridyl-, pyrimidinyl-, purinyl- or naphthyl-
alanines; and tryptophan analogues and derivatives including kynurenine, 3-
hydroxykynurenine, 2-hydroxytryptophan and 4-carboxytryptophan;
a-Amino substituted amino acids including sarcosine (N-
methylglycine), N-benzylglycine, N-methylalanine, N-benzylalanine, N-
methylphenylalanine, N-benzylphenylalanine, N-methylvaline and N-
benzylvaline; and
a-Hydroxy and substituted a-hydroxy amino acids including serine,
threonine, allothreonine, phosphoserine and phosphothreonine.
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
molecular weight polypeptides (about 1500-5000MW) 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
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 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 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 not need to be immunogenic in
an animal typically used to raise antibodies, e.g., rabbit, mouse, horse, or
rat,
but the final product conjugate should be immunogenic in at least one of such
animals. 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.
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Peptidolytic enzymes for cleaving the polypeptide conjugates of this
invention are well known, and in particular include carboxypeptidases.
Carboxypeptidases 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 given pair of residues and a free carboxyl
terminus) is covalently bonded through its a-amino group to the phosphorus
or carbon atoms of the compounds herein. In embodiments where W1 is
phosphonate it is expected that this 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, Al, AL, AK, AM, AF, AP, AS, AT,
AW, AY, 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, EK, EM, EF, EP, ES, ET, EW, EY, EV, QA, QR, QN, QD, QC,
QE, QQ, 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, HC, HE, HQ, HG, HH, HI, HL, HK, HM, HF, HP, HS, HT, HW,
HY, HV, IA, IR, IN, ID, IC, IE, IQ, IG, IH, II, IL, IK, IM, IF, IP, IS, IT,
IW, IY, IV,
LA, LR, LN, LD, LC, LE, LQ, LG, LH, LI, LL, LK, LM, LF, LP, LS, LT, LW, LY,
LV,
KA, KR, KN, KD, KC, KE, KQ, KG, KH, KI, KL, KK, KM, KF, KP, KS, KT, KW,
KY, KV, MA, MR, MN, MD, MC, ME, MQ, MG, MH, MI, ML, MK, MM, MF,
MP, MS, MT, MW, MY, MV, FA, FR, FN, FD, FC, FE, FQ, FG, FH, Fl, FL, FK,
FM, FF, FP, FS, FT, FW, FY, FV, PA, PR, PN, PD, PC, PE, PQ, PG, PH, PI, PL,
PK,
PM, PF, PP, PS, PT, PW, PY, PV, SA, SR, SN, SD, SC, SE, SQ, SG, SH, SI, SL,
SK,
SM, SF, SP, SS, ST, SW, SY, SV, TA, TR, TN, TD, TC, TE, TQ, 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, YQ, YG, YH, YI, YL, YK, YM, YF, YP, YS, YT, YW, YY, YV, VA,
VR, VN, VD, VC, VE, VQ, VG, VH, VI, VL, VK, VM, VF, VP, VS, VT, VW,
VY and VV.
Tripeptide residues are also useful as R6b or R6c. When Wi is
phosphonate, the sequence -X4-pro-X5- (where X4 is any amino acid residue
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and X5 is an amino acid residue, a carboxyl ester of proline, or hydrogen)
will
be cleaved by luminal carboxypeptidase to yield X4 with a free carboxyl, which
in turn is expected to autocatalytically cleave the phosphonoamidate bond.
The carboxy group of X5 optionally is esterified with benzyl.
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
a-amino group are transport substrates for the peptide transporter found in
brush border membrane of intestinal mucosal cells (Bai, J.P.F., "Pharm Res."
9:969-978 (1992). 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 are also compatible with peptide
transport and can be utilized in the amidate compounds of this invention.
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 aminopeptidase N (EC 3.4.11.2). 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 (EC 3.4.11.7),
di- or tripeptides lacking amino acid residues on the N-terminal side of
hydrophobic amino acids (leu, tyr, phe, val, trp) are poor substrates for
endopeptidase 24.11 (EC 3.4.24.11), and peptides lacking a pro residue at the
penultimate position at a free carboxyl terminus are poor substrates for
carboxypeptidase P (EC 3.4.17). 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.
Such poorly cleaved polypeptide amidates are immunogens or are useful for
bonding to proteins in order to prepare immunogens.
Stereoisomers
The compounds of the invention are enriched or resolved optical
isomers at any or all asymmetric atoms. For example, the chiral centers
apparent from 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.
One or more of the following enumerated methods are used to prepare
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the enantiomerically enriched or pure isomers herein. The methods are listed
in approximately their order of preference, i.e., one ordinarily should employ
stereospecific synthesis from chiral precursors before chromatographic
resolution before spontaneous crystallization.
Stereospecific synthesis is described below. Methods of this type
conveniently are used when the appropriate chiral starting material is
available and reaction steps are chosen do not result in undesired
racemization at chiral sites. One advantage of stereospecific synthesis is
that it
does not produce undesired enantiomers that must be removed from the final
product, thereby lowering overall synthetic yield. In general, those skilled
in
the art would understand what starting materials and reaction conditions
should be used to obtain the desired enantiomerically enriched or pure
isomers by stereospecific synthesis. If an unexpected racemization occurs in a
method thought to be stereospecific then one needs only to use one of the
following separation methods to obtain the desired product.
If a suitable stereospecific synthesis cannot be empirically designed or
determined with routine experimentation then those skilled in the art would
turn to other methods. One method of general utility is chromotographic
resolution of enantiomers on chiral chromatography resins. These resins are
packed in columns, commonly called Pirkle columns, and are commercially
available. The columns contain a chiral stationary phase. The racemate is
placed in solution and loaded onto the column, and thereafter separated by
HPLC. See for example, Proceedings Chromatographic Society - International
Symposium on Chiral Separations, Sept. 3-4, 1987. Examples of chiral
columns that could be used to screen for the optimal separation technique
would include Diacel Chriacel OD, Regis Pirkle Covalent Dphenylglycine,
Regis Pirkle Type 1A, Astec Cyclobond II, Astec Cyclobond III, Serva Chiral D-
DL=Daltosil 100, Bakerbond DNBLeu, Sumipax OA-1000, Merck Cellulose
Triacetate column, Astec Cyclobond I-Beta, or Regis Pirkle Covalent D-
Naphthylalanine. Not all of these columns are likely to be effective with
every racemic mixture. However, those skilled in the art understand that a
certain amount of routine screening may be required to identify the most
effective stationary phase. When using such columns it is desirable to employ
embodiments of the compounds of this invention in which the charges are
not neutralized, e.g., where acidic functionalities such as carboxyl are not
esterified or amidated.
Another method entails converting the enantiomers in the mixture to
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diastereomers with chiral auxiliaries and then separating the conjugates by
ordinary column chromatography. This is a very suitable method, particularly
when the embodiment contains free carboxyl, amino or hydroxyl that will
form a salt or covalent bond to a chiral auxiliary. Chirally pure amino acids,
organic acids or organosulfonic acids are all worthwhile exploring as chiral
auxiliaries, all of which are well known in the art. Salts with such
auxiliaries
can be formed, or they can be covalently (but reversibly) bonded to the
functional group. For example, pure D or L amino acids can be used to
amidate the carboxyl group of embodiments of this invention and then
separated by chromatography.
Enzymatic resolution is another method of potential value. In such
methods one prepares covalent derivatives of the enantiomers in the racemic
mixture, generally lower alkyl esters (for example of carboxyl), and then
exposes the derivative to enzymatic cleavage, generally hydrolysis. For this
method to be successful an enzyme must be chosen that is capable of
stereospecific cleavage, so it is frequently necessary to routinely screen
several
enzymes. If esters are to be cleaved, then one selects a group of esterases,
phosphatases, and lipases and determines their activity on the derivative.
Typical esterases are from liver, pancreas or other animal organs, and include
porcine liver esterase.
If the enantiomeric mixture separates from solution or a melt as a
conglomerate, i.e., a mixture of enantiomerically-pure crystals, then the
crystals can be mechanically separated, thereby producing the enantiomerically
enriched preparation. This method, however, is not practical for large scale
preparations and is of no value for true racemic compounds.
Asymmetric synthesis is another technique for achieving enantiomeric
enrichment. For example, a chiral protecting group is reacted with the group
to be protected and the reaction mixture allowed to equilibrate. If the
reaction
is enantiomerically specific then the product will be enriched in that
enantiomer.
Further guidance in the separation of enantiomeric mixtures can be
found, by way of example and not limitation, in "Enantiomers, Racemates,
and resolutions", Jean Jacques, Andre Collet, and Samuel H. Wilen (Krieger
Publishing Company, Malabar, FL, 1991, ISBN 0-89464-618-4). In particular,
Part 2, Resolution of Enantiomer Mixture, pages 217-435; more particularly,
section 4, Resolution by Direct Crystallization, pages 217-251, section 5,
Formation and Separation of Diastereomers, pages 251-369, section 6,
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Crystallization-Induced Asymmetric Transformations, pages 369-378, and
section 7, Experimental Aspects and Art of Resolutions, pages 378-435; still
more particularly, section 5.1.4, Resolution of Alcohols, Transformation of
Alcohols into Salt-Forming Derivatives, pages 263-266, section 5.2.3, Covalent
Derivatives of Alcohols, Thiols, and Phenols, pages 332-335, section 5.1.1,
Resolution of Acids, pages 257-259, section 5.1.2, Resolution of Bases, pages
259-260, section 5.1.3, Resolution of Amino Acids, page 261-263, section
5.2.1,
Covalent Derivatives of Acids, page 329, section 5.2.2, Covalent derivatives
of
Amines, pages 330-331, section 5.2.4, Covalent Derivatives of Aldehydes,
Ketones, and Sulfoxides, pages 335-339, and section 5.2.7, Chromatographic
Behavior of Covalent Diastereomers, pages 348-354, are cited as examples of
the skill of the art.
The compounds of the invention can also exist as tautomeric isomers
in certain cases. For example, ene-amine tautomers can exist for imidazole,
guanidine, amidine, and tetrazole systems and all their possible tautomeric
forms are within the scope of the invention.
Exemplary Enumerated Compounds.
By way of example and not limitation, embodiment compounds are
named below in tabular format (Table 6). Generally, each compound is
depicted as a substituted nucleus in which the nucleus is designated by
capital
letter and each substituent is designated in order by lower case letter or
number. Table 1 are a schedule of nuclei which differ principally by the
position of ring unsaturation and the nature of ring substituents. Each
nucleus is given a alphabetical designation from Table 1, and this designation
appears first in each compound name. Similarly, Tables 2, 3, 4, and 5 list the
selected Ql, Q2, Q3 and Q4 substituents, again by letter or number
designation.
Accordingly, each named compound will be depicted by a capital letter
designating the nucleus from Table 1, followed by a number designating the
Q1 substituent, a lower case letter designating the Q2 substituent, a number
designating the Q3 substituent, and a lower case letter or letters designating
the
Q4 substituent. Thus, the structure below has the name shown.
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H3C H3C
N CO2CH2CH3
H\N I
H3CO NH2
A.141.x.4. i
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Table 1
H
I
Q1-~ Q2 Q111 N/i.. \ Q2 Q1-Q/i, Q2
Q4 Q4 N Q4 N
Qg Q3 Q3
A B C
H
Q1--,N Q2 Q1NqQ2 Q1,Q/,, Q2
Q4 Q4 Q4
Qg Q3 Q3
D E F
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Table 2a
H-Q1 H3C-Q1 H3C\/Q1 H3C,,/-, Qy
1 2 3 4
H3C Q1 ~
~ Q1 H3C~\Q1 H3C
H3C H3C " ' Q1
6 7 8
H3C ~Q1 H3C Q1 O
Q~ y H3C-"A
H3C O Q1
9 10 11 12
0 O
H3C Q H3C Q1 ~~iQi HO~Q~
-)A O
H3C
13 14 15 16
HO~~Qi HO~-'Qy H3C I Qi HO--~Q1
OH H3C
17 18 19 20
H3CQ1 OH HO H3C
OH H3C Qi H3C " 01 HO- Q1
21 22 23 24
0 0
HO ""~Ql HO -'~)rQ1 H3C * ~
H3C Q1 HO Q1
O
OH
25 26 27 28
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Table 2b
O
OH O O O
*
H3C Qi H3C ')* HO ~Q> HO~Q
~ ~ Q 1 >
OH H3C
29 30 31 32
H2N/-"iQ1 H2N/-~'~Qi H3C * Q~
Q1 H2N I
NH2 H3C
33 34 35 36
H3C/~ Q1 N1 H2 H2N H3C
NH2 H3C *1- ~~Q1 H3C * Q1 H2N~Q1
37 38 39 40
O
H2N~\Qt H2N-'~~y Q1 H3C * O
H C Q1
3 0 H2N--~Qi
NH2
41 42 43 44
O NH2 O O O O
~Qi H3C H3C kH2N~Q1 HO ~Q
1
NH2 H3C
45 46 47 48
HO~~Q~ HO Ql OH HO -'y OH H3C * * Q1 H3C
Q1
HO OH OH
49 50 51 52
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Table 2c
HO H3C O
HOQ1 HO * Q1
HO Q1 OH HO OH
53 54 55 56
0
OH O 0 OH O
* HO *
H3C Q1 Q 1 HO,,~ HO Q1
OH OH Ql HO
57 58 59 60
Q1 NH2 H2N
H2NQl H2N
NH2 H3C * * Q1 H3C
HN NH2 Q1
NH2
61 62 63 64
H2N H3C 0
H2N Ql H2N Ql H2N * Q1
H2N * Ql NH2 H2N NH2
65 66 67 68
NH2 O 0
* H2N * 0
H3C Q1 Qy 2 1 H2N 01
H N DI Q ~
NH2 NH2 H2N
69 70 71 72
01 OH
HOQ1 H2NQ1 HO H3C ** Qi
NH2 OH H2N NH2
73 74 75 76
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Table 2d
NH2 HO H2N H3C
H3C ** Q1 H3C H3C HO
Q1 Q Q1
OH l NH2
NH2 OH
77 78 79 80
H3C HO H2N
HO * 01
H2N * * Q1
OH H2N * Q1 HO Qi H2N
81 82 83 84
0 OH NH2 O
HO""~Q1 H2N J*Q, H3C Qi H C * Q
3 1
NH2 OH NH2 OH
85 86 87 88
O O O
HO Q1 H2N~Q1 H2NJI'~Q1 H2NJ"AQ1
0
H2N
89 90 91 92
H2N\ ~Q1 NH NH ~
1(
NH H2N~/\Q1 H2N~~Q1 H2N Q1
93 94 95 96
H3C.0 ~Q1 H3C' O"-,,-~Q1
97 98
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Table 2e
Qt--ir'CH3 Qt,~,~ CH3 QtCH3 Q1~/~
CH3 CH3 H3C/ CH3 CH3
100 101 102 103
Q1-,,--~CH3 Qt,,~~CH3 CH3 CH3
Qt~iC-13 Q1CH3
CH3 CH3
104 105 106 107
Q
Qt,,-/'~ CH3 t CH3 p CH3 H3C CH3
I CH3 t~CH3 Q"~XCH3 CH3 CH3
CH3
108 109 110 111
Qt CH3
~CH3 Qt ~ Qt CH3 Qt-'~~~CH3
H3C CH3 CH3 CH3
112 113 114 115
pt CH3 CH3 Qt'~/~
CH
CHs 3
CH3 Qt CH Qt~~CH3
CH3 3
116 117 118 119
CH3 Qt\ CH3 Q1~iCH3
pt CH3 Qt ~
CH3 CH3 CH3 CH3
120 121 122 123
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Table 2f
Q CH3 CH3 CH3 CH3
~ Q1\~/CH3 Q1 = CH3 Qilll~~CI.i3
H3C CH3
CH3 CH3 CH3
124 125 126 127
CH3 H C CH
Q1~~CH3 Q1~CH3 Q1~CH3 Q1 3 3
CH3 CH3 CH3 CH3 CH3
CH3 CH3
128 129 130
131
H3C~ ,CH3 CH3 CH3
Q~CH3 Q1\ ~CH3 QH1C CH3 Q1 CH3
CH3 CH
3 3 CH3
132 133 134 135
CH3 CH3 C H3
Q1 H3 Q1 CH3 Qi~\~ CH3
Q1 v CH3 H3~C
~ ~ I
CH3 CH3 CH3
136 137 138 139
Q1~CH3 CH3
Q1~CH3 Q1 CH3 Q1,~~'CH3
H3C CH3 CH3
140 141 142 143
Q1 CH3 CH3 CH3
Q1 CH3 Qj - CH3
CH3
144 145 146
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Table 2g
Q1-,,,-~CH3 Q1 - CH3 CH3
~ Q1 CH3
CH3 CH3
147 148 149
C_H3
Q1 CH3 Q CH3 Q1 CH3
CH3 C~H3
0 151 152
1 CH3 CH3
Q1~/~CH3 Q1---' \CH3 Q ~"~CH3
H3C CH3 ~~---~ CH3
Q
CH3 CH3 CH3
153 154 155 156
CH3 CH3
Q1~~CH3 Q1~~~CH3 Q1\~CH3 Q1\~CH3
CH3 CH3 CH3 CH3 CH3 CH3
157 158 159 160
Q1 Q1'-,/\~'~ CH3 C, H3
CH3 = = CH3 Q1\~CH3 Q1''~~v~'
CH3 CH3 CH3 CH3 CH3
CH3 CH3
161 162 163 164
CH3 ~CH3 H3C\ /CH3 CH3
Q1'~, Q1-/\~CH Q1'~~./~CH3 Ql~~~CH3
CHg 3 CH3
165 166 167 168
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Table 2h
CH3 CH3 CH3 CH3 CH3
Q~~CH3 Q1~'~ CH3 Q1w ~CH3 Q1~~CH
CH3 3
CH3 CH3
169 170 171 172
Q1 CH3 Q1 CH3 CH3
\~~ --'~CH3 Q1"~CH Q1~CH3
H3C CH3 3
CH3 CH3 CH3
173 174 175 176
Q1 CH3 Q1 CH3 Qj,,,~~CH3 Q1CH3
3
CH3 H3C~C-13 H3C ~ CH3 H 3C~ CH
177 178 179 180
CH3 CH3 C_H3
Ql~CH3 Q1 CH3 Q1~~CH3 Q1 CH3
H3C ' CH3 "CH3
CH3 CH3 181 182 183 184
C_H3
Q1,,,~CH3 Q1 CH3 Qi-,~CH3 H3C CH3
~ H3 CH3
CH3 CH Q1CH3
A-",c CH3 3
185 186 187 188
H3C~CH3 H C CH CH3 CH3
3 3
CH3 Q1 CHg
CH3 Q1 CH3 Q1
Q1~i-,~c
YC
CH3 CH3
189 190 191 192
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Table 2i
CH3 H3C CH3
Qt Q1 CH3 Q1~iCH3
.~
CH3 QlCH3
H3C CH3 H3C CH3 H3CCH3 CH3
193 194 195 196
Q1~~CH3 CH3 CH3 CH3
Q1~CH3 Qi~CH3 QCH3
H3C CH3CH3 7 T
CH3 CH3 CH3 CH3 CH3 CH3
197 198 199 200
CH3 Q1 Ql
Q1~/CH3 H3C 8H
CH3 CT H3 01 CH3 H3C 3 H3C
201 202 203 204
CH3 C_ H3 H3C H3C
Q1 Q Q1 Q1
H3C H3C CH3 CH3
205 206 207 208
H C H C H3C CFi3 H3C 11J=CH3
3 3
Q1 CHg Q1 CHg Q1 Q1
209 210 211 212
CH3 H3C H3C H3C
H C Q1 Q1~ Q1
3
Q1 H3C CH3
CH3 CH3
213 214 215 216
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Table 2j
CH3 CH3 CH3 CH3
Q~ Q1 Q1 Q1
CH3 CH3 CH3 CH3 CH3 CH3 CH3 CH3
217 218 219 220
H3C H3C H3C H3C
Q1 Q1 Q1 Q1"~/
CH3 CH3 CH3 CH3 CH3 CH3 CH3 CH3
221 222 223 224
H3C Ha C H3C H3C
Q1 Q1~~./ Q1 "/i Q1"' i
CH3 CH3 CH3 CH3
CH3 CH3 CH3 CH3
225 226 227 228
H3C CH3 H3C CH3 CH3 "CH3
Q~ Q1'~.,~ V Q1 Q1
CH3 CH3 CH3 CH3
229 230 231 232
H3C CH3 CH3 C H3 CH3
Q1 Qi Q1 Q1
CH3 CH3 CH3 CH3 CH3 CH3 CH3
233 234 235 236
C_H3 CH3 CH3 CH3 CH3 CH3 CH3
Q1 Q~ Q1 - (~1 '
CH3 CH3 CH3 CH3 CH3
237 238 239 240
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Table 2k
CH3 CH3 Q1 CH3 CHg H3C
Q1 _ Q1
Q~
CH3 H3C H3C
CH3 CH3 CH3
241 242 243 244
H3C H3C H3C H3C
Q1~ Q1 Q1 Q1~
CN3 CH3
CH3 H3C CH3 CH3 6H3
245 246 247 248
H3C N3C H3C CH3 H3C Q1 '=,i Q1.a" CHCH XCH3
3
CH3 CH3 CH3 CH3
249 250 251 252
Q1 Q1 Q1 Ql
N3C CH3 H31~'CH3 H3C H3C c1CH3
CH3 H3C CH3
253 254 255 256
Q'
CH3 Q1--~~CH3 Q.J1 ~j CH3 Ql~/~CH3
CH3 ~CH3 ~~ ~ ~
H3C CH3 H3C CH3
257 258 259 260
CH3 CH3 CH3 CH3
Q1~~~ CH3 Q1l~r~~CH3 Q1 Q~W
Nl-
N3C f CN3 H3C ' CH3
CH3 CH3
261 262 263 264
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Table 21
CH3 CH3 CH3 CH3 CH3 CH3
Q1 Q
Q1~
CH3 CH3 H3C CH3 H3Ci CH3
265 266 267 268
CH3 CH3 CH3 CH3
Q1 = Q1~ Q1 CH3 Q1~CH3
H3C CH3 H3Ci CH3 VCH3 CH3
269 270 271 272
CH3 H3C CH3 H3C,---CH3 CH3
Q1 Q1 Q1 Q1
H3C n CH3
H3C H3C CH3
273 274 275 276
CH3 ~CH3 -~CH3 H3C; CH3
Ql Qy Q1 Q1
CH3 C~ CH3 ~
CH3 CH3 CH3 H3C
277 278 279 280
CH3 "CH3 CH3
H3C ; CH3 =
Qi Q1 CH3 Q1 CH3 Ql ,,.CH3
~
H3C H3C H3C H3C
281 282 283 284
I-ICH3 CH3 I-ICH3
Q 1
Q1 .#CH3 Q1 CH3
Q1 ~
n3C HCH3
n
H3C H3C CH3 H3C CH3
285 286 287 288
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Table 2m
CH3
Q'~/~/~CH3 Q1 CH3 Q
Q1
CH3 CH3
~ CH3 H3
H3C CH3 CH3 CHs CH3
289 290 291 292
C_H3
CH3 H3C ~ -CH3
Q1 C Q1 H3 Q1H3C;CH3 Qi CH3
~ H3 LCH3
CH3 CH3 CH3
293 294 295 296
CH3 CH3 ,CH3 ~CH3
Q1 CH3 Ql - CH3 Q1,,,-YCH3 Q1-,-,,TCH3
CH3 CH3 CH3 CH3 CH3 CH3 CH3 CH3
297 298 299 300
H3C CH3 CH3 CH3 CH3
Q1 CH3 Q1 CH3 Q1' J_CH3 01 = CH3
CH H C CH3 ~v C~H3 H3
3 3 I'I3C H3C
301 302 303 304
C_H3 Q_ 1 CH3 Q1 CH3 Q1 CH3
Q1~CH3 _/I % T
= H3C CH CH H3C CH H3
H3C CH3 3 3 CHg 3 H3C CHV 3
305 306 307 308
Q1,,,/,ICH3 Q CH3 Q1\ ~rCH3 Q1 CH3
~
H3C CH3 CH3 H3C1 CH3 H3C CH CH3
3 H3C CH
3
309 310 311 312
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Table 2n
CH3 CH3 CH3 CH3 CH3 CH3 CH3 CH3
Q1- ~CH3 Q1~CH3 Q1~~f~CH3 Q1~~,/~CH3
CH3 CH3 CH3 CH3
313 314 315 316
CH3 Q1 CH3 Q1 CH3
CH3 C ~CH3
Q H3C ~~/\/~CH Q1 ~
~
3 CH3 CH3 CH3 CH3 CH3 CH3
317 318 319 320
CH3 CH3 CH3 CH3 CH3 CH3 CH3
Q1-,-~CH3 Q1CH3 Q1l"~CH3 Q1--~CH3
CH3 CH3 CH3 CH3 CH3
321 322 323 324
CH3 CH3 CH3 CH3 CH3
Q1-- j~~CH3 Q1~CH3 Q1~CH3 Q1 ~\ CH3
CH3 H3C CH3 H3C CH~H3 H3C CH~H3
325 326 327 328
H3C CH3 H3C CH3 CH3 CH3
Q1~CH3 Q1\~CH3 Q1~CH3 Q1~CH3
H3
CH3CH3
C_ Hs C1 H3 CH3CH3 H3 CH3 CH3
329 330 331 332
CH3 CH3 Q1 Q1 CH3
Q1~~CH Q1 =/~CCFi3 CH3 TrH
CH3 CH3 3 CH3 CH3 H3C CH3 H3C CH3 3
333 334 335 336
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Table 2o
Ql,,_,,-,,_iCH3 CH3 CH3 Qi CH3
= H3C CH3 H3C"I.-CH3
H3C C 3 H3 Q1 CH3 QlCH3 H3C CHCH3
3
337 338 339 340
Q~ CH CH3 CH3 H3C CH
Q1 CH3 Q1 CH3 Q1 eC CH3
HgC C 3 F-Ig H3C CH3 H3C C 3 H3 CH3
CH 3
3
41 342 343 344
3C CH3 Q1H3C CH3CH3 Q1 CH3 H3C CH3
Q1~CH3 \;~~ 1~\ 1'CH3 Ql CH3
CH3 H3C CH3 H3C CH3
CH 3 3 H3C CH3
3
345 346 347 348
HC CH3 H3C CH3 H C CH3 H C CH3
CH3 CH3 3 '~--CH3 3 ~--CH3
Q13 CH3 Q1 = CH3 Q"'-I'CH3 Q1~~CH3
CH3 CH3 CH3 CH3
349 350 351 352
H3C CH3 H3C CH3 CH3 CH3 CH3 CH3
Q1~CH3 Q1CH3 Q1 CH3 Q1 CH3
CH3 CH3 CH3 CH3
353 354 355 356
H3C CH3 H3C CH3 H3C
~ ~( H3C
Q1 XCH3 Q1'~i' V\CH3 Q1 Q1
CH3 CH3 CH3
357 358 359 360
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Table 2p
3H3C H3C CH3 1 CH3
Q1
Q1 * Q1 H3C7CH3
H3C CH3
CH3 CH3
361 362 363 364
CH3 CH3 CH3 3
Q1 *** CH3 Q1 CH Q1 CH3 Q1 CH
3 CH3
CH3 CH3 H3C*CH3 H3C CH3 *CH3
3
365 366 367 368
CH3
CH3 CH3 Q1 ** CH3 H3C H3C
Q1 *
C3
H3 Q1 Qi CH3 H3C *
CH3 CH3
369 370 371 372
H3C CH3 CH3
Q1 Q1 Q1 CH3 Q1 CH3
CH3 CH3 CH3 H3C CH3
CH3
373 374 375 376
CH3 CH3 CH3 H3C H3C CH3
Q1 * CH3 Q1 * * CH Q1 CH3 Q1
3
CH3 CH3 3C CH3 H3C CH3 H3C
H CH3 H3C CH3 CH3
CH3
377 378 379 380
CH3
Q1 H3C H3C H3C
H3C CH3 Q1 Q1 * Q1
CH3 CH3
381 382 383 384
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Table 2q
H3C CH3 CH3 CH3
Q~ * Qj Ql CH3 Q1 * CH3
CH3 CH3 )~CH3 CH3 CH3 CH3
385 386 387 388
CH3 CH3
H3CQ1 CH3 Q1 CH3 HsC H3C
H3C H3C CH3 H3 Q1 Q1
CH3 CH3
389 390 391 392
H3C CH3
H3C H3C
Q~ Q1 Q1 Q1
CH3
CH3 CH3 CH3
393 394 395 396
CH3 CH3 H3C CH3
CHg
CH3 CH3 CH3 CH3 Q CH3 Q1 H3C 3
Q1
rCH3
CH3 CH3 CH3 H3C
397 398 399 400
Q1'~'~CH3 CH3 Ql-"/ Q1~~~CH3
QJ/ CfH3
401 402 403 404
H3C
Q1 Q1~,CH3 Q1 \ Q1 /
CH3
405 406 407 408
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Table 2r
CH3
Q1 / CH3 Q,' /J CH3
C Y CH3 Q1 ~' CH3 Q1 j CH3
409 410 411 412
Q,\~~CH3 Q1 / CH3 Q1\~~CH3 Q1 n Ql
413 414 415 416
CH3 Q1\" '~ ~ /CH3
CH3 Q1~ Q1 \ CH3
417 418 419 420
CH3
Q, ~i~ CH3 H3C CH3 Q1~
CH3 Q 1 Y CH3 H3 _CI H3
421 422 423 424
CH3
3 ~ Q1 CH3
~
41\/)\\,CH3 Q1 ~ Q1
425 426 427 428
CH3 CH3 .CH3 Q1
Q, u1i a
d Q1~.
429 430 431 432
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Table 2s
Q Qi H3C CH3
Q~ Q1
CHs CH3
433 434 435 436
CH3 CH3 QH3
Ql .,."CH3
Q~\~r '' Q1 Q1\r'~
CH3
437 438 439 440
Q1~,,... H3 Q1 CH3 Qt%,,. CH3 CH3
3
CH3 '''
3 3 CH3
441 442 443 444
Q". CH3 CH3 : C H 3
CH3 Qi Q1u,,..
445 446 447 448
C H 3 CH3 CH3 CH3
Q Q 1 ...... Q,
CH3 CH3 CH3
449 450 451 452
Qy CH3 CH3 CH3
:?~ QI Q1nI...
453 454 455 456
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Table 2t
CH3 QH3
Q1~ Qinu..O Q1w--rCHg Q1~.-nIC~..~3
~~//
457 458 459 460
Q1 Q1 CH3
Q1-( ~ Q1
\J CH3
461 462 463 464
C_H3
Q1 0 1 Q1 CH3 Q1~~=,.,... CH3
~~
465 466 467 468
Q1\ .,\CH3 Qi~~r,..... .\\CH3 Q1 Q1
~' ~ CH3
H3C CH3 CH3
469 470 471 472
CH3 CH3 CH3
Q1~
CHg Q1 Q1 Q1
CH3 CH3 CH3 CH3
473 474 475 476
CH3
CH3 CH3 ,.\\CH3
Q1~ ~
= Q1 1\~~..'~ Q1'
CH3 CH3 CH3
CH3
477 478 479 480
- 68-
.__.___-_
, ----
CA 02268756 1999-04-15
WO 98/17647 PCTIUS97/18193
Table 2u
.,.~CH3 CH3 CH3 CH3
Q1\~''" Q1 Q1 r.'' ' ~
CH3 CH3 CH3 Q1 v -CH3
481 482 483 484
CH3
CH3 CH3
H3C CH3 H3Cx
Q1 . _ I X Q1\aP'H36CH3 H3C
CH3
485 486 487 488
Q1 .CH3 H3C CH3 H3C CH3 Qi
~ Q1 Q1m..
H3C CH3 H3C H3C CH3
489 490 491 492
Q1~ Q1 Q1 CH
= CH3 3 Q1\~~, CH3
CH3
493 494 495 496
Ql c Q1/i,... CH3
CH3 CH3
497 498 499 500
Q14,,,\CH3 Qi/i... CH3 Q1 CH3 Q14' CH3
CH3 CH3
501 502 503 504
- 69-
CA 02268756 1999-04-15
WO 98/17647 PCT/US97/18193
Table 2v
CH3 CH3
U CH3 A CH3 Q1
CH3 CH3 CH3 CH3
505 506 507 508
CH3 CH3 ,CH3 CH3
Q Q Q1 u..
, Q, jIf,..
CH3 ~!--CH3 <~-CH3 ~~-CH
3
509 510 511 512
,CH3 ,CH3 CH3 HgC
a Q1~, Q1 CH3
C ~' CH3 CH3 Q,
513 514 515 516
H3C H3C H3C
CH3 >-CH3 >-CH3 Q1
Q 1 0--< (QiuI...a *'--~
517 518 519 520
Q1 Q1 v 1
Q1
CH3 CH3 CH3 Q
CH3
521 522 523 524
Q1\v~''Q Q1\~ CH3
~ Q1\~.='Q
CH3 CH3 CH3 Q1~
525 526 527 528
- 70-
____.____,~__~_
, _--
CA 02268756 1999-04-15
WO 98/17647 PCT/US97/18193
Table 2w
CH3 CH3 CH3 CH3
Q1\a""~ Q1 Ql Q
'
H3C H3C H3C
529 530 531 532
CH3
H3C H3
Q1u,/.. QCHg Q1e ..,nCH3
H C Q
H3C 3 H3C
533 534 535 536
CH3 CH3 C H3
Q1~CH3 Q~ CH3 Q1ll-... CH3 01 - CH3
3 ~
537 538 539 540
CH3 CH3 CH3 CH3
Q1 ..mCHg Qlm-..' rCH3 Q1im.. .1111CH3 Q1mw-o=.-,iCH3
541 542 543 544
CH3 CH3 CH3
= Q' CHg
Q1i11,..O..n1CH3 Q1
545 546 547 548
~CH3 I--,CH3
~ CH3 CH3
Q1 Qiw...O Qi-( ~/ Ql~....m~
549 550 551 552
- 71-
__.._
CA 02268756 1999-04-15
WO 98/17647 PCTIUS97/18193
Table 2x
Q1 H3 H3C H3
Q1 Qimw~
Qii..
553 554 555 556
H3C H3C
CH3 CH3
Q1111-0 Q1~
557 558 559 560
,,CH3 Q1
Q1 ,.=CHs Qiu1,..= ~~~///
~ Q1
561 562 563 564
Q1 Q1\ Q1
1
Q
CH3
CH3 CH3 CH3
565 566 567 568
Q1\a"'' Q1", eo Q1 CH3
CH3 CH3 CH3
569 570 571 572
Q1 6 CH
Q1\r%,'"aCH3 Q1~=,,jCH3 Q
1\'a'' ~.,~jiCH3 3
573 574 575 576
- 72-
,
CA 02268756 1999-04-15
WO 98/17647 PCTIUS97/18193
Table 2y
Q1 =~CH3 Q1,,,, CH3 q1 5"XH3 H3 H3
H3 H3 ,,%CH3
577 578 579 580
Q 1 ,.,CH3 Q 1CH3 Q 1 ,oCH3 01 ~%,, CH3
CH3 CH3 s CH3 i~CH3
581 582 583 584
Q1 Q1n,.. Q1~ Q1115-0
CH3 CH3 \CH3 \CH
3
585 586 587 588
Q1 CH3 Q1u"... CH3 Q1 Q1u..
m--O.==,, ',, CH3
r,-CH3
589 590 591 592
CH3
Q1 Q1 Q1
Q1 =
CH3 CH3
593 594 595 596
CH3 CH3 CH3 CH3
Q1 Q1 Q1~~,r~=. Q1
597 598 599 600
- 73-
_
CA 02268756 1999-04-15
WO 98/17647 PCT/US97/18193
Table 2z
CH3
Q 1\/~=,,,. Q Q 1Q 1
CH3 O-dCH3 ~,~CH3
601 602 603 604
Q1\~~= 111CH3 Q1 Q1\,/\/ Q
1
CH3 CH3 CH
605 606 607 608
CH3
Q1\a~"Q1 Q~ V~! Q
RCH3 -CH3 -CH3
609 610 611 612
H3 H3 CH3 Ql
Q ,,,,~ ~=..,,,~
1\~v=='~ Q1\~ Q1\r~'
H3C
CH3
613 614 615 616
Q1~
Q1 u1-..0 Q1 -c - CH3
H3C CH3 H3C-\ CH3 H3C---\ CH3 H3C
617 618 619 620
Q ~ iw.. Q 1 w...
Q1
CH3 11,,,rCH3 CH3 Ql H3C H3C H3C CH3
621 622 623 624
- 74-
,
CA 02268756 1999-04-15
WO 98/17647 PCT/US97/18193
Table aa
Q1~ C1 Q1
CH3
,,,,,,CH3 H3C CH3 H3CCH3 Q1 CH3
625 626 627 628
CH3 Q1 CH3
~....,,, l
[D_~ < CH3
Q 1 \r'''. Q \
CH3 CH3 1\\,,.= CH3
629 630 631
CH3 CH3 ~-CH3
.n~
Q1 Q1\N''' Q1
632 633 634
-CH3 ~
C Q1
Q1~ Q1 *
Q1\r"
635 636 637 638
Q1 / Q1"~o Q1~ Q1 ~
639 640 641 642
H3C CH3
Q1
CH3 Q1
H3C *
643 644 645 646
- 75-
CA 02268756 1999-04-15
WO 98/17647 PCTIUS97/18193
Table 2ab
1H3C
Q Q1 * ~CH3
Q1 * ~
* a* Q1
CH3 H3C *
647 648 649 650
H3 CH3
*
Q1 D Q1 ~ Q1 CH3 Q1 ~
CH3
651 652 653 654
H3C CH3
Q1
Q1 Q1 bQ1 CH3
* CH3
655 656 657 658
H3 CHg Q1 \ /
Q
Q1
*\
659 660 661 662
Q1
Q1
Q1 \ I Q1 \ I ~~
CH 3 V
CH3 CH3
663 664 665 666
\ I \ I \ I / I
Q1 Q1 - Q1 Q1 \
CH3 CH3 H3C H3C
700 701 702 703
- 76-
~
CA 02268756 1999-04-15
WO 98/17647 PCT/US97/18193
Table 2ac
Q1 ( \ Q~ Ql \ Q1 ~ ~
CH3 CH3 CH3
i I
. ~
704 705 706 707
ZPI NH2 NH2
Q = Q7 Q1 Q1 NH2
CH3 CH3 CH3 CH3
708 709 710 711
OH , OH
~
Qj NH2 Qi-\ Q1~
CH3 CH3 ~CH3
712 713 714
OCH3
~I
Ql OH Q1 ~ OH Q1
CH3 CH3 CH3
715 716 717
OCH3 ' (
Q~ Qy OCH3 Q1 OCH3
CH3 CH3 CH3
718 719 720
F / F
Q1 Q1 Ql F Q1 F
CH3 '-CH3 CH3 CH3
721 722 723 724
- 77-
CA 02268756 1999-04-15
WO 98/17647 PCT/U597/18193
Table 3a
0 0 Q2 Q2
Q2-~/ Q24 OH ~O , CH3
OH O_CH3 ~ 0 0
a b c d
O~ //O O~ //O O 0
Q2 S,OH ~S,OH Q2" S, OH ~S,OH
Q2 Q2
e f 9 h
O 0 O 0S/O 0 0 0
~ = ~ ~ ~CH3 \
Q2 ~N CH3 Q2 N Q2 N S
CH3
H H H
i j k
O O O H H
Q2 " K N CH3 Q2-, SNYCH3 Q2~S'NyCH3
/
H 1' CH3 OO 0 O/ ~O O
m n
0
\\ OH O\ /0 - CH3 0 OH O\ O- CH3
p Q2 p F~ ~
Q2 ~OH ~ ~O _CH3 r ~OH r O-CH3
Q2 Q2
o p q r
- 78-
._ ----
------
CA 02268756 1999-04-15
WO 98/17647 PCT/US97/18193
Table 3b
H H H CH3 H CH3
N,,N N N QTS-I y Q2-, S.N -rf
O~ 0 \\ 0
N 1/N O~
Q2 O O
02
s t u v
Q2 Q2
H CH3 Q2
Q2-, . ~O ~O ~O
S ~CH3 p O O
p~ \
}-- CH3
p p H3C> H3C
CH3
w x y z
Q2 Q2 Q2 Q2
O O O O
O O O O
H3C-~ LCH3
CH3 CH3 CHg
H3C
A B C D
Q2 Q2
~=O \,~= O
O p
CH3 CH3
CH3
E F
- 79-
CA 02268756 1999-04-15
WO 98/17647 PCTIUS97/18193
Table 4a
Q3-OH Q3- N3 Q3- N02 Q3-NH2
1 2 3 4
Q3 NH2 Q3~ CH3 1 Q3
NH2 Q3/*' NH2
H2N
6 7 8
NH2 H H
Q3~/NH2 Q3~ Q3~N NH ~N~NH
T y CH3 CH3 NH2 Q3 NH2
9 10 11 12
H
NH NH
H3C\*'Ny NH Q3 NH
~ Q3)~ NH2 ~ NH2 L~,A NH2
Qg NH2 Q3
13 14 15 16
NH CH CH3
~S NH 3
H3C * NH2 Q3 y Q3~ NH2 Q " NH2
Q3 NH2
NH NH
17 18 19 20
NH2 NH2 NH2
~ CH3
~NH2 Q3NH2 --~
Q3 Q3 * NH2 Q3 ' I
NH2 NH2
21 22 23 24
- 80-
,
CA 02268756 1999-04-15
WO 98/17647 PCT/US97/18193
Table 4b
2
OH NH2
Q3 NHNH2 Q3 NH2 Q3~OH Q3NH2
OH
25 26 27 28
OH NH2 OH
Q3~OH ~ ~ * CH3
NH2 Q3 NH2 Q3!~/'~OH Q3
NH2
29 30 31 32
NH2 NH
CH3 NH2 Q3-CN ~ J(
43 * Q3 OH Q3 S NH2
OH
33 34 35 36
Q3
Q3 N~NH2 Q3~N y NH2 Q3 N y II NH2 ~,N NH2
~--
H CH3 SCH3 N
37 38 39 40
Q3 Q3 Q3
ro(S HN N~H
N~ y
I N~H
NH2 H' N\ CN H , N~
41 42 43
-81-
CA 02268756 1999-04-15
WO 98/17647 PCT/US97/18193
Table 4c
H
I H I H
Q3 N NCH3 Q~N NCH2CH3 I
NH2 3 NH Q3 N~CH3
2
44 45 46
H H H
I
Q3 N-,iCH3 Q3 N\/\CH3 Q3 N y CH3
H3C
47 48 49
H H CH3
Q3~ QH N Q3 Nl-~ NH2 Q3-" N-, CH3
50 51 52
(CH3
Q3-H
43 NN,iCH3
53 54
- 82-
CA 02268756 1999-04-15
WO 98/17647 PCT/US97/18193
Table 5a
~Q4 ~ 3
H- 04 H3C- Q4 H3C ~Q4 CH3 H3C Q4
a b c d e
0 0 0 H
H3C Q4 Q4 H3C--,A Q4 H3C N.Q
cl, '
~ 4
CH3 CH3 0
t 9 h
CH3 H CHs H H
' H3C\ /O, 1
N- Q4 7 Q4 H3C N, Q4 FH2CY N_Q4
O CH3 0 O
k I m
H H CH3 CH3 CH3
F2HCy N, Q4 F3Cy N, Q4 H3C N, Q
~ 4 N\Q4
O 0 0 O
n 0 p q
H 3 CH3 CH3 CH3
H3CyN, Q4 H3C CH N, Q4 FH2Cy N, Q4 F2HC N, Q4
CH3 0 O ~
0
r S t u
- 83-
CA 02268756 1999-04-15
WO 98/17647 PCT/US97/18193
Table 5b
CH3 H CH3 H CH3 H
F3Cy N, Q4 H3C, N, ~
~5~~ Q4 ;'S~ N, Q4 H3C S~ N, Q4
0 O O O~O O~ O
v w x y
CH3 CH3 CH3 CH3 CH3 H H
I' ~ N
H3C~ N. N
~S% Q4 S~ ~Q4 H3C' S~ N'Q4 ~N~Q
p O O,, O O ~O cN 4
z aa ab ac
H
iN N cINQ4 ~ 'Q4 N'Q4 .Q4
~ i H N
H H
H
ad ae af ag
H H
I I H
N' CD/r' N N\ N= N Q4 Q4 N ~ Q4 H
S
Q4
ah ai aj ak
H CH3 CH3
N~
N ~ N i I~N C1NQ4
~ Q4 N Q4 N.Q4 H CH3 CH3
al am an ao
-84-
CA 02268756 1999-04-15
WO 98/17647 PCT/US97/18193
Table 5c
a'~ CH3 CH3 CH3
I I
N,Q4 csN ~N'Q4 NN~N'Q
Q4 _S 4
CH3 S
ap aq ar as
H' H3C\ H\
H2N-Q4 N-Q4 N-Q4 N-Q4
H3C/ H3C/ H3C_/
at au av aw
H3C' 0,Q4 H3CN---p,Q4 H3C~S, Q4 H3C'I~S'Q4
ax ay az ba
0 0 0
H3C'K O' Q4 O Q4 H3C-)A OA4 S, Q4
CH3
CH3
bb bc bd be
0 p Sp 'N'
' Q4 Q4 N_"Q4
~
bf bg bh
O p Q4
N-Q4 N-Q4 CN0
bi bj bk
- 85-
CA 02268756 1999-04-15
WO 98/17647 PCT/US97/18193
Table 6 - Exemplary Enumerated Compounds
A.3.a.4.i; A.4.a.4.i; A.7.a.4.i; A.9.a.4.i; A.103.a.4.i; A.106.a.4.i;
A.107.a.4.i; A.108.a.4.i;
A.111.a.4.i; A.114.a.4.i; A.117.a.4.i; A.118.a.4.i; A.119.a.4.i; A.120.a.4.i;
A.121.a.4.i;
A.137.a.4.i; A.138.a.4.i; A.139.a.4.i; A.140.a.4.i; A.141.a.4.i; A.142.a.4.i;
A.145.a.4.i;
A.146.a.4.i; A.147.a.4.i; A.148.a.4.i; A.149.a.4.i; A.150.a.4.i; A.151.a.4.i;
A.165.a.4.i;
A.166.a.4.i; A.167.a.4.i; A.168.a.4.i; A.169.a.4.i; A.170.a.4.i; A.171.a.4.i;
A.172.a.4.i;
A.173.a.4.i; A.174.a.4.i; A.175.a.4.i; A.176.a.4.i; A.188.a.4.i; A.189.a.4.i;
A.190.a.4.i;
A.196.a.4.i; A.202.a.4.i; A.205.a.4.i; A.206.a.4.i; A.207.a.4.i; A.208.a.4.i;
A.209.a.4.i;
A.210.a.4.i; A.211.a.4.i; A.212.a.4.i; A.213.a.4.i; A.700.a.4.i; A.701.a.4.i;
A.702.a.4.i;
A.703.a.4.i; A.704.a.4.i; A.705.a.4.i; A.706.a.4.i; A.707.a.4.i; A.708.a.4.i;
A.709.a.4.i;
A.710.a.4.i; A.711.a.4.i; A.712.a.4.i; A.713.a.4.i; A.714.a.4.i; A.715.a.4.i;
A.716.a.4.i;
A.717.a.4.i; A.718.a.4.i; A.719.a.4.i; A.720.a.4.i; A.721.a.4.i; A.722.a.4.i;
A.723.a.4.i;
A.724.a.4.i; A.3.a.4.o; A.4.a.4.o; A.7.a.4.o; A.9.a.4.o; A.103.a.4.o;
A.106.a.4.o;
A.107.a.4.o; A.108.a.4.o; A.111.a.4.o; A.114.a.4.o; A.117.a.4.o; A.118.a.4.o;
A.119.a.4.o; A.120.a.4.o; A.121.a.4.o; A.137.a.4.o; A.138.a.4.o; A.139.a.4.o;
A.140.a.4.o; A.141.a.4.o; A.142.a.4.o; A.145.a.4.o; A.146.a.4.o; A.147.a.4.o;
A.148.a.4.o; A.149.a.4.o; A.150.a.4.o; A.151.a.4.o; A.165.a.4.o; A.166.a.4.o;
A.167.a.4.o; A.168.a.4.o; A.169.a.4.o; A.170.a.4.o; A.171.a.4.o; A.172.a.4.o;
A.173.a.4.o; A.174.a.4.o; A.175.a.4.o; A.176.a.4.o; A.188.a.4.o; A.189.a.4.o;
A.190.a.4.o; A.196.a.4.o; A.202.a.4.o; A.205.a.4.o; A.206.a.4.o; A.207.a.4.o;
A.208.a.4.o; A.209.a.4.o; A.210.a.4.o; A.211.a.4.o; A.212.a.4.o; A.213.a.4.o;
A.700.a.4.o; A.701.a.4.o; A.702.a.4.o; A.703.a.4.o; A.704.a.4.o; A.705.a.4.o;
A.706.a.4.o; A.707.a.4.o; A.708.a.4.o; A.709.a.4.o; A.710.a.4.o; A.711.a.4.o;
A.712.a.4.o; A.713.a.4.o; A.714.a.4.o; A.715.a.4.o; A.716.a.4.o; A.717.a.4.o;
A.718.a.4.o; A.719.a.4.o; A.720.a.4.o; A.721.a.4.o; A.722.a.4.o; A.723.a.4.o;
A.724.a.4.o; A.172.b.4.i; A.173.b.4.i; A.174.b.4.i; A.175.b.4.i; A.176.b.4.i;
A.188.b.4.i;
A.189.b.4.i; A.190.b.4.i; A.196.b.4.i; A.202.b.4.i; A.205.b.4.i; A.206.b.4.i;
A.207.b.4.i;
A.208.b.4.i; A.209.b.4.i; A.210.b.4.i; A.211.b.4.i; A.212.b.4.i; A.213.b.4.i;
A.700.b.4.i;
A.701.b.4.i; A.702.b.4.i; A.703.b.4.i; A.704.b.4.i; A.705.b.4.i; A.706.b.4.i;
A.707.b.4.i;
A.708.b.4.i; A.709.b.4.i; A.710.b.4.i; A.711.b.4.i; A.712.b.4.i; A.713.b.4.i;
A.714.b.4.i;
A.715.b.4.i; A.716.b.4.i; A.717.b.4.i; A.718.b.4.i; A.719.b.4.i; A.720.b.4.i;
A.721.b.4.i;
A.722.b.4.i; A.723.b.4.i; A.724.b.4.i; A.3.b.4.o; A.4.b.4.o; A.7.b.4.o;
A.9.b.4.o;
A.103.b.4.o; A.106.b.4.o; A.107.b.4.o; A.108.b.4.o; A.111.b.4.o; A.114.b.4.o;
A.117.b.4.o; A.118.b.4.o; A.119.b.4.o; A.120.b.4.o; A.121.b.4.o; A.137.b.4.o;
A.138.b.4.o; A.139.b.4.o; A.140.b.4.o; A.141.b.4.o; A.142.b.4.o; A.145.b.4.o;
A.146.b.4.o; A.147.b.4.o; A.148.b.4.o; A.149.b.4.o; A.150.b.4.o; A.151.b.4.o;
A.165.b.4.o; A.166.b.4.o; A.167.b.4.o; A.168.b.4.o; A.169.b.4.o; A.170.b.4.o;
A.171.b.4.o; A.172.b.4.o; A.173.b.4.o; A.174.b.4.o; A.175.b.4.o; A.176.b.4.o;
A.188.b.4.o; A.189.b.4.o; A.190.b.4.o; A.196.b.4.o; A.202.b.4.o; A.205.b.4.o;
A.206.b.4.o; A.207.b.4.o; A.208.b.4.o; A.209.b.4.o; A.210.b.4.o; A.211.b.4.o;
A.212.b.4.o; A.213.b.4.o; A.700.b.4.o; A.701.b.4.o; A.702.b.4.o; A.703.b.4.o;
A.704.b.4.o; A.705.b.4.o; A.706.b.4.o; A.707.b.4.o; A.708.b.4.o; A.709.b.4.o;
A.710.b.4.o; A.711.b.4.o; A.712.b.4.o; A.713.b.4.o; A.714.b.4.o; A.715.b.4.o;
A.716.b.4.o; A.717.b.4.o; A.718.b.4.o; A.719.b.4.o; A.720.b.4.o; A.721.b.4.o;
A.722.b.4.o; A.723.b.4.o; A.724.b.4.o; A.172.x.4.i; A.173.x.4.i; A.174.x.4.i;
A.175.x.4.i;
A.176.x.4.i; A.188.x.4.i; A.189.x.4.i; A.190.x.4.i; A.196.x.4.i; A.202.x.4.i;
A.205.x.4.i;
A.206.x.4.i; A.207.x.4.i; A.208.x.4.i; A.209.x.4.i; A.210.x.4.i; A.211.x.4.i;
A.212.x.4.i;
A.213.x.4.i; A.700.x.4.i; A.701.x.4.i; A.702.x.4.i; A.703.x.4.i; A.704.x.4.i;
A.705.x.4.i;
A.706.x.4.i; A.707.x.4.i; A.708.x.4.i; A.709.x.4.i; A.710.x.4.i; A.711.x.4.i;
A.712.x.4.i;
A.713.x.4.i; A.714.x.4.i; A.715.x.4.i; A.716.x.4.i; A.717.x.4.i; A.718.x.4.i;
A.719.x.4.i;
A.720.x.4.i; A.721.x.4.i; A.722.x.4.i; A.723.x.4.i; A.724.x.4.i; A.3.x.4.o;
A.4.x.4.o;
A.7.x.4.o; A.9.x.4.o; A.103.x.4.o; A.106.x.4.o; A.107.x.4.o; A.108.x.4.o;
A.111.x.4.o;
A.114.x.4.o; A.117.x.4.o; A.118.x.4.o; A.119.x.4.o; A.120.x.4.o; A.121.x.4.o;
A.137.x.4.o; A.138.x.4.o; A.139.x.4.o; A.140.x.4.o; A.141.x.4.o; A.142.x.4.o;
A.145.x.4.o; A.146.x.4.o; A.147.x.4.o; A.148.x.4.o; A.149.x.4.o; A.150.x.4.o;
- 86-
CA 02268756 1999-04-15
WO 98/17647 PCT/US97/18193
A.151.x.4.o; A.165.x.4.o; A.166.x.4.o; A.167.x.4.o; A.168.x.4.o; A.169.x.4.o;
A.170.x.4.o; A.171.x.4.o; A.172.x.4.o; A.173.x.4.o; A.174.x.4.o; A.175.x.4.o;
A.176.x.4.o; A.188.x.4.o; A.189.x.4.o; A.190.x.4.o; A.196.x.4.o; A.202.x.4.o;
A.205.x.4.o; A.206.x.4.o; A.207.x.4.o; A.208.x.4.o; A.209.x.4.o; A.210.x.4.o;
A.211.x.4.o; A.212.x.4.o; A.213.x.4.o; A.700.x.4.o; A.701.x.4.o; A.702.x.4.o;
A.703.x.4.o; A.704.x.4.o; A.705.x.4.o; A.706.x.4.o; A.707.x.4.o; A.708.x.4.o;
A.709.x.4.o; A.710.x.4.o; A.711.x.4.o; A.712.x.4.o; A.713.x.4.o; A.714.x.4.o;
A.715.x.4.o; A.716.x.4.o; A.717.x.4.o; A.718.x.4.o; A.719.x.4.o; A.720.x.4.o;
A.721.x.4.o; A.722.x.4.o; A.723.x.4.o; A.724.x.4.o; A.172.y.4.i; A.173.y.4.i;
A.174.y.4.i;
A.175.y.4.i; A.176.y.4.i; A.188.y.4.i; A.189.y.4.i; A.190.y.4.i; A.196.y.4.i;
A.202.y.4.i;
A.205.y.4.i; A.206.y.4.i; A.207.y.4.i; A.208.y.4.i; A.209.y.4.i; A.210.y.4.i;
A.211.y.4.i;
A.212.y.4.i; A.213.y.4.i; A.700.y.4.i; A.701.y.4.i; A.702.y.4.i; A.703.y.4.i;
A.704.y.4.i;
A.705.y.4.i; A.706.y.4.i; A.707.y.4.i; A.708.y.4.i; A.709.y.4.i; A.710.y.4.i;
A.711.y.4.i;
A.712.y.4.i; A.713.y.4.i; A.714.y.4.i; A.715.y.4.i; A.716.y.4.i; A.717.y.4.i;
A.718.y.4.i;
A.719.y.4.i; A.720.y.4.i; A.721.y.4.i; A.722.y.4.i; A.723.y.4.i; A.724.y.4.i;
A.3.y.4.o;
A.4.y.4.o; A.7.y.4.o; A.9.y.4.o; A.103.y.4.o; A.106.y.4.o; A.107.y.4.o;
A.108.y.4.o;
A.111.y.4.o; A.114.y.4.o; A.117.y.4.o; A.118.y.4.o; A.119.y.4.o; A.120.y.4.o;
A.121.y.4.o; A.137.y.4.o; A.138.y.4.o; A.139.y.4.o; A.140.y.4.o; A.141.y.4.o;
A.142.y.4.o; A.145.y.4.o; A.146.y.4.o; A.147.y.4.o; A.148.y.4.o; A.149.y.4.o;
A.150.y.4.o; A.151.y.4.o; A.165.y.4.o; A.166.y.4.o; A.167.y.4.o; A.168.y.4.o;
A.169.y.4.o; A.170.y.4.o; A.171.y.4.o; A.172.y.4.o; A.173.y.4.o; A.174.y.4.o;
A.175.y.4.o; A.176.y.4.o; A.188.y.4.o; A.189.y.4.o; A.190.y.4.o; A.196.y.4.o;
A.202.y.4.o; A.205.y.4.o; A.206.y.4.o; A.207.y.4.o; A.208.y.4.o; A.209.y.4.o;
A.210.y.4.o; A.211.y.4.o; A.212.y.4.o; A.213.y.4.o; A.700.y.4.o; A.701.y.4.o;
A.702.y.4.o; A.703.y.4.o; A.704.y.4.o; A.705.y.4.o; A.706.y.4.o; A.707.y.4.o;
A.708.y.4.o; A.709.y.4.o; A.710.y.4.o; A.711.y.4.o; A.712.y.4.o; A.713.y.4.o;
A.714.y.4.o; A.715.y.4.o; A.716.y.4.o; A.717.y.4.o; A.718.y.4.o; A.719.y.4.o;
A.720.y.4.o; A.721.y.4.o; A.722.y.4.o; A.723.y.4.o; A.724.y.4.o; A.172.z.4.i;
A.173.z.4.i;
A.174.z.4.i; A.175.z.4.i; A.176.z.4.i; A.188.z.4.i; A.189.z.4.i; A.190.z.4.i;
A.196.z.4.i;
A.202.z.4.i; A.205.z.4.i; A.206.z.4.i; A.207.z.4.i; A.208.z.4.i; A.209.z.4.i;
A.210.z.4.i;
A.211.z.4.i; A.212.z.4.i; A.213.z.4.i; A.700.z.4.i; A.701.z.4.i; A.702.z.4.i;
A.703.z.4.i;
A.704.z.4.i; A.705.z.4.i; A.706.z.4.i; A.707.z.4.i; A.708.z.4.i; A.709.z.4.i;
A.710.z.4.i;
A.711.z.4.i; A.712.z.4.i; A.713.z.4.i; A.714.z.4.i; A.715.z.4.i; A.716.z.4.i;
A.717.z.4.i;
A.718.z.4.i; A.719.z.4.i; A.720.z.4.i; A.721.z.4.i; A.722.z.4.i; A.723.z.4.i;
A.724.z.4.i;
A.3.z.4.o; A.4.z.4.o; A.7.z.4.o; A.9.z.4.o; A.103.z.4.o; A.106.z.4.o;
A.107.z.4.o;
A.108.z.4.o; A.111.z.4.o; A.114.z.4.o; A.117.z.4.o; A.118.z.4.o; A.119.z.4.o;
A.120.z.4.o; A.121.z.4.o; A.137.z.4.o; A.138.z.4.o; A.139.z.4.o; A.140.z.4.o;
A.141.z.4.o; A.142.z.4.o; A.145.z.4.o; A.146.z.4.o; A.147.z.4.o; A.148.z.4.o;
A.149.z.4.o; A.150.z.4.o; A.151.z.4.o; A.165.z.4.o; A.166.z.4.o; A.167.z.4.o;
A.168.z.4.o; A.169.z.4.o; A.170.z.4.o; A.171.z.4.o; A.172.z.4.o; A.173.z.4.o;
A.174.z.4.o; A.175.z.4.o; A.176.z.4.o; A.188.z.4.o; A.189.z.4.o; A.190.z.4.o;
A.196.z.4.o; A.202.z.4.o; A.205.z.4.o; A.206.z.4.o; A.207.z.4.o; A.208.z.4.o;
A.209.z.4.o; A.210.z.4.o; A.211.z.4.o; A.212.z.4.o; A.213.z.4.o; A.700.z.4.o;
A.701.z.4.o; A.702.z.4.o; A.703.z.4.o; A.704.z.4.o; A.705.z.4.o; A.706.z.4.o;
A.707.z.4.o; A.708.z.4.o; A.709.z.4.o; A.710.z.4.o; A.711.z.4.o; A.712.z.4.o;
A.713.z.4.o; A.714.z.4.o; A.715.z.4.o; A.716.z.4.o; A.717.z.4.o; A.718.z.4.o;
A.719.z.4.o; A.720.z.4.o; A.721.z.4.o; A.722.z.4.o; A.723.z.4.o; A.724.z.4.o;
A.172.A.4.i; A.173.A.4.i; A.174.A.4.i; A.175.A.4.i; A.176.A.4.i; A.188.A.4.i;
A.189.A.4.i; A.190.A.4.i; A.196.A.4.i; A.202.A.4.i; A.205.A.4.i; A.206.A.4.i;
A.207.A.4.i; A.208.A.4.i; A.209.A.4.i; A.210.A.4.i; A.211.A.4.i; A.212.A.4.i;
A.213.A.4.i; A.700.A.4.i; A.701.A.4.i; A.702.A.4.i; A.703.A.4.i; A.704.A.4.i;
A.705.A.4.i; A.706.A.4.i; A.707.A.4.i; A.708.A.4.i; A.709.A.4.i; A.710.A.4.i;
A.711.A.4.i; A.712.A.4.i; A.713.A.4.i; A.714.A.4.i; A.715.A.4.i; A.716.A.4.i;
A.717.A.4.i; A.718.A.41; A.719.A.4.i; A.720.A.4.i; A.721.A.4.i; A.722.A.4.i;
A.723.A.4.i; A.724.A.4.i; A.3.A.4.o; A.4.A.4.o; A.7.A.4.o; A.9.A.4.o;
A.103.A.4.o;
A.106.A.4.o; A.107.A.4.o; A.108.A.4.o; A.111.A.4.o; A.114.A.4.o; A.117.A.4.o;
- 87-
__..__
CA 02268756 1999-04-15
WO 98/17647 PCT/US97/18193
A.118.A.4.o; A.119.A.4.o; A.120.A.4.o; A.121.A.4.o; A.137.A.4.o; A.138.A.4.o;
A.139.A.4.o; A.140.A.4.o; A.141.A.4.o; A.142.A.4.o; A.145.A.4.o; A.146.A.4.o;
A.147.A.4.o; A.148.A.4.o; A.149.A.4.o; A.150.A.4.o; A.151.A.4.o; A.165.A.4.o;
A.166.A.4.o; A.167.A.4.o; A.168.A.4.o; A.169.A.4.o; A.170.A.4.o; A.171.A.4.o;
A.172.A.4.o; A.173.A.4.o; A.174.A.4.o; A.175.A.4.o; A.176.A.4.o; A.188.A.4.o;
A.189.A.4.o; A.190.A.4.o; A.196.A.4.o; A.202.A.4.o; A.205.A.4.o; A.206.A.4.o;
A.207.A.4.o; A.208.A.4.o; A.209.A.4.o; A.210.A.4.o; A.211.A.4.o; A.212.A.4.o;
A.213.A.4.o; A.700.A.4.o; A.701.A.4.o; A.702.A.4.o; A.703.A.4.o; A.704.A.4.o;
A.705.A.4.o; A.706.A.4.o; A.707.A.4.o; A.708.A.4.o; A.709.A.4.o; A.710.A.4.o;
A.711.A.4.o; A.712.A.4.o; A.713.A.4.o; A.714.A.4.o; A.715.A.4.o; A.716.A.4.o;
A.717.A.4.o; A.718.A.4.o; A.719.A.4.o; A.720.A.4.o; A.721.A.4.o; A.722.A.4.o;
A.723.A.4.o; A.724.A.4.o; A.172.B.4.i; A.173.B.4.i; A.174.B.4.i; A.175.B.4.i;
A.176.B.4.i;
A.188.B.4.i; A.189.B.4.i; A.190.B.4.i; A.196.B.4.i; A.202.B.4.i; A.205.B.4.i;
A.206.B.4.i;
A.207.B.4.i; A.208.B.4.i; A.209.B.4.i; A.210.B.4.i; A.211.B.4.i; A.212.B.4.i;
A.213.B.4.i;
A.700.B.4.i; A.701.B.4.i; A.702.B.4.i; A.703.B.4.i; A.704.B.4.i; A.705.B.4.i;
A.706.B.4.i;
A.707.B.4.i; A.708.B.4.i; A.709.B.4.i; A.710.B.4.i; A.711.B.4.i; A.712.B.4.i;
A.713.B.4.i;
A.714.B.4.i; A.715.B.4.i; A.716.B.4.i; A.717.B.4.i; A.718.B.4.i; A.719.B.4.i;
A.720.B.4.i;
A.721.B.4.i; A.722.B.4.i; A.723.B.4.i; A.724.B.4.i; A.3.B.4.o; A.4.B.4.o;
A.7.B.4.o;
A.9.B.4.o; A.103.B.4.o; A.106.B.4.o; A.107.B.4.o; A.108.B.4.o; A.111.B.4.o;
A.114.B.4.o;
A.117.B.4.o; A.118.B.4.o; A.119.B.4.o; A.120.B.4.o; A.121.B.4.o; A.137.B.4.o;
A.138.B.4.o; A.139.B.4.o; A.140.B.4.o; A.141.B.4.o; A.142.B.4.o; A.145.B.4.o;
A.146.B.4.o; A.147.B.4.o; A.148.B.4.o; A.149.B.4.o; A.150.B.4.o; A.151.B.4.o;
A.165.B.4.o; A.166.B.4.o; A.167.B.4.o; A.168.B.4.o; A.169.B.4.o; A.170.B.4.o;
A.171.B.4.o; A.172.B.4.o; A.173.B.4.o; A.174.B.4.o; A.175.B.4.o; A.176.B.4.o;
A.188.B.4.o; A.189.B.4.o; A.190.B.4.o; A.196.B.4.o; A.202.B.4.o; A.205.B.4.o;
A.206.B.4.o; A.207.B.4.o; A.208.B.4.o; A.209.B.4.o; A.210.B.4.o; A.211.B.4.o;
A.212.B.4.o; A.213.B.4.o; A.700.B.4.o; A.701.B.4.o; A.702.B.4.o; A.703.B.4.o;
A.704.B.4.o; A.705.B.4.o; A.706.B.4.o; A.707.B.4.o; A.708.B.4.o; A.709.B.4.o;
A.710.B.4.o; A.711.B.4.o; A.712.B.4.o; A.713.B.4.o; A.714.B.4.o; A.715.B.4.o;
A.716.B.4.o; A.717.B.4.o; A.718.B.4.o; A.719.B.4.o; A.720.B.4.o; A.721.B.4.o;
A.722.B.4.o; A.723.B.4.o; A.724.B.4.o; B.3.a.54.i; B.4.a.54.i; B.7.a.54.i;
B.9.a.54.i;
B.103.a.54.i; B.106.a.54.i; B.107.a.54.i; B.108.a.54.i; B.111.a.54.i;
B.114.a.54.i;
B.117.a.54.i; B.118.a.54.i; B.119.a.54.i; B.120.a.54.i; B.121.a.54.i;
B.137.a.54.i;
B.138.a.54.i; B.139.a.54.i; B.140.a.54.i; B.141.a.54.i; B.142.a.54.i;
B.145.a.54.i;
B:146.a.54.i; B.147.a.54.i; B.148.a.54.i; B.149.a.54.i; B.150.a.54.i;
B.151.a.54.i;
B.165.a.54.i; B.166.a.54.i; B.167.a.54.i; B.168.a.54.i; B.169.a.54.i;
B.170.a.54.i;
B.171.a.54.i; B.172.a.54.i; B.173.a.54.i; B.174.a.54.i; B.175.a.54.i;
B.176.a.54.i;
B:188.a.54.i; B.189.a.54.i; B.190.a.54.i; B.196.a.54.i; B.202.a.54.i;
B.205.a.54.i;
B.206.a.54.i; B.207.a.54.i; B.208.a.54.i; B.209.a.54.i; B.210.a.54.i;
B.211.a.54.i;
B.212.a.54.i; B.213.a.54.i; B.700.a.54.i; B.701.a.54.i; B.702.a.54.i;
B.703.a.54.i;
B.704.a.54.i; B.705.a.54.i; B.706.a.54.i; B.707.a.54.i; B.708.a.54.i;
B.709.a.54.i;
B.710.a.54.i; B.711.a.54.i; B.712.a.54.i; B.713.a.54.i; B.714.a.54.i;
B.715.a.54.i;
B.716.a.54.i; B.717.a.54.i; B.718.a.54.i; B.719.a.54.i; B.720.a.54.i;
B.721.a.54.i;
B.722.a.54.i; B.723.a.54.i; B.724.a.54.i; B.3.a.54.o; B.4.a.54.o; B.7.a.54.o;
B.9.a.54.o;
B.103.a.54.o; B.106.a.54.o; B.107.a.54.o; B.108.a.54.o; B.111.a.54.o;
B.114.a.54.o;
B.117.a.54.o; B.118.a.54.o; B.119.a.54.o; B.120.a.54.o; B.121.a.54.o;
B.137.a.54.o;
B.138.a.54.o; B.139.a.54.o; B.140.a.54.o; B.141.a.54.o; B.142.a.54.o;
B.145.a.54.o;
B.146.a.54.o; B.147.a.54.o; B.148.a.54.o; B.149.a.54.o; B.150.a.54.o;
B.151.a.54.o;
B.165.a.54.o; B.166.a.54.o; B.167.a.54.o; B.168.a.54.o; B.169.a.54.o;
B.170.a.54.o;
B.171.a.54.o; B.172.a.54.o; B.173.a.54.o; B.174.a.54.o; B.175.a.54.o;
B.176.a.54.o;
B.188.a.54.o; B.189.a.54.o; B.190.a.54.o; B.196.a.54.o; B.202.a.54.o;
B.205.a.54.o;
B.206.a.54.o; B.207.a.54.o; B.208.a.54.o; B.209.a.54.o; B.210,a.54.o;
B.211.a.54.o;
B.212.a.54.o; B.213.a.54.o; B.700.a.54.o; B.701.a.54.o; B.702.a.54.o;
B.703.a.54.o;
B.704.a.54.o; B.705.a.54.o; B.706.a.54.o; B.707.a.54.o; B.708.a.54.o;
B.709.a.54.o;
B.710.a.54.o; B.711.a.54.o; B.712.a.54.o; B.713.a.54.o; B.714.a.54.o;
B.715.a.54.o;
B.716.a.54.o; B.717.a.54.o; B.718.a.54.o; B.719.a.54.o; B.720.a.54.o;
B.721.a.54.o;
- 88-
CA 02268756 1999-04-15
WO 98/17647 PCT/US97/18193
B.722.a.54.o; B.723.a.54.o; B.724.a.54.o; B.172.b.54.i; B.173.b.54.i;
B.174.b.54.i;
B.175.b.54.i; B.176.b.54.i; B.188.b.54.i; B.189.b.54.i; B.190.b.54.i;
B.196.b.54.i;
B.202.b.54.i; B.205.b.54.i; B.206.b.54.i; B.207.b.54.i; B.208.b.54.i;
B.209.b.54.i;
B.210.b.54.i; B.211.b.54.i; B.212.b.54.i; B.213.b.54.i; B.700.b.54.i;
B.701.b.54.i;
B.702.b.54.i; B.703.b.54.i; B.704.b.54.i; B.705.b.54.i; B.706.b.54.i;
B.707.b.54.i;
B.708.b.54.i; B.709.b.54.i; B.710.b.54.i; B.711.b.54.i; B.712.b.54.i;
B.713.b.54.i;
B.714.b.54.i; B.715.b.54.i; B.716.b.54.i; B.717.b.54.i; B.718.b.54.i;
B.719.b.54.i;
B.720.b.54.i; B.721.b.54.i; B.722.b.54.i; B.723.b.54.i; B.724.b.54.i;
B.3.b.54.o; B.4.b.54.o;
B.7.b.54.o; B.9.b.54.o; B.103.b.54.o; B.106.b.54.o; B.107.b.54.o;
B.108.b.54.o;
B.111.b.54.o; B.114.b.54.o; B.117.b.54.o; B.118.b.54.o; B.119.b.54.o;
B.120.b.54.o;
B.121.b.54.o; B.137.b.54.o; B.138.b.54.o; B.139.b.54.o; B.140.b.54.o;
B.141.b.54.o;
B.142.b.54.o; B.145.b.54.o; B.146.b.54.o; B.147.b.54.o; B.148.b.54.o;
B.149.b.54.o;
B.150.b.54.o; B.151.b.54.o; B.165.b.54.o; B.166.b.54.o; B.167.b.54.o;
B.168.b.54.o;
B.169.b.54.o; B.170.b.54.o; B.171.b.54.o; B.172.b.54.o; B.173.b.54.o;
B.174.b.54.o;
B.175.b.54.o; B.176.b.54.o; B.188.b.54.o; B.189.b.54.o; B.190.b.54.o;
B.196.b.54.o;
B.202.b.54.o; B.205.b.54.o; B.206.b.54.o; B.207.b.54.o; B.208.b.54.o;
B.209.b.54.o;
B.210.b.54.o; B.211.b.54.o; B.212.b.54.o; B.213.b.54.o; B.700.b.54.o;
B.701.b.54.o;
B.702.b.54.o; B.703.b.54.o; B.704.b.54.o; B.705.b.54.o; B.706.b.54.o;
B.707.b.54.o;
B.708.b.54.o; B.709.b.54.o; B.710.b.54.o; B.711.b.54.o; B.712.b.54.o;
B.713.b.54.o;
B.714.b.54.o; B.715.b.54.o; B.716.b.54.o; B.717.b.54.o; B.718.b.54.o;
B.719.b.54.o;
B.720.b.54.o; B.721.b.54.o; B.722.b.54.o; B.723.b.54.o; B.724.b.54.o;
B.172.x.54.i;
B.173.x.54.i; B.174.x.54.i; B.175.x.54.i; B.176.x.54.i; B.188.x.54.i;
B.189.x.54.i;
B.190.x.54.i; B.196.x.54.i; B.202.x.54.i; B.205.x.54.i; B.206.x.54.i;
B.207.x.54.i;
B.208.x.54.i; B.209.x.54.i; B.210.x.54.i; B.211.x.54.i; B.212.x.54.i;
B.213.x.54.i;
B.700.x.54.i; B.701.x.54.i; B.702.x.54.i; B.703.x.54.i; B.704.x.54.i;
B.705.x.54.i;
B.706.x.54.i; B.707.x.54.i; B.708.x.54.i; B.709.x.54.i; B.710.x.54.i;
B.711.x.54.i;
B.712.x.54.i; B.713.x.54.i; B.714.x.54.i; B.715.x.54.i; B.716.x.54.i;
B.717.x.54.i;
B.718.x.54.i; B.719.x.54.i; B.720.x.54.i; B.721.x.54.i; B.722.x.54.i;
B.723.x.54.i;
B.724.x.54.i; B.3.x.54.o; B.4.x.54.o; B.7.x.54.o; B.9.x.54.o; B.103.x.54.o;
B.106.x.54.o;
B.107.x.54.o; B.108.x.54.o; B.111.x.54.o; B.114.x.54.o; B.117.x.54.o;
B.118.x.54.o;
B.119.x.54.o; B.120.x.54.o; B.121.x.54.o; B.137.x.54.o; B.138.x.54.o;
B.139.x.54.o;
B.140.x.54.o; B.141.x.54.o; B.142.x.54.o; B.145.x.54.o; B.146.x.54.o;
B.147.x.54.o;
B.148.x.54.o; B.149.x.54.o; B.150.x.54.o; B.151.x.54.o; B.165.x.54.o;
B.166.x.54.o;
B.167.x.54.o; B.168.x.54.o; B.169.x.54.o; B.170.x.54.o; B.171.x.54.o;
B.172.x.54.o;
B.173.x.54.o; B.174.x.54.o; B.175.x.54.o; B.176.x.54.o; B.188.x.54.o;
B.189.x.54.o;
B.190.x.54.o; B.196.x.54.o; B.202.x.54.o; B.205.x.54.o; B.206.x.54.o;
B.207.x.54.o;
B.208.x.54.o; B.209.x.54.o; B.210.x.54.o; B.211.x.54.o; B.212.x.54.o;
B.213.x.54.o;
B.700.x.54.o; B.701.x.54.o; B.702.x.54.o; B.703.x.54.o; B.704.x.54.o;
B.705.x.54.o;
B.706.x.54.o; B.707.x.54.o; B.708.x.54.o; B.709.x.54.o; B.710.x.54.o;
B.711.x.54.o;
B.712.x.54.o; B.713.x.54.o; B.714.x.54.o; B.715.x.54.o; B.716.x.54.o;
B.717.x.54.o;
B.718.x.54.o; B.719.x.54.o; B.720.x.54.o; B.721.x.54.o; B.722.x.54.o;
B.723.x.54.o;
B.724.x.54.o; B.172.y.54.i; B.173.y.54.i; B.174.y.54.i; B.175.y.54.i;
B.176.y.54.i;
B.188.y.54.i; B.189.y.54.i; B.190.y.54.i; B.196.y.54.i; B.202.y.54.i;
B.205.y.54.i;
B.206:y.54.i; B.207.y.54.i; B.208.y.54.i; B.209.y.54.i; B.210.y.54.i;
B.211.y.54.i;
B.212.y.54.i; B.213.y.54.i; B.700.y.54.i; B.701.y.54.i; B.702.y.54.i;
B.703.y.54.i;
B.704.y.54.i; B.705.y.54.i; B.706.y.54.i; B.707.y.54.i; B.708.y.54.i;
B.709.y.54.i;
B.710.y.54.i; B.711.y.54.i; B.712.y.54.i; B.713.y.54.i; B.714.y.54.i;
B.715.y.54.i;
B.716.y.54.i; B.717.y.54.i; B.718.y.54.i; B.719.y.54.i; B.720.y.54.i;
B.721.y.54.i;
B.722.y.54.i; B.723.y.54.i; B.724.y.54.i; B.3.y.54.o; B.4.y.54.o; B.7.y.54.o;
B.9.y.54.o;
B.103.y.54.o; B.106.y.54.o; B.107.y.54.o; B.108.y.54.o; B.111.y.54.o;
B.114.y.54.o;
B.117.y.54.o; B.118.y.54.o; B.119.y.54.o; B.120.y.54.o; B.121.y.54.o;
B.137.y.54.o;
, B.138.y.54.o; B.139.y.54.o; B.140.y.54.o; B.141.y.54.o; B.142.y.54.o;
B.145.y.54.o;
B.146.y.54.o; B.147.y.54.o; B.148.y.54.o; B.149.y.54.o; B.150.y.54.o;
B.151.y.54.o;
B.165.y.54.o; B.166.y.54.o; B.167.y.54.o; B.168.y.54.o; B.169.y.54.o;
B.170.y.54.o;
B.171.y.54.o; B.172.y.54.o; B.173.y.54.o; B.174.y.54.o; B.175.y.54.o;
B.176.y.54.o;
B.188.y.54.o; B.189.y.54.o; B.190.y.54.o; B.196.y.54.o; B.202.y.54.o;
B.205.y.54.o;
- 89-
CA 02268756 1999-04-15
WO 98/17647 PCTIUS97/18193
B.206.y.54.o; B.207.y.54.o; B.208.y.54.o; B.209.y.54.o; B.210.y.54.o;
B.211.y.54.o;
B.212.y.54.o; B.213.y.54.o; B.700.y.54.o; B.701.y.54.o; B.702.y.54.o;
B.703.y.54.o;
B.704.y.54.o; B.705.y.54.o; B.706.y.54.o; B.707.y.54.o; B.708.y.54.o;
B.709.y.54.o;
B.710.y.54.o; B.711.y.54.o; B.712.y.54.o; B.713.y.54.o; B.714.y.54.o;
B.715.y.54.o;
B.716.y.54.o; B.717.y.54.o; B.718.y.54.o; B.719.y.54.o; B.720.y.54.o;
B.721.y.54.o;
B.722.y.54.o; B.723.y.54.o; B.724.y.54.o; B.172.z.54.i; B.173.z.54.i;
B.174.z.54.i;
B.175.z.54.i; B.176.z.54.i; B.188.z.54.i; B.189.z.54.i; B.190.z.54.i;
B.196.z.54.i;
B.202.z.54.i; B.205.z.54.i; B.206.z.54.i; B.207.z.54.i; B.208.z.54.i;
B.209.z.54.i;
B.210.z.54.i; B.211.z.54.i; B.212.z.54.i; B.213.z.54.i; B.700.z.54.i;
B.701.z.54.i;
B.702.z.54.i; B.703.z.54.i; B.704.z.54.i; B.705.z.54.i; B.706.z.54.i;
B.707.z.54.i;
B.708.z.54.i; B.709.z.54.i; B.710.z.54.i; B.711.z.54.i; B.712.z.54.i;
B.713.z.54.i;
B.714.z.54.i; B.715.z.54.i; B.716.z.54.i; B.717.z.54.i; B.718.z.54.i;
B.719.z.54.i;
B.720.z.54.i; B.721.z.54.i; B.722.z.54.i; B.723.z.54.i; B.724.z.54.i;
B.3.z.54.o; B.4.z.54.o;
B.7.z.54.o; B.9.z.54.o; B.103.z.54.o; B.106.z.54.o; B.107.z.54.o;
B.108.z.54.o;
B.111.z.54.o; B.114.z.54.o; B.117.z.54.o; B.118.z.54.o; B.119.z.54.o;
B.120.z.54.o;
B.121.z.54.o; B.137.z.54.o; B.138.z.54.o; B.139.z.54.o; B.140.z.54.o;
B.141.z.54.o;
B.142.z.54.o; B.145.z.54.o; B.146.z.54.o; B.147.z.54.o; B.148.z.54.o;
B.149.z.54.o;
B.150.z.54.o; B.151.z.54.o; B.165.z.54.o; B.166.z.54.o; B.167.z.54.o;
B.168.z.54.o;
B.169.z.54.o; B.170.z.54.o; B.171.z.54.o; B.172.z.54.o; B.173.z.54.o;
B.174.z.54.o;
B.175.z.54.o; B.176.z.54.o; B.188.z.54.o; B.189.z.54.o; B.190.z.54.o;
B.196.z.54.o;
B.202.z.54.o; B.205.z.54.o; B.206.z.54.o; B.207.z.54.o; B.208.z.54.o;
B.209.z.54.o;
B.210.z.54.o; B.211.z.54.o; B.212.z.54.o; B.213.z.54.o; B.700.z.54.o;
B.701.z.54.o;
B.702.z.54.o; B.703.z.54.o; B.704.z.54.o; B.705.z.54.o; B.706.z.54.o;
B.707.z.54.o;
B.708.z.54.o; B.709.z.54.o; B.710.z.54.o; B.711.z.54.o; B.712.z.54.o;
B.713.z.54.o;
B.714.z.54.o; B.715.z.54.o; B.716.z.54.o; B.717.z.54.o; B.718.z.54.o;
B.719.z.54.o;
B.720.z.54.o; B.721.z.54.o; B.722.z.54.o; B.723.z.54.o; B.724.z.54.o;
B.172.A.54.i;
B.173.A.54.i; B.174.A.54.i; B.175.A.54.i; B.176.A.54.i; B.188.A.54.i;
B.189.A.54.i;
B.190.A.54.i; B.196.A.54.i; B.202.A.54.i; B.205.A.54.i; B.206.A.54.i;
B.207.A.54.i;
B.208.A.54.i; B.209.A.54.i; B.210.A.54.i; B.211.A.54.i; B.212.A.54.i;
B.213.A.54.i;
B.700.A.54.i; B.701.A.54.i; B.702.A.54.i; B.703.A.54.i; B.704.A.54.i;
B.705.A.54.i;
B.706.A.54.i; B.707.A.54.i; B.708.A.54.i; B.709.A.54.i; B.710.A.54.i;
B.711.A.54.i;
B.712.A.54.i; B.713.A.54.i; B.714.A.54.i; B.715.A.54.i; B.716.A.54.i;
B.717.A.54.i;
B.718.A.54.i; B.719.A.54.i; B.720.A.54.i; B.721.A.54.i; B.722.A.54.i;
B.723.A.54.i;
B.724.A.54.i; B.3.A.54.o; B.4.A.54.o; B.7.A.54.o; B.9.A.54.o; B.103.A.54.o;
B.106.A.54.o;
B.107.A.54.o; B.108.A.54.o; B.111.A.54.o; B.114.A.54.o; B.117.A.54.o;
B.118.A.54.o;
B.119.A.54.o; B.120.A.54.o; B.121.A.54.o; B.137.A.54.o; B.138.A.54.o;
B.139.A.54.o;
B.140.A.54.o; B.141.A.54.o; B.142.A.54.o; B.145.A.54.o; B.146.A.54.o;
B.147.A.54.o;
B.148.A.54.o; B.149.A.54.o; B.150.A.54.o; B.151.A.54.o; B.165.A.54.o;
B.166.A.54.o;
B.167.A.54.o; B.168.A.54.o; B.169.A.54.o; B.170.A.54.o; B.171.A.54.o;
B.172.A.54.o;
B.173.A.54.o; B.174.A.54.o; B.175.A.54.o; B.176.A.54.o; B.188.A.54.o;
B.189.A.54.o;
B.190.A.54.o; B.196.A.54.o; B.202.A.54.o; B.205.A.54.o; B.206.A.54.o;
B.207.A.54.o;
B.208.A.54.o; B.209.A.54.o; B.210.A.54.o; B.211.A.54.o; B.212.A.54.o;
B.213.A.54.o;
B.700.A.54.o; B.701.A.54.o; B.702.A.54.o; B.703.A.54.o; B.704.A.54.o;
B.705.A.54.o;
B.706.A.54.o; B.707.A.54.o; B.708.A.54.o; B.709.A.54.o; B.710.A.54.o;
B.711.A.54.o;
B.712.A.54.o; B.713.A.54.o; B.714.A.54.o; B.715.A.54.o; B.716.A.54.o;
B.717.A.54.o;
B.718.A.54.o; B.719.A.54.o; B.720.A.54.o; B.721.A.54.o; B.722.A.54.o;
B.723.A.54.o;
B.724.A.54.o; B.172.B.54.i; B.173.B.54.i; B.174.B.54.i; B.175.B.54.i;
B.176.B.54.i;
B.188.B.54.i; B.189.B.54.i; B.190.B.54.i; B.196.B.54.i; B.202.B.54.i;
B.205.B.54.i;
B.206.B.54.i; B.207.B.54.i; B.208.B.54.i; B.209.B.54.i; B.210.B.54.i;
B.211.B.54.i;
B.212.B.54.i; B.213.B.54.i; B.700.B.54.i; B.701.B.54.i; B.702.B.54.i;
B.703.B.54.i;
B.704.B.54.i; B.705.B.54.i; B.706.B.54.i; B.707.B.54.i; B.708.B.54.i;
B.709.B.54.i;
B.710.B.54.i; B.711.B.54.i; B.712.B.54.i; B.713.B.54.i; B.714.B.54.i;
B.715.B.54.i;
B.716.B.54.i; B.717.B.54.i; B.718.B.54.i; B.719.B.54.i; B.720.B.54.i;
B.721.B.54.i;
B.722.B.54.i; B.723.B.54.i; B.724.B.54.i; B.3.B.54.o; B.4.B.54.o; B.7.B.54.o;
B.9.B.54.o;
B.103.B.54.o; B.106.B.54.o; B.107.B.54.o; B.108.B.54.o; B.111.B.54.o;
B.114.B.54.o;
B.117.B.54.o; B.118.B.54.o; B.119.B.54.o; B.120.B.54.o; B.121.B.54.o;
B.137.B.54.o;
- 90-
CA 02268756 1999-04-15
WO 98/17647 PCTIUS97/18193
B.138.B.54.o; B.139.B.54.o; B.140.B.54.o; B.141.B.54.o; B.142.B.54.o;
B.145.B.54.o;
B.146.B.54.o; B.147.B.54.o; B.148.B.54.o; B.149.B.54.o; B.150.B.54.o;
B.151.B.54.o;
B.165.B.54.o; B.166.B.54.o; B.167.B.54.o; B.168.B.54.o; B.169.B.54.o;
B.170.B.54.o;
B.171.B.54.o; B.172.B.54.o; B.173.B.54.o; B.174.B.54.o; B.175.B.54.o;
B.176.B.54.o;
B.188.B.54.o; B.189.B.54.o; B.190.B.54.o; B.196.B.54.o; B.202.B.54.o;
B.205.B.54.o;
B.206.B.54.o; B.207.B.54.o; B.208.B.54.o; B.209.B.54.o; B.210.B.54.o;
B.211.B.54.o;
B.212.B.54.o; B.213.B.54.o; B.700.B.54.o; B.701.B.54.o; B.702.B.54.o;
B.703.B.54.o;
B.704.B.54.o; B.705.B.54.o; B.706.B.54.o; B.707.B.54.o; B.708.B.54.o;
B.709.B.54.o;
. B.710.B.54.o; B.711.B.54.o; B.712.B.54.o; B.713.B.54.o; B.714.B.54.o;
B.715.B.54.o;
B.716.B.54.o; B.717.B.54.o; B.718.B.54.o; B.719.B.54.o; B.720.B.54.o;
B.721.B.54.o;
B.722.B.54.o; B.723.B.54.o; B.724.B.54.o C.3.a.54.i; C.4.a.54.i; C.7.a.54.i;
C.9.a.54.i;
C.103.a.54.i; C.106.a.54.i; C.107.a.54.i; C.108.a.54.i; C.111.a.54.i;
C.114.a.54.i;
C.117.a.54.i; C.118.a.54.i; C.119.a.54.i; C.120.a.54.i; C.121.a.54.i;
C.137.a.54.i;
C.138.a.54.i; C.139.a.54.i; C.140.a.54.i; C.141.a.54.i; C.142.a.54.i;
C.145.a.54.i;
C.146.a.54.i; C.147.a.54.i; C.148.a.54.i; C.149.a.54.i; C.150.a.54.i;
C.151.a.54.i;
C.165.a.54.i; C.166.a.54.i; C.167.a.54.i; C.168.a.54.i; C.169.a.54.i;
C.170.a.54.i;
C.171.a.54.i; C.172.a.54.i; C.173.a.54.i; C.174.a.54.i; C.175.a.54.i;
C.176.a.54.i;
C.188.a.54.i; C.189.a.54.i; C.190.a.54.i; C.196.a.54.i; C.202.a.54.i;
C.205.a.54.i;
C.206.a.54.i; C.207.a.54.i; C.208.a.54.i; C.209.a.54.i; C.210.a.54.i;
C.211.a.54.i;
C.212.a.54.i; C.213.a.54.i; C.700.a.54.i; C.701.a.54.i; C.702.a.54.i;
C.703.a.54.i;
C.704.a.54.i; C.705.a.54.i; C.706.a.54.i; C.707.a.54.i; C.708.a.54.i;
C.709.a.54.i;
C.710.a.54.i; C.711.a.54.i; C.712.a.54.i; C.713.a.54.i; C.714.a.54.i;
C.715.a.54.i;
C.716.a.54.i; C.717.a.54.i; C.718.a.54.i; C.719.a.54.i; C.720.a.54.i;
C.721.a.54.i;
C.722.a.54.i; C.723.a.54.i; C.724.a.54.i; C.3.a.54.o; C.4.a.54.o; C.7.a.54.o;
C.9.a.54.o;
C.103.a.54.o; C.106.a.54.o; C.107.a.54.o; C.108.a.54.o; C.111.a.54.o;
C.114.a.54.o;
C.117.a.54.o; C.118.a.54.o; C.119.a.54.o; C.120.a.54.o; C.121.a.54.o;
C.137.a.54.o;
C.138.a.54.o; C.139.a.54.o; C.140.a.54.o; C.141.a.54.o; C.142.a.54.o;
C.145.a.54.o;
C.146.a.54.o; C.147.a.54.o; C.148.a.54.o; C.149.a.54.o; C.150.a.54.o;
C.151.a.54.o;
C.165.a.54.o; C.166.a.54.o; C.167.a.54.o; C.168.a.54.o; C.169.a.54.o;
C.170.a.54.o;
C.171.a.54.o; C.172.a.54.o; C.173.a.54.o; C.174.a.54.o; C.175.a.54.o;
C.176.a.54.o;
C.188.a.54.o; C.189.a.54.o; C.190.a.54.o; C.196.a.54.o; C.202.a.54.o;
C.205.a.54.o;
C.206.a.54.o; C.207.a.54.o; C.208.a.54.o; C.209.a.54.o; C.210.a.54.o;
C.211.a.54.o;
C.212.a.54.o; C.213.a.54.o; C.700.a.54.o; C.701.a.54.o; C.702.a.54.o;
C.703.a.54.o;
C.704.a.54.o; C.705.a.54.o; C.706.a.54.o; C.707.a.54.o; C.708.a.54.o;
C.709.a.54.o;
C.710.a.54.o; C.711.a.54.o; C.712.a.54.o; C.713.a.54.o; C.714.a.54.o;
C.715.a.54.o;
C:716.a.54.o; C.717.a.54.o; C.718.a.54.o; C.719.a.54.o; C.720.a.54.o;
C.721.a.54.o;
C.722.a.54.o; C.723.a.54.o; C.724.a.54.o; C.172.b.54.i; C.173.b.54.i;
C.174.b.54.i;
C.175.b.54.i; C.176.b.54.i; C.188.b.54.i; C.189.b.54.i; C.190.b.54.i;
C.196.b.54.i;
C.202.b.54.i; C.205.b.54.i; C.206.b.54.i; C.207.b.54.i; C.208.b.54.i;
C.209.b.54.i;
C.210.b.54.i; C.211.b.54.i; C.212.b.54.i; C.213.b.54.i; C.700.b.54.i;
C.701.b.54.i;
C.702.b.54.i; C.703.b.54.i; C.704.b.54.i; C.705.b.54.i; C.706.b.54.i;
C.707.b.54.i;
C.708.b.54.i; C.709.b.54.i; C.710.b.54.i; C.711.b.54.i; C.712.b.54.i;
C.713.b.54.i;
C.714.b.54.i; C.715.b.54.i; C.716.b.54.i; C.717.b.54.i; C.718.b.54.i;
C.719.b.54.i;
C.720.b.54.i; C.721.b.54.i; C.722.b.54.i; C.723.b.54.i; C.724.b.54.i;
C.3.b.54.o; C.4.b.54.o;
C.7.b.54.o; C.9.b.54.o; C.103.b.54.o; C.106.b.54.o; C.107.b.54.o;
C.108.b.54.o;
C.111.b.54.o; C.114.b.54.o; C.117.b.54.o; C.118.b.54.o; C.119.b.54.o;
C.120.b.54.o;
C.121.b.54.o; C.137.b.54.o; C.138.b.54.o; C.139.b.54.o; C.140.b.54.o;
C.141.b.54.o;
C.142.b.54.o; C.145.b.54.o; C.146.b.54.o; C.147.b.54.o; C.148.b.54.o;
C.149.b.54.o;
C.150.b.54.o; C.151.b.54.o; C.165.b.54.o; C.166.b.54.o; C.167.b.54.o;
C.168.b.54.o;
C.169.b.54.o; C.170.b.54.o; C.171.b.54.o; C.172.b.54.o; C.173.b.54.o;
C.174.b.54.o;
C.175.b.54.o; C.176.b.54.o; C.188.b.54.o; C.189.b.54.o; C.190.b.54.o;
C.196.b.54.o;
C.202.b.54.o; C.205.b.54.o; C.206.b.54.o; C.207.b.54.o; C.208.b.54.o;
C.209.b.54.o;
C.210.b.54.o; C.211.b.54.o; C.212.b.54.o; C.213.b.54.o; C.700.b.54.o;
C.701.b.54.o;
C.702.b.54.o; C.703.b.54.o; C.704.b.54.o; C.705.b.54.o; C.706.b.54.o;
C.707.b.54.o;
C.708.b.54.o; C.709.b.54.o; C.710.b.54.o; C.711.b.54.o; C.712.b.54.o;
C.713.b.54.o;
C.714.b.54.o; C.715.b.54.o; C.716.b.54.o; C.717.b.54.o; C.718.b.54.o;
C.719.b.54.o;
- 91-
_
CA 02268756 1999-04-15
WO 98/17647 PCT/US97/18193
C.720.b.54.o; C.721.b.54.o; C.722.b.54.o; C.723.b.54.o; C.724.b.54.o;
C.172.x.54.i;
C.173.x.54.i; C.174.x.54.i; C.175.x.54.i; C.176.x.54.i; C.188.x.54.i;
C.189.x.54.i;
C.190.x.54.i; C.196.x.54.i; C.202.x.54.i; C.205.x.54.i; C.206.x.54.i;
C.207.x.54.i;
C.208.x.54.i; C.209.x.54.i; C.210.x.54.i; C.211.x.54.i; C.212.x.54.i;
C.213.x.54.i;
C.700.x.54.i; C.701.x.54.i; C.702.x.54.i; C.703.x.54.i; C.704.x.54.i;
C.705.x.54.i;
C.706.x.54.i; C.707.x.54.i; C.708.x.54.i; C.709.x.54.i; C.710.x.54.i;
C.711.x.54.i;
C.712.x.54.i; C.713.x.54.i; C.714.x.54.i; C.715.x.54.i; C.716.x.54.i;
C.717.x.54.i;
C.718.x.54.i; C.719.x.54.i; C.720.x.54.i; C.721.x.54.i; C.722.x.54.i;
C.723.x.54.i;
C.724.x.54.i; C.3.x.54.o; C.4.x.54.o; C.7.x.54.o; C.9.x.54.o; C.103.x.54.o;
C.106.x.54.o;
C.107.x.54.o; C.108.x.54.o; C.111.x.54.o; C.114.x.54.o; C.117.x.54.o;
C.118.x.54.o;
C.119.x.54.o; C.120.x.54.o; C.121.x.54.o; C.137.x.54.o; C.138.x.54.o;
C.139.x.54.o;
C.140.x.54.o; C.141.x.54.o; C.142.x.54.o; C.145.x.54.o; C.146.x.54.o;
C.147.x.54.o;
C.148.x.54.o; C.149.x.54.o; C.150.x.54.o; C.151.x.54.o; C.165.x.54.o;
C.166.x.54.o;
C.167.x.54.o; C.168.x.54.o; C.169.x.54.o; C.170.x.54.o; C.171.x.54.o;
C.172.x.54.o;
C.173.x.54.o; C.174.x.54.o; C.175.x.54.o; C.176.x.54.o; C.188.x.54.o;
C.189.x.54.o;
C.190.x.54.o; C.196.x.54.o; C.202.x.54.o; C.205.x.54.o; C.206.x.54.o;
C.207.x.54.o;
C.208.x.54.o; C.209.x.54.o; C.210.x.54.o; C.211.x.54.o; C.212.x.54.o;
C.213.x.54.o;
C.700.x.54.o; C.701.x.54.o; C.702.x.54.o; C.703.x.54.o; C.704.x.54.o;
C.705.x.54.o;
C.706.x.54.o; C.707.x.54.o; C.708.x.54.o; C.709.x.54.o; C.710.x.54.o;
C.711.x.54.o;
C.712.x.54.o; C.713.x.54.o; C.714.x.54.o; C.715.x.54.o; C.716.x.54.o;
C.717.x.54.o;
C.718.x.54.o; C.719.x.54.o; C.720.x.54.o; C.721.x.54.o; C.722.x.54.o;
C.723.x.54.o;
C.724.x.54.o; C.172.y.54.i; C.173.y.54.i; C.174.y.54.i; C.175.y.54.i;
C.176.y.54.i;
C.188.y.54.i; C.189.y.54.i; C.190.y.54.i; C.196.y.54.i; C.202.y.54.i;
C.205.y.54.i;
C.206.y.54.i; C.207.y.54.i; C.208.y.54.i; C.209.y.54.i; C.210.y.54.i;
C.211.y.54.i;
C.212.y.54.i; C.213.y.54.i; C.700.y.54.i; C.701.y.54.i; C.702.y.54.i;
C.703.y.54.i;
C.704.y.54.i; C.705.y.54.i; C.706.y.54.i; C.707.y.54.i; C.708.y.54.i;
C.709.y.54.i;
C.710.y.54.i; C.711.y.54.i; C.712.y.54.i; C.713.y.54.i; C.714.y.54.i;
C.715.y.54.i;
C.716.y.54.i; C.717.y.54.i; C.718.y.54.i; C.719.y.54.i; C.720.y.54.i;
C.721.y.54.i;
C.722.y.54.i; C.723.y.54.i; C.724.y.54.i; C.3.y.54.o; C.4.y.54.o; C.7.y.54.o;
C.9.y.54.o;
C.103.y.54.o; C.106.y.54.o; C.107.y.54.o; C.108.y.54.o; C.111.y.54.o;
C.114.y.54.o;
C.117.y.54.o; C.118.y.54.o; C.119.y.54.o; C.120.y.54.o; C.121.y.54.o;
C.137.y.54.o;
C.138.y.54.o; C.139.y.54.o; C.140.y.54.o; C.141.y.54.o; C.142.y.54.o;
C.145.y.54.o;
C.146.y.54.o; C.147.y.54.o; C.148.y.54.o; C.149.y.54.o; C.150.y.54.o;
C.151.y.54.o;
C.165.y.54.o; C.166.y.54.o; C.167.y.54.o; C.168.y.54.o; C.169.y.54.o;
C.170.y.54.o;
C.171.y.54.o; C.172.y.54.o; C.173.y.54.o; C.174.y.54.o; C.175.y.54.o;
C.176.y.54.o;
C.188.y.54.o; C.189.y.54.o; C.190.y.54.o; C.196.y.54.o; C.202.y.54.o;
C.205.y.54.o;
C.206.y.54.o; C.207.y.54.o; C.208.y.54.o; C.209.y.54.o; C.210.y.54.o;
C.211.y.54.o;
C.212.y.54.o; C.213.y.54.o; C.700.y.54.o; C.701.y.54.o; C.702.y.54.o;
C.703.y.54.o;
C.704.y.54.o; C.705.y.54.o; C.706.y.54.o; C.707.y.54.o; C.708.y.54.o;
C.709.y.54.o;
C.710.y.54.o; C.711.y.54.o; C.712.y.54.o; C.713.y.54.o; C.714.y.54.o;
C.715.y.54.o;
C.716.y.54.o; C.717.y.54.o; C.718.y.54.o; C.719.y.54.o; C.720.y.54.o;
C.721.y.54.o;
C.722.y.54.o; C.723.y.54.o; C.724.y.54.o; C.172.z.54.i; C.173.z.54.i;
C.174.z.54.i;
C.175.z.54.i; C.176.z.54.i; C.188.z.54.i; C.189.z.54.i; C.190.z.54.i;
C.196.z.54.i;
C.202.z.54.i; C.205.z.54.i; C.206.z.54.i; C.207.z.54.i; C.208.z.54.i;
C.209.z.54.i;
C.210.z.54.i; C.211.z.54.i; C.212.z.54.i; C.213.z.54.i; C.700.z.54.i;
C.701.z.54.i;
C.702.z.54.i; C.703.z.54.i; C.704.z.54.i; C.705.z.54.i; C.706.z.54.i;
C.707.z.54.i;
C.708.z.54.i; C.709.z.54.i; C.710.z.54.i; C.711.z.54.i; C.712.z.54.i;
C.713.z.54.i;
C.714.z.54.i; C.715.z.54.i; C.716.z.54.i; C.717.z.54.i; C.718.z.54.i;
C.719.z.54.i;
C.720.z.54.i; C.721.z.54.i; C.722.z.54.i; C.723.z.54.i; C.724.z.54.i;
C.3.z.54.o; C.4.z.54.o;
C.7.z.54.o; C.9.z.54.o; C.103.z.54.o; C.106.z.54.o; C.107.z.54.o;
C.108.z.54.o;
C.111.z.54.o; C.114.z.54.o; C.117.z.54.o; C.118.z.54.o; C.119.z.54.o;
C.120.z.54.o;
C.121.z.54.o; C.137.z.54.o; C.138.z.54.o; C.139.z.54.o; C.140.z.54.o;
C.141.z.54.o;
C.142.z.54.o; C.145.z.54.o; C.146.z.54.o; C.147.z.54.o; C.148.z.54.o;
C.149.z.54.o;
C.150.z.54.o; C.151.z.54.o; C.165.z.54.o; C.166.z.54.o; C.167.z.54.o;
C.168.z.54.o;
C.169.z.54.o; C.170.z.54.o; C.171.z.54.o; C.172.z.54.o; C.173.z.54.o;
C.174.z.54.o;
C.175.z.54.o; C.176.z.54.o; C.188.z.54.o; C.189.z.54.o; C.190.z.54.o;
C.196.z.54.o;
- 92-
--__
CA 02268756 1999-04-15
WO 98/17647 PCT/U597/18193
C.202.z.54.o; C.205.z.54.o; C.206.z.54.o; C.207.z.54.o; C.208.z.54.o;
C.209.z.54.o;
C.210.z.54.o; C.211.z.54.o; C.212.z.54.o; C.213.z.54.o; C.700.z.54.o;
C.701.z.54.o;
C.702.z.54.o; C.703.z.54.o; C.704.z.54.o; C.705.z.54.o; C.706.z.54.o;
C.707.z.54.o;
C.708.z.54.o; C.709.z.54.o; C.710.z.54.o; C.711.z.54.o; C.712.z.54.o;
C.713.z.54.o;
C.714.z.54.o; C.715.z.54.o; C.716.z.54.o; C.717.z.54.o; C.718.z.54.o;
C.719.z.54.o;
C.720.z.54.o; C.721.z.54.o; C.722.z.54.o; C.723.z.54.o; C.724.z.54.o;
C.172.A.54.i;
C.173.A.54.i; C.174.A.54.i; C.175.A.54.i; C.176.A.54.i; C.188.A.54.i;
C.189.A.54.i;
C.190.A.54.i; C.196.A.54.i; C.202.A.54.i; C.205.A.54.i; C.206.A.54.i;
C.207.A.54.i;
. C.208.A.54.i; C.209.A.54.i; C.210.A.54.i; C.211.A.54.i; C.212.A.54.i;
C.213.A.54.i;
C.700.A.54.i; C.701.A.54.i; C.702.A.54.i; C.703.A.54.i; C.704.A.54.i;
C.705.A.54.i;
C.706.A.54.i; C.707.A.54.i; C.708.A.54.i; C.709.A.54.i; C.710.A.54.i;
C.711.A.54.i;
C.712.A.54.i; C.713.A.54.i; C.714.A.54.i; C.715.A.54.i; C.716.A.54.i;
C.717.A.54.i;
C.718.A.54.i; C.719.A.54.i; C.720.A.54.i; C.721.A.54.i; C.722.A.54.i;
C.723.A.54.i;
C.724.A.54.i; C.3.A.54.o; C.4.A.54.o; C.7.A.54.o; C.9.A.54.o; C.103.A.54.o;
C.106.A.54.o; C.107.A.54.o; C.108.A.54.o; C.111.A.54.o; C.114.A.54.o;
C.117.A.54.o;
C.118.A.54.o; C.119.A.54.o; C.120.A.54.o; C.121.A.54.o; C.137.A.54.o;
C.138.A.54.o;
C.139.A.54.o; C.140.A.54.o; C.141.A.54.o; C.142.A.54.o; C.145.A.54.o;
C.146.A.54.o;
C.147.A.54.o; C.148.A.54.o; C.149.A.54.o; C.150.A.54.o; C.151.A.54.o;
C.165.A.54.o;
C.166.A.54.o; C.167.A.54.o; C.168.A.54.o; C.169.A.54.o; C.170.A.54.o;
C.171.A.54.o;
C.172.A.54.o; C.173.A.54.o; C.174.A.54.o; C.175.A.54.o; C.176.A.54.o;
C.188.A.54.o;
C.189.A.54.o; C.190.A.54.o; C.196.A.54.o; C.202.A.54.o; C.205.A.54.o;
C.206.A.54.o;
C.207.A.54.o; C.208.A.54.o; C.209.A.54.o; C.210.A.54.o; C.211.A.54.o;
C.212.A.54.o;
C.213.A.54.o; C.700.A.54.o; C.701.A.54.o; C.702.A.54.o; C.703.A.54.o;
C.704.A.54.o;
C.705.A.54.o; C.706.A.54.o; C.707.A.54.o; C.708.A.54.o; C.709.A.54.o;
C.710.A.54.o;
C.711.A.54.o; C.712.A.54.o; C.713.A.54.o; C.714.A.54.o; C.715.A.54.o;
C.716.A.54.o;
C.717.A.54.o; C.718.A.54.o; C.719.A.54.o; C.720.A.54.o; C.721.A.54.o;
C.722.A.54.o;
C.723.A.54.o; C.724.A.54.o; C.172.B.54.i; C.173.B.54.i; C.174.B.54.i;
C.175.B.54.i;
C.176.B.54.i; C.188.B.54.i; C.189.B.54.i; C.190.B.54.i; C.196.B.54.i;
C.202.B.54.i;
C.205.B.54.i; C.206.B.54.i; C.207.B.54.i; C.208.B.54.i; C.209.B.54.i;
C.210.B.54.i;
C.211.B.54.i; C.212.B.54.i; C.213.B.54.i; C.700.B.54.i; C.701.B.54.i;
C.702.B.54.i;
C.703.B.54.i; C.704.B.54.i; C.705.B.54.i; C.706.B.54.i; C.707.B.54.i;
C.708.B.54.i;
C.709.B.54.i; C.710.B.54.i; C.711.B.54.i; C.712.B.54.i; C.713.B.54.i;
C.714.B.54.i;
C.715.B.54.i; C.716.B.54.i; C.717.B.54.i; C.718.B.54.i; C.719.B.54.i;
C.720.B.54.i;
C.721.B.54.i; C.722.B.54.i; C.723.B.54.i; C.724.B.54.i; C.3.B.54.o;
C.4.B.54.o; C.7.B.54.o;
C.9.B.54.o; C.103.B.54.o; C.106.B.54.o; C.107.B.54.o; C.108.B.54.o;
C.111.B.54.o;
C.114.B.54.o; C.117.B.54.o; C.118.B.54.o; C.119.B.54.o; C.120.B.54.o;
C.121.B.54.o;
C.137.B.54.o; C.138.B.54.o; C.139.B.54.o; C.140.B.54.o; C.141.B.54.o;
C.142.B.54.o;
C.145.B.54.o; C.146.B.54.o; C.147.B.54.o; C.148.B.54.o; C.149.B.54.o;
C.150.B.54.o;
C.151.B.54.o; C.165.B.54.o; C.166.B.54.o; C.167.B.54.o; C.168.B.54.o;
C.169.B.54.o;
C.170.B.54.o; C.171.B.54.o; C.172.B.54.o; C.173.B.54.o; C.174.B.54.o;
C.175.B.54.o;
C.176.B.54.o; C.188.B.54.o; C.189.B.54.o; C.190.B.54.o; C.196.B.54.o;
C.202.B.54.o;
C.205.B.54.o; C.206.B.54.o; C.207.B.54.o; C.208.B.54.o; C.209.B.54.o;
C.210.B.54.o;
C.211.B.54.o; C.212.B.54.o; C.213.B.54.o; C.700.B.54.o; C.701.B.54.o;
C.702.B.54.o;
C.703.B.54.o; C.704.B.54.o; C.705.B.54.o; C.706.B.54.o; C.707.B.54.o;
C.708.B.54.o;
C.709.B.54.o; C.710.B.54.o; C.711.B.54.o; C.712.B.54.o; C.713.B.54.o;
C.714.B.54.o;
C.715.B.54.o; C.716.B.54.o; C.717.B.54.o; C.718.B.54.o; C.719.B.54.o;
C.720.B.54.o;
C.721.B.54.o; C.722.B.54.o; C.723.B.54.o; C.724.B.54.o; D.3.a.4.i; D.4.a.4.i;
D.7.a.4.i;
D.9.a.4.i; D.103.a.4.i; D.106.a.4.i; D.107.a.4.i; D.108.a.4.i; D.111.a.4.i;
D.114.a.4.i;
D.117.a.4.i; D.118.a.4.i; D.119.a.4.i; D.120.a.4.i; D.121.a.4.i; D.137.a.4.i;
D.138.a.4.i;
D.139.a.4.i; D.140.a.4.i; D.141.a.4.i; D.142.a.4.i; D.145.a.4.i; D.146.a.4.i;
D.147.a.4.i;
D.148.a.4.i; D.149.a.4.i; D.150.a.4.i; D.151.a.4.i; D.165.a.4.i; D.166.a.4.i;
D.167.a.4.i;
D.168.a.4.i; D.169.a.4.i; D.170.a.4.i; D.171.a.4.i; D.172.a.4.i; D.173.a.4.i;
D.174.a.4.i;
D.175.a.4.i; D.176.a.4.i; D.188.a.4.i; D.189.a.4.i; D.190.a.4.i; D.196.a.4.i;
D.202.a.4.i;
D.205.a.4.i; D.206.a.4.i; D.207.a.4.i; D.208.a.4.i; D.209.a.4.i; D.210.a.4.i;
D.211.a.4.i;
D.212.a.4.i; D.213.a.4.i; D.700.a.4.i; D.701.a.4.i; D.702.a.4.i; D.703.a.4.i;
D.704.a.4.i;
- 93-
_
CA 02268756 1999-04-15
WO 98/17647 PCT/US97/18193
D.705.a.4.i; D.706.a.4.i; D.707.a.4.i; D.708.a.4.i; D.709.a.4.i; D.710.a.4.i;
D.711.a.4.i;
D.712.a.4.i; D.713.a.4.i; D.714.a.4.i; D.715.a.4.i; D.716.a.4.i; D.717.a.4.i;
D.718.a.4.i;
D.719.a.4.i; D.720.a.4.i; D.721.a.4.i; D.722.a.4.i; D.723.a.4.i; D.724.a.4.i;
D.3.a.4.o;
D.4.a.4.o; D.7.a.4.o; D.9.a.4.o; D.103.a.4.o; D.106.a.4.o; D.107.a.4.o;
D.108.a.4.o;
D.111.a.4.o; D.114.a.4.o; D.117.a.4.o; D.118.a.4.o; D.119.a.4.o; D.120.a.4.o;
D.121.a.4.o; D.137.a.4.o; D.138.a.4.o; D.139.a.4.o; D.140.a.4.o; D.141.a.4.o;
D.142.a.4.o; D.145.a.4.o; D.146.a.4.o; D.147.a.4.o; D.148.a.4.o; D.149.a.4.o;
D.150.a.4.o; D.151.a.4.o; D.165.a.4.o; D.166.a.4.o; D.167.a.4.o; D.168.a.4.o;
D.169.a.4.o; D.170.a.4.o; D.171.a.4.o; D.172.a.4.o; D.173.a.4.o; D.174.a.4.o;
D.175.a.4.o; D.176.a.4.o; D.188.a.4.o; D.189.a.4.o; D.190.a.4.o; D.196.a.4.o;
D.202.a.4.o; D.205.a.4.o; D.206.a.4.o; D.207.a.4.o; D.208.a.4.o; D.209.a.4.o;
D.210.a.4.o; D.211.a.4.o; D.212.a.4.o; D.213.a.4.o; D.700.a.4.o; D.701.a.4.o;
D.702.a.4.o; D.703.a.4.o; D.704.a.4.o; D.705.a.4.o; D.706.a.4.o; D.707.a.4.o;
D.708.a.4.o; D.709.a.4.o; D.710.a.4.o; D.711.a.4.o; D.712.a.4.o; D.713.a.4.o;
D.714.a.4.o; D.715.a.4.o; D.716.a.4.o; D.717.a.4.o; D.718.a.4.o; D.719.a.4.o;
D.720.a.4.o; D.721.a.4.o; D.722.a.4.o; D.723.a.4.o; D.724.a.4.o; D.172.b.4.i;
D.173.b.4.i;
D.174.b.4.i; D.175.b.4.i; D.176.b.4.i; D.188.b.4.i; D.189.b.4.i; D.190.b.4.i;
D.196.b.4.i;
D.202.b.4.i; D.205.b.4.i; D.206.b.4.i; D.207.b.4.i; D.208.b.4.i; D.209.b.4.i;
D.210.b.4.i;
D.211.b.4.i; D.212.b.4.i; D.213.b.4.i; D.700.b.4.i; D.701.b.4.i; D.702.b.4.i;
D.703.b.4.i;
D.704.b.4.i; D.705.b.4.i; D.706.b.4.i; D.707.b.4.i; D.708.b.4.i; D.709.b.4.i;
D.710.b.4.i;
D.711.b.4.i; D.712.b.4.i; D.713.b.4.i; D.714.b.4.i; D.715.b.4.i; D.716.b.4.i;
D.717.b.4.i;
D.718.b.4.i; D.719.b.4.i; D.720.b.4.i; D.721.b.4.i; D.722.b.4.i; D.723.b.4.i;
D.724.b.4.i;
D.3.b.4.o; D.4.b.4.o; D.7.b.4.o; D.9.b.4.o; D.103.b.4.o; D.106.b.4.o;
D.107.b.4.o;
D.108.b.4.o; D.111.b.4.o; D.114.b.4.o; D.117.b.4.o; D.118.b.4.o; D.119.b.4.o;
D.120.b.4.o; D.121.b.4.o; D.137.b.4.o; D.138.b.4.o; D.139.b.4.o; D.140.b.4.o;
D.141.b.4.o; D.142.b.4.o; D.145.b.4.o; D.146.b.4.o; D.147.b.4.o; D.148.b.4.o;
D.149.b.4.o; D.150.b.4.o; D.151.b.4.o; D.165.b.4.o; D.166.b.4.o; D.167.b.4.o;
D.168.b.4.o; D.169.b.4.o; D.170.b.4.o; D.171.b.4.o; D.172.b.4.o; D.173.b.4.o;
D.174.b.4.o; D.175.b.4.o; D.176.b.4.o; D.188.b.4.o; D.189.b.4.o; D.190.b.4.o;
D.196.b.4.o; D.202.b.4.o; D.205.b.4.o; D.206.b.4.o; D.207.b.4.o; D.208.b.4.o;
D.209.b.4.o; D.210.b.4.o; D.211.b.4.o; D.212.b.4.o; D.213.b.4.o; D.700.b.4.o;
D.701.b.4.o; D.702.b.4.o; D.703.b.4.o; D.704.b.4.o; D.705.b.4.o; D.706.b.4.o;
D.707.b.4.o; D.708.b.4.o; D.709.b.4.o; D.710.b.4.o; D.711.b.4.o; D.712.b.4.o;
D.713.b.4.o; D.714.b.4.o; D.715.b.4.o; D.716.b.4.o; D.717.b.4.o; D.718.b.4.o;
D.719.b.4.o; D.720.b.4.o; D.721.b.4.o; D.722.b.4.o; D.723.b.4.o; D.724.b.4.o;
D.172.x.4.i;
D.173.x.4.i; D.174.x.4.i; D.175.x.4.i; D.176.x.4.i; D.188.x.4.i; D.189.x.4.i;
D.190.x.4.i;
D.196.x.4.i; D.202.x.4.i; D.205.x.4.i; D.206.x.4.i; D.207.x.4.i; D.208.x.4.i;
D.209.x.4.i;
D.210.x.4.i; D.211.x.4.i; D.212.x.4.i; D.213.x.4.i; D.700.x.4.i; D.701.x.4.i;
D.702.x.4.i;
D.703.x.4.i; D.704.x.4.i; D.705.x.4.i; D.706.x.4.i; D.707.x.4.i; D.708.x.4.i;
D.709.x.4.i;
D.710.x.4.i; D.711.x.4.i; D.712.x.4.i; D.713.x.4.i; D.714.x.4.i; D.715.x.4.i;
D.716.x.4.i;
D.717.x.4.i; D.718.x.4.i; D.719.x.4.i; D.720.x.4.i; D.721.x.4.i; D.722.x.4.i;
D.723.x.4.i;
D.724.x.4.i; D.3.x.4.o; D.4.x.4.o; D.7.x.4.o; D.9.x.4.o; D.103.x.4.o;
D.106.x.4.o;
D.107.x.4.o; D.108.x.4.o; D.111.x.4.o; D.114.x.4.o; D.117.x.4.o; D.118.x.4.o;
D.119.x.4.o; D.120.x.4.o; D.121.x.4.o; D.137.x.4.o; D.138.x.4.o; D.139.x.4.o;
D.140.x.4.o; D.141.x.4.o; D.142.x.4.o; D.145.x.4.o; D.146.x.4.o; D.147.x.4.o;
D.148.x.4.o; D.149.x.4.o; D.150.x.4.o; D.151.x.4.o; D.165.x.4.o; D.166.x.4.o;
D.167.x.4.o; D.168.x.4.o; D.169.x.4.o; D.170.x.4.o; D.171.x.4.o; D.172.x.4.o;
D.173.x.4.o; D.174.x.4.o; D.175.x.4.o; D.176.x.4.o; D.188.x.4.o; D.189.x.4.o;
D.190.x.4.o; D.196.x.4.o; D.202.x.4.o; D.205.x.4.o; D.206.x.4.o; D.207.x.4.o;
D.208.x.4.o; D.209.x.4.o; D.210.x.4.o; D.211.x.4.o; D.212.x.4.o; D.213.x.4.o;
D.700.x.4.o; D.701.x.4.o; D.702.x.4.o; D.703.x.4.o; D.704.x.4.o; D.705.x.4.o;
D.706.x.4.o; D.707.x.4.o; D.708.x.4.o; D.709.x.4.o; D.710.x.4.o; D.711.x.4.o;
D.712.x.4.o; D.713.x.4.o; D.714.x.4.o; D.715.x.4.o; D.716.x.4.o; D.717.x.4.o;
D.718.x.4.o; D.719.x.4.o; D.720.x.4.o; D.721.x.4.o; D.722.x.4.o; D.723.x.4.o;
D.724.x.4.o; D.172.y.4.i; D.173.y.4.i; D.174.y.4.i; D.175.y.4.i; D.176.y.4.i;
D.188.y.4.i;
D.189.y.4.i; D.190.y.4.i; D.196.y.4.i; D.202.y.4.i; D.205.y.4.i; D.206.y.4.i;
D.207.y.4.i;
- 94-
__
CA 02268756 1999-04-15
WO 98/17647 PCTIUS97/18193
D.208.y.4.i; D.209.y.4.i; D.210.y.4.i; D.211.y.4.i; D.212.y.4.i; D.213.y.4.i;
D.700.y.4.i;
D.701.y.4.i; D.702.y.4.i; D.703.y.4.i; D.704.y.4.i; D.705.y.4.i; D.706.y.4.i;
D.707.y.4.i;
D.708.y.4.i; D.709.y.4.i; D.710.y.4.i; D.711.y.4.i; D.712.y.4.i; D.713.y.4.i;
D.714.y.4.i;
D.715.y.4.i; D.716.y.4.i; D.717.y.4.i; D.718.y.4.i; D.719.y.4.i; D.720.y.4.i;
D.721.y.4.i;
D.722.y.4.i; D.723.y.4.i; D.724.y.4.i; D.3.y.4.o; D.4.y.4.o; D.7.y.4.o;
D.9.y.4.o;
D.103.y.4.o; D.106.y.4.o; D.107.y.4.o; D.108.y.4.o; D.111.y.4.o; D.114.y.4.o;
D.117.y.4.o; D.118.y.4.o; D.119.y.4.o; D.120.y.4.o; D.121.y.4.o; D.137.y.4.o;
D.138.y.4.o; D.139.y.4.o; D.140.y.4.o; D.141.y.4.o; D.142.y.4.o; D.145.y.4.o;
D.146.y.4.o; D.147.y.4.o; D.148.y.4.o; D.149.y.4.o; D.150.y.4.o; D.151.y.4.o;
D.165.y.4.o; D.166.y.4.o; D.167.y.4.o; D.168.y.4.o; D.169.y.4.o; D.170.y.4.o;
D.171.y.4.o; D.172.y.4.o; D.173.y.4.o; D.174.y.4.o; D.175.y.4.o; D.176.y.4.o;
D.188.y.4.o; D.189.y.4.o; D.190.y.4.o; D.196.y.4.o; D.202.y.4.o; D.205.y.4.o;
D.206.y.4.o; D.207.y.4.o; D.208.y.4.o; D.209.y.4.o; D.210.y.4.o; D.211.y.4.o;
D.212.y.4.o; D.213.y.4.o; D.700.y.4.o; D.701.y.4.o; D.702.y.4.o; D.703.y.4.o;
D.704.y.4.o; D.705.y.4.o; D.706.y.4.o; D.707.y.4.o; D.708.y.4.o; D.709.y.4.o;
D.710.y.4.o; D.711.y.4.o; D.712.y.4.o; D.713.y.4.o; D.714.y.4.o; D.715.y.4.o;
D.716.y.4.o; D.717.y.4.o; D.718.y.4.o; D.719.y.4.o; D.720.y.4.o; D.721.y.4.o;
D.722.y.4.o; D.723.y.4.o; D.724.y.4.o; D.172.z.4.i; D.173.z.4.i; D.174.z.4.i;
D.175.z.4.i;
D.176.z.4.i; D.188.z.4.i; D.189.z.4.i; D.190.z.4.i; D.196.z.4.i; D.202.z.4.i;
D.205.z.4.i;
D.206.z.4.i; D.207.z.4.i; D.208.z.4.i; D.209.z.4.i; D.210.z.4.i; D.211.z.4.i;
D.212.z.4.i;
D.213.z.4.i; D.700.z.4.i; D.701.z.4.i; D.702.z.4.i; D.703.z.4.i; D.704.z.4.i;
D.705.z.4.i;
D.706.z.4.i; D.707.z.4.i; D.708.z.4.i; D.709.z.4.i; D.710.z.4.i; D.711.z.4.i;
D.712.z.4.i;
D.713.z.4.i; D.714.z.4.i; D.715.z.4.i; D.716.z.4.i; D.717.z.4.i; D.718.z.4.i;
D.719.z.4.i;
D.720.z.4.i; D.721.z.4.i; D.722.z.4.i; D.723.z.4.i; D.724.z.4.i; D.3.z.4.o;
D.4.z.4.o;
D.7.z.4.o; D.9.z.4.o; D.103.z.4.o; D.106.z.4.o; D.107.z.4.o; D.108.z.4.o;
D.111.z.4.o;
D.114.z.4.o; D.117.z.4.o; D.118.z.4.o; D.119.z.4.o; D.120.z.4.o; D.121.z.4.o;
D.137.z.4.o; D.138.z.4.o; D.139.z.4.o; D.140.z.4.o; D.141.z.4.o; D.142.z.4.o;
D.145.z.4.o; D.146.z.4.o; D.147.z.4.o; D.148.z.4.o; D.149.z.4.o; D.150.z.4.o;
D.151.z.4.o; D.165.z.4.o; D.166.z.4.o; D.167.z.4.o; D.168.z.4.o; D.169.z.4.o;
D.170.z.4.o; D.171.z.4.o; D.172.z.4.o; D.173.z.4.o; D.174.z.4.o; D.175.z.4.o;
D.176.z.4.o; D.188.z.4.o; D.189.z.4.o; D.190.z.4.o; D.196.z.4.o; D.202.z.4.o;
D.205.z.4.o; D.206.z.4.o; D.207.z.4.o; D.208.z.4.o; D.209.z.4.o; D.210.z.4.o;
D.211.z.4.o; D.212.z.4.o; D.213.z.4.o; D.700.z.4.o; D.701.z.4.o; D.702.z.4.o;
D.703.z.4.o; D.704.z.4.o; D.705.z.4.o; D.706.z.4.o; D.707.z.4.o; D.708.z.4.o;
D.709.z.4.o; D.710.z.4.o; D.711.z.4.o; D.712.z.4.o; D.713.z.4.o; D.714.z.4.o;
D.715.z.4.o; D.716.z.4.o; D.717.z.4.o; D.718.z.4.o; D.719.z.4.o; D.720.z.4.o;
D.721.z.4.o; D.722.z.4.o; D.723.z.4.o; D.724.z.4.o; D.172.A.4.i; D.173.A.4.i;
D.174.A.4.i; D.175.A.4.i; D.176.A.4.i; D.188.A.4.i; D.189.A.4.i; D.190.A.4.i;
D.196.A.4.i; D.202.A.4.i; D.205.A.4.i; D.206.A.4.i; D.207.A.4.i; D.208.A.4.i;
D.209.A.4.i; D.210.A.4.i; D.211.A.4.i; D.212.A.4.i; D.213.A.4.i; D.700.A.4.i;
D.701.A.4.i; D.702.A.4.i; D.703.A.4.i; D.704.A.4.i; D.705.A.4.i; D.706.A.4.i;
D.707.A.4.i; D.708.A.4.i; D.709.A.4.i; D.710.A.4.i; D.711.A.4.i; D.712.A.4.i;
D.713.A.4.i; D.714.A.4.i; D.715.A.4.i; D.716.A.4.i; D.717.A.4.i; D.718.A.4.i;
D.719.A.4.i; D.720.A.4.i; D.721.A.4.i; D.722.A.4.i; D.723.A.4.i; D.724.A.4.i;
D.3.A.4.o;
D.4.A.4.o; D.7.A.4.o; D.9.A.4.o; D.103.A.4.o; D.106.A.4.o; D.107.A.4.o;
D.108.A.4.o;
D.111.A.4.o; D.114.A.4.o; D.117.A.4.o; D.118.A.4.o; D.119.A.4.o; D.120.A.4.o;
D.121.A.4.o; D.137.A.4.o; D.138.A.4.o; D.139.A.4.o; D.140.A.4.o; D.141.A.4.o;
D.142.A.4.o; D.145.A.4.o; D.146.A.4.o; D.147.A.4.o; D.148.A.4.o; D.149.A.4.o;
D.150.A.4.o; D.151.A.4.o; D.165.A.4.o; D.166.A.4.o; D.167.A.4.o; D.168.A.4.o;
D.169.A.4.o; D.170.A.4.o; D.171.A.4.o; D.172.A.4.o; D.173.A.4.o; D.174.A.4.o;
D.175.A.4.o; D.176.A.4.o; D.188.A.4.o; D.189.A.4.o; D.190.A.4.o; D.196.A.4.o;
D.202.A.4.o; D.205.A.4.o; D.206.A.4.o; D.207.A.4.o; D.208.A.4.o; D.209.A.4.o;
D.210.A.4.o; D.211.A.4.o; D.212.A.4.o; D.213.A.4.o; D.700.A.4.o; D.701.A.4.o;
D.702.A.4.o; D.703.A.4.o; D.704.A.4.o; D.705.A.4.o; D.706.A.4.o; D.707.A.4.o;
D.708.A.4.o; D.709.A.4.o; D.710.A.4.o; D.711.A.4.o; D.712.A.4.o; D.713.A.4.o;
D.714.A.4.o; D.715.A.4.o; D.716.A.4.o; D.717.A.4.o; D.718.A.4.o; D.719.A.4.o;
- 95-
CA 02268756 1999-04-15
WO 98/17647 PCT/US97/18193
D.720.A.4.o; D.721.A.4.o; D.722.A.4.o; D.723.A.4.o; D.724.A.4.o; D.172.B.4.i;
D.173.B.4.i; D.174.B.4.i; D.175.B.4.i; D.176.B.4.i; D.188.B.4.i; D.189.B.4.i;
D.190.B.4.i;
D.196.B.4.i; D.202.B.4.i; D.205.B.4.i; D.206.B.4.i; D.207.B.4.i; D.208.B.4.i;
D.209.B.4.i;
D.210.B.4.i; D.211.B.4.i; D.212.B.4.i; D.213.B.4.i; D.700.B.4.i; D.701.B.4.i;
D.702.B.4.i;
D.703.B.4.i; D.704.B.4.i; D.705.B.4.i; D.706.B.4.i; D.707.B.4.i; D.708.B.4.i;
D.709.B.4.i;
D.710.B.4.i; D.711.B.4.i; D.712.B.4.i; D.713.B.4.i; D.714.B.4.i; D.715.B.4.i;
D.716.B.4.i;
D.717.B.4.i; D.718.B.4.i; D.719.B.4.i; D.720.B.4.i; D.721.B.4.i; D.722.B.4.i;
D.723.B.4.i;
D.724.B.4.i; D.3.B.4.o; D.4.B.4.o; D.7.B.4.o; D.9.B.4.o; D.103.B.4.o;
D.106.B.4.o;
D.107.B.4.o; D.108.B.4.o; D.111.B.4.o; D.114.B.4.o; D.117.B.4.o; D.118.B.4.o;
D.119.B.4.o; D.120.B.4.o; D.121.B.4.o; D.137.B.4.o; D.138.B.4.o; D.139.B.4.o;
D.140.B.4.o; D.141.B.4.o; D.142.B.4.o; D.145.B.4.o; D.146.B.4.o; D.147.B.4.o;
D.148.B.4.o; D.149.B.4.o; D.150.B.4.o; D.151.B.4.o; D.165.B.4.o; D.166.B.4.o;
D.167.B.4.o; D.168.B.4.o; D.169.B.4.o; D.170.B.4.o; D.171.B.4.o; D.172.B.4.o;
D.173.B.4.o; D.174.B.4.o; D.175.B.4.o; D.176.B.4.o; D.188.B.4.o; D.189.B.4.o;
D.190.B.4.o; D.196.B.4.o; D.202.B.4.o; D.205.B.4.o; D.206.B.4.o; D.207.B.4.o;
D.208.B.4.o; D.209.B.4.o; D.210.B.4.o; D.211.B.4.o; D.212.B.4.o; D.213.B.4.o;
D.700.B.4.o; D.701.B.4.o; D.702.B.4.o; D.703.B.4.o; D.704.B.4.o; D.705.B.4.o;
D.706.B.4.o; D.707.B.4.o; D.708.B.4.o; D.709.B.4.o; D.710.B.4.o; D.711.B.4.o;
D.712.B.4.o; D.713.B.4.o; D.714.B.4.o; D.715.B.4.o; D.716.B.4.o; D.717.B.4.o;
D.718.B.4.o; D.719.B.4.o; D.720.B.4.o; D.721.B.4.o; D.722.B.4.o; D.723.B.4.o;
D.724.B.4.o; E.3.a.54.i; E.4.a.54.i; E.7.a.54.i; E.9.a.54.i; E.103.a.54.i;
E.106.a.54.i;
E.107.a.54.i; E.108.a.54.i; E.111.a.54.i; E.114.a.54.i; E.117.a.54.i;
E.118.a.54.i;
E.119.a.54.i; E.120.a.54.i; E.121.a.54.i; E.137.a.54.i; E.138.a.54.i;
E.139.a.54.i;
E.140.a.54.i; E.141.a.54.i; E.142.a.54.i; E.145.a.54.i; E.146.a.54.i;
E.147.a.54.i;
E.148.a.54.i; E.149.a.54.i; E.150.a.54.i; E.151.a.54.i; E.165.a.54.i;
E.166.a.54.i;
E.167.a.54.i; E.168.a.54.i; E.169.a.54.i; E.170.a.54.i; E.171.a.54.i;
E.172.a.54.i;
E.173.a.54.i; E.174.a.54.i; E.175.a.54.i; E.176.a.54.i; E.188.a.54.i;
E.189.a.54.i;
E.190.a.54.i; E.196.a.54.i; E.202.a.54.i; E.205.a.54.i; E.206.a.54.i;
E.207.a.54.i;
E.208.a.54.i; E.209.a.54.i; E.210.a.54.i; E.211.a.54.i; E.212.a.54.i;
E.213.a.54.i;
E.700.a.54.i; E.701.a.54.i; E.702.a.54.i; E.703.a.54.i; E.704.a.54.i;
E.705.a.54.i;
E.706.a.54.i; E.707.a.54.i; E.708.a.54.i; E.709.a.54.i; E.710.a.54.i;
E.711.a.54.i;
E.712.a.54.i; E.713.a.54.i; E.714.a.54.i; E.715.a.54.i; E.716.a.54.i;
E.717.a.54.i;
E.718.a.54.i; E.719.a.54.i; E.720.a.54.i; E.721.a.54.i; E.722.a.54.i;
E.723.a.54.i;
E.724.a.54.i; E.3.a.54.o; E.4.a.54.o; E.7.a.54.o; E.9.a.54.o; E.103.a.54.o;
E.106.a.54.o;
E.107.a.54.o; E.108.a.54.o; E.111.a.54.o; E.114.a.54.o; E.117.a.54.o;
E.118.a.54.o;
E.119.a.54.o; E.120.a.54.o; E.121.a.54.o; E.137.a.54.o; E.138.a.54.o;
E.139.a.54.o;
E.140.a.54.o; E.141.a.54.o; E.142.a.54.o; E.145.a.54.o; E.146.a.54.o;
E.147.a.54.o;
E.148.a.54.o; E.149.a.54.o; E.150.a.54.o; E.151.a.54.o; E.165.a.54.o;
E.166.a.54.o;
E.167.a.54.o; E.168.a.54.o; E.169.a.54.o; E.170.a.54.o; E.171.a.54.o;
E.172.a.54.o;
E.173.a.54.o; E.174.a.54.o; E.175.a.54.o; E.176.a.54.o; E.188.a.54.o;
E.189.a.54.o;
E.190.a.54.o; E.196.a.54.o; E.202.a.54.o; E.205.a.54.o; E.206.a.54.o;
E.207.a.54.o;
E.208.a.54.o; E.209.a.54.o; E.210.a.54.o; E.211.a.54.o; E.212.a.54.o;
E.213.a.54.o;
E.700.a.54.o; E.701.a.54.o; E.702.a.54.o; E.703.a.54.o; E.704.a.54.o;
E.705.a.54.o;
E.706.a.54.o; E.707.a.54.o; E.708.a.54.o; E.709.a.54.o; E.710.a.54.o;
E.711.a.54.o;
E.712.a.54.o; E.713.a.54.o; E.714.a.54.o; E.715.a.54.o; E.716.a.54.o;
E.717.a.54.o;
E.718.a.54.o; E.719.a.54.o; E.720.a.54.o; E.721.a.54.o; E.722.a.54.o;
E.723.a.54.o;
E.724.a.54.o; E.172.b.54.i; E.173.b.54.i; E.174.b.54.i; E.175.b.54.i;
E.176.b.54.i;
E.188.b.54.i; E.189.b.54.i; E.190.b.54.i; E.196.b.54.i; E.202.b.54.i;
E.205.b.54.i;
E.206.b.54.i; E.207.b.54.i; E.208.b.54.i; E.209.b.54.i; E.210.b.54.i;
E.211.b.54.i;
E.212.b.54.i; E.213.b.54.i; E.700.b.54.i; E.701.b.54.i; E.702.b.54.i;
E.703.b.54.i;
E.704.b.54.i; E.705.b.54.i; E.706.b.54.i; E.707.b.54.i; E.708.b.54.i;
E.709.b.54.i;
E.710.b.54.i; E.711.b.54.i; E.712.b.54.i; E.713.b.54.i; E.714.b.54.i;
E.715.b.54.i;
E.716.b.54.i; E.717.b.54.i; E.718.b.54.i; E.719.b.54.i; E.720.b.54.i;
E.721.b.54.i;
E.722.b.54.i; E.723.b.54.i; E.724.b.54.i; E.3.b.54.o; E.4.b.54.o; E.7.b.54.o;
E.9.b.54.o;
E.103.b.54.o; E.106.b.54.o; E.107.b.54.o; E.108.b.54.o; E.111.b.54.o;
E.114.b.54.o;
E.117.b.54.o; E.118.b.54.o; E.119.b.54.o; E.120.b.54.o; E.121.b.54.o;
E.137.b.54.o;
- 96-
CA 02268756 1999-04-15
WO 98/17647 PCT/US97/18193
E.138.b.54.o; E.139.b.54.o; E.140.b.54.o; E.141.b.54.o; E.142.b.54.o;
E.145.b.54.o;
E.146.b.54.o; E.147.b.54.o; E.148.b.54.o; E.149.b.54.o; E.150.b.54.o;
E.151.b.54.o;
E.165.b.54.o; E.166.b.54.o; E.167.b.54.o; E.168.b.54.o; E.169.b.54.o;
E.170.b.54.o;
E.171.b.54.o; E.172.b.54.o; E.173.b.54.o; E.174.b.54.o; E.175.b.54.o;
E.176.b.54.o;
E.188.b.54.o; E.189.b.54.o; E.190.b.54.o; E.196.b.54.o; E.202.b.54.o;
E.205.b.54.o;
= E.206.b.54.o; E.207.b.54.o; E.208.b.54.o; E.209.b.54.o; E.210.b.54.o;
E.211.b.54.o;
E.212.b.54.o; E.213.b.54.o; E.700.b.54.o; E.701.b.54.o; E.702.b.54.o;
E.703.b.54.o;
E.704.b.54.o; E.705.b.54.o; E.706.b.54.o; E.707.b.54.o; E.708.b.54.o;
E.709.b.54.o;
E.710.b.54.o; E.711.b.54.o; E.712.b.54.o; E.713.b.54.o; E.714.b.54.o;
E.715.b.54.o;
E.716.b.54.o; E.717.b.54.o; E.718.b.54.o; E.719.b.54.o; E.720.b.54.o;
E.721.b.54.o;
E.722.b.54.o; E.723.b.54.o; E.724.b.54.o; E.172.x.54.i; E.173.x.54.i;
E.174.x.54.i;
E.175.x.54.i; E.176.x.54.i; E.188.x.54.i; E.189.x.54.i; E.190.x.54.i;
E.196.x.54.i;
E.202.x.54.i; E.205.x.54.i; E.206.x.54.i; E.207.x.54.i; E.208.x.54.i;
E.209.x.54.i;
E.210.x.54.i; E.211.x.54.i; E.212.x.54.i; E.213.x.54.i; E.700.x.54.i;
E.701.x.54.i;
E.702.x.54.i; E.703.x.54.i; E.704.x.54.i; E.705.x.54.i; E.706.x.54.i;
E.707.x.54.i;
E.708.x.54.i; E.709.x.54.i; E.710.x.54.i; E.711.x.54.i; E.712.x.54.i;
E.713.x.54.i;
E.714.x.54.i; E.715.x.54.i; E.716.x.54.i; E.717.x.54.i; E.718.x.54.i;
E.719.x.54.i;
E.720.x.54.i; E.721.x.54.i; E.722.x.54.i; E.723.x.54.i; E.724.x.54.i;
E.3.x.54.o; E.4.x.54.o;
E.7.x.54.o; E.9.x.54.o; E.103.x.54.o; E.106.x.54.o; E.107.x.54.o;
E.108.x.54.o;
E.111.x.54.o; E.114.x.54.o; E.117.x.54.o; E.118.x.54.o; E.119.x.54.o;
E.120.x.54.o;
E.121.x.54.o; E.137.x.54.o; E.138.x.54.o; E.139.x.54.o; E.140.x.54.o;
E.141.x.54.o;
E.142.x.54.o; E.145.x.54.o; E.146.x.54.o; E.147.x.54.o; E.148.x.54.o;
E.149.x.54.o;
E.150.x.54.o; E.151.x.54.o; E.165.x.54.o; E.166.x.54.o; E.167.x.54.o;
E.168.x.54.o;
E.169.x.54.o; E.170.x.54.o; E.171.x.54.o; E.172.x.54.o; E.173.x.54.o;
E.174.x.54.o;
E.175.x.54.o; E.176.x.54.o; E.188.x.54.o; E.189.x.54.o; E.190.x.54.o;
E.196.x.54.o;
E.202.x.54.o; E.205.x.54.o; E.206.x.54.o; E.207.x.54.o; E.208.x.54.o;
E.209.x.54.o;
E.210.x.54.o; E.211.x.54.o; E.212.x.54.o; E.213.x.54.o; E.700.x.54.o;
E.701.x.54.o;
E.702.x.54.o; E.703.x.54.o; E.704.x.54.o; E.705.x.54.o; E.706.x.54.o;
E.707.x.54.o;
E.708.x.54.o; E.709.x.54.o; E.710.x.54.o; E.711.x.54.o; E.712.x.54.o;
E.713.x.54.o;
E.714.x.54.o; E.715.x.54.o; E.716.x.54.o; E.717.x.54.o; E.718.x.54.o;
E.719.x.54.o;
E.720.x.54.o; E.721.x.54.o; E.722.x.54.o; E.723.x.54.o; E.724.x.54.o;
E.172.y.54.i;
E.173.y.54.i; E.174.y.54.i; E.175.y.54.i; E.176.y.54.i; E.188.y.54.i;
E.189.y.54.i;
E.190.y.54.i; E.196.y.54.i; E.202.y.54.i; E.205.y.54.i; E.206.y.54.i;
E.207.y.54.i;
E.208.y.54.i; E.209.y.54.i; E.210.y.54.i; E.211.y.54.i; E.212.y.54.i;
E.213.y.54.i;
E.700.y.54.i; E.701.y.54.i; E.702.y.54.i; E.703.y.54.i; E.704.y.54.i;
E.705.y.54.i;
E.706.y.54.i; E.707.y.54.i; E.708.y.54.i; E.709.y.54.i; E.710.y.54.i;
E.711.y.54.i;
E.712.y.54.i; E.713.y.54.i; E.714.y.54.i; E.715.y.54.i; E.716.y.54.i;
E.717.y.54.i;
E.718.y.54.i; E.719.y.54.i; E.720.y.54.i; E.721.y.54.i; E.722.y.54.i;
E.723.y.54.i;
E.724.y.54.i; E.3.y.54.o; E.4.y.54.o; E.7.y.54.o; E.9.y.54.o; E.103.y.54.o;
E.106.y.54.o;
E.107.y.54.o; E.108.y.54.o; E.111.y.54.o; E.114.y.54.o; E.117.y.54.o;
E.118.y.54.o;
E.119.y.54.o; E.120.y.54.o; E.121.y.54.o; E.137.y.54.o; E.138.y.54.o;
E.139.y.54.o;
E.140.y.54.o; E.141.y.54.o; E.142.y.54.o; E.145.y.54.o; E.146.y.54.o;
E.147.y.54.o;
E.148.y.54.o; E.149.y.54.o; E.150.y.54.o; E.151.y.54.o; E.165.y.54.o;
E.166.y.54.o;
E.167.y.54.o; E.168.y.54.o; E.169.y.54.o; E.170.y.54.o; E.171.y.54.o;
E.172.y.54.o;
E.173.y.54.o; E.174.y.54.o; E.175.y.54.o; E.176.y.54.o; E.188.y.54.o;
E.189.y.54.o;
E.190.y.54.o; E.196.y.54.o; E.202.y.54.o; E.205.y.54.o; E.206.y.54.o;
E.207.y.54.o;
E.208.y.54.o; E.209.y.54.o; E.210.y.54.o; E.211.y.54.o; E.212.y.54.o;
E.213.y.54.o;
E.700.y.54.o; E.701.y.54.o; E.702.y.54.o; E.703.y.54.o; E.704.y.54.o;
E.705.y.54.o;
E.706.y.54.o; E.707.y.54.o; E.708.y.54.o; E.709.y.54.o; E.710.y.54.o;
E.711.y.54.o;
E.712.y.54.o; E.713.y.54.o; E.714.y.54.o; E.715.y.54.o; E.716.y.54.o;
E.717.y.54.o;
E.718.y.54.o; E.719.y.54.o; E.720.y.54.o; E.721.y.54.o; E.722.y.54.o;
E.723.y.54.o;
E.724.y.54.o; E.172.z.54.i; E.173.z.54.i; E.174.z.54.i; E.175.z.54.i;
E.176.z.54.i;
E.188.z.54.i; E.189.z.54.i; E.190.z.54.i; E.196.z.54.i; E.202.z.54.i;
E.205.z.54.i;
E.206.z.54.i; E.207.z.54.i; E.208.z.54.i; E.209.z.54.i; E.210.z.54.i;
E.211.z.54.i;
E.212.z.54.i; E.213.z.54.i; E.700.z.54.i; E.701.z.54.i; E.702.z.54.i;
E.703.z.54.i;
E.704.z.54.i; E.705.z.54.i; E.706.z.54.i; E.707.z.54.i; E.708.z.54.i;
E.709.z.54.i;
- 97-
CA 02268756 1999-04-15
WO 98/17647 PCT/US97/18193
E.710.z.54.i; E.711.z.54.i; E.712.z.54.i; E.713.z.54.i; E.714.z.54.i;
E.715.z.54.i;
E.716.z.54.i; E.717.z.54.i; E.718.z.54.i; E.719.z.54.i; E.720.z.54.i;
E.721.z.54.i;
E.722.z.54.i; E.723.z.54.i; E.724.z.54.i; E.3.z.54.o; E.4.z.54.o; E.7.z.54.o;
E.9.z.54.o;
E.103.z.54.o; E.106.z.54.o; E.107.z.54.o; E.108.z.54.o; E.111.z.54.o;
E.114.z.54.o;
E.117.z.54.o; E.118.z.54.o; E.119.z.54.o; E.120.z.54.o; E.121.z.54.o;
E.137.z.54.o;
E.138.z.54.o; E.139.z.54.o; E.140.z.54.o; E.141.z.54.o; E.142.z.54.o;
E.145.z.54.o;
E.146.z.54.o; E.147.z.54.o; E.148.z.54.o; E.149.z.54.o; E.150.z.54.o;
E.151.z.54.o;
E.165.z.54.o; E.166.z.54.o; E.167.z.54.o; E.168.z.54.o; E.169.z.54.o;
E.170.z.54.o;
E.171.z.54.o; E.172.z.54.o; E.173.z.54.o; E.174.z.54.o; E.175.z.54.o;
E.176.z.54.o;
E.188.z.54.o; E.189.z.54.o; E.190.z.54.o; E.196.z.54.o; E.202.z.54.o;
E.205.z.54.o;
E.206.z.54.o; E.207.z.54.o; E.208.z.54.o; E.209.z.54.o; E.210.z.54.o;
E.211.z.54.o;
E.212.z.54.o; E.213.z.54.o; E.700.z.54.o; E.701.z.54.o; E.702.z.54.o;
E.703.z.54.o;
E.704.z.54.o; E.705.z.54.o; E.706.z.54.o; E.707.z.54.o; E.708.z.54.o;
E.709.z.54.o;
E.710.z.54.o; E.711.z.54.o; E.712.z.54.o; E.713.z.54.o; E.714.z.54.o;
E.715.z.54.o;
E.716.z.54.o; E.717.z.54.o; E.718.z.54.o; E.719.z.54.o; E.720.z.54.o;
E.721.z.54.o;
E.722.z.54.o; E.723.z.54.o; E.724.z.54.o; E.172.A.54.i; E.173.A.54.i;
E.174.A.54.i;
E.175.A.54.i; E.176.A.54.i; E.188.A.54.i; E.189.A.54.i; E.190.A.54.i;
E.196.A.54.i;
E.202.A.54.i; E.205.A.54.i; E.206.A.54.i; E.207.A.54.i; E.208.A.54.i;
E.209.A.54.i;
E.210.A.54.i; E.211.A.54.i; E.212.A.54.i; E.213.A.54.i; E.700.A.54.i;
E.701.A.54.i;
E.702.A.54.i; E.703.A.54.i; E.704.A.54.i; E.705.A.54.i; E.706.A.54.i;
E.707.A.54.i;
E.708.A.54.i; E.709.A.54.i; E.710.A.54.i; E.711.A.54.i; E.712.A.54.i;
E.713.A.54.i;
E.714.A.54.i; E.715.A.54.i; E.716.A.54.i; E.717.A.54.i; E.718.A.54.i;
E.719.A.54.i;
E.720.A.54.i; E.721.A.54.i; E.722.A.54.i; E.723.A.54.i; E.724.A.54.i;
E.3.A.54.o;
E.4.A.54.o; E.7.A.54.o; E.9.A.54.o; E.103.A.54.o; E.106.A.54.o; E.107.A.54.o;
E.108.A.54.o; E.111.A.54.o; E.114.A.54.o; E.117.A.54.o; E.118.A.54.o;
E.119.A.54.o;
E.120.A.54.o; E.121.A.54.o; E.137.A.54.o; E.138.A.54.o; E.139.A.54.o;
E.140.A.54.o;
E.141.A.54.o; E.142.A.54.o; E.145.A.54.o; E.146.A.54.o; E.147.A.54.o;
E.148.A.54.o;
E.149.A.54.o; E.150.A.54.o; E.151.A.54.o; E.165.A.54.o; E.166.A.54.o;
E.167.A.54.o;
E.168.A.54.o; E.169.A.54.o; E.170.A.54.o; E.171.A.54.o; E.172.A.54.o;
E.173.A.54.o;
E.174.A.54.o; E.175.A.54.o; E.176.A.54.o; E.188.A.54.o; E.189.A.54.o;
E.190.A.54.o;
E.196.A.54.o; E.202.A.54.o; E.205.A.54.o; E.206.A.54.o; E.207.A.54.o;
E.208.A.54.o;
E.209.A.54.o; E.210.A.54.o; E.211.A.54.o; E.212.A.54.o; E.213.A.54.o;
E.700.A.54.o;
E.701.A.54.o; E.702.A.54.o; E.703.A.54.o; E.704.A.54.o; E.705.A.54.o;
E.706.A.54.o;
E.707.A.54.o; E.708.A.54.o; E.709.A.54.o; E.710.A.54.o; E.711.A.54.o;
E.712.A.54.o;
E.713.A.54.o; E.714.A.54.o; E.715.A.54.o; E.716.A.54.o; E.717.A.54.o;
E.718.A.54.o;
E.719.A.54.o; E.720.A.54.o; E.721.A.54.o; E.722.A.54.o; E.723.A.54.o;
E.724.A.54.o;
E.172.B.54.i; E.173.B.54.i; E.174.B.54.i; E.175.B.54.i; E.176.B.54.i;
E.188.B.54.i;
E.189.B.54.i; E.190.B.54.i; E.196.B.54.i; E.202.B.54.i; E.205.B.54.i;
E.206.B.54.i;
E.207.B.54.i; E.208.B.54.i; E.209.B.54.i; E.210.B.54.i; E.211.B.54.i;
E.212.B.54.i;
E.213.B.54.i; E.700.B.54.i; E.701.B.54.i; E.702.B.54.i; E.703.B.54.i;
E.704.B.54.i;
E.705.B.54.i; E.706.B.54.i; E.707.B.54.i; E.708.B.54.i; E.709.B.54.i;
E.710.B.54.i;
E.711.B.54.i; E.712.B.54.i; E.713.B.54.i; E.714.B.54.i; E.715.B.54.i;
E.716.B.54.i;
E.717.B.54.i; E.718.B.54.i; E.719.B.54.i; E.720.B.54.i; E.721.B.54.i;
E.722.B.54.i;
E.723.B.54.i; E.724.B.54.i; E.3.B.54.o; E.4.B.54.o; E.7.B.54.o; E.9.B.54.o;
E.103.B.54.o;
E.106.B.54.o; E.107.B.54.o; E.108.B.54.o; E.111.B.54.o; E.114.B.54.o;
E.117.B.54.o;
E.118.B.54.o; E.119.B.54.o; E.120.B.54.o; E.121.B.54.o; E.137.B.54.o;
E.138.B.54.o;
E.139.B.54.o; E.140.B.54.o; E.141.B.54.o; E.142.B.54.o; E.145.B.54.o;
E.146.B.54.o;
E.147.B.54.o; E.148.B.54.o; E.149.B.54.o; E.150.B.54.o; E.151.B.54.o;
E.165.B.54.o;
E.166.B.54.o; E.167.B.54.o; E.168.B.54.o; E.169.B.54.o; E.170.B.54.o;
E.171.B.54.o;
E.172.B.54.o; E.173.B.54.o; E.174.B.54.o; E.175.B.54.o; E.176.B.54.o;
E.188.B.54.o;
E.189.B.54.o; E.190.B.54.o; E.196.B.54.o; E.202.B.54.o; E.205.B.54.o;
E.206.B.54.o;
E.207.B.54.o; E.208.B.54.o; E.209.B.54.o; E.210.B.54.o; E.211.B.54.o;
E.212.B.54.o;
E.213.B.54.o; E.700.B.54.o; E.701.B.54.o; E.702.B.54.o; E.703.B.54.o;
E.704.B.54.o;
E.705.B.54.o; E.706.B.54.o; E.707.B.54.o; E.708.B.54.o; E.709.B.54.o;
E.710.B.54.o;
E.711.B.54.o; E.712.B.54.o; E.713.B.54.o; E.714.B.54.o; E.715.B.54.o;
E.716.B.54.o;
E.717.B.54.o; E.718.B.54.o; E.719.B.54.o; E.720.B.54.o; E.721.B.54.o;
E.722.B.54.o;
- 98-
CA 02268756 1999-04-15
WO 98/17647 PCT/US97/18193
E.723.B.54.o; E.724.B.54.o; F.3.a.54.i; F.4.a.54.i; F.7.a.54.i; F.9.a.54.i;
F.103.a.54.i;
F.106.a.54.i; F.107.a.54.i; F.108.a.54.i; F.lll.a.54.i; F.114.a.54.i;
F.117.a.54.i;
F.118.a.54.i; F.119.a.54.i; F.120.a.54.i; F.121.a.54.i; F.137.a.54.i;
F.138.a.54.i;
F.139.a.54.i; F.140.a.54.i; F.141.a.54.i; F.142.a.54.i; F.145.a.54.i;
F.146.a.54.i;
F.147.a.54.i; F.148.a.54.i; F.149.a.54.i; F.150.a.54.i; F.151.a.54.i;
F.165.a.54.i;
F.166.a.54.i; F.167.a.54.i; F.168.a.54.i; F.169.a.54.i; F.170.a.54.i;
F.171.a.54.i;
F.172.a.54.i; F.173.a.54.i; F.174.a.54.i; F.175.a.54.i; F.176.a.54.i;
F.188.a.54.i;
F.189.a.54.i; F.190.a.54.i; F.196.a.54.i; F.202.a.54.i; F.205.a.54.i;
F.206.a.54.i;
F.207.a.54.i; F.208.a.54.i; F.209.a.54.i; F.210.a.54.i; F.211.a.54.i;
F.212.a.54.i;
F.213.a.54.i; F.700.a.54.i; F.701.a.54.i; F.702.a.54.i; F.703.a.54.i;
F.704.a.54.i;
F.705.a.54.i; F.706.a.54.i; F.707.a.54.i; F.708.a.54.i; F.709.a.54.i;
F.710.a.54.i;
F.711.a.54.i; F.712.a.54.i; F.713.a.54.i; F.714.a.54.i; F.715.a.54.i;
F.716.a.54.i;
F.717.a.54.i; F.718.a.54.i; F.719.a.54.i; F.720.a.54.i; F.721.a.54.i;
F.722.a.54.i;
F.723.a.54.i; F.724.a.54.i; F.3.a.54.o; F.4.a.54.o; F.7.a.54.o; F.9.a.54.o;
F.103.a.54.o;
F.106.a.54.o; F.107.a.54.o; F.108.a.54.o; F.111.a.54.o; F.114.a.54.o;
F.117.a.54.o;
F.118.a.54.o; F.119.a.54.o; F.120.a.54.o; F.121.a.54.o; F.137.a.54.o;
F.138.a.54.o;
F.139.a.54.o; F.140.a.54.o; F.141.a.54.o; F.142.a.54.o; F.145.a.54.o;
F.146.a.54.o;
F.147.a.54.o; F.148.a.54.o; F.149.a.54.o; F.150.a.54.o; F.151.a.54.o;
F.165.a.54.o;
F.166.a.54.o; F.167.a.54.o; F.168.a.54.o; F.169.a.54.o; F.170.a.54.o;
F.171.a.54.o;
F.172.a.54.o; F.173.a.54.o; F.174.a.54.o; F.175.a.54.o; F.176.a.54.o;
F.188.a.54.o;
F.189.a.54.o; F.190.a.54.o; F.196.a.54.o; F.202.a.54.o; F.205.a.54.o;
F.206.a.54.o;
F.207.a.54.o; F.208.a.54.o; F.209.a.54.o; F.210.a.54.o; F.211.a.54.o;
F.212.a.54.o;
F.213.a.54.o; F.700.a.54.o; F.701.a.54.o; F.702.a.54.o; F.703.a.54.o;
F.704.a.54.o;
F.705.a.54.o; F.706.a.54.o; F.707.a.54.o; F.708.a.54.o; F.709.a.54.o;
F.710.a.54.o;
F.711.a.54.o; F.712.a.54.o; F.713.a.54.o; F.714.a.54.o; F.715.a.54.o;
F.716.a.54.o;
F.717.a.54.o; F.718.a.54.o; F.719.a.54.o; F.720.a.54.o; F.721.a.54.o;
F.722.a.54.o;
F.723.a.54.o; F.724.a.54.o; F.172.b.54.i; F.173.b.54.i; F.174.b.54.i;
F.175.b.54.i;
F.176.b.54.i; F.188.b.54.i; F.189.b.54.i; F.190.b.54.i; F.196.b.54.i;
F.202.b.54.i;
F.205.b.54.i; F.206.b.54.i; F.207.b.54.i; F.208.b.54.i; F.209.b.54.i;
F.210.b.54.i;
F.211.b.54.i; F.212.b.54.i; F.213.b.54.i; F.700.b.54.i; F.701.b.54.i;
F.702.b.54.i;
F.703.b.54.i; F.704.b.54.i; F.705.b.54.i; F.706.b.54.i; F.707.b.54.i;
F.708.b.54.i;
F.709.b.54.i; F.710.b.54.i; F.711.b.54.i; F.712.b.54.i; F.713.b.54.i;
F.714.b.54.i;
F.715.b.54.i; F.716.b.54.i; F.717.b.54.i; F.718.b.54.i; F.719.b.54.i;
F.720.b.54.i;
F.721.b.54.i; F.722.b.54.i; F.723.b.54.i; F.724.b.54.i; F.3.b.54.o;
F.4.b.54.o; F.7.b.54.o;
F.9.b.54.o; F.103.b.54.o; F.106.b.54.o; F.107.b.54.o; F.108.b.54.o;
F.111.b.54.o;
F.114.b.54.o; F.117.b.54.o; F.118.b.54.o; F.119.b.54.o; F.120.b.54.o;
F.121.b.54.o;
F.137.b.54.o; F.138.b.54.o; F.139.b.54.o; F.140.b.54.o; F.141.b.54.o;
F.142.b.54.o;
F.145.b.54.o; F.146.b.54.o; F.147.b.54.o; F.148.b.54.o; F.149.b.54.o;
F.150.b.54.o;
F.151.b.54.o; F.165.b.54.o; F.166.b.54.o; F.167.b.54.o; F.168.b.54.o;
F.169.b.54.o;
F.170.b.54.o; F.171.b.54.o; F.172.b.54.o; F.173.b.54.o; F.174.b.54.o;
F.175.b.54.o;
F.176.b.54.o; F.188.b.54.o; F.189.b.54.o; F.190.b.54.o; F.196.b.54.o;
F.202.b.54.o;
F.205.b.54.o; F.206.b.54.o; F.207.b.54.o; F.208.b.54.o; F.209.b.54.o;
F.210.b.54.o;
F.211.b.54.o; F.212.b.54.o; F.213.b.54.o; F.700.b.54.o; F.701.b.54.o;
F.702.b.54.o;
F.703.b.54.o; F.704.b.54.o; F.705.b.54.o; F.706.b.54.o; F.707.b.54.o;
F.708.b.54.o;
F.709.b.54.o; F.710.b.54.o; F.711.b.54.o; F.712.b.54.o; F.713.b.54.o;
F.714.b.54.o;
F.715.b.54.o; F.716.b.54.o; F.717.b.54.o; F.718.b.54.o; F.719.b.54.o;
F.720.b.54.o;
F.721.b.54.o; F.722.b.54.o; F.723.b.54.o; F.724.b.54.o; F.172.x.54.i;
F.173.x.54.i;
F.174.x.54.i; F.175.x.54.i; F.176.x.54.i; F.188.x.54.i; F.189.x.54.i;
F.190.x.54.i;
F.196.x.54.i; F.202.x.54.i; F.205.x.54.i; F.206.x.54.i; F.207.x.54.i;
F.208.x.54.i;
F.209.x.54.i; F.210.x.54.i; F.211.x.54.i; F.212.x.54.i; F.213.x.54.i;
F.700.x.54.i;
F.701.x.54.i; F.702.x.54.i; F.703.x.54.i; F.704.x.54.i; F.705.x.54.i;
F.706.x.54.i;
F.707.x.54.i; F.708.x.54.i; F.709.x.54.i; F.710.x.54.i; F.711.x.54.i;
F.712.x.54.i;
F.713.x.54.i; F.714.x.54.i; F.715.x.54.i; F.716.x.54.i; F.717.x.54.i;
F.718.x.54.i;
F.719.x.54.i; F.720.x.54.i; F.721.x.54.i; F.722.x.54.i; F.723.x.54.i;
F.724.x.54.i; F.3.x.54.o;
F.4.x.54.o; F.7.x.54.o; F.9.x.54.o; F.103.x.54.o; F.106.x.54.o; F.107.x.54.o;
F.108.x.54.o;
F.111.x.54.o; F.114.x.54.o; F.117.x.54.o; F.118.x.54.o; F.119.x.54.o;
F.120.x.54.o;
- 99-
CA 02268756 1999-04-15
WO 98/17647 PCT/US97/18193
F.121.x.54.o; F.137.x.54.o; F.138.x.54.o; F.139.x.54.o; F.140.x.54.o;
F.141.x.54.o;
F.142.x.54.o; F.145.x.54.o; F.146.x.54.o; F.147.x.54.o; F.148.x.54.o;
F.149.x.54.o;
F.150.x.54.o; F.151.x.54.o; F.165.x.54.o; F.166.x.54.o; F.167.x.54.o;
F.168.x.54.o;
F.169.x.54.o; F.170.x.54.o; F.171.x.54.o; F.172.x.54.o; F.173.x.54.o;
F.174.x.54.o;
F.175.x.54.o; F.176.x.54.o; F.188.x.54.o; F.189.x.54.o; F.190.x.54.o;
F.196.x.54.o;
F.202.x.54.o; F.205.x.54.o; F.206.x.54.o; F.207.x.54.o; F.208.x.54.o;
F.209.x.54.o;
F.210.x.54.o; F.211.x.54.o; F.212.x.54.o; F.213.x.54.o; F.700.x.54.o;
F.701.x.54.o;
F.702.x.54.o; F.703.x.54.o; F.704.x.54.o; F.705.x.54.o; F.706.x.54.o;
F.707.x.54.o;
F.708.x.54.o; F.709.x.54.o; F.710.x.54.o; F.711.x.54.o; F.712.x.54.o;
F.713.x.54.o;
F.714.x.54.o; F.715.x.54.o; F.716.x.54.o; F.717.x.54.o; F.718.x.54.o;
F.719.x.54.o;
F.720.x.54.o; F.721.x.54.o; F.722.x.54.o; F.723.x.54.o; F.724.x.54.o;
F.172.y.54.i;
F.173.y.54.i; F.174.y.54.i; F.175.y.54.i; F.176.y.54.i; F.188.y.54.i;
F.189.y.54.i;
F.190.y.54.i; F.196.y.54.i; F.202.y.54.i; F.205.y.54.i; F.206.y.54.i;
F.207.y.54.i;
F.208.y.54.i; F.209.y.54.i; F.210.y.54.i; F.211.y.54.i; F.212.y.54.i;
F.213.y.54.i;
F.700.y.54.i; F.701.y.54.i; F.702.y.54.i; F.703.y.54.i; F.704.y.54.i;
F.705.y.54.i;
F.706.y.54.i; F.707.y.54.i; F.708.y.54.i; F.709.y.54.i; F.710.y.54.i;
F.711.y.54.i;
F.712.y.54.i; F.713.y.54.i; F.714.y.54.i; F.715.y.54.i; F.716.y.54.i;
F.717.y.54.i;
F.718.y.54.i; F.719.y.54.i; F.720.v.54.i; F.721.y.54.i; F.722.y.54.i;
F.723.y.54.i;
F.724.y.54.i; F.3.y.54.o; F.4.y.54.o; F.7.y.54.o; F.9.y.54.o; F.103.y.54.o;
F.106.y.54.o;
F.107.y.54.o; F.108.y.54.o; F.111.y.54.o; F.114.y.54.o; F.117.y.54.o;
F.118.y.54.o;
F.119.y.54.o; F.120.y.54.o; F.121.y.54.o; F.137.y.54.o; F.138.y.54.o;
F.139.y.54.o;
F.140.y.54.o; F.141.y.54.o; F.142.y.54.o; F.145.y.54.o; F.146.y.54.o;
F.147.y.54.o;
F.148.y.54.o; F.149.y.54.o; F.150.y.54.o; F.151.y.54.o; F.165.y.54.o;
F.166.y.54.o;
F.167.y.54.o; F.168.y.54.o; F.169.y.54.o; F.170.y.54.o; F.171.y.54.o;
F.172.y.54.o;
F.173.y.54.o; F.174.y.54.o; F.175.y.54.o; F.176.y.54.o; F.188.y.54.o;
F.189.y.54.o;
F.190.y.54.o; F.196.y.54.o; F.202.y.54.o; F.205.y.54.o; F.206.y.54.o;
F.207.y.54.o;
F.208.y.54.o; F.209.y.54.o; F.210.y.54.o; F.211.y.54.o; F.212.y.54.o;
F.213.y.54.o;
F.700.y.54.o; F.701.y.54.o; F.702.y.54.o; F.703.y.54.o; F.704.y.54.o;
F.705.y.54.o;
F.706.y.54.o; F.707.y.54.o; F.708.y.54.o; F.709.y.54.o; F.710.y.54.o;
F.711.y.54.o;
F.712.y.54.o; F.713.y.54.o; F.714.y.54.o; F.715.y.54.o; F.716.y.54.o;
F.717.y.54.o;
F.718.y.54.o; F.719.y.54.o; F.720.y.54.o; F.721.y.54.o; F.722.y.54.o;
F.723.y.54.o;
F.724.y.54.o; F.172.z.54.i; F.173.z.54.i; F.174.z.54.i; F.175.z.54.i;
F.176.z.54.i;
F.188.z.54.i; F.189.z.54.i; F.190.z.54.i; F.196.z.54.i; F.202.z.54.i;
F.205.z.54.i;
F.206.z.54.i; F.207.z.54.i; F.208.z.54.i; F.209.z.54.i; F.210.z.54.i;
F.211.z.54.i;
F.212.z.54.i; F.213.z.54.i; F.700.z.54.i; F.701.z.54.i; F.702.z.54.i;
F.703.z.54.i;
F.704.z.54.i; F.705.z.54.i; F.706.z.54.i; F.707.z.54.i; F.708.z.54.i;
F.709.z.54.i;
F.710.z.54.i; F.711.z.54.i; F.712.z.54.i; F.713.z.54.i; F.714.z.54.i;
F.715.z.54.i;
F.716.z.54.i; F.717.z.54.i; F.718.z.54.i; F.719.z.54.i; F.720.z.54.i;
F.721.z.54.i;
F.722.z.54.i; F.723.z.54.i; F.724.z.54.i; F.3.z.54.o; F.4.z.54.o; F.7.z.54.o;
F.9.z.54.o;
F.103.z.54.o; F.106.z.54.o; F.107.z.54.o; F.108.z.54.o; F.111.z.54.o;
F.114.z.54.o;
F.117.z.54.o; F.118.z.54.o; F.119.z.54.o; F.120.z.54.o; F.121.z.54.o;
F.137.z.54.o;
F.138.z.54.o; F.139.z.54.o; F.140.z.54.o; F.141.z.54.o; F.142.z.54.o;
F.145.z.54.o;
F.146.z.54.o; F.147.z.54.o; F.148.z.54.o; F.149.z.54.o; F.150.z.54.o;
F.151.z.54.o;
F.165.z.54.o; F.166.z.54.o; F.167.z.54.o; F.168.z.54.o; F.169.z.54.o;
F.170.z.54.o;
F.171.z.54.o; F.172.z.54.o; F.173.z.54.o; F.174.z.54.o; F.175.z.54.o;
F.176.z.54.o;
F.188.z.54.o; F.189.z.54.o; F.190.z.54.o; F.196.z.54.o; F.202.z.54.o;
F.205.z.54.o;
F.206.z.54.o; F.207.z.54.o; F.208.z.54.o; F.209.z.54.o; F.210.z.54.o;
F.211.z.54.o;
F.212.z.54.o; F.213.z.54.o; F.700.z.54.o; F.701.z.54.o; F.702.z.54.o;
F.703.z.54.o;
F.704.z.54.o; F.705.z.54.o; F.706.z.54.o; F.707.z.54.o; F.708.z.54.o;
F.709.z.54.o;
F.710.z.54.o; F.711.z.54.o; F.712.z.54.o; F.713.z.54.o; F.714.z.54.o;
F.715.z.54.o;
F.716.z.54.o; F.717.z.54.o; F.718.z.54.o; F.719.z.54.o; F.720.z.54.o;
F.721.z.54.o;
F.722.z.54.o; F.723.z.54.o; F.724.z.54.o; F.172.A.54.i; F.173.A.54.i;
F.174.A.54.i;
F.175.A.54.i; F.176.A.54.i; F.188.A.54.i; F.189.A.54.i; F.190.A.54.i;
F.196.A.54.i;
F.202.A.54.i; F.205.A.54.i; F.206.A.54.i; F.207.A.54.i; F.208.A.54.i;
F.209.A.54.i;
F.210.A.54.i; F.211.A.54.i; F.212.A.54.i; F.213.A.54.i; F.700.A.54.i;
F.701.A.54.i;
F.702.A.54.i; F.703.A.54.i; F.704.A.54.i; F.705.A.54.i; F.706.A.54.i;
F.707.A.54.i;
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WO 98/17647 PCT/US97/18193
F.708.A.54.i; F.709.A.54.i; F.710.A.54.i; F.711.A.54.i; F.712.A.54.i;
F.713.A.54.i;
F.714.A.54.i; F.715.A.54.i; F.716.A.54.i; F.717.A.54.i; F.718.A.54.i;
F.719.A.54.i;
F.720.A.54.i; F.721.A.54.i; F.722.A.54.i; F.723.A.54.i; F.724.A.54.i;
F.3.A.54.o;
F.4.A.54.o; F.7.A.54.o; F.9.A.54.o; F.103.A.54.o; F.106.A.54.o; F.107.A.54.o;
F.108.A.54.o; F.111.A.54.o; F.114.A.54.o; F.117.A.54.o; F.118.A.54.o;
F.119.A.54.o;
F.120.A.54.o; F.121.A.54.o; F.137.A.54.o; F.138.A.54.o; F.139.A.54.o;
F.140.A.54.o;
F.141.A.54.o; F.142.A.54.o; F.145.A.54.o; F.146.A.54.o; F.147.A.54.o;
F.148.A.54.o;
F.149.A.54.o; F.150.A.54.o; F.151.A.54.o; F.165.A.54.o; F.166.A.54.o;
F.167.A.54.o;
F.168.A.54.o; F.169.A.54.o; F.170.A.54.o; F.171.A.54.o; F.172.A.54.o;
F.173.A.54.o;
F.174.A.54.o; F.175.A.54.o; F.176.A.54.o; F.188.A.54.o; F.189.A.54.o;
F.190.A.54.o;
F.196.A.54.o; F.202.A.54.o; F.205.A.54.o; F.206.A.54.o; F.207.A.54.o;
F.208.A.54.o;
F.209.A.54.o; F.210.A.54.o; F.211.A.54.o; F.212.A.54.o; F.213.A.54.o;
F.700.A.54.o;
F.701.A.54.o; F.702.A.54.o; F.703.A.54.o; F.704.A.54.o; F.705.A.54.o;
F.706.A.54.o;
F.707.A.54.o; F.708.A.54.o; F.709.A.54.o; F.710.A.54.o; F.711.A.54.o;
F.712.A.54.o;
F.713.A.54.o; F.714.A.54.o; F.715.A.54.o; F.716.A.54.o; F.717.A.54.o;
F.718.A.54.o;
F.719.A.54.o; F.720.A.54.o; F.721.A.54.o; F.722.A.54.o; F.723.A.54.o;
F.724.A.54.o;
F.172.B.54.i; F.173.B.54.i; F.174.B.54.i; F.175.B.54.i; F.176.B.54.i;
F.188.B.54.i;
F.189.B.54.i; F.190.B.54.i; F.196.B.54.i; F.202.B.54.i; F.205.B.54.i;
F.206.B.54.i;
F.207.B.54.i; F.208.B.54.i; F.209.B.54.i; F.210.B.54.i; F.211.B.54.i;
F.212.B.54.i;
F.213.B.54.i; F.700.B.54.i; F.701.B.54.i; F.702.B.54.i; F.703.B.54.i;
F.704.B.54.i;
F.705.B.54.i; F.706.B.54.i; F.707.B.54.i; F.708.B.54.i; F.709.B.54.i;
F.710.B.54.i;
F.711.B.54.i; F.712.B.54.i; F.713.B.54.i; F.714.B.54.i; F.715.B.54.i;
F.716.B.54.i;
F.717.B.54.i; F.718.B.54.i; F.719.B.54.i; F.720.B.54.i; F.721.B.54.i;
F.722.B.54.i;
F.723.B.54.i; F.724.B.54.i; F.3.B.54.o; F.4.B.54.o; F.7.B.54.o; F.9.B.54.o;
F.103.B.54.o;
F.106.B.54.o; F.107.B.54.o; F.108.B.54.o; F.111.B.54.o; F.114.B.54.o;
F.117.B.54.o;
F.118.B.54.o; F.119.B.54.o; F.120.B.54.o; F.121.B.54.o; F.137.B.54.o;
F.138.B.54.o;
F.139.B.54.o; F.140.B.54.o; F.141.B.54.o; F.142.B.54.o; F.145.B.54.o;
F.146.B.54.o;
F.147.B.54.o; F.148.B.54.o; F.149.B.54.o; F.150.B.54.o; F.151.B.54.o;
F.165.B.54.o;
F.166.B.54.o; F.167.B.54.o; F.168.B.54.o; F.169.B.54.o; F.170.B.54.o;
F.171.B.54.o;
F.172.B.54.o; F.173.B.54.o; F.174.B.54.o; F.175.B.54.o; F.176.B.54.o;
F.188.B.54.o;
F.189.B.54.o; F.190.B.54.o; F.196.B.54.o; F.202.B.54.o; F.205.B.54.o;
F.206.B.54.o;
F.207.B.54.o; F.208.B.54.o; F.209.B.54.o; F.210.B.54.o; F.211.B.54.o;
F.212.B.54.o;
F.213.B.54.o; F.700.B.54.o; F.701.B.54.o; F.702.B.54.o; F.703.B.54.o;
F.704.B.54.o;
F.705.B.54.o; F.706.B.54.o; F.707.B.54.o; F.708.B.54.o; F.709.B.54.o;
F.710.B.54.o;
F.711.B.54.o; F.712.B.54.o; F.713.B.54.o; F.714.B.54.o; F.715.B.54.o;
F.716.B.54.o;
F.717.B.54.o; F.718.B.54.o; F.719.B.54.o; F.720.B.54.o; F.721.B.54.o;
F.722.B.54.o;
F.723.B.54.o; F.724.B.54.o.
Salts and Hydrates.
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++ and Mg++. 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 moiety, typically the W1 group carboxylic acid. Monovalent salts
are preferred if a water soluble salt is desired.
Metal salts typically are prepared by reacting the metal hydroxide with a
compound of this invention. Examples of metal salts which are prepared in
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WO 98/17647 PCT/US97/18193
this way are salts containing Li+, Na+, and K+. A less soluble metal salt can
be
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., HCI, HBr, H2SO4, H3P04, or organic sulfonic acids,
to
basic centers, typically amines of group G1, or to acidic groups such as E1.
Finally, it is to be understood that the compositions herein comprise
compounds of the invention in their un-ionized, as well as zwitterionic form,
and combinations with stoiochimetric 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
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.
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Methods of Inhibition of Neuraminidase.
Another aspect of the invention relates to methods of inhibiting the
activity of neuraminidase comprising the step of treating a sample suspected
of containing neuraminidase with a compound of the invention.
Compositions of the invention act as inhibitors of neuraminidase, 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
neuraminidase having a geometry unique to neuraminidase. Compositions
binding neuraminidase may bind with varying degrees of reversibility. Those
compounds binding substantially irreversibly are ideal candidates for use in
this method of the invention. In a typical embodiment the compositions bind
neuraminidase with a binding coefficient of less than 104M, more typically
less than 10-6M, still more typically 10-$M. Once labeled, the substantially
irreversibly binding compositions are useful as probes for the detection of
neuraminidase. Accordingly, the invention relates to methods of detecting
neuraminidase in a sample suspected of containing neuraminidase
comprising the steps of: treating a sample suspected of containing
neuraminidase 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 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 hydroxyl or amino.
Within the context of the invention samples suspected of containing
neuraminidase 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, 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 neuraminidase,
frequently a pathogenic organism such as a virus. Samples can be contained
in any medium 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
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WO 98/17647 PCT/US97/18193
composition to the sample. The addition step comprises any method of
administration as described above.
If desired, the activity of neuraminidase after application of the
composition can be observed by any method including direct and indirect
methods of detecting neuraminidase activity. Quantitative, qualitative, and
semiquantitative methods of determining neuraminidase activity are all
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 neuraminidase include bacteria (Vibrio
cholerae, Clostridium perfringens, Streptococcus pneumoniae, and
Arthrobacter sialophilus) and viruses (especially orthomyxoviruses or
paramyxoviruses such as influenza virus A and B, parainfluenza virus,
mumps virus, Newcastle disease virus, fowl plague virus, and sendai virus).
Inhibition of neuraminidase activity obtained from or found within any of
these organisms is within the objects of this invention. The virology of
influenza viruses is described in "Fundamental Virology" (Raven Press, New
York, 1986), Chapter 24. The compounds of this invention are useful in the
treatment or prophylaxis of such infections in animals, e.g. duck, rodents, or
swine, or in man.
However, in screening compounds capable of inhibiting influenza
viruses it should be kept in mind that the results of enzyme assays may not
correlate with cell culture assays, as shown Table 1 of Chandler et al., su
ra.
Thus, a plaque reduction assay should be the primary screening tool.
Screens for Neuraminidase Inhibitors.
Some of the compounds of the invention will be selective for particular
organisms such as bacterial verses viral neuraminidases or neuraminidase
from influenza verses parainfluenza. These compositions are identified by
routine screening.
Compositions of the invention are screened for inhibitory activity
against neuraminidase by any of the conventional techniques for evaluating
enzyme activity. Within the context of the invention, typically compositions
are first screened for inhibition of neuraminidase 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 5 X
10-6 M, typically less than about 1 X 10-7 M and preferably less than about 5
X
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WO 98/17647 PCT/US97/18193
10-8 M are preferred for in vivo use.
Useful in vitro screens have been described in detail and will not be
elaborated here. However, von Itzstein, M. et al. "Nature", 363(6428):418-423
(1993), in particular page 420, column 2, full paragraph 3, to page 421,
column
2, first partial paragraph, describes a suitable in vitro assay of Potier, M.
et al.
"Anaiyt. Biochem.", 94:287-296 (1979), as modified by Chong, A.K.J. et al.
"Biochem. Biophys. Acta", 1077:65-71 (1991); and Colman, P. M. et al.
International Publication No. WO 92/06691 (Int. App. No. PCT/AU90/00501,
publication date April 30, 1992) page 34, line 13, to page 35, line 16,
describes
another useful in vitro screen.
In vivo screens have also been described in detail, see von Itzstein, M.
et al. op. cit., in particular page 421, column 2, first full paragraph, to
page 423,
column 2, first partial paragraph, and Colman, P. M. et al. op. cit. page 36,
lines
1-38, describe suitable in vivo screens.
In screening assays used herein, compositions having IC,50 values
greater than 1 M (micromolar) are considered as being inactive against
influenza neuraminidase.
Pharmaceutical Formulations and Routes of Administration.
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
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). Excipients include ascorbic acid and other
antioxidants, chelating agents such as EDTA, carbohydrates such as dextrin,
hydroxyalkylcellulose, 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.
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 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
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invention is that they are orally bioavailable and can be dosed orally; it is
not
necessary to administer them by intrapulmonary or intranasal routes.
While it is possible for the active ingredients to be administered alone it
may be preferable to present them as pharmaceutical formulations. The
formulations, both for veterinary and for human use, of the invention
comprise at least one active ingredient, as above defined, together with one
or
more acceptable carriers therefor 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.
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). 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 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 invention suitable for oral administration are
prepared as discrete units such as capsules, cachets or tablets each
containing a
predetermined amount of the active ingredient; as a powder or granules; as
solution or a suspension in an aqueous liquid or a non-aqueous liquid; or as
an oil-in-water liquid emulsion or a water-in-oil liquid emulsion. The active
ingredient may also be presented as a bolus, electuary or paste.
A tablet is made by compression or molding, optionally with one or
more 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 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 therefrom. In one embodiment acid
hydrolysis of the medicament is obviated by use of an enteric coating.
For infections of the eye or other external tissues e.g. mouth and skin,
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the formulations are preferably applied as a topical ointment or cream
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 and 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 ointment base. Alternatively, the active ingredients may be
formulated in a cream with an oil-in-water cream base.
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 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 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-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, 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 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 Crodamol CAP may be used, the last three being preferred
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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.
Formulations suitable for topical administration to the eye also 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%, 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
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.
Formulations suitable for intrapulmonary or nasal administration have
a particle size for example in the range of 0.1 to 500 microns (including
particle
sizes in a range between 0.1 and 500 microns in increments microns such as
0.5, 1, 30 microns, 35 microns, 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 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 influenza A or B infections as described
below.
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 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,
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for 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 are those containing a daily dose or unit daily sub-dose, as
herein above recited, or an appropriate fraction thereof, of the active
ingredient.
It should be understood that in addition to the ingredients particularly
mentioned above 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.
The invention further provides veterinary compositions comprising at
least one active ingredient as above defined together with a veterinary
carrier
therefor.
Veterinary carriers are materials useful for the purpose of
administering the composition and may be solid, liquid or gaseous materials
which are 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 are used to provide controlled release
pharmaceutical formulations containing as active ingredient one or more
compounds of the invention ("controlled release formulations") in which the
release of the active ingredient are controlled and regulated to allow less
frequency dosing or to improve the pharmacokinetic or toxicity profile of a
given active ingredient.
Effective dose of active ingredient depends at least on the nature of the
condition being treated, toxicity, whether the compound is being used
prophylactically (lower doses) or against an active influenza infection, the
method of delivery, and the pharmaceutical formulation, and will be
determined by the clinician using conventional dose escalation studies. It can
be expected to be from about 0.0001 to about 100 mg/kg body weight per day.
Typically, from about 0.01 to about 10 mg/kg body weight per day. More
typically, from about.01 to about 5 mg/kg body weight per day. More typically,
from about .05 to about 0.5 mg/kg body weight per day. For example, for
inhalation the daily candidate dose for an adult human of approximately 70 kg
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body weight will range from 1 mg to 1000 mg, preferably between 5 mg and 500
mg, and may take the form of single or multiple doses.
Active ingredients of the invention are also used in combination with
other active ingredients. Such combinations are selected based on the
condition to be treated, cross-reactivities of ingredients and pharmaco-
properties of the combination. For example, when treating viral infections of
the respiratory system, in particular influenza infection, the compositions of
the invention are combined with antivirals (such as amantidine, rimantadine
and ribavirin), mucolytics, expectorants, bronchialdilators, antibiotics,
antipyretics, or analgesics. Ordinarily, antibiotics, antipyretics, and
analgesics
are administered together with the compounds of this invention.
Enteric Protection
Another embodiment of the present invention is directed toward
enteric protected forms of the compounds of the invention. As used herein
the term "enteric protection" means protecting a compound of the invention
in order to avoid exposing a portion of the gastrointestinal tract, typically
the
upper gastrointestinal tract, in particular the stomach and esophagus, to the
compound of this invention. In this way gastric mucosal tissue is protected
against rates of exposure to a compound of the invention which produce
adverse effects such as nausea; and, alternatively, a compound of the
invention is protected from conditions present in one or more portions of the
gastrointestinal tract, typically the upper gastrointestinal tract.
By way of example and not limitation, such enterically protected forms
include enteric coated vehicles, such as enteric coated tablets, enteric
coated
granules, enteric coated beads, enteric coated particles, enteric coated
microparticles, and enteric coated capsules. In preferred embodiments, a
compound of the invention is placed in a suitable vehicle such as a tablet,
granule or capsule, and the vehicle is coated with a pharmaceutically
acceptable enteric coating. In alternative preferred embodiments, a compound
of the invention is prepared as enterically protected granules, particles,
microparticles, spheres, microspheres, or colloids, and the enteric protected
granules, particles, microparticles, spheres, microspheres, or colloids, are
prepared as pharmaceutically acceptable dosage forms such as tablets,
granules,
capsules, or suspensions.
One aspect of the invention is directed to enteric-coated dosage forms of
the compounds of the invention to effect delivery to the intestine of a human
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or other mammal, preferably to the small intestine, of a pharmaceutical
composition comprised of a therapeutically effective amount of about 0.1-1000
mg of an active ingredient and optional pharmaceutically acceptable
excipients.
The term "vehicle" as used herein includes pharmaceutically acceptable
dose vehicles. Many vehicles are well known in the art cited herein such as
tablet, coated tablet, capsule, hard capsule, soft gelatin capsule, particle,
microparticle, sphere, microsphere, colloid, microencapsulationed, sustained
release, semisolid, suppository or granule vehicles.
The term "pharmaceutically-acceptable excipients" as used herein
includes any physiologically inert, pharmacologically inactive material known
to one skilled in the art, which is compatible with the physical and chemical
characteristics of the particular compound of the invention selected for use.
These excipients are described elsewhere herein. The excipients may, but need
not, provide enteric protection.
The term "unit dose" is used herein in the conventional sense to mean
a single application or administration of the compound of this invention to
the subject being treated in an amount as stated below. It should be
understood that a therapeutic or prophylactic dosage can be given in one unit
dose, or alternatively, in multiples of two or more of such dose units with
the
total adding up to the desired amount of compound for a given time period.
In general, the oral unit dosage form compositions of this invention,
preferably employ from about 1 to about 1000 milligrams (mg), typically, about
10 to 500 mg, more typically from about 50 to about 300 mg, more typically
yet,
75 mg of the compound for each unit dose. The actual amount will vary
depending upon the active compound selected.
In typical embodiments, an enteric protectant is applied to the vehicle
containing the compound, or to the compound without vehicle, the
protectant prevents nausea inducing exposure, contact or rates of exposure of
the mouth, esophagus or stomach with the compound, but which releases the
compound for absorption when the dosage form passes into the proximal
portion of the lower gastrointestinal tract, or in some embodiments,
substantially only in the colon.
The relative proportions of the protectant and compound of the
invention are varied to achieve optimum absorption, depending on the
compound selected. The minimum or maximum amount of enteric
protectant by weight percent is not critical. Typically, enteric protected
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embodiments contain less than about 50% enteric coating by weight. More
typically about 1% to about 25%, still more typically, about 1% to about 15%,
more typically yet, about 1% to about 10% (all by weight).
Related Art:
A number of monographs describe enteric protection and related
technology. Such monographs include: "Theory and Practice of Industrial
Pharmacy," 3rd ed. Lea & Febiger, Philadelphia, 1986 (ISBN 0-8121-0977-5);
Lehmann, K.; "Practical Course in Laquer Coating,", Eudragit, 1989; Lieberman;
Lachman, L.; Schwartz, "Pharmaceutical Dosage Forms: Tablets", 1990, Dekker
(ISBN: 0-8247-8289-5); Lee, Ping I. Editor Good, William R. Editor,
"Controlled-
Release Technology: Pharmaceutical Applications", ACS Symposium Ser.Vol.
348 (ISBN: 0-608-03871-7); Wilson, Billie E., Shannon, Margaret T., "Dosage
Calculation: A Simplified Approach", 1996, Appleton & Lange (ISBN: 0-8385-
9297-X); Lieberman, Herbert A. Editor Rieger, Martin M., "Pharmaceutical
Dosage Forms - Disperse Systems", 1996, Dekker (ISBN: 0-8247-9387-0); "Basic
Tests for Pharmaceutical Dosage Forms", 1995, World Health (ISBN: 92-4-
154418-X); Karsa, D. R., Editor; Stephenson, R. A., Editor, "Excipients &
Delivery Systems for Pharmaceutical Formulations: Proceedings of the
"Formulate '94" British Association for Chemical Specialties Symposium",
1995, CRC Pr (ISBN: 0-85404-715-8); Ansel, Howard C.; Popovich, Nicholas G.;
Allen, Lloyd V., "Pharmaceutical Dosage Forms & Drug Delivery Systems, 6th
ed.", 1994, Williams & Wilkins (ISBN: 0-683-01930-9); "The Sourcebook for
Innovative Drug Delivery: Manufacturers of Devices & Pharmaceuticals,
Suppliers of Products & Services, Sources of Information", 1987, Canon
Comns (ISBN: 0-9618649-0-7); Chiellini, E., Editor; Giusti, G., Editor;
Migliaresi,
C., Editor; Nicolais, L., Editor, "Polymers in Medicine II: Biomedical &
Pharmaceutical Applications", 1986, Plenum (ISBN: 0-306-42390-1);
"Pharmaceutical Aerosol: A Drug Delivery System in Transition", 1994,
Technomic (ISBN: 0-87762-971-4); Avid; Lieberman, L.; Lachman,
"Pharmaceutical Dosage Forms: Parenteral Medication, 2nd Expanded;
Revised ed.", 1992, Dekker (ISBN: 0-8247-9020-0); Laffer, U., Editor;
Bachmann,
I., Editor; Metzger, U., Editor, "Implantable Drug Delivery Systems", 1991, S
Karger (ISBN: 3-8055-5434-6); Borchardt, Ronald T., Editor; Repta, Arnold J.,
Editor; Stella, Valentino J., Editor, "Directed Drug Delivery: A
Multidisciplinary Approach", 1985, Humana (ISBN: 0-89603-089-X); Anderson,
James M., Editor, "Advances in Drug Delivery Systems 5: Proceedings of the
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Fifth International Symposium on Recent Advances in Drug Delivery
Systems, Salt Lake City, UT, U. S. A., February 25-28, 1991", Elsevier (ISBN:
0-
444-88664-8); Turco, Salvatore J.; King, Robert E., "Sterile Dosage Forms:
Their
Preparation & Clinical Application", 1987, Williams & Wilkins (ISBN: 0-8121-
1067-6); Tomlinson, E., Editor; Davis, S. S., Editor, "Site-Specific Drug
Delivery:
Cell Biology, Medical & Pharmaceutical Aspects", 1986, Wiley (ISBN: 0-471-
91236-0); Hess, H., Editor, "Pharmaceutical Dosage Forms & Their Use", 1986,
Hogrefe & Huber Pubs (ISBN: 3-456-81422-4); Avis; Lieberman; Lachman,
"Pharmaceutical Dosage Forms, Vol. 2", 1986, Dekker (ISBN: 0-8247-7085-4);
Carstensen, Jens T., "Pharmaceutics of Solids & Solid Dosage Forms", 1977,
Wiley (ISBN: 0-471-13726-X); Robinson, Joseph R., Editor, "Ophthalmic Drug
Delivery Systems", 1980, Am Pharm Assn (ISBN: 0-917330-32-3); Ansel,
Howard C., "Introduction to Pharmaceutical Dosage Forms, 4th ed.", 1985,
Williams & Wilkins (ISBN: 0-8121-0956-2); "High Tech Drug Delivery
Systems", 1984, Intl Res Dev (ISBN: 0-88694-622-0); Swarbrick, James, "Current
Concepts in Pharmaceutical Sciences: Dosage Form Design & Bioavailability",
1985, Lea & Febiger (ISBN: 0-318-79917-0); Sprowls, Joseph B., Editor,
"Prescription Pharmacy: Dosage Formulation & Pharmaceutical Adjuncts, 2nd
ed.", 1970, Lippincott (ISBN: 0-397-52050-6); and Polderman, J., Editor,
"Formulation & Preparation of Dosage Forms: Proceedings of the 37th
International Congress of Pharmaceutical Sciences of F.I.P., The Hague,
Netherlands, September, 1977", Elsevier (ISBN: 0-444-80033-6).
Specific Embodiments:
In another embodiment, the inventive composition is in the form of an
enteric coated tablet dosage form. In this embodiment, the formulation is
formed into a hard tablet by conventional means and the tablet is coated with
the enteric coating in accordance with conventional techniques.
In a preferred embodiment, the inventive compound is in the form of
an enteric coated powder dosage form. In this embodiment, the formulation
is filled into a hard or soft-shell capsule or their equivalent and the
capsule is
coated with the enteric coating in accordance with conventional techniques.
In one embodiment the inventive composition is in the form of a
liquid suspension of enteric coated particles of a compound of the invention.
In this embodiment, a suspension of the inhibitor in a liquid is filled into a
hard or soft-shell capsule or their equivalent and the capsule is coated with
the
enteric coating in accordance with conventional techniques.
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As alternatives to the foregoing embodiments the capsule or other
dosage container is itself constructed of an enteric protection reagent or
component, or otherwise is integral to the container.
In another embodiment enteric protectants are used to administer a
compound of the invention to the colon. The delivery system is a tablet
comprised of three layers: 1) a core containing the active compound of the
invention; 2) a non-swelling, erodible polymer layer surrounding the core
(with the combination of core and erodible polymer layer being referred to as
the "dual matrix tablet"); and 3) an enteric coating applied to the dual
matrix
tablet. The composition and function of the components of such a colon
targeted -aelivery system are further described in U.S. Patent 5,482,718.
Another embodiment of the invention is directed toward enteric
protected emulsion, suspension, tablet, coated tablet, hard capsule, soft
gelatin
capsule, microencapsulation, sustained release, liquid, semisolid,
suppositorie,
and aerosol dosage forms of the compounds of_the invention. "Theory and
Practice of Industrial Pharmacy," 3rd ed. Lea & Febiger, Philadelphia, 1986
(ISBN 0-8121-0977-5), describes each of these standard dosage forms in detail
at
the following locations: emulsion and suspension dosage forms (pp. 100-122),
tablets (pp. 293-345), coated tablet (pp. 346-373), hard capsules (pp. 374-
397), soft
gelatin capsules (pp. 398-411), microencapsulation (pp. 412-430), sustained
release dosage forms (pp. 430-456), liquids (pp. 457-478), pharmaceutical
suspensions (pp. 479-501), emulsions (pp. 502-533), semisolids (pp. 534-563),
suppositories (pp. 564-587), and pharmaceutical aerosols (pp. 589-618).
Alternative embodiments include enteric protected sustained release,
controlled release, particulate, microencapsulated, multip articulate,
microparticulate, colloidal, nasal, inhalation, oral mucosal, colonic, dermal,
transdermal, ocular, topical, and veterinary dosage forms of the compounds of
the invention.
Materials:
Conventional enteric protectant polymers or mixtures of polymers for
use herein include insoluble at a pH below about 5.5, i.e., that which is
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generally found in the stomach, but are soluble at pH about 5.5 or above,
i.e.,
that present in the small intestine and the large intestine. The effectiveness
of
particular enteric protectant materials can be measured using known USP
procedures.
Exemplary enteric protectant polymers employable in this embodiment
include cellulose acetate phthalate, methyl acrylate-methacrylic acid
copolymers, cellulose acetate succinate, hydroxypropylmethylcellulose
phthalate, polyvinyl acetate phthalate, and methyl methacrylate-methacrylic
acid copolymers. Another example is an anionic carboxylic copolymers based
on methacrylic acid and methacrylate, commercially available as Eudragit(r)*
Typical examples include cellulose acetate phthalate ("CAP"), cellulose
acetate
trimellitate, hydroxypropyl methylcellulose phthalate ("HPMCP"),
hydroxypropyl methylcellulose phthalate succinate, polyvinyl acetate
phthalate ("PVAP"), methacrylic acid, and methacrylic acid esters. More
typically the protectant is selected from, PVAP and/or HPMCP, particularly
PVAP. PVAP is known under the trademark Sureteric(r), manufactured by
Colorcon, Inc.
The enteric protectant materials may be applied to the vehicle with or
without conventional plasticizers, such as acetylated mono glycerides,
propylene glycol, glycerol, glyceryl triacetate, polyethylene glycol, triethyl
citrate, tributyl citrate, diethyl phthalate, or dibutyl phthalate using
methods
known to those skilled in the art.
Exemplary Embodiments of Enteric Protection:
Embodiment 1: Enteric Protected A 141 x 4 i Capsules
In this exemplary embodiment, compound A.141.x.4.i, phosphate salt
form, 100 mg/capsule, is mixed with Croscarmellose Sodium (2.6 mg/capsule)
in a size 4 white opaque hard gelatin capsule shells (capsule composition:
gelatin NF, titanium dioxide USP) and the capsule is enterically coated.
The following enteric coating formulations are applied to the capsule by
procedures known to those in the art.
Ingredients % w/w
Preparation A:
Hydroxypropyl methylcellulose phthalate ("HPMCP")
* trademarks 115
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5.0
Triacetin 0.5
Alcohol USP 7.9
Water 15.5
Preparation B:
HPMCP 10.0
Titanium dioxide 0.2
Dimethyl polysiloxane 0.05
Triethyl citrate 1.0
Alcohol USP 72.75
Water 16.00
Preparation C:
Cellulose acetate phthalate ("CAP")
8.5
Diethyl phthalate 1.5
Titanium dioxide 0.2
Acetone 44.9
Denatured alcohol 44.9
Preparation D:
Polyvinyl "acetate phthalate ("PVAP")
5.0
Acetylated glycerides 0.8
Methylene chloride 47.1
Denatured alcohol 47.1
Prenaration E:
_
Methacrylic acid or methacrylic
8.0
acid ester (Eudragit (r7S or L, manufactured by
Rohm Pharma, GMBH, Wetterstadt,
West Germany)
Acetone 46.0
Arihydrous alcohol 46.0
Plasticizer q.s.
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Typically the enteric polymer (with or without plasticizer) is dissolved
in the solvents described under each formulation to form a
suspension/solution. Optionally, an opacifer such as titanium dioxide is
added. The vehicle is sprayed with the coating suspension/solution in a
suitable vessel under conditions such that an enterically-protected coating is
laid down on the vehicle without dissolving or disrupting the vehicle.
Approximately 1-50%, typically 1-15%, more typically, 5-10% by weight of the
finished coated vehicle of the enteric polymer coating will be useful for
adequate enteric protection.
Embodiment 2: Enteric Protected Tablet
In another exemplary embodiment a core tablet is encased within an
enteric coating. Optionally, a subcoating is used.
Core Tablets:
Core tablets of the present invention may be formed by combining (a)
the active ingredient with pharmaceutically-acceptable excipients in a mixture
including for example: a diluent, a binder, a disintegrant, and optionally one
or more ingredients selected from a group consisting of: compression aids,
flavors, flavor enhancers, sweeteners, dyes, pigments, buffer systems, and
preservatives; (b) lubricating the mixture with a lubricant; and (c)
compressing
the resultant lubricated mixture into a desired tablet form using various
tableting techniques available to those skilled in the art. The term "tablet"
as
used herein is intended to encompass compressed or formed pharmaceutizal
dosage formulations of all shapes and sizes.
Typical diluents employable in this embodiment include lactose or
microcrystalline cellulose.
Typical binders employable in this embodiment include, but are not
limited to, povidone. Povidone is available under the trade name "Avicel"
from ISP Corporation.
The disintegrant may be one of several modified starrhes, or modified
cellulose polymers. Typically, croscarmellose sodium is used.
Croscarmellose*sodium NF Type A is commercially available under the trade
name "Ac-di-sol".
Typical lubricants include magnesium stearate, stearic acid,
hydrogenated vegetable oil or talc.
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Flavoring agents include those described in Remington's
Pharmaceutical Sciences, 18th Edition, Mack Publishing Company, 1990, pp.
1288-1300.
Typical sweeteners include saccharin, Aspartame, or edible mono- or
disaccharides such as glucose or sucrose.
Dyes and pigments include those described in the Handbook of
Pharmaceutical Excipients, pp. 81-90, 1986 by the American Pharmaceutical
Association & the Pharmaceutical Society of Great Britain.
Typical preservatives include methyl paraben, propyl paraben,
cetylpyridinium chloride, and the salts thereof, sorbic acid and the salts
thereof, thimerosal, or benzalkonium chloride.
Enteric Coating:
Eudragit'"L-30-D(r), a methacrylic acid copolymer, manufactured by
Rohm Pharma GmbH, Weiterstadt, West Germany, is a suitable enteric
polymer. Eudragit L-30-D(r) has a ratio of free carboxyl groups to ester
groups
of approximately 1:1 and is freely soluble at pH 5.5 and above. In general,
the
greater the percentage of Eudragit L-30-D(r) contained in the enteric coating,
the more proximal the release of active in the lower gastrointestinal tract.
The
location in the lower gastrointestinal tract at which the coating releases the
compound can be manipulated by one skilled in the art through control of the
composition and thickness of the applied enteric coating.
Typically a plasticizer, such as those set forth above, is included. Other
additives such as talc or silica may be used as detackifiers to improve the
coating process.
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Subcoating:
Optionally a stability enhancing subcoat on the core tablet is used to
minimize interaction between the compound of this invention and the
enteric coating. This also permits utilization of a single 10-300 micron thick
enteric film without affecting product stability. This subcoat inhibits
migration of active ingredient from the core tablet into the enteric coating,
thus improving shelf life and product stability, but the subcoat rapidly
dissolves in intestinal fluid once the exterior enteric coating has been
breached.
Typical subcoating polymers employable in this embodiment include
hydroxypropyl methylcellulose, hydroxypropyl cellulose, hydroxypropyl
ethylcellulose, or polyvinylpyrrolidone.
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 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. Such products typically are identified by
preparing a radiolabelled (e.g. C14 or H3) compound of the invention,
administering it parenterally in a detectable 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
hours) and isolating its conversion products from the urine, blood or other
biological samples. These products are easily isolated since they are labeled
30 (others are isolated by the use of antibodies 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 assays for
therapeutic dosing of the compounds of the invention even if they possess no
neuraminidase inhibitory activity of their own.
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Additional Uses for the Compounds of This Invention.
The compounds of this invention, or the biologically active substances
produced from these compounds by hydrolysis or metabolism in vivo, are
used as immunogens or for conjugation to proteins, whereby they serve as
components of immunogenic compositions to prepare antibodies capable of
binding specifically to the protein, to the compounds or to their metabolic
products which retain immunologically recognized epitopes (sites of antibody
binding). The immunogenic compositions therefore are useful as
intermediates in the preparation of antibodies for use in diagnostic, quality
control, or the like, methods or in assays for the compounds or their novel
metabolic products. The compounds are useful for raising antibodies against
otherwise non-immunogenic polypeptides, in that the compounds serve as
haptenic sites stimulating an immune response that cross-reacts with the
unmodified conjugated protein.
The hydrolysis products of interest include products of the hydrolysis of
the protected acidic and basic groups discussed above. As noted above, the
acidic or basic amides comprising immunogenic polypeptides such as albumin
or keyhole limpet hemocyanin generally are useful as immunogens. The
metabolic products described above may retain a substantial degree of
immunological cross reactivity with the compounds of the invention. Thus,
the antibodies of this invention will be capable of binding to the unprotected
compounds of the invention without binding to the protected compounds;
alternatively the metabolic products, will be capable of binding to the
protected
compounds and/or the metabolitic products without binding to the protected
compounds of the invention, or will be capable of binding specifically to any
one or all three. The antibodies desirably will not substantially cross-react
with naturally-occurring materials. Substantial cross-reactivity is reactivity
under specific assay conditions for specific analytes sufficient to interfere
with
the: assay results.
The immunogens of this invention contain the compound of this
invention presenting the desired epitope in association with an immunogenic
substance. Within the context of the invention such association means
covalent bonding to form an immunogenic conjugate (when applicable) or a
mixture of non-covalently bonded materials, or a combination of the above.
Immunogenic substances include adjuvants such as Freund's adjuvant,
immunogenic proteins such as viral, bacterial, yeast, plant and animal
polypeptides, in particular keyhole limpet hemocyanin, serum albumin,
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bovine thyroglobulin or soybean trypsin inhibitor, and immunogenic
polysaccharides. Typically, the compound having the structure of the desired
epitope is covalently conjugated to an immunogenic polypeptide or
polysaccharide by the use of a polyfunctional (ordinarily bifunctional) cross-
linking agent. Methods for the manufacture of hapten immunogens are
conventional e}~ r se, and any of the methods used heretofore for conjugating
haptens to immunogenic polypeptides or the like are suitably employed here
as well, taking into account the functional groups on the precursors or
hydrolytic products which are available for cross-linking and the likelihood
of
producing antibodies specific to the epitope in question as opposed to the
immunogenic substance.
Typically the polypeptide is conjugated to a site on the compound of the
invention distant from the epitope to be recognized.
The conjugates are prepared in conventional fashion. For example, the
cross-linking agents N-hydroxysuccinimide, succinic anhydride or
alkN=C=Nalk are useful in preparing the conjugates of this invention. The
conjugates comprise a compound of the invention attached by a bond or a
linking group of 1-100, typically, 1-25, more typically 1-10 carbon atoms to
the
immunogenic substance. The conjugates are separated from starting materials
and by products using chromatography or the like, and then are sterile
filtered
and vialed for storage.
The compounds of this invention are cross-linked for example through
any one or more of the following groups: a hydroxyl group of W6; a carboxyl
group of El; a carbon atom of W6, E1, G1, or Tl, in substitution of H; and an
amine group of G1. Included within such compounds are amides of
polypeptides where the polypeptide serves as an above-described R6c or R6b
groups.
Animals are typically immunized against the immunogenic conjugates
or derivatives and antisera or monoclonal antibodies prepared in
conventional fashion.
The compounds of the invention are useful for maintaining the
structural integrity of glycoproteins in recombinant cell culture, i.e., they
are
added to fermentations in which glycoproteins are being produced for
recovery so as to inhibit neuraminidase-catalyzed cleavage of the desired
glycoproteins. This is of particular value in the recombinant synthesis of
proteins in heterologous host cells that may disadvantageously degrade the
carbohydrate portion of the protein being synthesized.
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The compounds of the invention are polyfunctional. As such they
represent a unique class of monomers for the synthesis of polymers. By way of
example and not limitation, the polymers prepared from the compounds of
this invention include polyamides and polyesters.
The present compounds are used as monomers to provide access to
polymers having unique pendent functionalities. The compounds of this
invention are useful in homopolymers, or as comonomers with monomers
which do not fall within the scope of the invention. Homopolymers of the
compounds of this invention will have utility as cation exchange agents
(polyesters or polyamides) in the preparation of molecular sieves
(polyamides), textiles, fibers, films, formed articles and the like where the
acid
functionality El is esterified to a hydroxyl group in W6, for example, whereby
the pendant basic group G1 is capable of binding acidic functionalities such
as
are found in polypeptides whose purification is desired. Polyamides are
prepared by cross-linking El and G1, with W6 and the adjacent portion of the
ring remaining free to function as a hydrophilic or hydrophobic affinity
group,
depending up the selection of the W6 group. The preparation of these
polymers from the compounds of the invention is conventional ep r se.
The compounds of the invention are also useful as a unique class of
polyfunctional surfactants. Particularly when W6 does not contain a
hydrophilic substituent and is, for example, alkyl or alkoxy, the compounds
have the properties of bi-functional surfactants. As such they have useful
surfactant, surface coating, emulsion modifying, rheology modifying and
surface wetting properties.
As polyfunctional compounds with defined geometry and carrying
simultaneously polar and non-polar moieties, the compounds of the
invention are useful as a unique class of phase transfer agents. By way of
example and not limitation, the compounds of the invention are useful in
phase transfer catalysis and liquid/liquid ion extraction (LIX).
The compounds of the invention optionally contain asymmetric carbon
atoms in groups W6, E1, Gi, and T1. As such, they are a unique class of chiral
auxiliaries for use in the synthesis or resolution of other optically active
materials. For example, a racemic mixture of carboxylic acids can be resolved
into its component enantiomers by: 1) forming a mixture of diastereomeric
esters or amides with a compound of the invention wherein W6 is an
asymmetric hydroxyalkane or amino alkane group; 2) separating the
diastereomers; and 3) hydrolyzing the ester structure. Racemic alcohols are
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separated by ester formation with an acid group of El. Further, such a method
can be used to resolve the compounds of the invention themselves if optically
active acids or alcohols are used instead of racemic starting materials.
The compounds of this invention are useful as linkers or spacers in
preparing affinity absorption matrices, immobilized enzymes for process
control, or immunoassay reagents. The compounds herein contain a
multiplicity of functional groups that are suitable as sites for cross-linking
desired substances. For example, it is conventional to link affinity reagents
such as hormones, peptides, antibodies, drugs, and the like to insoluble
substrates. These insolublized reagents are employed in known fashion to
absorb binding partners for the affinity reagents from manufactured
preparations, diagnostic samples and other impure mixtures. Similarly,
immobilized enzymes are used to perform catalytic conversions with facile
recovery of enzyme. Bifunctional compounds are commonly used to link
analytes to detectable groups in preparing diagnostic reagents.
Many functional groups in the compounds of this invention are
suitable for use in cross-linking. For example, the carboxylic or phosphonic
acid of group El is used to form esters with alcohols or amides with amines of
the reagent to be cross-linked. The Gl sites substituted with OH, NHR1, SH,
azido (which is reduced to amino if desired before cross-linking), CN, NO2,
amino, guanidino, halo and the like are suitable sites. Suitable protection of
reactive groups will be used where necessary while assembling the cross-
linked reagent to prevent polymerization of the bifunctional compound of
this invention. In general, the compounds here are used by linking them
through carboxylic or phosphonic acid to the hydroxyl or amino groups of the
first linked partner, then covalently bonded to the other binding partner
through a Tl or G1 group. For example a first binding partner such as a
steroid
hormone is esterified to the carboxylic acid of a compound of this invention
and then this conjugate is cross-linked through a G1 hydroxyl to cyanogen
bromide activated Sepaharose, whereby immobilized steroid is obtained.
Other chemistries for conjugation are well known. See for example Maggio,
"Enzyme-Immunoassay" (CRC, 1988, pp 71-135) and references cited therein.
As noted above, the therapeutically useful compounds of this
invention in which the W1, or G1 carboxyl, hydroxyl or amino groups are
protected are useful as oral or sustained release forms. In these uses the
protecting group is removed in vivo, e.g., hydrolyzed or oxidized, so as to
yield
the free carboxyl, amino or hydroxyl. Suitable esters or amides for this
utility
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are selected based on the substrate specificity of esterases and/or
carboxypeptidases expected to be found within cells where precursor hydrolysis
is desired. To the extent that the specificity of these enzymes is unknown,
one
will screen a plurality of the compounds of this invention until the desired
substrate specificity is found. This will be apparent from the appearance of
free
compound or of antiviral activity. One generally selects amides or esters of
the invention compound that are (i) not hydrolyzed or hydrolyzed
comparatively slowly in the upper gut, (ii) gut and cell permeable and (iii)
hydrolyzed in the cell cytoplasm and/or systemic circulation. Screening assays
preferably use cells from particular tissues that are susceptible to influenza
infection, e.g. the mucous membranes of the bronchopulmonary tract. Assays
known in the art are suitable for determining in vivo bioavailability
including
intestinal lumen stability, cell permeation, liver homogenate stability and
plasma stability assays. However, even if the ester, amide or other protected
derivatives are not converted in vivo to the free carboxyl, amino or hydroxyl
groups, they remain useful as chemical intermediates.
Exemplary Methods of Making the Compounds of the Invention.
The invention also relates to 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. However, many of the known techniques are elaborated in
"Compendium of Organic Synthetic Methods" (John Wiley & Sons, New
York), Vol. 1, Ian T. Harrison and Shuyen Harrison, .1971; Vol. 2, Ian 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", (John Wiley & Sons, New York, 1985),
"Comprehensive Organic Synthesis. Selectivity, Strategy & Efficiency in
Modern Organic Chemistry. In 9 Volumes", Barry M. Trost, Editor-in-Chief
(Pergamon Press, New York, 1993 printing).
A number of exemplary methods for the preparation of the
compositions 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.
Generally, the reaction conditions such as temperature, reaction time,
solvents, workup procedures, and the like, will be those common in the art for
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the particular reaction to be performed. The cited reference material,
together
with material cited therein, contains detailed descriptions of such
conditions.
Typically the temperatures will be -100 C to 200 C, solvents will be aprotic
or
protic, and reaction times will be 10 seconds to 10 days. Workup typically
consists of quenching any unreacted reagents followed by partition between a
water/organic layer system (extraction) and separating the layer containing
the
product.
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 C to -100 C, solvents
are
typically aprotic for reductions and may be either protic or aprotic for
oxidations. Reaction times are adjusted to achieve desired conversions.
Condensation reactions are typically carried out at temperatures near
room temperature, although for non-equilibrating, kinetically controlled
condensations reduced temperatures (0 C to -100 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
by-products and use of anhydrous reaction conditions (e.g. inert gas
environments) are common in the art and will be applied when applicable.
Exemplary methods of preparing the compounds of the invention are
shown in the Schemes below.
General aspects of these exemplary methods are described below. Each
of the products of the following processes is optionally separated, isolated,
and/or purified prior to its use in subsequent processes.
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 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., with compound two.
"Treating" indicates the reasonable and usual manner in which organic
chemicals are allowed to react. Normal concentrations (0.01M to 10M,
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typically 0.1M to 1M), temperatures (-100 C to 250 C, typically -78 C to 150
C,
more typically -78 C to 100 C, still more typically 0 C to 100 C), reaction
vessels
(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 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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Scheme 1
O H~N ,,\CO2H 0 H~ CO2CH3
--- ~
H3C~N "'OH H3CN
H OH H OH
2
CO2CH3
O R 11,1 N~
--- ~ -.~~
H3C N =
H OH
3
O Rl~ N CO2CH3 O R' N CO2H
~ I I
H3C N = H3C N
H NH2 H NH2
4 5
In one embodiment, compounds of the invention are prepared as
depicted in Scheme 1. Siastatin B (1) from natural materials (Umezawa, H. et
al. "J. Antibiotics", 27:963-969 (1974) or ribose (Nishimura, Y. et al. "J.
Am.
Chem. Soc.", 110:7249-7250, 1988); and "Bull. Chem. Soc. Jpn.", 65:978-986,
1992)
is available in either enantiomer. Conversion to compound 2 is accomplished
by known methods (Nishimura, Y. et al. "J. Antibiotics", 46(2):300-309, 1993).
Reductive alkylation to form 3 is accomplished by known methods
(Nishimura, Y. et al. "J. Antibiotics", 45(10):1662-1668, 1992). Conversion of
the
alcohol 3 to the amine 4 is accomplished by the methods of Zbiral, E. et al.
"Liebigs Ann. Chem.", 129-134 (1991); and von Itzstein, M. et al.
"Carbohydrate
Res.", 244:181-185 (1993) Deprotection provides compound 5.
By way of example and not limitation, compounds 5 wherein Rl is
ethyl (Et, -CH2CH3), 1-propyl (n-Pr, n-propyl, -CH2CH2CH3), 1-butyl (n-Bu, n-
butyl, -CH2CH2CH2CH3), 2-methyl-l-propyl (i-Bu, i-butyl, -CH2CH(CH3)2), 1-
pentyl (n-pentyl, -CH2CH2CH2CH2CH3), 3-methyl-l-butyl (-
CH2CH2CH(CH3)2),
2-methyl-l-butyl (-CH2CH(CH3)CH2CH3), 1-hexyl
(-
CH2CH2CH2CH2CH2CH3), 2-ethyl-l-butyl (-CH2CH(CH2CH3)2), 2-ethyl-4-
phenyl-l-butyl
(-CH2CH(CH2CH3)(CH2CH2Ph)), or 2-(2-phenylethyl)-4-phenyl-l-butyl
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(-CH2CH(CH2CH2Ph)2) are prepared by the method of Scheme 1.
Scheme 2
O H, N CO2H O Pgl N CO2CH3
-- ~
H3C'J~ N'\" OH H3C
H OH H OH
6 7
Pg= N CO2CH3
N'\" ~ ----
H3C 0
H NH2
8
O Pg\ N CO2CH3 O H "N C02H
I I
---
H3C'KN"" H3C N'""
H NR2R3 H NR2R3
g 10a
III
R2R3(y,,, C02H
0 H3CN N
i
H H
10b
In another embodiment, compounds of the invention are prepared as
depicted in Scheme 2. The enantiomer of Siastatin B (6) is prepared from
ribose (Nishimura, Y. et al., "J. Am. Chem. Soc.", 110:7249-7250, 1988); and
"Bull. Chem. Soc. Jpn.", 65:978-986, 1992). Conversion to protected compound
7 is accomplished by known methods (e.g. Pg is Boc, Nishimura, Y. et al. "J.
Antibiotics", 46(2):300-309, 1993). Conversion of the alcohol 7 to the amine 8
is
accomplished by the methods of Zbiral, E. et al. "Liebigs Ann. Chem.", 129-
134,
1991; and von Itzstein, M. et al. "Carbohydrate Res.", 244:181-185, 1993.
Reductive alkylation to form 9 is accomplished by known methods
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(Nishimura, Y. et al. "J. Antibiotics", 45(10):1662-1668, 1992). Deprotection
provides compound 10a. Compound l0a is compound 10b
By way of example and not limitation, compounds 10b wherein R2 is H
and R3 is ethyl (Et, -CH2CH3), 1-propyl (n-Pr, n-propyl, -CH2CH2CH3), 1-butyl
(n-Bu, n-butyl, -CH2CH2CH2CH3), 2-methyl-l-propyl (i-Bu, i-butyl,
-CH2CH(CH3)2), 1-pentyl (n-pentyl, -CH2CH2CH2CH2CH3), 3-methyl-l-butyl
(-CH2CH2CH(CH3)2), 2-methyl-l-butyl (-CH2CH(CH3)CH2CH3), 1-hexyl
(-CH2CH2CH2CH2CH2CH3), 2-ethyl-l-butyl (-CH2CH(CH2CH3)2), 2-ethyl-4-
phenyl-l-butyl (-CH2CH(CH2CH3)(CH2CH2Ph)), or 2-(2-phenylethyl)-4-
phenyl-l-butyl (-CH2CH(CH2CH2Ph)2) are prepared by the method of Scheme
2.
Scheme 3
0 H~N \C02H OPg CO2CH3
--- a
H3C'~Tl D*IIVOH H3C'j,
N
i
H OH H OH
1 11
O Pg~NaC02CH3
---- ~ H3C N
H NR2R3
12
0 H~N CO2H
H3C'J~ N
H NR2R3
13
In another embodiment, compounds of the invention are prepared as
depicted in Scheme 3. Siastatin B (1) from natural materials (Umezawa, H. et
al., "J. Antibiotics", 27:963-969, 1994) or ribose (Nishimura, Y. et al., "J.
Am.
, 20 Chem. Soc.", 110:7249-7250, 1988); and "Bull. Chem. Soc. jpn.", 65:978-
986, 1992)
is available in either enantiomer. Conversion to protected compound 11 is
accomplished by known methods (e.g. Pg is Boc, Nishimura, Y. et al., "J.
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Antibiotics", 46(2):300-309, 1993). Conversion of the alcohol 11 to the amine
is
accomplished by the methods of Zbiral, E. et al., "Liebigs Ann. Chem.", 129-
134,
1991); and von Itzstein, M. et al., "Carbohydrate Res.", 244:181-185, 1993 and
reductive alkylation to form 12 is accomplished by known methods
(Nishimura, Y. et al. "J. Antibiotics", 45(10):1662-1668, 1992). Deprotection
provides compound 13.
By way of example and not limitation, compounds 13 wherein R2 is H
and R3 is ethyl (Et, -CH2CH3), 1-propyl (n-Pr, n-propyl, -CH2CH2CH3), 1-butyl
(n-Bu, n-butyl, -CH2CH2CH2CH3), 2-methyl-l-propyl (i-Bu, i-butyl,
-CH2CH(CH3)2), 1-pentyl (n-pentyl, -CH2CH2CH2CH2CH3), 3-methyl-l-butyl
(-CH2CH2CH(CH3)2), 2-methyl-l-butyl (-CH2CH(CH3)CH2CH3), 1-hexyl
(-CH2CH2CH2CH2CH2CH3), 2-ethyl-l-butyl (-CH2CH(CH2CH3)2), 2-ethyl-4-
phenyl-l-butyl (-CH2CH(CH2CH3)(CH2CH2Ph)), or 2-(2-phenylethyl)-4-
phenyl-l-butyl (-CH2CH(CH2CH2Ph)2) are prepared by the method of Scheme
3.
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Scheme 4
TrO-"~=,,; -
O TrO TrO
OH OTBDPS
100 101 102
HO
-- -~ -->-
TrO O TrO
OTBDPS OTBDPS
103 104
N3 CbzHN CbzHN
,. -
Tr0 TrO HO
OTBDPS OTBDPS OTBDPS
105 106 107
CbzN CbzN
-~ --,-
PthN AcHN
OTBDPS OTBDPS
108 109
O'\O O/ O
CbzN N
AcHN AcHN
OTBDPS OTBDPS
110 111
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Scheme 5
O" O HO OH
N ---~ --~
AcHN - AcHN -
OTBDPS OTBDPS
111 112
'TN
AcHN
OTBDPS
113
N OH '~N I OH
+
AcHN OTBDPS AcHN OTBDPS
114 115
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Scheme 6
O
OH H
-~ --
AcHN = AcHN =
OTBDPS OTBDPS
114 116
O O
OCH3 N OCH3
AcHN _ AcHN - ~
OTBDPS OH
117 118
O
N OH
AcHN =
OH
119
O
N I OH N I OCH3
--~
AcHN AcHN
OTBDPS
OTBDPS
115 0 120
OH
AcHN =
OH
121
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Scheme 7
HO OH HO O
N H
AcHN AcHN
OTBDPS OTBDPS
112 122
HO O
N OCH3 ~
AcHN
OTBDPS
123
O O
)JJ-)OCH3 + N OCH3
AcHN - AcHN
OTBDPS OTBDPS
117 120
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Scheme 4 is described in the Examples section below.
Scheme 5 and 6 are described below:
Acetonide 111 is converted to the diol 112 by treatment with acid catalyst
in methanol as described in "Protective Groups in Organic Synthesis" 2nd ed.,
T. W. Greene and P. G. M. Wuts, John Wiley & Sons, New York, NY, 1991.
Diol 112 upon treatment with p-toluenesulfonyl chloride in pyridine
solvent affords the primary tosylate which is converted to epoxide 113 with
potassium carbonate in methanol. An example of such a transformation is
described in "J. Org. Chem." 57:86 (1992).
Epoxide 113 is transformed to allylic alcohols 114 and 115 by treatment
with base. Isomerizations of epoxides to allylic alcohols are described in
"Org.
React." 29:345 (1983). The allylic alcohols are then separated by standard
chromatographic techniques. Alternatively, the synthesis of allylic alcohols
is
affected by sequential reaction with trimethylsilyl triflate and DBU as
described
in "J. Am. Chem. Soc." 101:2738 (1979).
Allylic alcohol 114 is oxidized to the corresponding aldehyde 116 using
MnO2. Such oxidations are described in "Synthesis", 601 (1986). This
oxidation is also accomplished with pyridine=S03 complex/DMSO/Et3N. An
example of such a transformation is described in "Synthesis", 274 (1988).
a, (3-Unsaturated aldehyde 116 is oxidized to the carboxylic acid methyl
ester 117 by treatment with sodium cyanide, Mn02 and acetic acid in methanol
solvent as described in "J. Am. Chem. Soc." 90:5616 (1968).
Deprotection of silyl ether 117 to alcohol 118 is carried out using
tetrabutylammonium fluoride. Hydrolysis of the methyl ester to carboxylic
acid 119 is conducted with potassium hydroxide. Both of these deprotection
methods are described in "Protective Groups in Organic Synthesis" 2nd ed., T.
W. Greene and P. G. M. Wuts, John Wiley & Sons, New York, NY, 1991.
In a similar fashion as described for 114, allylic alcohol 115 is converted
to carboxylic acid 121.
Diol 112 is oxidized with pyridine=SO3 complex/DMSO/Et3N to afford
a-hydroxyaldehyde 122 as described in "J. Chem. Soc. Chem. Commun." 18:
2197 (1994). The same reference describes the oxidation of an a-
hydroxyaldehyde to the corresponding a-hydroxycarboxylic acid using
NaC1O2/NaH2PO4/2-methyl-2-butene. Treatment of the carboxylic acid with
diazomethane affords carboxylic ester 123. Such an esterification of a
carboxylic acid is described in "Tetrahedron Lett." 1397 (1973).
a-Hydroxyester 123 is dehydrated to a mixture unsaturated esters 117
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and 120 with SOC12 and pyridine. Example of a similar reaction is described in
"J. Org. Chem." 60:2753 (1995).
Hydroxyesters 117 and 120 are deprotected to 119 and 121 respectively
using conditions previously described.
Modification of the exemplary starting materials to form different El
groups has been described in detail and will not be elaborated here. See
Fleet,
G.W.J. et al., "J. Chem. Soc. Perkin Trans. I", 905-908 (1984), Fleet, G.W.J.
et al.,
"J. Chem. Soc., Chem. Commun.", 849-850 (1983), Yee, Ying K. et al., "J. Med.
Chem.", 33:2437-2451 (1990); Olson, R.E. et al., "Bioorganic & Medicinal
Chemistry Letters", 4(18):2229-2234 (1994); Santella, J.B. III et al.,
"Bioorganic &
Medicinal Chemistry Letters", 4(18):2235-2240 (1994); Judd, D.B. et al., "J.
Med.
Chem.", 37:3108-3120 (1994) and De Lombaert, S. et al., "Bioorganic &
Medicinal Chemistry Letters", 5(2):151-154 (1994).
The El sulfur analogs of the carboxylic acid compounds of the
invention are prepared by any of the standard techniques. By way of example
and not limitation, the carboxylic acids are reduced to the alcohols by
standard
methods. The alcohols are converted to halides or sulfonic acid esters by
standard methods and the resulting compounds are reacted with NaSH to
produce the sulfide product. Such reactions are described in Patai, "The
Chemistry of the Thiol Group" (John Wiley, New York, 1974), pt. 2, and in
particular pages 721-735.
Modifications of each of the above schemes leads to various analogs of
the specific exemplary materials produced above. The above cited citations
describing suitable methods of organic synthesis are applicable to such
modifications.
In each of the above exemplary schemes it may be advantageous to
separate 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. Chromatography can involve any number of methods
including, for example, size exclusion or ion exchange chromatography, high,
medium, or low pressure liquid chromatography, small scale and preparative
thin or thick layer chromatography, as well as techniques of small scale thin
layer and flash chromatography.
Another class of separation methods involves treatment of a mixture
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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 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 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 extraction, and the like. One skilled in the art will apply
techniques most likely to achieve the desired separation.
Examples
The following Examples refer to the Schemes.
Example 1: Alcohol 101: To magnesium turnings (0.90 g, 37.2 mmol) in THF
(50 mL) was added several drops of 1,2-dibromoethane as initiator followed by
the addition of 2-bromopropene (4.5 g, 37.2 mmol) over a period of 1 h.
Additional 2-bromopropene (0.68 g, 5.6 mmol) was added and stirred for 1.5 h.
To a slurry of Cul (0.7 g, 3.7 mmol) in THF (30 mL) at -30 C was added via
cannula the grignard reagent over 15 min and was stirred an additional 20
min at -30 C. A solution of epoxide 100 (9.4 g, 29.8 mmol) in THF (40 mL) was
then added to the grignard/CuI mixture and was stirred at -30 C for 1 h. The
reaction was quenched at 0 C with saturated NH4C1 (100 mL) followed by the
addition of 1N NH4OH to dissolve precipitated solids. Brine was added and
the product was extracted into ether. The organic phase was washed with
brine, dried (MgSO4), filtered and solvent was evaporated to afford alcohol
101
(11.8 g) which was suitable for further transformations: 'H NMR (CDC13) S
7.51-7.27 (m, 15H), 4.84 (s, 1H), 4.77 (d, 1H, J= 0.9), 3.97 (m, 1H), 3.20 (m,
2H),
2.30 (m, 1H), 2.21 (d, 2H, J= 6.6), 1.76 (s, 3H).
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Example 2: Silyl ether 102: To a solution of alcohol 101 (1.77 g, 4.9 mmol)
and
imidazole (1.5 g, 22 mmol) in DMF (5 mL) was added tert-butyldiphenylsilyl
chloride (2.0 g, 7.4 mmol). After stirring for 1 h at room temperature the
reaction mixture was diluted with water and extracted with several portions of
ether. The combined organic extracts were washed with water and brine and
were dried (MgSO4), filtered and evaporated. The residue was
chromatographed on silica gel (20/1-hexane/ethyl acetate) to afford silyl
ether
102 (2.85 g, 97%) as an oil: 1H NMR (CDC13) S 7.73-7.65 (m, 4H), 7.49-7.20 (m,
21H), 4.56 (s,1H), 4.50 (s, 1H), 4.02-3.98 (m, 1H), 3.11 (m, 2H), 2.45 (dd,
1H, J= 7.5,
13.5), 2.10 (dd, 1H, J= 5.4,13.5),1.35 (s, 3H), 1.09 (s, 9H).
Example 3: Epoxide 103: To a solution of 102 (44.3 g, 74.2 mmol) in CH2C12
(600 mL) at 0 C was added NaHCO3 (31 g, 371 mmol) and MCPBA (32 g, 50-60%
MCPBA w/w). After stirring for 1 h at 0 C, water (100 mL) was added and the
mixture was stirred for 1 h more. After evaporation of the CH2C12, the
reaction was diluted with water and extracted with ether. The organic phase
was washed with cold 0.5 M sodium thiosulfate, saturated NaHCO3, brine and
was dried (MgSO4), filtered and evaporated. The residue was chromatographed
on silica gel (2/1-CH2CI2/hexane) to afford epoxide 103 (35.2 g) as a 1.5:1
mixture of diastereomers.
Example 4: Allylic alcohol 104: To a solution of lithium diethylamide
(generated from 11.5 mL diethylamine and 52 mL of 2.1 M butyl lithium in
hexane) in ether (400 mL) at 0 C was added epoxide 103 (33.5 g) in ether (150
mL). The reaction was allowed to warm to room temperature and was stirred
for 17 h. The reaction was cooled to 0 C and was poured into ice-water
mixture (500 mL). The organic phase was washed with water, brine, and was
dried (MgSO4), filtered and evaporated. The crude product was
chromatographed on silica gel (3/1-hexane/ethyl acetate) to afford allylic
alcohol 104 (15.4 g, 46%) as an oil: 1H NMR (CDC13) S 7.75-7.65 (m, 4H), 7.53-
7.22 (m, 21H), 4.90 (m, 1H), 4.71 (s,1H), 4.03 (m, 1H), 3.73 (d, 2H, J= 5.1),
3.16 (d,
2H,J=5.1),2.53(dd,1H,J=6.9,13.8),2.25(dd,1H,J=5.1,13.8),1.54(brt,1H,J=
6), 1.12 (s, 9H).
Example 5: Allylic azide 105: To a solution of alcohol 104 (13.1 g, 21.4 mmol)
in CH202 (100 mL) at 0 C was added triethylamine (7.45 mL, 53.5 mmol)
followed by methanesulfonyl chloride (1.82 g, 23.5 mmol). After 30 min at 0 C
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cold water was added and CH2C12 was evaporated and the residue was
partitioned between ether and water. The organic phase was washed with
water, brine, and was dried (MgSO4), filtered, and evaporated. The crude
mesylate was dissolved in DMF (100 mL), sodium azide (2.8 g, 42.8 mmol) was
added and the mixture was stirred at room temperature for 30 min. Volatiles
were evaporated and the residue was partitioned between ether and water.
The organic phase was washed with water, brine and was dried (MgSO4),
filtered and evaporated. The crude product was chromatographed on silica gel
(10/1-hexane/ethyl acetate) to afford allylic azide 105 (11.9 g, 87%) as an
oil: 1H
NMR (CDC13) S 7.71-7.60 (m, 4H), 7.51-7.21 (m, 21H), 4.89 (d, 1H, J= 1.5),
4.78 (s,
1H), 3.97 (m, 1H), 3.36 (s, 2H), 3.10 (d, 2H, J= 5.1), 2.56 (dd, 1H, J= 6.3,
13.8), 2.20
(dd,1H, J = 5.1, 13.8), 1.08 (s, 9H).
Example 6: Carbamate 106: To a solution of azide 105 (9.3 g, 14.6 mmol) in
CH3CN (100 mL) and water (5 mL) at 0 C was added trimethylphosphine (2.3
mL, 22 mmol). After stirring for 2 h at 0 C the reaction was warmed to room
temperature and was stirred for 18 h. To the solution was added potassium
carbonate (4.0 g, 29.2 mmol) and water (20 mL) followed by addition of benzyl
chloroformate (3.1 mL, 95% purity, -21 mmol). The reaction was stirred for 1.5
h and solvent was evaporated. Water was added and the organics were
extracted into ether. The combined organic extracts were washed with brine,
dried (MgSO4), filtered, and evaporated. The residue was chromatographed on
silica gel (3/1-hexane ethyl acetate) to afford carbamate 106 (10.6 g, 97%) as
a
thick oil: 'H NMR (CDC13) S 7.72-7.60 (m, 4H), 7.51-7.20 (m, 26H), 5.08 (br s,
2H), 4.76 (s, 1H), 4.66 (s, 1H), 4.50 (m, 1H), 3.99 (m, 1H), 3.45 (dd, 1H, J=
5.7, 16),
3.33(dd,1H,J=5.7,16),3.12(d,2H),2.48(dd,1H,J=6.6,13.8),2.16(dd,1H,J=6,
13.8),1.08 (s, 9H).
Example 7: Alcohol 107: To a solution of 106 (10.6 g, 14.2 mmol) in CH2C12 (20
mL). and methanol (20 mL) at 0 C was added formic acid (25 mL). The solution
was stirred at 0 C for 2.5 h and was then poured into saturated NaHCO3 (600
mL) with stirring. The aqueous phase was saturated with NaCI and extracted
with ethyl acetate. The combined organic extracts were dried (MgSO4),
filtered,
and evaporated. The crude residue was chromatographed on silica gel (2/1-
hexane ethyl acetate) to afford alcohol 107 (5.9 g, 83%) as an oil: 1H NMR
(CDC13) S 7.73-7.68 (m, 4H), 7.50-7.34 (m, 11H), 5.09 (s, 2H), 4.92 (d, 1H, J=
1.2),
4.79 (s, 1H), 4.68 (m, 1H), 3.92 (m, 1H), 3.60-3.39 (m, 4H), 2.36 (dd, 1H, J=
8.4,
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13.5), 2.15 (dd,1H J= 4.8, 13.5), 1.94 (br t,1H)1.10 (s, 9H).
Example 8: Phthalimide adduct 108: To a solution of DMSO (0.76 mL, 10.8
mmol) in CH202 (4 mL) at -78 C was added oxalyl chloride (0.39 mL, 4.5
mmol). After stirring for 5 min a solution of alcohol 107 (2.19 g, 4.3 mmol)
in
CH2C12 (5 mL) was added, stirred 25 min, Triethylamine (3 mL, 21.5 mmol)
was added, the reaction was stirred at -78 C for 30 min and was warmed to
room temperature. After 1 h the reaction was diluted with ether, filtered, and
evaporated. The residue was dissolved in ethyl acetate, washed with water,
brine and the organic phase was dried (MgSO4), filtered, and evaporated. To a
solution of the residue in DMF (20 mL) was added Ph3P (2.2 g, 8.3 mmol) and
phthalimide (1.22 g, 8.3 mmol) and the solution was cooled to 0 C at which
time diethylazodicarboxylate (1.3 mL, 8.3 mmol) was added. The reaction was
stirred at 0 C for 1 h and was warmed to room temperature stirring for 3.5 h.
Water was added to quench excess diethylazodicarboxylate and DMF was
evaporated. The residue was chromatographed on silica gel (2% ethyl acetate
in CH2C12). Rechromatography of the mixed fractions (1% ethyl acetate in
CH2C12) afforded phthalimide adduct 108 (1.95 g, 72%) as a foam. 1H NMR
(CDC13) S 7.83-7.08 (m, 19H), 5.98 (br d, 1H, J= 6.9), 5.22 (d, 1H, J= 12.3),
4.97 (d,
1H, J= 12.3), 4.88 (s,1H), 4.71 (s, 1H), 4.60-4.52 (m, 2H), 4.20 (br d, 1H, J=
16.2),
2.54 (dd, 1H, J= 4, 15.6), 2.41-2.33 (m, 1H), 0.98 (s, 9H).
Example 9: Acetamide 109: A solution of 108 (4.2 g, 6.7 mmol) in MeOH (100
mL) was treated with hydrazine monohydrate (1.6 mL, 33 mmol) at 40 C for 5
h. Solvent was evaporated and the residue was suspended in ethyl acetate and
the solid residue was removed by filtration. Evaporation of the filtrate gave
a
residue which was dissolved in pyridine (30 mL), and cooled to 0 C. Acetic
anhydride (6.9 mL, 73 mmol) was added and the reaction was warmed to room
temperature stirring for 2 h. After evaporation of the volatiles the residue
was
dissolved in ethyl acetate and washed with water and brine. The organic
phase was dried (MgSO4), filtered, evaporated and chromatographed on silica
gel (1/1 hexane-ethyl acetate) to afford 2.7 g of purified product.
Precipitation
from ether/hexane gave acetamide 109 (1.89 g, 52%) as a white solid. 'H NMR
(CD3OD) S 7.69-7.62 (m, 4H), 7.45-7.32 (m, 11H), 6.23 (d, 1H, j= 2.4), 5.22
(br d,
1H), 5.04 (br s, 2H), 4.76 (s, 1H), 4.56 (d,1H, J= 14.4), 4.03 (br s,1H), 3.73
(br d,
1H), 2.43 (br d, 1H), 2.10 (br d, 1H), 1.93 (s, 3H), 1.02 (s, 9H)
Example 10: Acetonide 110: To a solution of 109 (478 mg, 0.88 mmol) in
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acetone (3 mL) was added 4-methylmorpholine N-oxide (291 mg, 2.5 mmol)
and 0.04M Os04 solution in water (1 mL). The reaction was stirred for 16 h at
room temperature, cooled to 0 C and was treated with 10 sodium thiosulfate
solution (10 mL). The mixture was diluted with brine and extracted with ethyl
acetate. The combined organic extracts were dried (MgSO4), filtered,
evaporated and the residue was dissolved in ethyl acetate and filtered through
a pad of silica gel. Evaporation of the filtrate gave a residue which was
dissolved in acetone (10 mL) and was treated with dimethoxypropane (5 mL)
and a catalytic amount of p-toluenesulfonic acid. After stirring for 15 min at
room temperature saturated NaHCO3 was added and solvents were
evaporated. The product was extracted into ethyl acetate and the combined
organic extracts were dried (MgSO4), filtered, and evaporated. The residue
chromatographed on silica gel (1/1 hexane-ethyl acetate) to afford acetonide
110 (465 mg, 85%) as a white solid.1H NMR (CD3OD) 8 7.62 (br s, 4H), 7.51-7.23
(m, 11H), 6.05 (br s,1H), 5.20 (br d, 1H), 5.05 (br d,1H), 4.11 (m, 3H), 3.71
(br s,
1H), 2.85 (br d,1H), 2.04 (m,1H),1.91 (s, 3H), 1.62 (m, 1H),1.41 (s, 3H),1.18
(br s,
3H), 1.04 (s, 9H).
Example 11: N-alkyl derivative 111: A solution of 110 (465 mg, 0.75 mmol)
was treated with 10% Pd/C (90 mg) and was stirred under hydrogen (balloon)
for 4 h. The catalyst was removed by filtration (celite )*and the filtrate was
evaporated to afford a residue which was used directly. To a solution of the
amine (324 mg, 0.67 mmol) in methanol (4 mL) was added 2-
ethylbutyraldehyde (0.83 mL, 67 mmol) and the solution was cooled to 0 C. To
this solution was added 1.34 mL of NaCNBH3/ZnC12 reagent prepared from
NaCNBH3 (314 mg, 5 mmol) and ZnC12 (340 mg, 2.5 mmol) in methanol (10
mL). The reaction was stirred at 0 C for 40 min and was evaporated. The
residue was partitioned between ether and 0.1N NaOH. The organic phase
was dried (MgSO4), filtered and evaporated. The residue was
chromatographed on silica gel (2/1-hexane/ethyl acetate) to afford N-alkyl
derivative 111 (212 mg, 56%) as a white solid. An analytical sample was
recrystallized from hexane as thin needles: mp 136-138 C; 'H NMR (CDC13) S
7.76-7.66 (m, 4H), 7.50-7.36 (m, 6H), 5.58 (d, 1H, J= 9), 4.84 (dd,1H, J= 2.7,
9), 4.26
(d,1H,J=9),4.01 (d,1H,J=3.3),3.74(d,1H,J=8.7),2.61 (d,1H,J=11.7),2.37(dd,
1H,J=6,12.3),2.30(d,1H,J=12),2.20(dd,1H,J=5.7,12.6),1.92(s,3H),1.69(dd,
1H, J= 3.3,14.1),1.54-1.28 (m, 9H), 1.17 (s, 3H), 1.13 (s, 9H), 0.96-0.82 (m,
6H).
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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
claims. It is apparent that certain modifications of the methods and
compositions
of the following claims can be made within the scope and spirit of the
invention.
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