U.S. patent application number 12/504243 was filed with the patent office on 2010-05-20 for sulphonamide derivatives as prodrugs of aspartyl protease inhibitors.
This patent application is currently assigned to Vertex Pharmaceuticals Incorporated. Invention is credited to Christopher Todd Baker, Eric Steven Furfine, Michael Robin Hale, Istvan Kaldor, Wieslaw Mieczylaw Kazmierski, Andrew Spaltenstein, Roger Dennis Tung.
Application Number | 20100124543 12/504243 |
Document ID | / |
Family ID | 25544691 |
Filed Date | 2010-05-20 |
United States Patent
Application |
20100124543 |
Kind Code |
A1 |
Tung; Roger Dennis ; et
al. |
May 20, 2010 |
SULPHONAMIDE DERIVATIVES AS PRODRUGS OF ASPARTYL PROTEASE
INHIBITORS
Abstract
The present invention relates to prodrugs of a class of
sulfonamides which are aspartyl protease inhibitors. In one
embodiment, this invention relates to a novel class of prodrugs of
HIV aspartyl protease inhibitors characterized by favorable aqueous
solubility, high oral bioavailability and facile in vivo generation
of the active ingredient. This invention also relates to
pharmaceutical compositions comprising these prodrugs. The prodrugs
and pharmaceutical compositions of this invention are particularly
well suited for decreasing the pill burden and increasing patient
compliance. This invention also relates to methods of treating
mammals with these prodrugs and pharmaceutical compositions.
Inventors: |
Tung; Roger Dennis;
(Beverly, MA) ; Hale; Michael Robin; (Bedford,
MA) ; Baker; Christopher Todd; (Waltham, MA) ;
Furfine; Eric Steven; (Durham, NC) ; Kaldor;
Istvan; (Durham, NC) ; Kazmierski; Wieslaw
Mieczylaw; (Releigh, NC) ; Spaltenstein; Andrew;
(Raleigh, NC) |
Correspondence
Address: |
ROPES & GRAY LLP
PATENT DOCKETING 39/361, 1211 AVENUE OF THE AMERICAS
NEW YORK
NY
10036-8704
US
|
Assignee: |
Vertex Pharmaceuticals
Incorporated
Cambridge
MA
|
Family ID: |
25544691 |
Appl. No.: |
12/504243 |
Filed: |
July 16, 2009 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10958223 |
Oct 4, 2004 |
7592368 |
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12504243 |
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10370171 |
Feb 19, 2003 |
6838474 |
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10958223 |
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09602494 |
Jun 23, 2000 |
6559137 |
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10370171 |
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PCT/US98/04595 |
Mar 9, 1998 |
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09602494 |
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08998050 |
Dec 24, 1997 |
6436989 |
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PCT/US98/04595 |
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Current U.S.
Class: |
424/85.2 ;
424/85.1; 424/85.7; 514/220; 514/254.1; 514/263.38; 514/336;
514/397; 514/422; 514/43; 514/45; 514/471; 514/487; 514/49; 514/50;
514/512; 514/54; 544/374; 546/284.4; 548/315.7; 548/517; 549/475;
558/275; 560/13 |
Current CPC
Class: |
C07D 307/20 20130101;
C07H 15/04 20130101; C07D 405/12 20130101; C07F 9/65844 20130101;
C07F 9/65515 20130101; A61P 43/00 20180101; C07C 311/18 20130101;
A61P 31/12 20180101; A61P 31/18 20180101 |
Class at
Publication: |
424/85.2 ;
549/475; 544/374; 548/517; 546/284.4; 548/315.7; 558/275; 560/13;
514/471; 514/254.1; 514/422; 514/336; 514/397; 514/487; 514/512;
514/50; 514/49; 514/45; 514/54; 514/43; 514/263.38; 514/220;
424/85.1; 424/85.7 |
International
Class: |
A61K 31/4025 20060101
A61K031/4025; C07D 307/20 20060101 C07D307/20; C07D 405/12 20060101
C07D405/12; C07C 69/96 20060101 C07C069/96; C07C 271/20 20060101
C07C271/20; A61P 31/12 20060101 A61P031/12; A61K 31/341 20060101
A61K031/341; A61K 31/496 20060101 A61K031/496; A61K 31/443 20060101
A61K031/443; A61K 31/4178 20060101 A61K031/4178; A61K 31/27
20060101 A61K031/27; A61K 31/265 20060101 A61K031/265; A61K 31/7072
20060101 A61K031/7072; A61K 31/7068 20060101 A61K031/7068; A61K
31/708 20060101 A61K031/708; A61K 31/715 20060101 A61K031/715; A61K
31/52 20060101 A61K031/52; A61K 31/551 20060101 A61K031/551; A61K
38/20 20060101 A61K038/20; A61K 38/19 20060101 A61K038/19; A61K
38/21 20060101 A61K038/21 |
Claims
1. A compound of formula I: ##STR00154## wherein: A is selected
from Ht; --R.sup.1--Ht; each R.sup.1 is independently selected from
--C(O)--, --S(O).sub.2--, --C(O)--C(O)--, --O--C(O)--,
--O--S(O).sub.2, --N(R.sup.2)--S(O).sub.2--, --N(R.sup.2)--C(O)--
or --N(R.sup.2)--C(O)--C(O)--; each Ht is independently selected
from C.sub.3-C.sub.7 cycloalkyl; C.sub.5-C.sub.7 cycloalkenyl;
C.sub.6-C.sub.10 aryl; or a 5-7 membered saturated or unsaturated
heterocycle, containing one or more heteroatoms selected from N,
N(R.sup.2), O, S and S(O).sub.n; wherein said aryl or said
heterocycle is optionally fused to Q; and wherein any member of
said Ht is optionally substituted with one or more substituents
independently selected from oxo, --OR.sup.2, SR.sup.2, --R.sup.2,
--N(R.sup.2).sub.2, --R.sup.2--OH, --CN, --C(O)O--R.sup.2,
--C(O)--N(R.sup.2).sub.2, --S(O).sub.2--N(R.sup.2).sub.2,
--N(R.sup.2)--C(O)--R.sup.2, --C(O)--R.sup.2,
--S(O).sub.n--R.sup.2, --OCF.sub.3, --S(O).sub.n-Q, methylenedioxy,
--N(R.sup.2)--S(O).sub.2--R.sup.2, halo, --CF.sub.3, --NO.sub.2, Q,
--OQ, --OR.sup.7, --SR.sup.7, --R.sup.7, --N(R.sup.2)(R.sup.7) or
--N(R.sup.7).sub.2; each R.sup.2 is independently selected from H,
or (C.sub.1-C.sub.4)-alkyl optionally substituted with Q; B, when
present, is --N(R.sup.2)--C(R.sup.3).sub.2--C(O)--; each x is
independently 0 or 1; each R.sup.3 is independently selected from
H, Ht, (C.sub.1-C.sub.6)-alkyl, (C.sub.2-C.sub.6)-alkenyl,
(C.sub.3-C.sub.6)-cycloalkyl or (C.sub.5-C.sub.6)-cycloalkenyl;
wherein any member of said R.sup.3, except H, is optionally
substituted with one or more substituents selected from --OR.sup.2,
--C(O)--NH--R.sup.2, --S(O).sub.n--N(R.sup.2).sub.2, Ht, --CN,
--SR.sup.2, --CO.sub.2R.sup.2, N(R.sup.2)--C(O)--R.sup.2; each n is
independently 1 or 2; G, when present, is selected from H, R.sup.7
or (C.sub.1-C.sub.4)-alkyl, or, when G is (C.sub.1-C.sub.4)-alkyl,
G and R.sup.7 are bound to one another either directly or through a
C.sub.1-C.sub.3 linker to form a heterocyclic ring; or when G is
absent, the atom to which G is attached is bound directly to the
R.sup.7 group in --OR.sup.7 with the concomitant displacement of
one -ZM group from R.sup.7; D and D' are independently selected
from Q; (C.sub.1-C.sub.6)-alkyl, which is optionally substituted
with one or more groups selected from (C.sub.3-C.sub.6)-cycloalkyl,
--OR.sup.2, --R.sup.3, --O-Q or Q; (C.sub.2-C.sub.4)-alkenyl, which
is optionally substituted with one or more groups selected from
(C.sub.3-C.sub.6)-cycloalkyl, --OR.sup.2, --R.sup.3, --O-Q or Q;
(C.sub.3-C.sub.6)-cycloalkyl, which is optionally substituted with
or fused to Q; or (C.sub.5-C.sub.6)-cycloalkenyl, which is
optionally substituted with or fused to Q; each Q is independently
selected from a 3-7 membered saturated, partially saturated or
unsaturated carbocyclic ring system; or a 5-7 membered saturated,
partially saturated or unsaturated heterocyclic ring containing one
or more heteroatoms selected from O, N, S, S(O).sub.n or
N(R.sup.2); wherein any ring in Q is optionally substituted with
one or more groups selected from oxo, --OR.sup.2, --R.sup.2,
--N(R.sup.2).sub.2, --N(R.sup.2)--C(O)--R.sup.2, --R.sup.2--OH,
--CN, --C(O)OR.sup.2, --C(O)--N(R.sup.2).sub.2, halo or --CF.sub.3;
E is selected from --O--R.sup.3; --N(R.sup.2)(R.sup.3);
(C.sub.1-C.sub.6)-alkyl, which is optionally substituted with one
or more groups selected from R.sup.4 or Ht;
(C.sub.2-C.sub.6)-alkenyl, which is optionally substituted with one
or more groups selected from R.sup.4 or Ht; each R.sup.4 is
independently selected from --OR.sup.2, --SR.sup.2,
--C(O)--NHR.sup.2, --S(O).sub.2--NHR.sup.2, halo,
--N(R.sup.2)--C(O)--R.sup.2, --N(R.sup.2).sub.2 or --CN; each
R.sup.7 is independently selected from ##STR00155## wherein each M
is independently selected from H, Li, Na, K, Mg, Ca, Ba,
--N(R.sup.2).sub.4, (C.sub.1-C.sub.12)-alkyl,
(C.sub.2-C.sub.12)-alkenyl, or --R.sup.6; wherein 1 to 4 --CH.sub.2
radicals of the alkyl or alkenyl group, other than the --CH.sub.2
that is bound to Z, is optionally replaced by a heteroatom group
selected from O, S, S(O), S(O).sub.2, or N(R.sup.2); and wherein
any hydrogen in said alkyl, alkenyl or R.sup.6 is optionally
replaced with a substituent selected from oxo, --OR.sup.2,
--R.sup.2, N(R.sup.2).sub.2, N(R.sup.2).sub.3, R.sup.2OH, --CN,
--C(O)OR.sup.2, --C(O)--N(R.sup.2).sub.2,
S(O).sub.2--N(R.sup.2).sub.2, N(R.sup.2)--C(O)--R.sub.2,
C(O)R.sup.2, --S(O).sub.n--R.sup.2, OCF.sub.3,
--S(O).sub.n--R.sup.6, N(R.sup.2)--S(O).sub.2--R.sup.2, halo,
--CF.sub.3, or --NO.sub.2; M' is H, (C.sub.1-C.sub.12)-alkyl,
(C.sub.2-C.sub.12)-alkenyl, or --R.sup.6; wherein 1 to 4 --CH.sub.2
radicals of the alkyl or alkenyl group is optionally replaced by a
heteroatom group selected from O, S, S(O), S(O).sub.2, or
N(R.sup.2); and wherein any hydrogen in said alkyl, alkenyl or
R.sup.6 is optionally replaced with a substituent selected from
oxo, --OR.sup.2, --R.sup.2, --N(R.sup.2).sub.2, N(R.sup.2).sub.3,
--R.sup.2OH, --CN, --CO.sub.2R.sup.2, --C(O)--N(R.sup.2).sub.2,
--S(O).sub.2--N(R.sup.2).sub.2, --N(R.sup.2)--C(O)--R.sup.2,
--C(O)R.sup.2, --S(O).sub.n--R.sup.2, --OCF.sub.3,
--S(O).sub.n--R.sup.6, --N(R.sup.2)--S(O).sub.2--R.sup.2, halo,
--CF.sub.3, or --NO.sub.2; Z is CH.sub.3, O, S, N(R.sup.2).sub.2,
or, when M is not present, H. Y is P or S; X is O or S; and R.sup.9
is C(R.sup.2).sub.2, O or N(R.sup.2); and wherein when Y is S, Z is
not S; and R.sup.6 is a 5-6 membered saturated, partially saturated
or unsaturated carbocyclic or heterocyclic ring system, or an 8-10
membered saturated, partially saturated or unsaturated bicyclic
ring system; wherein any of said heterocyclic ring systems contains
one or more heteroatoms selected from O, N, S, S(O).sub.n or
N(R.sup.2); and wherein any of said ring systems optionally
contains 1 to 4 substituents independently selected from OH,
C.sub.1-C.sub.4 alkyl, O--(C.sub.1-C.sub.4)-alkyl or
O--C(O)--(C.sub.1-C.sub.4)-alkyl.
2. The compound according to claim 1, wherein at least one R.sup.7
is selected from: ##STR00156## ##STR00157##
3. The compound according to claim 2, wherein said compound has
formula XXII: ##STR00158## wherein A, D', R.sup.7 and E are as
defined in claim 1.
4-12. (canceled)
13. The compound according to claim 2, wherein said compound has
formula XXXI: ##STR00159##
14. The compound according to claim 13, wherein: A is R.sup.1-Ht;
each R.sup.3 is independently (C.sub.1-C.sub.6)-alkyl which is
optionally substituted with --OR.sup.2, --C(O)--NH--R.sup.2,
--S(O).sub.nN(R.sup.2).sub.2, --Ht, --CN, --SR.sup.2,
--CO.sub.2R.sup.2 or --N(R.sup.2)--C(O)--R.sup.2; and D' is
(C.sub.1-C.sub.4)-alkyl, which is optionally substituted with
(C.sub.3-C.sub.6)-cycloalkyl, --OR.sup.2, --O-Q; and E is
--N(R.sup.2)(R.sup.3).
15. The compound according to claim 14, wherein R.sup.7 in the
--OR.sup.7 group depicted in formula XXXI is --PO(OM).sub.2 or
--C(O)-M'.
16. (canceled)
17. A compound selected from: ##STR00160## ##STR00161## wherein
R.sup.10 is selected from isopropoyl or cyclopentyl; R.sup.11 is
selected from NHR.sup.7 or OR.sup.7; in compound 1005, when R.sup.7
is PO.sub.3M, (G).sub.x is not H; and x, R.sup.7 and G are as
defined in claim 1.
18. A pharmaceutical composition, comprising a compound according
to any one of claims 1 in an amount effective to treat infection by
a virus that is characterized by an aspartyl protease; and a
pharmaceutically acceptable carrier, adjuvant or vehicle.
19. The pharmaceutical composition according to claim 18, wherein
said virus is HIV.
20. The pharmaceutical composition according to claim 18, wherein
said pharmaceutical composition is formulated for oral
administration.
21. The pharmaceutical composition according to claim 18, further
comprising one or more agents selected from an anti-viral agent, an
HIV protease inhibitor other than a compound according to claim 1,
and an immunostimulator.
22. The pharmaceutical composition according to claim 21, further
comprising one or more agents selected from zidovudine (AZT),
zalcitabine (ddC), didanosine (ddI), stavudine (d4T), 3TC, 935U83,
1592U89, 524W91, saquinavir (Ro 31-8959), L-735,524, SC-52151, ABT
538 (A84538), AG 1343, XM 412, XM 450, CGP 53,437, tucaresol,
polysulfated polysaccharides, ganciclovir, dideoxycytidine,
ribavirin, acyclovir, TIBO, nevirapine, IL-2, GM-CSF, interferon
alpha, or erythropoietin (EPO).
23. A method for inhibiting aspartyl protease activity in a mammal,
comprising the step of administering to said mammal a
pharmaceutical composition according to claim 18.
24. A method for treating HIV infection in a mammal comprising the
step of administering to said mammal a pharmaceutical composition
according to claim 18.
25. The method according to claim 24, wherein said mammal is
additionally administered one or more additional agents selected
from an anti-viral agent, an HIV protease inhibitor other than a
compound according to claim 1, and an immunostimulator either as a
part of a single dosage form with said pharmaceutical composition
or as a separate dosage form.
26. The method according to claim 25, wherein said additional agent
is selected from zidovudine (AZT), zalcitabine (ddC), didanosine
(ddI), stavudine (d4T), 3TC, 935U83, 1592U89, 524W91, saquinavir
(Ro 31-8959), L-735,524, SC-52151, ABT 538 (A84538), AG 1343, XM
412, XM 450, CGP 53,437, tucaresol, polysulfated polysaccharides,
ganciclovir, dideoxycytidine, ribavirin, acyclovir, TIBO,
nevirapine, IL-2, GM-CSF, interferon alpha, or erythropoietin
(EPO).
27. The method according to claim 24, wherein said step of
administering comprises oral administration.
Description
TECHNICAL FIELD OF THE INVENTION
[0001] The present invention relates to prodrugs of a class of
sulfonamides which are aspartyl protease inhibitors. In one
embodiment, this invention relates to a novel class of prodrugs of
HIV aspartyl protease inhibitors characterized by favorable aqueous
solubility, high oral bioavailability and facile in vivo generation
of the active ingredient. This invention also relates to
pharmaceutical compositions comprising these prodrugs. The prodrugs
and pharmaceutical compositions of this invention are particularly
well suited for decreasing the pill burden and increasing patient
compliance. This invention also relates to methods of treating
mammals with these prodrugs and pharmaceutical compositions.
BACKGROUND OF THE INVENTION
[0002] Aspartyl protease inhibitors are considered the most
effective current drug in the fight against HIV infection. These
inhibitors, however, require certain physicochemical properties in
order to achieve good potency against the enzyme. One of these
properties is high hydrophobicity. Unfortunately, this property
results in poor aqueous solubility and low oral
bioavailability.
[0003] U.S. Pat. No. 5,585,397 describes a class of sulfonamide
compounds that are inhibitors of the aspartyl protease enzyme.
These compounds illustrate the drawbacks concomitant to
pharmaceutical compositions comprising hydrophobic aspartyl
protease inhibitors. For example, VX-478
(4-amino-N-((2-syn,3S)-2-hydroxy-4-phenyl-2((S)-tetrahydrofuran-3-yl-oxyc-
arbonylamino)-butyl-N-isobutyl-benzenesulfonamide) is an aspartyl
protease inhibitor disclosed in the '397 patent. It has a
relatively low aqueous solubility. While the oral bioavailability
of this inhibitor in a "solution" formulation is excellent, the
dosage of VX-478 in this form is severely limited by the amount of
liquid present in the particular liquid dosage from, e.g.,
encapsulated into a soft gelatin capsule. A higher aqueous
solubility would increase drug load per unit dosage of VX-478.
[0004] Currently, the solution formulation of VX-478 produces an
upper limit of 150 mg of VX-478 in each capsule. Given a
therapeutic dose of 2400 mg/day of VX-478, this formulation would
require a patient to consume 16 capsules per day. Such a high pill
burden would likely result in poor patient compliance, thus
producing sub-optimal therapeutic benefit of the drug. The high
pill burden is also a deterrent to increasing the amount of the
drug administered per day to a patient. Another drawback of the
pill burden and the concomitant patient compliance problem is in
the treatment of children infected with HIV.
[0005] Furthermore, these "solution" formulations, such as the
mesylate formulation, are at a saturation solubility of VX-478.
This creates the real potential of having the drug crystallize out
of solution under various storage and/or shipping conditions. This,
in turn, would likely result in a loss of some of the oral
bioavailability achieved with VX-478.
[0006] One way of overcoming these problems is to develop a
standard solid dosage form, such as a tablet or a capsule or a
suspension form. Unfortunately, such solid dosage forms have much
lower oral bioavailability of the drug.
[0007] Thus, there is a need to improve the drug load per unit
dosage form for aspartyl protease inhibitors. Such an improved
dosage form would reduce the pill burden and increase patient
compliance. It would also provide for the possibility of increasing
the amounts of the drug administered per day to a patient.
SUMMARY OF THE INVENTION
[0008] The present invention provides novel prodrugs of a class of
sulfonamide compounds that are inhibitors of aspartyl protease, in
particular, HIV aspartyl protease. These prodrugs are characterized
by excellent aqueous solubility, increased bioavailability and are
readily metabolized into the active inhibitors in vivo. The present
invention also provides pharmaceutical compositions comprising
these prodrugs and methods of treating HIV infection in mammals
using these prodrugs and the pharmaceutical compositions
thereof.
[0009] These prodrugs can be used alone or in combination with
other therapeutic or prophylactic agents, such as anti-virals,
antibiotics, immunomodulators or vaccines, for the treatment or
prophylaxis of viral infection.
[0010] It is a principal object of this invention to provide a
novel class of prodrugs of sulfonamide compounds that are aspartyl
protease inhibitors, and particularly, HIV aspartyl protease
inhibitors. This novel class of sulfonamides is represented by
formula I:
##STR00001##
wherein:
[0011] A is selected from H; Ht; --R.sup.1--Ht;
--R.sup.1--C.sub.1-C.sub.6 alkyl, which is optionally substituted
with one or more groups independently selected from hydroxy,
C.sub.1-C.sub.4 alkoxy, Ht, --O-Ht,
--NR.sup.2--CO--N(R.sup.2).sub.2 or --CO--N(R.sup.2).sub.2;
--R.sup.1--C.sub.2-C.sub.6 alkenyl, which is optionally substituted
with one or more groups independently selected from hydroxy,
C.sub.1-C.sub.4 alkoxy, Ht, --O-Ht,
--NR.sup.2--CO--N(R.sup.2).sub.2 or --CO--N(R.sup.2).sub.2; or
R.sup.7;
[0012] each R.sup.1 is independently selected from --C(O)--,
--S(O).sub.2--, --C(O)--C(O)--, --O--C(O)--, --O--S(O).sub.2,
--NR.sup.2--S(O).sub.2--, --NR.sup.2--C(O)-- or
--NR.sup.2--C(O)--C(O)--;
[0013] each Ht is independently selected from C.sub.3-C.sub.7
cycloalkyl; C.sub.5-C.sub.7 cycloalkenyl; C.sub.6-C.sub.10 aryl; or
a 5-7 membered saturated or unsaturated heterocycle, containing one
or more heteroatoms selected from N, N(R.sup.2), O, S and
S(O).sub.n; wherein said aryl or said heterocycle is optionally
fused to Q; and wherein any member of said Ht is optionally
substituted with one or more substituents independently selected
from oxo, --OR.sup.2, SR.sup.2, --R.sup.2, --N(R.sup.2)(R.sup.2),
--R.sup.2--OH, --CN, --CO.sub.2R.sup.2, --C(O)--N(R.sup.2).sub.2,
--S(O).sub.2--N(R.sup.2).sub.2, --N(R.sup.2)--C(O)--R.sup.2,
--C(O)--R.sup.2, --S(O).sub.n--R.sup.2, --OCF.sub.3,
--S(O).sub.n-Q, methylenedioxy, --N(R.sup.2)--S(O).sub.2 (R.sup.2),
halo, --CF.sub.3, --NO.sub.2, Q, --OQ, --OR.sup.7, --SR.sup.7,
--R.sup.7, --N(R.sup.2)(R.sup.7) or --N(R.sup.7).sub.2;
[0014] each R.sup.2 is independently selected from H, or
C.sub.1-C.sub.4 alkyl optionally substituted with Q;
[0015] B, when present, is
--N(R.sup.2)--C(R.sup.3).sub.2--C(O)--;
[0016] each x is independently 0 or 1;
[0017] each R.sup.3 is independently selected from H, Ht,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.3-C.sub.6
cycloalkyl or C.sub.5-C.sub.6 cycloalkenyl; wherein any member of
said R.sup.3, except H, is optionally substituted with one or more
substituents selected from --OR.sup.2, --C(O)--NH--R.sup.2,
--S(O).sub.n--N(R.sup.2)(R.sup.2), Ht, --CN, --SR.sup.2,
--CO.sub.2R.sup.2, NR.sup.2--C(O)--R.sup.2;
[0018] each n is independently 1 or 2;
[0019] G, when present, is selected from H, R.sup.7 or
C.sub.1-C.sub.4 alkyl, or, when G is C.sub.1-C.sub.4 alkyl, G and
R' are bound to one another either directly or through a
C.sub.1-C.sub.3 linker to form a heterocyclic ring; or
[0020] when G is not present (i.e., when x in (G).sub.x is 0), then
the nitrogen to which G is attached is bound directly to the
R.sup.7 group on --OR.sup.7;
[0021] D and D' are independently selected from Q; C.sub.1-C.sub.6
alkyl, which is optionally substituted with one or more groups
selected from C.sub.3-C.sub.6 cycloalkyl, --OR.sup.2, --R.sup.3,
--O-Q or Q; C.sub.2-C.sub.4 alkenyl, which is optionally
substituted with one or more groups selected from C.sub.3-C.sub.6
cycloalkyl, --OR.sup.2, --R.sup.3, --O-Q or Q; C.sub.3-C.sub.6
cycloalkyl, which is optionally substituted with or fused to Q; or
C.sub.5-C.sub.6 cycloalkenyl, which is optionally substituted with
or fused to Q;
[0022] each Q is independently selected from a 3-7 membered
saturated, partially saturated or unsaturated carbocyclic ring
system; or a 5-7 membered saturated, partially saturated or
unsaturated heterocyclic ring containing one or more heteroatoms
selected from O, N, S, S(O), or N(R.sup.2); wherein Q is optionally
substituted with one or more groups selected from oxo, --OR.sup.2,
--R.sup.2, --N(R.sup.2).sub.2, --N(R.sup.2)--C(O)--R.sup.2,
--R.sup.2--OH, --CN, --CO.sub.2R.sup.2, --C(O)--N(R.sup.2).sub.2,
halo or --CF.sub.3;
[0023] E is selected from Ht; O-Ht; Ht-Ht; --O--R.sup.3;
--N(R.sup.2)(R.sup.3); C.sub.1-C.sub.6 alkyl, which is optionally
substituted with one or more groups selected from R.sup.4 or Ht;
C.sub.2-C.sub.6 alkenyl, which is optionally substituted with one
or more groups selected from R.sup.4 or Hit; C.sub.3-C.sub.6
saturated carbocycle, which is optionally substituted with one or
more groups selected from R.sup.4 or Ht; or C.sub.5-C.sub.6
unsaturated carbocycle, which is optionally substituted with one or
more groups selected from R.sup.4 or Ht;
[0024] each R.sup.4 is independently selected from --OR.sup.2,
--SR.sup.2, --C(O)--NHR.sup.2, --S(O).sub.2--NHR.sup.2, halo,
--NR.sup.2--C(O)--R.sup.2, --N(R.sup.2).sub.2 or --CN;
[0025] each R.sup.7 is independently selected from
##STR00002##
[0026] wherein each M is independently selected from H, Li, Na, K,
Mg, Ca, Ba, --N(R.sup.2).sub.4, C.sub.1-C.sub.12-alkyl,
C.sub.2-C.sub.12-alkenyl, or --R.sup.6; wherein 1 to 4 --CH.sub.2
radicals of the alkyl or alkenyl group, other than the --CH.sub.2
that is bound to Z, is optionally replaced by a heteroatom group
selected from O, S, S(O), S(O.sub.2), or N(R.sup.2); and wherein
any hydrogen in said alkyl, alkenyl or R.sup.6 is optionally
replaced with a substituent selected from oxo, --OR.sup.2,
--R.sup.2, N(R.sup.2).sub.2, N(R.sup.2).sub.3, R.sup.2OH, --CN,
--CO.sub.2R.sup.2, --C(O)--N(R.sup.2).sub.2,
S(O).sub.2--N(R.sup.2).sub.2, N(R.sup.2)--C(O)--R.sub.2,
C(O)R.sup.2, --S(O).sub.n--R.sup.2, OCF.sub.3,
--S(O).sub.n--R.sup.6, N(R.sup.2)--S(O).sub.2 (R.sup.2), halo,
--CF.sub.3, or --NO.sub.2;
[0027] M' is H, C.sub.1-C.sub.12-alkyl, C.sub.2-C.sub.12-alkenyl,
or --R.sup.6; wherein 1 to 4 --CH.sub.2 radicals of the alkyl or
alkenyl group is optionally replaced by a heteroatom group selected
from O, S, S(O), S(O.sub.2), or N(R.sup.2); and wherein any
hydrogen in said alkyl, alkenyl or R.sup.6 is optionally replaced
with a substituent selected from oxo, --OR.sup.2, --R.sup.2,
--N(R.sup.2).sub.2, N(R.sup.2).sub.3, --R.sup.2OH, --CN,
--CO.sub.2R.sup.2, --C(O)--N(R.sup.2).sub.2,
--S(O).sub.2--N(R.sup.2).sub.2, --N(R.sup.2)--C(O)--R.sup.2,
--C(O)R.sup.2, --S(O).sub.n--R.sup.2, --OCF.sub.3,
--S(O).sub.n--R.sup.6, --N(R.sup.2)--S(O).sub.2 (R.sup.2), halo,
--CF.sub.3, or --NO.sub.2;
[0028] Z is CH.sub.2, O, S, N(R.sup.2).sub.2, or, when M is absent,
H;
[0029] Y is P or S;
[0030] X is O or S; and
[0031] R.sup.9 is C(R.sup.2).sub.2, O or N(R.sup.2); and wherein
when Y is S, Z is not S; and
[0032] R.sup.6 is a 5-6 membered saturated, partially saturated or
unsaturated carbocyclic or heterocyclic ring system, or an 8-10
membered saturated, partially saturated or unsaturated bicyclic
ring system; wherein any of said heterocyclic ring systems contains
one or more heteroatoms selected from O, N, S, S(O).sub.n or
N(R.sup.2); and wherein any of said ring systems optionally
contains 1 to 4 substituents independently selected from OH,
C.sub.1-C.sub.4 alkyl, O--C.sub.1-C.sub.4 alkyl or
OC(O)C.sub.1-C.sub.4 alkyl.
[0033] It is a also an object of this invention to provide
pharmaceutical compositions comprising the sulfonamide prodrugs of
formula I and methods for their use as prodrugs of HIV aspartyl
protease inhibitors.
DETAILED DESCRIPTION OF THE INVENTION
[0034] In order that the invention herein described may be more
fully understood, the following detailed description is set forth.
In the description, the following abbreviations are used:
TABLE-US-00001 Designation Reagent or Fragment Ac acetyl Me methyl
Et ethyl Bzl benzyl Trityl triphenylmethyl Asn D- or L-asparagine
Ile D- or L-isoleucine Phe D- or L-phenylalanine Val D- or L-valine
Boc tert-butoxycarbonyl Cbz benzyloxycarbonyl (carbobenzyloxy) Fmoc
9-fluorenylmethoxycarbonyl DCC dicyclohexylcarbodiimide DIC
diisopropylcarbodiimide EDC 1-(3-dimethylaminopropyl)-3-
ethylcarbodiimide hydrochloride HOBt 1-hydroxybenzotriazole HOSu
1-hydroxysuccinimide TFA trifluoroacetic acid DIEA
diisopropylethylamine DBU 1,8-diazabicyclo (5.4.0) undec-7-ene
EtOAc ethyl acetate
[0035] The following terms are employed herein:
[0036] Unless expressly stated to the contrary, the terms
"--SO.sub.2--" and "--S(O).sub.2-" as used herein refer to a
sulfone or sulfone derivative (i.e., both appended groups linked to
the S), and not a sulfinate ester.
[0037] For the compounds of formula I, and intermediates thereof,
the stereochemistry of OR.sup.7 is defined relative to D on the
adjacent carbon atom, when the molecule is drawn in an extended
zig-zag representation (such as that drawn for compounds of formula
XI, XV, XXII, XXIII and XXXI). If both OR.sup.7 and D reside on the
same side of the plane defined by the extended backbone of the
compound, the stereochemistry of OR.sup.7 will be referred to as
"syn". If OR.sup.7 and D reside on opposite sides of that plane,
the stereochemistry of OR.sup.7 will be referred to as "anti".
[0038] The term "aryl", alone or in combination with any other
term, refers to a carbocyclic aromatic radical containing the
specified number of carbon atoms.
[0039] The term "heterocyclic" refers to a stable 5-7 membered
monocycle or 8-11 membered bicyclic heterocycle which is either
saturated or unsaturated, and which may be optionally benzofused if
monocyclic. Each heterocycle consists of carbon atoms and from one
to four heteroatoms selected from the group consisting of nitrogen,
oxygen and sulfur. As used herein, the terms "nitrogen and sulfur
heteroatoms" include any oxidized form of nitrogen and sulfur, and
the quaternized form of any basic nitrogen. The heterocyclic ring
may be attached by any heteroatom of the cycle which results in the
creation of a stable structure. Preferred heterocycles defined
above include, for example, benzimidazolyl, imidazolyl,
imidazolinoyl, imidazolidinyl, quinolyl, isoquinolyl, indolyl,
pyridyl, pyrrolyl, pyrrolinyl, pyrazolyl, pyrazinyl, quinoxolyl,
piperidinyl, morpholinyl, thiamorpholinyl, furyl, thienyl,
triazolyl, thiazolyl, .beta.-carbolinyl, tetrazolyl, thiazolidinyl,
benzofuranoyl, thiamorpholinyl sulfone, benzoxazolyl,
oxopiperidinyl, oxopyrroldinyl, oxoazepinyl, azepinyl, isoxazolyl,
tetrahydropyranyl, tetrahydrofuranyl, thiadiazoyl, benzodioxolyl,
thiophenyl, tetrahydrothiophenyl and sulfolanyl.
[0040] The terms "HIV protease" and "HIV aspartyl protease" are
used interchangeably and refer to the aspartyl protease encoded by
the human immunodeficiency virus type 1 or 2. In a preferred
embodiment of this invention, these terms refer to the human
immunodeficiency virus type 1 aspartyl protease.
[0041] The term "pharmaceutically effective amount" refers to an
amount effective in treating HIV infection in a patient. The term
"prophylactically effective amount" refers to an amount effective
in preventing HIV infection in a patient. As used herein, the term
"patient" refers to a mammal, including a human.
[0042] The term "pharmaceutically acceptable carrier or adjuvant"
refers to a non-toxic carrier or adjuvant that may be administered
to a patient, together with a compound of this invention, and which
does not destroy the pharmacological activity thereof.
[0043] Pharmaceutically acceptable salts of the compounds of this
invention include those derived from pharmaceutically acceptable
inorganic and organic acids and bases. Examples of suitable acids
include hydrochloric, hydrobromic, sulfuric, nitric, perchloric,
fumaric, maleic, phosphoric, glycollic, lactic, salicylic,
succinic, toluene-p-sulfonic, tartaric, acetic, citric,
methanesulfonic, formic, benzoic, malonic, naphthalene-2-sulfonic
and benzenesulfonic acids. Other acids, such as oxalic, while not
in themselves pharmaceutically acceptable, may be employed in the
preparation of salts useful as intermediates in obtaining the
compounds of the invention and their pharmaceutically acceptable
acid addition salts.
[0044] Salts derived from appropriate bases include alkali metal
(e.g., sodium), alkaline earth metal (e.g., magnesium), ammonium
and N--(C.sub.1-4 alkyl).sup.4+ salts.
[0045] The term "thiocarbamates" refers to compounds containing the
functional group N--SO.sub.2--O.
[0046] The compounds of this invention contain one or more
asymmetric carbon atoms and thus occur as racemates and racemic
mixtures, single enantiomers, diastereomeric mixtures and
individual diastereomers. All such isomeric forms of these
compounds are expressly included in the present invention. Each
stereogenic carbon may be of the R or S configuration. The
explicitly shown hydroxyl is also preferred to be syn to D, in the
extended zigzag conformation between the nitrogens shown in
compounds of formula I.
[0047] Combinations of substituents and variables envisioned by
this invention are only those that result in the formation of
stable compounds. The term "stable", as used herein, refers to
compounds which possess stability sufficient to allow manufacture
and administration to a mammal by methods known in the art.
Typically, such compounds are stable at a temperature of 40.degree.
C. or less, in the absence of moisture or other chemically reactive
conditions, for at least a week.
[0048] The compounds of the present invention may be used in the
form of salts derived from inorganic or organic acids. Included
among such acid salts, for example, are the following: acetate,
adipate, alginate, aspartate, benzoate, benzenesulfonate,
bisulfate, butyrate, citrate, camphorate, camphorsulfonate,
cyclopentanepropionate, digluconate, dodecylsulfate,
ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate,
hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide,
hydroiodide, 2-hydroxy-yethanesulfonate, lactate, maleate,
methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate,
pamoate, pectinate, persulfate, 3-phenylpropionate, picrate,
pivalate, propionate, succinate, tartrate, thiocyanate, tosylate
and undecanoate.
[0049] This invention also envisions the quaternization of any
basic nitrogen-containing groups of the compounds disclosed herein.
The basic nitrogen can be quaternized with any agents known to
those of ordinary skill in the art including, for example, lower
alkyl halides, such as methyl, ethyl, propyl and butyl chloride,
bromides and iodides; dialkyl sulfates including dimethyl, diethyl,
dibutyl and diamyl sulfates; long chain halides such as decyl,
lauryl, myristyl and stearyl chlorides, bromides and iodides; and
aralkyl halides including benzyl and phenethyl bromides. Water or
oil-soluble or dispersible products may be obtained by such
quaternization.
[0050] The novel sulfonamides of this invention are those of
formula I:
##STR00003##
wherein:
[0051] A is selected from H; Ht; --R.sup.1--Ht;
--R.sup.1--C.sub.1-C.sub.6 alkyl, which is optionally substituted
with one or more groups independently selected from hydroxy,
C.sub.1-C.sub.4 alkoxy, Ht, --O-Ht,
--NR.sup.2--CO--N(R.sup.2).sub.2 or --CO--N(R.sup.2).sub.2;
--R.sup.1--C.sub.2-C.sub.6 alkenyl, which is optionally substituted
with one or more groups independently selected from hydroxy,
C.sub.1-C.sub.4 alkoxy, Ht, --O-Ht,
--NR.sup.2--CO--N(R.sup.2).sub.2 or --CO--N(R.sup.2).sub.2; or
R.sup.7;
[0052] each R.sup.1 is independently selected from --C(O)--,
--S(O).sub.2--, --C(O)--C(O)--, --O--C(O)--, --O--S(O).sub.2,
--NR.sup.2--S(O).sub.2--, --NR.sup.2--C(O)-- or
--NR.sup.2--C(O)--C(O)--;
[0053] each Ht is independently selected from C.sub.3-C.sub.7
cycloalkyl; C.sub.5-C.sub.7 cycloalkenyl; C.sub.6-C.sub.10 aryl; or
a 5-7 membered saturated or unsaturated heterocycle, containing one
or more heteroatoms selected from N, N(R.sup.2), O, S and
S(O).sub.n; wherein said aryl or said heterocycle is optionally
fused to Q; and wherein any member of said Ht is optionally
substituted with one or more substituents independently selected
from oxo, --OR.sup.2, SR.sup.2, --R.sup.2, --N(R.sup.2)(R.sup.2),
--R.sup.2--OH, --CN, --CO.sub.2R.sup.2, --C(O)--N(R.sup.2).sub.2,
--S(O).sub.2--N(R.sup.2).sub.2, --N(R.sup.2)--C(O)--R.sup.2,
--C(O)--R.sup.2, --S(O).sub.n--R.sup.2, --OCF.sub.3,
--S(O).sub.n-Q, methylenedioxy, --N(R.sup.2)--S(O).sub.2 (R.sup.2),
halo, --CF.sub.3, --NO.sub.2, Q, --OQ, --OR.sup.7, --SR.sup.7,
--R.sup.7, --N(R.sup.2)(R.sup.7) or --N(R.sup.7).sub.2;
[0054] each R.sup.2 is independently selected from H, or
C.sub.1-C.sub.4 alkyl optionally substituted with Q;
[0055] B, when present, is
--N(R.sup.2)--C(R.sup.3).sub.2--C(O)--;
[0056] each x is independently 0 or 1;
[0057] each R.sup.3 is independently selected from H, Ht,
C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.3-C.sub.6
cycloalkyl or C.sub.5-C.sub.6 cycloalkenyl; wherein any member of
said R.sup.3, except H, is optionally substituted with one or more
substituents selected from --OR.sup.2, --C(O)--NH--R.sup.2, --S(O),
--N(R.sup.2)(R.sup.2), Ht, --CN, --SR.sup.2, --CO.sub.2R.sup.2,
NR.sup.2--C(O)--R.sup.2;
[0058] each n is independently 1 or 2;
[0059] G, when present, is selected from H, R.sup.7 or
C.sub.1-C.sub.4 alkyl, or, when G is C.sub.1-C.sub.4 alkyl, G and
R.sup.7 are bound to one another either directly or through a
C.sub.1-C.sub.3 linker to form a heterocyclic ring; or
[0060] when G is not present (i.e., when x in (G).sub.x is 0), then
the nitrogen to which G is attached is bound directly to the
R.sup.7 group in --OR.sup.7 with the concomitant displacement of
one -ZM group from R.sup.7;
[0061] D and D' are independently selected from Q; C.sub.1-C.sub.6
alkyl, which is optionally substituted with one or more groups
selected from C.sub.3-C.sub.6 cycloalkyl, --OR.sup.2, --R.sup.3,
--O-Q or Q; C.sub.2-C.sub.4 alkenyl, which is optionally
substituted with one or more groups selected from C.sub.3-C.sub.6
cycloalkyl, --OR.sup.2, --R.sup.3, --O-Q or Q; C.sub.3-C.sub.6
cycloalkyl, which is optionally substituted with or fused to Q; or
C.sub.5-C.sub.6 cycloalkenyl, which is optionally substituted with
or fused to Q;
[0062] each Q is independently selected from a 3-7 membered
saturated, partially saturated or unsaturated carbocyclic ring
system; or a 5-7 membered saturated, partially saturated or
unsaturated heterocyclic ring containing one or more heteroatoms
selected from O, N, S, S(O).sub.n or N(R.sup.2); wherein Q is
optionally substituted with one or more groups selected from oxo,
--OR.sup.2, --R.sup.2, --N(R.sup.2).sub.2,
--N(R.sup.2)--C(O)--R.sup.2, --R.sup.2--OH, --CN,
--CO.sub.2R.sup.2, --C(O)--N(R.sup.2).sub.2, halo or
--CF.sub.3;
[0063] E is selected from Ht; O-Ht; Ht-Ht; --O--R.sup.3;
--N(R.sup.2)(R.sup.3); C.sub.1-C.sub.6 alkyl, which is optionally
substituted with one or more groups selected from R.sup.4 or Ht;
C.sub.2-C.sub.6 alkenyl, which is optionally substituted with one
or more groups selected from R.sup.4 or Ht; C.sub.3-C.sub.6
saturated carbocycle, which is optionally substituted with one or
more groups selected from R.sup.4 or Ht; or C.sub.5-C.sub.6
unsaturated carbocycle, which is optionally substituted with one or
more groups selected from R.sup.4 or Ht;
[0064] each R.sup.4 is independently selected from --OR.sup.2,
--SR.sup.2, --C(O)--NHR.sup.2, --S(O).sub.2--NHR.sup.2, halo,
--NR.sup.2--C(O)--R.sup.2, --N(R.sup.2).sub.2 or --CN;
[0065] each R.sup.7 is independently selected from
##STR00004##
[0066] wherein each M is independently selected from H, Li, Na, K,
Mg, Ca, Ba, --N(R.sup.2).sub.4, C.sub.1-C.sub.12-alkyl,
C.sub.2-C.sub.12-alkenyl, or --R.sup.6; wherein 1 to 4 --CH.sub.2
radicals of the alkyl or alkenyl group, other than the --CH.sub.2
that is bound to Z, is optionally replaced by a heteroatom group
selected from O, S, S(O), S(O.sub.2), or N(R.sup.2); and wherein
any hydrogen in said alkyl, alkenyl or R.sup.6 is optionally
replaced with a substituent selected from oxo, --OR.sup.2,
--R.sup.2, N(R.sup.2).sub.2, N(R.sup.2).sub.3, R.sup.2OH, --CN,
--CO.sub.2R.sup.2, --C(O)--N(R.sup.2).sub.2,
S(O).sub.2--N(R.sup.2).sub.2, N(R.sup.2)--C(O)--R.sub.2,
C(O)R.sup.2, --S(O).sub.n--R.sup.2, OCF.sub.3,
--S(O).sub.n--R.sup.6, N(R.sup.2)--S(O).sub.2 (R.sup.2), halo,
--CF.sub.3, or --NO.sub.2;
[0067] M' is H, C.sub.1-C.sub.12-alkyl, C.sub.2-C.sub.12-alkenyl,
or --R.sup.6; wherein 1 to 4 --CH.sub.2 radicals of the alkyl or
alkenyl group is optionally replaced by a heteroatom group selected
from O, S, S(O), S(O.sub.2), or N(R.sup.2); and wherein any
hydrogen in said alkyl, alkenyl or R.sup.6 is optionally replaced
with a substituent selected from oxo, --OR.sup.2, --R.sup.2,
--N(R.sup.2).sub.2, N(R.sup.2).sub.3, --R.sup.2OH, --CN,
--CO.sub.2R.sup.2, --C(O)--N(R.sup.2).sub.2,
--S(O).sub.2--N(R.sup.2).sub.2, --N(R.sup.2)--C(O)--R.sup.2,
--C(O)R.sup.2, --S(O).sub.n--R.sup.2, --OCF.sub.3,
--S(O).sub.n--R.sup.6, --N(R.sup.2)--S(O).sub.2 (R.sup.2), halo,
--CF.sub.3, or --NO.sub.2;
[0068] Z is CH.sub.2, O, S, N(R.sup.2).sub.2, or, when M is not
present, H.
[0069] Y is P or S;
[0070] X is O or S; and
[0071] R.sup.9 is C(R.sup.2).sub.2, O or N(R.sup.2); and wherein
when Y is S, Z is not S; and
[0072] R.sup.6 is a 5-6 membered saturated, partially saturated or
unsaturated carbocyclic or heterocyclic ring system, or an 8-10
membered saturated, partially saturated or unsaturated bicyclic
ring system; wherein any of said heterocyclic ring systems contains
one or more heteroatoms selected from O, N, S, S(O).sub.n or
N(R.sup.2); and wherein any of said ring systems optionally
contains 1 to 4 substituents independently selected from OH,
C.sub.1-C.sub.4 alkyl, O--C.sub.1-C.sub.4 alkyl or
O--C(O)--C.sub.1-C.sub.4 alkyl.
[0073] Preferably, at least one R.sup.7 is selected from:
##STR00005## ##STR00006##
[0074] It will be understood by those of skill in the art that
component M or M' in the formulae set forth herein will have either
a covalent, a covalent/zwitterionic, or an ionic association with
either Z or R.sup.9 depending upon the actual choice for M or M'.
When M or M' is hydrogen, alkyl, alkenyl, or R.sup.6, M or M' is
covalently bound to R.sup.9 or Z. If M is a mono- or bivalent metal
or other charged species (i.e., NH.sub.4.sup.+), there is an ionic
interaction between M and Z and the resulting compound is a
salt.
[0075] When x is 0 in (M).sub.x, Z may be a charged species. When
that occurs, the other M may be oppositely charged to produce a 0
net charge on the molecule. Alternatively, the counter ion may
located elsewhere in the molecule.
[0076] Except where expressly provided to the contrary, as used
herein, the definitions of variables A, R.sup.1-R.sup.4,
R.sup.6-R.sup.9, Ht, B, x, n, D, D', M, Q, X, Y, Z and E are to be
taken as they are defined above for the compounds of formula I.
[0077] According to a preferred embodiment, the compounds of this
invention are those represented by formulas XXII, XXIII or
XXXI:
##STR00007##
wherein A, R.sup.3, R.sup.7, Ht, D, D', x, E are as defined above
for compounds of formula I. For ease of reference, the two R.sup.3
moieties present in formula XXXI have been labeled R.sup.3 and
R.sup.3'.
[0078] For compounds of formula XXII, more preferred compounds are
those wherein:
[0079] A is selected from 3-tetrahydrofuryl-O--C(O)--,
3-(1,5-dioxane)-O--C(O)--, or
3-hydroxy-hexahydrofura[2,3-b]-furanyl-O--C(O)--;
[0080] D' is C.sub.1-C.sub.4 alkyl which is optionally substituted
with one or more groups selected from the group consisting of
C.sub.3-C.sub.6 cycloalkyl, --OR.sup.2, --R.sup.3, --O-Q and Q;
[0081] E is C.sub.6-C.sub.10 aryl optionally substituted with one
or more substituents selected from oxo, --OR.sup.2, SR.sup.2,
--R.sup.2, --N(R.sup.2).sub.2, --R.sup.2--OH, --CN,
--CO.sub.2R.sup.2, --C(O)--N(R.sup.2).sub.2,
--S(O).sub.2--N(R.sup.2).sub.2, --N(R.sup.2)--C(O)--R.sup.2,
--C(O)--R.sup.2, --S(O), --R.sup.2, --OCF.sub.3, --S(O).sub.n-Q,
methylenedioxy, --N(R.sup.2)--S(O).sub.2 (R.sup.2), halo,
--CF.sub.3, --NO.sub.2, Q, --OQ, --OR.sup.7, --SR.sup.7, --R.sup.7,
--N(R.sup.2)(R.sup.7) or --N(R.sup.7).sub.2; or a 5-membered
heterocyclic ring containing one S and optionally containing N as
an additional heteroatom, wherein said heterocyclic ring is
optionally substituted with one to two groups independently
selected from --CH.sub.3, R.sup.4, or Ht.
[0082] Ht, insofar as it is defined as part of R.sup.3, is defined
as above except for the exclusion of heterocycles; and
[0083] all other variables are as defined for formula I.
[0084] Even more preferred are compounds of formula XXII, wherein A
is 3-tetrahydrofuryl-O--C(O)--; G is hydrogen; D' is isobutyl; E is
phenyl substituted with N(R.sup.7).sub.2; each M is independently
selected from H, Li, Na, K, Mg, Ca, Ba, C.sub.1-C.sub.4 alkyl or
--N(R.sup.2).sub.4; and each M' is H or C.sub.1-C.sub.4 alkyl.
[0085] Another preferred embodiment for the formula XXII compounds
are those wherein:
[0086] E is a 5-membered heterocyclic ring containing one S and
optionally containing N as an additional heteroatom, wherein said
heterocyclic ring is optionally substituted with one to two groups
independently selected from --CH.sub.3, R.sup.4, or Ht; and
[0087] all other variables are as defined for formula I.
[0088] Even more preferred are any of the formula XXII compounds
set forth above, wherein R.sup.7 in --OR.sup.7 is --PO(OM).sub.2 or
C(O)CH.sub.2OCH.sub.2CH.sub.2OCH.sub.2CH.sub.2OCH.sub.3 and both
R.sup.7 in --N(R.sup.7).sub.2 are H, wherein M is H, Li, Na, K or
C.sub.1-C.sub.4 alkyl; or wherein R.sup.7 in --OR.sup.7 is
C(O)CH.sub.2OCH.sub.2CH.sub.2OCH.sub.3, one R.sup.7 in
--N(R.sup.7).sub.2 is C(O)CH.sub.2OCH.sub.2CH.sub.2OCH.sub.3 and
the other is H.
[0089] The most preferred compound of formula XXII has the
structure:
##STR00008##
[0090] For compounds of formula XXIII, most preferred compounds are
those wherein:
[0091] R.sup.3 is C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl,
C.sub.5-C.sub.6 cycloalkyl, C.sub.5-C.sub.6 cycloalkenyl or a 5-6
membered saturated or unsaturated heterocycle, wherein any member
of said R.sup.3 may be optionally substituted with one or more
substituents selected from the group consisting of --OR.sup.2,
--C(O)--NH--R.sup.2, --S(O).sub.nN(R.sup.2)(R.sup.2), Ht, --CN,
--SR.sup.2, --C(O).sub.2R.sup.2 and NR.sup.2--C(O)--R.sup.2;
and
[0092] D' is C.sub.1-C.sub.3 alkyl or C.sub.3 alkenyl, wherein said
alkyl or alkenyl may optionally be substituted with one or more
groups selected from the group consisting of C.sub.3-C.sub.6
cycloalkyl, --OR.sup.2, --O-Q and Q (with all other variables being
defined as above for compounds of formula I).
[0093] Even more preferred are compounds of formula XXIII described
above, wherein R.sup.7 is --PO(OM).sub.2 or --C(O)-M'.
[0094] For compounds of formula XXXI, most preferred compounds are
those wherein A is R.sup.1--Ht, each R.sup.3 is independently
C.sub.1-C.sub.6 alkyl which may be optionally substituted with a
substituent selected from the group consisting of --OR.sup.2,
--C(O)--NH--R.sup.2, --S(O).sub.nN(R.sup.2)(R.sup.2), Ht, --CN,
--SR.sup.2, --CO.sub.2R.sup.2 or --NR.sup.2--C(O)--R.sup.2; and D'
is C.sub.1-C.sub.4 alkyl, which may be optionally substituted with
a group selected from the group consisting of C.sub.3-C.sub.6
cycloalkyl, --OR.sup.2, --O-Q; and E is Ht, Ht-Ht and
--NR.sup.2R.sup.3.
[0095] Even more preferred are those compounds of formula XXXI
described above wherein R.sup.7 is --PO(OM).sub.2 or --C(O)-M'.
TABLE-US-00002 TABLE I ##STR00009## CMPD R.sup.7 W 198 ##STR00010##
--NO.sub.2 199 ##STR00011## --NH.sub.2 200 ##STR00012## --NH.sub.2
201 ##STR00013## --NH.sub.2 202 ##STR00014## --NH.sub.2 203
##STR00015## --NH.sub.2 204 ##STR00016## --NH.sub.2 205
##STR00017## --NH.sub.2 206 ##STR00018## --NH.sub.2 207
##STR00019## --NH.sub.2 208 ##STR00020## --NO.sub.2 209
##STR00021## --NO.sub.2 210 ##STR00022## --NH.sub.2 211
##STR00023## --NH.sub.2 212 ##STR00024## --NH.sub.2 213
##STR00025## --NH.sub.2 214 ##STR00026## --NH.sub.2 215
##STR00027## --NH.sub.2 216 ##STR00028## --NH.sub.2 217
##STR00029## --NH.sub.2 219 H ##STR00030## 220 H ##STR00031## 221 H
##STR00032## 222 H ##STR00033## 223 H ##STR00034## 224 H
##STR00035## 225 ##STR00036## ##STR00037## 226 ##STR00038##
--NO.sub.2 227 ##STR00039## --NO.sub.2 228 ##STR00040## --NH.sub.2
229 ##STR00041## --NH.sub.2 230 H ##STR00042## 231 ##STR00043##
##STR00044## 237 ##STR00045## --NO.sub.2 238 ##STR00046##
--NO.sub.2 239 --SO.sub.3H --NO.sub.2 240 --SO.sub.3H --NH.sub.2
241 ##STR00047## --NO.sub.2 242 ##STR00048## --NH.sub.2 245
##STR00049## --NH.sub.2 246 ##STR00050## --NH.sub.2 247
##STR00051## --NH.sub.2 248 ##STR00052## --NH.sub.2 249
##STR00053## --NH.sub.2 250 ##STR00054## --NH.sub.2 251
##STR00055## --NH.sub.2 252 ##STR00056## --NH.sub.2 253
##STR00057## --NH.sub.2 254 ##STR00058## --NH.sub.2 255 H --NH--CHO
256 H ##STR00059## 257 H ##STR00060## 258 H ##STR00061## 259 H
##STR00062## 260 H ##STR00063## 261 ##STR00064## ##STR00065## 262
##STR00066## ##STR00067## 263 ##STR00068## ##STR00069## 264
PO.sub.3K.sub.2 --NH.sub.2 265 PO.sub.3Ca --NH.sub.2 266 PO.sub.3Mg
--NH.sub.2 267 ##STR00070## --NH.sub.2 308 ##STR00071## --NH.sub.2
402 H ##STR00072## 403 H ##STR00073## 404 H ##STR00074## 405 H
##STR00075## 406 H ##STR00076## 407 H ##STR00077## 408 ##STR00078##
--NH.sub.2
TABLE-US-00003 TABLE II ##STR00079## COMPOUND A R.sup.7 232
##STR00080## ##STR00081## 233 H ##STR00082## 234 ##STR00083## H 235
##STR00084## ##STR00085## 236 ##STR00086## ##STR00087##
TABLE-US-00004 TABLE III ##STR00088## COMPOUND R.sup.7 W 243
##STR00089## --NO.sub.2 244 ##STR00090## --NH.sub.2 400
##STR00091## --NO.sub.2 401 ##STR00092## --NH.sub.2
[0096] According to another embodiment, the invention provides
compounds of the following formulae:
##STR00093## ##STR00094##
wherein, in compound 1005, when R.sup.7 is PO.sub.3M, (G).sub.x is
not H; and wherein R.sup.10 is selected from isopropoyl or
cyclopentyl; R.sup.11 is selected from NHR.sup.7 or OR; and x,
R.sup.7 and G are as defined above.
[0097] The prodrugs of the present invention may be synthesized
using conventional synthetic techniques. U.S. Pat. No. 5,585,397
discloses the synthesis of compounds of formula:
##STR00095##
wherein A, B, n, D, D', and E are as defined above. Prodrugs of
formula (I) of the present invention can be readily synthesized
from the '397 compounds using conventional techniques. One of skill
in the art would be well aware of conventional synthetic reagents
to convert the --OH group of the '397 compounds to a desired
--OR.sup.7 functionality of the present invention, wherein R.sup.7
is as defined above. The relative ease with which the compounds of
this invention can be synthesized represents an enormous advantage
in the large scale production of these compounds.
[0098] For example, VX-478, a compound disclosed in the '397
patent, can be readily converted to the corresponding bis-phosphate
ester derivative, as shown below:
##STR00096##
[0099] Alternatively, if the monophosphate ester of VX-478 is
desired, then the synthetic scheme can be readily adapted by
beginning with the 4-nitrophenyl derivative of VX-478, as shown
below:
##STR00097##
[0100] Examples of specific compounds in addition to VX-478 which
may be converted to the prodrugs of this invention by similar
techniques (and the syntheses of those intermediates to the
compounds of the present invention) are disclosed in WO 94/05639
and WO 96/33184, the disclosures of which are herein incorporated
by reference.
[0101] Pharmaceutically acceptable salts of the compounds of the
present invention may be readily prepared using known techniques.
For example, the disodium salt of the mono-phosphate ester shown
above can be prepared as shown below:
##STR00098##
[0102] The compounds of this invention may be modified by appending
appropriate functionalities to enhance selective biological
properties. Such modifications are known in the art and include
those which increase biological penetration into a given biological
system (e.g., blood, lymphatic system, central nervous system),
increase oral availability, increase solubility to allow
administration by injection, alter metabolism and alter rate of
excretion.
[0103] Without being bound by theory, we believe that two different
mechanisms are involved in converting the prodrugs of this
invention into the active drug, depending upon the structure of the
prodrug. The first mechanism involves the enzymatic or chemical
transformation of the prodrug species into the active form. The
second mechanism involves the enzymatic or chemical cleavage of a
functionality on the prodrug to produce the active compound.
[0104] The chemical or enzymatic transformation can involve to
transfer of a functional group (i.e., R.sup.7) from one heteroatom
within the molecule to another heteroatom. This transfer is
demonstrated in the chemical reactions shown below:
##STR00099##
[0105] The cleavage mechanism is demonstrated by the reaction below
where a phosphate ester-containing prodrug is converted into the
active form of the drug by removal of the phosphate group.
##STR00100##
[0106] These protease inhibitors and their utility as inhibitors of
aspartyl proteases are described in U.S. Pat. No. 5,585,397, the
disclosure of which is incorporated herein by reference.
[0107] The prodrugs of the present invention are characterized by
unexpectedly high aqueous solubility. This solubility facilitates
administration of higher doses of the prodrug, resulting in a
greater drug load per unit dosage. The prodrugs of the present
invention are also characterized by facile hydrolytic cleavage to
release the active aspartyl protease inhibitor in vivo. The high
aqueous solubility and the facile in vivo metabolism result in a
greater bioavailability of the drug. As a result, the pill burden
on a patient is significantly reduced.
[0108] The prodrugs of this invention may be employed in a
conventional manner for the treatment of viruses, such as HIV and
HTLV, which depend on aspartyl proteases for obligatory events in
their life cycle. Such methods of treatment, their dosage levels
and requirements may be selected by those of ordinary skill in the
art from available methods and techniques. For example, a prodrug
of this invention may be combined with a pharmaceutically
acceptable adjuvant for administration to a virally-infected
patient in a pharmaceutically acceptable manner and in an amount
effective to lessen the severity of the viral infection.
[0109] Alternatively, the prodrugs of this invention may be used in
vaccines and methods for protecting individuals against viral
infection over an extended period of time. The prodrugs may be
employed in such vaccines either alone or together with other
compounds of this invention in a manner consistent with the
conventional utilization of protease inhibitors in vaccines. For
example, a prodrug of this invention may be combined with
pharmaceutically acceptable adjuvants conventionally employed in
vaccines and administered in prophylactically effective amounts to
protect individuals over an extended period time against HIV
infection. As such, the novel protease inhibitors of this invention
can be administered as agents for treating or preventing HIV
infection in a mammal.
[0110] The prodrugs of this invention may be administered to a
healthy or HIV-infected patient either as a single agent or in
combination with other anti-viral agents which interfere with the
replication cycle of HIV. By administering the compounds of this
invention with other anti-viral agents which target different
events in the viral life cycle, the therapeutic effect of these
compounds is potentiated. For instance, the co-administered
anti-viral agent can be one which targets early events in the life
cycle of the virus, such as cell entry, reverse transcription and
viral DNA integration into cellular DNA. Anti-HIV agents targeting
such early life cycle events include, didanosine (ddI), alcitabine
(ddC), d4T, zidovudine (AZT), polysulfated polysaccharides, sT4
(soluble CD4), ganiclovir, dideoxycytidine, trisodium
phosphonoformate, eflornithine, ribavirin, acyclovir, alpha
interferon and trimenotrexate. Additionally, non-nucleoside
inhibitors of reverse transcriptase, such as TIBO or nevirapine,
may be used to potentiate the effect of the compounds of this
invention, as may viral uncoating inhibitors, inhibitors of
trans-activating proteins such as tat or rev, or inhibitors of the
viral integrase.
[0111] Combination therapies according to this invention exert a
synergistic effect in inhibiting HIV replication because each
component agent of the combination acts on a different site of HIV
replication. The use of such combinations also advantageously
reduces the dosage of a given conventional anti-retroviral agent
which would be required for a desired therapeutic or prophylactic
effect as compared to when that agent is administered as a
monotherapy. These combinations may reduce or eliminate the side
effects of conventional single anti-retroviral agent therapies
while not interfering with the anti-retroviral activity of those
agents. These combinations reduce potential of resistance to single
agent therapies, while minimizing any associated toxicity. These
combinations may also increase the efficacy of the conventional
agent without increasing the associated toxicity. In particular, we
have discovered that these prodrugs act synergistically in
preventing the replication of HIV in human T cells. Preferred
combination therapies include the administration of a prodrug of
this invention with AZT, ddI, ddC or d4T.
[0112] Alternatively, the prodrugs of this invention may also be
co-administered with other HIV protease inhibitors such as Ro
31-8959 (Roche), L-735,524 (Merck), XM 323 (Du-Pont Merck) and
A-80,987 (Abbott) to increase the effect of therapy or prophylaxis
against various viral mutants or members of other HIV quasi
species.
[0113] We prefer administering the prodrugs of this invention as
single agents or in combination with retroviral reverse
transcriptase inhibitors, such as derivatives of AZT, or other HIV
aspartyl protease inhibitors. We believe that the co-administration
of the compounds of this invention with retroviral reverse
transcriptase inhibitors or HIV aspartyl protease inhibitors may
exert a substantial synergistic effect, thereby preventing,
substantially reducing, or completely eliminating viral infectivity
and its associated symptoms.
[0114] The prodrugs of this invention can also be administered in
combination with immunomodulators (e.g., bropirimine, anti-human
alpha interferon antibody, IL-2, GM-CSF, methionine enkephalin,
interferon alpha, diethyldithiocarbamate, tumor necrosis factor,
naltrexone and rEPO); and antibiotics (e.g., pentamidine
isethiorate) to prevent or combat infection and disease associated
with HIV infections, such as AIDS and ARC.
[0115] When the prodrugs of this invention are administered in
combination therapies with other agents, they may be administered
sequentially or concurrently to the patient. Alternatively,
pharmaceutical or prophylactic compositions according to this
invention may be comprised of a combination of a prodrug of this
invention and another therapeutic or prophylactic agent.
[0116] Although this invention focuses on the use of the prodrugs
disclosed herein for preventing and treating HIV infection, the
compounds of this invention can also be used as inhibitory agents
for other viruses which depend on similar aspartyl proteases for
obligatory events in their life cycle. These viruses include, as
well as other AIDS-like diseases caused by retroviruses, such as
simian immunodeficiency viruses, but are not limited to, HTLV-I and
HTLV-II. In addition, the compounds of this invention may also be
used to inhibit other aspartyl proteases, and in particular, other
human aspartyl proteases, including renin and aspartyl proteases
that process endothelin precursors.
[0117] Pharmaceutical compositions of this invention comprise any
of the compounds of the present invention, and pharmaceutically
acceptable salts thereof, with any pharmaceutically acceptable
carrier, adjuvant or vehicle. Pharmaceutically acceptable carriers,
adjuvants and vehicles that may be used in the pharmaceutical
compositions of this invention include, but are not limited to, ion
exchangers, alumina, aluminum stearate, lecithin, serum proteins,
such as human serum albumin, buffer substances such as phosphates,
glycine, sorbic acid, potassium sorbate, partial glyceride mixtures
of saturated vegetable fatty acids, water, salts or electrolytes,
such as protamine sulfate, disodium hydrogen phosphate, potassium
hydrogen phosphate, sodium chloride, zinc salts, colloidal silica,
magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based
substances, polyethylene glycol, sodium carboxymethylcellulose,
polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers,
polyethylene glycol and wool fat.
[0118] The pharmaceutical compositions of this invention may be
administered orally, parenterally, by inhalation spray, topically,
rectally, nasally, buccally, vaginally or via an implanted
reservoir. We prefer oral administration or administration by
injection. The pharmaceutical compositions of this invention may
contain any conventional non-toxic pharmaceutically-acceptable
carriers, adjuvants or vehicles. The term parenteral as used herein
includes subcutaneous, intracutaneous, intravenous, intramuscular,
intra-articular, intrasynovial, intrasternal, intrathecal,
intralesional and intracranial injection or infusion
techniques.
[0119] The pharmaceutical compositions may be in the form of a
sterile injectable preparation, for example, as a sterile
injectable aqueous or oleaginous suspension. This suspension may be
formulated according to techniques known in the art using suitable
dispersing or wetting agents (such as, for example, Tween 80) and
suspending agents. The sterile injectable preparation may also be a
sterile injectable solution or suspension in a non-toxic
parenterally-acceptable diluent or solvent, for example, as a
solution in 1,3-butanediol. Among the acceptable vehicles and
solvents that may be employed are mannitol, water, Ringer's
solution and isotonic sodium chloride solution. In addition,
sterile, fixed oils are conventionally employed as a solvent or
suspending medium. For this purpose, any bland fixed oil may be
employed including synthetic mono- or diglycerides. Fatty acids,
such as oleic acid and its glyceride derivatives are useful in the
preparation of injectables, as are natural
pharmaceutically-acceptable oils, such as olive oil or castor oil,
especially in their polyoxyethylated versions. These oil solutions
or suspensions may also contain a long-chain alcohol diluent or
dispersant such as Ph. Helv or a similar alcohol.
[0120] The pharmaceutical compositions of this invention may be
orally administered in any orally acceptable dosage form including,
but not limited to, capsules, tablets, and aqueous suspensions and
solutions. In the case of tablets for oral use, carriers which are
commonly used include lactose and corn starch. Lubricating agents,
such as magnesium stearate, are also typically added. For oral
administration in a capsule form, useful diluents include lactose
and dried corn starch. When aqueous suspensions are administered
orally, the active ingredient is combined with emulsifying and
suspending agents. If desired, certain sweetening and/or flavoring
and/or coloring agents may be added.
[0121] The pharmaceutical compositions of this invention may also
be administered in the form of suppositories for rectal
administration. These compositions can be prepared by mixing a
compound of this invention with a suitable non-irritating excipient
which is solid at room temperature but liquid at the rectal
temperature and therefore will melt in the rectum to release the
active components. Such materials include, but are not limited to,
cocoa butter, beeswax and polyethylene glycols.
[0122] Topical administration of the pharmaceutical compositions of
this invention is especially useful when the desired treatment
involves areas or organs readily accessible by topical application.
For application topically to the skin, the pharmaceutical
composition should be formulated with a suitable ointment
containing the active components suspended or dissolved in a
carrier. Carriers for topical administration of the compounds of
this invention include, but are not limited to, mineral oil, liquid
petroleum, white petroleum, propylene glycol, polyoxyethylene
polyoxypropylene compound, emulsifying wax and water.
Alternatively, the pharmaceutical composition can be formulated
with a suitable lotion or cream containing the active compound
suspended or dissolved in a carrier. Suitable carriers include, but
are not limited to, mineral oil, sorbitan monostearate, polysorbate
60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl
alcohol and water. The pharmaceutical compositions of this
invention may also be topically applied to the lower intestinal
tract by rectal suppository formulation or in a suitable enema
formulation. Topically-transdermal patches are also included in
this invention.
[0123] The pharmaceutical compositions of this invention may be
administered by nasal aerosol or inhalation. Such compositions are
prepared according to techniques well-known in the art of
pharmaceutical formulation and may be prepared as solutions in
saline, employing benzyl alcohol or other suitable preservatives,
absorption promoters to enhance bioavailability, fluorocarbons,
and/or other solubilizing or dispersing agents known in the
art.
[0124] Dosage levels of between about 0.01 and about 100 mg/kg body
weight per day, preferably between about 0.5 and about 50 mg/kg
body weight per day of the active ingredient compound are useful in
the prevention and treatment of viral infection, including HIV
infection. Typically, the pharmaceutical compositions of this
invention will be administered from about 1 to about 5 times per
day or alternatively, as a continuous infusion. Such administration
can be used as a chronic or acute therapy. The amount of active
ingredient that may be combined with the carrier materials to
produce a single dosage form will vary depending upon the host
treated and the particular mode of administration. A typical
preparation will contain from about 5% to about 95% active compound
(w/w). Preferably, such preparations contain from about 20% to
about 80% active compound.
[0125] Upon improvement of a patient's condition, a maintenance
dose of a compound, composition or combination of this invention
may be administered, if necessary. Subsequently, the dosage or
frequency of administration, or both, may be reduced, as a function
of the symptoms, to a level at which the improved condition is
retained when the symptoms have been alleviated to the desired
level, treatment should cease. Patients may, however, require
intermittent treatment on a long-term basis upon any recurrence of
disease symptoms.
[0126] As the skilled artisan will appreciate, lower or higher
doses than those recited above may be required. Specific dosage and
treatment regimens for any particular patient will depend upon a
variety of factors, including the activity of the specific compound
employed, the age, body weight, general health status, sex, diet,
time of administration, rate of excretion, drug combination, the
severity and course of the infection, the patient's disposition to
the infection and the judgment of the treating physician.
[0127] In order that this invention be more fully understood, the
following examples are set forth. These examples are for the
purpose of illustration only and are not to be construed as
limiting the scope of the invention in any way.
Example 1
General Conditions
[0128] (A) Analytical HPLC 0-100% B/30 min, 1.5 mL/min, A=0.1% TFA
in water, B=0.1% TFA in acetonitrile. Detection at 254 and 220 nm,
C18 reverse phase Vydac, t0=2.4 min.
[0129] (B) 1/3 v/v EtOAc/hexane
[0130] (C) 1/2 v/v EtOAc/hexane
[0131] (D) Analytical HPLC 0-100% B/10 min, 1.5 mL/min, A=0.1% TFA
in water, B=0.1% TFA in acetonitrile. Detection at 254 and 220 nm,
C18 reverse phase Vydac, t.sub.0=2.4 min.
##STR00101##
[0132] A mixture of 2.0 g (3.7 mMol) of 197 and 3.0 g (16 mMol) of
di-p-nitrophenyl carbonate in 10 ml of dimethylformamide was
treated at 25.degree. with 4 ml (4 mMol) of P4-phosphazene base
(Fluka, 1M in hexane). The mixture was stirred for 6 h at
25.degree. until all of the starting alcohol was consumed. The
reaction mixture was partitioned between ethyl acetate and 1N
hydrochloric acid. The organic layer was washed with 1N sodium
hydroxide and brine, dried over magnesium sulfate and concentrated
in vacuo. Titration with dichloromethane gave the desired mixed
carbonate (1.2 g crop 1 and 0.6 g crop 2) as a fine powder.
Combined yield: 69%. Rf=0.13 (1/3 EtOAc/hexane, conditions B),
Rf=0.40 (1/2 EtOAc/hexane, conditions C), tHPLC=23.83 min (A), MS
(ES+) 701 (M+1).
[0133] 1H-NMR (CDCl3): 0.82 (6H, dd), 1.9 (2H, m), 2.15 (1H, m),
2.8 (1H, m), 3.0 (4H, m), 3.5 (2H, m), 3.6 (1H, m), 3.8 (4H, m),
4.3 (1H, bs), 4.8 (1H, m), 5.17 (2H, m), 7.7 (7H, m), 7.95 (2H, d),
8.35 (4H, m).
[0134] 13C (CDCl3): 155.2 152.2, 149.9, 145.6, 135.9, +129.0,
+128.8, +128.5, +127.2, +125.4, +124.4, +121.8, +78.1, +75.8,
-73.1, -66.9, -56.5, +52.7, -48.2, -35.9, -35.9, 32.6, -+26.4,
+19.9, +19.8.
Example 2
##STR00102##
[0136] To 0.20 g (0.286 mM) of 198 dissolved in 3 ml of THF was
added 0.11 g (1.14 mM) of 1-Methyl-piperidine and the mixture was
stirred overnight at room temperature ("rt"). All the solvents were
then evaporated and the solid residue partitioned between EtOAc and
water. The volatiles were removed and, where appropriate, the
residue was treated with 1:1 TFA/DCM over 30 min at rt to remove
the Boc protecting group. The product was dissolved in 0.25 ml TFA
and 1.5 ml THF. Hydrogenolysis for 10 hours in presence of 30 mg of
10% Pd/C gave the desired compound. The final purification was on
preparative reversed phase C18 using conditions Example 1, except
that the flow rate was 18 ml/min.
[0137] C,H,N: calc: 49.27, 5.57, 8.25, found 49.15, 5.76, 8.29.
C.sub.33H.sub.45N.sub.5O.sub.7S.sub.1.1.9CF.sub.3COOH
[0138] LC/MS (ES+) 632 (M+1) 1 peak at 4.71 min
[0139] Analytical HPLC(A) t=N/A min
[0140] 1H, 0.71 (3H, d), 0.74 (3H, d), 1.80 (2H, m), 2.03 (1H, m),
2.63 (2H, m), 2.74 (1H, m), 2.82 (3H, s), 2.92 (2H, m), 3.20 (4H,
m), 3.42 (3H, m), 3.62 (2H, m), 3.75 (1H, m), 4.05 (3H, m), 4.97
(2H, m), 6.2 (1H, bs), 6.60 (2H, m), 7.22 (5H, m), 7.40 (3H,
m),
[0141] 13C (DMSO): 156.4, 154.0, 153.8, 138.8, 129.6, 129.5, 128.3,
126.5, 123.7, 112.7, 74.8, 72.9, 66.7, 58.2, 54.0, 53.1, 49.3,
42.3, 40.8, 36.0, 33.3, 25.8, 20.4, 20.3
Example 3
##STR00103##
[0143] The synthesis of compound 200 from compound 198 was carried
as described in Example 1, except that N,N-dimethyl-aminoethanol
was used in place of di-p-nitrophenyl carbonate.
[0144] 1HNMR (acetone-d6): 0.82 (6H, dd), 1.83 (2H, m), 2.07 (1H,
m), 2.64 (2H, m), 2.82 (6H, s), 2.90 (2H, m), 3.19 (1H, m), 3.38
(4H, m), 3.63 (2H, m), 3.76 (1H, m), 4.17 (2YH, m), 4.40 (1H, m),
4.56 (1H, m), 4.96 (1H, m), 5.06 (1H, m), 6.06 (1H, d), 6.68 (2H,
d), 7.23 (5H, m), 7.47 (2H, d).
[0145] 13CNMR (acetone d6): 20.2, 20.3, 27.5, 33.4, 35.6, 43.8,
50.1, 54.2, 56.4, 58.5, 63.1, 67.4, 73.6, 76.2, 79.9, 114.2, 118.3,
127.4, 129.2, 130.1, 130.3, 139.3, 153.4, 157.0.
[0146] LC/MS: 1 peak, 621 (MH+).
Example 4
##STR00104##
[0148] The synthesis of compound 201 from compound 198 was carried
as described in Example 1, except that N-acetyl-ethylenediamine was
used in place of di-p-nitrophenyl carbonate.
[0149] C,H,N: calc: 49.66, 5.64, 8.83, found 49.76, 5.98, 8.93.
C.sub.30H.sub.43N.sub.5O.sub.8S.sub.1.1.4CF.sub.3COOH.
[0150] LC/MS (ES+) 634 (M+1) 1 peak at 5.08 min.
[0151] Analytical HPLC(A) t=15.92 min.
[0152] 1H: d-3 acetonitrile: 0.88 (6H, dd), 1.92 (3H, s), 1.94 (2H,
m), 2.17 (1H, m), 2.72 (2H, m), 2.96 (2H, m), 3.07 (3H, m), 3.29
(1H, m), 3.42 (3H, m), 3.69 (1H, m), 3.77 (1H, m), 3.82 (1H, m),
4.133 (1H, m), 4.40 (1H, bs), 5.05 (2H, m), 5.80 (1H, m), 6.10 (1H,
d), 6.78 (2H, d), 6.83 (1H, bs), 7.28 (5H, m), 7.58 (2H, d).
[0153] 13C (d3-acetonitrile): 157.1, 157.0, 153.2, 139.6, +130.3,
+130.2, +129.2, +127.2, 126.2, +114.2, +76.0, +75.4, -73.6, -67.4,
-58.2, +54.9, -50.2, -41.6, -39.8, -35.9, -33.4, +27.3, +23.1,
+20.4, +20.2.
Example 5
##STR00105##
[0155] The synthesis of compound 202 from compound 198 was carried
as described in Example 1, except that mono N-Boc-piperazine was
used in place of di-p-nitrophenyl carbonate.
[0156] C,H,N: calc: 48.28, 5.68, 8.41, found 48.28, 5.36, 8.28.
C.sub.30H.sub.43N.sub.5O.sub.7S.sub.1.times.2 CF.sub.3COOH
[0157] LC/MS (ES+) 618 (M+1) 1 peak at 4.36 min.
[0158] Analytical HPLC(A) t=14.84 min.
[0159] 1H: d6-DMSO: 0.72 (3H, d), 0.77 (3H, d), 1.78 (2H, m), 2.09
(1H, m), 2.64 (2H, m), 2.73 (1H, m), 2.80 (1H, m), 3.08 (4H, m),
3.32 (2H, m), 3.41 (1H, m), 3.50 (4H, m), 3.54 (1H, m), 3.63 (1H,
m), 3.70 (1H, m), 3.98 (1H, m), 4.89 (1H, m), 4.97 (1H, m), 6.61
(2H, d), 7.23 (5H, m), 7.42 (3H, m), 8.88 (2H, bs).
[0160] 13C: (DMSO): 155.7, 153.6, 153.0, 138.4, +129.1, +129.0,
+128.1, +126.1, 123.2, +112.7, +75.2, +74.4, -72.5, -66.2, -56.9,
+53.1, -48.8, -42.5, -40.8, -35.0, -32.2, +26.2, +20.0, +19.8.
Example 6
##STR00106##
[0162] The synthesis of compound 203 from compound 198 was carried
as described in Example 1, except that mono-N-Boc-ethylenediamine
was used in place of di-p-nitrophenyl carbonate.
[0163] C,H,N: calc: 46.89, 5.29, 8.54, found 46.50, 5.51, 8.54.
C.sub.28H.sub.41N.sub.5O.sub.7S.sub.1.times.2 CF.sub.3COOH.
[0164] LC/MS (ES+) 592 (M+1) 1 peak at 4.32 min.
[0165] Analytical HPLC(A) t=14.69 min.
[0166] 1H:d-6 DMSO: 0.77 (6H, d), 1.82 (2H, m), 2.06 (1H, m), 2.57
(2H, m), 2.82 (4H, m), 2.97 (1H, m), 3.30 (5H, m), 3.55 (1H, m),
3.65 (1H, m), 3.70 (1H, m), 3.95 (1H, m), 4.88 (1H, m), 4.95 (1H,
m), 6.62 (2H, d), 7.20 (6H, m), 7.39 (3H, m), 7.78 (3H, bs).
[0167] 13C (dmso): 155.9, 152.9, 138.5, 129.2, 128.9, 128.1, 126.1,
122.9, 112.7, 74.7, 74.5, 72.6, 66.2, 57.2, 53.2, 49.4, 38.8,
37.94, 35.1, 32.1, 26.3, 20.0, 19.8.
##STR00107##
Example 7
[0168] The synthesis of compound 204 from compound 1.9 was carried
as described in Example 1, except that
mono-1,3-diamino-3-N-Boc-propane was used in place of
di-p-nitrophenyl carbonate.
[0169] C,H,N: calc: 49.07, 5.64, 8.89, found 48.95, 6.00, 8.92.
C.sub.29H.sub.43N.sub.5O.sub.7S.sub.1.times.1.6CF.sub.3COOH
[0170] LC/MS (ES+) 605 (M+1) 1 peak at 4.27 min.
[0171] Analytical HPLC(A) t=14.72 min.
[0172] 1H:d-6 DMSO: 0.78 (6H, dd), 1.64 (2H, m), 1.83 (2H, m), 2.03
(1H, m), 2.57 (1H, m), 2.78 (4H, m), 2.94 (1H, m), 3.03 (2H, m),
3.32 (2H, m), 3.58 (1H, m), 3.63 (1H, m), 3.73 (1H, m), 3.87 (1H,
m), 4.84 (1H, m), 4.92 (1H, m), 6.61 (2H, d), 7.22 (6H.m), 7.36
(1H, d), 7.28 (2H, d), 7.76 (3H, ns).
[0173] 13C (dmso): 155.8, 155.7, 138.5, +129.1, +129.0, +128.0,
+126.1, 122.9, +112.7, +74.6, +74.3, -72.7, -66.2, -57.2, +53.6,
-49.5, -37.4, -36.7, -35.5, -32.1, -27.6, +26.2, +20.0, +19.8.
Example 8
##STR00108##
[0175] The synthesis of compound 205 from compound 198 was carried
as described in Example 1, except that 1,4-diamino-4-N-Boc-butane
was used in place of di-p-nitrophenyl carbonate.
[0176] C,H,N: calc: 48.17, 5.59, 8.26, found 48.02, 5.96, 8.24.
C.sub.30H.sub.45N5O.sub.7S.sub.1.2CF.sub.3COOH
[0177] LC/MS (ES+) 620 (M+1) 1 peak at 4.36 min.
[0178] Analytical HPLC(A) t=14.93 min.
[0179] 1H: d-6 DMSO: 0.77 (6H, dd), 1.43 (4H, m), 1.82 (2H, m),
2.03 (1H, m), 2.77 (4H, m), 2.95 (3H, m), 3.31 (2H, m), 3.56 (1H,
m), 3.63 (1H, m), 3.70 (1H, bq), 3.82 (1H, m), 4.85 (1H, m), 4.92
(1H, m), 6.62 (2H, d), 7.2 (7H, m), 7.38 (2H, d), 7.72 (3H,
bs).
[0180] 13C: 155.7, 152.9, +138.6, +129.1, +129.0, +128.0, +126.1,
+123.0, +112.7, +74.4, +74.3, -72.7, -66.2, -57.2, +53.7, -49.7,
-38.6, -38.5, -35.4, -32.1, -26.3, +26.2, -24.4, +20.1, +19.9.
Example 9
##STR00109##
[0182] The synthesis of compound 206 from compound 198 was carried
as described in Example 1, except that
(3R)-(+)-3-Boc-aminopyrrolidine was used in place of
di-p-nitrophenyl carbonate.
[0183] C,H,N: calc: 48.28, 5.36, 8.28, found 47.89, 5.53, 8.57.
C.sub.30H.sub.43N.sub.5O.sub.7S.sub.1.times.2 TFA
[0184] LC/MS (ES+) 618 (M+1) 1 peak at 4.32 min.
[0185] Analytical HPLC(A) t=14.31 min.
[0186] 1H and 13C NMR: complex and overlapping mixtures of
rotomers.
Example 10
##STR00110##
[0188] The synthesis of compound 207 from compound 198 was carried
as described in Example 1, except that
(3S)-(-)-3-Boc-aminopyrrolidine was used in place of
di-p-nitrophenyl carbonate.
[0189] LC/MS (ES+) 618 (M+1) 1 peak at 4.19 min.
[0190] Analytical HPLC(A) t=14.75 min.
[0191] 1H and 13C NMR: complex and overlapping mixtures of
rotomers.
Example 11
##STR00111##
[0193] The synthesis of compound 308 from compound 198 was carried
as described in Example 1, except that
N-triphenylmethyl-N,N'-dimethylethanediamine was used in place of
di-p-nitrophenyl carbonate.
[0194] 1H-NMR: 0.76 (6H, dd), 1.65 (2H, m), 1.95 (1H, m), 2.07 (1H,
m), 2.7 (2H, m), 2.75 (3H, s), 2.95 (3H, m), 3.45 (2H, m), 3.7 (4H,
m), 4.2 (2H, bm), 5.05 (2H, bd), 6.62 (2H, d), 7.2 (5H, m), 7.5
(2H, d).
[0195] LC/MS: 1 peak, 620 (MH+).
Example 12
General Procedures
Acylation:
##STR00112##
[0197] To 200 mg (0.37 mM) of 197 dissolved in 5 ml
CH.sub.2Cl.sub.2 was added N-CBz-L-Benzyl tyrosine 183 mg (0.41 mM)
followed by 231 mg (1.12 mM) DCC, followed by 29 mg (0.23 mM) DMAP.
The reaction is stirred at rt for 24 hr. The precipitates present
were removed by filtration. The filtrate was then concentrated in
vacuo. The final compound was purified on preparative reversed
phase C.sub.18 using purification by HPLC C.sub.18 Waters Delta
Prep 3000 Column: YMC-Pack ODS AA 12S05-2520WT 250.times.20 mm I.D.
S-5 mm, 120 .ANG., 0-100% B over 1/2 h, flow=18 ml/min, monitored
at 220 nm, B=0.1% trifluoroacetic acid in acetonitrile, A=0.1%
trifluoroacetic acid in water. Analytical Column: YMC-Pack ODS AA1
2S05-2520WT 250.times.4.6 mmI.D. S-5 mm, 120 .ANG., 0-100% B at 1.5
ml/min. over 1/2 h, monitored at 220 nm, B=0.1% trifluoroacetic
acid in acetonitrile, A=0.1% trifluoroacetic acid in water.
[0198] The aqueous phase was lyophilized to give 59 mg, (16.3%)
GW431896X, (U11484-72-10) t.sub.HPLC=11.71 min., MW=966.04,
LC/MS=MH+967.
Reduction of the Nitro Functionality:
##STR00113##
[0200] A slurry of 209 (170 mg) and 10 mg of 10% Pd.C in 95% EtOH
was flushed with hydrogen in a scintillation vial equipped with
septum and a stir bar. Continuous overnight hydrogenolysis under
hydrogen balloon resulted in a complete conversion. The crude
preparation was then filtered off the catalyst, and purified on RP
C18 HPLC (Prep Nova-Pack C186 um, 60 A, gradient 0-100% B over 30
min. The desired product was collected and lyophilized affording a
white fluffy solid (50 mg, 30.8%).
Example 13
##STR00114##
[0202] Compound 211 was obtained following the acylation and
reduction procedures of Example 12.
[0203] ES+ 669.2 (M+1), tHPLC=8.06 min (D), 13C NMR (DMSO) 168.9,
156.9, 155.7, 153.1, 138.1, 130.5, 129.2, 129.1, 128.1, 126.2,
124.7, 122.5, 112.8, 76.2, 74.5, 72.5, 66.1, 58.0, 53.6, 52.6,
49.2, 33.6, 32.1, 26.6, 25.3, 20.0.
[0204] tHPLC=11.71 min (D), ES+ 967 (M+1).
Example 14
##STR00115##
[0206] 212 was obtained following the procedures of Example 12.
[0207] tHPLC=9.45 min (D), ES+ 592.2 (M+1).
[0208] 13C NMR (DMSO) 171.5, 155.8, 148.9, 137.8, 129.5, 129.3,
128.5, 126.7, 115.2, 75.2, 73.8, 73.1, 68.3, 67.0, 58.7, 57.1,
53.3, 49.2, 35.4, 32.4, 26.7, 20.1, 19.8.
[0209] 1H (CDCl3, 399.42 KHz): 8.33 (2H, d, J=8.8), 7.95 (2H, d,
J=8.8), 7.23 (5H, m) 5.22 (m, 2H), 5.08 (m, 1H), 4.08 (m, 1H),
3.80-3.45 (7H, m), 3.41 (3H, s), 2.98 (m, 3H), 2.66 (m, 1H), 2.57
(m, 2H), 2.10 (s, 1H), 1.93 (2H, m), 0.82 (3H, d), 0.78 (3H,
d).
[0210] ES+ 622 (M+1), 644 (M+Na)
[0211] tHPLC=10.29 min (D).
[0212] 13C NMR (CDCl3): 171.3, 155.5, 149.9, 145.6, 136.9, 129.2,
128.6, 128.5, 126.8, 124.4, 76.7, 75.3, 73.2, 72.9, 68.2, 66.9,
58.7, 55.9, 53.1, 48.3, 35.3, 32.7, 26.3, 19.9, 19.8.
Example 15
##STR00116##
[0214] Compound 213 was obtained following the procedure of Example
12. tHPLC=9.21 min (D); ES+ 622 (M+1).
[0215] 13C NMR (CDCl3): 170.54, 156.2, 148.6, 136.8, 129.4, 129.2,
128.6, 126.6, 115.7, 76.7, 74.6, 73.2, 71.8, 70.6, 68.2, 6.6.9,
58.9, 57.3, 53.8, 49.4, 36.2, 33.1, 26.8, 19.8, 19.5.
[0216] Intermediate: t HPLC=10.05 min (D); ES+=652 (M+H) 674
(M+Na).
Example 16
##STR00117##
[0218] 214 was obtained following the procedure of Example 12.
[0219] ES+ 634.4 (M+1); t HPLC=7.17 min (D).
[0220] 13C (DMSO): 169.3, 155.8, 153.1, 138.0, 129.1, 129.0, 128.1,
126.3, 122.6, 112.8, 94.3, 75.6, 74.6, 72.4, 66.1, 57.8, 52.7,
52.0, 49.3, 38.4, 34.7, 32.2, 29.1, 26.6, 21.4, 20.1, 20.0.
Example 17
##STR00118##
[0222] 215 was obtained following the procedure of Example 12.
[0223] t HPLC=9.12 min (D)
[0224] 1H (DMSO) all signals broad: 7.38 (3H, br m), 7.20 (5H, br
m), 6.62 (2H, br m), 5.15 (1H, br m), 4.92 (1H, br m), 4.00 (3H,
m), 3.7-3.0 (16H, m), 2.78 (2H, m), 2.57 (3H, m), 2.04 (m, 1H),
1.78 (m, 2H), 0.77 (6H, m)
[0225] 13C (DMSO) 170.6, 156.3, 153.7, 139.1, 129.8, 128.4, 126.7,
123.7, 113.3, 79.8, 79.2, 77.3, 76.1, 75.4, 75.2, 73.0, 71.9, 52.3,
51.8, 48.2, 46.7, 39.9, 38.7, 25.8, 22.6.
[0226] Intermediate:
[0227] t HPLC=10.18 min (D); ES+ 696.3 (M+1).
Example 18
##STR00119##
[0229] 216 was obtained following the procedure of Example 12.
[0230] 1H-NMR: 0.97 (6H, t), 1.95 (2H, m), 2.20 (1H, m), 2.9 (2H,
m), 2.96 (6H, s), 3.00 (3H, s), 3.38 (1H, m), 3.42 (3H, m), 3.36
(1H, m), 3.6 (2H, m), 3.7 (6H, m), 3.98 (2H, m), 4.2 (2H, dd), 5.1
(1H, bs), 5.4 (1H, m), 6.8 (2H, d), 7.4 (5H, m), 7.6 (2H, d).
[0231] LC-MS: 1 peak, 692 (MH+).
Example 19
##STR00120##
[0233] 217 was obtained following the procedure of Example 12.
[0234] 1H-NMR (CDCl3): 0.78 (6H, dd), 1.9 (2H, m), 2.1 (1H, m), 2.3
(3H, s), 2.9 (8H, m), 2.9 (2H, m), 3.15 (1H, m), 3.35 (1H, m), 3.5
(1H, m), 3.75 (4H, m), 4.06 (2H, s), 4.15 (2H, m), 4.9 (1H, dd),
5.05 (1H, bs), 5.2 (1H, bs), 6.63 (2H, d), 7.2 (5H, m), 7.55 (2H,
d), 8.0 (2H, m).
[0235] ESMSP: 676 (MH+).
Example 20
General Procedure for N-Acylated Compounds
##STR00121##
[0237] A mixture of 0.5 g (1 mMol) of
(3S)-Tetrahydro-3-furfuryl-N-((1S,2R)-1-benzyl-2-hydroxy-3-(N-isobutyl-4--
aminobenzenesulfonamido)propyl) carbamate, 0.4 g (1.5 mMol) of
Boc-(S)-3-pyridyl alanine, 0.29 g (1.5 mMol) EDCI and 0.1 g
4-dimethylamino pyridine in 10 ml of N,N-dimethylformamide was
stirred at 25.degree. for 12 hours. The volatiles were removed in
vacuo and the residue was partitioned between ethyl acetate and 1N
hydrochloric acid. The organic layer was washed with 1N sodium
hydroxide and brine, dried over magnesium sulfate and concentrated
in vacuo. The residue was chromatographed on a 2 inch plug of
silica gel (1:1 ethyl acetate:hexane) to give the desired
N-acylated material. Deprotection by treatment with 50 ml of
trifluoroacetic acid, followed by co-evaporation of residual acid
with methanol gave the desired prodrug as a white foam (0.2 g,
26%).
[0238] H1-NMR (acetonitrile-D3): 0.95 (6H, dd), 2.0 (2H, m), 2.25
(1 h, m), 2.8-3.1 (5H, m), 3.6-4.0 (7H, m), 4.25 (1H, m), 4.75 (1H,
m), 5.18 (1H, m), 5.45 (1H, m), 7.0 (2H, d), 7.4 (5H, m), 7.75 (2H,
d), 8.2 (1H, m), 8.8 (1H, d), 8.85 (1H, d), 9.15 (1H, s).
[0239] LC/MS: 1 peak, 654 (MH+).
Example 21
##STR00122##
[0241] 220 was obtained using the general procedure in Example
20.
[0242] 1H-NMR (acetone-d6/methanol-d4): 0.95 (6H, t), 2.0 (2H, m),
2.2 (1H, m), 2.90 (1H, dd), 2.95 (2H, d), 3.12 (1H, dd), 3.4 (2H,
m), 6 (1H, d), 3.8 (5H, m), 4.4 (2H, bm), 6.82 (2H, d), 7.20 (1H,
s), 7.4 (5H, m), 7.65 (2H, d), 8.0 (1H, s).
[0243] LC/MS: 1 peak, 643 (MH+).
Example 22
##STR00123##
[0245] 221 was obtained using the general procedure in Example
20.
[0246] 1H-NMR (DMSO d-6): 0.76 (6H, t), 1.80 (2H, m), 2.10 (1H, m),
3.7 (4H, m), 3.75 (3H, s), 3.2 (5H, m), 3.58 (2H, s), 3.7 (4H, m),
4.97 (1H, bm), 5.18 (1H, bs), 6.7 (2H, d), 7.22 (5H, m), 7.45 (2H,
d).
[0247] LC/MS: 1 peak, 646 (MH+).
Example 23
##STR00124##
[0249] 222 was obtained using the general procedure in Example
20.
[0250] 1HNMR (acetonitrile d-3): 1.0 (6H, t), 2.0 (2H, m), 2.2 (1H,
m), 3.00 (6H, s), 3.02 (3H, s), 3.1 (4H, m), 3.5 (3H, m), 3.8 (8H,
m), 4.4 (2H, s), 5.15 (1H, bs), 7.4 (5H, m), 7.97 (2H, d), 8.04
(2H, d).
[0251] LC/MS: 1 peak, 692 (MH+).
Example 24
##STR00125##
[0253] 223 was obtained using the general procedure in Example
20.
[0254] t HPLC=9.22 min (D); ES+ 622 (M+1).
[0255] 1H NMR d6-DMSO: 0.76 (6H, dd), 1.0-1.8 (15H, m), 2.03 (1H,
m), 2.58 (2H, m), 2.79 (2H, m), 3.11 (1H, m), 3.28 (3H, s), 3.3-3.5
(12H, m), 3.94 (1H, m), 4.08 (1H, m), 4.94 (1H, m), 5.14 (1H, m),
6.61 (2H, d), 7.22 (5H, m), 7.40 (3H, m).
[0256] 13C (DMSO) 169.7, 165.9, 152.9, 138.4, 129.2, 129.1, 128.1,
126.2, 123.1, 112.8, 74.4, 74.1, 72.5, 71.2, 69.8, 66.1, 58.1,
57.1, 52.9, 47.5, 33.4, 33.2, 26.3, 24.5, 18.9, 18.8.
Example 25
##STR00126##
[0258] 224 was obtained using the general procedure in Example
20.
Example 26
O,N-Diacylated Prodrugs
[0259] The general procedure for N,O-diacylated compounds followed
the protocol outlined in Example 20, above, except that a five fold
excess of reagents was used relative to the starting material.
##STR00127##
[0260] t HPLC 9.26 min (D); ES+ 738 (M+1) 760 (M+Na).
[0261] 13C (DMSO): 170.2, 169.8, 156.4, 143.4, 138.8, 129.5, 128.8,
128.5, 126.8, 119.7, 74.9, 74.2, 73.7, 71.6, 70.7, 70.3, 68.0,
67.2, 59.3, 57.6, 53.8, 49.6, 35.7, 33.8, 27.1, 20.4.
[0262] 1H (DMSO): 10.1 (1H, s), 7.84 (d, 2H, J=8.5), 7.76 (d,
J=8.7, 2H), 7.40 (1H, d, J=9.2), 7.22 (m, 5H), 5.14 (1H, m), 4.95
(1H, m), 4.1 (m, 8H), 3.7-3.3 (m, 13H), 3.28 (s, 3H), 3.26 (s, 3H),
2.86 (m, 2H), 2.73 (m, 1H), 2.59 (m, 1H), 2.04 (m, 1H), 1.83 (m,
2H), 0.78 (m, 6H).
Example 27
##STR00128##
[0264] To a mixture of 197 (2.93 g, 5.47 mmol) and phosphorous acid
(Aldrich, 2.2 equiv., 12.03 mmol, 987 mg) in 20 ml pyridine was
added 1,3-dicyclohexylcarbodiimide (Aldrich, 2.1 equiv., 11.49
mmol, 2.37 g) and the reaction heated to 60.degree. C. under
nitrogen for 3 h. Solvent was removed in vacuo, the residue treated
with 200 ml 0.1N aqueous sodium bicarbonate and stirred 1 h at
ambient temperature. The mixture was filtered, the filtrate
acidified to pH 1.5 by addition of conc. HCl and extracted with
ethyl acetate (3.times.100 ml). The combined organic layers were
dried over magnesium sulfate, filtered and concentrated in vacuo to
give 3.15 g (96%) of desired product 226 which was used directly in
the next reaction. HPLC: Rt=8.91 min (96%), MS (AP+) 600.5
(M+1).
Example 28
##STR00129##
[0266] A suspension of 226 (.about.5.47 mmol) in 18 ml
hexamethyldisilazane was stirred at 120.degree. C. until
homogeneous followed by addition of bis(trimethylsilyl) peroxide
(Gelest, Inc., 2.3 equiv., 12.58 mmol, 2.24 g, 2.71 ml). After 1 h
the mixture was cooled to ambient temperature, solvent removed in
vacuo, the residue stirred with 100 ml methanol, solvent removed in
vacuo, the residue stirred with 100 ml 0.1N aqueous sodium
bicarbonate, acidified to pH 1.5 by addition of conc. HCl,
saturated with brine and extracted with ethyl acetate (3.times.100
ml). The combined organic layers were dried over magnesium sulfate,
filtered and concentrated in vacuo to give 2.98 g (88%) of desired
product 227, which was used directly in the next reaction. HPLC:
Rt=9.28 min (90%), MS (AP+) 616.5 (M+1).
[0267] Alternatively, 227 can be synthesized directly from 197. In
this method, 197 was dissolved in pyridine (300 mL). The resulting
solution was concentrated in vacuo to about 150 ml at 50-55.degree.
C. The solution was then cooled under N.sub.2 to 5.degree. C., and
treated with POCl.sub.3 (6.5 ml, 1.24 equiv.) over 2 minutes. The
cooling bath was removed and the reaction stirred at ambient
temperature for 2.5 hrs. The solution was then cooled to 5.degree.
C. and water (300 ml) was added over 30 minutes.
[0268] The resulting mixture was extracted with
4-methylpentan-2-one (MIBK, 2.times.150 ml). The combined extracts
were washed with 2N HCl (2.times.250 ml). The acid washes were back
extracted with MIBK (60 ml), then the combined MIBK solutions were
treated with 2N HCl (150 ml). The two phase mixture was stirred
rapidly and heated to 50.degree. C. for 2 hours. The reaction
mixture was cooled to 20.degree. C., the phases were separated and
the MIBK solution was washed with brine (150 ml). The product, 227,
was isolated by drying the solution with magnesium sulfate,
filtering of the drying agent and concentrating in vacuo at
40.degree. C. to give the product as a pale yellow foam (31 g, 90%
yield).
Example 29
##STR00130##
[0270] A solution of 227 (2.98 g, 4.84 mmol) in 50 ml ethyl acetate
was treated with 10% palladium on carbon (Aldrich, 300 mg) and put
under 35 psi of hydrogen on a Parr shaker for 15 h. Catalyst was
removed by filtration and solvent removed in vacuo to give 2.66 g
(94%) of desired product 228. HPLC: Rt=7.23 min (92%), MS (ES+)
586.3 (M+1).
Example 30
##STR00131##
[0272] Solid 228 (2.66 g, 4.54 mmol) was treated with ml aqueous
sodium bicarbonate (Baker, 3.0 equiv., 13.63 mmol, 1.14 g) and
loaded onto a resin column (Mitsubishi Kasei Corp., MCI-gel,
CHP-20). Distilled water was run through until the eluent was
neutral followed by product elution with 1% acetonitrile in water.
Pure fractions were pooled and lyophilized to give 918 mg of pure
bis-sodium salt 229.
[0273] Alternatively, 7 g of 228 was dissolved in 100 ml of EtOAc
with warming and the solution was extracted with 100 ml of aqueous
250 mM triethylammonium bicarbonate (TEABC) (2.times.). The aqueous
extracts were combined and diluted to 1500 ml with water. This
solution was applied to a 300 ml DEAE-52 column (Whatman) which was
equilibrated with 50 mM TEABC. The column was washed with 8 L of 50
mM TEABC and the TEA salt was eluted with 2 L of 250 mM TEABC. The
solution was evaporated en vacuo to 100 ml then lyophilized to
yield the TEA salt (1.5 TEA equivalents). The TEA salt was (5.8 g)
was dissolved in 200 ml water, 300 ml of 1 N HCl was added and the
mixture was extracted with EtOAc (3.times.200 ml). The ethyl
acetate solution was dried with MgSO.sub.4 then evaporated en vacuo
to yield 4 g of the free acid. Two grams of the free acid was
dissolved in 50 ml of acetonitrile and a solution of 573 mg
NaHCO.sub.3 in 200 ml water was added. The mixture was lyophilized
yielding 2.1 g of the bis sodium salt (compound 229).
Example 31
##STR00132##
[0275] 0.53 g (3.0 mmol) 2-[2-(2-Methoxyethoxy)ethoxy]acetic acid
was added to a stirred solution of 1.2 g (3.15 mmol) HATU 0.2 g
(1.47 mmol) HOAt 0.4 g (4.0 mmol) NMM in 10 ml anhydrous
N,N-dimethylformamide. The mixture was stirred at room temperature
for 30 minutes, then 0.5 g (1 mmol) of
(3S)-Tetrahydro-3-furfuryl-N-((1S,2R)-1-benzyl-2hydroxy-3-(N-iso-
butyl-4-aminobenzenesulfonamido)-propyl) carbamate was added to the
solution in one portion. The mixture was stirred at 20.degree. C.
for an hour then at 50.degree. C. for an additional 12 hours. It
was then cooled to 20.degree. C., 50 ml of ether was added, and the
solution was washed with water three times. The aqueous phase was
washed with ether, and then the combined organic phases were dried
with anhydrous magnesium sulfate and filtered. The filtrate was
concentrated under reduced pressure and the residue was purified by
silica gel chromatography to obtain the desired Mono-(N)acylated
(102 mg, 15%) and Bis-(O,N) acylated (262 mg, 32%) compounds.
[0276] Mono-(N)-acylated: 1H-NMR (CDCl3): 0.85 (dd, 6H), 1.85 (m,
2H), 2.08 (m, 1H), 2.8-3.1 (m, 7H), 3.33 (s, 3H), 3.55 (m, 3H),
3.70-3.90 (m, 8H), 4.1 (s, 2H), 5.0 (d, 1H), 5.08 (s(br), 1H), 7.2
(m, 5H), 7.70 (d, 2H), 7.80 (d, 2H), 9.09 (s, 1H).
[0277] MS (FAB+): 666 (M+1).
[0278] Bis-(O,N)-acylated: 1H-NMR (CDCl3): 0.77 (m, 6H), 1.81 (m,
1H), 1.95 (m, 1H), 2.05 (m, 1H), 2.6-3.0 (m, 6H), 3.2 (m, 1H),
3.332 (s, 3H), 3.338 (s, 3H), 3.5-3.8 (m, 18H), 4.1 (s, 2H), 4.14
(s, 2H), 4.17 (m, 1H), 5.05 (m, 2H), 5.25 (s(br), 1H), 7.2 (m, 5H),
7.69 (d, 2H), 7.78 (d 2H), 9.06 (s, 1H).
[0279] MS (FAB+): 826 (M+1), 848 (M+Na).
Example 32
##STR00133##
[0281] We dissolved 0.521 g (1 mM) of 1273W94 in 5 ml THF, then
cooled to -78.degree. C. under nitrogen, and added 1.56 ml (2.5 mM)
of a 1.6 M solution of nBuLi in hexane. After 20 min at -78.degree.
C., we added 105 .mu.L (1.1 mM) of ethyl chlorocarbamate and warmed
up the reaction to room temperature, followed by addition of
another 105 .mu.L of ethyl chlorocarbamate.
[0282] After stirring for additional 4 hrs, the reaction was
quenched with water and the organic solvent evaporated. Part of the
crude product was purified on a silica gel (Rf=0.69 (1:2 ethyl
acetate:hexane)), yielding 0.131 g of the product.
[0283] C,H,N: calc: 46.06, 4.97, 5.88, found 45.90, 4.97, 5.88.
C.sub.23H.sub.33N.sub.5O.sub.5S.sub.1. 2.2 TFA
[0284] LC/MS (ES+) 594 (M+1) 1 peak at 6.96 min.
[0285] Analytical HPLC(A) t=24.57 min. 13C (CDCl3): 155.8, 154.4,
149.9, 145.7, 136.8, +129.2, +128.7, +126.8, +124.2, 80.1, +76.9,
-64.3, -56.2, -52.5, -48.7, -36.2, +28.1, +26.4, +20.0, +19.8,
+14.3.
Example 33
##STR00134##
[0287] We dissolved 0.131 g of the above ethyl carbonate in 4 ml
DCM, followed by 4 ml of TFA. Solvents were then removed after 45
min at room temperature, resulting in the title compound.
[0288] 1H (DMSO): 8.37 (2H, d, J=7.2), 8.15 (2H, m), 8.00 (2H, d,
J=7.0), 7.37 (5H, m), 5.04 (1H, d, J=6.9), 4.06 (2H, q, J=7.0),
3.82 ((1H, m), 3.35 (2H, m), 2.95 (4H, m), 1.82 (1H, m), 1.20 (3H,
t, J=7.0), 0.72 (overlapping doublets, 6H, J=6.2).
[0289] LC/MS 1 peak at 4.76 min.
[0290] ES+ 497.3 (M+1).
Example 34
##STR00135##
[0292] C,H,N: calc: 53.26, 6.14, 7.57, found 53.22, 6.14, 7.57.
C.sub.23H.sub.33N.sub.5O.sub.5S.sub.1.times.0.8 TFA
[0293] LC/MS (ES+) 594 (M+1) 1 peak at 6.96 min.
[0294] Analytical HPLC(A) t=24.57 min. 1H (DMSO): 8.34 (2H, d,
J=8.7), 8.02 (2H, d, J=8.0), 7.19 (5H, m), 6.98 (1H, d, J=7.2),
5.00 (1H, m), 3.83 (2H, q), 3.50 (2H, m), 3.06 (m, 2H), 2.96 (2H,
m), 2.43 (1H, m), 1.97 (1H, m), 1.02 (3H, t), 0.84 (3H, d), 0.82
(3H, d).
[0295] 13C (DMSO): 156.2, 150.1, 145.7, 140.0, +129.7, +129.2,
+128.5, +126.3, +125.0, +71.8, -60.0, +56.2, -56.0, -51.8, -36.0,
+26.3, +20.3, +20.1, +14.6.
Example 35
##STR00136##
[0297] Synthesis of 235 was accomplished analogous to that set
forth in Example 1.
[0298] Yield 15.2%; tHPLC=25.2 min (A).
[0299] Rf=0.54 (B); ES+ 687.3 (M+1).
[0300] 1H (CDCl3): 8.34 (overlapping d+d, 4H), 7.97 (d, 2H, J=8.9),
7.35 (7H, m), 5.09 (1H, m), 4.56 (1H, d, J=8.4), 4.20 (1H, m), 3.54
(1H, m), 3.00 (3H, m), 2.82 (1H, m), 1.84 (1H, m), 1.37 (9H, s),
0.84 (3H, d), 0.82 (3H, d).
Example 36
##STR00137##
[0302] We dissolved 150 mg of 235 in 3 ml of anhydrous dioxane,
added 0.35 ml of S(+)-3-OH-THF and 0.14 ml triethyl amine. The
mixture was refluxed gently under nitrogen for 2 days. Conversion
to 236 was quantitative. Solvents were removed and the compound
purified on silica (B).
[0303] tHPLC=22.98 min (A); ES+ 636.2 (M+1).
[0304] 1H NMR (CDCl3): 8.29 (2H, d), 7.91 (2H, d), 7.22 (5H, m),
5.13 (1H, m), 4.96 (1H, m), 4.52 (1H, d), 4.02 (1H, m), 3.84 (2H,
m), 3.44 (1H, m), 3.36 (1H, m), 3.10 (3H, m, overlap), 2.88 (2H,
m), 2.64 (1H, m), 2.14 (1H, m), 2.05 (1H, m), 1.84 (1H, m), 1.27
(9H, s), 0.78 (6H, two overl. d).
Example 37
Carbohydrate-Based Prodrugs
##STR00138##
[0306] A mixture of 0.54 g (1 mMol) of
(3S)-Tetrahydro-3-furfuryl-N-((1S,2R)-1-benzyl-2-hydroxy-3-(N-isobutyl-4--
aminobenzenesulfonamido)propyl) carbamate, 0.46 g (2 mMol) of
5-dimethyl-tert-butyosilyloxypentanoic acid, 0.346 g (1.8 mMol) of
EDCI and 0.556 mL (4 mMol) of triethylamine in 10 ml of dimethyl
formamide was stirred at rt for 24 h. Another 3 mMol each of acid,
EDCI and triethylamine were added and stirring was continued for an
additional 96 h. A third batch of acid and EDCI was added (3 mMol
each) and the mixture was stirred 72 h to complete the
reaction.
[0307] The reaction mixture was then diluted with ethyl acetate and
extracted with 1N hydrochloric acid, saturated sodium bicarbonate
and water. Evaporation of the solvent and purification on silica
gel (30% ethyl acetate-hexane) gave the desired product (500 mg) as
a waxy solid.
[0308] LCMS: 1 peak, 772.5 (M+Na)
[0309] 1H NMR (CDCL3): 0.01 (6H, s), 0.78 (6H, dd), 0.95 (9H, s),
1.4-1.8 (6H, m), 1.9 (2H, m), 2.05 (1H, m), 2.3 (2H, m), 2.65 (1H,
m), 2.95 (2H, m), 3.22 (1H, m), 3.4 (1H, m), 3.6 (2H, m), 3.75 (3H,
m), 4.8 (1H, d), 5.1 (1H, bs), 5.2 (1H, bs), 7.2 (5H, m), 7.95 (2H,
d), 8.36 (2H, d).
[0310] 450 mg of the 238 was dissolved in 30 ml of tetrahydrofuran
and treated with 20 ml of water and 50 ml of acetic acid. The
mixture was stirred at rt for 2 h and evaporated. Titration with
hexane gave the desired alcohol (290 mg) as a white solid.
[0311] A mixture of 0.15 g (0.24 mMol) of the alcohol produced
above from the previous reaction, 0.205 g (0.5 mMol) of
tetraacetylglucosylbromide and 0.191 g (0.7 mMol) of silver
carbonate in 3 ml of dichloromethane was stirred at rt for 6 h. 150
mg of additional glucosyl bromide and 150 mg of silver carbonate
were added and the mixture was stirred at rt overnight. The mixture
was loaded onto a pad of silica gel and eluted with 30%
ethylacetate-hexane to afford the desired protected carbohydrate
pro-drug as a white foam (200 mg).
[0312] LCMS: 1 peak, 966 (M+H).
[0313] 1H-NMR (CDCl3): 0.78 (6H, dd), 1.9 (2H, m), 2.00 (3H, s),
2.02 (3H, s), 2.05 (3H, s), 2.06 (3H, s), 2.1 (2H, m), 2.3 (2H, m),
2.7 (1H, m), 2.94 (3H, bd), 3.35 (2H, m), 3.45 (2H.m), 3.8 (5H, m),
4.1 (3H, m), 4.5 (1H, d), 4.9 (1H, bs), 4.95 (1H, t), 5.08 (4H, m),
2H, d), 8.35 (2H, d).
Example 38
##STR00139##
[0315] 1.5 g (9.4 mmol) SO3.py complex was added to a stirred
solution of 1 g (1.87 mmol) of 197 in 25 mL anhydrous
tetrahydrofurane. The mixture was stirred at 20.degree. C. for 12
hours, then filtered. The filtrate was concentrated at reduced
pressure, and the residue was transferred to a silica gel column
and eluted with EtOAc (neat), followed by EtOAc:EtOH (4:1) to
obtain 471 mg (47%) 239 as a colorless foam.
[0316] 1H-NMR (CDCl3): 0.80 (m, 6H), 1.8-2.1 (m, 3H), 4.15 (s(br),
1H), 4.8 (t, 1H), 5.04 (s (br), 1H).
[0317] MS (ES-): 614 (M-1).
##STR00140##
[0318] 100 mg (0.162 mmol) 239 dissolved in 15 ml anhydrous
tetrahydrofuran and 200 mg Pd/BaSO4 (5%) was added to the solution.
The mixture was stirred under atmospheric pressure of hydrogen for
8 hours, and then the catalyst was filtered. The filtrate was
concentrated under reduced pressure then dried under vacuum
(.about.1 Hg mm, 48 hrs.) to produce 80 mg (81%) 240 as a colorless
foam.
[0319] 1H-NMR (DMSO-d6): 0.85 (dd, 6H), 0.90 (m, 1H), 2.05 (m, 2H),
2.58 (m, 3H), 2.84 (dd, 1H), 3.05 (m, 2H), 3.55-3.80 (m, 6H), 4.20
(t, 1H), 4.42 (m, 1H), 4.93 (s(br), 1H), 6.09 (s, 2H), 6.70 (d,
2H), 6.80 (d, 1H), 7.15-7.40 (m, 4H), 7.51 (d, 2H).
[0320] MS (ES-): 584 (M-1).
Example 39
##STR00141##
[0322] 780 mg (3 mmol) 2-Chloro-1,3,2-dioxaphospholane was added to
a stirred solution of 1.07 g (2 mmol) 197 and 0.7 ml (4 mmol)
N,N-Diisopropylethylamine in 25 ml dichloromethane at 0.degree. C.
The mixture was allowed to warm up to room temperature and it was
stirred for 2 hours. The mixture was then cooled to 0.degree. C.
and 1.5 g (9.3 mmol) bromine was added in 5 ml dichloromethane. The
mixture was stirred for 1 hour at 20.degree. C., followed by
evaporation under reduced pressure. An aqueous solution (50%) of 15
ml trimethylamine was added to the residue, and the mixture was
stirred at 20.degree. C. for 12 hours.
[0323] Solvents were removed under reduced pressure and 50 ml
EtOAc:EtOH (9:1) was added to the residue. The solid was filtered,
washed with EtOAc:EtOH (9:1) then the filtrate was concentrated
under reduced pressure. The residue was chromatographed on a 3 inch
plug of silica gel using ethyl acetate (neat), then methanol
(neat), as eluents to obtain 1.15 g (82%) 241 as an off-white
solid.
[0324] 1H-NMR (CDCl3): 0.60 (dd, 6H), 1.70 (m, 1H), 1.95 (m, 1H),
2.10 (m, 1H), 2.8-3.2 (m, 6H), 3.4 (s (br), 9H), 5.09 (s(br), 1H),
7.25 (m, 5H), 7.83 (d, 2H), 8.28 (d, 2H).
[0325] MS (ES+): 701 (M+1), 184 (phosphatidyl choline+).
Example 40
##STR00142##
[0327] 250 mg Pd/C (10%) was added to a solution of 250 mg (0.35
mmol) 241 in 10 ml methanol, and the mixture was stirred under
atmospheric pressure of hydrogen for 4 hours at 20.degree. C. The
mixture was filtered, and the filtrate was concentrated under
reduced pressure. The residue was then dissolved in 10 ml water and
lyophilized to obtain 174 mg (74%) 242 as white solid.
[0328] 1H-NMR (DMSO-d6): 0.82 (dd, 6H), 1.80-2.00 (m, 2H), 2.10 (m,
1H), 2.80 (m, 3H), 3.00 (m, 2H), 3.2 (s (br), 9H), 4.0-4.3 (m, 4H),
4.91 (s(br), 1H), 6.08 (s(br), 2H), 6.67 (d, 2H), 7.30 (m, 5H),
7.48 (d, 2H), 8.12 (d, 1H).
[0329] MS (ES+): 671 (M+1), 184 (phosphatidyl choline+).
Example 41
##STR00143##
[0331] 0.175 ml (2 mmol) phosphorus trichloride was added to a
stirred solution of 1.07 g (2 mmol) 197 and 0.35 ml (2 mmol)
N,N-Diisopropylethylamine in 25 ml dichloromethane at 20.degree. C.
The mixture was stirred for 4 hours at 20.degree. C., then 1 ml
water was added and stirred for an additional 12 hours at
20.degree. C. 3 g anhydrous magnesium sulfate was added to the
mixture and it was stirred for minutes, then filtered. The filtrate
was concentrated under reduced pressure and purified by silica gel
chromatography using EtOAc:Hexane (4:1), then EtOAc:EtOH (1:1), to
obtain 402 mg (48%) 226 and 427 mg (36%) 243. 226:
[0332] 1H-NMR (DMSO-d6): 0.82 (dd, 6H), 1.84 (m, 1H), 1.98 (m, 1H),
2.10 (m, 1H), 2.68 (dd, 1H), 2.9-3.2 (m, 4H), 3.6-3.8 (m, 3H), 3.94
(t, 1H), 4.30, (s(br), 1H), 4.97 (s(br), 1H), 7.30 (m, 5H), 8.14
(d, 2H), 8.43 (d, 2H).
[0333] MS (ES-): 598 (M-1).
[0334] 243: (1:1 mix of diastereomers):
[0335] 1H-NMR (CDCl3): 0.80 (m, 6H), 1.8-2.1 (m, 4H), 2.8-3.2 (m,
6H), 3.7-3.9 (m, 4H), 4.15 (m, 1H), 4.8-5.15 (m, 2H), 5.57, 5.72
((d, d), 1H), 7.25 (m, 5H), 7.95 (dd, 2H), 8.35 (m, 2H).
[0336] MS (ES-): 580 (M-1), 598 ((M+H2O)-1).
Example 42
##STR00144##
[0338] The reduction was carried out as described in Example 40;
(Yield: 79%).
[0339] 1H-NMR (DMSO-d6): 0.81 (dd, 6H), 1.82 (m, 1H), 1.95 (m, 1H),
2.08 (m, 1H), 2.6-3.15 (m, 6H), 3.6-3.75 (m, 3H), 4.03 (t, 1H),
4.28, (m, 1H), 4.96 (s(br), 1H), 6.07 (s, 2H), 6.65 (d, 2H), 7.25
(m, 5H), 7.42 (d, 2H).
[0340] MS (ES-): 568 (M-1).
Example 43
##STR00145##
[0342] The reduction was carried out as described in Example 40;
(Yield: 98%).
[0343] (1:1 mix of diastereomers):
[0344] 1H-NMR (DMSO-d6): 0.82 (m, 6H), 1.75-2.0 (m, 2H), 2.05 (m,
1H), 2.6-3.2 (m, 6H), 3.55-3.8 (m, 4H), 4.02, 4.22 (m, t, 1H), 4.75
(m, 1H), 4.90, 5.01 ((d, d), 1H), 6.12 (s, 1H), 6.68 (d, 2H), 7.30
(m, 5H), 7.49 (d, 2H).
[0345] MS (ES-): 550 (M-1), 568 ((M+H2O)-1).
Example 44
Pharmacokinetics in Sprague-Dawley Rats Following Single Oral
Dose
[0346] In order to study the pharmacokinetics of the prodrugs of
this invention, we administered single oral doses of a series of
prodrugs of this invention, as well as VX-478, to male and female
Sprague-Dawley rats. Administration of molar equivalents of a
series of prodrugs of this invention in a variety of pharmaceutical
vehicles was tested.
[0347] Separate groups of male and female Sprague-Dawley rats
(3/sex/group) received oral doses of compound 229 by oral gavage,
in different vehicles at the same dose equivalent (40 mg/kg molar
equivalent of VX-478). The different vehicles for compound 229
were: 1) water; 2) 5/4/1; 3) PEG 400; 4) TPGS/PEG 400; and 5) PEG.
The vehicles for VX-478 were: 1) 33% TPGS/PEG400/PEG; and 2) 12.5%
TPGS/PEG 400/PEG.
[0348] Blood samples were collected following administration at
various time intervals and analyzed for the presence of both
compound 229 and its metabolite, VX-478, by HPLC and MS methods.
The results of this study are tabulated below (Table IV).
TABLE-US-00005 TABLE IV Compound 229 229 229 229 VX-478 VX-478
vehicle H.sub.2O H.sub.2O:PG:EtOH PEG 400 TPGS/PEG 33% TPGS/ 12.5%
TPGS/ 5:4:1 400/PG PEG 400/PG PEG 400/PG number of rats 3 3 3 3 6 3
Molar equiv. 40 PO 40 PO 40 PO 40 PO 41 PO 50 PO dose/478 Dose
(mg/Kg) AUC 11.7 .+-. 4.8 10.6 .+-. 7.4 7.4 .+-. 1.8 8.2 .+-. 1.6
29.6 .+-. 5.8 16.2 .+-. 1.8 (ug*hr/ml) Cmax (.mu.M) 7.1 .+-. 1.7
3.3 .+-. 0.6 3.1 .+-. 0.3 3.0 .+-. 0.7 14.0 .+-. 2.2 6.0 .+-. 1.0
half life (hr) 1.7* 3.4* 2.8* 2.8* 2.5 .+-. 0.9 2.2 .+-. 1.0
Relative Avail. of 39.5.dagger. 35.8.dagger. 25.0.dagger.
27.7.dagger. reference reference VX-478 90.2.dagger..dagger.
81.8.dagger..dagger. 57.1.dagger..dagger. 63.3.dagger..dagger. a
dose of 50 mg/kg of compound 229 is equal to 40 mg/Kg of VX-478. no
compound 229 was detected in plasma at 15 min. (first data point).
*Represents the harmonic mean .dagger.Relative availability of
VX-478 when compared to a prototype clinical formulation
.dagger..dagger.Relative availability of VX-478 when compared to a
prototype toxicology formulation
[0349] We performed a similar study on dogs using both a solid
capsule formulation of compound 229 and an ethanolic/methyl
cellulose solution formulation, as compared to a TPGS-containing
solution formulation of VX-478. The results from this study are
presented below in Table V.
TABLE-US-00006 TABLE V Compound 229 229 VX-478 vehicle solid methyl
22% capsule cellulose in 5% TPGS/PEG EtOH/water 400/PG number of
dogs 2 2 >2 Molar equiv. dose/478 17 PO 17 PO 17 PO Dose (mg/Kg)
AUC 16.7 .+-. 2.7 14.2 .+-. 3.2 23.5 .+-. 7.4 (ug*hr/ml) Cmax
(.mu.g/ml) 6.1 .+-. 1.7 6.3 .+-. 0.3 6.8 .+-. 1.1 Tmax (hr) 2.3
.+-. 0.6 0.5 .+-. 0.5 1.0 .+-. 0.8 Relative Avail. 71.1 60.4
reference of VX-478 (%)
[0350] The results demonstrate that oral administration of compound
229 as an aqueous solution resulted in improved bioavailability in
comparison to the other vehicles studied. Also, following
administration of compound 229, none of that compound was detected
in the first time point blood sample (or later samples), suggesting
first pass metabolism to VX-478. Comparison of the aqueous dose of
compound 229 with the two non-aqueous formulations used for VX-478
indicated equivalence in delivery as illustrated by the range found
for the bioavailability.
Example 45
##STR00146##
[0352] We added 0.28 ml (3.0 mmol) POCl.sub.3 to a stirred solution
of 1.07 g (2.0 mmol) of compound 197 in 10 ml anhydrous pyridine at
5.degree. C. The mixture was allowed to warm up to room temperature
and stirred at 20.degree. C. for 3 hours. The mixture was cooled to
0.degree. C., and quenched with 10 ml water. The solvents were
removed under reduced pressure, the residue was dissolved in 100 ml
ethyl acetate and washed with 20 ml 1M sodium bicarbonate solution.
The organic phase was dried with anhydrous magnesium sulfate,
filtered then concentrated. Chromatographic purification (SiO2,
EtOAc) produce 280 mg of compound 400 (Yield=23%).
[0353] 1H-NMR (DMSO-d6): 0.86 (dd, 6H), 2.05 (m, 2H), 2.84 (d, 2H),
2.95 (dd, 1H), 3.06 (m, 1H), 3.25 (dd, 1H), 3.50-3.70 (m, 4H), 4.20
(m, 1H), 4.35 (m, 1H), 7.2-7.4 (m, 5H), 7.9-8.1 (m, 2H), 8.40 (m,
2H).
[0354] MS (ES-): 596 (M-1).
##STR00147##
[0355] Compound 400 was converted to compound 401 using the
standard hydrogenation method described above employing H2/PdC
(10%), atmospheric pressure, 4 hours at room temperature, solvent:
MeOH--H.sub.2O (5:1). Yield of 401=68%.
[0356] 1H-NMR (DMSO-d6): 0.85 (dd, 6H), 2.0 (m, 2H), 2.6-3.1 (m,
4H), 4.15 (m, 1H), 4.40 (m, 1H), 6.1 (s(br), 1H), 6.61 m (2H),
7.2-7.5 (m, 7H).
[0357] MS (ES-): 566 (M-1).
Example 46
##STR00148##
[0359] We added 1.0 g (2.8) mmol Na-t-Boc-nd-Cbz-L-Ornithine was
added to stirred solution of 1.2 g (3.15 mmol) HATU, 0.2 g (1.47
mmol) HOAt, 0.4 g (4.0 mmol) NMM in 10 ml DMF. The mixture was
stirred at room temperature for 2 hrs. then 0.5 g (1.0 mmol) of
compound 218 was added and the solution was stirred at 50.degree.
C. for 12 hours. The mixture was cooled to room temperature, 100 ml
ether was added and extracted with 5.times.50 ml water. The organic
phase was dried with anhydrous magnesium sulfate, filtered and
concentrated under reduced pressure. The residue was purified by
silica gel chromatography (Hexane-EtOAc (1:1) then EtOAc (neat)) to
yield 410 mg (48%) of compound 350.
Compound 350 A
[0360] 1H-NMR (CDCl3): 0.85 (dd, 6H), 1.41 (s, 3H), 1.45 (s, 6H),
1.60 (m, 4H), 1.90 (m, 2H), 2.1 (m, 1H), 2.75-3.25 (m, 6H),
3.60-3.90 (m, 6H), 5.15 (dd, 2H), 7.2-7.4 (m, 10H), 7.68 (dd,
4H).
[0361] MS (ES-): 852 (M-1).
[0362] MS (ES+): 854 (M+1).
[0363] Compound 350 B
[0364] 1H-NMR (CDCl3): 0.81 (dd, 6H), 1.39 (s, 9H), 1.40-2.10 (m,
9H), 2.70-3.20 (m, 8H), 3.60-3.90 (m, 6H), 4.10 (m, 1H), 4.80 (d,
1H), 5.04 (s(br), 2H), 7.1-7.3 (m, 10H), 7.61 (s, 4H).
[0365] MS (ES-): 866 (M-1).
[0366] MS (ES+): 868 (M+1).
[0367] Compound 350 C
[0368] 1H-NMR (CDCl3): 0.86 (dd, 6H), 1.40 (s, 3H), 1.46 (s, 6H),
1.60-2.10 (m, 7H), 2.70-3.15 (m, 6H), 3.60 (d, 1H), 3.70-4.10 (m,
6H), 4.81 (d, 1H), 5.05-5.30 (m, 7H), 7.18-7.4 (m, 17H), 7.55 (d,
2H).
##STR00149##
[0369] Compounds 350A, 350B and 350C were converted to Compounds
402, 403, and 404, respectively, using the standard hydrogenation
method set forth above:
[0370] H2/PdC (10%), atmospheric pressure, 4 hours, room
temperature, solvent: EtOH, Yield: 81%.
[0371] Compound 402
[0372] 1H-NMR (CDCl3): 0.80 (dd, 6H), 1.38 (s, 9H), 1.8 (m, 6H),
2.10 (m, 2H), 2.75-3.30 (m, 8H), 3.50-4.00 (m, 7H), 4.55 (s(br),
1H), 7.2 (m, 5H), 7.60 (d, 2H), 7.81 (d, 2H).
[0373] MS (ES+): 720 (M+1).
[0374] Compound 403
[0375] 1H-NMR (CDCl3): 0.87 (dd, 6H), 1.45 (s, 9H), 1.50-2.00 (m,
8H), 2.08 (m, 1H), 2.75-3.15 (m, 8H), 3.60 (d, 1H), 3.75-3.90 (m,
5H), 4.28 (s(br), 1H), 4.92 (d, 1H), 5.11 (m, 1H), 5.27 (s(br),
1H), 7.28-7.35 (m, 5H), 7.70 (s, 4H).
[0376] MS (ES+): 734 (M+1).
[0377] Compound 404
[0378] 1H-NMR (CDCl3): 0.80 (dd, 6H), 1.32 (s, 9H), 1.50-2.10 (m,
7H), 2.60-3.20 (m, 8H), 3.40-3.80 (m, 5H), 5.0 (s(br), 1H),
7.05-7.2 (m, 5H), 7.50-7.80 (m, 4H).
[0379] MS (ES+): 762 (M+1).
Example 47
##STR00150##
[0381] We added 5 ml TFA to a stirred solution of 260 mg (0.3 mmol)
Compound 350A, 350B, or 350C in 20 ml chloroform. The mixture was
stirred for 5 hours at room temperature, and then the solvents were
removed under reduced pressure. The residue was dissolved in 20 ml
dichloromethane, 2 ml (11 mmol) N,N-diisopropylethylamine and 1 ml
(10 mmol) acetic anhydride was added to the reaction mixture. The
solution was stirred for 1 hour, then the solvents were removed.
The residue was purified by silica gel chromatography (eluant:
EtOAc-EtOH (9:1)) to obtain 170 mg (71%) of compound 351A, 351B or
351C, respectively.
[0382] Compound 351A
[0383] 1H-NMR (CDCl3): 0.85 (dd, 6H), 1.60 (m, 3H), 1.80-2.00 (m,
3H), 2.06 (2, 3H), 2.75 (dd, 1H), 2.80-3.20 (m, 5H), 3.60-3.90 (m,
7H), 4.85 (d, 2H), 5.10 (m, 3H), 6.46 (d, 1H), 7.25 (m, 10H), 7.67
(s, 4H), 9.30 (s, 1H).
[0384] MS (ES+): 796 (M+1), 818 (M+Na).
[0385] Compound 351B
[0386] 1H-NMR (CDCl3): 0.80 (dd, 6H), 1.38 (m, 2H), 1.50 (m, 2H),
1.70 (m, 0.2H), 1.85 (m, 2H), 2.00 (s, 3H), 2.70 (dd, 1H),
2.75-3.20 (m, 7H), 3.55 (d, 1H), 3.75 (m, 6H), 4.45 (q, 1H), 4.83
(d, 1H), 4.95 (t, 1H), 5.03 (s(br), 3H), 6.46 (d, 1H), 7.20 (m,
10H), 7.61 (s, 4H), 9.29 (s, 1H).
[0387] MS (ES+): 810 (M+1), 832 (M+Na).
[0388] Compound 351C
[0389] 1H-NMR (CDCl3): 0.85 (dd, 6H), 1.70-2.00 (m, 6H), 2.07 (s,
3H), 2.70 (dd, 1H), 2.80-3.00 (m, 3H), 3.10 (dd, 1H), 3.60 (d, 1H),
3.65-4.00 (m, 6H), 4.1 (m, 1H), 4.62 (q, 1H), 4.82 (d, 1H),
5.00-5.30 (m, 5H), 7.10-7.40 (m, 15H), 7.55 (d, 2H), 7.65 (m, 3H)
9.18 (s(br), 1H), 9.45 (s(br), 1H), 9.56 (s(br), 1H).
[0390] MS (FAB+): 972 (M+1), 994 (M+Na).
##STR00151##
[0391] The conversion of compounds 351A, 351C, and 351C to 405,
406, and 407, respectively was achieved by standard hydrogenation
using H2/PdC (10%), atmospheric pressure, 4 hours at room
temperature, solvent: EtOH,
[0392] Yield=46%.
[0393] Compound 405
[0394] 1H-NMR (DMSO-d6): 0.85 (dd, 6H), 1.62 (m, 3H), 1.81 (m, 2H),
1.94 (s, 3H), 2.00-2.2 (m, 2H), 2.75-3.00 (m, 5H), 3.10 (m, 2H),
3.50-3.80 (m, 5H), 4.54 (m, 1H), 5.00 (m, 1H), 5.11 (d, 1H),
7.2-7.4 (m, 5H), 7.80-8.00 (m, 5H), 10.72 (s, 1H).
[0395] MS (ES+): 662 (M+1).
[0396] Compound 406
[0397] 1H-NMR (DMSO-d6): 0.80 (dd, 6H), 1.30-1.80 (m, 7H), 1.85 (s,
3H), 1.95-2.10 (m, 2H), 2.70 (m, 4H), 2.99 (m, 2H), 3.30 (m, 5H),
3.40-3.80 (m, 4H), 4.35 (m, 1H), 4.90 (s, 1H), 5.00 (d, 1H),
7.08-7.25 (m, 5H), 7.50 (s(br), 1H), 7.71 (d, 2H), 7.79 (d, 2H),
10.54 (s, 1H).
[0398] MS (ES+): 676 (M+1).
[0399] Compound 407
[0400] 1H-NMR (DMSO-d6): 0.80 (dd, 6H), 1.40-1.60 (m, 4H), 1.75 (m,
2H), 1.86 (s, 3H), 2.00 (m, 2H), 2.75 (dt, 2H), 3.00 (m, 2H), 3.10
(q, 2H), 3.40-3.70 (m, 5H), 4.39 (q, 1H), 4.92 (s (br), 1H), 5.01
(d, 1H), 7.20 (m, 5H), 7.70 (d+m, 3H), 7.81 (d, 2H), 8.30 (d, 1H),
10.60 (s, 1H).
[0401] MS (ES+): 704 (M+1).
Example 48
##STR00152##
[0403] We added 1.0 g (7.5 mmol) methanephosphonyl dichloride to a
stirred solution of 2.14 g (4.00 mmol) of compound 197 in 20 ml
toluene, containing 10% pyridine. The mixture was stirred at
100.degree. C. for 5 hours, then cooled to 40.degree. C., 2 g (18.5
mmol) benzyl alcohol was added to the reaction, and the mixture was
stirred at 20.degree. C. for 12 hours. The solid was filtered,
washed with 2.times.10 ml toluene and the filtrate was concentrated
under reduced pressure. The residue was purified using silica gel
chromatography (eluants: Hexane-EtOAc (1:1), then EtOAc (neat)) to
yield 550 mg (20%) of compound 352.
[0404] 1H-NMR (CDCl3): 0.67 (dd, 6H), 1.53 (d, 3H), 1.70 (m, 1H),
1.90-2.10 (m, 2H), 2.65-3.20 (m, 6H), 3.55 (d, 1H), 3.80 (m, 3H),
4.10 (m, 1H), 4.70 (q, 1H), 4.90-5.20 (m, 4H), 6.37 (d, 1H),
7.2-7.4 (m, 10H), 7.90 (d, 2H), 8.30 (d, 2H).
[0405] MS (ES+): 704 (M+1), 726 (M+Na).
##STR00153##
[0406] Compound 352 was converted to compound 408 using standard
hydrogenation method: H2/PdC (10%), atmospheric pressure, 2 hours,
room temperature, solvent: MeOH; Yield: 78%.
[0407] 1H-NMR (DMSO-d6): 0.84 (dd, 6H), 1:44 (d, 3H), 1.82 (m, 1H),
1.90-2.10 (m, 2H), 2.62 (m, 2H), 2.95 (m, 2H), 3.10 (d, 1H), 3.39
(d, 1H), 3.45-3.80 (m, 4H), 4.14 (t, 1H), 4.53 (m, 1H), 5.00 (s
(br), 1H), 6.68 (d, 2H), 7.2-7.4 (m, 5H), 7.50 (d, 2H).
[0408] MS (ES-): 582 (M-1).
[0409] While we have described a number of embodiments of this
invention, it is apparent that our basic constructions may be
altered to provide other embodiments which utilize the products and
processes of this invention. Therefore, it will be appreciated that
the scope of this invention is to be defined by the appended
claims, rather than by the specific embodiments which have been
presented by way of example.
* * * * *