U.S. patent application number 09/929773 was filed with the patent office on 2002-01-03 for use of mkc-442 in combination with other antiviral agents.
Invention is credited to Barry, David W., Borroto-Esoda, Katyna, Furman, Phillip A., Moxham, Cary P..
Application Number | 20020002166 09/929773 |
Document ID | / |
Family ID | 21919000 |
Filed Date | 2002-01-03 |
United States Patent
Application |
20020002166 |
Kind Code |
A1 |
Furman, Phillip A. ; et
al. |
January 3, 2002 |
Use of MKC-442 in combination with other antiviral agents
Abstract
Use of MKC-442 in combination with other antiviral agents for
the treatment of patients infected with HIV is disclosed.
Inventors: |
Furman, Phillip A.; (Durham,
NC) ; Moxham, Cary P.; (Durham, NC) ; Barry,
David W.; (Chapel Hill, NC) ; Borroto-Esoda,
Katyna; (Raleigh, NC) |
Correspondence
Address: |
KING & SPALDING
191 PEACHTREE STREET, N.E.
ATLANTA
GA
30303-1763
US
|
Family ID: |
21919000 |
Appl. No.: |
09/929773 |
Filed: |
August 14, 2001 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
09929773 |
Aug 14, 2001 |
|
|
|
09056548 |
Apr 7, 1998 |
|
|
|
60041909 |
Apr 7, 1997 |
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Current U.S.
Class: |
514/252.01 ;
514/263.36 |
Current CPC
Class: |
A61P 37/04 20180101;
A61K 45/06 20130101; A61K 31/513 20130101; A61K 31/505 20130101;
A61P 31/18 20180101; A61K 31/513 20130101; A61K 2300/00
20130101 |
Class at
Publication: |
514/252.01 ;
514/261 |
International
Class: |
A61K 031/522; A61K
031/496 |
Claims
We claim:
1. A method for the treatment of a patient infected with HIV,
comprising administering an effective treatment amount of MKC-442
or its pharmaceutically acceptable salt in combination or
alternation with a protease inhibitor, or its pharmaceutically
acceptable salt.
2. A method for the treatment of a patient infected with HIV,
comprising administering an effective treatment amount of MKC-442
or its pharmaceutically acceptable salt in combination or
alternation with a compound selected from the group consisting of:
indinavir, nelfinavir, saquinavir, amprenavir, and DMP-450 or its
pharmaceutically acceptable salt.
3. The method of claim 2, wherein the compound is indinavir.
4. The method of claim 2, wherein the compound is nelfinavir.
5. The method of claim 2, wherein the compound is saquinavir.
6. The method of claim 2, wherein the compound is amprenavir.
7. The method of claim 2, wherein the compound is DMP-450.
8. A method for the treatment of a patient infected with HIV,
comprising administering an effective treatment amount of MKC-442
or its pharmaceutically acceptable salt in combination or
alternation with a compound selected from the group consisting of
abacavir, FTC, or 3TC or its pharmaceutically acceptable salt.
9. The method of claim 8, wherein the compound is abacavir.
10. The method of claim 8, wherein the compound is FTC.
11. The method of claim 8, wherein the compound is 3TC.
12. A method for the treatment of a patient infected with HIV,
comprising administering an effective treatment amount of MKC-442
or its pharmaceutically acceptable salt in combination or
alternation with a compound selected from the group consisting of
DMP-266, delavirdine, and nevirapine or its pharmaceutically
acceptable salt.
13. The method of claim 12, wherein the compound is DMP-266.
14. The method of claim 12, wherein the compound is
delavirdine.
15. The method of claim 12, wherein the compound is nevirapine.
16. A pharmaceutical composition for the treatment of a patient
infected with HIV, comprising an effective treatment amount of
MKC-442 or its pharmaceutically acceptable salt in combination with
a protease inhibitor, or its pharmaceutically acceptable salt.
17. A pharmaceutical composition for the treatment of a patient
infected with HIV, comprising an effective treatment amount of
MKC-442 or its pharmaceutically acceptable salt in combination or
alternation with a compound selected from the group consisting of:
indinavir, nelfinavir, saquinavir, amprenavir, and DMP-450 or its
pharmaceutically acceptable salt.
18. The composition of claim 17, wherein the compound is
indinavir.
19. The composition of claim 17, wherein the compound is
nelfinavir.
20. The composition of claim 17, wherein the compound is
saquinavir.
21. The composition of claim 17, wherein the compound is
amprenavir.
22. The composition of claim 17, wherein the compound is
DMP-450.
23. A pharmaceutical composition for the treatment of a patient
infected with HIV, comprising an effective treatment amount of
MKC-442 or its pharmaceutically acceptable salt in combination or
alternation with a compound selected from the group consisting of
abacavir, FTC, or 3TC or its pharmaceutically acceptable salt.
24. The composition of claim 8, wherein the compound is
abacavir.
25. The composition of claim 8, wherein the compound is FTC.
26. The composition of claim 8, wherein the compound is 3TC.
27. A pharmaceutical composition for the treatment of a patient
infected with HIV, comprising an effective treatment amount of
MKC-442 or its pharmaceutically acceptable salt in combination with
a compound selected from the group consisting of DMP-266,
delavirdine, and nevirapine or its pharmaceutically acceptable
salt.
28. The composition of claim 27, wherein the compound is
DMP-266.
29. The composition of claim 27, wherein the compound is
delavirdine.
30. The composition of claim 27, wherein the compound is
nevirapine.
Description
[0001] This application claims priority to U.S. Ser. No.
60/041,909, filed on Apr. 7, 1997, by Phillip A. Furman, Carey P.
Moxham, David W. Barry and Katyna Dorroto-Esodo.
[0002] In 1983, the etiological cause of AIDS was determined to be
the human immunodeficiency virus (HIV). In 1985, it was reported
that the synthetic nucleoside 3'-azido-3'-deoxythymidine (AZT)
inhibits the replication of human immunodeficiency virus. Since
then, a number of other synthetic nucleosides, including 2',
3'-dideoxyinosine (DDI), 2', 3'-dideoxycytidine (DDC), and 2',
3'-dideoxy-2', 3'-didehydrothymidine (D4T), have been proven to be
effective against HIV. After cellular phosphorylation to the
5'-triphosphate by cellular kinases, these synthetic nucleosides
are incorporated into a growing strand of viral DNA, causing chain
termination due to the absence of the 3'-hydroxyl group. They can
also inhibit the viral enzyme reverse transcriptase.
[0003] The success of various synthetic nucleosides in inhibiting
the replication of HIV in vivo or in vitro has led a number of
researchers to design and test nucleosides that substitute a
heteroatom for the carbon atom at the 3'-position of the
nucleoside. European Patent Application Publication No. 0 337 713
and U.S. Pat. No. 5,041,449, assigned to BioChem Pharma, Inc.,
disclose racemic 2-substituted-4-substituted-1,3-di- oxolanes that
exhibit antiviral activity. U.S. Pat. No. 5,047,407 and European
Patent Application No. 0 382 526, also assigned to BioChem Pharma,
Inc., disclose that a number of racemic 2-substituted-5-substitut-
ed-1,3-oxathiolane nucleosides have antiviral activity, and
specifically report that the racemic mixture of
2-hydroxymethyl-5-(cytosin-1-yl)-1,3-o- xathiolane (referred to
below as BCH-189) has approximately the same activity against HIV
as AZT, with little toxicity. The (-)-enantiomer of the racemate
BCH-189, known as 3TC, which is covered by U.S. Pat. No. 5,539,116
to Liotta et al., is currently sold for the treatment of HIV in
combination with AZT in humans in the U.S.
[0004] It has also been disclosed that
cis-2-hydroxymethyl-5-(5-fluorocyto- sin-1-yl)-1,3-oxathiolane
("FTC") has potent HIV activity. Schinazi, et al., "Selective
Inhibition of Human Immunodeficiency Viruses by Racemates and
Enantiomers of
cis-5-Fluoro-1-[2-(Hydroxymethyl)-1,3-Oxathiolane-5-yl- ]Cytosine"
Antimicrobial Agents and Chemotherapy, Nov. 1992, pp. 2423-2431.
See also U.S. Pat. No. 5,210,085; WO 91/11186, and WO 92/14743.
[0005] Another compound that exhibits efficacy against HIV both in
vitro and in vivo is 6-benzyl-1-(ethoxymethyl)-5-isopropyluracil,
which is also known as MKC-442. 1
[0006] MKC-442 is described, for example, in U.S. Pat. No.
5,461,060.
[0007] MKC-442, although a nucleoside analogue, functions as a
non-nucleoside reverse transcriptase inhibitor. It is considered an
allosteric inhibitor because it appears to exert its activity by
binding to an "allosteric position", i.e., one other than the
binding site, of the enzyme. Preclinical tests suggest that MKC-442
may possess characteristics that address several of the therapeutic
challenges of HIV. When tested in cell culture assay systems
against wild-type (drug-sensitive) and several mutant strains of
HIV known to be resistant to established non-nucleoside reverse
transcriptase inhibitors, MKC-442 retained much of its ability to
inhibit HIV replication. In these studies, MKC-442 displayed
greater potency than nevirapine against wild-type and mutant
strains of HIV. Preclinical studies of MKC-442 in two drug
combinations with AZT or with DDI and in three drug combinations
with AZT and saquinavir have demonstrated synergistic inhibition of
HIV replication.
[0008] Studies in animals suggest a favorable safety and
pharmacokinetic profile for MKC-442. Animal pharmacokinetic
analyses showed good oral bioavailability and excellent penetration
into the central nervous system, a significant site of HIV
replication that is poorly penetrated by many currently marketed
anti-HIV drugs. In rats, for example, the concentration of MKC-442
in the brain was 100% of that seen in the plasma.
[0009] A Phase I study evaluated the pharmacokinetics and tolerance
of single escalating doses of MKC-442 in HIV-infected volunteers.
The compound was generally well tolerated, with only a few
participants experiencing minor adverse effects at the higher dose
levels. In the groups receiving higher doses, concentrations of the
drug in the plasma reached levels much higher than the levels
required to suppress 90% of the virus in culture.
[0010] Preliminary data from a Phase I/II double-blind, placebo
controlled trial designed to evaluate the safety and efficacy of
repeated multiple oral doses of MKC-442 in HIV-infected patients
has now also been evaluated. A total of 49 patients were treated
with MKC-442 for up to two months. Doses ranging from 100 mg to
1000 mg twice a day were given to groups of six to eight patients
at each dosage level. At the highest doses tested (705 mg and 1000
mg twice a day), the viral load was reduced by an average of 96% in
all patients after one week. This reduction was mostly sustained at
two weeks whereafter it was followed by a gradual increase in viral
load from the nadir toward baseline levels. A single point mutation
at position 13 of the reverse transcriptase that may be associated
with resistance was found in the virus obtained from some patients.
In over 308 patient-weeks of drug exposure, MKC-442 was well
tolerated.
[0011] It is known that over a period of time, agents such as
MKC-442 that are active against HIV induce mutations in the virus
which reduce the efficacy of the drug. There is a need to improve
the durability of antiviral efficacy produced by antiretroviral
drugs, including MKC-442, by decreasing the rate at which such
mutations arise. Further, although MKC-442 exhibits a favorable
pharmacokinetic and biodistribution profile, there is always a
desire to improve these parameters. There is also a need to
decrease the metabolism of the drug, which can lead to an increase
in the plasma concentration of or exposure to MKC-442.
[0012] U.S. Pat. No. 5,604,209, issued on Feb. 18, 1997 to Ubasawa
et al., and assigned to Mitsubishi Chemical Corporation, discloses
that certain 6-benzyl-1-ethoxymethyl-5-substituted uracil
derivatives, including MKC-442, and certain 2',
3'-dideoxyribonucleosides, including 2', 3'-dideoxyinosine (DDI),
3'-azido-3'-deoxythymidine (AZT), AZT triphosphate, and 2',
3'-dideoxycytidine (DDC), exhibit a synergistic effect against
HIV.
[0013] Japanese Patent Application No. 9-18384 filed on Jan. 31,
1997, by Mitsubishi Chemical Corporation, discloses a method for
the treatment of HIV that includes the administration of a
6-benzyl-1-ethoxymethyl-5-subst- ituted uracil derivative,
including MKC-442, in combination with two or more nucleoside-type
reverse transcriptase inhibitors or their esters, and in
particular, those selected from the group consisting of AZT, 2',
3'-dideoxy-3'-thiacytidine (3TC), PMEA
(9-(2-phosphonylmethoxyethyl) adenine (Gilead); PMPA:
(R)-9-(2-phosphonyl-methoxypropyl)adenine); 1592U89 succinate
((1S,4R)-4-[2-amino-6-cyclopropyl-amino)-9H-purin-9-yl]-
-2-cyclopentene-1-methanol succinate); 2', 3'-dideoxyinosine (DDI);
and 2', 3'-dideoxy-2', 3'-didehydrothymidine (D4T), and esters
thereof.
[0014] In light of the strong activity of MKC-442 against HIV, it
is an object of the present invention to provide a method and
composition that includes MKC-442 for the treatment of patients
infected with HIV that exhibits advantageous or improved
pharmacokinetic, biodistribution, metabolic, resistance or other
parameters over administration of MKC-442 alone.
[0015] It is also an object of the invention to improve the
efficacy of MKC-442 during short periods of administration and over
extended time periods.
[0016] It is yet another object of the present invention to provide
a method and composition for the treatment of patients infected
with HIV in which MKC-442 is administered in combination or
alternation with a second compound that acts synergistically with
MKC-442 against the virus.
[0017] It is still another object of the present invention to
provide a method and composition for the treatment of patients
infected with HIV in which MKC-442 is administered in combination
or alternation with a second compound (or at least one other
compound) that acts synergistically with MKC-442 against the
virus.
SUMMARY OF THE INVENTION
[0018] It has been discovered that MKC-442 can be administered in
combination with one or more antiviral agents to achieve an
advantageous therapeutic effect against HIV. In some cases, the
enhanced therapeutic effect is not attainable by administration of
either agent alone. In a preferred but not necessary embodiment,
the effect of administration of the two agents in combination or
alternation is synergistic.
[0019] In one preferred embodiment, MKC-442 is administered in
combination with a protease inhibitor. In particular embodiments,
MKC-442 is administered in combination or alternation with either
indinavir, nelfinavir
([3S-[2(2S*,3S*),3-alpha,4-a-beta,8a-beta-]]-N-(1,1-dimethylet-
hyl)decahydro-2-)2-hydroxy-3-[(3-hydroxy-2-methylbenzoyl)amino]-4-(phenylt-
hio)butyl]-3-isoquinolincarboxamide mono-methanesulfonate),
saquinavir, or amprenavir (141W94) (S)-tetrahydrofuran-3-yl-N-[(1S,
2R)-3-[N-[(4-aminophenyl)-sulfonyl]-N-isobutylamino]-1-benzyl-2-hydroxypr-
opyl]carbamate.
[0020] In another preferred embodiment, MKC-442 is administered in
combination or alternation with a nucleoside analog such as
abacavir (1592U89) which is
(1S,4R)-4-[2-amino-6-cyclopropyl-amino)-9H-purin-9-yl]-
-2-cyclopentene-1-methanol succinate, 3TC, or FTC.
[0021] In another embodiment, MKC-442 is administered in
combination with a non-nucleoside reverse transcriptase inhibitor,
such as DMP-266 (efavirenz;
(S)-6-chloro-4-(cyclopropylethynyl)-1,4-dihydro-4-(trifluorom-
ethyl)-2H-3, 1-benzoxazin-2-one), delavirdine
(1-[3-(1-methyl-ethyl)amino]-
-2-pyridinyl-4-[[5-[(methylsulfonyl)amino]-
1H-indol-2-yl]carbonyl]-, monoethanesulfonate), or nevirapine.
[0022] In general, during alternation therapy, an effective dosage
of each agent is administered serially, whereas in combination
therapy, an effective dosage of two or more agents are administered
together. The dosages will depend on such factors as absorption,
biodistribution, metabolism and excretion rates for each drug as
well as other factors known to those of skill in the art. It is to
be noted that dosage values will also vary with the severity of the
condition to be alleviated. It is to be further understood that for
any particular subject, specific dosage regimens and schedules
should be adjusted over time according to the individual need and
the professional judgment of the person administering or
supervising the administration of the compositions. Examples of
suitable dosage ranges for protease inhibitors, including for
example, nelfinavir and indinavir, can be found in the scientific
literature and in the Physicians Desk Reference. Many examples of
suitable dosage ranges for other compounds described herein are
found in public literature or can be determined easily using known
methods. These dosage ranges can be modified as desired to achieve
a desired result.
[0023] The disclosed combination and alternation regiments are
useful in the prevention and treatment of HIV infections and other
related conditions such as AIDS-related complex (ARC), persistent
generalized lymphadenopathy (PGL), AIDS-related neurological
conditions, anti-HIV antibody positive and HIV-positive conditions,
Kaposi's sarcoma, thrombocytopenia purpurea and opportunistic
infections. In addition, these compounds or formulations can be
used prophylactically to prevent or retard the progression of
clinical illness in individuals who are anti-HIV antibody or
HIV-antigen positive or who have been exposed to HIV.
BRIEF DESCRIPTION OF THE FIGURES
[0024] FIG. 1 is an isobologram analysis of the combination of
MKC-442 and indinavir in cell culture.
[0025] FIG. 2 is an isobologram analysis of the combination of
MKC-442 and nelfinavir in cell culture.
DETAILED DESCRIPTION OF THE INVENTION
[0026] It has been discovered that MKC-442 can be administered in
combination with one or more antiviral agents to achieve an
advantageous therapeutic effect to inhibit HIV replication. In most
cases, the enhanced therapeutic effect is not attainable by
administration of either agent alone. In a preferred but not
necessary embodiment, the effect of administration of the two
agents in combination or alternation is synergistic.
[0027] I. MKC-442 and Related Compounds
[0028] In one embodiment an effective HIV-treatment amount of a
compound of the formula: 2
[0029] wherein X is oxygen or sulfur, R.sup.1 is ethyl or
isopropyl, R.sup.2 and R.sup.3 are independently hydrogen,
C.sub.1-C.sub.4 alkyl or halogen (chlorine, bromine, iodine, or
fluorine) is administered to a patient in need thereof in
combination with a protease inhibitor or with another anti-HIV
compound as described herein. In a preferred embodiment, the
compound is MKC-442.
[0030] Any of the compounds described herein for combination or
alternation therapy can be administered as any derivative that upon
administration to the recipient, is capable of providing directly
or indirectly, the parent compound, or that exhibits activity
itself. Nonlimiting examples are the pharmaceutically acceptable
salts (alternatively referred to as "physiologically acceptable
salts"), and a compound which has been alkylated or acylated at an
appropriate position. The modifications can affect the biological
activity of the compound, in some cases increasing the activity
over the parent compound. This can easily be assessed by preparing
the derivative and testing its antiviral activity according to
known methods.
[0031] As used herein, the term pharmaceutically acceptable salts
refers to salts that retain the desired biological activity of the
herein-identified compounds and exhibit minimal undesired
toxicological effects. Non-limiting examples of such salts are (a)
acid addition salts formed with inorganic acids (for example,
hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric
acid, nitric acid, and the like), and salts formed with organic
acids such as amino acid, acetic acid, oxalic acid, tartaric acid,
succinic acid, malic acid, ascorbic acid, benzoic acid, tannic
acid, palmoic acid, alginic acid, polyglutamic acid,
naphthalenesulfonic acid, naphthalenedisulfonic acid, and
polygalacturonic acid; (b) base addition salts formed with metal
cations such as zinc, calcium, bismuth, barium, magnesium,
aluminum, copper, cobalt, nickel, cadmium, sodium, potassium, and
the like, or with a cation formed from ammonia,
N,N-dibenzylethylene-diamine, D-glucosamine, tetraethylammonium, or
ethylenediamine; or (c) combinations of (a) and (b); e.g., a zinc
tannate salt or the like.
[0032] II. Combination or Alternation Therapy
[0033] It has been recognized that drug-resistant variants of HIV
can emerge after prolonged treatment with an antiviral agent. Drug
resistance most typically occurs by mutation of a gene that encodes
for an enzyme used in the viral life cycle, and most typically in
the case of HIV, in either the reverse transcriptase or protease
genes. It has been demonstrated that the efficacy of a drug against
HIV infection can be prolonged, augmented, or restored by
administering the compound in combination or alternation with a
second, and perhaps third, antiviral compound that induces a
different mutation(s) from that selected for by the principle drug.
Alternatively, the pharmacokinetics, biodistribution, or other
parameter of the drug can be altered by such combination or
alternation therapy. In general, combination therapy is typically
preferred over alternation therapy because it induces multiple
simultaneous stresses on the virus.
[0034] The second antiviral agent for the treatment of HIV, in one
embodiment, can be a protease inhibitor, or a reverse transcriptase
inhibitor (a "RTI"), which can be either a synthetic nucleoside-(a
"NRTI") or a non-nucleoside compound (a "NNRTI"). In other
embodiments, the second (or third) compound can be a pyrophosphate
analog, or a fusion binding inhibitor. A list compiling resistance
data collected in vitro and in vivo for a number of antiviral
compounds is found in Schinazi et al., mutations in retroviral
genes associated with drug resistance, International Antiviral
News, Volume 1 (4), International Medical Press 1996.
[0035] In preferred embodiments, MKC-442 is administered in
combination or alternation with FTC (2',
3'-dideoxy-3'-thia-5-fluorocytidine); 141W94 (amprenavir, Glaxo
Wellcome, Inc.); Viramune (nevirapine), Rescriptor (delavirdine);
or DMP-266 (efavirenz). In another preferred embodiment, MKC-442 is
administered in combination or alternation with abacavir (1592U89),
which is (1S,4R)-4-[2-amino-6-cyclopropyl-amino)-9H-purin-9-yl-
]-2-cyclopentene-1-methanol succinate.
[0036] Other examples of antiviral agents that can be used in
combination or alternation with the compounds disclosed herein for
HIV therapy include carbovir, acyclovir, interferon, stavudine,
CS-92 (3'-azido-2', 3'-dideoxy-5-methyl-cytidine), and
.beta.-D-dioxolane nucleosides such as .beta.-D-dioxolanylguanine
(DXG), .beta.-D-dioxolanyl-2,6-diaminopurine (DAPD), and
.beta.-D-dioxolanyl-6-chloropurine (ACP).
[0037] Preferred protease inhibitors include indinavir
({1(1,S,2R),5(S)]-2,3,5-trideoxy-N-(2,3-dihydro-2-hydroxy-1H-inden-1-yl)--
5-[2-[[(1,1-dimethylethyl)amino]carbonyl]-4-(3-pyridinylmethyl)-1-piperazi-
nyl]-2-(phenylmethyl)-D-erythro-pentoamide sulfate; Merck),
nelfinavir (Agouron), ritonavir (Abbot), and saquinavir (Invirase;
Roche).
[0038] Nonlimiting examples of other compounds that can be
administered in combination or alternation with MKC-442 to augment
the properties of the drug on administration include abacavir:
(1S,4R)-4-[2-amino-6-cyclopropyl-
-amino)-9H-purin-9-yl]-2-cyclopentene-1-methanol succinate
(1592U89, a carbovir analog; Glaxo Wellcome); AzddU:3'-azido-2',
3'-dideoxyuridine; zidovudine: AZT, 3'-azido-3'-deoxythymidine
(Glaxo Wellcome); BILA 1906: N-{1
S-[[[3-[2S-{(1,1-dimethylethyl)amino]carbonyl}-4R-]3-pyridinylmethyl-
)thio]-1-piperidinyl]-2R-hydroxy-1S-(phenylmethyl)propyl]amino]carbonyl]-2-
-methylpropyl}-2-quinolinecarboxamide (Bio
Mega/Boehringer-Ingelheim); BILA 2185:
N-(1,1-dimethylethyl)-1-[2S-[[2-2,6-dimethylphenoxy)-1-oxoethy-
l]amino]-2R-hydroxy-4-phenylbutyl]4R-pyridinylthio)-2-piperidinecarboxamid-
e (Bio Mega/Boehringer-Ingelheim); BM+51.0836:triazoloisoindolinone
derivative; BMS 186,318: aminodiol derivative HIV-1 protease
inhibitor (Bristol-Myers-Squibb); d4API:
9-[2,5-dihydro-5-(phosphonomethoxy)-2-fura- nel]adenine (Gilead);
stavudine: d4T, 2', 3'-didehydro-3'-deoxythymidine
(Bristol-Myers-Squibb); efavirenz: DMP-266, a 1,4-dihydro-2H-3,
1-benzoxazin-2-one; HBY097:
S-4-isopropoxycarbonyl-6-methoxy-3-(methylthi-
o-methyl)-3,4-dihydroquinoxalin-2(1 H)-thione; HEPT:
1-[(2-hydroxyethoxy)methyl]6-(phenylthio)thymine; KNI-272:
(2S,3S)-3-amino-2-hydroxy-4-phenylbutyric acid-containing
tripeptide; L-697,593;
5-ethyl-6-methyl-3-(2-phthalimido-ethyl)pyridin-2(1H)-one;
L-735,524: hydroxy-aminopentane amide HIV-1 protease inhibitor
(Merck); L-697,661:
3-{[(-4,7-dichloro-1,3-benzoxazol-2-yl)methyl]amino}-5-ethyl-6-
-methylpyridin-2(1H)-one; L-FDDC: (-)-.beta.-L-5-fluoro-2',
3'-dideoxycytidine; L-FDOC: (-)-.beta.-L-5-fluoro-dioxolane
cytosine; 6-benzyl-1-ethoxymethyl-5-isopropyluracil (I-EBU;
Triangle/Mitsubishi); nevirapine:
11-cyclopropyl-5,11-dihydro-4-methyl-6H-dipyridol[3,2-b:2',
phosphonoformate (foscarnet; Astra); PMEA:
9-(2-phosphonylmethoxyethyl) adenine (Gilead); PMPA:
(R)-9-(2-phosphonyl-methoxypropyl)adenine (Gilead); Ro 31-8959:
hydroxythethylamine derivative HIV-1 protease inhibitor (Roche);
RPI-3121: peptidyl protease inhibitor,
1-[(3s)-3-(n-alpha-benzyloxycarbonyl)-1-asparginyl)-amino-2-hydroxy-4-phe-
nylbutyryl]-n-tert-butyl-1-proline amide; 2720:
6-chloro-3,3-dimethyl-4-(i-
sopropenyloxycarbonyl)-3,4-dihydro-quinoxalin-2(1H)thione;
SC-52151: hydroxyethylurea isostere protease inhibitor (Searle);
SC-55389A: hydroxyethyl-urea isostere protease inhibitor (Searle);
TIBO R82150:
(+)-(5S)-4,5,6,7-tetrahydro-5-methyl-6-(3-methyl-2-butenyl)imidazo[4,5,1--
jk][1,4]-benzodiazepin-2(1H)-thione (Janssen); TIBO 82913:
(+)-(5S)-4,5,6,7,-tetrahydro-9-chloro-5-methyl-6-(3-methyl-2-butenyl)imid-
azo[4,5,ljk]-[1,4]benzodiazepin-2(1H)-thione (Janssen); TSAO-m3T:
[2', 5'-bis-O-(tert-butyldimethyls N3-methylthymine;
U90152:1-[3-[(1-methyleth-
yl)-amino]2-pyridinyl]-4-[[5-[(methylsulphonyl)-amino]-1H-indol-2yl]carbon-
yl]piperazine; UC: thiocarboxanilide derivatives (Uniroyal); UC-781
=N-[4-chloro-3-(3-methyl-2-butenyloxy)phenyl]-2-methyl-3-furancarbothioam-
ide; UC-82
=N-[4-chloro-3-(3-methyl-2-butenyloxy)phenyl]-2-methyl-3-thioph-
enecarbothioamide; VB 11,328: hydroxyethylsulphonamide protease
inhibitor (Vertex); VX-478: amprenavir, 141W94,
hydroxyethylsulphonamide protease inhibitor (Vertex/Glaxo
Wellcome); XM 323: cyclic urea protease inhibitor (Dupont Merck),
delaviridine (Pharmacia Upjohn), famciclovir, gancyclovir, and
penciclovir. In another embodiment, MKC-442 is administered in
combination with LG1350, which has the following structure. 3
[0039] Nucleoside Prodrug Formulations
[0040] Any of the nucleosides or other compounds which are
described herein for use in combination or alternation therapy with
MKC-442 or its related compounds can be administered as an acylated
prodrug or a nucleotide prodrug, as described in detail below.
[0041] The term acyl refers to a carboxylic acid ester in which the
non-carbonyl moiety of the ester group is selected from straight,
branched, or cyclic alkyl or lower alkyl, alkoxyalkyl including
methoxymethyl, aralkyl including benzyl, aryloxyalkyl such as
phenoxymethyl, aryl including phenyl optionally substituted with
halogen, C.sub.1 to C.sub.4 alkyl or C.sub.1 to C.sub.4 alkoxy,
sulfonate esters such as alkyl or aralkyl sulphonyl including
methanesulfonyl, the mono, di or triphosphate ester, trityl or
monomethoxytrityl, substituted benzyl, trialkylsilyl (e.g.
dimethyl-t-butylsilyl
[0042] Any of the nucleosides described herein (or NNRTIs) or other
compounds that have a hdyroxyl group, or amine function, can be
administrated as a nucleotide or other prodrug to increase the
activity, bioavailability, stability or otherwise alter the
properties of the nucleoside. A number of nucleotide prodrug
ligands are known. In general, alkylation, acylation or other
lipophilic modification of the mono, di or triphosphate of the
nucleoside will increase the stability of the nucleotide. Examples
of substituent groups that can replace one or more hydrogens on the
phosphate moiety are alkyl, aryl, steroids, carbohydrates,
including sugars, 1,2-diacylglycerol and alcohols. Many are
described in R. Jones and N. Bischofberger, Antiviral Research, 27
(1995) 1-17. Any of these can be used in combination with the
disclosed nucleosides or other compounds to achieve a desire
effect.
[0043] The nucleosides (or NNRTIs) can also be provided as a
5'-ether lipid, as disclosed in the following references, which are
incorporated by reference herein: Kucera, L. S., N. Iyer, E. Leake,
A. Raben, Modest E. K., D. L. W., and C. Piantadosi. 1990. "Novel
membrane-interactive ether lipid analogs that inhibit infectious
HIV-1 production and induce defective virus formation." AIDS Res.
Hum. Retro Viruses. 6:491-501; Piantadosi, C., J. Marasco C. J., S.
L. Morris-Natschke, K. L. Meyer, F. Gumus, J. R. Surles, K. S.
Ishaq, L. S. Kucera, N. Iyer, C. A. Wallen, S. Piantadosi, and E.
J. Modest. 1991. "Synthesis and evaluation of novel ether lipid
nucleoside conjugates for anti-HIV activity." J. Med. Chem.
34:1408.1414; Hosteller, K. Y., D. D. Richman, D. A. Carson, L. M.
Stuhmiller, G. M. T. van Wijk, and H. van den Bosch. 1992. "Greatly
enhanced inhibition of human immunodeficiency virus type 1
replication in CEM and HT4-6C cells by 3'-deoxythymidine
diphosphate dimyristoylglycerol, a lipid prodrug of
3,-deoxythymidine." Antimicrob. Agents Chemother. 36:2025.2029;
Hostetler, K. Y., L. M. Stuhmiller, H. B. Lenting, H. van den
Bosch, and D. D. Richman, 1990. "Synthesis and antiretroviral
activity of phospholipid analogs of azidothymidine and other
antiviral nucleosides." J. Biol. Chem. 265:61127.
[0044] Nonlimiting examples of U.S. patents that disclose suitable
lipophilic substituents that can be covalently incorporated into
the nucleoside or other hydroxyl or amine containing compounds,
preferably at the 5'-OH position of the nucleoside or lipophilic
preparations, include U.S. Pat. Nos. 5,149,794 (Sep. 22, 1992,
Yatvin et al.); 5,194,654 (Mar. 16, 1993, Hostetler et al.,
5,223,263 (Jun. 29, 1993, Hostetler et al.); 5,256,641 (Oct. 26,
1993, Yatvin et al.); 5,411,947 (May 2, 1995, Hostetler et al.);
5,463,092 (Oct. 31, 1995, Hostetler et al.); 5,543,389 (Aug. 6,
1996, Yatvin et al.); 5,543,390 (Aug. 6, 1996, Yatvin et al.);
5,543,391 (Aug. 6, 1996, Yatvin et al.); and 5,554,728 (Sep. 10,
1996; Basava et al.), all of which are incorporated herein by
reference. Foreign patent applications that disclose lipophilic
substituents that can be attached to the nucleosides of the present
invention, or lipophilic preparations, include WO 89/02733, WO
90/00555, WO 91/16920, WO 91/18914, WO 93/00910, WO 94/26273, WO
96/15132, EP 0 350 287, EP 93917054.4, and WO 91/19721.
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[0046] Alkyl hydrogen phosphate derivatives of the anti-HIV agent
AZT may be less toxic than the parent nucleoside analogue.
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[0047] These compounds can be tested for synergistic activity with
MKC-442 against HIV according to a number of assays, including that
described below.
EXAMPLE 1
Evaluation of Inhibition of HIV Infection Using Combined Antiviral
Agents
[0048] The inhibitory effects of combined antiviral agents were
evaluated using the human T cell line, MT2. Infection of this cell
line with HIV results in cell death. Inhibition of the HIV induced
cytopathic effect by the combined agents is measured in a MTT
(3-[4,5-dimethylthiazol-2-yl]-2,- 5-diphenyl tetrazolium bromide)
based assay. The mitochondrial dehydrogenases of viable cells will
cleave the tetrazolium ring of MTT, yielding purple formazan
crystals which are soluble in acidified isopropanol. The resulting
purple solution is measured spectrophotometrically. The amount of
purple dye formed is directly related to the number of viable
cells. Therefore, a decrease in color in this assay indicates cell
death due to viral cytopathic effects (CPE). This assay has been
employed in the assessment of a variety of antiretroviral compounds
(Pauwels, R. et al, 1988, J. Virol. Methods, 20: 309-321.) and is
readily adaptable to any system with a lytic virus.
[0049] Preparation of Drug Plates
[0050] The assays are performed in a 96 well microtiter plate. Each
compound to be tested is made up at 4 times the final concentration
to allow for dilution by cells and other test compounds. Compounds
are prepared in complete RPMI media (RPMI 1640 plus 10% fetal
bovine serum plus 20 mg/mL gentamicin). Two drugs are tested
simultaneously on each plate using a checkerboard design. Serial
dilutions of the first drug are added to the vertical wells (50
mL/well) of the plate (columns 2- 11). Serial dilutions of the
second drug (50 mL/well) are then added to the plate across
horizontal rows (rows A - G). Column #2 and row H contain dilutions
of only one compound. EC.sub.50 values obtained from these wells
are a measure of the antiviral activity of each compound alone.
Column #1 contains virus control, infected cells grown in the
absence of compound, and Column 12 contains cell control, mock
infected cells.
[0051] Preparation of Cells
[0052] MT2 cells are infected with the LAI strain of HIV at a
multiplicity of infection of 0.01 in complete RPMI containing 2
mg/mL polybrene for 2 hours at 37.degree. C. Following infection,
cells diluted to 2.5.times.10.sup.5 cells/mL in complete RPMI
media. 100 mL of the cell suspension are seeded onto 96 well
plates, 2.5.times.10.sup.4 cells/well, containing test compounds as
described above. The final volume in each well is 200 mL. Plates
are sealed and incubated at 37.degree. C. in a humidified 5%
CO.sub.2 incubator for 5 days.
[0053] Developing the Assay
[0054] Following the 5 day incubation, 80 mL of cell free
supenatant is removed from each well and 10 mL of a 5 mg/mL
solution of MTT is added. The plates are returned to the incubator
for 2 hours followed by the addition of 100 mL of acidified
isopropanol (10% Triton X-100 plus 0.1N HCl in anhydrous
isopropanol) to each well. Formazan crystals are dissolved by
repeated pipetting in the wells. Plates are read at 570 nm in a
Biotek plate reader.
[0055] Calculation of Data
[0056] A dose response curve for each individual compound is
generated using the absorption values of the cell controls as 100%
protection and no drug virus infected cells as 0% protection. From
this dose response curve a EC.sub.50 value is calculated and is
defined as the concentration of drug that inhibits 50% of viral
induced CPE. The effects of the combined compounds were obtained
using the isobologram technique (Elion, G. B. et al, 1954, J. Biol.
Chem., 208: 477-488). The data plotted were obtained by dividing
the EC.sub.50 of compound #1 in the presence of a fixed
concentration of compound #2 by the EC.sub.50 of compound #1 alone.
In a similar fashion data can be obtained by dividing the EC.sub.50
of compound #2 in the presence of a fixed concentration of compound
#1 by the EC.sub.50 of compound #2 alone. The EC.sub.50 ratios are
plotted against the ratio of compound at each combination. If the
curve generated by these points falls on the theoretic diagonal
line, the combination is additive. If the curve is above the
theoretical line, the combination is antagonistic. And, if the
curve lies below the theoretical line, then the combination is
synergistic. FIGS. 1 and 2 illustrate isobologram analysis of the
combination of MKC-442 with indinavir and with nelfinavir.
[0057] Even though a combination exhibits additive and not
synergistic effects according to this assay, the combination can
still provide an effect that is different from the separate
administration of the two agents. For example, the biodistribution,
pharmacokinetics, or metabolism of one can be affected by the
other. Further, MKC-442 is advantageously delivered in combination
or alternation with another antiviral agent that exhibits a
different mutation profile than MKC-442.
PREPARATION OF PHARMACEUTICAL COMPOSITIONS
[0058] Humans suffering from effects caused by any of the diseases
described herein, and in particular, HIV infection, can be treated
by administering to the patient an effective amount of MKC-442 in
combination with one or more agents described above or a
pharmaceutically acceptable derivative or salt thereof in the
presence of a pharmaceutically acceptable carrier or diluent. The
active materials can be administered by any appropriate route, for
example, orally, parenterally, enterally, intravenously,
intradermally, subcutaneously, or topically, in liquid or solid
form.
[0059] The active compounds are included in the pharmaceutically
acceptable carrier or diluent in an amount sufficient to deliver to
a patient a therapeutically effective amount of compound to inhibit
viral replication in vivo, especially HIV replication, without
causing serious toxic effects in the treated patient. By
"inhibitory amount" is meant an amount of active ingredient
sufficient to exert an inhibitory effect as measured by, for
example, an assay such as the ones described herein.
[0060] A preferred dose of the compounds for all the
above-mentioned conditions will be in the range from about 1 to 50
mg/kg, preferably 1 to 20 mg/kg, of body weight per day, more
generally 0.1 to about 100 mg per kilogram body weight of the
recipient per day. The effective dosage range of the
pharmaceutically acceptable derivatives can be calculated based on
the weight of the parent nucleoside to be delivered. If the
derivative exhibits activity in itself, the effective dosage can be
estimated as above using the weight of the derivative, or by other
means known to those skilled in the art.
[0061] The compounds are conveniently administered in unit any
suitable dosage form, including but not limited to one containing 7
to 3000 mg, preferably 70 to 1400 mg of active ingredient per unit
dosage form. An oral dosage of 50 to 1000 mg is usually
convenient.
[0062] Ideally, the active ingredients should be administered to
achieve peak plasma concentrations of the active compound of from
about 0.2 to 70 .mu.M, preferably about 1.0 to 10 .mu.M. This may
be achieved, for example, by the intravenous injection of a 0.1 to
% solution of the active ingredient, optionally in saline, or
administered as a bolus of the active ingredient.
[0063] The concentration of active compound in the drug composition
will depend on absorption, distribution, metabolism and excretion
rates of the drug as well as other factors known to those of skill
in the art. It is to be noted that dosage values will also vary
with the severity of the condition to be alleviated. It is to be
further understood that for any particular subject, specific dosage
regimens should be adjusted over time according to the individual
need and the professional judgment of the person administering or
supervising the administration of the compositions, and that the
concentration ranges set forth herein are exemplary only and are
not intended to limit the scope or practice of the claimed
composition. The active ingredient may be administered at once, or
may be divided into a number of smaller doses to be administered at
varying intervals of time.
[0064] A preferred mode of administration of the active compound is
oral. Oral compositions will generally include an inert diluent or
an edible carrier. They may be enclosed in gelatin capsules or
compressed into tablets. For the purpose of oral therapeutic
administration, the active compound can be incorporated with
excipients and used in the form of tablets, troches, or capsules.
Pharmaceutically compatible bind agents, and/or adjuvant materials
can be included as part of the composition.
[0065] The tablets, pills, capsules, troches and the like can
contain any of the following ingredients, or compounds of a similar
nature: a binder such as microcrystalline cellulose, gun tragacanth
or gelatin; an excipient such as starch or lactose, a
disintegrating agent such as alginic acid, Primogel, or corn
starch; a lubricant such as magnesium stearate or Sterotes; a
glidant such as colloidal silicon dioxide; a sweetening agent such
as sucrose or saccharin; or a flavoring agent such as peppermint,
methyl salicylate, or orange flavoring. When the dosage unit form
is a capsule, it can contain, in addition to material of the above
type, a liquid carrier such as a fatty oil. In addition, dosage
unit forms can contain various other materials which modify the
physical form of the dosage unit, for example, coatings of sugar,
shellac, or other enteric agents.
[0066] The compounds can be administered as a component of an
elixir, suspension, syrup, wafer, chewing gum or the like. A syrup
may contain, in addition to the active compounds, sucrose as a
sweetening agent and certain preservatives, dyes and colorings and
flavors.
[0067] The compounds or their pharmaceutically acceptable
derivative or salts thereof can also be mixed with other active
materials that do not impair the desired action, or with materials
that supplement the desired action, such as antibiotics,
antifungals, antiinflammatories, protease inhibitors, or other
nucleoside or non-nucleoside antiviral agents, as discussed in more
detail above. Solutions or suspensions used for parental,
intradermal, subcutaneous, or topical application can include the
following components: a sterile diluent such as water for
injection, saline solution, fixed oils, polyethylene glycols,
glycerine, propylene glycol or other synthetic solvents;
antibacterial agents such as benzyl alcohol or methyl parabens;
antioxidants such as ascorbic acid or sodium bisulfite; chelating
agents such as ethylenediaminetetraacetic acid; buffers such as
acetates, citrates or phosphates and agents for the adjustment of
tonicity such as sodium chloride or dextrose. The parental
preparation can be enclosed in ampoules, disposable syringes or
multiple dose vials made of glass or plastic.
[0068] If administered intravenously, preferred carriers are
physiological saline or phosphate buffered saline (PBS).
[0069] In a preferred embodiment, the active compounds are prepared
with carriers that will protect the compound against rapid
elimination from the body, such as a controlled release
formulation, including implants and microencapsulated delivery
systems. Biodegradable, biocompatible polymers can be used, such as
ethylene vinyl acetate, polyanydrides, polyglycolic acid, collagen,
polyorthoesters, and polylactic acid. Methods for preparation of
such formulations will be apparent to those skilled in the art. The
materials can also be obtained commercially from Alza
Corporation.
[0070] Liposomal suspensions (including liposomes targeted to
infected cells with monclonal antibodies to viral antigens) are
also preferred as pharmaceutically acceptable carriers. these may
be prepared according to methods known to those skilled in the art,
for example, as described in U.S. Pat. No. 4,522,811 (which is
incorporated herein by reference in its entirety). For example,
liposome formulations may be prepared by dissolving appropriate
lipid(s) (such as stearoyl phosphatidyl ethanolamine, stearoyl
phosphatidyl choline, arachadoyl phosphatidyl choline, and
cholesterol) in an inorganic solvent that is then evaporated,
leaving behind a thin film of dried lipid on the surface of the
container. An aqueous solution of the active compound or its
monophosphate, diphosphate, and/or triphosphate derivatives is then
introduced into the container. The container is then swirled by
hand to free lipid material from the sides of the container and to
disperse lipid aggregates, thereby forming the liposomal
suspension.
[0071] This invention has been described with reference to its
preferred embodiments. Variations and modifications of the
invention, will be obvious to those skilled in the art from the
foregoing detailed description of the invention. It is intended
that all of these variations and modifications be included within
the scope of this invention.
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