WO1997003055A1 - Composes, compositions et procedes pour inhiber la replication de retrovirus et pour inhiber une transcription induite par un promoteur de tumeur - Google Patents

Composes, compositions et procedes pour inhiber la replication de retrovirus et pour inhiber une transcription induite par un promoteur de tumeur Download PDF

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Publication number
WO1997003055A1
WO1997003055A1 PCT/US1996/011699 US9611699W WO9703055A1 WO 1997003055 A1 WO1997003055 A1 WO 1997003055A1 US 9611699 W US9611699 W US 9611699W WO 9703055 A1 WO9703055 A1 WO 9703055A1
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oltipraz
composition
subject
replication
retrovirus
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PCT/US1996/011699
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English (en)
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Hans J. Prochaska
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Sloan-Kettering Institute For Cancer Research
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Priority to AU66767/96A priority Critical patent/AU6676796A/en
Publication of WO1997003055A1 publication Critical patent/WO1997003055A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/4985—Pyrazines or piperazines ortho- or peri-condensed with heterocyclic ring systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/437—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a five-membered ring having nitrogen as a ring hetero atom, e.g. indolizine, beta-carboline
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/55—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/55—Protease inhibitors
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
    • C07D409/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings
    • C07D409/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems

Definitions

  • COMPOUNDS COMPOSITIONS, AND METHODS FOR INHIBITING REPLICATION OF RETROVIRUSES AND FOR INHIBITING TUMOR PROMOTER INITIATED TRANSCRIPTION
  • Oltipraz (5-pyrazinyl-4-methyl-l, 2-dithiole-3-thione) , which was developed and tested in humans as an antischistosomal agent, is a highly effective inhibitor of chemical carcinogenesis (1-3) .
  • the effectiveness of oltipraz in preventing neoplasia in a wide variety of experimental models has prompted a clinical evaluation of oltipraz as a potential human anticarcinogen (4-8) .
  • Oltipraz is also an inhibitor of HIV-1 replication (9) .
  • RT reverse transcriptase
  • RT reverse transcriptase
  • oltipraz also inhibits viral replication in a chronic infection model (10) , oltipraz appears to possess an additional antiviral mechanism.
  • the potency of the drug in Ul cells is markedly potentiated by preincubation of the cells with oltipraz before PMA stimulation (10) .
  • This invention involves the novel notion that metabolites of oltipraz may also play a role in the inhibition of viral replication.
  • the metabolic fate of oltipraz is complex (7, 8, 11, 12) . Less than 1% of the administered dose is excreted unaltered in the urine.
  • the major metabolic pathway for oltipraz is likely to occur via a scenario involving opening of the 1, 2-dithiole- 3-thione ring by a nucleophile (perhaps glutathione) , followed by a two-electron reduction of the adduct, elimination of the nucleophile, and recyclization (13) .
  • the resulting species is then methylated to form 7-methyl-6-8-bis (methylthio) pyrrolo [1,2-a] pyrazine.
  • ACH-2 cells produce low levels of infectious virus under basal conditions but HIV-1 replication can be markedly induced with PMA (14) .
  • CEM cells were used in the experiments described below not only because they represent another widely utilized model for acute viral replication in T cells but also because they are progenitors of A3.01 cells, from which the ACH-2 cell line was derived. Thus, it would be expected that differences in metabolism and uptake of oltipraz would be minimized between CEM and ACH-2 cells, rendering more valid a comparison of oltipraz and metabolite III in models of acute versus chronic viral replication.
  • PBMCs were tested in the experiments described below because they are the closest model of in vivo conditions. This invention involves the discovery, established by the experiments described below, that metabolite III possesses antiviral activity at a step distal to viral integration and is synergistic with the parent compound (oltipraz) .
  • Oltipraz which was developed clinically as an antischistosomal agent, has received intensive scrutiny since it is a highly effective inhibitor of experimental carcinogenesis.
  • the anticarcinogenic activity of oltipraz was predicted by the ability of the drug to induce Phase II detoxication enzymes (i.e., GSH transferases) and enzymes concerned with the synthesis and maintenance of GSH pool (1, 25) .
  • Phase II detoxication enzymes i.e., GSH transferases
  • the induction of Phase II enzymes appears to be relevant to its ability to prevent cancer in a number of experimental models.
  • oltipraz is a highly effective inhibitor of aflatoxin Bj-induced hepatocarcinogenesis in the F344 Fischer rat (2, 22), and these results have prompted a Phase II clinical trial in Qidong, China to ascertain if the drug can reduce urinary levels of DNA-aflatoxin adducts (26) .
  • the chemopreventive activity of oltipraz can be attributed to the induction of Phase II enzymes under many circumstances, the mechanism for protection is less clear in others.
  • oltipraz is as effective as an inhibitor of tumor promotion as it is in the initiation phase of azoxymethane (AOM) -induced colon carcinogenesis.
  • AOM azoxymethane
  • oltipraz has also been determined to inhibit HIV-1 replication in vi tro (9) .
  • Oltipraz may act as an antiretroviral by irreversibly inhibiting HIV-1 RT in acute infection models, however the finding that the drug possesses antiretroviral activity in chronically infected Ul promonocytic leukemia cells led to the conclusion that oltipraz exerts another antiviral mechanism of action (10) .
  • the metabolism of oltipraz is complex with the generation of series of rearranged metabolites that are formed from 7-methyl-6, 8-bis (methylthio)pyrrolo- [1, 2-a] - pyrazine (metabolite III;) (11, 12) .
  • Most of the rearranged metabolites differ from metabolite III primarily in the oxidation state of the pyrazine ring or sulfur atoms.
  • This invention provides a pharmaceutical composition for inhibiting replication of a retrovirus in a subject infected with the retrovirus which comprises a pharmaceutically acceptable carrier and a compound having the structure:
  • Z j is either C-R 5 or N; wherein Z 2 is N; wherein X j and X 2 are the same or different and are O, S, or N-R 4 ; and wherein Ri , R 2 , R 3 , R 4 , and R 5 are the same or different and are either hydrogen or a branched or unbranched alkyl group of from 1 to 3 carbons.
  • This invention also provides a pharmaceutical composition for inhibiting replication of a retrovirus in a subject infected with the retrovirus which comprises a pharmaceutically acceptable carrier, a first compound which inhibits replication of the retrovirus and a second compound having the structure:
  • Zj is either C-R 5 or N; wherein Z 2 is N; wherein X ⁇ and X 2 are the same or different and are 0, S, or N-R 4 ; and wherein R,, R 2 , R 3 , R 4 , and R 5 are the same or different and are either hydrogen or a branched or unbranched alkyl group of from 1 to 3 carbons.
  • Z j is either C-R 5 or N; wherein Z 2 is N; wherein Xj and X 2 are the same or different and are O, S, or N-R 4 ; and wherein Rj, R 2 , R 3 , R 4 , and R 5 are the same or different and are either hydrogen or a branched or unbranched alkyl group of from 1 to 3 carbons.
  • This invention further provides a method of inhibiting replication of a retrovirus in subject infected with the retrovirus which comprises administering to the subject in an amount effective to inhibit retroviral replication the above- described composition which comprises a pharmaceutically acceptable carrier, a first compound which inhibits replication of the retrovirus and a second compound having the structure:
  • Z j is either C-R 5 or N; wherein Z 2 is N; wherein X j and X 2 are the same or different and are 0, S, or N-R 4 ; and wherein R l f R 2 , R 3 , R 4 , and R 5 are the same or different and are either hydrogen or a branched or unbranched alkyl group of from 1 to 3 carbons.
  • This invention further provides a pharmaceutical composition for inhibiting tumor-promoter initiated transcription which comprises a pharmaceutically acceptable carrier and a compound having the structure:
  • Z is either C-R 5 or N; wherein Z 2 is N; wherein X j and X 2 are the same or different and are 0, S, or N-R 4 ; and wherein R,, R 2 , R 3 , R 4 , and R 5 are the same or different and are either hydrogen or a branched or unbranched alkyl group of from 1 to 3 carbons.
  • Z j is either C-R 5 or N; wherein Z 2 is N; wherein X j and X 2 are the same or different and are O, S, or N-R 4 ; and wherein Ri , R 2 , R 3 , R 4 , and R 5 are the same or different and are either hydrogen or a branched or unbranched alkyl group of from 1 to 3 carbons.
  • This invention further describes a compound having the structure:
  • Z j is either C-R 5 or N; wherein Z 2 is N; wherein X ! and X 2 are the same or different and are 0, S, or N-R 4 ; and wherein Rj, R 2 , R 3 , R 4 , and R 5 are the same or different and are either hydrogen or a branched or unbranched alkyl group of from 1 to 3 carbons; with the proviso that when Zj and Z 2 are both N, X j and X 2 are both S, and Rj and R 2 are both methyl, R 3 is not methyl.
  • Fig. 1 Proposed routes for the biotransformation of oltipraz to metabolite III. The scheme was adapted from the work of Fleury et al . (13) . "Nuc" is nucleophile.
  • Fig. 2 Inhibition of p24 antigen release into culture supernatants of H9 (upper) and CEM (lower) cell lines as a function of the concentration of metabolite III or oltipraz.
  • the inhibition of p24 antigen release was determined by normalizing p24 antigen levels to those of matched, HIV-1-infected (drug-free) , control cells. Culture conditions and assays were as described in Experimental Procedures. Symbols, mean + standard error of the percentage inhibition of p24 antigen release for seven and five independent experiments for the H9 and CEM cell lines, respectively.
  • the IC 50 values for zidovudine were determined to be 0.050 and 0.010 ⁇ M for the H9 and CEM cell lines, respectively.
  • Fig. 3 Effects of oltipraz and metabolite III on RT activity as a function of concentration (upper) and the effect of metabolite III as an irreversible inhibitor of RT (lower) .
  • Upper, 10- ⁇ l aliquots of PBS containing RT, 1 mg/ml bovine serum albumin, 3.5% Triton X-100, and 0-525 ⁇ M metabolite III were added to 25 ⁇ l of RT reaction buffer (50 mM Tris- HCL, 75mM KCI, 5 mM MgCl 2 , 1 mM dithiothreitol, 1 mM
  • EGTA 0.1% Nonidet P-40, 0.1% Triton X-100, 10 ⁇ g/ml poly(A) + RNA, 2.5 ⁇ g/ml oligo(dT) , 4 ⁇ M dTTP, 20 ⁇ Ci/ml [ ⁇ - 32 P]dTTP, pH 7,8] in well of 96-well microtiter plate. After the mixtures were incubated for 0.5 hour at 37 ° , the remainder of the isotopic assay was performed as described by Chavan and Prochaska (16) . Symbols , mean ⁇ standard error for the enzyme assay.
  • RT dissolved in PBS containing 1 mg/ml bovine serum albumin was preincubated with 56 (x) , 28 ( ⁇ ) , 14(D) , or 0(O) ⁇ M metabolite III for the times indicated. After incubation, 10- ⁇ l aliquots (in triplicate) were removed and assayed for RT activity as described above.
  • Fig. 4 Inhibition of p24 antigen release into culture supernatants from PMA-stimulated ACH-2 cells by metabolite III (M III) and interferon- ⁇ (IFN) but not oltipraz (OLT) .
  • ACH-2 cells (50,000 cells/microtiter well, 200- ⁇ l final volume) were incubated with various concentrations of oltipraz or metabolite III for 24 hours before the addition of 2.5 nM PMA, in triplicate sets of wells. The cells were grown for an additional 48 hours, and p24 antigen release into culture supernatants was determined. Shown are the means ( ⁇ ) , standard errors (D) , and p values (numbers on the right) ; p values of ⁇ 0.05 were considered significantly different from PMA-stimulated controls.
  • P stimulated lymphocytes were prepared and treated with oltipraz and/or metabolite III and virus as described in the "Experimental Details" Section. Symbols , mean ⁇ standard error for five independent experiments (wherein the combination of oltipraz and metabolite II was tested) . Inset, corresponding combination index as a function of fractional inhibition of p24 antigen release. The median-effect plots [-log [ ( 1/fractional inhibition) -1] as a function of log
  • DMSO dimethyl sulfoxide
  • Fig. 8 Potential mechanisms whereby oltipraz and metabolite III may affect differing stages of the viral life-cycle and carcinogenesis.
  • oltipraz inhibits HIV-1 replication in acute T-cell infection models, and oltipraz inactivates HIV-1 RT 2 ' 3 . This is the likely antiviral mechanism for oltipraz since HIV-1 replication is not blocked by oltipraz treatment in chronically infected ACH-2 T- cell lymphoma cells 1 .
  • Metabolite III affects LTR- driven transcription, and can block viral replication in chronically infected cells.
  • oltipraz can inhibit HIV-1 the initiation phase of carcinogenesis by diverting ultimate carcinogens form interacting with DNA (by induction of Phase II enzymes 4,5 ' 8 and the inhibition of Phase I enzymatic activities 6 ) .
  • Metabolite III is proposed to block the promotion phase of carcinogenesis by blocking cellular proliferations by mitogens and cytokines. This scheme implies that oltipraz may be a more effective antiviral/anticarcinogen than can be appreciated by considering the effects of oltipraz or metabolite III alone.
  • This invention provides a pharmaceutical composition for inhibiting replication of a retrovirus in a subject infected with the retrovirus which comprises a pharmaceutically acceptable carrier and a compound having the structure:
  • R. , R 2 , R 3 , R 4 , and R 5 are the same or different and are either hydrogen or a branched or unbranched alkyl group of from 1 to 3 carbons.
  • Zj and Z 2 are both N; X j and X 2 are both S; and R 1# R 2 , and R 3 are each methyl, i.e. the compound of the composition is 7-methyl-6, 8 bis (methylthio)pyrrolo[1, 2-a]pyrazine, which is sometimes referred to as "metabolite III".
  • the compound of the above composition is useful for inhibiting the replication of any retrovirus, including, but not limited to HIV-1 (Human Immunodeficiency Virus Type 1) , HIV-2 (Human Immunodeficiency Virus Type 2) , HTLV-1 (Human T-Cell Leukemia Virus Type 1) , FIV (Feline Immunodeficiency Virus) , or FLV (Feline Leukemia Virus) .
  • the compound appears to be able to block a transcription factor from binding to an LTR (Long Terminal Repeat) sequence of such retroviruses, and thereby inhibits the activation of transcription of the viral genome of these retroviruses.
  • the composition of this invention is also useful for inhibiting the replication of any virus in which this transcription factor plays a role in the virus's life-cycle, i.e. in the metabolic steps which comprise replication of the virus.
  • the subject in which the above composition inhibits replication of a retrovirus is a mammal.
  • the subject may be any mammal.
  • the subject is a human, in which case the composition of the invention particularly inhibits in the subject replication of retroviruses which most commonly infect humans, such as HIV-1 or HIV-2, or HTLV-I.
  • retroviruses which most commonly infect humans such as HIV-1 or HIV-2, or HTLV-I.
  • the composition would still be useful for inhibiting replication of that retrovirus.
  • the subject is a cat, in which case the composition of the invention particularly inhibits replication of retroviruses which most commonly infect cats, including, but not limited to, FIV or FLV.
  • the composition of the invention also inhibits in cats the replication of retroviruses which do not commonly infect cats.
  • the term "pharmaceutically acceptable carrier” encompasses any of the standard pharmaceutically accepted carriers known to those of ordinary skill in the art. Examples of such standard carriers include, but are not limited to, phosphate buffered saline solution, water, emulsions such as oil/water emulsions or a triglyceride emulsion, various types of wetting agents, tablets, coated tablets and capsules.
  • a suitable pharmaceutically acceptable carrier may be selected taking into account factors known to those of ordinary skill in the art, for example the mode of administration by which the composition is to be administered to the subject.
  • This invention also provides a pharmaceutical composition for inhibiting replication of a retrovirus in a subject infected with the retrovirus which comprises a pharmaceutically acceptable carrier, a first compound which inhibits replication of the retrovirus and a second compound having the structure:
  • Zj is either C-R 5 or N; wherein Z 2 is N; wherein X ] and X 2 are the same or different and are O, S, or N-R 4 ; and wherein R lf R 2 , R 3 , R 4 , and R 5 are the same or different and are either hydrogen or a branched or unbranched alkyl group of from 1 to 3 carbons.
  • Z j and Z 2 of the second compound are both N; Xj and X 2 are both S; and Ri, R 2 , and R 3 are each methyl.
  • this compound is sometimes referred to as "metabolite III”.
  • the first compound of the above-described composition may be any compound which is known to inhibit retroviral replication. Some of the resulting compositions would comprise a * combination of a first compound and a second compound which would be expected to provide a synergistic inhibition of retroviral replication. Accordingly, the subject invention also includes synergistic compositions according to the above- described composition comprising a first compound which inhibits retroviral replication and a second compound having the above-defined structure.
  • the compound in the above-described composition appears to be able to block a transcription factor from binding to the LTR (Long Terminal Repeat) sequences of retroviruses, and thereby inhibits activation of transcription of the integrated viral genome of these retroviruses.
  • the compound of the above-described composition therefore inhibits retrovirus transcription by blocking the transcription activation step in the life-cycle of retroviruses.
  • the first compound inhibits retroviral replication by interfering with a step in the life-cycle of retroviruses other than the transcription activation step.
  • retroviruses Such steps in the life-cycle of retroviruses are well known to those of ordinary skill in the art and include, but are not limited to, binding of the retrovirus to a CD4 receptor on a host cell, internalization of the retrovirus by the host cell, uncoating of the retrovirus, reverse transcription of the retroviral genome, DNA integration into the host cell's genome, and, following transcription activation, production of genomic mRNA, translation of genomic mRNA, processing of retroviral proteins, assembly, and release.
  • the first compound inhibits transcription activation in the retroviral life-cycle. If the first compound may, however, inhibit transcription activation by a mechanism other than by blocking the aforementioned transcription factor from binding to an LTR of the retrovirus.
  • the first compound inhibits protease, an enzyme which processes retroviral proteins during the step of processing retroviral proteins in the retroviral life-cycle.
  • Any compound which inhibits protease may be used in the above- described composition.
  • examples of compounds which inhibit protease include, but are not limited to, saquinavir (made by Hoffman LaRoche) and indinavir sulfate (made by Merck) .
  • the first compound inhibits reverse transcriptase, the enzyme which reverse transcribes the viral genome into DNA during the reverse transcription step of the retroviral life-cycle.
  • Any compound which inhibits reverse transcriptase may be used in the above-described composition.
  • Compounds which inhibit reverse transcriptase include, but are not limited to, AZT
  • composition is useful for inhibiting the replication of any retrovirus, including, but not limited to HIV-1 (Human Immunodeficiency Virus Type 1) , HIV-2 (Human Immunodeficiency Virus Type 2) , HTLV-1 (Human T-Cell Leukemia Virus Type 1) , FIV (Feline Immunodeficiency Virus) , or FLV (Feline Leukemia Virus) .
  • HIV-1 Human Immunodeficiency Virus Type 1
  • HIV-2 Human Immunodeficiency Virus Type 2
  • HTLV-1 Human T-Cell Leukemia Virus Type 1
  • FIV Feline Immunodeficiency Virus
  • FLV Feine Leukemia Virus
  • the subject in which the above-described composition inhibits replication of a retrovirus is a mammal.
  • the subject may be any mammal.
  • the subject is a human, in which case the composition of the invention particularly inhibits in the subject replication of retroviruses which most commonly infect humans, such as HIV-1 or HIV-2, or HTLV-I.
  • retroviruses which most commonly infect humans such as HIV-1 or HIV-2, or HTLV-I.
  • the composition would still be useful for inhibiting replication of that retrovirus.
  • the subject is a cat, in which case the composition of the invention particularly inhibits replication of retroviruses which most commonly infect cats, including, but not limited to, FIV or FLV.
  • the composition of the invention also inhibits in cats the replication of retroviruses which do not commonly infect cats.
  • composition in the description of above composition is as defined above.
  • Z j is either C-R 5 or N; wherein Z 2 is N; wherein X x and X 2 are the same or different and are 0, S, or N-R 4 ; and wherein R j , R 2 , R 3 , R 4 , and R 5 are the same or different and are either hydrogen or a branched or unbranched alkyl group of from 1 to 3 carbons .
  • administering the pharmaceutical composition to the subject comprises orally administering the composition to the subject.
  • the pharmaceutical composition may be in the form of a capsule, tablet, or solution.
  • the pharmaceutical composition is injected into the subject.
  • Injection may be intramuscular, intraperitoneal, intravenous, or subcutaneous.
  • the pharmaceutical composition may be injected into any part of the subject's body.
  • the pharmaceutical composition is topically applied to the subject. If the pharmaceutical composition is topically applied, the pharmaceutical composition may, for example, be in the form of a lotion or cream.
  • the term "amount effective to inhibit retroviral replication” is intended to mean any amount which will inhibit the replication of the retrovirus in the infected subject. This amount will depend on various factors known to those of ordinary skill in the art. Such factors include, but are not limited to, the size, weight and age of the subject and the concentration of the retrovirus in the infected subject.
  • Retrovirus the replication of which may be inhibited according to the above-described method include, but not limited to HIV- 1, HIV-2, HTLV-1, FIV, and FLV.
  • the subject is a mammal, including, but not limited to, humans and cats.
  • This invention further provides a method of inhibiting replication of a retrovirus in subject infected with the retrovirus which comprises administering to the subject in an amount effective to inhibit retroviral replication the above- described composition which comprises a pharmaceutically acceptable carrier, a first compound which inhibits replication of the retrovirus and a second compound having the structure :
  • Zj is either C-R 5 or N; wherein Z 2 is N; wherein X j and X 2 are the same or different and are O, S, or N-R 4 ; and wherein R j , R 2 , R 3 , R 4 , and R 5 are the same or different and are either hydrogen or a branched or unbranched alkyl group of from 1 to 3 carbons.
  • administering the pharmaceutical composition to the subject comprises orally administering the composition to the subject.
  • the pharmaceutical composition may be in the form of a capsule, tablet, or solution.
  • the pharmaceutical composition is injected into the subject.
  • Injection may be intramuscular, intraperitoneal, intravenous, or subcutaneous.
  • the pharmaceutical composition may be injected into any part of the subject's body.
  • the pharmaceutical composition is topically applied to the subject. If the pharmaceutical composition is topically applied, the pharmaceutical composition may, for example, be in the form of a lotion or cream.
  • Retrovirus the replication of which may be inhibited according to the above-described method include, but not limited to HIV- 1, HIV-2, HTLV-1, FIV, and FLV.
  • the subject is a mammal, including, but not limited to, humans and cats.
  • This invention further provides a pharmaceutical composition for inhibiting tumor-promoter initiated transcription which comprises a pharmaceutically acceptable carrier and a compound having the structure:
  • Z j is either C-R 5 or N; wherein Z 2 is N; wherein X j and X 2 are the same or different and are O, S, or N-R 4 ; and wherein R j , R 2 , R 3 , R 4 , and R 5 are the same or different and are either hydrogen or a branched or unbranched alkyl group of from 1 to 3 carbons.
  • Zj and Z 2 are both N; X j and X 2 are both S; and Rj, R 2 , and R 3 are each methyl, i.e. the compound of the composition is 7-methyl-6, 8- bis (methylthio)pyrrolo [1,2-a]pyrazine, which is sometimes referred to as "metabolite III".
  • the compound of the above composition and therefore the composition itself, is useful for inhibiting transcription which is initiated by a tumor promoter.
  • Tumor promoters are well known to those of ordinary skill in the art and include, but are not limited to, phorbol ester and phenobarbital.
  • the term "tumor promoter” also encompasses complete carcinogens, i.e. compounds which exhibit, inter alia, tumor promoter activity.
  • Complete carcinogens include, but are not limited to, benzpyrene and the polycyclic aromatic hydrocarbons which are found, for example, in cigarette smoke.
  • pharmaceutically acceptable carrier for purposes of the above-described composition is as defined above.
  • administering the pharmaceutical composition to the subject comprises orally administering the composition to the subject. If oral administration is employed, the pharmaceutical composition may be in the form of a capsule, tablet, or solution.
  • the pharmaceutical composition is injected into the subject.
  • Injection may be intramuscular, intraperitoneal, intravenous, or subcutaneous.
  • the pharmaceutical composition may be injected into any part of the subject's body.
  • the pharmaceutical composition is topically applied to the subject. If the pharmaceutical composition is topically applied, the pharmaceutical composition may, for example, be in the form of a lotion or cream.
  • the term "amount effective to inhibit tumor formation” is intended to mean any amount which will inhibit the formation of tumors in the subject. This amount will depend on various factors known to those of ordinary skill in the art. Such factors include, but are not limited to, the size, weight and age of the subject.
  • the subject is a mammal. In one embodiment of the above-described method wherein the subject is a mammal, the subject is a human.
  • the subject is likely to be exposed to a tumor promoter, such as any of the tumor promoters, including the complete carcinogens, identified above.
  • a tumor promoter such as any of the tumor promoters, including the complete carcinogens, identified above.
  • a subject who is likely to be exposed to cigarette smoke is a subject likely to be exposed to tumor promoters.
  • the subject is predisposed to developing tumors.
  • Subjects predisposed to developing tumors are well known to those of ordinary skill in the art.
  • An example of a subject predisposed to developing tumors is a subject predisposed to having cancer, such as a subject with ulcerative colitis, which subject is therefore predisposed to developing colon cancer.
  • This invention further describes a compound having the structure:
  • Z x is either C-R 5 or N; wherein Z 2 is N; wherein X, and X 2 are the same or different and are O, S, or N-R 4 ; and wherein R j , R 2 , R 3 , R 4 , and R 5 are the same or different and are either hydrogen or a branched or unbranched alkyl group of from 1 to 3 carbons; with the proviso that when Zj and Z 2 are both N, X j and X 2 are both S, and Rj and R 2 are both methyl, R 3 is not methyl.
  • the above-described compounds are useful for inhibiting the replication of retroviruses and for inhibiting tumor promoter initiated transcription.
  • HIV-1 human immunodeficiency virus type 1
  • RT reverse transcriptase
  • PBMC peripheral blood mononuciear cell
  • PBS phosphate-buffered saline
  • PHA-P phytohemagglutinin P
  • IL-2 interleukin-2
  • DMSO dimethylsulfoxide
  • MTT 3- (4, 5-dimethylthiazo-2-yl) -2, 5- diphenyltetrazolium bromide
  • PMA phorbol-12-myristate-13- acetate
  • EGTA ethylene glycol bis (/3-aminoethyle ether) - N,N,N' ,N' -tetraacetic acid
  • NAC n-acetyl-L-cysteine.
  • PBMCs were obtained from the New York Blood Center as buffy coats; cell lines were obtained from the National Institutes of Health AIDS Research and Reference Reagent Program
  • Metabolite III was prepared according to the method of Fleury et al. (13) . Briefly, oltipraz (O.llg, 0.50mmol) was dissolved 500ml of absolute ethanol under argon and heated to 35 ° , yielding a deep orange- colored solution. While this mixture was being stirred, 20ml of a yellow-brown solution of absolute ethanol containing 5mmol of sodium ethoxide and 0.30g (2.5mmol) of L-cysteine were added via hypodermic syringe over 5 minutes.
  • oltipraz O.llg, 0.50mmol
  • Cell lines were grown in RPMI 1640 medium supplemented with 20% fetal bovine serum, 2 mM glutamine, 100 units/ml penicillin, and lOO ⁇ g/ml streptomycin, at 37° in humidified incubators with 5% C0 2 .
  • PBMCs isolated from buffy coats with a ficoll density gradient, were washed three times in PBS and were grown in the aforementioned medium supplemented with l ⁇ g/ml PHA-P. After 24 hour exposure to PHA-P, the supernatant was replaced with medium supplemented with 5% IL-2 (without PHA-P) .
  • Oltipraz and metabolite III were dissolved in DMSO and were stored at -70 ° , The stabilities of oltipraz and metabolite III in DMSO were assessed by high performance liquid chromatography using a Microsorb C 18 column (Ranin, Emeryville, CA) run under isocratic elution conditions (methanol/water, 80:20) , and the antiretroviral activities of old stocks were routinely compared with those of freshly prepared stocks at the time that the old stocks were replaced. There was neither chemical nor biological evidence for degradation of either drug in DMSO stored at -70 ° for >1 year (data not shown) . The DMSO stocks were diluted into medium, to 0.2% maximal final concentration, just before treatment of the cells. Because the cells were cultured with 20% fetal bovine serum, no obvious precipitation of oltipraz or metabolite III was observed with final concentrations up to 200 ⁇ M.
  • H9 and CEM cells were infected with HIV-1 (isolate HTLV-IIIB) at 1000 50% tissue culture infectious doses/10 6 cells. Virus was allowed to adsorb for 1 hour, after which the unadsorbed virus was removed by centrifugation. After the cells were washed twice with PBS, they were added to 96-well microtiter plates with medium containing up to 0.2% DMSO and various concentrations of oltipraz or metabolite III, at a final density of 500,000 cells/ml (final volume/well, 200 ⁇ l) .
  • HIV-1 isolated HTLV-IIIB
  • Virus was allowed to adsorb for 1 hour, after which the unadsorbed virus was removed by centrifugation. After the cells were washed twice with PBS, they were added to 96-well microtiter plates with medium containing up to 0.2% DMSO and various concentrations of oltipraz or metabolite III, at a final density of 500,000 cells/ml
  • Ul and ACH-2 cells were treated as described (10) , by incubating the cells (50,000 cells/microtiter well, 200- ⁇ l final volume) with various concentrations of oltipraz or metabolite III for 24 hours before the addition of 2.5 nM PMA. The cells were grown for an additional 48 hours, and p24 antigen release was measured in the culture supernatants.
  • PBMCs were used for experiments 2-6 days after PHA-P stimulation. Except for two experiments, PBMCs for each experiment were obtained from different donors .
  • the PBMCs were treated as described for H9/CEM cells lines, except that the cells were plated at a density of 10 6 cells/ml in medium supplemented with 5% IL-2.
  • Cytotoxicity assays Uninfected PBMCs, H9 cells, and CEM cells were treated as described above for infected cells . On day 7, cell densities were estimated by MTT staining (10) . For Ul and ACH-2 cells, the cytotoxicity of oltipraz and metabolite III was assessed by MTT staining of matched non-
  • Isotopic RT assays were performed as described (16) , except that 4 ⁇ M (final concentration) unlabeled dTTP was added to all incubation mixtures. Assays were performed in duplicate or triplicate.
  • the U293.27.2 cell line is a stably-transfected clone derived from the 293S embryonic human kidney cell line which contains a construct of HIV-1 LTR fused to the bacterial ⁇ - galactosidase gene (32) .
  • Cells were plated at a density of 3000 cells per 0.32 cm 2 well as described (32) in triplicate set of plates utilizing the plate format outlined by Prochaska (33) . Cells were pretreated with test compounds 24 hours after plating and 0-48 hours prior to the addition of 10 ng/ml PMA (upper four rows of each plate) .
  • One set of plates were assayed for 3-galactosidase as described (32) 24 hours after the addition of PMA except that the formation of 0-nitrophenol from 0-nitropheyl-3-D-galactopyranoside was determined spectrophotometrically with a microtiter plate scanner at 405nm. Cell densities were determined by staining the remaining sets of plates with either crystal violet (34) or MTT (35) .
  • ⁇ -galactosidase specific activities were determined by normalizing the jS-galactosidase activity to the MTT [3- (4,5- dimethylthiazo-2-yl) -2, 5-diphenyltetrazolium bromide] staining for all compounds tested except NAC, where ⁇ -galactosidase activities were normalized to crystal violet staining.
  • the treated-to-control ratios were determined by normalizing the drug-treated specific activities to the specific activities of the PMA-stimulated, vehicle (DMSO) -treated control cells on the same plate.
  • DMSO PMA-stimulated, vehicle
  • Metabolite III was synthesized in 91% yield according to the method of Fleury et al. (13) .
  • the synthesis of metabolite III from cysteine demonstrates that oltipraz is an electrophile.
  • oltipraz may be relevant to its biological properties, because oltipraz is thought to inactivate and/or modify proteins that are critical to its antischistosomal (i.e. schistosomal glutathione S- transferases) (18) , anticarcinogenic (i.e., transcription factors) (19,20), and antiviral (i.e., RT) (9) activities.
  • antischistosomal i.e. schistosomal glutathione S- transferases
  • anticarcinogenic i.e., transcription factors
  • RT antiviral
  • Metabolite III was tested as an antiviral agent because it is the major metabolic intermediate of oltipraz in vivo and the results with PMA-stimulated Ul cells suggested that oltipraz may be a prodrug in this chronic infection model (10) .
  • metabolite III treatment resulted in a concentration-dependent inhibition of p24 antigen release into culture supernatant (Fig.2) .
  • the ED 50 was approximately 25 ⁇ M.
  • Oltipraz was approximately 2.5-fold more potent than metabolite III.
  • Oltipraz and metabolite III inhibited viral replication in H9 cells with similar potencies, using indirect immunofluorescence as a surrogate marker for viral replication (data not shown) .
  • the assays examining the antiviral activity of metabolite III and oltipraz were not performed above 60 ⁇ M, cytotoxicity studies in uninfected cells indicated that metabolite III was relatively nontoxic (Table 1) .
  • the selectivity index (IC 50 /ED 5 o) for metabolite III was 6.6 in H9 cells and 4.4 in CEM cells.
  • oltipraz acts as an electrophile by covalently modifying a nucleophilic amino acid residue of HIV- 1 RT.
  • metabolite III had negligible effects on purified recombinant HIV-1 RT when assayed for its ability to reversibly or irreversibly inhibit the enzyme (Fig. 3) .
  • the inability of metabolite II to inactivate RT is not surprising, because metabolite III is not an electrophile.
  • ACH-2 cells were particularly useful for narrowing the potential mechanisms by which oltipraz and metabolite III inhibit HIV-1 replication.
  • oltipraz was active as an inhibitor of HIV-1 replication in acutely infected H9 and CEM T cell lymphoma models (Fig.2)
  • oltipraz was devoid of antiviral activity in chronically infected ACH-2
  • T cell lymphoma T cell lymphoma cells (Fig.4) .
  • differences in the uptake and metabolism of oltipraz are responsible for the inactivity of oltipraz in ACH-2 cells, that is less likely than an intrinsic difference in the mechanism of action of oltipraz and metabolite III, because ACH-2 cells are ultimately derived from the CEM cell line.
  • the ability to inhibit acute, but not chronic, replication in these T cell models supports our proposal that oltipraz inhibits HIV-1 replication via the inactivation of RT (9) .
  • metabolite III is an effective inhibitor of HIV-1 replication in ACH-2 cells, suggesting that metabolite III acts to inhibit viral replication at a step distal to the integration of the viral genome.
  • metabolite III could inhibit HIV-1 replication in ACH- 2 cells
  • metabolite III was tested for the ability to inhibit HIV-1 transcription.
  • metabolite III could inhibit the LTR-driven transcription of /3-galactosidase in stably transfected 293S human embryonic kidney cells (clone 293.27.2) (31) .
  • Cells were pretreated with test compounds 0-48 h prior to the addition 10 ng/ml PMA 72 hours after plating the cells, and were assayed for ⁇ - galactosidase 24 hours after PMA stimulation.
  • NAC N-acetyl- L-cysteine was included as a positive control, since it is able to inhibit HIV-1 replication and LTR-driven transcription (31, 32) .
  • FIG. 1 shows that metabolite III is able to inhibit the LTR-driven transcription of HIV-1 metabolite III in PMA- stimulated cells with an IC 50 15 ⁇ M, in good agreement with IC 50 values determined for HIV-1 replication in a number of T- cell models.
  • the effects of metabolite III were essentially identical to the results obtained with NAC, except that metabolite III is approximately 1000-fold more potent.
  • ED 50 for metabolite III was 18 ⁇ M (eight independent experiments) .
  • the dose-response curve for inhibition of viral replication by metabolite III was much shallower than that for oltipraz. It was also clear that the inhibitory activity of metabolite III varied more markedly than did that of oltipraz. For example, in three of eight independent experiments the ED 50 for metabolite III was ⁇ 5 ⁇ M. Whether this reflects differences in donor lymphocytes and/or the time at which stimulated PBMCs were exposed to metabolite
  • cytotoxicities of oltipraz and metabolite III in PBMCs are given in Table 1.
  • the selectivity indices for oltipraz and metabolite III in PBMCs were 5- and 3- fold, respectively.
  • H9 cells CEM cells PBMCs ⁇ M
  • metabolite III is formed from oltipraz in vivo, determination of whether they were synergistic was sought.
  • the dose-response curves for oltipraz, metabolite III, and oltipraz/metabolite III (2:1) are shown in Fig. 5, and the combination indices calculated from these data are shown in Fig. 5, inset.
  • Oltipraz and metabolite III were synergistic at fractional inhibitions of >15% (a combination index of ⁇ 1 indicates synergy) .
  • oltipraz and metabolite III are synergistic inhibitors of HIV-1 replication raises questions regarding the potential in vivo activity of oltipraz, especially because most of the absorbed drug undergoes rearrangement to metabolite III (11,12) .
  • rearranged metabolites related to metabolite III represent >75% of 14 C-labeled oltipraz recovered from the serum or urine (11,12) .
  • the serum concentration of oltipraz at nontoxic doses in humans and rodents has been documented to be in the low micromolar range (up to 20 ⁇ M) (3-8) .
  • oltipraz and its arranged metabolites are found in serum at concentrations that inhibit HIV-1 replication in vi tro, and the above-described results suggest that oltipraz and its metabolites could act to synergistically inhibit HIV-1 replication in vivo .
  • metabolite III inhibits HIV-1 replication in vi tro. This is the first reported biological activity for this compound.
  • metabolite III appears to be an inhibitor of HIV-1 replication at a point in the replicative life cycle that is distal to viral integration. Therefore, just as oltipraz represents a new lead for the design of inhibitors of acute replication (and RT) , metabolite III represents a new lead compound for the prevention of HIV-1 replication in chronically infected cells.
  • Oltipraz and metabolite III inhibit HIV-1 replication via different mechanisms, and the results with PBMCs demonstrate that oltipraz and metabolite III are synergistic.
  • oltipraz may be a more effective antiretroviral agent in vivo than it is in vi tro .
  • This possibility suggests a clinical trial evaluating the ability of oltipraz to reduce viral load which should include an examination of oltipraz and metabolite III pharmacokinetics.
  • many of the known biological effects of oltipraz have been attributed to the parent compound (9, 18, 20)
  • the above results demonstrate that oltipraz is a pro-drug, albeit affecting very different biological processes. It is not surprising that the biological properties of oltipraz and metabolite III are so distinct since the structures of the two compounds differ significantly.
  • oltipraz antischistosomal, anticarcinogenic, and antiviral activities of the parent compound (oltipraz) are thought to be mediated by the ability of the drug to covantly modify target proteins (i.e. schistosomal glutathione S-transferase (18) , transcription factors involved in the regulation of Phase II enzymes (20) , and RT (9) , respectively) .
  • target proteins i.e. schistosomal glutathione S-transferase (18) , transcription factors involved in the regulation of Phase II enzymes (20) , and RT (9) , respectively.
  • metabolite III a) is not an electrophile, b) does not induce Phase II enzymes nor activate EpRE (Electrophile Response Element) -driven transcription) (20) ; and c) does not inhibit nor inactivate HIV-1 RT.
  • metabolite III may represent a lead compound for the identification of nontoxic inhibitors of mitogenic signal transduction.
  • oltipraz may have greater antiretroviral activity in vivo than in vitro, and the potential effect of metabolite III as an antagonist of tumor promoter signal transduction may account for the considerable chemopreventive activity that oltipraz is known to possess.
  • oltipraz an inhibitor of human immunodeficiency virus type 1 replication in vi tro (IC 50 - lO ⁇ M) , undergoes extensive metabolism in vivo .
  • Most of the orally administered drug undergoes opening of the dithiolethione ring, reduction, recyclization, and methylation to form 7-methyl-6, 8- bis (methylthio)pyrrolo [1, 2-a]pyrazine ("metabolite III”) .
  • the phar acokinetics of oltipraz and metabolite III can be compared with the pharmacodynamic effects of orally administered oltipraz in a phase I/II trial of oltipraz in patients with p24 antigenemia.
  • HIV-1 human immunodeficiency virus type 1
  • Metabolite III but not oltipraz, inhibit phorbol 12- myristate 13-acetate (PMA) -stimulated viral replication in • chronically infected ACH-2 T-cell lymphoma cells, suggesting that metabolite III disrupt viral replication at a point in the HIV-1 life-cycle that is distal to integration of the viral genome.
  • PMA phorbol 12- myristate 13-acetate
  • metabolite III but not oltipraz, inhibits long-terminal repeat (LTR) -driven expression of ⁇ - galactosidase in PMA-stimulated cells (IC 50 ⁇ 15 MM) .
  • LTR long-terminal repeat
  • ⁇ - galactosidase ⁇ - galactosidase
  • RT reverse transcriptase
  • oltipraz prevents the initiation phase of carcinogenesis by diverting ultimate carcinogens from DNA (2, 22, 23), whereas the rearranged metabolite may block the growth and selection of initiated cells by mitogens and cytokines.
  • the known antipromoting effects of oltipraz (24) may be due to the effects of rearranged metabolite, and may rationalize why oltipraz is a more effective and complete chemopreventive agent than can be appreciated by solely considering the capacity of oltipraz to act as an inducer Phase II enzymes.
  • Tumor necrosis factor ⁇ induces expression of human immunodeficiency virus in a chronically infected T-cell clone. Proc. Natl . Acad. Sci . USA 86:2365-2368 (1989) .
  • Kensler, T. W. Egner, P. A., Dolan, P. M. , Groopman, J. D. & Roebuck, B. D. Cancer Res . 47, 4271-4277 (1987) .

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Abstract

La présente invention concerne une composition pharmaceutique pour inhiber la réplication d'un rétrovirus, contenant un composé ayant la structure (I). Dans cette formule, Z1 est C-R5 ou N; Z2 est N; X1 et X2 sont les mêmes ou différents et représentent O, S ou N-R4; et R1, R2, R3, R4 et R5 sont les mêmes ou différents et représentent soit hydrogène, soit un groupe alkyle ramifié ou droit ayant 1 à 3 atomes de carbone. L'invention concerne, en outre, une composition pharmaceutique pour inhiber la réplication d'un rétrovirus qui comprend un premier composé qui inhibe la réplication du rétrovirus, et un second composé ayant la structure définie précédemment; des procédés pour inhiber la réplication rétrovirale chez un sujet, à l'aide des compositions selon l'invention; une composition pharmaceutique pour inhiber une transcription induite par un promoteur de tumeur, qui comprend un composé ayant la structure définie ci-dessus; et un procédé pour empêcher la formation de tumeurs chez un sujet, consistant à administrer ladite composition pour inhiber une transcription induite par un promoteur de tumeur.
PCT/US1996/011699 1995-07-13 1996-07-12 Composes, compositions et procedes pour inhiber la replication de retrovirus et pour inhiber une transcription induite par un promoteur de tumeur WO1997003055A1 (fr)

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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999060988A2 (fr) * 1998-05-29 1999-12-02 University Of Florida Therapie combinee pour le traitement des infections dues au vif
US6875773B1 (en) 1998-05-29 2005-04-05 Ben M. Dunn Combination therapy for treatment of FIV infection
US7199122B2 (en) 2001-10-02 2007-04-03 Fox Chase Cancer Center Methods for inhibiting angiogenesis
CN102933584A (zh) * 2010-04-16 2013-02-13 Abbvie公司 激酶的吡咯并吡嗪酮抑制剂
US20150031701A1 (en) * 2007-03-12 2015-01-29 St Ip Holding Ag Compositions and methods for preventing and treating mucositis and weight loss
US11135220B1 (en) 2020-04-08 2021-10-05 St Ip Holding Ag Methods of treating viral infections with formulated compositions comprising 4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiole-3-thione

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
MOLECULAR PHARMACOLOGY, July 1995, Vol. 48, No. 1, PROCHASKA et al., "Inhibition of Human Immunodeficiency Virus Type 1 Replication by 7-Methyl-6,8-bis(Methylthio)pyrrolo(1,2-a) Pyrazine, an In Vivo Metabolite of Oltipraz", pages 15-20. *
TETRAHEDRON LETTERS, 1982, Vol. 23, No. 35, CORBET et al., "A Novel Cyclization Reaction Leading to Pyrrolo (1,2-a) Pyrazines", pages 3565-3566. *
TETRAHEDRON, 1985, Vol. 41, No. 18, FLEURY et al., "Studies of the Reaction of 1,2-Dithiole-3-thiones with Nucleophiles", pages 3705-3715. *
TETRAHEDRON, 1987, Vol. 43, No. 15, LARGERON et al., "Reactivity of Substituted 1,2-Dithiole-3-thiones with Sodium Ethanethiolate: A Convenient Route to a Novel Heterocycle", pages 3421-3428. *

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999060988A2 (fr) * 1998-05-29 1999-12-02 University Of Florida Therapie combinee pour le traitement des infections dues au vif
WO1999060988A3 (fr) * 1998-05-29 2000-12-07 Univ Florida Therapie combinee pour le traitement des infections dues au vif
US6875773B1 (en) 1998-05-29 2005-04-05 Ben M. Dunn Combination therapy for treatment of FIV infection
US7199122B2 (en) 2001-10-02 2007-04-03 Fox Chase Cancer Center Methods for inhibiting angiogenesis
US7452884B2 (en) 2001-10-02 2008-11-18 Fox Chase Cancer Center Methods for inhibiting angiogenesis
US20150031701A1 (en) * 2007-03-12 2015-01-29 St Ip Holding Ag Compositions and methods for preventing and treating mucositis and weight loss
CN102933584A (zh) * 2010-04-16 2013-02-13 Abbvie公司 激酶的吡咯并吡嗪酮抑制剂
US11135220B1 (en) 2020-04-08 2021-10-05 St Ip Holding Ag Methods of treating viral infections with formulated compositions comprising 4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiole-3-thione
WO2021205233A3 (fr) * 2020-04-08 2021-12-16 St Ip Holding Ag Procédés de traitement d'infections virales avec des compositions formulées comprenant de la 4-méthyl-5-(pyrazin-2-yl)-3h-1,2-dithiole-3-thione

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