CK-II Abstracts
History Quercetin Spirulina Baicalein Quercetin and Vitamin C CK-II Inhibitors CK-II Abstracts Winning Game Dosage Strategies
I published this article on my Yahoo site "Nat_anti_hiv" in association with the article "CK-II Inhibitors"
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I have provided copies of the abstracts here for your convenience. Refer to
actual articles for more details.
This article can be read in association with the article
"CK2 Inhibitors", for my interpretation of research and the
implications for treatment of HIV infection.
All abstracts are from Medline:
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The following is an abstract from an important article describing the action of Rev.
My summary:
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[1]
FEBS Lett 1998 May 29;428(3):235-40
Biochemical characterization of HIV-1 Rev as a
potent activator of casein kinase II in vitro.
Ohtsuki K, Maekawa T, Harada S, Karino A, Morikawa Y,Ito M
Laboratory of Genetical Biochemistry, Kitasato University School of Allied Health Sciences, Sagamihara, Japan. [email protected]
The stimulatory effects of several DNA-binding basic proteins (histone and protamine) and HIV-1 Rev with arginine (Arg)-rich clusters on the activity of casein kinase II (CK-II) were investigated in vitro. It was found that recombinant Rev (rRev) and the synthetic oligo-fragments corresponding to the amino acid sequences of its Arg-rich cluster stimulate CK-II activity in a dose-dependent manner. The activated CK-II phosphorylates several cellular and viral proteins in HIV-1 infected human MOLT-4 cells, and also phosphorylates HIV-1 structural proteins, including recombinant reverse transcriptase (rRT). These phosphorylations are selectively
inhibited by CK-II inhibitors, such as quercetin, oGA (a glycyrrhetinic acid derivative) and NCS-chrom (an enediyne containing antibiotic). The data presented here
suggest that HIV-1 Rev acts as an effective potent activator of CK-II, which may be a cellular mediator promoting HIV-1 replication in virus-infected cells.
PMID: 9654140, UI: 98316712
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This abstract focuses on CK2 phosphorylation of the viral enzyme Reverse Transcriptase.
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[2]
Biol Pharm Bull 1999 Oct;22(10):1122-6
Casein kinase II (CK-II)-mediated stimulation of HIV-1 reverse transcriptase activity and characterization of selective inhibitors in vitro.
Harada S, Haneda E, Maekawa T, Morikawa Y, Funayama S, Nagata N, Ohtsuki K, Nagata N,
Ohtsuki K
Laboratory of Genetical Biochemistry, School of Allied Health Sciences, Kitasato University, Sagamihara, Japan.
The physiological significance of the casein kinase II (CK-II)-mediated phosphorylation of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) on its three enzymatic activities [RNA-dependent DNA polymerase (RDDP), DNA-dependent DNA polymerase (DDDP) and ribonuclease H (RNase H)] was investigated in vitro. It was found that (i) the purified recombinant RT (rRT) functioned as an effective phosphate acceptor for CK-II; (ii) the RDDP, DDDP and RNase H activity of rRT was stimulated about 2.8-, 4.1- and 3.9-fold, respectively, after full phosphorylation by CK-II; and (iii) this stimulation was selectively inhibited by potent CK-II inhibitors, such as neocarzinostatin-chromophore (NCS-chrom) and three polyphenol-containing anti-oxidant compounds
[quercetin, epigallocatechin gallate (EGCG) and 8-chloro-3',4',5,7-tetrahydroxyisoflavone (8C-3',4',5,7-THI)]. These results suggest that (i) CK-II may be responsible for activation of RT in HIV-1-infected cells; and (ii) the selective inhibition of CK-II-mediated activation of HIV-1 RT by potent CK-II inhibitors may be involved in the mechanism of their anti-HIV-1 effects at the cellular level.
PMID: 10549869, UI: 20016062
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This is an earlier article, that showed an earlier recognition of the role of CK-II in viral production
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[3]
Proc Natl Acad Sci U S A 1997 Jun 10;94(12):6110-5
Casein kinase II is a selective target of HIV-1 transcriptional inhibitors.
Critchfield JW, Coligan JE, Folks TM, Butera ST
Retrovirus Diseases Branch, Division of Acquired Immunodeficiency Syndrome, Sexually Transmitted Diseases, and Tuberculosis Laboratory Research, Centers for Disease Control and Prevention, Atlanta, GA 30333, USA. [email protected]
The identification of cellular factors that are required to complete various steps of the HIV-1 life cycle may lead to the development of new therapeutics. One key step, transcription from the integrated provirus, is inhibited by members of two distinct classes of compounds, the flavonoids and the benzothiophenes, via an unknown mechanism, possibly involving a cellular factor. A marked specificity toward inhibiting HIV-1 transcription is evidenced by the ability of drug-treated cells to retain their proliferative and differentiation capabilities. In addition, the compounds do not impede the activation and function of the transcriptional factor NF-kappaB. Here we report on the identification of several cellular proteins that mediate the HIV-1 transcriptional inhibitory property of the flavonoid chrysin.
Chemical and immunologic analyses identified these cellular proteins as the individual subunits of casein kinase II (CKII). Though structurally unrelated to chrysin, an HIV-1 inhibitory benzothiophene also bound selectively to CKII. Both chrysin and the benzothiophenes inhibited human recombinant CKII enzymatic activity and showed competitive kinetics with respect to ATP, analogous to the classic CKII inhibitor 5, 6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB). Moreover,
DRB potently inhibited HIV-1 expression in chronically infected cells. CKII may regulate HIV-1 transcription by phosphorylating cellular proteins involved in HIV-1 transactivation that contain multiple CKII phosphorylation consensus sequences.
PMID: 9177178, UI: 97322333
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The following abstract indicates that Quercetin among others inhibits the viral enzyme protease. It seems to me that other work indicates that the mechanism of inhibition is once again inhibition of phosphorylation of viral enzymes by activated CK-II.
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[4]
Biol Pharm Bull 2000 Sep;23(9):1072-6
Inhibitory activity of flavonoids and tannins against HIV-1 protease
Xu HX, Wan M, Dong H, But PP, Foo LY
Department of Biology and Institute of Chinese Medicine, The Chinese University of Hong Kong.
[Record supplied by publisher]
Twenty-nine flavonoids and six hydrolyzable tannins were studied for their inhibitory activity against human immunodeficiency virus (HIV)-1 protease using fluorescence and HPLC assays. Among the flavonoids, flavones, flavanones, flavonols, catechols and chalcones, the flavonols were the most active category while flavanones and catechols displayed low activity. Quercetin was the most potent inhibitor of the target enzyme with an IC50 value of 58.8 microM, while butein and luteolin showed moderate activity. Of the hydrolyzable tannins tested, three ellagitannins which contain a hexahydroxvdiphenoyl (HHDP) unit linked to the O-3 and 0-6 positions of the sugar, were found to strongly inhibit HIV-1 protease. The IC50 values of corilagin and repandusinic acid on HIV-1 protease were 20.7 and 12.5 microM, respectively.
PMID: 10993207
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The following abstract is an earlier one indicating certain substances, (e.g. Quercetin) inhibit the viral enzyme Integrase. I believe later work supports the hypothesis that the inhibition is due to inhibition of activated CK-II, which explains why DNA binding was not correlated with Integrase inhibition
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[5]
Proc Natl Acad Sci U S A 1993 Mar 15;90(6):2399-403
Inhibitors of human immunodeficiency virus integrase.
Fesen MR, Kohn KW, Leteurtre F, Pommier Y
Laboratory of Molecular Pharmacology, National Cancer Institute, Bethesda, MD 20892.
In an effort to further extend the number of targets for development of antiretroviral agents, we have used an in vitro integrase assay to investigate a variety of chemicals, including topoisomerase inhibitors, antimalarial agents, DNA binders, naphthoquinones, the flavone quercetin, and caffeic acid phenethyl ester as potential human immunodeficiency virus type 1 integrase inhibitors. Our results show that although several topoisomerase inhibitors--including doxorubicin, mitoxantrone, ellipticines, and quercetin--are potent integrase inhibitors, other topoisomerase inhibitors--such as amsacrine, etoposide, teniposide, and camptothecin--are inactive. Other intercalators, such as chloroquine and the bifunctional intercalator ditercalinium, are also active. However, DNA binding does not correlate closely with integrase inhibition. The intercalator 9-aminoacridine and the polyamine DNA minor-groove binders spermine, spermidine, and distamycin have no effect, whereas the non-DNA binders primaquine, 5,8-dihydroxy-1,4-naphthoquinone, and caffeic acid phenethyl ester inhibit the integrase. Caffeic acid phenethyl ester was the only compound that inhibited the integration step to a substantially greater degree than the initial cleavage step of the enzyme.
A model of 5,8-dihydroxy-1,4-naphthoquinone interaction with the zinc finger region of the retroviral integrase protein is proposed.
PMID: 8460151, UI: 93211969
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This article reports that Quercetin which is a known CK2 inhibitor inhibits Vpr-induced cell cycle perturbation. This implies that phosphorylation of Vpr is inhibited.
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[6]
Biochem Biophys Res Commun 1999 Aug 2;261(2):308-16
Inhibition of Vpr-induced cell cycle abnormality by quercetin: a novel strategy for searching
compounds targeting Vpr.
Shimura M, Zhou Y, Asada Y, Yoshikawa T, Hatake K, Takaku F, Ishizaka Y
Department of Intractable Diseases, International Medical Center of Japan, 1-21-1 Toyama, Shinjuku-ku, Tokyo, 162-8655, Japan.
Vpr, an accessory gene product of HIV-1 which induces cell cycle abnormality leading to the increased HIV replication, is supposed to be a possible target for
anti-AIDS drugs. We recently established a cell line (MIT-23) in which Vpr-induced cell cycle perturbation could be manipulated by a tetracycline promoter.
Here, we screened anti-Vpr activity in 27 kinds of herb drugs using MIT-23 cells. One of the extracts prepared from Houttuyniae herba showed an inhibitory
activity. Quercetin (QCT), a compound of this crude drug, efficiently inhibited Vpr function without affecting its expression. Furthermore, data suggested that Vpr-induced transcription from HIV-LTR was considerably abrogated by QCT. These data indicate that QCT, a flavonoid previously reported to inhibit HIV replication, also targets Vpr, implicating that MIT-23 cell provides a novel strategy for screening compounds possessing anti-Vpr activity which would be in turn utilized for clarifying the mechanism of Vpr function.
PMID: 10425183, UI: 99355569
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The following abstract suggests to me that although up-regulation of the virus is inhibited by CK-II inhibitors, low-level replication continues. A possible hypothesis from reading the other articles, for why the highly replicating cells have disappeared, is that they are converted to lower viral production by inhibition of activated CK-II up-regulation. They haven't been destroyed. This also explains why there are more cells with lower level replication. In this case the CK-II inhibitor is Baicalein. This implies that treatment should also include direct inhibitors of viral enzymes, to reduce low-level replication - for instance current combination therapy.
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[7]
Jpn J Med Sci Biol 1995 Apr;48(2):79-87
Apoptosis of HIV-infected cells following treatment with Sho-Saiko-to and its components.
Wu X, Akatsu H, Okada H
Department of Molecular Biology, Nagoya City University School of Medicine.
Baicalein and baicalin are components of Sho-saiko-to (SST), a Chinese medical drug which is claimed to be therapeutically effective in treating HIV-infected patients. Although 20 micrograms/ml of baicalin was not cytotoxic to CEM cells, a cultured T cell line, it proved to be cytotoxic to HIV-infected CEM cells (CEM-HIV) with a higher HIV-releasing capacity and DNA fragmentation was detected within 24 hr of incubation. However, after incubation of CEM-HIV with a lower dose of baicalin (0.1, 0.3 and 2 micrograms/ml) for 24 and 48 hr, the viable cell number increased by about 25% and the p24 release into the medium was 25% lower than that of the control. After further incubation in the presence of the agent for 6 and 9 days, only cells with a lower HIV-releasing capacity survived. Baicalin might selectively induce apoptosis of CEM-HIV cells which have a high virus-releasing capacity, and stimulate proliferation of CEM-HIV which have a relatively lower capacity of HIV-production.
PMID: 7474502, UI: 96022735
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The
following abstract suggests to me the importance of using ascorbate (vitamin C)
with Quercetin
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[8]
Antiviral activity of flavones and potentiation by ascorbate
R Vrijsen, L Everaert and A Boeye
Department of Microbiology and Hygiene, Vrije Universiteit
Brussel, Belgium.
We compared the anti-poliovirus activities of three flavones, quercetin, luteolin and 3-methylquercetin, which differ only at ring position 3. 3-Methylquercetin was the most potent compound. Quercetin exhibited antiviral activity only when protected against oxidative degradation by ascorbate. The antiviral activity of luteolin was comparable to that of ascorbate-stabilized quercetin.
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This is an article about Spirulina. Note comment about syncytium formation and the necessity of chelation with calcium.
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[9]
AIDS Res Hum Retroviruses 1996 Oct 10;12(15):1463-71
A natural sulfated polysaccharide, calcium spirulan, isolated from Spirulina platensis: in vitro
and ex vivo evaluation of anti-herpes simplex virus and anti-human immunodeficiency virus
activities.
Hayashi K, Hayashi T, Kojima I
Department of Virology, Toyama Medical and Pharmaceutical University, Japan.
A sulfated polysaccharide named calcium spirulan (Ca-SP) has been isolated from a sea alga, Spirulina platensis, as an antiviral component. The anti-human
immunodeficiency virus type 1 (HIV-1) and anti-herpes simplex virus type 1 (HSV-1) activities of Ca-SP were compared with those of dextran sulfate (DS) as a representative sulfated polysaccharide. Anti-HIV-1 activities of these agents were measured by three different assays: viability of acutely infected CD4-positive cells, or a cytopathology assay; determination of HIV-1 p24 antigen released into culture supernatants; and inhibition of HIV-induced syncytium formation.
Anti-HSV-1 activity was assessed by plaque yield reduction. In addition, their effects on the blood coagulation processes and stability in the blood were
evaluated. These data indicate that Ca-SP is a potent antiviral agent against both HIV-1 and HSV-1. Furthermore, Ca-SP is quite promising as an anti-HIV agent because even at low concentrations of Ca-SP an enhancement of virus-induced syncytium formation was not observed, as was observed in DS-treated cultures, Ca-SP had very low anticoagulant activity, and showed a much longer half-life in the blood of mice when compared with that of DS. Thus, Ca-SP can be a candidate agent for an anti-HIV therapeutic drug that might overcome the disadvantages observed in many sulfated polysaccharides. When the role of chelation of calcium ion with sulfate groups was examined by removing calcium or its replacement by sodium, the presence of calcium ion in the molecule was shown to be essential for the dose-dependent inhibition of cytopathic effect and syncytium formation induced by HIV-1.
PMID: 8893054, UI: 97048218
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The following abstract gives indication that CK-II inhibition will give Nervous system side effects. Note that there are substrates for CK-II in synaptic compartments. This may account for anti-depressant effect. The article also indicates that CK-II is unlikely to mutate, since it is "highly conserved in evolution".
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[10]
Prog Neurobiol 2000 Feb;60(3):211-46
Casein kinase 2 as a potentially important enzyme in the nervous system.
Blanquet PR
Unite de Recherche de Physiopharmacologie du Systeme Nerveux, U-161 INSERM, Paris, France. [email protected]
Protein kinase CK2 is a ubiquitous and pleiotropic seryl/threonyl protein kinase which is highly conserved in evolution indicating a vital cellular role for this kinase.
The holoenzyme is generally composed of two catalytic (alpha and/or alpha') and two regulatory (beta) subunits, but the free alpha/alpha' subunits are catalytically active by themselves and can be present in cells under some circumstances. Special attention has been devoted to phosphorylation status and structure of these enzymic molecules, however, their regulation and roles remain intriguing. Until recently, CK2 was believed to represent a kinase especially required for cell cycle progression in non-neural cells. At present, with respect to recent findings, four essential features suggest potentially important roles for this enzyme in specific neural functions: (1) CK2 is much more abundant in brain than in any other tissue; (2) there appear to be a myriad of substrates for CK2 in both synaptic and nuclear
compartments that have clear implications in development, neuritogenesis, synaptic transmission, synaptic plasticity, information storage and survival; (3) CK2 seems to be associated with mechanisms underlying long-term potentiation in hippocampus; and (4) neurotrophins stimulate activity of CK2 in hippocampus. In addition, some data are suggestive that CK2 might play a role in processes underlying progressive disorders due to Alzheimer's disease, ischemia, chronic alcohol exposure or immunodeficiency virus HIV. The present review focuses mainly on the latest data concerning the regulatory mechanisms and the possible neurophysiological functions of this enzyme.
Publication Types:
Review
Review, academic
PMID: 10658642, UI: 20121729
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This article supports the hypothesis that CK2 phosphorylation of viral enzymes is involved in the development of drug resistance
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[11]
Biochem Biophys Res Commun 2000 Aug 18;275(1):26-32
Phosphorylation of AZT-resistant human immunodeficiency virus type 1 reverse transcriptase
by casein kinase II in vitro: effects on inhibitor sensitivity.
Lazaro JB, Boretto J, Selmi B, Capony JP, Canard B
Department BCMP, Harvard Medical School, Boston, Massachusetts, USA.
Casein kinase II (CKII) phosphorylates wild-type (WT) recombinant reverse transcriptase (RT) mainly in the p66 subunit in vitro. Phosphorylation of T215F RT and D67N/K70R/T215F/K219Q RT (AZT-resistant RT) in vitro increases discrimination against AZTTP 2. 5- and 3.6-fold, respectively. This in vitro resistance can be reversed by treatment of phosphorylated AZT-resistant RT with phosphatase. Phosphorylation has no effect on WT RT. Terminal transferase activity of RT is selectively suppressed on phosphorylated AZT-resistant RT. Resistance to phosphonoformic acid (PFA, foscarnet) increases 3-fold upon phosphorylation of AZT-resistant RT. Although T215, the most important residue for AZT-resistance, is part of a CKII consensus target site, serines are primarily phosphorylated relative to threonines. Mutational analysis shows that phosphorylation can be reduced to 10% that of WT when amino-acid changes are introduced both in the "fingers" subdomain and motif D. These results suggest that phosphorylation of RT might be one factor involved in drug resistance in vivo. Copyright 2000 Academic Press.
PMID: 10944435, UI: 20403867
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The following is an comprehensive and excellent review of research on the Flavanoids
including Quercetin
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[12]
ASPET Journals Molecular Pharmacology Pharmacological Reviews
Journal of Pharmacology and Experimental Therapeutics Drug Metabolism and Disposition
Vol. 52, Issue 4, 673-751, December 2000
The Effects of Plant Flavonoids on Mammalian Cells:Implications for Inflammation, Heart Disease, and Cancer
Elliott Middleton, Jr., Chithan Kandaswami and Theoharis C. Theoharides1
Chebeague Island Institute of Natural Product Research, Chebeague Island, Maryland (E.M., C.K.); and Department of Pharmacology and Experimental Therapeutics, Tufts University School of Medicine, Boston, Massachusetts (T.C.T.)