betadex and HIV-Infections

This randomized, open-label, comparative study assessed the pharmacokinetics and safety of intravenous and oral hydroxypropyl-beta-cyclodextrin (HP-beta-CD) solutions of itraconazole in patients with advanced human immunodeficiency virus (HIV) infection. All patients received 1-hour intravenous infusions of itraconazole 200 mg twice dailyfor 2 days, then once dailyfor 5 days. Patients were then randomized to receive itraconazole oral solution, 200 mg twice daily or 200 mg once daily, for a further 28 days. Itraconazole was solubilized by HP-beta-CD in both intravenous and oral solutions, so HP-beta-CD concentration in plasma was measured. Thirty-two patients were enrolled and analyzed (n = 32 for intravenous treatment, 32 completed; n = 16 for oral once daily, 15 completed; n = 16 for oral twice daily, 12 completed). Steady-state plasma concentrations of itraconazole and hydroxyitraconazole were reached by days 3 and 6, respectively. After intravenous dosing, mean trough plasma concentrations of itraconazole and hydroxyitraconazole were 906 ng/ml and 1,690 ng/ml, respectively. During oral dosing, mean trough plasma concentrations of itraconazole and hydroxyitraconazole were maintained or increased in the 200 mg twice-dailygroup but fell with the 200 mg once-daily oral dose. Itraconazole was generally well tolerated and had a favorable safetyprofile; minor changes in hematology variables were noted during the intravenous phase, and HP-beta-CD was cleared rapidly, mostly in urine. Twenty-eight patients (88%) experienced at least one adverse event; no adverse event was severe, and only seven were definitely related to itraconazole. In conclusion, itraconazole 200 mg given intravenously twice daily for 2 days, then once daily for 5 days, rapidly achieves amean steady-state trough concentration of itraconazole of over 250 ng/ml, which is associated with clinic outcome and is effectively maintained with itraconazole oral solution 200 mg twice daily in patients with advanced HIV infection.

Topics: 2-Hydroxypropyl-beta-cyclodextrin; Administration, Oral; Adult; Antifungal Agents; Area Under Curve; beta-Cyclodextrins; Biotransformation; Cyclodextrins; Female; HIV Infections; Humans; Injections, Intravenous; Itraconazole; Male

Nowadays, acquired immunodeficiency syndrome has become a formidable danger to human health, and its early diagnosis is urgent need with the increasing quantity of patients around the world. Herein, we first synthesized beta-cyclodextrin-functionalized CdS nanorods (β-CD@CdS NRs) with high stability and desirable photo-electricity activity, and served as easy-to-assemble building modules to design a novel photoelectrochemical biosensor for human immune deficiency virus (HIV) DNA detection by coupling with catalytic hairpin assembly (CHA)-mediated biocatalytic precipitation and the host-guest interaction between adamantine (ADA) and β-CD. In the presence of HIV DNA, CHA process was triggered with the aid of hairpin DNA1 and ADA-labelled hairpin DNA2, and then generated large amounts of G-quadruplex, which could be formed hemin/G-quadruplex DNAzyme to catalyze 4-chloro-1-naphthol to generate insoluble precipitation on photoelectrode surface, followed by the decreased photocurrent response due to the corresponding stereo-hindrance effect. Under optimized conditions, this biosensor exhibited wide linear dynamic range (10 fM - 1 nM) and low detection limit of 1.16 fM, as well as high sensitivity, excellent stability, and satisfactory feasibility in human-serum samples. Moreover, the prepared β-CD@CdS NRs could be applied to the construction of other advanced sensing platform, showing great prospect in clinical diagnostics.

Topics: beta-Cyclodextrins; Biosensing Techniques; Cadmium Compounds; DNA, Catalytic; DNA, Viral; Electrochemical Techniques; G-Quadruplexes; HIV; HIV Infections; Humans; Limit of Detection; Nanotubes; Sulfides

The purpose of this work was to design and evaluate a vaginal film delivery system for UAMC01398, a novel non-nucleoside reverse transcriptase inhibitor currently under investigation for use as an anti-HIV microbicide. UAMC01398 (1mg) films consisting of hydroxypropylmethylcellulose (HPMC) and polyethylene glycol 400 (PEG400) in different ratios were prepared by solvent evaporation. Based on its flexibility, softness and translucent appearance, the 30% PEG400 and 70% HPMC containing film was selected for further assessment. The vaginal film formulation was fast-dissolving (<10 min in 1 mL of vaginal fluid simulant), stable up to at least one month and safe toward epithelial cells and lactobacilli. Furthermore, formulating UAMC01398 into the film dosage form did not influence its antiviral activity. Powder X-ray diffraction revealed the amorphous nature of the UAMC01398 film, resulting in enhanced compound permeation across the epithelial HEC-1A cell layer, presumably owing to the induction of supersaturation. The in vivo vaginal tissue uptake of UAMC01398 in rabbits, as measured by systemic concentrations, was increased compared to the previously established non-solubilizing gel (significant difference) and sulfobutyl ether-β-cyclodextrin (5%) containing gel. To conclude, we identified a film formulation suitable for the vaginal delivery of UAMC01398.

Topics: Administration, Intravaginal; Animals; Anti-HIV Agents; beta-Cyclodextrins; Chemistry, Pharmaceutical; Drug Delivery Systems; Epithelial Cells; Excipients; Female; HIV Infections; Humans; Hypromellose Derivatives; Lactobacillus; Polyethylene Glycols; Rabbits; Reverse Transcriptase Inhibitors; Solvents; Vagina; Vaginal Creams, Foams, and Jellies

In human trophoblastic cells, a correlation between early endosomal trafficking of HIV-1 and virus infection was previously documented. However, if HIV-1 is massively internalized in these cells, the endocytic pathway(s) responsible for viral uptake is still undefined. Here we address this vital question. Amongst all the putative endocytic pathways present in polarized trophoblastic cells, we demonstrate that HIV-1 infection of these cells is independent of clathrin-mediated endocytosis and macropinocytosis. Importantly, treatment with the cholesterol-sequestering drug filipin severely impairs virus internalization, whereas the cholesterol-depleting compound methyl-beta-cyclodextrin has no impact on this pathway. Moreover, viral internalization is unaffected by overexpression of a mutant dynamin 2 or treatment with a kinase or tyrosine phosphatase inhibitor. Thus, HIV-1 infection in polarized trophoblastic cells occurs primarily via a clathrin, caveolae, and dynamin-independent pathway requiring free cholesterol. Notably, even though HIV-1 did not initially co-localize with transferrin, some virions migrate at later time points to transferrin-enriched endosomes, suggesting an unusual transit from the non-classical pathway to early endosomes. Finally, virus internalization in these cells does not involve the participation of microtubules but relies partly on actin filaments. Collectively these findings provide unprecedented information on the route of HIV-1 internalization in polarized human trophoblasts.

Topics: Actins; Adult; beta-Cyclodextrins; Caveolae; Cell Line; Cell Membrane; Cell Polarity; Child; Cholesterol; Clathrin; Cytoskeleton; Dynamins; Endocytosis; Female; Filipin; HIV Infections; HIV-1; Humans; Membrane Microdomains; Microtubules; Transferrin; Trophoblasts; Tumor Necrosis Factor-alpha; Virus Internalization

Cervical tissue-based organ culture system has been used to test the cytotoxicity and antiviral activity of microbicides. One of the problems of using current organ culture methods for routine microbicide testing is the need to continually obtain fresh tissue, which can be limited in access and supply. Use of frozen tissue, stored when available and thawed when needed, would alleviate the need for constant access to new tissue. This study was designed to explore the possibility of using frozen-thawed cervical tissue to test microbicides for their anti-HIV activity. We provided biochemical, histological, and quantitative immunohistochemical data to demonstrate the integrity of the frozen-thawed organ culture system. Significant levels of HIV-1 mucosal transmission were noted with both fresh and frozen-thawed tissue, regardless of the coreceptor usage of the virus isolate. Furthermore, candidate microbicides UC781, beta-cyclodextrin, and octylglycerol inhibited HIV-1 transmission across the mucosa of frozen-thawed tissues with a level of efficiency similar to that of fresh tissues. Therefore, frozen-thawed cervical tissue in the organ culture system provides a practical and convenient model to screen topical microbicides for their ability to block sexual transmission of HIV-1, and reduces the problems associated with procurement of the numerous tissues required for evaluation and comparison of microbicide candidates among different laboratories.

Topics: Anilides; Anti-Infective Agents; Anti-Retroviral Agents; beta-Cyclodextrins; Cervix Uteri; Female; Freezing; Furans; Glycerol; HIV Infections; HIV-1; Humans; Middle Aged; Organ Culture Techniques; Temperature; Thioamides

The intracellular activity of certain antiviral agents, including antisense oligonucleotides, acyclic nucleoside phosphonates, and protease inhibitors, is enhanced when they are delivered in liposome-encapsulated form. In this chapter we describe the preparation of pH-sensitive liposomes encapsulating antisense oligonucleotides, ribozymes, and acyclic nucleoside phosphonate analogues and their effects on HIV replication in macrophages. We outline the use of liposomal HIV protease inhibitors in infected macrophages. We present two methods for the covalent coupling of soluble CD4 to liposomes and show the association of these liposomes with HIV-infected cells. We also describe the synthesis of a novel antiviral agent based on cyclodextrin and its incorporation into liposomes.

Topics: Antiviral Agents; beta-Cyclodextrins; CD4 Antigens; Cross-Linking Reagents; Drug Delivery Systems; HIV Envelope Protein gp120; HIV Infections; HIV-1; Humans; Hydrogen-Ion Concentration; Liposomes; Macrophages; Microbial Sensitivity Tests; Oligonucleotides, Antisense; Organophosphonates; Protease Inhibitors; Purines; Pyrimidines; RNA, Catalytic; Succinimides; Sulfides

We have shown that HIV budding occurs at cholesterol-rich membrane microdomains called lipid rafts (Nguyen and Hildreth, J Virol 2000;74:3264-3272). This observation prompted us to examine the role in HIV entry of cholesterol in the membrane of cells. We recently reported that host cell cholesterol is required for HIV infection (Liao et al., AIDS Res Hum Retroviruses 2001;17:1009-1019). In the present study we examined the role of virion-associated cholesterol in HIV infection by modulating the cholesterol content of virions and infected cells with 2-hydoxypropyl-beta-cyclodextrin (beta-cyclodextrin). Our results show that removal of cholesterol from the membrane of HIV-infected cells dramatically lowered virus release and that virions released from cholesterol-depleted cells are minimally infectious. Exposure of infectious HIV particles to beta-cyclodextrin resulted in a dose-dependent inactivation of the virus. In both cases, the effect was attributable to loss of cholesterol and could be reversed by replenishing cholesterol. beta-Cyclodextrin-treated, noninfectious HIV retained its ability to bind cells. Western blot, p24 core ELISA, and reverse transcription assays indicated that virions remained intact after treatment with beta-cyclodextrin at concentrations that abolished infectivity. Electron microscopy revealed that beta-cyclodextrin-treated HIV had a morphology very similar to that of untreated virus. R18 fluorescence dequenching studies showed that beta-cyclodextrin-treated HIV did not fuse to the membrane of susceptible cells. Dequenching was restored by replenishing virion-associated cholesterol. The results indicate that cholesterol in HIV particles is strictly required for fusion and infectivity. These observations in combination with those of past studies indicate beta-cyclodextrin to be an excellent candidate for use as a chemical barrier for AIDS prophylaxis.

Topics: 2-Hydroxypropyl-beta-cyclodextrin; beta-Cyclodextrins; Blotting, Western; Cell Line; Cholesterol; Cyclodextrins; HIV Infections; HIV-1; Humans; Virion

Because both HIV-1 virions and HIV-infected cells are present in the semen and cervical mucus of infected individuals, HIV-1 prevention strategies must consider both cell-free and cell-associated virus. Antibodies that target HIV-1 virions have been shown to prevent vaginal transmission of cell-free virus in macaques, but since cell-associated transmission has not been reliably demonstrated in this model system, no strategies to prevent such transmission have been tested. We have employed a mouse model in which SCID mice carry human peripheral blood leukocytes (HuPBLs). In these mice, vaginal transmission of cell-associated, but not cell-free, HIV-1 transmission occurs, mediated by transepithelial migration of HIV-infected cells. Topical application of beta-cyclodextrin (beta-CD), a cholesterol-sequestering agent that interferes with cell migration and budding of virus from lipid rafts, blocks transmission of cell-associated HIV-1. The HuPBL-SCID model of vaginal HIV-1 transmission should prove useful for investigating cell-associated HIV-1 transmucosal HIV-1 transmission, as well as for screening reagents for their potential efficacy in preventing sexual HIV-1 transmission.

Topics: 2-Hydroxypropyl-beta-cyclodextrin; Administration, Topical; Animals; beta-Cyclodextrins; Cell Movement; Cyclodextrins; Disease Models, Animal; Epithelium; Excipients; Female; HeLa Cells; HIV Infections; HIV-1; Humans; Leukocytes, Mononuclear; Lymph Nodes; Mice; Mice, SCID; Peritoneal Cavity; Progesterone; Vagina

HIV requires cholesterol and lipid rafts on target cell membranes for infection. To elucidate a possible mechanism, we determined that cholesterol extraction by hydroxypropyl-beta-cyclodextrin (BCD) inhibits stromal cell-derived factor 1alpha (SDF-1alpha) binding to CXCR4 on T cell lines and PBMCs. Intracellular calcium responses to SDF-1alpha, as well as receptor internalization, were impaired in treated T cells. Loss in ligand binding is likely due to conformational changes in CXCR4 and not increased sensitivity to internalization. SDF-1alpha binding and calcium responses were effectively restored by reloading cholesterol. Immunofluorescence microscopy revealed that SDF-1alpha binding occurred in lipid raft microdomains that contained GM1. CXCR4 surface expression, on the other hand, only partially colocalized with GM1. HIV-1(IIIB) infection assays confirmed the functional loss of CXCR4 in the cell lines tested, Sup-T1 and CEM-NKR-CCR5. These data suggest that cholesterol is essential for CXCR4 conformation and function and that lipid rafts may play a regulatory role in SDF-1alpha signaling.

Topics: 2-Hydroxypropyl-beta-cyclodextrin; Anti-HIV Agents; Antibodies, Monoclonal; beta-Cyclodextrins; Binding Sites, Antibody; Calcium; Cell Membrane; Chemokine CXCL12; Chemokines, CXC; Cholesterol; Clone Cells; Cyclodextrins; Excipients; HIV Infections; HIV-1; Humans; Intracellular Fluid; Jurkat Cells; Membrane Microdomains; Protein Binding; Receptors, CXCR4; T-Lymphocytes

In a previous study we showed that budding of HIV-1 particles occurs at highly specialized membrane microdomains known as lipid rafts. These microdomains are characterized by a distinct lipid composition that includes high concentrations of cholesterol, sphingolipids, and glycolipids. Since cholesterol is known to play a key role in the entry of some other viruses, our observation of HIV budding from lipid rafts led us to investigate the role in HIV-1 entry of cholesterol and lipid rafts in the plasma membrane of susceptible cells. We have used 2-OH-propyl-beta-cyclodextrin (beta-cyclodextrin) to deplete cellular cholesterol and disperse lipid rafts. Our results show that removal of cellular cholesterol rendered primary cells and cell lines highly resistant to HIV-1-mediated syncytium formation and to infection by both CXCR4- and CCR5-specific viruses. beta-Cyclodextrin treatment of cells partially reduced HIV-1 binding, while rendering chemokine receptors highly sensitive to antibody-mediated internalization. There was no effect on CD4 expression. All of the above-described effects were readily reversed by incubating cholesterol-depleted cells with low concentrations of cholesterol-loaded beta-cyclodextrin to restore cholesterol levels. Cholesterol depletion made cells resistant to SDF-1-induced binding to ICAM-1 through LFA-1. Since LFA-1 contributes significantly to cell binding by HIV-1, this latter effect may have contributed to the observed reduction in HIV-1 binding to cells after treatment with beta-cyclodextrin. Our results indicate that cholesterol may be critical to the HIV-1 coreceptor function of chemokine receptors and is required for infection of cells by HIV-1.

Topics: beta-Cyclodextrins; Cell Line; Cholesterol; Cyclodextrins; HIV Infections; HIV-1; Humans; Jurkat Cells; Membrane Microdomains; Protein Binding; Receptors, CCR5; Receptors, CXCR4; Tumor Cells, Cultured

Topics: Acetates; Aged; AIDS Vaccines; American Medical Association; Amines; Animals; Anticonvulsants; beta-Cyclodextrins; Cells, Cultured; Clinical Trials as Topic; Cochlear Implants; Complementary Therapies; Constriction, Pathologic; Coronary Artery Bypass; Cyclodextrins; Cyclohexanecarboxylic Acids; Depressive Disorder; Drug Therapy, Combination; Epilepsy; Extracorporeal Membrane Oxygenation; Felbamate; Fetus; Gabapentin; gamma-Aminobutyric Acid; Genetic Therapy; Health Care Costs; HIV Infections; Humans; Hyperlipoproteinemia Type II; Lamotrigine; Muscle, Smooth; National Institutes of Health (U.S.); Oxygen; Phenylcarbamates; Propylene Glycols; Respiration, Artificial; Respiratory Distress Syndrome; Triazines; United States; Vascular Surgical Procedures