RESUMO
Human immunodeficiency virus (HIV) pre-exposure prophylaxis (PrEP) strategies with proven in vivo efficacy rely on antiretroviral drugs, creating the potential for drug resistance and complicated treatment options in individuals who become infected. Moreover, on-demand products are currently missing from the PrEP development portfolio. Griffithsin (GRFT) is a non-antiretroviral HIV entry inhibitor derived from red algae with an excellent safety profile and potent activity in vitro. When combined with carrageenan (CG), GRFT has strong activity against herpes simplex virus-2 (HSV-2) and human papillomavirus (HPV) in vitro and in vivo. Here, we report that GRFT/CG in a freeze-dried fast dissolving insert (FDI) formulation for on-demand use protects rhesus macaques from a high dose vaginal SHIV SF162P3 challenge 4 h after FDI insertion. Furthermore, the GRFT/CG FDI also protects mice vaginally against HSV-2 and HPV pseudovirus. As a safe, potent, broad-spectrum, on-demand non-antiretroviral product, the GRFT/CG FDI warrants clinical development.
Assuntos
Síndrome da Imunodeficiência Adquirida/prevenção & controle , Antivirais/uso terapêutico , Carragenina/uso terapêutico , Herpes Genital/prevenção & controle , Infecções por Papillomavirus/prevenção & controle , Lectinas de Plantas/uso terapêutico , Administração Intravaginal , Animais , Antivirais/química , Carragenina/química , Modelos Animais de Doenças , Composição de Medicamentos/métodos , Avaliação Pré-Clínica de Medicamentos , Feminino , Liofilização , Herpes Genital/virologia , Herpesvirus Humano 2/patogenicidade , Humanos , Macaca mulatta , Masculino , Infecções por Papillomavirus/virologia , Lectinas de Plantas/química , Lectinas de Plantas/genética , Lectinas de Plantas/isolamento & purificação , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Profilaxia Pré-Exposição/métodos , Síndrome de Imunodeficiência Adquirida dos Símios/prevenção & controle , Vírus da Imunodeficiência Símia/patogenicidade , Nicotiana/genética , Nicotiana/metabolismo , Resultado do Tratamento , Vagina/virologiaRESUMO
Precoital, on-demand topical microbicides to reduce a woman's risk of sexually transmitted infections have been in development for nearly 3 decades, but no product has been approved due to acceptability issues and poor adherence in clinical trials. We set out to develop a self-administered vaginal fast-dissolving insert (FDI) produced by freeze-drying that would deliver safe and effective amounts of the antiviral agents griffithsin (GRFT) and carrageenan (CG) and would have properties women and their partners find acceptable. We evaluated FDI physical criteria, attributes of the gel produced upon dissolving, and GRFT stability. The lead formulation, FDI-024, was selected from 13 candidates and contains 4 mg of GRFT, 15 mg of CG, and excipients (the cryoprotectant sucrose and bulking agents dextran 40 and mannitol). The FDI exhibits good friability and hardness and is stable for at least 6 months at up to 40°C/75% relative humidity. It disintegrates in less than 60 s in a physiologically relevant volume (â¼1 mL) of simulated vaginal fluid, forming a viscous semi-solid gel with favorable mucoadhesive and spreading properties. The formulation retains the antiviral activity of GRFT and CG against HIV type 1 and human papillomavirus, respectively, in cell-based assays.
Assuntos
Antivirais/química , Antivirais/uso terapêutico , Carragenina/química , Infecções Sexualmente Transmissíveis/tratamento farmacológico , Vagina/efeitos dos fármacos , Administração Intravaginal , Excipientes/química , Feminino , Liofilização/métodos , HIV-1/efeitos dos fármacos , Humanos , Papillomaviridae/efeitos dos fármacos , SolubilidadeRESUMO
Women globally need access to multipurpose prevention technologies (MPTs) that prevent human immunodeficiency virus (HIV), sexually transmitted infections that increase HIV acquisition/transmission risk, and unintended pregnancy. Seeking an MPT with activity against HIV, herpes simplex virus-2 (HSV-2), and human papillomavirus (HPV), we developed a prototype intravaginal ring (IVR), the MZCL IVR, which released the antiviral agents MIV-150, zinc acetate, and carrageenan (MZC for short) and the contraceptive levonorgestrel (LNG). Previously, we showed that an MZC gel has potent activity against immunodeficiency viruses, HSV-2, and HPV and that the MZCL (MZC with LNG) IVR releases all four components in macaques in vivo at levels associated with efficacy. Vaginal fluid from treated macaques has in vitro activity against HIV, HSV-2, and HPV. Herein, we assessed the ability of the MZCL IVR to protect macaques against repeated co-challenge with HSV-2 and SHIV-RT (simian immunodeficiency virus [SIV] containing the reverse transcriptase gene from HIV) and prevent hormonal cycling. We evaluated in vivo drug release in co-challenged macaques by measuring drug levels in blood and vaginal fluid and residual drug levels in used IVRs. The MZCL IVR significantly prevented SHIV-RT infection, reduced HSV-2 vaginal shedding, and prevented cycling. No non-nucleoside HIV reverse transcriptase inhibitor (NNRTI)-resistant SHIV was detected in macaques that became infected after continuous exposure to MZC from the IVR. Macaques wearing the MZCL IVR also had carrageenan levels in vaginal fluid expected to protect from HPV (extrapolated from mice) and LNG levels in blood associated with contraceptive efficacy. The MZCL IVR is a promising MPT candidate that warrants further development.
Assuntos
Antivirais/administração & dosagem , Anticoncepcionais Femininos/administração & dosagem , Síndrome de Imunodeficiência Adquirida dos Símios/prevenção & controle , Cremes, Espumas e Géis Vaginais/administração & dosagem , Eliminação de Partículas Virais/efeitos dos fármacos , Alphapapillomavirus/efeitos dos fármacos , Alphapapillomavirus/fisiologia , Animais , Antivirais/farmacologia , Carragenina/administração & dosagem , Carragenina/farmacologia , Anticoncepcionais Femininos/farmacologia , Dispositivos Anticoncepcionais Femininos , Modelos Animais de Doenças , Quimioterapia Combinada/métodos , Feminino , Herpes Simples/prevenção & controle , Herpesvirus Humano 2/efeitos dos fármacos , Herpesvirus Humano 2/fisiologia , Humanos , Macaca mulatta , Ciclo Menstrual , Piridinas/administração & dosagem , Piridinas/farmacologia , Ureia/administração & dosagem , Ureia/análogos & derivados , Ureia/farmacologia , Cremes, Espumas e Géis Vaginais/farmacologia , Acetato de Zinco/administração & dosagem , Acetato de Zinco/farmacologiaRESUMO
Women need multipurpose prevention products (MPTs) that protect against sexually transmitted infections (STIs) and provide contraception. The Population Council has developed a prototype intravaginal ring (IVR) releasing the non-nucleoside reverse transcriptase inhibitor (NNRTI) MIV-150 (M), zinc acetate (ZA), carrageenan (CG) and levonorgestrel (LNG) (MZCL IVR) to protect against HIV, HSV-2, HPV and unintended pregnancy. Our objective was to evaluate the anti-SHIV-RT activity of MZCL IVR in genital mucosa. First, macaque vaginal tissues were challenged with SHIV-RT in the presence of (i) MIV-150 ± LNG or (ii) vaginal fluids (VF); available from studies completed earlier) collected at various time points post insertion of MZCL and MZC IVRs. Then, (iii) MZCL IVRs (vs. LNG IVRs) were inserted in non-Depo Provera-treated macaques for 24h and VF, genital biopsies, and blood were collected and tissues were challenged with SHIV-RT. Infection was monitored with one step SIV gag qRT-PCR or p27 ELISA. MIV-150 (LCMS/MS, RIA), LNG (RIA) and CG (ELISA) were measured in different compartments. Log-normal generalized mixed linear models were used for analysis. LNG did not affect the anti-SHIV-RT activity of MIV-150 in vitro. MIV-150 in VF from MZC/MZCL IVR-treated macaques inhibited SHIV-RT in vaginal mucosa in a dose-dependent manner (p<0.05). MIV-150 in vaginal tissue from MZCL IVR-treated animals inhibited ex vivo infection relative to baseline (96%; p<0.0001) and post LNG IVR group (90%, p<0.001). No MIV-150 dose-dependent protection was observed, likely because of high MIV-150 concentrations in all vaginal tissue samples. In cervical tissue, MIV-150 inhibited infection vs. baseline (99%; p<0.05). No cervical tissue was available for MIV-150 measurement. Exposure to LNG IVR did not change tissue infection level. These observations support further development of MZCL IVR as a multipurpose prevention technology to improve women's sexual and reproductive health.
Assuntos
Anti-Infecciosos/farmacologia , Anticoncepcionais Femininos/farmacologia , Levanogestrel/farmacologia , Piridinas/farmacologia , Vírus Reordenados/efeitos dos fármacos , Inibidores da Transcriptase Reversa/farmacologia , Ureia/análogos & derivados , Vagina/efeitos dos fármacos , Animais , Carragenina/farmacologia , Dispositivos Anticoncepcionais Femininos , Combinação de Medicamentos , Feminino , HIV/efeitos dos fármacos , HIV/genética , HIV/crescimento & desenvolvimento , Macaca mulatta , Mucosa/efeitos dos fármacos , Mucosa/virologia , Vírus Reordenados/genética , Vírus Reordenados/crescimento & desenvolvimento , Vírus da Imunodeficiência Símia/efeitos dos fármacos , Vírus da Imunodeficiência Símia/genética , Vírus da Imunodeficiência Símia/crescimento & desenvolvimento , Resultado do Tratamento , Ureia/farmacologia , Vagina/virologia , Acetato de Zinco/farmacologiaRESUMO
Nucleocapsid 7 (NCp7) inhibitors have been investigated extensively for their role in impeding the function of HIV-1 replication machinery and their ability to directly inactivate the virus. A class of NCp7 zinc finger inhibitors, S-acyl-2-mercaptobenzamide thioesters (SAMTs), was investigated for topical drug delivery. SAMTs are inherently unstable because of their hydrolytically labile thioester bond, thus requiring formulation approaches that can lend stability. We describe the delivery of N-[2-(3,4,5-trimethoxybenzoylthio)benzoyl]-ß-alaninamide (SAMT-10), as a single agent antiretroviral (ARV) therapeutic and in combination with the HIV-1 reverse-transcriptase inhibitor pyrimidinedione IQP-0528, from a hydrophobic polyether urethane (PEU) intravaginal ring (IVR) for a month. The physicochemical stability of the ARV-loaded IVRs was confirmed after 3 months at 40°C/75% relative humidity. In vitro, 25 ± 3 mg/IVR of SAMT-10 and 86 ± 13 mg/IVR of IQP-0528 were released. No degradation of the hydrolytically labile SAMT-10 was observed within the matrix. The combination of ARVs had synergistic antiviral activity when tested in in vitro cell-based assays. Toxicological evaluations performed on an organotypic EpiVaginal(™) tissue model demonstrated a lack of formulation toxicity. Overall, SAMT-10 and IQP-0528 were formulated in a stable PEU IVR for sustained release. Our findings support the need for further preclinical evaluation. © 2015 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 104:3426-3439, 2015.
Assuntos
Fármacos Anti-HIV/administração & dosagem , Fármacos Anti-HIV/uso terapêutico , Benzamidas/administração & dosagem , Benzamidas/uso terapêutico , Infecções por HIV/prevenção & controle , HIV-1/efeitos dos fármacos , Inibidores da Transcriptase Reversa/administração & dosagem , Inibidores da Transcriptase Reversa/uso terapêutico , Administração Intravaginal , Fármacos Anti-HIV/efeitos adversos , Benzamidas/efeitos adversos , Química Farmacêutica , Preparações de Ação Retardada , Análise Diferencial Térmica , Sistemas de Liberação de Medicamentos , Feminino , Infecções por HIV/transmissão , HIV-1/genética , Humanos , Técnicas de Cultura de Órgãos , Pirimidinonas/administração & dosagem , Pirimidinonas/farmacologia , SolubilidadeRESUMO
Women urgently need a self-initiated, multipurpose prevention technology (MPT) that simultaneously reduces their risk of acquiring HIV-1, HSV-2, and HPV (latter two associated with increased risk of HIV-1 acquisition) and prevents unintended pregnancy. Here, we describe a novel core-matrix intravaginal ring (IVR), the MZCL IVR, which effectively delivered the MZC combination microbicide and a contraceptive. The MZCL IVR contains four active pharmaceutical ingredients (APIs): MIV-150 (targets HIV-1), zinc acetate (ZA; targets HIV-1 and HSV-2), carrageenan (CG; targets HPV and HSV-2), and levonorgestrel (LNG; targets unintended pregnancy). The elastomeric IVR body (matrix) was produced by hot melt extrusion of the non-water swellable elastomer, ethylene vinyl acetate (EVA-28), containing the hydrophobic small molecules, MIV-150 and LNG. The solid hydrophilic core, embedded within the IVR by compression, contained the small molecule ZA and the macromolecule CG. Hydrated ZA/CG from the core was released by diffusion via a pore on the IVR while the MIV-150/LNG diffused from the matrix continuously for 94 days (d) in vitro and up to 28 d (study period) in macaques. The APIs released in vitro and in vivo were active against HIV-1ADA-M, HSV-2, and HPV16 PsV in cell-based assays. Serum LNG was at levels associated with local contraceptive effects. The results demonstrate proof-of-concept of a novel core-matrix IVR for sustained and simultaneous delivery of diverse molecules for the prevention of HIV, HSV-2 and HPV acquisition, as well as unintended pregnancy.
Assuntos
Antivirais/administração & dosagem , Dispositivos Anticoncepcionais Femininos/virologia , Sistemas de Liberação de Medicamentos/instrumentação , Infecções por HIV/prevenção & controle , Herpes Genital/prevenção & controle , Levanogestrel/administração & dosagem , Infecções por Papillomavirus/prevenção & controle , Administração Intravaginal , Animais , Antivirais/farmacocinética , Antivirais/farmacologia , Carragenina/administração & dosagem , Carragenina/farmacocinética , Carragenina/farmacologia , Linhagem Celular , Anticoncepcionais Femininos/administração & dosagem , Anticoncepcionais Femininos/farmacocinética , Anticoncepcionais Femininos/farmacologia , Desenho de Equipamento , Feminino , Transcriptase Reversa do HIV/antagonistas & inibidores , HIV-1/efeitos dos fármacos , Células HeLa , Herpesvirus Humano 2/efeitos dos fármacos , Papillomavirus Humano 16/efeitos dos fármacos , Humanos , Levanogestrel/farmacocinética , Levanogestrel/farmacologia , Macaca mulatta , Gravidez , Piridinas/administração & dosagem , Piridinas/farmacocinética , Piridinas/farmacologia , Ureia/administração & dosagem , Ureia/análogos & derivados , Ureia/farmacocinética , Ureia/farmacologia , Acetato de Zinco/administração & dosagem , Acetato de Zinco/farmacocinética , Acetato de Zinco/farmacologiaRESUMO
Tenofovir (TFV) is a nucleotide reverse transcriptase inhibitor and IQP-0528 is a non-nucleoside reverse transcriptase inhibitor that also blocks virus entry. TFV and IQP-0528 alone have shown antiviral activity as microbicide gels. Because combination therapy will likely be more potent than mono-therapy, these drugs have been chosen to make a combination microbicide gel containing 2.5% TFV/1% IQP-0528. Safety and efficacy testing was done to evaluate five prototype combination gels. The gels retained TZM-bl cell and ectocervical and colorectal tissue viability. Further, the epithelium of the ectocervical and colorectal tissue remained intact after a 24h exposure. The ED(50) calculated from the formulations for IQP-0528 was ~32nM and for TFV was ~59nM and their inhibitory activity was not affected by semen. The ED(50) of TFV in the combination gels was ~100-fold lower than when calculated for the drug substance alone reflecting the activity of the more potent IQP-0528. When ectocervical and colorectal tissue were treated with the combination gels, HIV-1 p24 release was reduced by ≥1log(10) and ≥2log(10), respectively. Immunohistochemistry for the ectocervical tissues treated with combination gels showed no HIV-1 infected cells at study end. With the increased realization of receptive anal intercourse among heterosexual couples often in conjunction with vaginal intercourse, having a safe and effective microbicide for both mucosal sites is critical. The safety and efficacy profiles of the gels were similar for ectocervical and colorectal tissues suggesting these gels have the potential for dual compartment use.
Assuntos
Adenina/análogos & derivados , Anti-Infecciosos/farmacologia , Quimioprevenção/métodos , Organofosfonatos/farmacologia , Pirimidinonas/farmacologia , Cremes, Espumas e Géis Vaginais/farmacologia , Adenina/farmacologia , Adenina/toxicidade , Administração através da Mucosa , Anti-Infecciosos/toxicidade , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Quimioterapia Combinada/métodos , Feminino , Infecções por HIV/prevenção & controle , HIV-1/efeitos dos fármacos , Humanos , Organofosfonatos/toxicidade , Pirimidinonas/toxicidade , Tenofovir , Técnicas de Cultura de Tecidos , Cremes, Espumas e Géis Vaginais/toxicidadeRESUMO
In light of the increasing worldwide AIDS pandemic, there is a continuing need to develop new prevention strategies to inhibit the transmission of HIV-1. In the absence of a successful vaccine, topical microbicides represent the best strategies to reduce the epidemic. Following the success of HIV therapeutic cocktail strategies, combinations of microbicides including nucleotide reverse transcriptase inhibitors (NtRTIs) and nonnucleoside reverse transcriptase inhibitors (NNRTIs) may offer significant protection from infection over single agents. We have developed a combination microbicide gel formulation for the delivery of IQP-0528, a novel NNRTI, and tenofovir (TFV), a NtRTI. Gel formulations were evaluated based on quantitative viscoelastic and physiochemical evaluations defined by a target product profile (TPP). For the majority of the evaluations, the gel formulations behaved similarly; all showed shear thinning behavior, were stable, nontoxic, and active against HIV-1 infection. Gel formulation F2759 displayed increased drug release of 289 ± 100 µg/(cm(2) h(1/2) ) and a tissue permeability of 60 times the half maximal effective concentration (EC(50) ) of TFV and 800 times the EC(50) of IQP-0528. In addition, F2759 showed osmolality within TPP and the highest performance in gel spreading. We have identified a gel formulation to deliver a combination microbicide of IQP-0528 and TFV that has significant potential to prevent infection of HIV-1.
Assuntos
Síndrome da Imunodeficiência Adquirida/prevenção & controle , Adenina/análogos & derivados , Fármacos Anti-HIV/administração & dosagem , Anti-Infecciosos/administração & dosagem , HIV-1/efeitos dos fármacos , Organofosfonatos/administração & dosagem , Pirimidinonas/administração & dosagem , Inibidores da Transcriptase Reversa/administração & dosagem , Adenina/administração & dosagem , Adenina/química , Anti-Infecciosos/química , Linhagem Celular , Química Farmacêutica , Excipientes , Géis , Humanos , Organofosfonatos/química , Solubilidade , Tenofovir , ViscosidadeRESUMO
Pyrimidinediones, a novel class of compounds, have previously been shown to possess antiviral activity at nanomolar concentrations. One member of this class of compounds, IQP-0528, was selected as the lead molecule for formulation development owing to its stability at physiologically relevant conditions, wide therapeutic window, and antiviral activity in the nanomolar range. Here, we report the development of two vaginal gels--3.0% hydroxyethyl cellulose (HEC) formulation and a 0.65% Carbopol formulation--for the sustained delivery of IQP-0528. Stability studies under accelerated conditions confirmed the chemical stability of IQP-0528 and mechanical stability of the gel formulation for 3 months. In vitro release studies revealed that diffusion-controlled release of IQP-0528 occurred over 6 h, with an initial lag time of approximately 1 h. Based on the drug release profile, the 3.0% HEC gel was selected as the lead formulation for safety and activity evaluations. The in vitro and ex vivo safety evaluations showed no significant loss in cell viability or significant inflammatory response after treatment with a 3.0% HEC gel containing 0.25% IQP-0528. In an in vitro HIV-1 entry inhibition assay, the lead formulation showed an 50% effective concentration of 0.14 µg/ml for gel in culture media, which corresponds to â¼0.001 µM IQP-0528. The antiviral activity was further confirmed by using polarized cervical explants, in which the formulation showed complete protection against HIV infection. In summary, these results are encouraging and warrant further evaluation of IQP-0528 gel formulations in in vivo models, as well as the development of alternative formulations for the delivery of IQP-0528 as a microbicide.
Assuntos
Fármacos Anti-HIV/administração & dosagem , Fármacos Anti-HIV/farmacologia , Pirimidinonas/administração & dosagem , Pirimidinonas/farmacologia , Cremes, Espumas e Géis Vaginais/administração & dosagem , Animais , Fármacos Anti-HIV/química , Feminino , HIV-1/efeitos dos fármacos , Humanos , Técnicas In Vitro , Pirimidinonas/química , Suínos , Vagina/metabolismo , Cremes, Espumas e Géis Vaginais/químicaRESUMO
Supercritical carbon dioxide (scCO(2)) processing of drug/polymer mixtures is an environmentally friendly means of creating an impregnated polymeric carrier to enhance the aqueous dissolution rate of drugs that exhibit poor water solubility or are thermally labile. However, the role of drug solubilization and its interaction with the polymer during scCO(2) processing on the extent and rate of dissolution has been ambiguous. In this study, we examine the rate of dissolution of carbamazepine (CBZ), a hydrophobic drug for treating epilepsy, in scCO(2) (90-200bar, 35°C and 45°C) and its partitioning into polyvinylpyrrolidone (PVP, 10 and 29K MW) using in situ UV-vis spectroscopy. Our results show that partitioning occurs by surface adsorption and impregnation within the polymer matrix. These processes are linked to plasticization, which is dependent on PVP molecular weight, and temperature and pressure during treatment. The rate and extent of CBZ solubility is also controlled by treatment condition. The ability to tune polymer and drug simultaneously can be used to control the nature and extent of drug loading.
Assuntos
Anticonvulsivantes/química , Carbamazepina/química , Dióxido de Carbono/química , Portadores de Fármacos/química , Povidona/química , Química Farmacêutica , Composição de Medicamentos , Interações Hidrofóbicas e Hidrofílicas , Microscopia Eletrônica de Varredura , Estrutura Molecular , Solubilidade , Soluções , Espectrofotometria Ultravioleta , Propriedades de SuperfícieRESUMO
A number of studies have been carried out to investigate the crystalline to amorphous conversion of carbamazepine (CBZ) using solid dispersion techniques. In this study we have tried to achieve conversion using a novel technique combining near-supercritical carbon dioxide (n-scCO(2)) and pharmaceutically acceptable polymers (Na CMC and PVP) of varying molecular weights. Physical mixtures were prepared in two identical sets, one exposed to n-scCO(2) treatment and other was untreated. The treated physical mixtures were compared to untreated using PXRD, DSC and USP in vitro dissolution techniques. Routinely used PXRD analysis involves qualitative estimation of the amorphous conversion of a drug. In this work a previously developed mathematical parameter, ratio of ratios (ROR), was utilized to better quantify the crystalline to amorphous conversion of CBZ. The findings from the three methods indicated that only the lowest molecular weight PVP, PVP10k, facilitated significant crystalline to amorphous conversion of CBZ. In vitro dissolution, which is considered as an estimate of bioavailability demonstrated an initial dissolution of CBZ significantly greater in the treated physical mixtures of PVP10k:CBZ than the initial dissolution of the corresponding untreated physical mixtures and pure untreated CBZ.