Cholesterol-conjugated poly(D,L-lactide)-based micelles as a nanocarrier system for effective delivery of curcumin in cancer therapy

Polymeric micelles have been widely explored preclinically as suitable delivery systems for poorly soluble chemotherapeutic drugs in cancer therapy. The present study reported the development of cholesterol (Ch)-conjugated poly(D,L-Lactide) (PLA)-based polymeric micelles (mPEG–PLA-Ch) for effective encapsulation and delivery of curcumin (CUR) at the tumor site. Cholesterol conjugation dramatically affected the particle size and improved drug loading (DL) and encapsulation efficiency (EE). mPEG–PLA-Ch-CUR showed bigger hydrodynamic diameter (104.6?±?2.1?nm, and 169.3?±?1.52?nm for mPEG–PLA and mPEG–PLA-Ch, respectively) due to increased size of the hydrophobic core. The newly developed polymer exhibited low critical micelles concentration (CMC) (25?μg/mL) which is close to lipid-based polymer, PEG-phosphatidyl ethanolamine (12.5?μg/mL) compared to mPEG–PLA (50?μg/mL). mPEG–PLA-Ch micelles exhibited relatively higher EE (93.74?±?1.6%) and DL (11.86?±?0.8%) compared to mPEG–PLA micelles (EE 91.89?±?1.2% and DL 11.06?±?0.8%). mPEG–PLA-Ch micelles were internalized by the cancer cells effectively and exhibited higher cytotoxicity compared to free CUR in both, murine melanoma (B16F10) and human breast cancer (MDA-MB-231) cells. mPEG–PLA-Ch exhibited satisfactory hemocompatibility indicating their potential for systemic application. Further, mPEG–PLA-Ch-CUR demonstrated higher rate of reduction of tumor volume in B16F10-xenografted tumor-bearing mice compared to free CUR. At the end of 22 days, the tumor reduced to 1.87-fold (627.72?±?0.9?mm³ versus 1174.68?±?1.64?mm³) compared to the treatment with free CUR. In conclusion, the experimental data in vitro and in vivo indicated that the newly developed CUR-mPEG–PLA-Ch micelles may have promising applications in solid tumors.     

Impact of Method of Preparation of Amorphous Solid Dispersions on Mechanical Properties: Comparison of Coprecipitation and Spray Drying

Usage of the amorphous phase of compounds has become the method of choice to overcome oral bioavailability problems related to poor solubility. Due to the unstable nature of glasses, it is clear that the method of preparation of the amorphous glass will have an impact on physical/chemical stability and in turn in vivo performance. The method of preparation can also have a profound impact on the mechanical properties of the amorphous phase. We have explored the impact of preparation method on the mechanical properties of an amorphous solid dispersion using a development compound, GDC-0810. Three methods were used to generate amorphous solid dispersions (ASDs) of 50% GDC-0810 with hydroxypropyl methylcellulose acetate succinate: (1) spray drying, (2) coprecipitation using overhead mixing, and (3) coprecipitation using resonant acoustic mixing. All 3 methods were found to generate ASDs with good phase mixing and similar glass transition temperatures. Coprecipitated ASD powders (overhead mixing and resonant acoustic mixing) demonstrated superior tabletability and flow properties when compared to the spray drying powder. Careful choice of manufacturing process can be used to tune material properties of ASDs to make them more amenable for downstream operations like tableting. Acoustic mixing has been demonstrated as a scalable new method to make ASDs through coprecipitation.     

Effect of pH and Excipients on Structure, Dynamics, and Long-Term Stability of a Model IgG1 Monoclonal Antibody upon Freeze-Drying

Purpose

To investigate the mechanism of IgG1 mAb stabilization after freeze-drying and the interdependence of protein structural preservation in the solid state, glassy state dynamics and long-term storage stability under different formulation conditions.

Methods

IgG1 mAb was formulated with mannitol at pH 3.0, 5.0, and 7.0 in the presence and absence of sucrose and stability was monitored over 1 year at different temperatures. Physical and covalent degradation of lyophilized formulation was monitored using SEC, CEX, and light obscuration technique. Secondary and tertiary structure of the protein in the solid state was characterized using FTIR and fluorescence spectroscopy respectively. Raman spectroscopy was also used to monitor changes in secondary and tertiary structure, while SS-NMR ¹H relaxation was used to monitor glassy state dynamics.

Results

IgG1 mAb underwent significant secondary structural perturbations at pH 3.0 and conditions without sucrose, while pH 5.0 condition with sucrose showed the least structural change over time. The structural changes correlated with long-term stability with respect to protein aggregate formation and SbVP counts. SS-NMR data showed reduced relaxation time at conditions that were more stable.

Conclusions

Native state protein structural preservation and optimal solid-state dynamics correlate with improved long-term stability of the mAb in the different lyophilized formulations.     

Oral Disintegration Tablets of Stavudine for HIV Management: A New Technological Approach

Stavudine oral disintegration tablets were formulated to minimize the bitter taste and to reduce the first-pass hepatic metabolism. The various precompression parameters like the angle of repose, bulk density, compressibility index and Hausner''s ratio were determined for the powder blend. In this study, 14 formulations of stavudine oral disintegration tablet were prepared by direct compression method. The tablets were evaluated for weight variation, percentage friability, disintegration time, hardness, wetting time and water absorption ratio. The in vitro dissolution study results of the batch S1 (stavudine+crospovidone+sodium starch glycollate) are encouraging as highest dissolution rate (99.2% in 100 min) and lowest time of disintegration (56 s) was achieved. The in vivo drug release studies were carried out in rabbits and the relative bioavailability of formulation S1 was found to be 2.83 times greater than that of conventional tablets.     

Changes in Tdap and MCV4 Vaccine Coverage Following Enactment of a Statewide Requirement of Tdap Vaccination for Entry Into Sixth Grade

Objectives. We evaluated changes in tetanus toxoid, reduced diptheria toxoid, acellular pertussis (Tdap), and tetravalen meningococcal (MCV4) vaccine coverage following enactment of a New York State mandate requiring Tdap before entering sixth grade.Methods. Using data from a hospital-based immunization registry, we measured Tdap and MCV4 coverage among youths aged 11 to 14 years in New York City at 3 time points: premandate, mandate year 1, and mandate year 2.Results. Among overlapping cohorts of 4316 (premandate), 4131 (mandate year 1), and 3639 (mandate year 2) youths, Tdap coverage increased steadily over time (29%, 58%, and 83%, respectively). Increases were observed among all ages. Across the same time points, MCV4 coverage also increased (10%, 30%, and 60%, respectively). Most adolescents did not receive MCV4 during the same visit they received Tdap.Conclusions. A Tdap school-entry mandate was associated with substantial increases in immunization coverage, even in age groups not directly affected by the mandate. At the postmandate time points, MCV4 coverage remained lower than Tdap coverage. Provider education should emphasize the importance of reviewing vaccine records and administering all recommended vaccines at every clinical encounter.In recent years, new vaccines against pertussis¹ and meningitis² have been introduced to the routine immunization schedule for children and adolescents. Both are recommended for youths aged 11 to 12 years, with catch-up vaccination for older adolescents. Pertussis is a highly contagious infection, often causing school or community outbreaks. Among healthy adolescents, pertussis is usually a self-limited illness characterized by a prolonged cough. However, secondary complications can occur, and adolescents serve as an important reservoir for transmission to infants, for whom infection can lead to pneumonia, respiratory failure, apnea, and even death.³ The tetanus toxoid, reduced diphtheria toxoid, and acellular pertussis (Tdap) vaccine has been shown to be 92% effective in preventing culture-confirmed pertussis.⁴Adolescents, and specifically those in crowded living conditions, have been shown to be at increased risk for N. meningitides infection.⁵ N. meningitides is highly contagious and can cause meningitis, septicemia, and death. The tetravalent meningococcal polysaccharide-protein conjugate (MCV4) vaccine has been shown to be safe and highly immunogenic in protecting against N. meningitides infection.⁶Although the Tdap and MCV4 vaccines hold great promise, achieving high immunization coverage among adolescents remains a challenge. Barriers to adolescent immunization include failure to present for medical services, missed immunization opportunities, and scattered immunization records.⁷ Few adolescents report the receipt of annual preventive health visits,⁸ so reviewing immunization status and immunizing at every clinical encounter is key to increasing vaccine coverage in this population. Results of the 2008 National Immunization Survey–Teen indicate that among adolescents aged 13 to 17 years, nationwide coverage for Tdap and MCV4 remains low (41% and 42%, respectively).⁹Mandates requiring immunization prior to school entry have been highly effective in increasing immunization coverage.^10–14 In 1 study, hepatitis B immunization rates increased from 13% to 71% following implementation of a middle school mandate.¹⁵ Mandates were initially used to promote the uptake of vaccines for highly contagious infectious diseases and thus to prevent school-based outbreaks, but today many states mandate vaccines for diseases that are not communicable (tetanus) or that are communicable primarily through sexual or blood exposures (hepatitis B). Recent controversy regarding mandates for the human papillomavirus vaccine has resulted in significant backlash against mandates,^16–18 highlighting the need for states to be judicious in their decisions to implement new mandates.Although mandates are known to rapidly increase vaccine coverage in their target population, there is little evidence of any carryover benefits. No studies to date have evaluated whether a new mandate will result in improved vaccine coverage for nonmandated age groups or for other nonmandated, age-appropriate vaccines. In the fall of 2007, New York State became one of the first states to require Tdap prior to entering sixth grade. This situation provided a unique opportunity to observe postmandate changes in coverage for Tdap (the mandated vaccine) and MCV4 (a nonmandated, recommended vaccine) across multiple age groups.     

Effect of Microenvironmental pH Modulation on the Dissolution Rate and Oral Absorption of the Salt of a Weak Acid – Case Study of GDC-0810

Purpose

The purpose of this work is to investigate the effect of microenvironmental pH modulation on the in vitro dissolution rate and oral absorption of GDC-0810, an oral anti-cancer drug, in human.

Methods

The pH-solubility profile of GDC-0810 free acid and pH_max of its N-Methyl-D-glucamine (NMG) salt were determined. Precipitation studies were conducted for GDC-0810 NMG salt at different pH values. GDC-0810 200-mg dose NMG salt tablet formulations containing different levels of sodium bicarbonate as the pH modifier were tested for dissolution under the dual pH-dilution scheme. Three tablet formulations were evaluated in human as a part of a relative bioavailability study. A 200-mg dose of GDC-0810 was administered QD with low fat food.

Results

Intrinsic solubility of GDC-0810 free acid was found to be extremely low. The pH_max of the NMG salt suggested a strong tendency for form conversion to the free acid under GI conditions. In vitro dissolution profiles showed that the dissolution rate and extent of GDC-0810 increased with increasing the level of sodium bicarbonate in the formulation. The human PK data showed a similar trend for the geometric mean of C_max and AUC_0-t for formulations containing 5%, 10%, and 15% sodium bicarbonate, but the difference is not statistically significant.

Conclusion

Incorporation of a basic pH modifier, sodium bicarbonate, in GDC-0810 NMG salt tablet formulations enhanced in vitro dissolution rate of GDC-0810 via microenvironmental pH modulation. The human PK data showed no statistically significant difference in drug exposure from tablets containing 5%, 10%, and 15% sodium bicarbonate.

     

Population pharmacokinetics of lamotrigine in Indian epileptic patients

Purpose

The aim of this analysis was to describe the pharmacokinetics of oral lamotrigine (LTG) in Indian epileptic patients using a population pharmacokinetic (PPK) modeling approach to confirm that the PK is similar to that of the Caucasian population, and to evaluate and confirm the impact of covariates predictive of inter-individual variability using a simulation platform.

Methods

Blood samples were obtained from 95 patients, and LTG plasma concentrations were determined. Population PK modeling was performed using NONMEM. A one-compartment PK model with first-order absorption and elimination was used to describe the LTG PK. Log-likelihood profiling and normalized prediction distribution errors (NPDE) were used for model evaluation. A simulation study was performed to investigate dose regimens.

Results

Clearance (CL) was estimated to be 2.27 L/h with inter-individual variability (IIV) of 29 CV%. Volume of distribution (V) was estimated to be 53.6 L (31 CV% IIV). Body weight and concurrent use of carbamazepine and valproate were identified as significant covariates on clearance. Log-likelihood profiling indicated that parameters could be estimated with adequate precision, and NPDE indicated that the model adequately described the data observed. The simulation study illustrated the impact of carbamazepine and valproate on LTG PK, and negligible differences in PK between Indian and Caucasian patients.

Conclusions

This is the first PK analysis of LTG in Indian patients. The population PK model developed adequately described the data observed. Comparison of identified PK parameters with previous PK analyses in Caucasian patients indicates that CL of LTG is similar, while V is somewhat lower compared with Caucasian patients, but this is not expected to lead to relevant differences in PK profiles during steady state.     

Phosphatidylserine containing liposomes reduce immunogenicity of recombinant human factor VIII (rFVIII) in a murine model of hemophilia A

Factor VIII (FVIII) is a multidomain protein that is deficient in hemophilia A, a clinically important bleeding disorder. Replacement therapy using recombinant human FVIII (rFVIII) is the main therapy. However, approximately 15-30% of patients develop inhibitory antibodies that neutralize rFVIII activity. Antibodies to epitopes in C2 domain, which is involved in FVIII binding to phospholipids, are highly prevalent. Here, we investigated the effect of phosphatidylserine (PS)-containing liposomes, which bind to C2 domain with high affinity and specificity, upon the immunogenicity of rFVIII. Circular dichroism studies showed that PS-containing liposomes interfered with aggregation of rFVIII. Immunogenicity of free- versus liposomal-rFVIII was evaluated in a murine model of hemophilia A. Animals treated with s.c. injections of liposomal-rFVIII had lower total- and inhibitory titers, compared to animals treated with rFVIII alone. Antigen processing by proteolytic enzymes was reduced in the presence of liposomes. Animals treated with s.c. injections of liposomal-rFVIII showed a significant increase in rFVIII plasma concentration compared to animals that received rFVIII alone. Based on these studies, we hypothesize that specific molecular interactions between PS-containing bilayers and rFVIII may provide a basis for designing lipidic complexes that improve the stability, reduce the immunogenicity of rFVIII formulations, and permit administration by s.c. route.     

Communication Competence,Social Support,and Depression Among College Students: A Model of Facebook and Face-to-Face Support Network Influence

Given the extensive use of the Internet for health information, Web-based health promotion interventions are widely perceived as an effective communication channel. The authors conducted this study to determine use of a Web-based intervention intended to improve colorectal cancer screening in a population of women who are at average risk and noncompliant to current screening recommendations. The study was a randomized controlled trial designed to compare the effectiveness of colorectal cancer screening educational materials delivered using the Internet versus a printed format. In 3 years, 391 women seen for routine obstetrics/gynecology follow-up at 2 academic centers provided relevant survey information. Of these, 130 were randomized to the Web intervention. Participants received voluntary access to a password-protected, study-specific Web site that provided information about colorectal cancer and colorectal cancer screening options. The main outcome measures were self-reported and actual Web site use. Only 24.6% of women logged onto the Web site. Age was the only variable that differentiated users from nonusers (p = .03). In contrast, 16% of participants self-reported Web use. There was significant discordance between the veracity of actual and self-reported use (p = .004). Among true users, most (81%) logged on once only. These findings raise questions about how to increase use of important health communication interventions.     

Seizure-induced oxidative stress in rat brain regions: blockade by nNOS inhibition

Free radicals have been implicated in the pathogenesis of various neurological disorders including epilepsy. Experimental seizures are often accompanied by the generation of free radicals that cause lipid peroxidation (LPO), which may subsequently cause neurodegeneration observed in certain types of human epilepsy. We recently reported a trigger role for nitric oxide (NO) derived by activation of neuronal isoform of nitric oxide synthase (nNOS) and that the action of conventional antiepileptic drugs (AEDs) was potentiated by inhibition of nNOS. In the present study, we extend our observations to understand the significance of blockade of the nNOS pathway on seizure-induced oxidative stress. Increased NO and LPO levels was observed at the time that corresponded to the onset of generalized seizures in rat brain regions following administration of GABA(A) receptor antagonist, picrotoxin (PCT). Treatment with the selective nNOS inhibitor, 7-nitroindazole (7-NI), decreased NO and LPO levels. The AEDs, diazepam and phenobarbitone also prevented seizure-induced increase in NO and LPO levels. Seizures resulted in a significant increase in the activity of antioxidant enzymes, superoxide dismutase in the frontal cortex and hippocampus. On the other hand, the activity of glutathione peroxidase was decreased in the hippocampus and midbrain. Whereas treatment with 7-NI could minimize the effects of PCT, the AEDs per se did not have any significant impact on the activity of the antioxidant enzymes, though co-treatment with 7-NI and AEDs could significantly decrease seizure-induced alterations in antioxidant enzyme activities. These observations suggest that the AEDs may not have a significant role in modulating the activities of antioxidant enzymes and that their ability to decrease LPO is realized more likely by their ability to prevent free radical formation. In conclusion, the present study demonstrates that NO contributes to LPO observed following seizures induced by PCT. The study also provides evidence for the ability of the AEDs to inhibit seizure-induced increase in LPO levels, the effect being enhanced by co-treatment with 7-NI suggesting that 7-NI and the AEDs together could prevent the neurotoxic cascade induced by oxidative stress.