Development of ß-cyclodextrin-based sustained release microparticles for oral insulin delivery.

Polymeric microparticles have been previously demonstrated to deliver various therapeutic agents efficiently to targeted regions by protecting the drug from harsh gastric milieu of the gastrointestinal tract. In this study, we investigated the hypoglycemic effect of ß-cyclodextrin polymeric insulin microparticles in diabetic rats via the oral route of administration. ß-cyclodextrin microparticles were prepared by a unique one-step spray-drying technique and stabilized by incorporating enteric retardant polymers in the formulation. The insulin-loaded microparticles had a mean size of 0.8 ± 0.25 µm with a zeta potential of 3.57 + 0.62 mV. As seen with the chromatographic analysis, the drug content in the microparticles was determined to be 94.9 ± 2.77%. RAW macrophage cells showed greater than 80% viability after 24 h of incubation with the insulin and blank microparticles. For the in vitro release study, the microparticles were able to protect the insulin in gastric fluid where no significant release was detected, followed by only 50% release in intestinal fluid for the first 8 h of the study. This was seen to correlate with the in vivo data where 50% glucose inhibition was seen after 8 h of oral administration in diabetic rats. This data suggest that the oral insulin microparticles were able to reduce glucose levels in disease conditions and would be a favorable route of administration to patients as an alternative to daily subcutaneous injections.

Ascertaining and promoting effective study skills and learning habits of first-year pharmacy students.

INTRODUCTION: Many pharmacy students struggle academically in their initial professional year. This research ascertained first-year pharmacy (P1) students' study skills and learning habits both before and after employing interventions about effective study strategies. METHODS: In 2018-2019 and 2019-2020, P1 students completed a study skills inventory at baseline in August and at endpoint in April. Interventions included instruction about the science of learning and effective study techniques as well as the use of a cognitive wrapper regarding their first assessment. RESULTS: Students' perceived habits that improved were those of organizing their ideas, studying at their peak time, and really wanting to learn the content. Perceived habits that declined included time spent doing their assigned readings, seeing the need to always attend class, and reviewing course material each day. Regarding the cognitive wrapper, 90% of students thought they were prepared for their assessment, yet 21% received a lower grade than expected and 10% did not work on practice problems at least 2 days before the quiz. CONCLUSIONS: This endeavor elucidated several important areas about the study skills and learning habits of P1 students. Some students' perceptions of their study skills and learning habits strengthened and others decreased during their first year. The cognitive wrapper provided a targeted way for students to reflect on their preparation and performance as well as consider their future study plans. Faculty can use this information to help students employ effective learning practices to promote students' metacognition throughout the first year of their program.

A dual-delivery platform for vaccination using antigen-loaded nanoparticles in dissolving microneedles.

Effective vaccines delivered via painless methods would revolutionize the way people approach vaccinations. This study focused on the development of fast-dissolving microneedles (MNs) to deliver antigen-loaded sustained release polymeric nanoparticles (NPs), achieving a dual-delivery platform for vaccination through the skin. The platform utilizes dissolving MNs (dMNs), which penetrate to the epidermal layer of the skin and rapidly dissolve, releasing the antigen-loaded NPs. In this study, seven dissolving microneedle formulations were tested based on screening of various biocompatible and biodegradable polymers and sugars. The lead dMN formulation was selected based on optimal mechanical strength and dissolution of the needles and was loaded with poly(lactic-co-glycolic) acid (PLGA) NPs encapsulating a model influenza matrix 2 (M2) protein antigen. Antigen-loading efficiency in the needles was determined by centrifugation of the lead formulation containing various concentrations of antigen nanoparticles. Next, the reproducibility and translatability of ex vivo mechanical strength and dissolvability of the lead M2 PLGA NP-loaded dMN formulation was assessed by formulating and testing two different microneedle arrays on murine and porcine skin. Finally, the lead microneedle array was loaded with fluorescent dye NPs and evaluated for pore formation and closure in vivo in a murine model. This proof-of-concept study yielded an easy-to-formulate, well-characterized, translatable antigen NP-loaded dMN platform for transdermal vaccine administration.

Crafting Immunological Response Using Particulate Vaccines.

To achieve optimal immunogenicity, particulates present a promising vehicle for antigen delivery and have the potential to skew immune response. Particulate vaccine offers several advantages including targeting of antigen to sentinel cells, protection from degradation, sustained release, and itself acts an adjuvant mimics viral structure. Adjuvant presence is vital in overcoming the poor immunogenicity of vaccines, e.g., subunit vaccines. Adjuvants have antigen dose sparing potential and provide danger signals to alert the immune system. Various particulate carriers received attention in the delivery of vaccine antigens such as virus-like particles, liposomes, immunostimulating complexes, and polymeric particles. This review also discussed the properties of particles such as size, shape, and rigidity affecting the immunological outcome. It further highlights the cellular uptake of the particulate vaccine, antigen processing, and its presentation by antigen-presenting cells. For mass vaccination, especially in countries lacking resources, effect of storage temperature condition on stability of vaccine is pivotal. The current COVID-19 pandemic is not showing any signs of abatement and role of nanocarriers are highly relevant in SARS-CoV-2 pandemic as an effective immunization strategy. Eradication of pandemic demands the rapid evaluation of multiple approaches that can provides successful vaccination platform, enabling scalability and global distribution.

A novel microparticulate vaccine for melanoma cancer using transdermal delivery.

In this study, we formulated a microparticulate melanoma cancer vaccine via the transdermal route. The vaccine was delivered using microneedle-based Dermaroller® which is available for cosmetic purposes. Unlike subcutaneous injections, administration using microneedles is painless and in general can increase the permeability of many compounds ranging in size from small molecules to proteins and microparticles that do not normally penetrate the skin. The vaccine microparticles were taken up by the antigen presenting cells which demonstrated a strong IgG titre level of 930 ug/mL in serum samples. The formulation increased the immunogenicity of the vaccine by incorporating the antigen into an albumin matrix having a size range of around 0.63-1.4 µm which acted as a synthetic adjuvant. The animals were vaccinated with 1 prime and 4 booster doses administered every 14 days over 8 weeks duration, followed by challenge with live tumour cells which showed protection after transdermal vaccination.

Multicenter, randomized, double-blind, active comparator and placebo-controlled trial of a corticotropin-releasing factor receptor-1 antagonist in generalized anxiety disorder.

BACKGROUND: Antagonism of corticotropin-releasing factor (CRF) receptors has been hypothesized as a potential target for the development of novel anxiolytics. This study was designed to determine the safety and efficacy of pexacerfont, a selective CRF-1 receptor antagonist, in the treatment of generalized anxiety disorder (GAD). METHOD: This was a multicenter, randomized, double-blind, placebo-controlled and active comparator trial. Two hundred and sixty patients were randomly assigned to pexacerfont 100 mg/day (after a 1 week loading dose of 300 mg/day), placebo or escitalopram 20 mg/day in a 2:2:1 ratio. The primary outcome was the mean change from baseline to end point (week 8) in the Hamilton Anxiety Scale total score. RESULTS: Pexacerfont 100 mg/day did not separate from placebo on the primary outcome measure. The half-powered active comparator arm, escitalopram 20 mg/day, demonstrated efficacy with significant separation from placebo at weeks 1, 2, 3, 6, and 8 (P<.02). Response rates for pexacerfont, placebo, and escitalopram were 42, 42, and 53%, respectively. Genetic and psychometric rating scale data was obtained in 175 randomized subjects. There was a significant association between a single nucleotide polymorphism (SNP) of the gene encoding plexin A2 (PLXNA2-2016) with the HAM-A psychic subscale score for the entire cohort at baseline (FDR-adjusted P=.015). CONCLUSIONS: Pexacerfont did not demonstrate efficacy compared to placebo for the treatment of GAD. Whether these findings are generalizable to this class of agents remains to be determined. Our preliminary genetic finding of an association between a SNP for the gene encoding plexin A2 and an anxiety phenotype in this study merits further exploration. The trial was registered at clinicaltrials.gov (NCT00481325) before enrollment.

Efficacy and tolerability of flexibly-dosed adjunct TC-5214 (dexmecamylamine) in patients with major depressive disorder and inadequate response to prior antidepressant.

This paper reports the efficacy and tolerability of the nicotinic channel modulator TC-5214 (dexmecamylamine) as adjunct therapy for patients with major depressive disorder who have an inadequate response to initial antidepressant treatment in 2 Phase III studies. These double-blind, placebo-controlled studies (NCT01157078, D4130C00002 [Study 002] conducted in the US and India; NCT01180400, D4130C00003 [Study 003] conducted in Europe) comprised 8 weeks of open-label antidepressant treatment followed by 8 weeks of active treatment during which patients were randomized to flexibly-dosed TC-5214 1-4 mg twice daily (BID) or placebo as an adjunct to ongoing therapy with SSRI/SNRI. The primary efficacy endpoint in both studies was change in Montgomery Åsberg Depression Rating Scale (MADRS) total score from randomization (week 8) to treatment end (week 16). Secondary endpoints included change in Sheehan Disability Scale and Hamilton Depression Rating Scale 17-item scores. Study 002 randomized 319 patients and Study 003 randomized 295 patients to TC-5214 or placebo. At treatment end, no significant differences were seen for change in MADRS total score with TC-5214 versus placebo. Furthermore, there were no significant differences in any of the secondary endpoints. The most commonly reported (≥ 10%) adverse events with TC-5214 in these studies were constipation and headache. In these 2 flexibly-dosed studies, no specific therapeutic effects were observed for TC-5214 (1-4 mg BID) adjunct to antidepressant in the primary endpoint or any secondary endpoint; however, TC-5214 was generally well tolerated. In conclusion, no antidepressant effect of TC-5214 was observed in these studies.

Enhanced bioavailability of orally administered antisense oligonucleotide to nuclear factor kappa B mRNA after microencapsulation with albumin.

Antisense molecules that pertain to ribonucleic acid (RNA) and complementary to the messenger RNA (mRNA) are produced by transcription of a given gene. Antisense oligonucleotides have emerged as potential gene-specific therapeutic agents that are currently undergoing evaluation in clinical trials for a variety of diseases. When administered orally, antisense oligionucleotides have poor bioavailability as they are rapidly degraded by the acid in the stomach and by the enzymes in the intestine. Therefore, the enhancement of bioavailability after oral administration is highly desirable. This article shows the enhanced bioavailability of antisense oligonucleotides that targets nuclear factor kappa B (NF-κB) mRNA after encapsulating in an inert, biodegradable albumin polymer matrix that was administered via the oral route into a rat model. The bioavailability of the antisense oligonucleotides to NF-κB in microencapsulated form was compared to the solution form of the drug upon oral administration. The solution form had a low bioavailability of 9%, whereas the bioavailability for the microencapsulated form of the drug increased up to 70%. Moreover, the other pharmacokinetic parameters including half-life (t1/2) and volume of distribution (Vd) increased for the microencapsulated form compared to the solution form of the drug.

Oral delivery of microparticles containing plasmid DNA encoding hepatitis-B surface antigen.

The role of albumin-based chitosan microparticles on enhancing immune response of plasmid DNA (pDNA) to hepatitis-B surface antigen (HBsAg) vaccine after oral administration was investigated in mice. The pDNA encoding HBsAg was entrapped in albumin microparticles using a one-step spray drying technique optimized in our laboratory. The encapsulated particles were also characterized in vitro for their shape, size, encapsulation efficiency, content, and stability. Albumin microparticles could protect the DNA from nuclease degradation as confirmed in our agarose gel study. Further immune modulating effect was studied in our formulation by measuring IgG antibodies in serum as well as IgA antibodies in fecal extracts. The mice were immunized with a prime dose of 100 µg of pDNA in microparticle formulations with and without interleukins biweekly until week 7 followed by a booster dose of equivalent strength on week 33 to compare the response with the subcutaneous group. The oral immunization with the pDNA to HBsAg microparticles gave significantly higher titer level of both sIgA and IgG at week 9 and 34, respectively, in oral vaccine with interleukins group when compared with the subcutaneous group. Thus, we observed an augmentation of both humoral and cellular immune responses for prolonged periods after immunization.

Oral microparticulate vaccine for melanoma using M-cell targeting.

Cancer vaccines are limited in their use, because of their inability to mount a robust anti-tumor immune response. Thus, targeting M-cells in the small intestine, which are responsible for entry of many pathogens, will be an attractive way to elicit a strong immune response toward particulate antigens. Therefore, in the present investigation, we demonstrated that efficient oral vaccination against melanoma antigens could be accomplished by incorporating the antigens in an albumin-based microparticle with a ligand AAL (Aleuria aurantia lectin) targeted specifically to M-cells. The oral microparticulate vaccine effectively protected the mice from subcutaneous challenge with tumor cells in prophylactic settings. The animals were vaccinated with antigen microparticles having a size range of around 1-1.25 µm where one prime and four booster doses were administered every 14 days over 10 weeks of duration, followed by challenge with live tumor cells, which showed complete tumor protection after oral vaccination. With the inclusion of ligand in the microparticles, we observed significantly higher IgG titers (1565 µg/mL) as compared to the microparticle formulations without AAL (872 µg/mL). This data suggests that ligand loaded microparticles may have the potential to target antigens to M-cells for an efficient oral vaccination.