Quality by Design I: Application of Failure Mode Effect Analysis (FMEA) and Plackett–Burman Design of Experiments in the Identification of “Main Factors” in the Formulation and Process Design Space for Roller-Compacted Ciprofloxacin Hydrochloride Immediate-Release Tablets

As outlined in the ICH Q8(R2) guidance, identifying the critical quality attributes (CQA) is a crucial part of dosage form development; however, the number of possible formulation and processing factors that could influence the manufacturing of a pharmaceutical dosage form is enormous obviating formal study of all possible parameters and their interactions. Thus, the objective of this study is to examine how quality risk management can be used to prioritize the number of experiments needed to identify the CQA, while still maintaining an acceptable product risk profile. To conduct the study, immediate-release ciprofloxacin tablets manufactured via roller compaction were used as a prototype system. Granules were manufactured using an Alexanderwerk WP120 roller compactor and tablets were compressed on a Stokes B2 tablet press. In the early stages of development, prior knowledge was systematically incorporated into the risk assessment using failure mode and effect analysis (FMEA). The factors identified using FMEA were then followed by a quantitative assessed using a Plackett–Burman screening design. Results show that by using prior experience, literature data, and preformulation data the number of experiments could be reduced to an acceptable level, and the use of FMEA and screening designs such as the Plackett Burman can rationally guide the process of reducing the number experiments to a manageable level.KEY WORDS: failure mode effect analysis (FMEA), Plackett–Burman, quality by design (QbD), quality risk management, roller compaction, tablet and ciprofloxacin     

Synthesis of Fmoc-Gly-Ile Phosphinic Pseudodipeptide: Residue Specific Conditions for Construction of Matrix Metalloproteinase Inhibitor Building Blocks

The efficient synthesis of an Fmoc-Gly-Ile phosphinic pseudodipeptide was desired as an eventual building block for construction of matrix metalloproteinase inhibitors. A Michael-type addition reaction of bis(trimethylsilyl) phosphonite with the appropriate acrylate generated the pseudodipeptide bond. Additional of adamantyl (Ad) protection by our prior route (reaction of in situ generated phosphinic acid chloride with the sodium salt of adamantanol) was surprisingly inefficient. Adamantyl protection was achieved in high yield by refluxing the phosphinic acid, Ag₂O, and 1-AdBr in chloroform. Subsequently a concise one-pot three-step reaction comprising a double deprotection of the N- and C-termini under catalytic hydrogenation conditions followed by selective protection of the N-terminus with an Fmoc group yielded Fmoc-NHCH₂PO(OAd)CH₂CH(2-butyl)CO₂H in 41?% overall yield. These results indicate that, as the diversity of phosphinic pseudodipeptides is increased to create selective matrix metalloproteinase inhibitors, different synthetic pathways may be required for efficient building block preparation.     

Sensitive Windows of Skeletal Development in Rabbits Determined by Hydroxyurea Exposure at Different Times throughout Gestation

The critical periods of axial skeletal development in rats and mice have been well characterized, however the timing of skeletal development in rabbits is not as well known. It is important to have a more precise understanding of this timing of axial skeletal development in rabbits due to the common use of this species in standard nonclinical studies to assess embryo–fetal developmental toxicity. Hydroxyurea, a teratogen known to induce a variety of fetal skeletal malformations, was administered to New Zealand White rabbits as a single dose (500 mg/kg) on individual days during gestation (gestation day,GD 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 19) and fetal external, visceral, and skeletal morphology was examined following cesarean sections on GD 29. A wide range of fetal skeletal effects was observed following hydroxyurea treatment, with a progression of malformations from anterior to posterior structures over time, as well as from proximal to distal structures over time. The sensitive window of axial skeletal development was determined to be GD 8 to 13, while disruption of appendicular and cranio‐facial skeletal development occurred primarily from GD 11 to 16 and GD 11 to 12, respectively. The results of this study provide a better understanding of the critical developmental window for different segments of the rabbit skeleton, which will aid in the design of window studies to investigate teratogenicity in rabbits.     

Alternative plants for development of picture‐winged fly pests of maize

Eleven species of picture‐winged flies (Diptera: Ulidiidae: Lipsanini) have been reported attacking maize [Zea mays L. (Poaceae)] ears in the Americas. Four of these species are sweet corn pests in America north of Mexico: Chaetopsis massyla (Walker), Euxesta annonae (Fabricius), E. eluta Loew, and E. stigmatias Loew. Adults of these four species appear at the beginning of each season following maize‐free periods, suggesting other plants act as food sources for maintenance and development of these flies. Studies were conducted in Florida, USA, to evaluate the suitability of several crop and non‐crop plants commonly occurring near maize plantings as developmental hosts for these flies. Laboratory trials were conducted using laboratory colonies of C. massyla, E. eluta, and E. stigmatias to determine their developmental rates and pupal survivorship on roots, stems, leaves, or fruits of 14 crop and weed species. All three fly species completed development on all tested crops (Brassica oleracea L., Capsicum chinense Jacquin, Capsicum annum L., Carica papaya L., Persea americana Mill., Raphanus sativus L., Saccharum officinarum L., and Solanum lycopersicum L.) and weed species [Amaranthus spinosus L., Portulaca oleracea L., Sorghum halepense (L.), and Typha spp.], except for Daucus carota L. roots and Solanum tuberosum L. tubers. Findings of the current study suggest that the presence of multiple host crops in areas surrounding maize fields may help explain the occurrence of these maize‐infesting ulidiids immediately after prolonged absences of maize.     

Candidate chemoreceptor subfamilies differentially expressed in the chemosensory organs of the molluscAplysia

Background  

Marine molluscs, as is the case with most aquatic animals, rely heavily on olfactory cues for survival. In the molluscAplysia californica, mate-attraction is mediated by a blend of water-borne protein pheromones that are detected by sensory structures called rhinophores. The expression of G protein and phospholipase C signaling molecules in this organ is consistent with chemosensory detection being via a G-protein-coupled signaling mechanism.     

Microsatellite markers for the silver arowana (Osteoglossum bicirrhosum, Osteoglossidae, Osteoglossiformes)

Osteoglossum bicirrhosum (silver arowana) is an important fish for the economy of the Amazon region, both as an ornamental fish and as a food fish. To provide tools for addressing ecological and genetic questions, we developed 19 polymorphic microsatellite markers that had between 2 and 7 alleles per locus in the 24 tested individuals. The transferability of many of the loci was confirmed for Osteoglossum ferreirai (black arowana) and Arapaima gigas, and for three African osteoglossiform species.     

Microsatellite markers for the tambaqui (Colossoma macropomum, Serrasalmidae, Characiformes), an economically important keystone species of the Amazon River floodplain

Colossoma macropomum is a keystone species of the Amazon floodplain, and is an important but severely overexploited commercial species. To provide tools for addressing ecological and conservation questions, we developed 14 highly polymorphic microsatellite markers that had between four and 21 alleles per locus in the 25 tested individuals. With the exception of comparisons involving the locus Cm1F5 that also showed heterozygosity deficiency, no pairs of loci were at linkage disequilibrium. Many of the microsatellite loci were also variable in three other serrasalmid species which span the phylogenetic depth of the Serrasalmidae.     

Microsatellite loci for population and parentage analysis in the Amazon River dolphin (Inia geoffrensis de Blainville, 1817)

We developed specific primers for microsatellite DNA regions for the Amazon River dolphin or boto Inia geoffrensis, for use in population and conservation genetic studies. We also tested their transferability for two other species, Pontoporia blainvillei (sister taxon of I. geoffrensis) and Sotalia guianensis. A total of 12 microsatellite loci were polymorphic for the boto. An additional 25 microsatellite loci previously isolated from other cetacean species were also tested in the boto. The 26 polymorphic microsatellite loci indicate they will be excellent markers for studies of population structure and kinship relations of the boto.     

Isolation and characterization of 12 dinucletiotide microsatellite loci in Paratrechalea galianoae (Araneae, Trechaleidae), a nuptial gift-spider

Twelve microsatellite loci from the Neotropical spider Paratrechalea galianoae were isolated from a microsatellite-enriched genomic library, and were tested in a sample of 27 individuals from a population that occurs in Brazil. All loci were polymorphic and three to 14 (mean 7.58) alleles per locus were identified. The observed and expected heterozygosities per locus ranged from 0.417 to 0.885 and from 0.595 to 0.878, respectively. Two loci deviated significantly from Hardy-Weinberg equilibrium and one pair showed evidence of linkage disequilibrium. These loci are being used to study the population structure and genetic diversity in P. galianoae.