Process Mass Intensity (PMI): A Holistic Analysis of Current Peptide Manufacturing Processes Informs Sustainability in Peptide Synthesis.

Small molecule therapeutics represent the majority of the FDA-approved drugs. Yet, many attractive targets are poorly tractable by small molecules, generating a need for new therapeutic modalities. Due to their biocompatibility profile and structural versatility, peptide-based therapeutics are a possible solution. Additionally, in the past two decades, advances in peptide design, delivery, formulation, and devices have occurred, making therapeutic peptides an attractive modality. However, peptide manufacturing is often limited to solid-phase peptide synthesis (SPPS), liquid phase peptide synthesis (LPPS), and to a lesser extent hybrid SPPS/LPPS, with SPPS emerging as a predominant platform technology for peptide synthesis. SPPS involves the use of excess solvents and reagents which negatively impact the environment, thus highlighting the need for newer technologies to reduce the environmental footprint. Herein, fourteen American Chemical Society Green Chemistry Institute Pharmaceutical Roundtable (ACS GCIPR) member companies with peptide-based therapeutics in their portfolio have compiled Process Mass Intensity (PMI) metrics to help inform the sustainability efforts in peptide synthesis. This includes PMI assessment on 40 synthetic peptide processes at various development stages in pharma, classified according to the development phase. This is the most comprehensive assessment of synthetic peptide environmental metrics to date. The synthetic peptide manufacturing process was divided into stages (synthesis, purification, isolation) to determine their respective PMI. On average, solid-phase peptide synthesis (SPPS) (PMI ≈ 13,000) does not compare favorably with other modalities such as small molecules (PMI median 168-308) and biopharmaceuticals (PMI ≈ 8300). Thus, the high PMI for peptide synthesis warrants more environmentally friendly processes in peptide manufacturing.

Unseen overlap between fishing vessels and top predators in the northeast Pacific.

Accurate assessments of human-wildlife risk associated with industrial fishing are critical for the conservation of marine top predators. Automatic Identification System (AIS) data provide a means of mapping fishing and estimating human-wildlife risk; however, risk can be obscured by gaps in the AIS record due to technical issues and intentional disabling. We assessed the extent to which unseen fishing vessel activity due to AIS gaps obscured estimates of overlap between fishing vessel activity and 14 marine predators including sharks, tunas, mammals, seabirds, and critically endangered leatherback turtles. Among vessels equipped with AIS in the northeast Pacific, up to 24% of total predator overlap with fishing vessel activity was unseen, and up to 36% was unseen for some individual species. Waters near 10°N had high unseen overlap with sharks yet low reported shark catch, revealing potential discrepancies in self-reported datasets. Accounting for unseen fishing vessel activity illuminates hidden human-wildlife risk, demonstrating challenges and solutions for transparent and sustainable marine fisheries.

The History of Flow Chemistry at Eli Lilly and Company.

Flow chemistry was initially used for speed to early phase material delivery in the development laboratories, scaling up chemical transformations that we would not or could not scale up batch for safety reasons. Some early examples included a Newman Kwart Rearrangement, Claisen rearrangement, hydroformylation, and thermal imidazole cyclization. Next, flow chemistry was used to enable safe scale up of hazardous chemistries to manufacturing plants. Examples included high pressure hydrogenation, aerobic oxidation, and Grignard formation reactions. More recently, flow chemistry was used in Small Volume Continuous (SVC) processes, where highly potent oncolytic molecules were produced by fully continuous processes at about 10 kg/day including reaction, extraction, distillation, and crystallization, using disposable equipment contained in fume hoods.

Dynamic interplay: disentangling the temporal variability of fish effects on coral recruitment.

Ecosystems around the world are continuously undergoing recovery from anthropogenic disturbances like climate change, overexploitation, and habitat destruction. Coral reefs are a prime example of a threatened ecosystem and coral recruitment is a critical component of reef recovery from disturbances. Reef fishes structure this recruitment by directly consuming macroalgae and coral recruits or by indirectly altering the substrate to facilitate coral settlement (e.g., grazing scars). However, how these direct and indirect mechanisms vary through time remains largely unknown. Here, we quantified coral recruitment on settlement tiles with divots that mimic grazing scars and caging treatments to exclude or allow fish feeding over 3 years at Palmyra Atoll in the Pacific Ocean. We found that the positive and negative effects of fishes on coral recruitment varies through time. After 3 years, both grazing scars and fish grazing no longer predicted coral recruitment, suggesting that the role of fishes decreases over time. Our results emphasize that reef fish populations are important in promoting initial coral recovery after disturbances. However, over time, factors like the environment may become more important. Future work should continue to explore how the strength and direction of top-down control by consumers varies through time across multiple ecosystems.

Consumers decrease variability across space and turnover through time during coral reef succession.

Consumers play an integral role in mediating ecological succession-the change in community composition over time. As consumer populations are facing rapid decline in ecosystems around the world, understanding of their ecological role is becoming increasingly urgent. Increased understanding of how changes in consumer populations may influence community variability across space and turnover through time during succession is particularly important for coral reefs, which are among the most threatened ecosystems globally, and where fishes play vital roles in structuring benthic succession. Here, we examine how consumers influence coral reef succession by deploying 180 paired settlement tiles, caged (to exclude fishes larger than approximately 15 cm) and uncaged, within Palmyra Atoll, a remote marine wildlife refuge with previously documented high fish abundance, and monitored benthic community development one and three years after deployment. We found that excluding large fishes lead to lower alpha diversity and divergent community states across space (i.e.,, high beta diversity among caged tiles), suggesting that benthic fish feeding maintains local diversity but tends to homogenize community composition with dominance by crustose coralline algae. In addition, when fish were experimentally excluded, the developing benthic community exhibited a greater change in species composition over time (i.e., high temporal beta diversity), indicating that fish feeding tends to canalize community successional trajectories. Finally, the caged and uncaged tiles became more similar over time, suggesting that fish feeding plays a more important role during early succession. Our results demonstrate that the loss of large fishes, for example from overfishing, may result in benthic communities that are more variable across space and time. Increased variability could have important implications for ecosystem function and coral reef resilience in the face of escalating global stressors.

Differences in the behavior and diet between shoaling and solitary surgeonfish (Acanthurus triostegus).

Variation in behavior within marine and terrestrial species can influence the functioning of the ecosystems they inhabit. However, the contribution of social behavior to ecosystem function remains underexplored. Many coral reef fish species provide potentially insightful models for exploring how social behavior shapes ecological function because they exhibit radical intraspecific variation in sociality within a shared habitat. Here, we provide an empirical exploration on how the ecological function of a shoaling surgeonfish (Acanthurus triostegus) may differ from that of solitary conspecifics on two Pacific coral reefs combining insight from behavioral observations, stable isotope analysis, and macronutrient analysis of gut and fecal matter. We detected important differences in how the social mode of A. triostegus affected its spatial and feeding ecology, as well as that of other reef fish species. Specifically, we found increased distance traveled and area covered by shoaling fish relative to solitary A. triostegus. Additionally, shoaling A. triostegus primarily grazed within territories of other herbivorous fish and had piscivorous and nonpiscivorous heterospecific fish associated with the shoal, while solitary A. triostegus grazed largely grazed outside of any territories and did not have any such interactions with heterospecific fish. Results from stable isotope analysis show a difference in δ15N isotopes between shoaling and solitary fish, which suggests that these different social modes are persistent. Further, we found a strong interaction between social behavior and site and carbohydrate and protein percentages in the macronutrient analysis, indicating that these differences in sociality are associated with measurable differences in both the feeding ecology and nutrient excretion patterns. Our study suggests that the social behavior of individuals may play an important and underappreciated role in mediating their ecological function.

Mechanistic Study of Diketopiperazine Formation during Solid-Phase Peptide Synthesis of Tirzepatide.

This study focused on investigating diketopiperazine (DKP) and the formation of associated double-amino-acid deletion impurities during linear solid-phase peptide synthesis (SPPS) of tirzepatide (TZP). We identified that the DKP formation primarily occurred during the Fmoc-deprotection reaction and post-coupling aging of the unstable Fmoc-Pro-Pro-Ser-resin active pharmaceutical ingredient (API) intermediate. Similar phenomena have also been observed for other TZP active pharmaceutical ingredient (API) intermediates that contain a penultimate proline amino acid, such as Fmoc-Ala-Pro-Pro-Pro-Ser-resin, Fmoc-Pro-Pro-Pro-Ser-resin, and Fmoc-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-resin, which are intermediates for both hybrid and linear synthesis approaches. During post-coupling aging, it is found that Fmoc deprotection can proceed in dimethylformamide (DMF), dimethyl sulfoxide (DMSO), N-methyl-2-pyrrolidone (NMP), and acetonitrile (ACN) solvents without any piperidine addition. Density functional theory (DFT) calculations showed that a peptide that has a penultimate proline stabilizes the transition state through the C-H···π interaction during Fmoc decomposition, which causes those peptides to be more prone to cascade-deprotection reactions. Pseudo-reaction pathways are then proposed, and a corresponding macrokinetics model is developed to allow accurate prediction of the TZP peptide intermediate self-deprotection and DKP formation rate. Based on those studies, control strategies for minimizing DKP formation were further investigated and an alternative to Fmoc protection was identified (Bsmoc-protected amino acids), which eliminated the formation of the DKP byproducts. In addition, the use of oxyma additives and lower storage temperature was demonstrated to markedly improve the peptide intermediate stability to DKP degradation pathways.

Hot spots of unseen fishing vessels.

Illegal, unreported, and unregulated (IUU) fishing incurs an annual cost of up to US$25 billion in economic losses, results in substantial losses of aquatic life, and has been linked to human rights violations. Vessel tracking data from the automatic identification system (AIS) are powerful tools for combating IUU, yet AIS transponders can be disabled, reducing its efficacy as a surveillance tool. We present a global dataset of AIS disabling in commercial fisheries, which obscures up to 6% (>4.9 M hours) of vessel activity. Disabling hot spots were located near the exclusive economic zones (EEZs) of Argentina and West African nations and in the Northwest Pacific, all regions of IUU concern. Disabling was highest near transshipment hot spots and near EEZ boundaries, particularly contested ones. We also found links between disabling and location hiding from competitors and pirates. These inferences on where and why activities are obscured provide valuable information to improve fisheries management.

Tracking the response of industrial fishing fleets to large marine protected areas in the Pacific Ocean.

Large marine protected areas (MPAs) of unprecedented size have recently been established across the global oceans, yet their ability to meet conservation objectives is debated. Key areas of debate include uncertainty over nations' abilities to enforce fishing bans across vast, remote regions and the intensity of human impacts before and after MPA implementation. We used a recently developed vessel tracking data set (produced using Automatic Identification System detections) to quantify the response of industrial fishing fleets to 5 of the largest MPAs established in the Pacific Ocean since 2013. After their implementation, all 5 MPAs successfully kept industrial fishing effort exceptionally low. Detected fishing effort was already low in 4 of the 5 large MPAs prior to MPA implementation, particularly relative to nearby regions that did not receive formal protection. Our results suggest that these large MPAs may present major conservation opportunities in relatively intact ecosystems with low immediate impact to industrial fisheries, but the large MPAs we considered often did not significantly reduce fishing effort because baseline fishing was typically low. It is yet to be determined how large MPAs may shape global ocean conservation in the future if the footprint of human influence continues to expand. Continued improvement in understanding of how large MPAs interact with industrial fisheries is a crucial step toward defining their role in global ocean management.

Seguimiento a la Respuesta de las Flotillas de Pesca Industrial a las Grandes Áreas Marinas Protegidas Extensas Resumen Recientemente se han establecido grandes áreas marinas protegidas (AMPs) de tamaños nunca vistos en todos los océanos del mundo; sin embargo, se sigue debatiendo su habilidad para lograr los objetivos de conservación. El debate se centra en los siguientes temas importantes: la incertidumbre por la capacidad de las naciones para hacer cumplir las vedas de pesca en regiones vastas y remotas y la intensidad del impacto humano antes y después de la implementación de una AMP. Usamos un conjunto de datos de rastreo de navíos recientemente desarrollado (producido usando detecciones mediante el Sistema Automático de Identificación) para cuantificar la respuesta de las flotillas de pesca industrial ante cinco de las AMPs más grandes establecidas en el océano Pacífico desde 2013. Después de su implementación, las cinco AMPs mantuvieron exitosamente los esfuerzos de pesca industrial a niveles excepcionalmente bajos. El esfuerzo de pesca detectado ya se encontraba bajo en cuatro de las cinco grandes AMPs previo a la implementación, particularmente en relación con las regiones próximas que no reciben protección formal. Nuestros resultados sugieren que estas grandes AMPs pueden presentar oportunidades importantes de conservación en ecosistemas relativamente intactos con un impacto inmediato bajo para las pesquerías industriales, pero las grandes AMPs que consideramos con frecuencia no redujeron significativamente el esfuerzo de pesca porque la línea base de la pesca con frecuencia ya era baja. Todavía se debe determinar cómo las grandes AMPs pueden moldear la conservación mundial de los océanos en el futuro si la huella de la influencia humana continúa expandiéndose. La mejoría continua del entendimiento de cómo las grandes AMPs interactúan con las pesquerías industriales es un paso importante hacia la definición de su papel en el manejo mundial de los océanos.

Predicted hotspots of overlap between highly migratory fishes and industrial fishing fleets in the northeast Pacific.

Many species of sharks and some tunas are threatened by overexploitation, yet the degree of overlap between industrial fisheries and pelagic fishes remains poorly understood. Using satellite tracks from 933 industrial fishing vessels and predictive habitat models from 876 electronic tags deployed on seven shark and tuna species, we developed fishing effort maps across the northeast Pacific Ocean and assessed overlap with core habitats of pelagic fishes. Up to 35% of species' core habitats overlapped with fishing effort. We identified overlap hotspots along the North American shelf, the equatorial Pacific, and the subtropical gyre. Results indicate where species require international conservation efforts and effective management within national waters. Only five national fleets (Mexico, Taiwan, China, Japan, and the United States) account for >90% of overlap with core habitats of our focal sharks and tunas on the high seas. These results inform global negotiations to achieve sustainability on the high seas.

Response to Comment on "Tracking the global footprint of fisheries".

Amoroso et al demonstrate the power of our data by estimating the high-resolution trawling footprint on seafloor habitat. Yet we argue that a coarser grid is required to understand full ecosystem impacts. Vessel tracking data allow us to estimate the footprint of human activities across a variety of scales, and the proper scale depends on the specific impact being investigated.

Tracking the global footprint of fisheries.

Although fishing is one of the most widespread activities by which humans harvest natural resources, its global footprint is poorly understood and has never been directly quantified. We processed 22 billion automatic identification system messages and tracked >70,000 industrial fishing vessels from 2012 to 2016, creating a global dynamic footprint of fishing effort with spatial and temporal resolution two to three orders of magnitude higher than for previous data sets. Our data show that industrial fishing occurs in >55% of ocean area and has a spatial extent more than four times that of agriculture. We find that global patterns of fishing have surprisingly low sensitivity to short-term economic and environmental variation and a strong response to cultural and political events such as holidays and closures.

Kilogram-scale prexasertib monolactate monohydrate synthesis under continuous-flow CGMP conditions.

Advances in drug potency and tailored therapeutics are promoting pharmaceutical manufacturing to transition from a traditional batch paradigm to more flexible continuous processing. Here we report the development of a multistep continuous-flow CGMP (current good manufacturing practices) process that produced 24 kilograms of prexasertib monolactate monohydrate suitable for use in human clinical trials. Eight continuous unit operations were conducted to produce the target at roughly 3 kilograms per day using small continuous reactors, extractors, evaporators, crystallizers, and filters in laboratory fume hoods. Success was enabled by advances in chemistry, engineering, analytical science, process modeling, and equipment design. Substantial technical and business drivers were identified, which merited the continuous process. The continuous process afforded improved performance and safety relative to batch processes and also improved containment of a highly potent compound.

Mild and direct conversion of quinoline N-oxides to 2-amidoquinolines with primary amides.

[reaction: see text] A simple, one-pot procedure is described for the direct conversion of quinoline N-oxides to alpha-amidoquinolines with primary amides. This methodology is complimentary to the Abramovich reaction, which is limited to the introduction of secondary amides via imidoyl chlorides. Although reaction conditions are quite similar, omission of the base is key for successful reaction with primary amides, which were found not to proceed through the intermediacy of an imidoyl chloride but rather through an acyl isocyanate.

Stereoselective Nazarov cyclizations of bridged bicyclic dienones.

[reaction: see text] Bridged bicyclic dienones underwent silyl-directed Nazarov cyclization with generally very high diastereoselectivity. In most cases, a strong preference for cyclization to the product with an exo-disposed cyclopentenone was seen. However, the presence of additional unsaturation in the three-carbon bridge of a bicyclo[3.2.1]octadiene system caused complete reversal in selectivity with exclusive formation of the endo-cyclopentenone. The observed selectivities are believed to result from a combination of steric and electronic effects.