Unique Reagent Research Articles

Discovery of A Synthetic Hub for Regio- and Stereoselective Construction of Triazolyl Vinyl Sulfonyl Fluorides.

A new sulfonyl fluoride reagent 1-bromobut-3-ene-1,3-disulfonyl difluoride (BEDF) was developed. This unique reagent possesses two clickable functionalities to be used for both azide-alkyne cycloaddition click and SuFEx click reactions. This new reagent was applied for the regioselective construction of a class of novel triazolyl vinyl sulfonyl fluorides in which the C-4 position 1H-1,2,3-triazoles were functionalized with vinyl sulfonyl fluorides of exclusively E-configuration.

Pd/C-Catalyzed Selective N-Monomethylation by Transfer Hydrogenation of Urea Derivatives using Methanol as H2 and C1 Sources.

N-monomethyl amines are useful intermediates in drugs, natural products, paints. Yet their synthesis is a tremendous challenge due to their high reactivity, typically leading to overmethylation. In this contribution, a highly selective catalytic N-methylation methodology is reported, converting urea derivatives to monomethylated amines, using a commercially available heterogeneous Pd/C catalyst and methanol as unique reagent. Methanol provides a sustainable alternative protocol for the selective preparation of mono-methylated derivatives as it acts as both H2 and C1 sources. In addition, several control experiments were performed to provide a proposal for the mechanism, namely dehydrogenation of methanol and subsequent hydrogenation of urea derivatives, followed by reduction of the in situ formed methyl imine. Importantly, the approach is simple, highly productive and enables novel synthetic procedures for the preparation of monomethylamines from urea derivatives.

Chemical reagent for detecting tension wood in selected tree species

Reaction wood is a wood defect arising during the growth of the tree in the part of the trunk that is under tension (hardwood tree species) or compression (coniferous tree species). Beech (Fagus sylvatica L.) tension wood has different anatomical and chemical characteristics than normal (opposite) wood. The difference in density is conditioned by the percentage of the gelatinous layer (G-layer). Fibre cells in reaction beech wood have a different cell wall structure and a different chemical composition. Tension wood cannot be detected by the naked eye. It is only possible to assume its occurrence based on the macroscopic characteristics of the logs, such as a woolly surface, taper or eccentric pith, and so forth. However, these are imprecise and unreliable methods that have minimal effectiveness, especially when shortening the length of the log for cut-outs. This study aimed to create a unique chemical reagent for the detection of tension wood in logs and timber and wood products immediately. The present research can contribute to the mitigation of flaws resulting from the reaction of wood in timber production while addressing noticeable constraints in manufacturing, such as energy resources and the availability of wood raw materials. This can be achieved through the efficient identification of reaction wood in materials. The colour change is only temporary and will fade over time. After the chemical reagent has dried on the surface, the surface can be milled. The colour change extends to a depth of approx. of 3 to 5 mm.

A new method for the robust expression and single-step purification of dCas9 for CRISPR interference/activation (CRISPRi/a) applications

CRISPR-Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats-CRISPR associated enzyme 9) is known for its simplicity, versatility, and scalability in genome editing applications. In vitro Cas9, when complexed with sgRNA, binds and cleaves the complementary target sequences with almost perfect precision. The enzyme is exploited for various applications in understanding and changing gene function. dCas9 (deactivated or dead Cas9) is a double mutated version of Cas9 that bears mutations in the nuclease domains of the enzyme and thus cannot cleave the target DNA. dCas9 is equally advantageous since it can alter gene expression using various transcriptional activators CRISPRa and repressors CRISPRi. Additionally, dCas9 can bind to the desired target gene without cleaving it, making it a unique reagent to study the kinetics and stability of RNA-protein-DNA interactions required to design more efficient and specific gene-editing nucleases. An appreciable quantity of pure and homogeneous protein is needed to characterise dCas9 for its structural and functional understanding. This study used an N-terminal acidic tag to express the dCas9 in an E. coli-bacterial host. A simple single-step protocol for robust and efficient production of dCas9 has been described. The study and methods are distinctive as the purification is performed in a single step using inexpensive multi-modal hydroxyapatite chromatography. The purified protein can be used in different in vitro and in vivo studies.

Redox-Active Boron Clusters.

ConspectusIn this Account, we discuss our group's research over the past decade on a class of functionalized boron clusters with tunable chemical and physical properties, with an emphasis on accessing and controlling their redox behavior. These clusters can be thought of as three-dimensional aromatic systems that have distinct redox behavior and photophysical properties compared to their two-dimensional organic counterparts. Specifically, our lab has studied the highly tunable, multielectron redox behavior of B12(OR)12 clusters and applied these molecules in various settings. We first discuss the spectroscopic and electrochemical characterization of B12(OR)12 clusters in various oxidation states, followed by their use as catholytes and/or anolytes in redox flow batteries and chemical dopants in conjugated polymers. Additionally, the high oxidizing potential and visible light-absorbing nature of fluoroaryl-functionalized B12(OR)12 clusters have been leveraged by our group to generate weakly coordinating, photoexcitable species that can promote photooxidation chemistry.We have further translated these solution-phase studies of B12(OR)12 clusters to the solid state by using the precursor [B12(OH)12]2- cluster as a robust building block for hybrid metal oxide materials. Specifically, we have shown that the boron cluster can act as a thermally stable cross-linking material, which enhances electron transport between metal oxide nanoparticles. We applied this structural motif to create TiO2- and WO3-containing materials that showed promising properties as photocatalysts and electroactive materials for supercapacitors. Building on this concept, we later discovered that B12(OCH3)12, the smallest of the B12(OR)12 family, could retain its redox behavior in the solid state, a previously unseen phenomenon. We successfully harnessed this unique behavior for solid-state energy storage by implementing this boron cluster as a cathode-active material in a Li-ion prototype cell device. Recently, our group has also explored how to tune the redox properties of clusters other than B12(OR)12 species by synthesizing a library of vertex-differentiated clusters containing both B-OR and B-halogen groups. Due to the additive qualities of different functional groups on the cluster, these species allow access to a region of electrochemical potentials previously inaccessible by fully substituted closo-dodecaborate alkoxy-based derivatives.Lastly, we discuss our research into smaller-sized redox-active polyhedral boranes (B6- and B10-based cluster cores). Interestingly, these clusters show significantly less redox stability and reversibility than their dodecaborate-based counterparts. While exploring the functionalization of closo-hexaborate to create fully substituted derivates (i.e., [B6R6Hfac]-), we observed unique oxidative decomposition pathways for this cluster system. Consequently, we leveraged this oxidative instability to generate useful alkyl boronate esters via selective chemical oxidation. We further explored a closo-decaborate cluster as a platform to access electrophilic [B10H13]+ species capable of directly borylating arene compounds with unique regioselectivity. Upon chemical oxidation of the arylated decaborate clusters, we successfully synthesized various aryl boronate esters, establishing the generality of the oxidative cluster deconstruction concept.Overall, our work shows that boron clusters are an appealing class of redox-active molecules, and this fundamental and understudied property can be leveraged for constructing novel materials with tunable physical and electrochemical properties, as well as producing unique chemical reagents for small molecule synthesis.

Trimethylsilanol Cleaves Stable Azaylides As Revealed by Unfolding of Robust "Staudinger" Single-Chain Nanoparticles.

Herein, we disclose a unique and selective reagent for the cleavage of stable azaylides prepared by the nonhydrolysis Staudinger reaction, enabling the on-demand unfolding of robust single-chain nanoparticles (SCNPs). SCNPs with promising use in catalysis, nanomedicine, and sensing are obtained through intrachain folding of discrete synthetic polymer chains. The unfolding of SCNPs involving reversible interactions triggered by a variety of external stimuli (e.g., pH, temperature, light, and redox potential) or substances (e.g., competitive reagents, solvents, and anions) is well known. Conversely, methods for the unfolding (i.e., intrachain disassembly) of SCNPs with stronger covalent interactions are scarce. We show that trimethylsilanol (Me3SiOH) triggers the efficient unfolding of robust "Staudinger" SCNPs with stable azaylide (-N=P-) moieties as intrachain cross-linking units showing exceptional stability toward water, air, and CS2, a standard reagent for azaylides. As a consequence, Me3SiOH arises as a rare, exceptional, and valuable reagent for the cleavage of stable azaylides prepared by the nonhydrolysis Staudinger reaction.

A single domain antibody-based Luminex assay for the detection of SARS-CoV-2 in clinical samples.

Within the past decade, single domain antibodies (sdAbs) have been recognized as unique affinity binding reagents that can be tailored for performance in a variety of immunoassay formats. Luminex MagPlex color-coded magnetic microspheres provide a high-throughput platform that enables multiplexed immunoassays. We developed a MagPlex bead-based assay for the detection of SARS-CoV-2, using sdAbs against SARS-CoV-2 nucleocapsid (N) protein in which we engineered the sdAb capture reagents to orient them on the beads. The oriented sdAbs provided an increase in sensitivity over randomly oriented sdAbs for samples of N diluted in buffer, which also translated into better detection of SARS-CoV-2 in clinical samples. We assessed the specificity of the assay by examining seasonal coronavirus clinical samples. In summary, we provide a proof-of-concept that a bead-based assay using sdAbs to detect SARS-CoV-2 is feasible and future research combining it with other sdAb-coated beads that can detect other viruses may provide a useful diagnostic tool.

Virtual plates: Getting the best out of high content screens

High content screening (HCS) is becoming widely adopted as a high throughput screening modality, using hundred-of-thousands compounds library. The use of machine learning and artificial intelligence in image analysis is amplifying this trend. Another factor is the recognition that diverse cell phenotypes can be associated with changes in biological pathways relevant to disease processes. There are numerous challenges in HCS campaigns. These include limited ability to support replicates, low availability of precious and unique cells or reagents, high number of experimental batches, lengthy preparation of cells for imaging, image acquisition time (45–60 min per plate) and image processing time, deterioration of image quality with time post cell fixation and variability within wells and batches. To take advantage of the data in HCS, cell population based rather than well-based analyses are required. Historically, statistical analysis and hypothesis testing played only a limited role in non-high content high throughput campaigns. Thus, only a limited number of standard statistical criteria for hit selection in HCS have been developed so far. In addition to complex biological content in HCS campaigns, additional variability is impacted by cell and reagent handling and by instruments which may malfunction or perform unevenly. Together these can cause a significant number of wells or plates to fail. Here we describe an automated approach for hit analysis and detection in HCS. Our approach automates HCS hit detection using a methodology that is based on a documented statistical framework. We introduce the Virtual Plate concept in which selected wells from different plates are collated into a new, virtual plate. This allows the rescue and analysis of compound wells which have failed due to technical issues as well as to collect hit wells into one plate, allowing the user easier access to the hit data.

B-315 Investigation Into Extraction Procedure Substitution in the Abbott Architect i2000 Tacrolimus, Sirolimus, and Cyclosporine Assays

Abstract Background Tacrolimus, cyclosporine, and sirolimus are commonly used immunosuppressive drugs with narrow therapeutic windows requiring consistent monitoring to avoid drug toxicity. The Abbott Architect i2000 tacrolimus, sirolimus, and cyclosporine assays each require a pre-analytical whole blood lysing extraction step prior to analysis on the i2000. This extraction is largely manual, requiring the combination of an extraction reagent with the patient sample, followed by an incubation and centrifugation/pelleting steps, after which, the resulting supernatant is assayed for drug concentration. Each assay has its own unique extraction reagent and incubation steps, albeit all three procedures are quite similar. A clinician-requested investigation into a questionably elevated tacrolimus level led us to investigate the effect of substituting extraction reagents and procedures between the three assays. In this study, we investigated the effect of substituting extraction procedures and reagents on assayed immunosuppressive drug values between the Abbott tacrolimus, sirolimus, and cyclosporine immunoassays. Methods Whole blood was obtained from three patients who had previously had either their tacrolimus, sirolimus, or cyclosporine level assayed within the normal therapeutic range. For each patient, the cyclosporine, tacrolimus, or sirolimus level was re-assayed in triplicate, with the substitution of the other two assay’s extraction procedures performed prior to analysis. Such as: tacrolimus extraction was performed, then assayed for sirolimus or cyclosporine. This procedure was repeated for all three drugs and extraction procedures, affording 9 trials in total. The procedure was repeated with three samples obtained from patients with no measurable immunosuppressive drug levels. Additionally, cross-reactivity in the assay extraction reagents was assessed by measuring, in triplicate, immunosuppressive drug concentration on each extraction reagent without the addition of patient whole blood. Results Analysis in patients undergoing immunosuppressive drug therapy afforded the following changes in assayed drug concentration: Tacrolimus substituting sirolimus, cyclosporine extractions, 5.60 to 8.13, 8.47 ng/mL; sirolimus substituting cyclosporine, tacrolimus extractions 13.63 to 4.60, 8.07 ng/mL; cyclosporine substituting sirolimus, tacrolimus extractions 274.6 to 391.3, 757.30 ng/mL. Analysis in patients with no measurable drug content resulted in no change in assayed drug concentration with exception of tacrolimus, in which tacrolimus substituting sirolimus, cyclosporine extractions afforded <1.50 to 4.40, 4.00 ng/mL. Cross-reactivity trials yielded the following drug concentrations: tacrolimus precipitant: tacrolimus: >30.00, cyclosporine: >1500, sirolimus: 5.97 ng/mL; cyclosporine precipitant: cyclosporine: >1500, tacrolimus: 7.13, sirolimus 1.53 ng/mL; sirolimus precipitant: sirolimus: 5.00, tacrolimus: 8.67, cyclosporine: <25.00 ng/mL. Conclusions The use of incorrect extraction reagents and procedures resulted in both increased and decreased assayed immunosuppressive drug concentrations. These changes in drug concentration were observed in both samples containing, and not containing, immunosuppressive drugs. Significant cross-reactivity was observed in all three assay extraction reagents resulting inappropriately elevated assayed drug concentrations in all three assays. This highlights the need for comprehensive technologist training and effective pre-analytical workflows to prevent errors, such as reagent substitution, from occurring. This study has clinical implications into investigations of spurious immunosuppressive drug values, and substitution of assay extraction reagents and procedures should be investigated when performing root cause analysis into incorrectly reported immunosuppressive drug concentrations.

Pooled genetic screens to identify vulnerabilities in TERT-promoter-mutant glioblastoma.

Pooled genetic screens represent a powerful approach to identify vulnerabilities in cancer. Here we used pooled CRISPR/Cas9-based approaches to identify vulnerabilities associated with telomerase reverse transcriptase (TERT) promoter mutations (TPMs) found in >80% of glioblastomas. We first developed a platform to detect perturbations that cause long-term growth defects in a TPM-mutated glioblastoma cell line. However, we could not detect dependencies on either TERT itself or on an E-twenty six transcription (ETS) factor known to activate TPMs. To explore this finding, we cataloged TPM status for 441 cell lines and correlated this with genome-wide screening data. We found that TPM status was not associated with differential dependency on TERT, but that E-twenty six (ETS) transcription factors represent key dependencies in both TPM+ and TPM- lines. Further, we found that TPMs are associated with expression of gene programs regulated by a wide array of ETS-factors in both cell lines and primary glioblastoma tissues. This work contributes a unique TPM cell line reagent, establishes TPM status for many deeply-profiled cell lines, and catalogs TPM-associated vulnerabilities. The results highlight challenges in executing genetic screens to detect TPM-specific vulnerabilities, and suggest redundancy in the genetic network that regulates TPM function with therapeutic implications.

In Memoriam: Soldano Ferrone MD, PhD (1940–2023)

In Memoriam: Soldano Ferrone MD, PhD (1940–2023)

Abstract 6438: Next-generation pan-cancer immunotherapy with patient-derived APC

Abstract Harnessing phagocytic antigen presenting cells (APC) and tumor neoantigen-specific T cells represent the ultimate power of immunotherapy against cancer. The promises of phagocytic APCs lie in their abilities of not only initiating phagocytosis towards cancer cells (to “eat off” tumor) but also, through immunogenic antigen presentation, activating neoantigen-specific Tcells that systemically and durably eliminate cancer cells, as well as the establishment of a long-lasting cellular and hormonal immunity that prevents tumor recurrence. However, the efforts of developing APCs -based therapy have been hindered by the current lack of technology that robustly produce APCs from cancer patients, as none of the traditional methods for differentiating DC or macrophage APCs are effective for cancer patients. Through a proprietary process, we have created a unique reagent combo that enables robust differentiation of cancer patients’ peripheral monocytes (cMo) to be potent APC, termed κAPC. κAPC display a unique gene expression profile, separating from classical DC or macrophage, and profound proinflammatory phenotype with enhanced phagocytic capability and elevated immunogenic antigen presentation machinery. In both in vitro and in vivo settings, κAPC demonstrate direct uptake of tumor antigens and presentation to T cells, the latter leading to activation and large expansion of multi-targeting, polyclonal tumor-specific CD4 and CD8 T populations - termed NeoT- from circulating and tumor-infiltrating lymphocytes. Theexpanded CD8 NeoT are highly tumoricidal and once adoptively transferred in vivo, potently eliminated late-stage metastatic solid tumors, whereas CD4 NeoT contribute to immune memory and repolarize a proinflammatory TME favorable for tumor elimination. Key words: κAPC, NeoT, NeoT adoptive cell therapy, κAPC-based cancer vaccine, combination therapies Citation Format: Zhen Bian, Lei Shi, Yuan Liu, harry Stylli. Next-generation pan-cancer immunotherapy with patient-derived APC [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6438.

Synthesis of spirotriazolines and spirooxadiazolines via light-induced 1,3-dipolar [3+2] cycloadditions

We developed a strategy for the construction of spirotriazolines and spirooxadiazolines by the light-induced [3+2] cycloaddition with 2,5-diaryltetrazole. The use of light as the unique reagent is a distinguishing feature of this protocol.

Trans-Trifluoromethyltetrafluorosulfanyl Chloride: Selective Synthesis and Reaction with Diazo Compounds.

trans-Trifluoromethyltetrafluorosulfanyl chloride (trans-CF3SF4Cl) is a unique reagent for the incorporation of the CF3SF4 group into organic compounds. However, CF3SF4Cl was prepared from hazardous reagents or formed as mixture of trans and cis isomers in low yield. Herein, a silver-promoted selective synthesis of trans-CF3SF4Cl under safe gas-reagent-free conditions is described. Furthermore, the synthetic application of trans-CF3SF4Cl is demonstrated through the new trifluoromethyltetrafluorosulfanylation of α-diazo carbonyl compounds.

CO Effect on the Dynamics of Platinum Nucleation/Growth Under the Liquid-Phase Synthesis of Pt/C Electrocatalysts

This paper shows how bubbling the reaction medium with CO affects the nucleation/growth of Pt nanoparticles under the synthesis using formic acid or formaldehyde as reducing agents. CO has been found to affect not only the dynamics of the Pt(IV) to transformation but also the size of the formed particles (1.1–3.9 nm) deposited on the carbon support (ca. 20 wt% Pt). The dual role of CO due to combining the functions of a stabilizer and a reducing agent has been shown. The study demonstrates that the introduction of the dispersed carbon support to the reaction medium leads to the enhancement of the CO positive effect on the microstructure of Pt/C catalysts. In this case, the spatial distribution of Pt nanoparticles over the carbon support surface becomes more uniform that is conducive to increasing the electrochemical surface area. Thus, CO is a unique reagent used to control the microstructure of Pt nanoparticles during their wet synthesis.

Characterization of KIR+ NK cell subsets with a monoclonal antibody selectively recognizing KIR2DL1 and blocking the specific interaction with HLA-C.

The phenotypic identification of different NK cell subsets allows more in‐depth characterization of KIR repertoire and function, which are of potential interest in KIR and disease association studies. KIR genes are highly polymorphic, but a great homology exists among the various sequences and few monoclonal antibodies (mAbs) specifically recognize a single KIR. This is the case of HP‐DM1 which was demonstrated by analysis of cell transfectants and epitope mapping to be exclusively KIR2DL1‐specific, covering all allotypes identified to date, except for KIR2DL1*022 and *020, and also to react with KIR2DS1*013. Here, we compared in immunofluorescence analyses the staining of HP‐DM1 with other available mAbs to precisely identify KIR2DL1+ NK cells in potential donors for αβT/B‐depleted haplo‐HSCT, with known KIR genotype. HP‐DM1 mAb was used in combination with EB6 or 11PB6 (anti‐KIR2DL1/S1 and anti‐KIR2DL3*005), 143211 (anti‐KIR2DL1/S5), and HP‐MA4 (anti‐KIR2DL1/S1/S3/S5) mAbs, allowing the accurate identification of different KIR+ NK cell subsets. These phenotypic evaluations appeared useful to dissect the expression pattern of various KIR2D in NK cells from KIR2DL3*005 + individuals, particularly if KIR2DS1 is present. HP‐DM1 mAb remarkably refined NK cell phenotyping of donors carrying KIR2DS5, either in the centromeric or telomeric region. Functional assays with KIR2DL1+/S1+/S5+ NK cells confirmed that only HP‐DM1 exclusively reacts with KIR2DL1. Finally, we demonstrated that HP‐DM1 mAb blocked KIR2DL1 recognition of C2+ HLA‐C. Altogether, the data support that HP‐DM1 is a unique reagent valuable for characterizing KIR+ NK cell subsets.

Cu-Catalyzed C(sp3)–N Coupling and Alkene Carboamination Enabled by Ligand-Promoted Selective Hydrazine Transfer to Alkyl Radicals

Selective delivery of an N-nucleophile onto an alkyl radical is a challenging step in redox catalysis. In this work, di-tert-butyl hydrazodiformate was found to be a unique effective amination reagent in Cu-catalyzed C(sp3)–N bond formation reactions that involve alkyl radical intermediates. This method is applicable to both electron-deficient and electron-rich radicals, and we were thus able to achieve the direct C–N coupling of activated bromides, as well as the carboamination of general alkenes based on this chemistry. Mechanistic studies suggest that the hydrazodiformate forms a reducing dinuclear complex with Cu and the ligand which, upon 1e-oxidation, turns into an open-shell species with the major spin density on N atoms. With the assistance of a ligand, this species can selectively deliver the hydrazine moiety onto an alkyl radical (prior to halide transfer), leading to the formation of a C(sp3)–N bond. The products obtained with this amination method could be easily deprotected to afford alkyl hydrazines and further derived to primary amines or N-heterocycles.

Interleukin-6 triggers toxic neuronal iron sequestration in response to pathological α-synuclein.

SUMMARYα-synuclein (α-syn) aggregation and accumulation drive neurodegeneration in Parkinson’s disease (PD). The substantia nigra of patients with PD contains excess iron, yet the underlying mechanism accounting for this iron accumulation is unclear. Here, we show that misfolded α-syn activates microglia, which release interleukin 6 (IL-6). IL-6, via its trans-signaling pathway, induces changes in the neuronal iron transcriptome that promote ferrous iron uptake and decrease cellular iron export via a pathway we term the cellular iron sequestration response, or CISR. The brains of patients with PD exhibit molecular signatures of the IL-6-mediated CISR. Genetic deletion of IL-6, or treatment with the iron chelator deferiprone, reduces pathological α-syn toxicity in a mouse model of sporadic PD. These data suggest that IL-6-induced CISR leads to toxic neuronal iron accumulation, contributing to synuclein-induced neurodegeneration.

Long-term, infection-acquired immunity against the SARS-CoV-2 Delta variant in a hamster model.

The emergence of the SARS-CoV-2 Delta variant (B.1.617.2) raises concerns about potential reduced sensitivity of the virus to antibody neutralization and subsequent vaccine breakthrough infections. Here, we use a live virus neutralization assay with sera from Pfizer- and Moderna-vaccinated individuals to examine neutralizing antibody titers against SARS-CoV-2 and observe a 3.9- and 2.7-fold reduction, respectively, in neutralizing antibody titers against the Delta variant compared with an early isolate bearing only a D614G substitution in its spike protein. We observe similar reduced sensitivity with sera from hamsters that were previously infected with an early isolate of SARS-CoV-2. Despite this reduction in neutralizing antibody titers against the Delta variant, hamsters previously infected (up to 15 months earlier) with an early isolate are protected from infection with the Delta variant, suggesting that the immune response to the first infection is sufficient to provide protection against subsequent infection with the Delta variant.

TMSCF2Br‐Enabled Fluorination–Aminocarbonylation of Aldehydes: Modular Access to α‐Fluoroamides

AbstractA protocol for the modular assembly of the α‐fluoroamide motif has been developed, which provides a practical method for the efficient synthesis of structurally diverse α‐fluoroamides from easily available aldehydes and tertiary amines through a three‐component fluorination–aminocarbonylation process. The key to the success of this process is taking advantage of the multiple roles of the unique difluorocarbene reagent TMSCF2Br (TMS=trimethylsilyl). The mechanism of the process involves the 1,2‐fluorine and oxygen migrations of the in situ formed TMS‐protected α‐aminodifluoromethyl carbinol intermediates, which represents a new type of deoxyfluorination reaction.