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Chemical Structure| 3483-12-3 Chemical Structure| 3483-12-3
Chemical Structure| 3483-12-3

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DTT is a redox reagent known as Cleland's reagent and is as a reducing or "deprotecting" agent for thiolated DNA.

Synonyms: DL-dithiothreitol; rel-(2R,3R)-1,4-Dimercapto-2,3-butanediol

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Product Citations

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Shen, Chenfangfei ; Tamminga, Micah J ; Gillies, Elizabeth R ;

Abstract: There is growing interest in self-immolative polymers (SIPs), which can depolymerize end-to-end in response to backbone or end-cap cleavage. The ability of these polymers to respond to specific stimuli has garnered interest for a wide range of applications, including drug and nucleic acid delivery, sensing, micro- and nanoscale patterning, adhesives, and recyclable plastics. Among the promising candidates for SIPs are polythioesters (PTEs), which feature dynamic thioester bonds in their backbone. Despite recent advancements, investigation into the depolymerization behavior of PTEs in aqueous environments remains limited. Herein, we explore the behavior of two different PTE backbones. PTEs based on D-penicillamine and Boc-L-hydroxyproline were synthesized with disulfide and control end-caps. The penicillamine-based polymers depolymerized in response to dithiothreitol (DTT) regardless of their end-cap, indicating that DTT could cleave the polymer backbone, which could be followed by depolymerization and additional backbone cleavage events. On the other hand, the dominant mechanism for the hydroxyproline-based polythioesters appeared to be end-cap cleavage, followed by depolymerization. Overall, both classes of PTEs were responsive to thiols, indicating their potential utility in biomedical conditions involving hypoxia, and thus elevated levels of thiols such as glutathione. As proof of concept, we also demonstrated that the PTEs could be used to prepare degradable gels and self-assembled nanoparticles.

Keywords: Polythioester ; Self-immolative ; Depolymerization ; Reduction-responsive

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Jin, Sebin ; Oh, Dongyeop ; Park, Jeyoung ;

Abstract: The excessive and persistent use of petroleum-derived plastics has resulted in severe environmental challenges due to their long-term durability and resistance to degradation. Herein, we present sustainable PUU elastomers that integrate in-use mechanical robustness and autonomous self-healing, and on-demand multi-pathway degradability. At the core of this system is 4-aminophenyl disulfide, an aromatic disulfide building block that imparts multiple functionalities: its rigid conjugated structure enhances thermal and mechanical stability; its dynamic disulfide bonds undergo rapid exchange at mild temperatures, enabling efficient self-healing; and its primary amine forms urea linkages with diisocyanates. Unlike conventional polyurethanes relying solely on urethane linkages, the introduction of urea bonds with dual hydrogen donors forms stronger, more directional supramolecular networks, enhancing cohesion and toughness. Additionally, ester-rich bio-based polyols enable enzymatic hydrolysis, while disulfide bonds impart redox- and photo-responsive degradability. This molecular architecture effectively breaks the conventional trade-offs between mechanical durability, degradability, and dynamic functionality. This elastomer demonstrates high tensile strength (36 MPa), elongation at break over 400%, and self-healing efficiency up to 98% at 80 °C. It also exhibits excellent UV-induced self-healing, achieving over 90% healing efficiency at ambient temperature after 24 h, while exhibiting reliable degradation under enzymatic, photochemical, and reductive environments.

Keywords: Self-healing ; Poly(urethane-urea) ; Disulfide bond ; Multi-pathway degradation ; Bio-based macro-diol

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Kumpati, Greeshma P ; Ronayne, Conor T ; Johnson, Joseph L ; Gardner, Zachary S ; Singh, Anuj K ; Iyer, Ananth , et al.

Abstract: Glutathione transferase (GSTP1 and GSTM2) are tractable targets for anticancer drug development. In this work, a series of 6-(7-nitro-2,1,3-benzoxadiazol-4-ylthio)hexanol (NBDHEX) based analogues were designed, synthesized, and evaluated both theoretically and experimentally as GSTP1 and GSTM2 inhibitors. Among the synthesized compounds, 3h showed selective inhibition toward GSTP1 while 5b showed selective inhibition of GSTM2. Compounds 5b and 5c exhibited stronger potency while compound 3h showed slightly lower potency against the tested cancer cells than its parent molecule NBDHEX. Comprehensive biological studies were conducted on the effect of 3h, 5b and 5c towards breast cancer MDA-MB-231 and pancreatic MiaPaCa-2 cell lines revealed that 3h, 5b and 5c could activate JNK pathway and induce cell apoptosis. Furthermore in vivo experiments using NSG mice demonstrated that 5b significantly reduced tumor growth when administered in combination with gemcitabine, effectively overcoming gemcitabine resistance in the MiaPaCa-2 cell model through targeted inhibition of GSTM2. These findings suggests that, 5b could become a promising candidate for further development as a potential antitumor agent in cancer therapy.

Keywords: anticancer agents ; oxidative stress ; GSTP1 ; GSTM2 ; JNK ; gemcitabine resistance

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Kaustav Khatua ; Yugendar R. Alugubelli ; Kai S. Yang ; Veerabhadra R. Vulupala ; Lauren R. Blankenship ; Demonta Coleman , et al.

Abstract: The main (MPro) of SARS-CoV-2, the causative agent of COVID-19, is a pivotal nonstructural critical for viral replication and pathogenesis. Its function relies on three active site pockets for substrate recognition and a catalytic for enzymatic activity. To develop potential SARS-CoV-2 , we successfully synthesized a diverse range of azapeptide inhibitors with various covalent warheads to target MPro's catalytic . Our characterization identified potent MPro inhibitors, including MPI89 that features an aza-2,2-dichloroacetyl warhead with a remarkable EC50 value of 10 nM against SARS-CoV-2 infection in ACE2+ A549 cells and a selective index of 875. MPI89 is also remarkably selective and shows no potency against SARS-CoV-2 papain-like and several human proteases. Crystallography analyses demonstrated that these inhibitors covalently engaged the catalytic and used the aza-amide carbonyl oxygen to bind to the oxyanion hole. MPI89 stands as one of the most potent MPro inhibitors, suggesting the potential for further exploration of azapeptides and the aza-2,2-dichloroacetyl warhead for developing effective therapeutics against COVID-19.

Keywords: COVID-19 ; SARS-CoV-2 ; Main protease ; Azapeptide ; Covalent inhibitor

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Yingjie Wu ; Samuel J. Rozans ; Abolfazl Salehi Moghaddam ; E. Thomas Pashuck ;

Abstract: Cells dynamically modify their local extracellular matrix by expressing proteases that degrade matrix proteins. This enables cells to spread and migrate within tissues, and this process is often mimicked in hydrogels through the incorporation of peptide crosslinks that can be degraded by cell-secreted proteases. However, the cleavage of hydrogel crosslinks will also reduce the local matrix mechanical properties, and most crosslinking peptides, such as the widely used GPQGIWGQ “PanMMP” sequence, lead to bulk degradation of the hydrogel. A subset of proteases are localized to the cell membrane and are only active in the pericellular region in the immediate vicinity of the cell surface. These membrane-type proteases have important physiological roles and enable cells to migrate within tissues. In this work we developed an approach to identify and optimize peptide sequences that are specifically degraded by membrane-type proteases. We utilized a proteomic screen to identify peptide targets, and coupled this with a functional assay that both quantifies peptide degradation by individual cell types and can elucidate whether the peptides are primarily cleaved by soluble proteases or membrane-type proteases. We then used a split-and-pool synthesis approach to generate more than 300 variants of the target peptide to improve the degradation behavior. We identified an optimized peptide sequence, KLVADLMASAE, which is primarily degraded by membrane-type proteases, but enables both endothelial cells and stem cells grown in KLVADLMASAE-crosslinked hydrogels to spread and have viabilities similar to the gels crosslinked by the PanMMP peptide. Notably, the biological performance of the KLVADLMASAE peptide-cross linked gels was significantly improved from the initial peptide target found in the proteomic screen. This work introduces a functional approach to identifying and refining protease-substrate peptides as a way to enhance the properties of hydrogel matrices.

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Samuel J. Rozans ; Abolfazl Salehi Moghaddam ; Yingjie Wu ; Kayleigh Atanasoff ; Liliana Nino ; Katelyn Dunne , et al.

Abstract: Peptides are widely used within biomaterials to improve cell adhesion, incorporate bioactive ligands, and enable cell-mediated degradation of the matrix. While many of the peptides incorporated into biomaterials are intended to be present throughout the life of the material, their stability is not typically quantified during culture. In this work we designed a series of peptide libraries containing four different N-terminal peptide functionalizations and three C-terminal functionalization to better understand how simple modifications can be used to reduce non-specific degradation of peptides. We tested these libraries with three cell types commonly used in biomaterials research, including mesenchymal stem/stromal cells (hMSCs), endothelial cells, and macrophages, and quantified how these cell types non-specifically degraded peptide as a function of terminal amino acid and chemistry. We found that peptides in solution which contained N-terminal amines were almost entirely degraded by 48 hours, irrespective of the terminal amino acid, and that degradation occurred even at high peptide concentrations. Peptides with C-terminal carboxylic acids also had significant degradation when cultured with cells. We found that simple modifications to the termini could significantly reduce or completely abolish non-specific degradation when soluble peptides were added to cells cultured on tissue culture plastic or within hydrogel matrices, and that functionalizations which mimicked peptide conjugations to hydrogel matrices significantly slowed non-specific degradation. We also found that there were minimal differences across cell donors, and that sequences mimicking different peptides commonly-used to functionalized biomaterials all had significant non-specific degradation. Finally, we saw that there was a positive trend between RGD stability and hMSC spreading within hydrogels, indicating that improving the stability of peptides within biomaterial matrices may improve the performance of engineered matrices.

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Rozans, Samuel J ; Moghaddam, Abolfazl S ; Pashuck, E Thomas ;

Abstract: Peptides are widely used in biomaterials due to their easy of synthesis, ability to signal cells, and modify the properties of biomaterials. A key benefit of using peptides is that they are natural substrates for cell-secreted enzymes, which creates the possibility of utilizing cell-secreted enzymes for tuning cell-material interactions. However, these enzymes can also induce unwanted degradation of bioactive peptides in biomaterials, or in peptide therapies. Liquid chromatography-mass spectrometry (LC-MS) is a widely used, powerful methodology that can separate complex mixtures of molecules and quantify numerous analytes within a single run. There are several challenges in using LC-MS for the multiplexed quantification of cell-induced peptide degradation, including the need for non-degradable internal standards and the identification of optimal sample storage conditions. Another problem is that cell culture media and biological samples typically contain both proteins and lipids that can accumulate on chromatography columns and degrade their performance. However, removing these constituents can be expensive, time consuming, and increases sample variability. Here we show that directly injecting samples onto the LC-MS without any purification enables rapid and accurate quantification of peptide concentration, and that hundreds of LC-MS runs can be done on a single column without a significantly diminish the ability to quantify the degradation of peptide libraries. We also show that column failure is evident when hydrophilic peptides fail to be retained on the column, and this can be easily identified using standard peptide mixtures for column benchmarking. In total, this work introduces a simple and effective method for simultaneously quantifying the degradation of dozens of peptides in cell culture. By providing a streamlined and cost-effective method for the direct quantification of peptide degradation in complex biological samples, this work enables more efficient assessment of peptide stability and functionality, facilitating the development of advanced biomaterials and peptide-based therapies.

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Dana Qiang Murphy Soika ;

Abstract: Contrast agents (CAs) are small molecules used in magnetic resonance imaging (MRI) to help diagnose various forms of cancer. While MRI is advantageous over other clinical imaging techniques, limitations of today’s contrast agents containing gadolinium (Gd) hinder their safety, sensitivity, and specificity. The conventional CAs that MRI relies on are considered low-relaxivity and are not optimally effective at enhancing MR signal. Additionally, they lack cell-specificity and circulate throughout the body. Furthermore, unbound Gd3+ is nephrogenic which prevents its use in patients with impaired renal function. In the clinic, these limitations mean high dosages of these compounds must be administered to patients in order to produce an image that struggles to highlight the exact tumor location. Our aim was to improve conventional CAs by synthesizing a high-relaxivity (HR) targeted contrast agent (HR-TCA). The cell-specific nature of the HR-TCA will allow for its accumulation at tumor sites while the HR will produce a stronger MR signal per molecule of CA. Combined, this means a much lower and therefore safer dose of CA can be used to produce an image of the exact tumor location with superior contrast. Our modular approach allows us to easily combine this HR contrast agent (HR-CA) to any targeting peptide using a linker in a convergent, one-step synthesis. Our synthetic approach for the HR-CA module attaches a macrocyclic chelator, DO3A, to the side chain of an orthogonally protected alanine. This is a modification to the approach published by Boros in which t-butyl groups were utilized to protect DO3A. In our modular approach, Gd is chelated early to protect the acetic acid donor arms of DO3A from participating in unwanted side reactions for the remainder of the synthesis, eliminating any need to expose the final HR-TCA to the harsh acidic conditions of TFA that are necessary to remove t-butyl protecting groups. Upon removal of the N-and C-terminal protecting groups, the HR-CA module is coupled directly to our in-house synthesized targeting module which is comprised of the targeting agent (DCL) and linker (DSS) already attached to afford the final HR-TCA. T1 relaxation measurements of relevant intermediates and the final product were performed to compare their relaxivities with those of commercial CAs used in clinics, labs, and hospitals today. Although the HR-CA and final HR-TCA exhibited only a modest increase in T1 relaxivity compared to commercial CA Gd-DOTA, in a striking discovery it was observed that the presence of both a tryptophan spacer and an Fmoc protecting group boosted the T1 relaxivity significantly.

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Product Details of DTT

CAS No. :3483-12-3
Formula : C4H10O2S2
M.W : 154.25
SMILES Code : O[C@@H]([C@H](O)CS)CS
Synonyms :
DL-dithiothreitol; rel-(2R,3R)-1,4-Dimercapto-2,3-butanediol
MDL No. :MFCD00004877

Safety of DTT

GHS Pictogram:
Signal Word:Danger
Hazard Statements:H302-H318-H411
Precautionary Statements:P264-P270-P280-P305+P351+P338-P310-P330-P403-P501
Class:9
UN#:3077
Packing Group:
 

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