OSMI-4
目录号 : GC31517
OSMI-4 是迄今为止报道的最好的 OGT 抑制剂，OSMI-4 的活性形式具有低纳摩尔结合亲和力。
Cas No.:2260791-14-6
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >99.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
| Kinase experiment [1]: | |
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Preparation Method |
OGT reactions were carried out in a 50-µL final volume, containing 0.1 mM UDP-GlcNAc, 200 nM purified full-length OGT, OSMI-4a/b,100 µM RBL-2 peptide in OGT reaction buffer. Reactions were incubated at 37℃ for 2 h. Afterwards, each reaction was transferred in duplicate into a 96-well white microplate and was mixed with a 1:1 ratio of the UDP-Glo Detection Reagent. After incubation at room temperature for 1 h, the luminescence was recorded with a POLARstar® Omega microplate reader or with a BioTek Synergy™ H4 microplate reader. |
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Reaction Conditions |
37℃ for 2 h |
|
Applications |
IC50 of OSMI-4a and OMSI-4b are 1.5 and 0.5μM, respectively. |
| Cell experiment [2]: | |
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Cell lines |
HEK293T |
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Preparation Method |
Cells were plated and grown in their respective media until they reached a confluency of 60-80%. For HEK293T cells, media was changed 3 hours before OSMI-4 was added; OSMI-4 was added directly to each well at the indicated concentrations. The number of dead cells were assessed by CellToxTM Green cytotoxicity Assay. |
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Reaction Conditions |
0.125,0.25,0.5,1,2,4,8,16μM，24 h |
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Applications |
EC50 of OSMI-4 in cells is 3 μM |
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References: [1]. Loi EM, Weiss M, Pajk S, Gobec M, Tomašič T, Pieters RJ, Anderluh M. Intracellular Hydrolysis of Small-Molecule O-Linked N-Acetylglucosamine Transferase Inhibitors Differs among Cells and Is Not Required for Its Inhibition. Molecules. 2020 Jul 25;25(15):3381. [2]. Martin SES, Tan ZW, Itkonen HM, Duveau DY, Paulo JA, Janetzko J, Boutz PL, Törk L, Moss FA, Thomas CJ, Gygi SP, Lazarus MB, Walker S. Structure-Based Evolution of Low Nanomolar O-GlcNAc Transferase Inhibitors. J Am Chem Soc. 2018 Oct 24;140(42):13542-13545. |
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OSMI-4 is the best OGT inhibitor reported to date and the active form of OSMI-4 has low nanomolar binding affinity [1]. OSMI-4 has a chlorine substituent on the quinolinone ortho to the sulfonamide, thus allowing the molecule to bind tighter in the uridine binding pocket within the enzyme’s active site [2]. The free acid and ester forms of OSMI-4 are OSMI-4a and OSMI-4b, respectively.
OSMI-4 has further enhanced binding affinities to the enzyme with a Kd of 8 nM. The IC50 of OSMI-4a and OMSI-4b are 1.5 and 0.5μM, respectively [3]. The EC50 of OSMI-4 in HEK293T cells is 3 μM [1].
References:
[1].Martin SES, Tan ZW, Itkonen HM, Duveau DY, Paulo JA, Janetzko J, Boutz PL, T?rk L, Moss FA, Thomas CJ, Gygi SP, Lazarus MB, Walker S. Structure-Based Evolution of Low Nanomolar O-GlcNAc Transferase Inhibitors. J Am Chem Soc. 2018 Oct 24;140(42):13542-13545.
[2].Ju Kim E. O-GlcNAc Transferase: Structural Characteristics, Catalytic Mechanism and Small-Molecule Inhibitors. Chembiochem. 2020 Nov 2;21(21):3026-3035.
[3].Loi EM, Weiss M, Pajk S, Gobec M, Toma?i? T, Pieters RJ, Anderluh M. Intracellular Hydrolysis of Small-Molecule O-Linked N-Acetylglucosamine Transferase Inhibitors Differs among Cells and Is Not Required for Its Inhibition. Molecules. 2020 Jul 25;25(15):3381.
OSMI-4 是迄今为止报道的最好的 OGT 抑制剂，OSMI-4 的活性形式具有低纳摩尔结合亲和力[1]。 OSMI-4 在与磺酰胺邻位的喹啉酮上有一个氯取代基，因此允许该分子在酶活性位点 [2] 内的尿苷结合袋中更紧密地结合。 OSMI-4的游离酸和酯形式分别为OSMI-4a和OSMI-4b。
OSMI-4 进一步增强了对该酶的结合亲和力，Kd 为 8 nM。 OSMI-4a和OMSI-4b的IC50分别为1.5和0.5μM[3]。 OSMI-4在HEK293T细胞中的EC50为3 μM [1]。
| Cas No. | 2260791-14-6 | SDF | |
| Canonical SMILES | O=C1C=CC2=C(C=C(Cl)C(S(N[C@@H](C(N(CC(OCC)=O)CC3=CC=CS3)=O)C4=C(OC)C=CC=C4)(=O)=O)=C2)N1 | ||
| 分子式 | C27H26ClN3O7S2 | 分子量 | 604.09 |
| 溶解度 | DMSO : 125 mg/mL (206.92 mM) | 储存条件 | Store at -20°C |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.6554 mL | 8.2769 mL | 16.5538 mL |
| 5 mM | 0.3311 mL | 1.6554 mL | 3.3108 mL |
| 10 mM | 0.1655 mL | 0.8277 mL | 1.6554 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Design of OSMI-4 analogs using scaffold hopping: investigating the importance of the uridine mimic in the binding of OGT inhibitors
β-N-Acetylglucosamine transferase (OGT) inhibition is considered an important topic in medicinal chemistry. The involvement of O-GlcNAcylation in several important biological pathways is pointing to OGT as a potential therapeutic target. The field of OGT inhibitors drastically changed after the discovery of the 7-quinolone-4-carboxamides scaffold and its optimization to the first nanomolar OGT inhibitor: OSMI-4. While OSMI-4 is still the most potent inhibitor reported to date, its physicochemical properties are limiting its use as a potential drug candidate as well as a biological tool. In this study, we have introduced a simple modification (elongation) of the peptide part of OSMI-4 that limits the unwanted cyclisation during OSMI-4 synthesis while retaining OGT inhibitory potency. Secondly, we have kept this modified peptide unchanged while incorporating new sulfonamide UDP mimic to try to improve binding of newly designed OGT inhibitors in the UDP-binding site. With the use of computational methods, a small library of OSMI-4 derivatives was designed, prepared and evaluated that provided information about the OGT binding pocket and its specificity toward quinolone-4-carboxamides.
Release of O-GlcNAc transferase inhibitor promotes neuronal differentiation of neural stem cells in 3D bioprinted supramolecular hydrogel scaffold for spinal cord injury repair
Precise fabrication of biomimetic three-dimensional (3D) structure and effective neuronal differentiation under the pathological environment are the key to neural stem cell (NSC)-based spinal cord injury (SCI) therapy. In this study, we have developed a spinal cord-like bioprinted scaffold loading with OSMI-4, a small molecule O-GlcNAc transferase (OGT) inhibitor, to induce and guide the neuron differentiation of NSCs for efficient SCI repair. To achieve this, we developed a supramolecular bioink (SM bioink) consisting of methacrylated gelatin and acrylated β-cyclodextrins to load NSCs and OSMI-4. This bioink showed fast gelation and stable mechanical properties, facilitating bioprinting of functional neural scaffolds. Moreover, the weak host-guest cross-linking of the SM scaffolds significantly improved the cell-matrix interaction for the infiltration and migration of NSCs. What's more, the sustained delivery of OSMI-4 remarkably enhanced the intrinsic neuronal differentiation of the encapsulated NSCs in vitro by inhibiting Notch signaling pathway. In vivo experiment further revealed that the functional bioprinted scaffolds promoted the neuronal regeneration and axonal growth, leading to significant locomotor recovery of the SCI model rats. Together, the NSC-laden bioprinted SM scaffolds in combination with sustained release of the therapeutic agent OSMI-4 largely induced neuronal differentiation of NSCs and thus leading to efficient SCI repair. STATEMENT OF SIGNIFICANCE: Efficient neuronal differentiation of neural stem cells (NSCs) under the complex pathological microenvironment of spinal cord injury (SCI) is a major challenge of neural regeneration. By the use of a supramolecular bioink, we bioprinted a spinal cord-like scaffold loaded with NSCs and a small molecule drug OSMI-4 to significantly induce neuronal differentiation of NSCs for efficient SCI repair in vivo. The scaffolds with spinal cord-like structure can support the interaction and neuronal differentiation of NSCs by providing a dynamic matrix and a source of molecular release of OSMI-4. The influences of OSMI-4 on NSCs and its molecular mechanism were investigated for the first time in this study. Altogether, three-dimensional bioprinting fabrication of NSC- and small molecule drug-laden biomimetic construct may represent a promising therapeutic strategy for SCI repair.
Intracellular Hydrolysis of Small-Molecule O-Linked N-Acetylglucosamine Transferase Inhibitors Differs among Cells and Is Not Required for Its Inhibition
O-GlcNAcylation is an essential post-translational modification that occurs on nuclear and cytoplasmic proteins, regulating their function in response to cellular stress and altered nutrient availability. O-GlcNAc transferase (OGT) is the enzyme that catalyzes this reaction and represents a potential therapeutic target, whose biological role is still not fully understood. To support this research field, a series of cell-permeable, low-nanomolar OGT inhibitors were recently reported. In this study, we resynthesized the most potent OGT inhibitor of the library, OSMI-4, and we used it to investigate OGT inhibition in different human cell lines. The compound features an ethyl ester moiety that is supposed to be cleaved by carboxylesterases to generate its active metabolite. Our LC-HRMS analysis of the cell lysates shows that this is not always the case and that, even in the cell lines where hydrolysis does not occur, OGT activity is inhibited.