Recilisib |CAS 334969-03-8|DC Chemicals

Cas No.:	334969-03-8
Chemical Name:	Benzoic acid,4-[(1E)-2-[[(4-chlorophenyl)methyl]sulfonyl]ethenyl]-
Synonyms:	Benzoic acid,4-[(1E)-2-[[(4-chlorophenyl)methyl]sulfonyl]ethenyl]-;4-[(1E)-2-[[(4-CHLOROPHENYL)METHYL]SULFONYL]ETHENYL]-BENZOIC ACID;ON01210;ON-01210;Recilisib
SMILES:	O=C(O)C1=CC=C(/C=C/S(=O)(CC2=CC=C(Cl)C=C2)=O)C=C1
Formula:	C₁₆H₁₃O₄Scl
M.Wt:	336.79002
Purity:	>98%
Sotrage:	2 years -20°C Powder, 2 weeks 4°C in DMSO, 6 months -80°C in DMSO

Description:	Recilisib (ON01210, EX-RAD) is a radioprotectant that activates the activity of AKT and PI3K in cells. It has been studied as prophylactic (use prior to radiation exposure) and therapeutic (after exposure to radiation) drug.
Target:	Akt PI3K
In Vivo:	Recilisib Sodium (500 mg/kg) significantly increases the rate of recovery and differentiation of primitive bone marrow myeloid progenitor cells in mice. Recilisib Sodium in combination with radiation reduces CFU numbers in mice, but the Recilisib Sodium-treated mice consistently retain a capability to form differentiated colonies. Recilisib Sodium treated mice have a progenitor cell population that is never completely depleted by radiation exposure[1].
In Vitro:	Recilisib Sodium (up to 50 µM) shows a normal distribution of cells throughout the cell cycle, with a slight reduction in the number of cells in S-phase at 50 µM. Continuous exposure of Recilisib Sodium (100 µM) does not result in cell death. Recilisib Sodium does not inhibit the ability of human bone marrow to form colonies in methylcellulose at either timepoint. Recilisib Sodium treatment does not inhibit the colony forming potential of human bone marrow cells. Recilisib Sodium provides dose dependent protection of human bone marrow cells at all three doses of IR. Recilisib Sodium activates the phosphorylation of AKT and GSK3α/β in HFL cells. Recilisib Sodium increases PI3K activity in HFL-1 cells and murine bone marrow cells in response to radiation exposure. Recilisib Sodium treatment in combination with radiation alters the MAPK signaling pathway[1].
Kinase Assay:	PI3-kinase assays are performed using exponentially growing HFL-1 or freshly harvested murine bone marrow cells that are treated with increasing concentrations of Recilisib Sodium for 2 hours and then irradiated with 10 Gy IR. These cells are then returned to the incubator for 2 to 24 hours and lysed in HEPES pH 7.5 lysis buffer. PI-3K is immunoprecipitated using an anti-PI3 Kinas polyclonal antibody for 2 hours at 4°C. Protein A/G PLUS-Agarose is incubated with immunoprecipitates for 8-16 hours at 4°C and the resulting immunoprecipitates washed with twice HEPES pH 7.5 lysis buffer and once with the kinase buffer (20 mM Tris pH 7.5, 1mM EGTA, 10mM MgCl2, 2 mM DTT, 0.01% NP-40). L-α-Phosphatidylinositol (12.5 mM) and ATP (10 µM) are added to the kinase buffer (60 µL per sample) and incubated at 30°C for 30 minutes. The reaction is stopped by addition of 100 µL of 1N HCl and extracted by addition of 200 µL CHCl3/CH3OH (1:1). The extracted samples are vortexed, centrifuged and the lower organic phases containing phospholipids are dried at 27°C for 2 hours. The dried samples are resuspended in 10 µL of PI-4-P standard (0.5 mL CHCl3, 0.5 mL CH3OH, 2.5 µL HCl) and spotted on TLC plates (VWR). The spotted plate is subjected to thin layer chromatography in CHCl3/CH3OH/NH4OH (40:40:15). The TLC plate is dried and subjected to autoradiography.
Cell Assay:	For cytotoxicity assays, cells (1.0×105 cells/mL HPGM) are treated with various concentrations of Recilisib Sodium or vehicle without radiation treatment for 2 or 24 hours. The cells are washed and plated into methocult using gridded dishes. The total number of colony forming units (CFUs) is determined 14 days post-plating by microscopic observation using an Olympus IMT-2 microscope.
References:	[1]. Kang AD, et al. ON01210.Na (Ex-RAD) mitigates radiation damage through activation of the AKT pathway. PLoS One. 2013;8(3):e58355.

MSDS

Cat. No.	Product name	Field of application
DC12145	DLinDMA	DLinDMA is a key lipid component of stable nucleic acid lipid particles as a benchmark.
DC40169	DMG-PEG2000	DMG-PEG 2000 is used for the preparation of liposome for siRNA delivery with improved transfection efficiency in vitro. DMG-PEG 2000 is also used for the lipid nanoparticle for an oral plasmid DNA delivery approach in vivo through a facile surface modific
DC45611	DMPC	1,2-Dimyristoyl-sn-glycero-3-phosphocholine (DMPC) is a synthetic phospholipid used in liposomes. 1,2-Dimyristoyl-sn-glycero-3-phosphocholine is used for the study of lipid monolayers and bilayers.
DC70000	Lysyllysyllysine	Lysyllysyllysine is a cationic moietie that may be used in the construction of gene delivery vectors and DNA nanoparticles.
DC33580	DODMA	DODMA, also known as MBN 305A is a a cationic lipid containing the unsaturated long-chain (18:1) oleic acid inserted at both the sn-1 and sn-2 positions. It has been used in the composition of lipospomes formulated as stable nucleic acid lipid particles that can encapsulate siRNA or other small molecules to be used for drug delivery
DC33635	DODAP	DODAP, also known as 1,2-Dioleoyl-3-dimethylammonium-propane, is a cationic lipid. It has been used as a component in liposomes that can be used to encapsulate siRNA, immunostimulatory oligodeoxynucleotides, antisense oligonucleotides, or chemotherapeutic agents for in vitro and in vivo delivery.
DC59010	C14-4 (C14-494,Lipid B-4,Lipid B4)	C14-4 (C14-494,Lipid B-4,Lipid B4) is a novel ionizable lipid with the highest T-cell transfection efficiency and low cytotoxicity.The C14-4 ionizable lipid has been explored for CAR-T therapy.To screen the excellent formulations for mRNA delivery, a lipid library of 24 ionizable lipids was constructed to make iLNPs, which were used to deliver luciferase mRNA into Jurkat cells.[115] The optimal iLNPs formulation was C14-4 iLNPs (C14-4 ionizable lipid, DOPE, chol, and PEG at a molar ratio of 35%, 16%, 46.5%, and 2.5%) (Figure 6c). The optimal dose of luciferase mRNA for C14-4 iLNPs was 30 ng. Compared with electroporated CAR T cells, the CAR T cells engineered via C14-4 iLNPs showed potent cancer-killing activity when they were cocultured with Nalm-6 acute lymphoblastic leukemia cells. To obtain a safer and more effective CAR mRNA delivery vehicle, the orthogonal design provided 256 potential formulations, and 16 representative iLNPs formulations were evaluated.Through evaluating the safety, delivery efficiency, and transfection efficiency of 16 iLNPs, the formulation B10 (C14-4 ionizable lipid, DOPE, chol, PEG at a molar ratio of 40%, 30%, 25%, and 2.5%) was screened out as the optimal performing formulation. The luciferase expression based on B10 formulation was increased threefold than the initial formulation. Reducing the accumulation and clearance of iLNPs in the liver can increase the expression of CAR mRNA in T cells, further improving the therapeutic effect of CAR-T. Studies have shown that cholesterol analogs can alter the mechanisms of intracellular circulation and enhance the delivery of mRNA, which may be related to the reduced recognition of iLNPs by the Niemann Pick C1 (NPC1) enzyme.The addition of a hydroxyl group to various locations in the cholesterol molecule can alter the binding kinetics between the modified cholesterol and NPC1, and reduced NPC1 recognition of cholesterol. The results showed that replacement of 25% and 50% 7 α-hydroxycholesterol for cholesterol in iLNPs improved mRNA delivery to primary human T cells in vitro by 1.8-fold and twofold, respectively.C14-4 is one of the ionizable lipids to efficiently deliver mRNA to Jurkat cells or primary human T cells. It will effectively promote the development of mRNA delivery by iLNPs for CAR-T therapy.
DC33636	DOTAP	DOTAP, also known as 1,2-Dioleoyl-3-trimethylammoniumpropane, is a cationic liposome-forming compound used for transfection of DNA, RNA, and other negatively charged molecules into eukaryotic cells. It has been used in gene delivery vectors for gene ther
DC58047	DSPE-PEG 2000	PEG2000-DSPE is used for creating micelles that are able to carry drugs with low solubility.
DC31000	LP-01	LP-01 is an ionizable cationic amino lipid (pKa = ~6.1). It has been used in the generation of lipid nanoparticles (LNPs). LNPs containing LP-01 and encapsulating both Cas9 mRNA and modified single-guide RNA (sgRNA) for the transport protein transthyretin (Ttr) induce gene editing in liver cells in mice in a dose-dependent manner resulting in reduced serum Ttr levels for at least 12 months.