H-Leu-Leu-OMe . HBr (Synonyms: LLOMe hydrobromide; Leu-Leu methyl ester hydrobromide; H-Leu-Leu-OMe hydrobromide) |
Catalog No.GA22634
One-Click Copy Product Info
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H-Leu-Leu-OMe . HBr (Leu-Leu methyl ester hydrobromide) is a dipeptide generated by human monocytes or polymorphonuclear leukocytes. H-Leu-Leu-OMe . HBr selectively eliminates lymphocytes, especially natural killer cells with cytotoxic potential, and induces endolysosomal pathway stress
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 16689-14-8
Sample solution is provided at 25 µL, 10mM.
H-Leu-Leu-OMe . HBr (Leu-Leu methyl ester hydrobromide) is a dipeptide generated by human monocytes or polymorphonuclear leukocytes. H-Leu-Leu-OMe . HBr selectively eliminates lymphocytes, especially natural killer cells with cytotoxic potential, and induces endolysosomal pathway stress[1]. The sensitivity of cytotoxic lymphocytes to H-Leu-Leu-OMe . HBr depends upon the production of membranolytic metabolites of the structure (Leu-Leu)n-OMe by dipeptidyl peptidase I[2]. Such membranolytic metabolites are present at far higher levels in cytotoxic lymphocytes than in cells without cytolytic potential or not of bone marrow origin[2].
In vitro, H-Leu-Leu-OMe . HBr may act as an inflammasome activator on RPE cells (primary human fetal RPE cells; 1mM; 3h), leading to caspase-1 cleavage, cytokine secretion, and X-linked inhibitor of apoptosis protein (XIAP) reduction[3]. H-Leu-Leu-OMe . HBr activates NLRP3 inflammasome that promotes IL-6 and IL-8 production (human umbilical vein endothelial cells, 20μM; 19h) in primed human umbilical vein endothelial cells[4]. Moreover, H-Leu-Leu-OMe . HBr may act as an apoptosis inducer (U937 cells; 0.5mM; 30min) in natural killer cells and macrophages[5].
In vivo, H-Leu-Leu-OMe . HBr shows toxicity to lymphocytes (adult (CBA x BALB/c) F1 mice, H-Leu-Leu-OMe . HBr-treated CBA spleen cells(0.15M; 15min; room temperature; i.p) that suppressing lymphocyte function[6]. H-Leu-Leu-OMe . HBr can also mimic alum-mediated necrosis that specifically kills myeloid leukocytes in 8-week-old female C57BL/6 mice (1mg; once; i.v)[7].
References:
[1] Thiele, D L, and P E Lipsky. “The immunosuppressive activity of L-leucyl-L-leucine methyl ester: selective ablation of cytotoxic lymphocytes and monocytes.” *Journal of immunology (Baltimore, Md. : 1950)* vol. 136,3 (1986): 1038-48.
[2] Thiele, D L, and P E Lipsky. “Mechanism of L-leucyl-L-leucine methyl ester-mediated killing of cytotoxic lymphocytes: dependence on a lysosomal thiol protease, dipeptidyl peptidase I, that is enriched in these cells.” *Proceedings of the National Academy of Sciences of the United States of America* vol. 87,1 (1990): 83-7. doi:10.1073/pnas.87.1.83
[3] Gao, Jiangyuan et al. “The reduction of XIAP is associated with inflammasome activation in RPE: implications for AMD pathogenesis.” Journal of neuroinflammation vol. 16,1 171. 22 Aug. 2019, doi:10.1186/s12974-019-1558-5
[4] Kinnunen, K et al. “Lysosomal destabilization activates the NLRP3 inflammasome in human umbilical vein endothelial cells (HUVECs).” *Journal of cell communication and signaling* vol. 11,3 (2017): 275-279. doi:10.1007/s12079-017-0396-4
[5] Odaka, M et al. “Synthesis, photoreactivity and cytotoxic activity of caged compounds of L-leucyl-L-leucine methyl ester, an apoptosis inducer.” *Photochemistry and photobiology* vol. 63,6 (1996): 800-6. doi:10.1111/j.1751-1097.1996.tb09633.x
[6] Mowat, A M, and P A Leck. “Generalized toxicity of L-leucyl-leucine-methyl ester for lymphocyte functions.” *Immunology* vol. 69,4 (1990): 564-9.
[7]Jacobson, Lee S et al. “Cathepsin-mediated necrosis controls the adaptive immune response by Th2 (T helper type 2)-associated adjuvants.” *The Journal of biological chemistry* vol. 288,11 (2013): 7481-7491. doi:10.1074/jbc.M112.400655
| Cell experiment [1]: | |
Cell lines | primary human fetal RPE cells |
Preparation Method | L-leucyl-L-leucine methyl ester (H-Leu-Leu-OMe . HBr) was applied with or without the pre-incubation of recombinant human IL-1α at 10ng/mL (“priming”). Leu-Leu-OMe concentration of 1mM was proven effective to destabilize RPE lysosomes according to previously established protocol. After 3h of H-Leu-Leu-OMe . HBr stimulation (1mL/well at 1mM), cell culture supernatants were collected and cells were lysed in 200μL/well ice-cold RIPA buffer supplemented with protease inhibitors. |
Reaction Conditions | 1mM, 3h |
Applications | In RPE cell culture, H-Leu-Leu-OMe . HBr stimulation led to caspase-1 cleavage, cytokine secretion, and XIAP reduction, which can be abolished by Z-YVAD-FMK. |
| Animal experiment [2]: | |
Animal models | adult (CBA x BALB/c) F1 mice |
Preparation Method | Cells to be treated were incubated with H-Leu-Leu-OMe . HBr at 2.5*106cell/ml for 15min at room temperature. Adult (CBA x BALB/c) F1 mice were injected intraperitoneally with 6*107 CBA spleen cells. |
Dosage form | H-Leu-Leu-OMe . HBr-treated CBA spleen cells(0.15M, 15min, room temperature), i.p |
Applications | H-Leu-Leu-OMe . HBr prevented immune lymphocytes from mediating both class I and class II major histocompatibility complex (MHC)-restricted delayed-type hypersensitivity (DTH) responses in vivo. |
References: | |
| Cas No. | 16689-14-8 | SDF | |
| Synonyms | LLOMe hydrobromide; Leu-Leu methyl ester hydrobromide; H-Leu-Leu-OMe hydrobromide | ||
| Formula | C13H26N2O3 · HBr | M.Wt | 339.27 |
| Solubility | DMSO : 125 mg/mL (368.44 mM; Need ultrasonic); H2O : 100 mg/mL (294.75 mM; Need ultrasonic) | Storage | Store at -20°C |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 2.9475 mL | 14.7375 mL | 29.475 mL |
| 5 mM | 589.5 μL | 2.9475 mL | 5.895 mL |
| 10 mM | 294.8 μL | 1.4738 mL | 2.9475 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
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Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL saline, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >97.00% Appearance: A solid
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 12 reference(s) in Google Scholar.)