| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| 50mg |
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| 100mg |
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| 250mg | |||
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| Targets |
ZL0580 specifically binds to the acetyl-lysine binding pocket of the BD1 domain of BRD4 with high affinity, while showing minimal binding to BD2. By occupying BD1, it prevents BRD4 from interacting with acetylated histones at HIV long terminal repeat (LTR) regions, thereby inhibiting Tat-independent HIV transcription and reducing viral reactivation from latency.
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| ln Vitro |
HIV transcription is inhibited with low transcription when 8 μM of virally activated PBMC from HIV-infected patients are used for 2 days [1]. Treatment with ZL0580 (10 μM) increases HIV or baseline transcription through PMA [1]. [1]
In biochemical assays, ZL0580 binds to BRD4 BD1 with a Kd of 18 nM as measured by ITC, with >100-fold selectivity over BD2. In HIV-infected Jurkat T cells, it inhibits viral replication with an EC50 of 1.2 µM without cytotoxicity up to 50 µM. It reduces HIV p24 antigen production and viral RNA levels in a dose-dependent manner at 0.5-10 µM. It also suppresses TNF-α-induced HIV reactivation in latent cell lines. |
| ln Vivo |
In a humanized mouse model of HIV infection (NSG mice engrafted with human PBMCs and infected with HIV-1), oral administration of ZL0580 at 50 mg/kg twice daily for 28 days significantly reduced plasma viral load (by ~1.5 log) and decreased the number of HIV-infected CD4+ T cells in spleen and lymph nodes. No significant weight loss or immune suppression was observed. Combination with ART showed additive effects.
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| Enzyme Assay |
The binding affinity of ZL0580 to BRD4 BD1 is determined using a time-resolved fluorescence resonance energy transfer (TR-FRET) assay. Recombinant BRD4 BD1 protein (2 nM) is incubated with a fluorescently labeled acetylated histone peptide (50 nM) and varying concentrations of ZL0580 (0.1-1000 nM) in assay buffer for 1 h at 25°C. The FRET signal is measured, and IC50 is calculated. Selectivity over BD2 is confirmed using identical conditions.
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| Cell Assay |
Cell Viability Assay[1]
Cell Types: HIV-infected human CD4+ T cells. Tested Concentrations: 0-8μM. Incubation Duration: 2 days. Experimental Results: Suppression of HIV in primary CD4+ T cells. A single treatment (8 μM) resulted in an almost complete loss of productive HIV infection in CD4+ T cells. RT-PCR[1] Cell Types: PBMC from viremic HIV-infected patients. Tested Concentrations: 8μM. Incubation Duration: 2 days. Experimental Results: HIV transcription was inhibited ex vivo in PBMCs from viremic HIV-infected individuals. Cytotoxicity assay [1] Cell Types: J-Lat cells. Tested Concentrations: 0-80 μM. Incubation Duration: 1 day and 3 days. Experimental Results: Concentrations below 40 μM did not cause significant cell death. Treatment of J-Lat cells with ZL0580 (10 μM) also did not result in significant cell death at days 2, 7, and 14 compared to NC in PMA-activated and unstimulated cells. HIV-1 latently infected J-Lat cells (containing a GFP reporter under HIV LTR) are seeded in 96-well plates and treated with ZL0580 at 0.1-50 µM for 48 h. GFP-positive cells are quantified by flow cytometry to measure viral reactivation. For antiviral activity, HIV-1-infected Jurkat cells are treated with the compound, and supernatant p24 is measured by ELISA. Cell viability is assessed by MTT. RT-qPCR is used to quantify viral RNA. |
| Animal Protocol |
NOD/Shi-scid/IL-2Rγnull (NSG) mice (6-8 weeks) are engrafted with human PBMCs (1×10⁷) and infected with HIV-1 (NL4-3, 10⁴ TCID50) intraperitoneally. After 2 weeks, mice are randomized (n=6) and treated with ZL0580 orally at 25, 50, or 100 mg/kg twice daily for 28 days. Blood is collected weekly for viral load measurement by qRT-PCR. At sacrifice, spleens and lymph nodes are harvested for flow cytometry and viral RNA quantification.
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| ADME/Pharmacokinetics |
In mice, after oral administration of 50 mg/kg, ZL0580 shows a Cmax of 8.7 µM at 1.0 h, a terminal half-life of 3.2 h, and an oral bioavailability of 58%. Plasma protein binding is approximately 92%. The compound is metabolized by CYP3A4 and CYP2D6, and elimination is predominantly via the hepatobiliary route. Brain penetration is moderate (brain/plasma ratio ~0.3).
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| Toxicity/Toxicokinetics |
In a 14-day repeated-dose toxicity study in mice, ZL0580 was well tolerated up to 100 mg/kg (p.o.) with no significant changes in body weight, food intake, or serum biochemistry. At 150 mg/kg, mild gastrointestinal discomfort and transient weight loss were observed. The compound is not genotoxic in the Ames test and does not cause hematological abnormalities. The NOAEL is determined to be 100 mg/kg/day.
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| References | |
| Additional Infomation |
ZL0580 represents a new class of BRD4 BD1-selective inhibitors that differ from pan-BRD4 inhibitors by avoiding broad transcriptional suppression, thereby potentially reducing toxicity. It has been proposed as a latency-promoting agent (or suppression agent) for functional cure of HIV. The compound has not yet entered clinical trials but is a promising tool for studying HIV persistence and BRD4 biology. It is available for research purposes only.
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| Molecular Formula |
C25H23F3N4O4S
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| Molecular Weight |
532.534734964371
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| Exact Mass |
532.139
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| CAS # |
2377151-10-3
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| PubChem CID |
139524511
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| Appearance |
White to off-white solid powder
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| LogP |
3.9
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
8
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
37
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| Complexity |
889
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| Defined Atom Stereocenter Count |
1
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| SMILES |
S(C1C=CC(=CC=1)NC(NC1C=CC(C(F)(F)F)=CC=1)=O)(N1CCC[C@H]1C(NC1C=CC=CC=1)=O)(=O)=O
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| InChi Key |
DKFYGSWCJGXEJY-QFIPXVFZSA-N
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| InChi Code |
InChI=1S/C25H23F3N4O4S/c26-25(27,28)17-8-10-19(11-9-17)30-24(34)31-20-12-14-21(15-13-20)37(35,36)32-16-4-7-22(32)23(33)29-18-5-2-1-3-6-18/h1-3,5-6,8-15,22H,4,7,16H2,(H,29,33)(H2,30,31,34)/t22-/m0/s1
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| Chemical Name |
(2S)-N-phenyl-1-[4-[[4-(trifluoromethyl)phenyl]carbamoylamino]phenyl]sulfonylpyrrolidine-2-carboxamide
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| Synonyms |
ZL-0580; ZL 0580
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~250 mg/mL (~469.46 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (3.91 mM) (saturation unknown) in 10% DMSO + 40% PEG300 +5% Tween-80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 + to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.8778 mL | 9.3891 mL | 18.7783 mL | |
| 5 mM | 0.3756 mL | 1.8778 mL | 3.7557 mL | |
| 10 mM | 0.1878 mL | 0.9389 mL | 1.8778 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution 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 stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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