| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
QPCTL (glutaminyl-peptide cyclotransferase-like protein) – IC50 = 3.8 ± 0.7 nM. [1]
QPCT (glutaminyl-peptide cyclotransferase) – comparable inhibition to QPCTL (exact IC50 not provided in main text; supplementary Fig. S3 shows dose-response curve). [1] |
|---|---|
| ln Vitro |
QP5038 inhibited CD47 pyroglutamation (pGlu-CD47) in HEK293T cells with an IC50 of 3.3 ± 0.5 nM, as determined by dose-dependent inhibition assay. [1]
QP5038 attenuated the binding of CD47 to SIRPα in a dose-dependent manner in HEK293T cells with an IC50 of 8.5 ± 4.9 nM, more potent than PQ912 and SEN177. [1] In HEK293T and Raji cells, QP5038 (100 nM) showed superior inhibitory effects on CD47 pyroglutamation compared to SEN177 and PQ912, without altering overall cell surface levels of CD47. [1] QP5038 significantly attenuated CD47 pyroglutamation in various cancer cell lines including B16F10, HCT116, SKOV3, Huh7, T24, MCF-6, SU-DHL-8, H929, and H1299. [1] In an in vitro phagocytosis assay, QP5038 (10 μM, 48h treatment) significantly boosted macrophage-mediated phagocytosis of B16F10 cells in combination with anti-TRP1 antibody TA99, and of Raji cells in combination with rituximab. The phagocytosis efficiency of QP5038 was much better than that of SEN177 and PQ912. [1] QP5038 did not markedly impair cell viability in Raji or B16F10 cells (supplementary Fig. S5). [1] QP5038 did not attenuate the cell viability of bone marrow-derived macrophages (BMDM) nor their phagocytotic ability toward apoptotic Jurkat cells (supplementary Fig. S8). [1] In a senescence model, pretreatment with QP5038 blocked the inhibitory effect of senescent cells on macrophage-mediated efferocytosis of apoptotic cells (supplementary Fig. S7). [1] |
| ln Vivo |
In a B16F10 syngeneic mouse tumor model (C57BL/6 mice, subcutaneous injection of 1×10⁶ B16F10 cells), combination treatment of QP5038 (25 mg/kg, intraperitoneal, once daily) with anti-PD-1 antibody (80 μg/mouse, intraperitoneal, once weekly starting day 12) dramatically suppressed both tumor growth and tumor weight compared to each single treatment and to SEN177 treatment group, without effect on mouse body weight. [1]
In another B16F10 model (5×10⁶ cells), QP5038 (25 mg/kg, i.p., once daily from day 6) plus anti-PD-1 antibody showed superior anti-tumor efficacy compared to SEN177 plus anti-PD-1. [1] Depletion of T cells in mice using anti-CD3 antibody blocked the enhancement of QP5038 on anti-PD-1 antibody-mediated tumor inhibition, suggesting that the anti-cancer effect of QP5038 is due to activation of immune response. [1] |
| Enzyme Assay |
QPCTL or QPCT protein (7 ng/μL, 12.5 μL) and various concentrations of test compound (2.5 μL) were added to a 384-well plate on ice. The reaction was shaken at 37°C, 100 rpm for 10 minutes. Then, 10 μL of fluorescent substrate H-Gln-AMC (500 μM) was added and incubated at 100 rpm for 20 minutes at 37°C. Finally, 25 μL of PGPEP I (pyroglutamyl aminopeptidase I, 5.74 ng/μL) was added and incubated at 37°C, 100 rpm for 30 minutes. Fluorescence was measured at excitation 380 nm/emission 460 nm. IC50 values were calculated using GraphPad Prism 8.0. For QP5038, the IC50 against QPCTL was 3.8 ± 0.7 nM. [1]
PGPEP I protein was purified from E. coli BL21(DE3) cells transformed with pET28a vector containing human PGPEP I. Cells were induced with 1 mM IPTG at 20°C for 48 hours, lysed by high-pressure homogenization, and purified by Ni-NTA affinity chromatography followed by size exclusion chromatography. [1] QPCTL protein was expressed in E. coli BL21(DE3) cells using pGEX-4T-2 vector containing human QPCTL cDNA. Cells were induced with 1 mM IPTG at 20°C for 8-10 hours, lysed by ultrasonication, and purified using GST-affinity beads. [1] |
| Cell Assay |
HEK293T, B16F10, HCT116, SKOV3, Huh7, T24, MCF-6, SU-DHL-8, H929, Raji, H1299, and Jurkat cells were cultured in DMEM or RPMI 1640 with 10% FBS, penicillin (100 U/mL), and streptomycin (100 μg/mL) at 37°C in 5% CO2. [1]
For CD47 pyroglutamation assay: Cells were treated with QP5038 at indicated concentrations (e.g., 100 nM, or dose titration for IC50 determination). pGlu-CD47 levels and total CD47 surface levels were measured by flow cytometry using specific antibodies. [1] For SIRPα binding assay: Cells were incubated with PE-labeled recombinant human or mouse SIRPα protein for 1 hour on ice, protected from light, then analyzed by flow cytometry. QP5038 reduced SIRPα binding in a dose-dependent manner with IC50 of 8.5 ± 4.9 nM in HEK293T cells. [1] For phagocytosis assay: Cancer cells were stained with 5 μM CFSE at 37°C for 10 minutes. Macrophages (BMDM) were plated at 1×10⁵ cells per well in 48-well plates. Target cells and macrophages were co-cultured for 4 hours at 37°C. Macrophages were identified with APC-labeled anti-F4/80, and flow cytometry was performed. Phagocytosis (%) = [CFSE⁺F4/80⁺ cells / total CFSE⁺ cells] × 100%. QP5038 (10 μM, 48h pre-treatment of cancer cells) significantly boosted phagocytosis with TA99 or rituximab. [1] For efferocytosis assay: Senescent B16F10 cells were induced by 10 μM cisplatin for 4-7 days and confirmed by β-galactosidase staining. Apoptotic Jurkat cells were induced by 5 μM camptothecin for 4 hours. BMDMs were co-cultured with senescent cells for 16 hours, then CFSE-labeled apoptotic Jurkat cells were added at 1:2 ratio (macrophages:Jurkat) for 60 minutes. Efferocytosis was calculated as % of CFSE⁺F4/80⁺ cells among F4/80⁺ cells. QP5038 pretreatment blocked the inhibitory effect of senescent cells on efferocytosis. [1] For cell viability: Raji cells (5000/well), B16F10 cells (3000/well), or BMDM (10000/well) were plated in 96-well plates. Cell viability was examined using CCK-8 assay according to manufacturer's instructions. QP5038 did not markedly impair cell viability. [1] |
| Animal Protocol |
For B16F10 syngeneic tumor model (first): Male C57BL/6 mice were subcutaneously implanted with 1×10⁶ B16F10 cells in 100 μL DMEM on Day 0. Anti-PD-1 antibody (clone RMP1-14) or mouse IgG2a isotype control was dosed at 80 μg per mouse intraperitoneally starting on day 12 after tumor implantation, followed by once-weekly regimen. QP5038 was intraperitoneally administered at 25 mg/kg once daily. Tumors were measured daily, and mice were sacrificed 24 days after tumor challenge. Tumor volume = (length × width²) × 0.5. [1]
For comparison with SEN177: Male C57BL/6 mice were subcutaneously implanted with 5×10⁶ B16F10 cells in 100 μL DMEM on Day 0. Anti-PD-1 antibody or isotype control was dosed at 80 μg per mouse i.p. starting on day 6, once weekly. QP5038 or SEN177 was intraperitoneally administered at 25 mg/kg once daily from day 6. Tumors were measured every three days, and mice were sacrificed on day 15. Mouse organs and blood were collected for blood routine, serum biochemistry, and histological evaluation. [1] For T cell depletion study: 6-8 week old male C57BL/6 mice were subcutaneously injected with B16F10 cells (8.5×10⁵ cells/100 μL) into right flank. Anti-CD3 antibody (200 μg/mouse) or isotype control was injected every 4 days starting one day before tumor implantation (Day -1). When tumor volume reached ~50 mm³, mice received intraperitoneal injections of anti-PD-1 antibody (80 μg/mouse once weekly) and QP5038 (25 mg/kg once daily) or vehicle control. Animal weights and tumor volumes measured twice weekly. T cell depletion confirmed by flow cytometry with PE-anti-mouse CD3 antibody. [1] |
| Toxicity/Toxicokinetics |
In vivo toxicity assessment in mice showed that QP5038 (25 mg/kg, daily for 15 days) did not cause abnormal blood composition (WBC, RBC, PLT, LYM, MON, NEU) or organ damage (liver, kidney, spleen, heart) as assessed by hematoxylin and eosin (HE) staining. Blood serum biochemistry (creatinine, ALT, AST, BUN) showed no significant differences compared to control vehicle. [1]
QP5038 did not markedly impair cell viability of Raji cells, B16F10 cells, or bone marrow-derived macrophages (BMDM) in vitro. [1] QP5038 did not attenuate the phagocytotic ability of macrophages toward apoptotic cells. [1] |
| References | |
| Additional Infomation |
QP5038 is a benzonitrile-based QPCTL inhibitor developed via structure-based optimization from SEN177. The nitrile group in the benzonitrile core forms a hydrogen bond with Glu325 residue in the binding model. QP5038 inhibits CD47 pyroglutamation, blocks CD47/SIRPα interaction, and promotes macrophage-mediated phagocytosis of cancer cells. [1]
The anti-cancer effect of QP5038 in combination with anti-PD-1 antibody is dependent on T cells, as T cell depletion using anti-CD3 antibody blocked the enhanced tumor inhibition. [1] QP5038 also blocked the inhibitory effect of senescent cells on macrophage-mediated efferocytosis of apoptotic cells. [1] |
| Molecular Formula |
C21H20FN5
|
|---|---|
| Molecular Weight |
361.42
|
| Exact Mass |
361.170273
|
| CAS # |
3031429-88-3
|
| PubChem CID |
169494342
|
| Appearance |
Off-white to yellow solid powder
|
| LogP |
3.1
|
| Hydrogen Bond Donor Count |
0
|
| Hydrogen Bond Acceptor Count |
5
|
| Rotatable Bond Count |
3
|
| Heavy Atom Count |
27
|
| Complexity |
540
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
CN1C=NC=C1C2CCN(CC2)C3=C(C=CC=C3C4=CN=C(C=C4)F)C#N
|
| InChi Key |
IUBVPNGSCDYBJU-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C21H20FN5/c1-26-14-24-13-19(26)15-7-9-27(10-8-15)21-16(11-23)3-2-4-18(21)17-5-6-20(22)25-12-17/h2-6,12-15H,7-10H2,1H3
|
| Chemical Name |
3-(6-fluoro-3-pyridinyl)-2-[4-(3-methylimidazol-4-yl)piperidin-1-yl]benzonitrile
|
| Synonyms |
QP5038; QP 5038; QP-5038; 3031429-88-3; orb3133289; SCHEMBL29195008; GTPL13401;
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| Solubility (In Vitro) |
DMSO：～100 mg/mL (276.7 mM)
|
|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.7669 mL | 13.8343 mL | 27.6686 mL | |
| 5 mM | 0.5534 mL | 2.7669 mL | 5.5337 mL | |
| 10 mM | 0.2767 mL | 1.3834 mL | 2.7669 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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