| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 2mg |
|
||
| 5mg |
|
||
| 10mg |
|
||
| Other Sizes |
| Targets |
Ki: 4.9 pM (hCD73)[1] IC50: 43 pM (Soluble hCD73)[1]
The primary target of Quemliclustat is CD73, a cell surface enzyme that converts extracellular AMP to adenosine. Adenosine is an immunosuppressive molecule that inhibits T cell and NK cell function in the tumor microenvironment. By inhibiting CD73, Quemliclustat reduces adenosine production, relieving immunosuppression and enhancing antitumor immunity. |
|---|---|
| ln Vitro |
AB-680 (AB680) suppresses soluble hCD73 at an IC50 of 0.043 nM, hCD73 in CHO cells, and CD73 in human and murine CD8+ T cells and hPBMC at IC50s of 0.070, 0.008, 0.66, and 0.011 nM, respectively [1].
In vitro, Quemliclustat is a potent and selective inhibitor of CD73, with an IC50 of 0.043 nM and a Ki of 4.9 pM. It is selective over other nucleotidases like NTPDase 2, 3, and 8 (IC50s >10 µM). Its activity is assessed by measuring the inhibition of AMP hydrolysis to adenosine in biochemical and cell-based assays. |
| ln Vivo |
On the basis of the extraordinary potency of AB680 in vitro and excellent pharmacokinetic profile in preclinical species, AB680 was advanced to a phase I clinical study in healthy volunteers. This placebo-controlled study assessed the safety, tolerability, and PK/PD profile of AB680. Here we present the initial pharmacokinetic data of AB680 in humans. AB680 was well tolerated when administered as a single iv dose across the range of 0.1–25 mg (six cohorts). All treatment-emergent adverse events were mild or moderate in severity, with no clear pattern of toxicity at these dose levels. AB680 displayed low clearance and a long half-life (67–74 h) following an iv infusion of 16 or 25 mg over 30–60 min. The human PK profile of AB680 is consistent with the intended Q2W dosing schedule and validates the design strategy employed to discover a long-acting drug for iv-administration.[1]
In vivo, Quemliclustat has anti-tumor activity. It is being investigated for the treatment of pancreatic ductal adenocarcinoma, metastatic prostate cancer, and other cancers. By inhibiting CD73, it enhances the immune response against tumors. Further studies are ongoing to evaluate its clinical efficacy. |
| Enzyme Assay |
CYP inhibition procedure: [1]
Test compounds were evaluated in-vitro for its potential to inhibit major human drug metabolizing enzymes of the cytochrome P450 family. The test compounds were incubated separately over a concentration range of 0 to 40 µM with 0.1 mg/mL human liver microsomal protein suspension in 0.1 M potassium phosphate buffer at pH 7.4, 1 mM NADPH, and a probe substrate (Phenacetin for CYP1A2, Diclofenac for CYP2C9, S-Mephenytoin for CYP2C19, Dextromethorphan for CYP2D6, and Midazolam for CYP3A4). Each substrate was incubated at 37 °C for 5 to 20 minutes as defined by the previous assay validation. Samples for each substrate were collected and pooled with samples from other substrate incubations for determination of product formation by LC-MS/MS. IC50 values were calculated using variable slope (4-parameter) model. Furafylline (1A2), sulfaphenazole (2C9), (+)-N-3-Benzylnirvanol (2C19), quinidine (2D6), and ketoconazole (3A4) were used as reference controls. For CD73 inhibition assays, the enzymatic activity of recombinant CD73 is measured. The enzyme is incubated with varying concentrations of Quemliclustat and a substrate (AMP). The production of adenosine or inorganic phosphate is measured. IC50 and Ki values are calculated. For selectivity, the compound is tested against other nucleotidases. |
| Cell Assay |
Cellular Human CD73 Assay. [1]
Generation and Expansion of Human CD73 Stable CHO Cell Line. Stable cell lines were generated by Lake Pharma using a standard protocol to transform CHO cells with a pcDNA3.1(+) vector carrying the human NT5E (CD73) gene. Antibiotic selection was performed in CD OptiCHO cell media containing 5 μg / mL Puromycin and 200 μg / mL Hygromycin B. Pools of surviving CHO-CD73 were collected and frozen in 7.5% DMSO in CD OptiCHO cell media. Cryopreserved cells were defrosted in a water bath at 37°C by agitating the vial until the cells were completely thawed. Cells were then transferred to a 15 mL Falcon tube prior to centrifuging at 225xg for 5 minutes to pellet the cells. The cell pellet was resuspended in fresh warm CD OptiCHO Growth Medium supplemented with 2 mM Glutamax and transferred to a T175 flask. After two days and on reaching ~80% confluence (~20 million cells/flask) cells were split 1:3 into three fresh T175 flasks. After a further three days, cells were transferred to a 15 mL Falcon tubes and centrifuged at 250xg for 5 minutes to pellet. Cells were re-suspended at a density of 3 million cells per mL in CellBanker2 cryopreservation media and aliquoted into cryogenic vials. Cell aliquots were stored at -80°C until needed. For cellular studies, cancer cells or immune cells are cultured. Quemliclustat is dissolved in DMSO and diluted in culture medium. Cells are treated with the compound. Adenosine levels in the supernatant are measured by LC-MS or an ELISA-based assay. T cell proliferation and cytotoxicity are assessed in co-culture assays. |
| Animal Protocol |
Clinical Study[1]
The phase I clinical study was a first-in-human, double-blind, randomized, placebo-controlled combined single-ascending-dose (SAD) and multiple-ascending-dose (MAD) study to evaluate the safety, tolerability, PK, and potential PD effects of AB680 in healthy volunteers. Participants were randomly selected to receive AB680 (n = 6) or matching placebo (n = 2) in each of seven dosing cohorts in the SAD part and a single dose cohort in the MAD part. In the SAD part of the study, participants received a single iv infusion of 0.1, 0.6, 2, 4, 8, 16, 25 mg of AB680 or placebo. In the MAD part of the study, participants received iv infusion of 8 mg of AB680 or placebo once daily on 3 days (days 1, 8, and 15). For in vivo efficacy studies, immunocompetent mice are implanted with syngeneic tumors. Quemliclustat is formulated in vehicle and administered intraperitoneally or orally. Tumor growth is monitored. Immune cell infiltration and activation in the tumor microenvironment are analyzed by flow cytometry. Adenosine levels in the tumor are measured. |
| ADME/Pharmacokinetics |
The pharmacokinetics (PK) of AB680 were evaluated in rodents and non-rodents. Preclinical animal models showed that AB680 exhibited extremely low clearance and a long half-life, suggesting that its PK characteristics in humans were suitable for intravenous (iv) administration, with a dosing regimen consistent with that of typical monoclonal antibody (mAb) dosing cycles. Preclinical animal models tolerated high-dose AB680 infusions well. [2]
Pharmacokinetic data for Quemliclustat indicate that it is a small-molecule inhibitor with favorable PK properties. Its half-life, Cmax, and AUC have been characterized in preclinical studies. Its PK profile supports its use in in vivo efficacy studies and clinical development. |
| Toxicity/Toxicokinetics |
Toxicological data for Quemliclustat are being evaluated in clinical studies. As an immunomodulatory agent, it may have immune-related adverse effects. Its safety profile is being characterized in ongoing trials. As with all drugs, appropriate safety precautions should be taken during handling.
|
| References | |
| Additional Infomation |
Quemliclustat is a small-molecule competitive inhibitor that inhibits the extracellular enzyme CD73 (differentiation cluster 73; 5'-extracellular nucleotidase; 5'-NT; extracellular 5'-nucleotidase), possessing potential immunomodulatory and antitumor activity. Upon administration, quemliclustat targets and binds to CD73, leading to CD73 aggregation and internalization. This prevents CD73-mediated conversion of adenosine monophosphate (AMP) to adenosine and reduces the level of free adenosine in the tumor microenvironment (TME). This prevents adenosine-mediated lymphocyte suppression and enhances the activity of CD8-positive effector cells and natural killer (NK) cells. Furthermore, it activates macrophages and reduces the activity of myeloid-derived suppressor cells (MDSCs) and regulatory T lymphocytes (Tregs). CD73 inhibits tumor cell growth by eliminating the suppression of the immune system and enhancing cytotoxic T cell-mediated anti-cancer immune responses. In addition, CD73 aggregation and internalization reduce the migration ability of cancer cells and prevent metastasis. CD73 is a plasma membrane protein belonging to the 5'-nucleotidase (NTase) family. It is upregulated in various cancer cell types and catalyzes the conversion of extracellular nucleotides (such as AMP) into membrane-permeable nucleosides (such as adenosine). In the tumor microenvironment (TME), CD73 plays a key role in adenosine-mediated immunosuppression.
Quemliclustat (CAS 2105904-82-1) is a potent CD73 inhibitor with an IC50 of 0.043 nM and a Ki of 4.9 pM. It has the molecular formula C₂₀H₂₄ClFN₄O₉P₂. It has anti-tumor activity and is being investigated for cancer treatment. It is strictly for research use. |
| Molecular Formula |
C20H24CLFN4O9P2
|
|---|---|
| Molecular Weight |
580.824889183044
|
| Exact Mass |
580.069
|
| Elemental Analysis |
C, 41.36; H, 4.17; Cl, 6.10; F, 3.27; N, 9.65; O, 24.79; P, 10.67
|
| CAS # |
2105904-82-1
|
| Related CAS # |
AB-680 ammonium
|
| PubChem CID |
130205852
|
| Appearance |
White to off-white solid powder
|
| LogP |
-0.7
|
| Hydrogen Bond Donor Count |
6
|
| Hydrogen Bond Acceptor Count |
13
|
| Rotatable Bond Count |
9
|
| Heavy Atom Count |
37
|
| Complexity |
893
|
| Defined Atom Stereocenter Count |
5
|
| SMILES |
ClC1=CC(=C2C=NN(C2=N1)[C@H]1[C@@H]([C@@H]([C@@H](COP(CP(=O)(O)O)(=O)O)O1)O)O)N[C@@H](C)C1C=CC=CC=1F
|
| InChi Key |
MFYLCAMJNGIULC-KCVUFLITSA-N
|
| InChi Code |
InChI=1S/C20H24ClFN4O9P2/c1-10(11-4-2-3-5-13(11)22)24-14-6-16(21)25-19-12(14)7-23-26(19)20-18(28)17(27)15(35-20)8-34-37(32,33)9-36(29,30)31/h2-7,10,15,17-18,20,27-28H,8-9H2,1H3,(H,24,25)(H,32,33)(H2,29,30,31)/t10-,15+,17+,18+,20+/m0/s1
|
| Chemical Name |
(((((2R,3S,4R,5R)-5-(6-chloro-4-(((S)-1-(2-fluorophenyl)ethyl)amino)-1H-pyrazolo[3,4-b]pyridin-1-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic
acid
|
| Synonyms |
AB680; Quemliclustat; AB 680; Quémliclustat; AB-680; Quemliclustat; 2105904-82-1; AB680; Quemliclustat [USAN]; UNII-J6K8WSV73A; J6K8WSV73A; QUEMLICLUSTAT [INN]; Quemliclustatum; AB-680
|
| 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 Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture. |
| 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 : ~200 mg/mL (~344.34 mM)
H2O : ~5 mg/mL (~8.61 mM) |
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 7.5 mg/mL (12.91 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 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 75.0 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. Solubility in Formulation 2: ≥ 7.5 mg/mL (12.91 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 75.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 7.5 mg/mL (12.91 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. Solubility in Formulation 4: ≥ 2.5 mg/mL (4.30 mM) in 5% DMSO + 40% PEG300 + 5% Tween80 + 50% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 5: 2.5 mg/mL (4.30 mM) (saturation unknown) in 5% DMSO + 95% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Solubility in Formulation 6: ≥ 0.5 mg/mL (0.86 mM) (saturation unknown) in 1% DMSO 99% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.7217 mL | 8.6085 mL | 17.2170 mL | |
| 5 mM | 0.3443 mL | 1.7217 mL | 3.4434 mL | |
| 10 mM | 0.1722 mL | 0.8609 mL | 1.7217 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
