GLUT1 (IC50 = 115 nM); GLUT2 (IC50 = 137 nM); GLUT3 (IC50 = 90 nM); GLUT4 (IC50 = 68 nM)

KL-11743 (compound 8) has IC50s of 33 nM and 268 nM at 0.37 mM and 10 mM glucose, respectively, and competes with glucose for binding to GLUT1 [1]. HT-1080 cell proliferation is dose-dependently inhibited by KL-11743 (39-10000 nM; 24-72 h), with an IC50 of 677 nM [3]. KL-11743 has a more potent inhibitory effect on the proliferation of KEAP1 mutant lung cancer cells in comparison to KEAP1-WT lung cancer cells [4]. In HT-1080 cells, KL-11743 (0.001-10 μM) causes a rapid rise in AMPK and acetyl-CoA carboxylase phosphorylation [3]. In 786-O cells, KL-11743 (2 μM) decreases the absorption of glucose. NADP+/NADPH is increased in NCl-H226 cells by KL-11743. In SLC7A11 high cancer cell lines (NCl-H226 and UMRC6 cells), KL-11743 causes cell death [2]. KL-11743 (0.001-10 μM) totally suppresses glycolytic ATP synthesis in HT-1080 fibrosarcoma cells and inhibits glucose consumption, lactate secretion, and 2DG transport with IC50 values of 228, 234, and 87 nM, respectively. 127 nM is the cellular IC50 [3].

KL-11743 (100 mg/kg; intraperitoneally every two days for 5 weeks) reduces the growth of SLC7A11 high NCI-H226 xenograft tumors and is well tolerated in vivo [2]. KL-11743 (30-100 mg/kg; single oral dose) significantly increases blood glucose levels and delays glucose clearance in mice challenged with 5 g/kg glucose [3]. KL-11743 significantly inhibits the growth of KEAP1 KO tumors [4]. Plasma levels of KL-11743 (100 mg/kg; i.p.) were maintained at inhibitory levels during most of the 24-hour dosing period [2]. KL-11743 (oral) exhibits moderate oral concentrations between 30% and 15% in mice (10-100 mg/kg) and rats (10-300 mg/kg), and good dose-linear plasma exposure curve, reaching a concentration of approximately 20 μM) [3]. KL-11743 exhibited comparable half-lives in rats, ranging from 2.04 to 5.38 hours (10 mg/kg intravenously; 10-300 mg/kg orally) and 1.45-4.75 hours in mice (iv and Intraperitoneal injection 10 mg/kg); 10-100 mg/kg (orally) [3].

GLUT, GLUT1, and GLUT3 Assays[1]
GLUT1 and GLUT3 assays were performed using a modified version of the pairwise assay described by Siebeneicher et al. Briefly, GLUT1-dependent DLD1 wild-type and GLUT3-dependent DLD1-SLC2A1–/– were maintained in RPMI 1640 cell culture medium containing 10% fetal bovine serum, 1% penicillin–streptomycin, and 10 mM HEPES in a humidified incubator with 5% CO2 at 37 °C. The day before the assay, the cells were seeded in 90 μL of this medium in 96-well plates at a density of 50 000 cells/well and allowed to attach overnight. The day of the assay, 10 μL of the culture medium containing GLUT inhibitors and oligomycin was added to each well to a final concentration of 10 μM oligomycin and 0.06% DMSO. The plates were returned to the incubator for 90 min, and then ATP levels were determined using the CellTiter-Glo Luminescent Cell Viability Assay. For the GLUT HT-1080 assay, the procedure was the same except the cells were seeded at 40 000 cells/well.

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Glucose Competition Assay[1]
The assay was performed according to the standard GLUT1 (DLD-1) assay protocol, except that the cells were washed once in 100 μL of PBS prior to the addition of media containing the test compounds, oligomycin (10 μM), and glucose at either 10, 3.33, 1.11, or 0.37 mM concentrations.

Cell viability assay [3]
Cell Types: HT-1080 Cell
Tested Concentrations: 39, 78, 156, 312, 625, 1250, 2500, 5000, 10000 nM
Incubation Duration: 24, 48, 72 hrs (hours)
Experimental Results: Inhibition of the growth of HT-1080 cells in a dose-dependent manner.

Animal/Disease Models: 4 to 6 weeks old athymic nude mice (Foxn1nu/Foxn1nu) were injected with NCI-H226 cells at 100 mg/kg[2]
Doses: 100 mg/kg
Route of Administration: intraperitoneal (ip) injection every two days for 5 weeks.
Experimental Results: Inhibit tumor growth. Exhibits extensive necrotic cell death. The level of the PPP intermediate 6-phosphogluconate decreases and the NADP+/NADPH ratio increases.

[1]. Discovery and Optimization of Glucose Uptake Inhibitors. J Med Chem. 2020 May 28;63(10):5201-5211.

[2]. Cystine transporter regulation of pentose phosphate pathway dependency and disulfide stress exposes a targetable metabolic vulnerability in cancer. Nat Cell Biol. 2020 Apr;22(4):476-486.

[3]. Inhibition of glucose transport synergizes with chemical or genetic disruption of mitochondrial metabolism and suppresses TCA cycle-deficient tumors. Cell Chem Biol. 2021 Oct 22;S2451-9456(21)00441-4.

[4]. KEAP1 deficiency drives glucose dependency and sensitizes lung cancer cells and tumors to GLUT inhibition. iScience. 2021 May 25;24(6):102649.

Aerobic glycolysis was first discovered by Warburg and is considered a hallmark of cancer. In recent years, it has also been considered to be associated with the activation and proliferation of immune cells. Glucose is the starting material for glycolysis, which crosses the cell membrane via a class of glucose transporters (GLUTs). Therefore, targeting glucose transporters to regulate aerobic glycolysis is an attractive strategy for finding potential therapeutics for cancer and autoimmune diseases. This article describes the discovery and optimization of a class of highly effective, orally bioavailable glucose transporter inhibitors that target both GLUT1 and GLUT3. [1]

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Research on targeting glucose metabolism in cancer has been limited due to the poor efficacy and specificity of existing anti-glycolysis drugs and the lack of understanding of glucose dependence in cancer subtypes in vivo. This article introduces a series of well-characterized, highly effective, orally bioavailable class I glucose transporter (GLUT) inhibitors. The representative compound KL-11743 specifically blocks glucose metabolism, leading to acute collapse of the NADH pool and significant accumulation of aspartate, indicating a significant shift in oxidative phosphorylation in mitochondria. Disrupting mitochondrial metabolism by chemically inhibiting electron transport, deleting endogenous mutations in the malate-aspartate shuttle component GOT1, or tricarboxylic acid cycle enzymes can lead to synthetic lethality with KL-11743. Patient-derived xenograft models of succinate dehydrogenase A (SDHA) deficient cancers are specifically sensitive to KL-11743, which directly demonstrates that TCA cycle mutant tumors are susceptible to GLUT inhibitors in vivo. [3] Metabolic reprogramming of cancer cells can lead to metabolic defects. KEAP1 mutant lung cancer is resistant to most existing therapies. This study shows that KEAP1 deficiency promotes glucose dependence in lung cancer cells, and KEAP1 mutant/deficient lung cancer cells are more susceptible to glucose deprivation than wild-type cells. Mechanistically, KEAP1 inactivation in lung cancer cells induces constitutive activation of NRF2 transcription factors and abnormal expression of the NRF2 target cystine transporter SLC7A11. Under glucose restriction, high cystine uptake in KEAP1-inactivated lung cancer cells stimulates the accumulation of toxic disulfide bonds, NADPH depletion, and cell death. Gene knockout of the NRF2-SLC7A11 axis or treatments that inhibit disulfide bond accumulation can salvage these consequences. Finally, we found that KEAP1-inactivated lung cancer cells or xenografts are sensitive to glucose transporter inhibitors. In summary, our findings reveal that KEAP1 deficiency induces glucose dependence in lung cancer cells and identify a therapeutically significant metabolic defect. [4]

C30H30N6O3

522.597606182098

522.24

C, 68.95; H, 5.79; N, 16.08; O, 9.18

1369452-53-8

146345838

Light yellow to green yellow solid powder

5.5

3

7

10

39

763

0

XKOYTLRGOQTKAU-UHFFFAOYSA-N

InChI=1S/C30H30N6O3/c1-4-38-25-12-13-27-26(15-25)30(34-23-10-8-20(9-11-23)22-16-31-32-17-22)36-29(35-27)21-6-5-7-24(14-21)39-18-28(37)33-19(2)3/h5-17,19H,4,18H2,1-3H3,(H,31,32)(H,33,37)(H,34,35,36)

2-(3-(4-((4-(1H-pyrazol-4-yl)phenyl)amino)-6-ethoxyquinazolin-2-yl)phenoxy)-N-isopropylacetamide

KL-11743; KL 11743; KL-11743; 1369452-53-8; 2-(3-(4-((4-(1H-pyrazol-4-yl)phenyl)amino)-6-ethoxyquinazolin-2-yl)phenoxy)-N-isopropylacetamide; NSC783733; CHEMBL4648818; SCHEMBL21681302; KL11743

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~25 mg/mL (~47.84 mM)

Solubility in Formulation 1: ≥ 2.08 mg/mL (3.98 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 900 μL of corn oil and mix evenly.

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 (Please use freshly prepared in vivo formulations for optimal results.)