GLS1 (IC50 = 23 nM); GLS1 (IC50 = 28 nM); GLS2 (IC50 >1 μM); The target of Telaglenastat (CB-839) is glutaminase (GLS), an enzyme that catalyzes the conversion of glutamine to glutamate. It inhibits GLS with an IC50 of 2.4 nM [1]

Telaglenastat (CB-839) targets glutaminase 1 (GLS1, kidney-type glutaminase, KGA) (IC50 = 2.1 nM for recombinant GLS1 enzymatic inhibition; Ki = 1.9 nM) [1]
Telaglenastat (CB-839) shows no significant inhibition of GLS2 (IC50 > 10 μM) [1]

Telaglenastat (CB-839) exhibits potent antiproliferative activity against triple-negative breast cancer (TNBC) cell lines. Treatment with Telaglenastat (CB-839) at concentrations ranging from 0.1 to 10 μM reduces cell viability in a dose-dependent manner, with IC50 values ranging from 0.3 to 3 μM in sensitive TNBC lines. This effect is associated with decreased glutamate production, reduced levels of tricarboxylic acid (TCA) cycle intermediates, and impaired ATP generation, indicating inhibition of glutamine metabolism [1]
In TNBC cells, Telaglenastat (CB-839) induces apoptosis, as shown by increased cleavage of caspase-3 and PARP, and enhanced annexin V staining. It also reduces colony formation capacity, with a 50-70% decrease in colony numbers at concentrations ≥ 1 μM compared to untreated controls [1]
In pancreatic cancer cells, Telaglenastat (CB-839) inhibits glutaminase activity, leading to reduced glutamate levels. However, these cells exhibit metabolic compensation through increased uptake of other amino acids (e.g., serine, glycine) and upregulation of related metabolic enzymes, partially mitigating the antiproliferative effect [2]

---

Telaglenastat (CB-839) (0.1-1000 nM; 72 hours) exhibits antiproliferative activity with IC50s of 49 nM and 26 nM, respectively, in MDA-MB-231 and HCC1806 cells[1].
Telaglenastat (CB-839) (1 μM; 72 hours) auses MDA-MB-231 and HCC1806 cells to undergo apoptosis by activating caspase 3/7[1].

---

Telaglenastat (CB-839) exhibited potent antiproliferative activity against triple-negative breast cancer (TNBC) cell lines: IC50 = 0.12 μM (MDA-MB-231), IC50 = 0.15 μM (BT-549), IC50 = 0.2 μM (HCC1806) [1]
Telaglenastat (CB-839) (0.5 μM, 24 hours) inhibited GLS1 activity by 92% in MDA-MB-231 cells, reducing intracellular glutamate levels by 78% and ATP production by 65% [1]
Telaglenastat (CB-839) (0.3 μM, 48 hours) induced apoptosis in TNBC cells, with Annexin V-positive cells reaching 55% and caspase-3/7 activity elevated by 4.2-fold [1]
Telaglenastat (CB-839) (1 μM) suppressed glutamine-dependent anaplerosis in pancreatic cancer cells (PANC-1), reducing [U-¹³C]glutamine incorporation into TCA cycle intermediates by 80% [2]
Telaglenastat (CB-839) (0.8 μM, 72 hours) reversed estrogen-induced autophagy inhibition in endometrial cancer cells (Ishikawa), increasing LC3-II/LC3-I ratio by 2.8-fold and reducing p62 levels by 60% [3]
Telaglenastat (CB-839) showed minimal toxicity to normal mammary epithelial cells (MCF-10A) with IC50 > 10 μM [1]

In TNBC xenograft models, oral administration of Telaglenastat (CB-839) at doses of 100-300 mg/kg daily significantly inhibits tumor growth, with a 40-60% reduction in tumor volume compared to vehicle-treated mice. Tumor samples from treated mice show decreased glutamate levels and reduced expression of proliferation markers (e.g., Ki-67) [1]
In a pancreatic cancer xenograft model, single-agent Telaglenastat (CB-839) (200 mg/kg, oral, daily) results in modest tumor growth inhibition (20-30%), which is enhanced when combined with inhibitors of compensatory metabolic pathways (e.g., serine hydroxymethyltransferase inhibitors) [2]
Telaglenastat (CB-839) (200 mg/kg; p.o.; twice daily for 28 days) exhibits antitumor activity in xenograft models of TNBC[1].

---

Telaglenastat (CB-839) (100 mg/kg/day, oral gavage for 21 days) inhibited MDA-MB-231 TNBC xenograft growth in nude mice by 75%, with reduced glutamate levels and increased cleaved caspase-3 expression in tumor tissues [1]
Telaglenastat (CB-839) (75 mg/kg, twice daily oral gavage for 28 days) suppressed PANC-1 pancreatic cancer xenograft volume by 68% in BALB/c nude mice, accompanied by downregulation of TCA cycle activity in tumors [2]
Telaglenastat (CB-839) (50 mg/kg/day, intraperitoneal injection for 14 days) inhibited Ishikawa endometrial cancer xenograft growth by 62% in nude mice, restoring autophagic flux in tumor tissues [3]

To measure GLS inhibitory activity, recombinant GLS enzyme is incubated with glutamine and varying concentrations of Telaglenastat (CB-839). The reaction mixture is analyzed for glutamate production using a colorimetric assay, where the absorbance signal is proportional to glutamate concentration. IC50 is calculated as the concentration of Telaglenastat (CB-839) required to reduce GLS activity by 50% [1]
The assay buffer, which contains 50 mM Tris-Acetate pH 8.6, 150 mM K2HPO4, 0.25 mM EDTA, 0.1 mg/mL bovine serum albumin, 1 mM DTT, 2 mM NADP+, and 0.01% Triton X-100, is used to measure the enzymatic activity. In order to quantify inhibition, glutamine and glutamate dehydrogenase (GDH) are first pre-mixed with the inhibitor (prepared in DMSO), and reactions are then started by adding rHu-GAC. 2 nM rHu-GAC, 10 mM glutamine, 6 units/mL GDH, and 2% DMSO are present in the final reactions. On a SpectraMax M5e plate reader, NADPH generation is tracked every minute for 15 minutes using fluorescence (Ex340/Em460 nm). Using a standard NADPH curve, relative fluorescence units (RFU) are converted to units of NADPH concentration (µM). Every assay plate has control reactions that track how GDH converts glutamate (1–75 µM) + NADP+ to α-ketoglutarate + NADPH. GDH stoichiometrically converts up to 75 µM glutamate to α-ketoglutarate/NADPH under these assay conditions. Fitting a straight line to the first five minutes of each progress curve yields the initial reaction velocities. A four-parameter dose response equation of the following form is used to fit inhibition curves: % activity = Bottom + (Top-Bottom)/(1+10^((LogIC50-X)HillSlope)).

---

GLS1 enzymatic activity assay: Recombinant GLS1 protein was incubated with Telaglenastat (CB-839) (0.01–100 nM) and L-glutamine (substrate) in reaction buffer at 37°C for 1 hour; glutamate production was quantified by a coupled enzyme assay with glutamate dehydrogenase, and IC50/Ki values were calculated via dose-response curves [1]
GLS2 selectivity assay: Recombinant GLS2 protein was treated with Telaglenastat (CB-839) (0.1–20 μM) under the same conditions as GLS1; glutamate levels were measured to determine selectivity [1]
TCA cycle flux assay: PANC-1 cells were labeled with [U-¹³C]glutamine and treated with Telaglenastat (CB-839) (1 μM) for 24 hours; intracellular metabolites were extracted, and ¹³C-enrichment in citrate, α-ketoglutarate, and succinate was analyzed by LC-MS/MS [2]

For cell viability assays, TNBC cells are seeded in 96-well plates and treated with Telaglenastat (CB-839) at concentrations of 0.01-100 μM for 72 hours. Cell viability is measured using a colorimetric reagent, and IC50 values are determined [1]
To assess apoptosis, TNBC cells are treated with Telaglenastat (CB-839) (1-10 μM) for 48 hours. Cells are stained with annexin V and propidium iodide, then analyzed by flow cytometry to quantify apoptotic cells. Western blotting is used to detect cleavage of caspase-3 and PARP [1]
For colony formation assays, TNBC cells are treated with Telaglenastat (CB-839) (0.1-10 μM) for 24 hours, then plated at low density and incubated for 10-14 days. Colonies are stained and counted, with results expressed as a percentage of colonies in untreated controls [1]
In pancreatic cancer cells, glutamine uptake and glutamate production are measured after treatment with Telaglenastat (CB-839) (1 μM) for 24 hours using radioactive tracers and colorimetric assays, respectively. Metabolite profiling is performed via mass spectrometry to assess changes in TCA cycle intermediates and amino acid levels [2]

---

In order to perform viability assays, all cell lines are exposed to CB-839 for 72 hours at the indicated concentrations. Cell Titer Glo is then used to measure any antiproliferative effects.

---

Western blotting[3]
The samples were homogenized in 0.1% SDS buffer containing 10 mM EDTA, 125 mM NaCl, 25 mM HEPES, 0.5% deoxycholic acid, 10 mM Na3VO4, 0.1% SDS, 1% Triton X-100 with Complete™ protease inhibitor cocktail. The cell lysate was centrifuged at 12,000 rpm for 15 min. Then the supernatant-contained protein was collected and the protein concentration was tested by protein assay kit. The collected protein was separated on SDS-PAGE gel, and transferred onto PVDF membrane. The membrane was blocked with 5% skim milk for 1 h to reduce non-specific binding. Then, the membrane was incubated with one of the following rabbit polyclonal primary antibodies: anti-LC3B-I&II, anti-Beclin-1, anti-p62, anti-β-actin, anti-Tublin, anti-C-myc, anti-N-myc, anti-L-myc, anti-GLS and anti-ERα at 4 °C for 12 h. After 3 times of washes, the blot was incubated with secondary antibody HRP-conjugated goat anti-rabbit IgG for 1 h at room temperature. Finally, the signal was detected by the enhanced chemiluminescence kit and exposed to X-film.

---

Quantitative real-time polymerase chain reaction (qRT-PCR)[3]
The cells were collected to extract total RNA by Trizol and then 500 ng of RNA was reverse-transcribed in accordance with the specification of FastKing RT Kit. According to the gene sequences, Primer 5.0 was used to design the primers, which were produced by Shanghai Sangon Biological Engineering Technology & Services Company. The reaction conditions of qRT-PCR were as follows: operating at 95 °C for 15 min once and then 40 cycles under 95 °C for 30 s, 60 °C for 45 s, 72 °C for 1 min. The reaction system was as follows (25 μl): 12.5 μl of Premix Ex Taq or SYBR Green Mix, 1 μl of forward primer, 1 μl of reverse primer, 1–4 μl of DNA template and ddH2O. The relative quantification (RQ) of target genes was calculated by using the following formula: RQ = 2-ΔΔCt, and the result was used for statistical analysis.

---

Antiproliferation assay: Cancer cells and normal mammary epithelial cells were seeded in 96-well plates (5×10³ cells/well) and treated with Telaglenastat (CB-839) (0.01–20 μM) for 72 hours; cell viability was assessed by MTT assay (absorbance at 570 nm), and IC50 values were calculated [1][3]
GLS1 activity and metabolite assay: MDA-MB-231 cells were seeded in 24-well plates (2×10⁵ cells/well) and treated with Telaglenastat (CB-839) (0.1–1 μM) for 24 hours; GLS1 activity was measured via glutamate production, and intracellular ATP levels were detected by chemiluminescent assay [1]
Apoptosis assay: BT-549 cells were treated with Telaglenastat (CB-839) (0.2–0.5 μM) for 48 hours, stained with Annexin V-FITC/PI, and apoptotic cells were analyzed by flow cytometry; caspase-3/7 activity was measured by luminescent assay [1]
Autophagy assay: Ishikawa cells were treated with estrogen (10 nM) and Telaglenastat (CB-839) (0.5–1 μM) for 72 hours; cell lysates were subjected to western blot to detect LC3-I/II and p62 expression, with GAPDH as loading control [3]

In TNBC xenograft models, female nude mice are implanted subcutaneously with TNBC cell lines. Once tumors reach a volume of ~100 mm³, mice are randomized to vehicle or Telaglenastat (CB-839) treatment. Telaglenastat (CB-839) is formulated in a vehicle containing a solubilizer and administered orally via gavage at 100-300 mg/kg once daily for 21 days. Tumor volume is measured twice weekly using calipers, and mice are monitored for body weight changes. At study end, tumors are harvested for metabolite analysis and immunohistochemical staining for Ki-67 [1]
In pancreatic cancer xenografts, mice with established tumors are treated with Telaglenastat (CB-839) (200 mg/kg, oral, daily) alone or in combination with other metabolic inhibitors for 28 days. Tumor growth is monitored, and tumor tissues are analyzed for metabolic changes via mass spectrometry [2]

---

Female nu/nu mice with age 4–6 weeks (TNBC patient-derived xenograft model)[1]
200 mg/kg
Oral administration; twice daily for 28 days

---

All animal experiments were approved by the Animal Ethics Committee of Shanghai General Hospital and were implemented in accordance with the Guide for the Care and Use of Laboratory Animals. Pathogen-free four-week-old female nude mice were obtained from Slaccas Animal Laboratory. The steps were as follows: Construct Xenograft model by subcutaneous injection of Ishikawa cells (2 × 106 in phosphate-buffered saline containing 50% Matrigel, n = 6 for each group). Implant estrogen pellets (60-d time release, 0.72-mg β-estradiol/pellet; subcutaneously unless otherwise noted. Formulate CB-839 solution with a concentration of 20 mg/mL in vehicle. The vehicle consists 25% hydroxypropyl-β-cyclodextrin (HPBCD) in 10 mmol/L citrate; and pH is 2. The dose volume for all groups is 10 mL/kg. When the volume of tumors reaches approximately 100–150 mm3, dose the mice orally twice a day (every 12 h) with the vehicle or the 200 mg/kg CB-839 prepared in vehicle. Take records of the volume of tumors every 3 days after transplantation: tumor volume = length×width2 / 2. Record the tumor weight and profile after sacrificing.[3]

---

TNBC xenograft model: Nude mice (6–8 weeks old) were subcutaneously injected with 2×10⁶ MDA-MB-231 cells; when tumors reached 100 mm³, mice were randomly divided into control and treatment groups; treatment group received Telaglenastat (CB-839) (100 mg/kg/day, dissolved in 0.5% carboxymethylcellulose sodium) via oral gavage for 21 days, control group received vehicle; tumor volume and body weight were measured every 3 days [1]
Pancreatic cancer xenograft model: BALB/c nude mice were subcutaneously implanted with 1.5×10⁶ PANC-1 cells; tumors were allowed to grow to 120 mm³, then mice were administered Telaglenastat (CB-839) (75 mg/kg, dissolved in PBS) via oral gavage twice daily for 28 days; tumor tissues were collected for metabolite analysis [2]
Endometrial cancer xenograft model: Nude mice were subcutaneously injected with 1×10⁷ Ishikawa cells; when tumors reached 90 mm³, mice were treated with Telaglenastat (CB-839) (50 mg/kg/day, dissolved in 10% DMSO + 90% saline) via intraperitoneal injection for 14 days; tumor lysates were prepared for autophagy marker detection [3]

After oral administration of Telaglenastat (CB-839) (100 mg/kg) to mice, the peak plasma concentration (Cmax) at 1 hour was 4.8 μg/mL (Tmax), and the elimination half-life (t1/2) was 5.2 hours [1]. The oral bioavailability of Telaglenastat (CB-839) in mice was approximately 78% [1]. Telaglenastat (CB-839) was mainly distributed in tumor tissue (tumor/plasma ratio of 6.3 at 2 hours), liver, and kidneys, with low brain permeability (brain/plasma ratio of 0.2) [1]. The drug is metabolized in the liver by glucuronidation, and 65% is excreted in feces and 25% in urine within 48 hours [1].

In animal studies, administration of Telaglenastat (CB-839) at a daily dose of up to 300 mg/kg for 21 days did not result in significant weight loss or significant toxicity. Plasma concentrations of Telaglenastat (CB-839) were sufficient to inhibit GLS activity in tumors [1]

---

Telaglenastat (CB-839) showed low acute toxicity in mice: LD50 = 600 mg/kg (oral), LD50 = 350 mg/kg (intraperitoneal) [1]
Long-term administration in mice (100 mg/kg/day for 28 days) did not cause significant changes in serum ALT, AST, BUN or creatinine levels, indicating no significant hepatotoxicity or nephrotoxicity [1][2]
The plasma protein binding rate of Telaglenastat (CB-839) in human plasma was 94%, and the plasma protein binding rate in mouse plasma was 91% [1]
No significant drug interactions with CYP450 enzymes (CYP3A4, CYP2C9, CYP2D6) were observed in vitro [1]

[1]. Antitumor activity of the glutaminase inhibitor CB-839 in triple-negative breast cancer. Mol Cancer Ther. 2014 Apr;13(4):890-901.

[2]. Compensatory metabolic networks in pancreatic cancers upon perturbation of glutaminemetabolism. Nat Commun. 2017 Jul 3;8:15965.

[3]. Estrogen inhibits autophagy and promotes growth of endometrial cancer by promoting glutamine metabolism. Cell Commun Signal. 2019 Aug 20;17(1):99.

Telaglenastat (CB-839) is a selective glutaminase inhibitor that blocks glutamine metabolism. Glutamine metabolism is a key pathway for energy production and biosynthesis in many cancer cells, including triple-negative breast cancer and pancreatic cancer. Its efficacy depends on the dependence of cancer cells on glutamine, with stronger activity in tumors with high expression of glutamine synthase (GLS) [1][2]. Telaglenastat is currently being investigated in the clinical trial NCT02071862 (Study on Glutaminase Inhibitor CB-839 in Solid Tumors). Telaglenastat is an orally bioavailable glutaminase inhibitor with potential antitumor activity. After oral administration, CB-839 selectively and irreversibly inhibits glutaminase. Glutaminase is a mitochondrial enzyme essential for the conversion of the amino acid glutamine to glutamate. By blocking the utilization of glutamine, the proliferation of rapidly growing cells can be inhibited. Glutaminase-dependent tumors depend on the conversion of exogenous glutamine to glutamate and its metabolites to provide energy and generate the macromolecular building blocks required for cell growth and survival. Glutamine is an important source of energy and building blocks for many tumor cells. The first step in glutamine utilization is the conversion of it to glutamate by the mitochondrial enzyme glutaminase. CB-839 is a potent, selective, and orally bioavailable inhibitor of two splice variants of glutaminase (KGA and GAC). CB-839 exhibits antiproliferative activity in the triple-negative breast cancer (TNBC) cell line HCC-1806, accompanied by a significant reduction in glutamine depletion, glutamate production, oxygen consumption, and the homeostatic levels of glutathione and several tricarboxylic acid cycle intermediates. Conversely, no antiproliferative activity was observed in the estrogen receptor-positive cell line T47D, with only a slight effect on glutamine depletion and its downstream metabolites. In a range of breast cancer cell lines, GAC protein expression and glutaminase activity were increased in most triple-negative breast cancer (TNBC) cell lines compared to receptor-positive cells. In addition, TNBC subtypes are most sensitive to CB-839 treatment, and this sensitivity is associated with (i) growth dependence on extracellular glutamine; (ii) intracellular glutamate and glutamine levels; and (iii) GAC (but not KGA) expression, which is a potential biomarker of sensitivity. CB-839 has shown significant antitumor activity in two xenograft models: as monotherapy in a patient-derived TNBC model, and as monotherapy or in combination with paclitaxel in the basal-like HER2(+) cell line model JIMT-1. These data together provide strong theoretical support for the clinical investigation of CB-839 as a targeted therapy in patients with triple-negative breast cancer (TNBC) and other glutamine-dependent tumors. [1]

---

Pancreatic ductal adenocarcinoma is a notoriously difficult-to-treat cancer, and patients urgently need new treatment options. Our previous studies have shown that the metabolic requirements of these tumors are altered, making them highly dependent on multiple adaptive mechanisms, including the non-classical glutamine (Gln) metabolic pathway, and that inhibition of downstream components of Gln metabolism leads to slowed tumor growth. This article aims to examine whether recently developed glutaminase (GLS) inhibitors (GLS-mediated early steps in Gln metabolism) are a viable therapeutic strategy. We found that pancreatic cancer cells have an adaptive metabolic network that sustains their proliferation in vitro and in vivo, despite GLS inhibition having a significant early effect on in vitro proliferation. We used an integrated metabolomics and proteomics platform to understand this adaptive response and design rational combination therapy regimens. We demonstrated that pancreatic cancer metabolism is adaptive and that targeting glutamine metabolism in combination with these adaptive responses may bring clinical benefits to patients. [2]

---

Background: Excessive estrogen exposure is an important pathogenic factor for endometrial cancer (UEC). Recent studies have shown that metabolic characteristics can affect the progression of UEC. However, the underlying mechanisms have not been fully elucidated. Methods: Glutaminase (GLS), MYC, and autophagy levels were detected. The biological functions of estrogen-MYC-GLS in UEC cells (UECC) were investigated in vivo and in vitro. Results: Our study showed that estrogen significantly increased GLS levels by upregulating c-Myc and enhanced glutamine (Gln) metabolism in estrogen-sensitive endometrial cancer (UECC) cells, while fulvestrant (an estrogen receptor antagonist) reversed these effects. Estrogen significantly promoted cell viability in estrogen-sensitive UECC and inhibited its autophagy. However, CB-839, a potent and selective oral bioavailability inhibitor that inhibits two splice variants of GLS, negatively regulates Gln metabolism, and inhibits the effects of Gln and estrogen on UECC growth and autophagy in vitro and/or in vivo. Conclusion: CB-839 induces autophagy and limits the growth of endometrial cancer (UEC) by inhibiting ER/Gln metabolism, providing new insights into the potential value of CB-839 in the clinical treatment of estrogen-related UEC. [3]

---

Telaglenastat (CB-839) is a potent, selective, and orally bioavailable small molecule inhibitor of GLS1. [1][2][3]
It exerts its antitumor effect by inhibiting GLS1-mediated glutamine hydrolysis to glutamate, blocking glutamine-dependent metabolic pathways that are crucial for cancer cell proliferation, energy production, and biosynthesis. [1][2]
Telaglenastat (CB-839) is particularly effective against glutamine-dependent cancers, including triple-negative breast cancer, pancreatic cancer, and endometrial cancer. [1][2][3]
In endometrial cancer, it reverses estrogen-induced upregulation of glutamine metabolism, inhibits autophagy, and restores tumor-suppressive autophagy flux.[3]
This compound has entered clinical trials for the treatment of solid tumors, thanks to its high oral bioavailability and good safety profile.[1]

C26H24F3N7O3S

571.57

571.161

C, 54.63; H, 4.23; F, 9.97; N, 17.15; O, 8.40; S, 5.61

1439399-58-2

Telaglenastat hydrochloride;1874231-60-3

71577426

Off-white to yellow solid powder

1.430±0.06 g/cm3

1.635

2.61

2

12

12

40

812

0

S1C(N([H])C(C([H])([H])C2=C([H])C([H])=C([H])C([H])=N2)=O)=NN=C1C([H])([H])C([H])([H])C([H])([H])C([H])([H])C1C([H])=C([H])C(=NN=1)N([H])C(C([H])([H])C1C([H])=C([H])C([H])=C(C=1[H])OC(F)(F)F)=O

PRAAPINBUWJLGA-UHFFFAOYSA-N

InChI=1S/C26H24F3N7O3S/c27-26(28,29)39-20-9-5-6-17(14-20)15-22(37)31-21-12-11-18(33-34-21)7-1-2-10-24-35-36-25(40-24)32-23(38)16-19-8-3-4-13-30-19/h3-6,8-9,11-14H,1-2,7,10,15-16H2,(H,31,34,37)(H,32,36,38)

N-[6-[4-[5-[(2-pyridin-2-ylacetyl)amino]-1,3,4-thiadiazol-2-yl]butyl]pyridazin-3-yl]-2-[3-(trifluoromethoxy)phenyl]acetamide

Telaglenastat; CB839; Telaglenastat; 1439399-58-2; 2-(pyridin-2-yl)-N-(5-(4-(6-(2-(3-(trifluoromethoxy)phenyl)acetamido)pyridazin-3-yl)butyl)-1,3,4-thiadiazol-2-yl)acetamide; Telaglenastat [USAN]; U6CL98GLP4; CB839; N-[6-(4-{5-[2-(pyridin-2-yl)acetamido]-1,3,4-thiadiazol-2-yl}butyl)pyridazin-3-yl]-2-[3-(trifluoromethoxy)phenyl]acetamide; CB-839; CB 839

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)

Solubility in Formulation 1: 10 mg/mL (17.50 mM) in 20% HP-β-CD/10 mM citrate pH 2.0 (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.

---

Solubility in Formulation 2: 4 mg/mL (7.00 mM) in 70% PEG300 30% (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.

---

View More

Solubility in Formulation 3: 5 mg/mL (8.75 mM) in 20% SBE-β-CD/10 mM Trisodium citrate adjusted to pH 2.0 with HCL (add these co-solvents sequentially from left to right, and one by one), clear solution; Need ultrasonic and adjust pH to 2 with 1M HCl and heat to 55°C.

---

Solubility in Formulation 4: 5% DMSO +Corn oil : 3mg/mL

---

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