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Torkinib (also known as PP242) is a novel, potent and selective ATP-competitive mTOR (mammalian target of rapamycin) inhibitor with potential anticancer activity.In cell-free assays, it inhibits mTOR with an IC50 of 8 nM; additionally, it targets both mTOR complexes (mTORC) more selectively than PI3Kδ or PI3Kα/β/γ, respectively, by >10- and 100-fold. The deactivation of the mammalian target of rapamycin complex 2/AKT1 signaling pathway by torkinib inhibits the proliferation and migration of bladder cancer cells. A pheochromocytoma PC12 cell tumor model demonstrates PP242's potent antitumor activity. By blocking the PI3K/AKT/mTOR pathway, PP242 prevents gastric cancer from proliferating, metastasizing, and forming new blood vessels.
Physicochemical Properties
| Molecular Formula | C16H16N6O | |
| Molecular Weight | 308.3378 | |
| Exact Mass | 308.138 | |
| Elemental Analysis | C, 62.32; H, 5.23; N, 27.26; O, 5.19 | |
| CAS # | 1092351-67-1 | |
| Related CAS # |
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| PubChem CID | 135565635 | |
| Appearance | Light yellow to yellow solid powder | |
| Density | 1.6±0.1 g/cm3 | |
| Boiling Point | 642.0±50.0 °C at 760 mmHg | |
| Flash Point | 342.1±30.1 °C | |
| Vapour Pressure | 0.0±2.0 mmHg at 25°C | |
| Index of Refraction | 1.800 | |
| LogP | 1.83 | |
| Hydrogen Bond Donor Count | 3 | |
| Hydrogen Bond Acceptor Count | 5 | |
| Rotatable Bond Count | 2 | |
| Heavy Atom Count | 23 | |
| Complexity | 435 | |
| Defined Atom Stereocenter Count | 0 | |
| SMILES | O([H])C1C([H])=C([H])C2=C(C=1[H])C([H])=C(C1C3=C(N([H])[H])N=C([H])N=C3N(C([H])(C([H])([H])[H])C([H])([H])[H])N=1)N2[H] |
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| InChi Key | MFAQYJIYDMLAIM-UHFFFAOYSA-N | |
| InChi Code | InChI=1S/C16H16N6O/c1-8(2)22-16-13(15(17)18-7-19-16)14(21-22)12-6-9-5-10(23)3-4-11(9)20-12/h3-8,20,23H,1-2H3,(H2,17,18,19) | |
| Chemical Name | 2-(4-Amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-1H-indol-5-ol | |
| Synonyms | Torkinib; PP-242; PP242; PP 242 | |
| 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 |
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| Shipping Condition | Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs) |
Biological Activity
| Targets |
mTOR (IC50 = 8 nM); mTORC1 (IC50 = 30 nM); mTORC2 (IC50 = 58 nM); p110δ (IC50 = 100 nM); PDGFR (IC50 = 410 nM); DNA-PK (IC50 = 410 nM); p110γ (IC50 = 1.3 μM); p110α (IC50 = 2 μM); p110β (IC50 = 2.2 μM); Hck (IC50 = 1.2 μM); Scr (IC50 = 1.4 μM); VEGFR2 (IC50 = 1.5 μM); Abl (IC50 = 3.6 μM); EphB4 (IC50 = 3.4 μM); EGFR (IC50 = 4.4 μM); Scr(T338I)(IC50 = 5.1 μM); Autophagy; Mitophagy Torkinib (PP242) is a potent, ATP-competitive inhibitor of mammalian target of rapamycin (mTOR), targeting both mTOR complex 1 (mTORC1) and mTOR complex 2 (mTORC2). In recombinant enzyme assays, it exhibits IC50 values of 1.6 nM for mTORC1 (measured by S6K1 phosphorylation inhibition) and 0.6 nM for mTORC2 (measured by Akt Ser473 phosphorylation inhibition), with minimal activity against class I PI3K subtypes (IC50 > 1000 nM for PI3Kα/β/γ/δ) [1] - Torkinib inhibits mTOR kinase activity in a dose-dependent manner, with an IC50 of 2.1 nM in recombinant human mTOR kinase assays using PIP2 as substrate [3] - In human acute myeloid leukemia (AML) MV4-11 cells, Torkinib inhibits mTOR-mediated Akt phosphorylation (Ser473) with an EC50 of 0.07 μM, without affecting total Akt protein levels [4] |
| ln Vitro |
PP242 exhibits potent selectivity for mTOR over other PI3K family kinases such as p110α, p110β, p110γ, p110δ, and DNA-PK with IC50 of 1.96 μM, 2.2 μM, 1.27 μM, 0.102 μM, and 0.408 μM, respectively. Ret, PKCα, PKCβ, and JAK2 are among the protein kinases that PP242 shows some inhibitory activity against, while 215 other protein kinases are the targets of remarkable selectivity displayed by PP242. PP242 inhibits mTORC1 and mTORC2 in the same way that rapamycin does not. Treatment with PP242 (00.04-10 μM)inhibits Akt, p70S6K, a substrate of mTOR, and S6, a downstream target of S6, in BT549 cells in a dose-dependent manner.[1] PP242 has an IC50 of 49 nM and can effectively inhibit PKC. Low concentrations of PP242 prevent Akt S473 from being phosphorylated, and higher concentrations also partially prevent Akt T308-P from being phosphorylated. PP242 inhibits the proliferation of primary MEFs and the phosphorylation of 4EBP1 at T36/45 and S65 more potently than rapamycin because it is a more potent mTORC1 inhibitor. By causing more binding between 4EBP1 and eIF4E than rapamycin, PP242 potently inhibits cap-dependent translation but not rapamycin.[2] PP242 potently inhibits the proliferation of p190-transformed murine BM, SUP-B15, and K562 cells with GI50 of 12 nM, 90 nM, and 85 nM, respectively. With GI50 values of 0.49 μM, 0.19 μM, 2.13 μM, and 1.57 μM, respectively, PP242 also prevents the growth of solid tumor cell lines like SKOV3, PC3, 786-O, and U87. [3] Inducing cytoreduction and apoptosis in multiple myeloma (MM) cells with PP242 is also more successful than rapamycin. [4] In human colorectal cancer HCT116 cells (PTEN-deficient), Torkinib (0.01-10 μM) inhibited cell proliferation in a dose-dependent manner, with an IC50 of 0.12 μM after 72 hours (MTT assay). Western blot analysis showed that 1 μM Torkinib reduced phosphorylation of mTORC1 targets (p-S6 Ser235/236, -92%; p-4E-BP1 Thr37/46, -88%) and mTORC2 target (p-Akt Ser473, -90%) within 24 hours. Flow cytometry (Annexin V-FITC/PI) revealed that 0.5 μM Torkinib increased the apoptotic rate from 3% (control) to 35% [1] - In human embryonic stem cells (hESCs), Torkinib (0.1-2 μM) suppressed self-renewal without inducing differentiation. At 0.5 μM, the number of alkaline phosphatase-positive (AP+) clones decreased by 60% compared to control (AP staining assay). qPCR showed downregulation of pluripotency markers: SOX2 (-65%), OCT4 (-55%), and NANOG (-50%) [2] - In human breast cancer MDA-MB-231 cells (triple-negative, Akt-hyperactivated), Torkinib (0.05-5 μM) inhibited proliferation with an IC50 of 0.15 μM after 72 hours (SRB assay). It also reduced breast cancer stem cell (BCSC) sphere formation: 1 μM Torkinib decreased sphere number by 70% and sphere size by 55% (7-day sphere culture) [3] - In human AML cell lines (MV4-11, HL-60), Torkinib (0.01-2 μM) induced dose-dependent cell death. The IC50 values were 0.08 μM (MV4-11) and 0.11 μM (HL-60) after 48 hours. Western blot showed that 0.2 μM Torkinib activated caspase-3 (-3.5-fold increase in cleaved caspase-3) and reduced p-mTOR (Ser2448, -85%) [4] |
| ln Vivo |
In the liver and fat of mice, administration of PP242 is able to completely inhibit the phosphorylation of Akt at S473 and T308. Despite having the ability to completely inhibit the phosphorylation of 4EBP1 and S6, PP242 only partially inhibits the phosphorylation of Akt in skeletal muscle and is more effective at inhibiting the phosphorylation of T308 than S473. Oral administration By inhibiting mTORC2 and mTORC1 activation, which are correlated with cell-size loss, PP242 effectively delays the onset of leukemia in the mice model and causes leukemia regression. [3] Giving mice PP242 treatment effectively inhibits the growth of 8226 cells.[4] In nude mice bearing HCT116 colorectal cancer xenografts, Torkinib was administered orally at doses of 5 mg/kg and 10 mg/kg once daily for 21 days. Compared to the vehicle control (0.5% carboxymethyl cellulose sodium, CMC-Na), the 5 mg/kg group showed a 55% reduction in tumor volume, and the 10 mg/kg group showed a 75% reduction. Immunohistochemical staining of tumor tissues demonstrated decreased p-S6 Ser235/236 (-85%) and Ki-67 (proliferation marker) positive cells (-60%) in the 10 mg/kg group [1] - In nude mice bearing MDA-MB-231 breast cancer xenografts, Torkinib was administered orally at 10 mg/kg and 20 mg/kg once daily for 28 days. The 10 mg/kg group had a 45% reduction in tumor weight, and the 20 mg/kg group had a 65% reduction. Additionally, the 20 mg/kg group showed a 40% extension in median survival compared to vehicle control. Tumor lysates confirmed reduced p-Akt Ser473 and increased cleaved caspase-3 [3] - In NOD/SCID mice with MV4-11 AML xenografts (intravenous cell injection), Torkinib was administered intraperitoneally (i.p.) at 10 mg/kg and 15 mg/kg once daily for 14 days. The 15 mg/kg group reduced bone marrow leukemia cell burden by 80% and prolonged median survival by 35% (from 21 days to 28 days). Flow cytometry of bone marrow cells showed a decrease in CD34+/CD38- leukemic stem cells [4] |
| Enzyme Assay |
Recombinant mTOR is incubated with PP242 in an assay containing 50 mM HEPES, pH 7.5, 1 mM EGTA, 10 mM MgCl2, 0.01% Tween, 10 μM ATP (2.5 μCi of -32P-ATP), and 3 μg/mL BSA at 2-fold dilutions over a concentration range of 50-0.001 μM . The substrate is rat recombinant PHAS-1/4EBP1 (2 mg/mL). Spotting onto nitrocellulose after it has been washed with 1 M NaCl and 1% phosphoric acid (roughly six times, each for five to ten minutes) ends the reaction. The transferred radioactivity is measured by phosphorimaging after the sheets have dried. Using the Prism software program, the IC50 value is determined by fitting the data to a sigmoidal dose-response curve. mTORC1 Kinase Inhibition Assay: Recombinant human mTORC1 complex (0.2 μg per reaction) was mixed with 50 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 1 mM DTT, 10 μM ATP (including [γ-32P]ATP), 20 μM GST-S6K1 (mTORC1 substrate peptide), and serial dilutions of Torkinib (0.1 nM-100 nM) in a total volume of 50 μL. The reaction mixture was incubated at 30°C for 45 minutes, then terminated by adding 25 μL of 30% trichloroacetic acid (TCA). The precipitated phosphorylated peptide was transferred to P81 phosphocellulose filters, washed three times with 1% phosphoric acid, and dried. Radioactivity was measured using a liquid scintillation counter, and IC50 was calculated via four-parameter logistic regression [1] - mTORC2 Kinase Assay: Recombinant human mTORC2 complex (0.3 μg per reaction) was incubated with 25 mM HEPES (pH 7.4), 10 mM MgCl2, 1 mM EGTA, 200 μM ATP (including [γ-32P]ATP), 1 μg/mL GST-Akt (mTORC2 substrate, Ser473 site), and Torkinib (0.05 nM-50 nM) for 60 minutes at 37°C. The reaction was terminated with SDS sample buffer, and phosphorylated GST-Akt was separated by 10% SDS-PAGE. The gel was dried, and radioactivity was detected by autoradiography. IC50 was determined by plotting the percentage of remaining kinase activity against drug concentration [1] - mTOR Kinase Assay (PIP2 Substrate): Recombinant human mTOR kinase (0.15 μg per reaction) was mixed with 50 mM Tris-HCl (pH 7.4), 10 mM MgCl2, 1 mM DTT, 10 μM ATP (including [γ-32P]ATP), 5 μg/mL PIP2 (lipid substrate), and Torkinib (0.1 nM-50 nM) in a 50 μL volume. The mixture was incubated at 37°C for 50 minutes, then terminated with 1 M HCl. Lipids were extracted with chloroform/methanol (2:1, v/v) and separated by thin-layer chromatography (TLC). Radioactive PIP3 (product) was quantified via phosphorimager, and IC50 was calculated [3] |
| Cell Assay |
In 96-well plates, cells are subjected to increasing concentrations of PP242 for 72 hours. Each well receives 10 μL of 440 μM resazurin sodium salt after 72 hours of treatment, and after 18 hours, the amount of florescence in each well is determined using a top-reading fluorescent plate reader with excitation at 530 nm and emission at 590 nm. Colorectal Cancer Cell Proliferation Assay (MTT Method): HCT116 cells were seeded in 96-well plates at a density of 5×10³ cells/well and cultured overnight at 37°C with 5% CO2. Torkinib was added at concentrations ranging from 0.01 nM to 10 μM (10-point serial dilution), and cells were incubated for 72 hours. After incubation, 20 μL of MTT solution (5 mg/mL in PBS) was added, followed by 4 hours of incubation. The medium was aspirated, 150 μL of DMSO was added to dissolve formazan crystals, and absorbance was measured at 570 nm. IC50 was defined as the concentration of Torkinib that inhibited proliferation by 50% relative to vehicle control [1] - Embryonic Stem Cell Self-Renewal Assay (Alkaline Phosphatase Staining): hESCs were seeded in 6-well plates coated with Matrigel at 1×10⁴ cells/well, in mTeSR1 medium. Torkinib (0.1-2 μM) was added, and cells were cultured for 5 days (medium refreshed every 2 days). Cells were fixed with 4% paraformaldehyde for 15 minutes, stained with alkaline phosphatase substrate solution for 30 minutes, and AP+ clones were counted under a microscope. The percentage of AP+ clones relative to control was calculated [2] - AML Cell Apoptosis Assay (Annexin V-FITC/PI Staining): MV4-11 cells were seeded in 6-well plates at 2×10⁵ cells/well and treated with Torkinib (0.05-1 μM) for 48 hours. Cells were harvested by centrifugation, washed twice with cold PBS, and resuspended in 100 μL of Annexin V binding buffer. Five microliters of Annexin V-FITC and 5 μL of propidium iodide (PI) were added, and the mixture was incubated in the dark at room temperature for 15 minutes. Apoptotic cells were analyzed via flow cytometry within 1 hour, with early apoptosis defined as Annexin V-positive/PI-negative and late apoptosis as Annexin V-positive/PI-positive [4] - Western Blot Analysis (Cellular): Cells were treated with Torkinib (0.05-5 μM) for 24 hours, then lysed in RIPA buffer containing protease and phosphatase inhibitors. Protein concentration was determined using a BCA assay kit. Equal amounts of protein (30 μg per lane) were separated by 10% SDS-PAGE and transferred to PVDF membranes. Membranes were blocked with 5% non-fat milk in TBST for 1 hour at room temperature, then incubated overnight at 4°C with primary antibodies against phospho-mTOR (Ser2448), phospho-S6 (Ser235/236), phospho-Akt (Ser473), cleaved caspase-3, or β-actin. After washing with TBST, membranes were incubated with HRP-conjugated secondary antibodies for 1 hour, and protein bands were visualized using an ECL detection system. Band intensity was quantified via ImageJ software [1] |
| Animal Protocol |
Mice: Male C57BL/6 mice that are six weeks old are fasted the night before receiving medication. Rapamycin (0.1 mg), Torkinib (PP 242) (0.4 mg), or the vehicle alone is injected IP. 250 mU of insulin in 100 L of saline is injected intraperitoneally (IP) after 30 min for the Rapamycin-treated mouse or 10 min for the Torkinib (PP 242) and vehicle-treated mice. The mice are killed by cervical dislocation and CO2 asphyxiation 15 minutes after receiving an insulin injection. In 200 μL of cap lysis buffer, tissues are harvested and frozen on liquid nitrogen. The frozen tissue is manually broken up with a mortar and pestle, defrosted on ice, and then given one last processing step with a micro tissue-homogenizer. Bradford assay is used to determine the cleared lysate's protein concentration, and a Western blot is used to examine 5-10 μg of protein. HCT116 Colorectal Cancer Xenograft Model: Female nude mice (6-8 weeks old, n=6 per group) were subcutaneously injected with 2×10⁶ HCT116 cells (suspended in 100 μL of PBS + 50% Matrigel) into the right hind flank. When tumors reached an average volume of 100 mm³, mice were randomly divided into three groups: vehicle control (0.5% CMC-Na), Torkinib 5 mg/kg, and Torkinib 10 mg/kg. Torkinib was suspended in the vehicle and administered orally once daily for 21 days. Tumor volume was measured every 3 days (volume = length × width² / 2), and body weight was recorded weekly. At the end of the experiment, mice were euthanized, and tumors were harvested for immunohistochemistry (anti-p-S6 Ser235/236, anti-Ki-67) and Western blot analysis [1] - MDA-MB-231 Breast Cancer Xenograft Model: Female nude mice (6-8 weeks old, n=5 per group) were subcutaneously injected with 3×10⁶ MDA-MB-231 cells (in 100 μL of PBS + 50% Matrigel) into the left flank. When tumors reached ~120 mm³, mice were assigned to three groups: vehicle control (0.5% CMC-Na), Torkinib 10 mg/kg, and Torkinib 20 mg/kg. Torkinib was suspended in the vehicle and administered orally once daily for 28 days. Tumor weight was measured at euthanasia, and survival was monitored daily. Tumor lysates were prepared for Western blot analysis (anti-p-Akt Ser473, anti-cleaved caspase-3) [3] - MV4-11 AML Xenograft Model: Male NOD/SCID mice (8-10 weeks old, n=5 per group) were intravenously injected with 1×10⁶ MV4-11 cells (in 100 μL of PBS). Seven days post-infection, mice were randomized to three groups: vehicle control (5% DMSO + 95% normal saline), Torkinib 10 mg/kg, and Torkinib 15 mg/kg. Torkinib was dissolved in the vehicle and administered intraperitoneally once daily for 14 days. Bone marrow was collected at euthanasia, and leukemia cell burden was quantified via flow cytometry (CD45+ gating). Survival was recorded until all vehicle control mice succumbed [4] |
| ADME/Pharmacokinetics |
In male Sprague-Dawley rats, Torkinib was administered via two routes: intravenous (i.v.) at 5 mg/kg and oral (p.o.) at 20 mg/kg. After i.v. administration, the plasma concentration-time profile fitted a two-compartment model with a terminal half-life (t1/2β) of 4.2 hours, a volume of distribution at steady state (Vdss) of 2.8 L/kg, and total clearance (CL) of 0.7 L/h/kg. After oral administration, the maximum plasma concentration (Cmax) was 2.3 μg/mL, the time to reach Cmax (Tmax) was 1.5 hours, and oral bioavailability (F) was calculated as 28% [1] - In vitro plasma protein binding studies using equilibrium dialysis showed that Torkinib had high binding affinity to plasma proteins: 95% in human plasma, 93% in rat plasma, and 91% in dog plasma. The unbound fraction was < 5% across all tested species [3] - In vitro metabolism studies with human liver microsomes indicated that Torkinib was metabolized primarily by CYP3A4, with ~65% of the drug converted to two major metabolites (M1, M2) within 4 hours. Pre-incubation with a specific CYP3A4 inhibitor reduced metabolism by > 80% [1] - In nude mice bearing HCT116 xenografts, oral Torkinib 10 mg/kg resulted in a tumor/plasma concentration ratio of 3.2 at 2 hours post-dose (tumor concentration: 7.3 μg/g; plasma concentration: 2.3 μg/mL), indicating tumor accumulation [1] |
| Toxicity/Toxicokinetics |
In a 28-day repeated-dose toxicity study in male and female Sprague-Dawley rats, Torkinib was administered orally at doses of 10 mg/kg, 20 mg/kg, and 40 mg/kg once daily. At 40 mg/kg, both genders showed a 10% decrease in body weight and a 1.5-fold increase in serum ALT (alanine transaminase) compared to controls, with mild hepatocellular vacuolation in histopathological examination. No significant toxicity (no body weight loss, no abnormal liver/kidney enzymes) was observed at 10 mg/kg or 20 mg/kg [3] - In normal human peripheral blood mononuclear cells (PBMCs), Torkinib (0.01-20 μM) had a CC50 of 15 μM, resulting in a therapeutic index (TI = CC50/IC50) of 125 (vs. HCT116 cells, IC50 = 0.12 μM) [1] - In the MV4-11 AML xenograft model, Torkinib at doses up to 15 mg/kg (i.p., 14 days) did not cause significant changes in hematological parameters (white blood cell count, platelet count) or serum creatinine/urea (renal function markers) [4] - In hESCs, Torkinib (up to 2 μM) did not induce significant cell death (viability > 85% by trypan blue exclusion), indicating low toxicity to normal stem cells [2] |
| References |
[1]. Nat Chem Biol. 2008 Nov;4(11):691-9. [2]. PLoS Biol. 2009 Feb 10;7(2):e38. [3]. Nat Med. 2010 Feb;16(2):205-13. [4]. Blood. 2010 Nov 25;116(22):4560-8. |
| Additional Infomation |
Torkinib is a member of the class of pyrazolopyrimidines that is 1H-pyrazolo[3,4-d]pyrimidine substituted by isopropyl, 5-hydroxyindol-2-yl and amino groups at positions 1, 3 and 4 respectively. It is a potent inhibitor of mTOR and exhibits anti-cancer properties. It has a role as a mTOR inhibitor and an antineoplastic agent. It is a pyrazolopyrimidine, a member of phenols, a member of hydroxyindoles, a biaryl, an aromatic amine and a primary amino compound. Torkinib (PP242) is a first-in-class ATP-competitive mTOR inhibitor that simultaneously targets mTORC1 and mTORC2, distinguishing it from allosteric mTOR inhibitors (e.g., everolimus) which only inhibit mTORC1 [1] - Torkinib overcomes feedback activation of Akt, a common limitation of mTORC1-only inhibitors, by directly inhibiting mTORC2 (the kinase responsible for Akt Ser473 phosphorylation) [3] - In embryonic stem cell research, Torkinib is used as a tool compound to study mTOR-mediated self-renewal, as it suppresses pluripotency without inducing differentiation (unlike PI3K inhibitors) [2] - Torkinib shows efficacy in both solid tumors (colorectal, breast) and hematological malignancies (AML), with particular activity in PTEN-deficient or Akt-hyperactivated subtypes [1][4] - Preclinical studies suggest Torkinib may enhance the efficacy of chemotherapeutic agents (e.g., doxorubicin in AML) by blocking mTOR-mediated survival signaling, though combination data are not reported in the specified literatures [4] |
Solubility Data
| Solubility (In Vitro) |
DMSO: ~62 mg/mL (~201.1 mM) Water: <1 mg/mL Ethanol: ~18 mg/mL (~58.4 mM) |
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (8.11 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 25.0 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: ≥ 2.5 mg/mL (8.11 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 25.0 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. Solubility in Formulation 3: ≥ 2.5 mg/mL (8.11 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 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. Solubility in Formulation 4: 2% DMSO+30% PEG 300+5% Tween 80+ddH2O: 5mg/mL (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.2432 mL | 16.2159 mL | 32.4317 mL | |
| 5 mM | 0.6486 mL | 3.2432 mL | 6.4863 mL | |
| 10 mM | 0.3243 mL | 1.6216 mL | 3.2432 mL |