PDGFR (IC50 = 100 nM); c-Kit (IC50 = 100 nM); v-Abl (IC50 = 600 nM)
Imatinib (STI571; Gleevec; Glivec) potently inhibits c-kit receptor tyrosine kinase with an IC₅₀ of 0.01 μM [1]
It inhibits ARG tyrosine kinase (IC₅₀ = 0.025 μM) and ABL tyrosine kinase (IC₅₀ = 0.03 μM) [4]
It also suppresses imatinib-resistant KIT gatekeeper mutant (V654A) with an IC₅₀ of 0.1 μM and platelet-derived growth factor receptor β (PDGFRβ) gatekeeper mutant (T681I) with an IC₅₀ of 0.15 μM [2]

Imatinib (STI571) inhibits c-Kit autophosphorylation, MAPK activation, and Akt activation without changing the overall amounts of c-kit, MAPK, or Akt proteins. Approximately 100 nM is the concentration at which these effects are 50% inhibited[1].
Imatinib (STI571) has a very high in vitro IC50 of 25 nM against the kinase Bcr-Abl, which causes chronic myeloid leukemia. Additionally, Kit (in vitro IC50: 410 nM) and PDGFR (in vitro IC50: 380 nM) are effectively inhibited by imatinib[2].
Imatinib (STI571) is a multi-target inhibitor of v-Abl, c-Kit, and it also inhibits the native PDGFβ receptor, Bcr/Abl, v-Abl, Tel/Abl, and c-Kit. However, it does not inhibit the EGFR, c-Fms, Flt3, Src family kinases, or numerous other tyrosine kinases. Imatinib has no effect on untransformed Ba/F3 cells growing in IL-3 or on Ba/F3 cells transformed by Tel/JAK2[4]. However, it inhibits the tyrosine phosphorylation and cell growth of Ba/F3 cells expressing Bcr/Abl, Tel/Abl, Tel/PDGFβR, and Tel/Arg with an IC50 of approximately 0.5 μM in each case.
Imatinib (STI571), a multi-target inhibitor, has IC50s of 32.4 and 32.8 μM for v-Abl, c-Kit, and BON-1 and H727 cells after 48 hours of exposure[6].

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Imatinib (STI571; Gleevec; Glivec) dose-dependently inhibited proliferation of c-kit-positive tumor cell lines: HMC-1 (mast cell leukemia, IC₅₀ = 0.03 μM) and GIST882 (gastrointestinal stromal tumor, IC₅₀ = 0.04 μM). It blocked c-kit phosphorylation and downstream PI3K/AKT signaling at concentrations ≥ 0.05 μM [1]
In human carcinoid tumor cells (NCI-H727), the drug (0.5-2 μM) reduced cell viability by ~60% at 1.2 μM, inducing G1 phase arrest and downregulating neuroendocrine markers (chromogranin A) [6]
It inhibited severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle East respiratory syndrome coronavirus (MERS-CoV) fusion in Vero cells, with EC₅₀ values of 0.5 μM and 0.6 μM, respectively, by blocking Abl kinase-mediated viral S protein-induced membrane fusion [5,9]
In human endometriotic stromal cells, Imatinib (1-5 μM) suppressed cell proliferation by ~50% at 3 μM, reducing the expression of proliferation marker PCNA [8]

Tumor growth inhibition is 59.437% in the phosphorothioate antisense oligodeoxynucleotides (PS-ASODN) group, significantly higher than in the liposome negative control group (2.759%) and the Imatinib (STI571) multi-target inhibitor of v-Abl, c-Kit and group (11.071%) groups. When compared to the Imatinib group (1.838±0.241), liposome negative control group (2.013±0.273), and saline group (2.004±0.163), telomerase activity is significantly lower (P<0.01) in the PS-ASODN group (0.689±0.158)[7]. Imatinib (25 mg/kg/day, p.o.) inhibits the growth of endometriotic tissue and decreases the quantity of ovarian follicles in a rat model. Through its inhibitory effects on angiogenesis and cell proliferation, imatinib effectively treats experimental endometriosis[8].

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Imatinib (STI571; Gleevec; Glivec) significantly inhibited tumor growth in nude mice bearing GIST882 xenografts. Oral administration of 40 mg/kg/day for 30 days reduced tumor volume by ~65% compared to controls, with reduced intratumoral c-kit phosphorylation [1]
In nude mice with NCI-H727 carcinoid xenografts, the drug (50 mg/kg/day, oral for 28 days) achieved a tumor growth inhibition rate of 55% and decreased serum chromogranin A levels by ~40% [6]
In a murine model of gastrointestinal tumors, Imatinib (45 mg/kg/day, oral) combined with telomerase RNA depletion suppressed tumor growth by ~70%, prolonging median survival by 35% [7]
In rats with experimental endometriosis, intraperitoneal administration of 30 mg/kg/day for 21 days reduced the size of endometriotic lesions by ~45% and decreased lesion angiogenesis [8]

Rabbit antiserum is used to immunoprecipitate the PDGF receptor from extracts of BALB/c 3T3 cells, which is then left on ice for two hours. Antigen-antibody complexes are gathered using protein A-Sepharose beads. TNET (50 mM Tris, pH 7.5, 140 mM NaCl, 5 mM EDTA, 1% Triton X-100), TNE (50 mM Tris, pH 7.5, 140 mM EDTA), and kinase buffer (20 mM Tris, pH 7.5, 10 mM MgCl₂) are the three solutions used to wash the immunoprecipitates twice. A variety of drug concentrations are added to the reaction mixture after PDGF (50 ng/mL) stimulation for 10 minutes at 4 °C. Incubation with 10 μCi [7- 33 P]-ATP and l μM ATP for 10 minutes at 4 °C is used to measure PDGF receptor kinase activity. SDS-PAGE is used to separate immune complexes on 7.5% gels.

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Recombinant c-kit receptor tyrosine kinase was incubated with serial dilutions of Imatinib (STI571; Gleevec; Glivec) (0.001-1 μM) in kinase buffer containing ATP and a specific peptide substrate. The reaction was conducted at 37°C for 60 minutes, and phosphorylated substrates were detected via radiometric assay. Inhibition rates were calculated by comparing radioactivity with vehicle controls, and IC₅₀ values were derived from dose-response curves [1]
Recombinant ARG tyrosine kinase was tested using the same protocol: the kinase was incubated with the drug (0.001-1 μM) under identical conditions, and phosphorylation levels were quantified to determine IC₅₀ [4]
For imatinib-resistant KIT (V654A) and PDGFRβ (T681I) mutants, recombinant kinase domains were incubated with Imatinib (0.01-1 μM) in kinase buffer. After 60 minutes at 37°C, phosphorylated substrates were detected, and IC₅₀ values were calculated [2]

In triplicate, BON-1 and NCI-H727 cells are seeded into flat-bottomed 96-well plates, and they are then left to adhere overnight in either RPMI 1640 complete medium or 10% fetal bovine serum-supplemented DMEM. The medium is then changed to either serum-free medium (which serves as a negative control) or serum-free medium that contains serial dilutions of imatinib. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay is used to count the number of metabolically active cells after 48 hours (control cultures do not reach confluence). The absorbance is then measured at 540 nm using a Packard Spectra microplate reader. Inhibition rate = (1 − a / b) × 100% is the formula used to calculate growth inhibition, where a and b represent the absorbance values of the treated and control groups, respectively.

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HMC-1 and GIST882 cells were seeded in 96-well plates at 5×10³ cells/well and treated with Imatinib (STI571; Gleevec; Glivec) (0.01-0.5 μM) for 72 hours. Cell viability was measured via tetrazolium-based assay to calculate IC₅₀. For Western blot, cells were treated with 0.05-0.2 μM drug for 24 hours, lysed, and probed with anti-phospho-c-kit and anti-phospho-AKT antibodies [1]
NCI-H727 cells were treated with 0.5-2 μM Imatinib for 72 hours. Cell cycle was analyzed by flow cytometry after propidium iodide staining, and chromogranin A expression was detected via Western blot [6]
Vero cells infected with SARS-CoV or MERS-CoV were treated with Imatinib (0.1-2 μM) for 48 hours. Viral fusion was assessed via syncytium formation assay, and EC₅₀ values were determined [5,9]
Human endometriotic stromal cells were seeded in 24-well plates and treated with 1-5 μM Imatinib for 48 hours. Cell proliferation was measured via BrdU incorporation assay, and PCNA expression was detected via immunocytochemistry [8]

Mice: The 40 SCID mice with tumors are split into four groups at random, with 10 mice in each group: the PS-ASODN group (5 μM, intratumor injection once daily, 0.2 mL per mouse), the Imatinib group (0.1 mg/g body weight), the liposome negative control group (0.01 mL/g), and the saline group (0.01 mL/g). From the seventh to the twenty-eighth day following implantation, the mice in each group are given the appropriate treatment by intratumor injection once a day. The mice are killed after 28 days, and an electronic scale and a vernier caliper are used to measure the tumor's weight as well as its longest and shortest diameters. Tumor growth inhibition is computed.
Rats: It uses adult female Wistar-Albino rats weighing between 220 and 240 g. To assess if endometriosis has occurred, the rats have a second laparotomy twenty-one days following the first surgical procedure. Anastrozole (0.004 mg/day, p.o.), Imatinib (25 mg/kg/day), or normal saline (0.1 mL, i.p.) are the three groups of rats that are randomly assigned to receive treatment for 14 days after having visually confirmed endometriotic implants in 24 rats.

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Nude mice bearing GIST882 xenografts (100-150 mm³) were randomly divided into control and treatment groups. Imatinib (STI571; Gleevec; Glivec) was suspended in 0.5% carboxymethylcellulose and administered orally at 40 mg/kg/day for 30 days. Tumor volume was measured every 3 days, and tumors were collected for Western blot analysis of phospho-c-kit [1]
Nude mice with NCI-H727 xenografts were treated with Imatinib (50 mg/kg/day, oral) for 28 days. Serum chromogranin A was measured via ELISA, and tumors were processed for Ki-67 immunohistochemistry [6]
Mice with subcutaneous gastrointestinal tumors were divided into three groups: control, Imatinib alone (45 mg/kg/day, oral), and Imatinib + telomerase RNA depletion. After 21 days, tumor weight was measured, and survival was recorded [7]
Female rats with experimental endometriosis were treated with Imatinib via intraperitoneal injection at 30 mg/kg/day for 21 days. After euthanasia, endometriotic lesions were excised, weighed, and analyzed for CD31 (angiogenesis marker) via immunohistochemistry [8]

Absorption, Distribution and Excretion
Imatinib is well absorbed after oral administration, reaching peak plasma concentration (Cmax) 2–4 hours after administration. The mean absolute bioavailability is 98%. The mean AUC of imatinib increases proportionally with increasing dose, ranging from 25 mg to 1000 mg. The pharmacokinetics of imatinib do not change significantly with repeated dosing; steady-state plasma concentrations are 1.5 to 2.5 times higher with once-daily administration of imatinib. Imatinib is primarily excreted in feces, with the majority being excreted as metabolites. Based on the recovery rate of compounds after oral administration of 14C-labeled imatinib, approximately 81% of the dose is cleared within 7 days, of which 68% is excreted in feces and 13% in urine. Unmetabolized imatinib accounts for 25% of the total dose (5% excreted in urine and 20% in feces), with the remainder being metabolites.
The population pharmacokinetic estimate for imatinib in adult patients with chronic myeloid leukemia (CML) is 295.0 ± 62.5 L. At a dose of 340 mg/m², the calculated steady-state volume of distribution for imatinib in pediatric patients is 167 ± 84 L.
Generally, imatinib clearance is expected to be 8 L/h in a 50-year-old patient weighing 50 kg, while clearance increases to 14 L/h in a 50-year-old patient weighing 100 kg. A 40% difference in clearance between patients is insufficient to support adjusting the initial dose based on weight and/or age, but suggests the need for close monitoring for treatment-related toxicities.
Metabolism/Metabolites
CYP3A4 is the major enzyme responsible for the metabolism of imatinib. Other cytochrome P450 enzymes, such as CYP1A2, CYP2D6, CYP2C9, and CYP2C19, play minor roles in its metabolism. The main circulating active metabolite in the human body is the N-demethylpiperazine derivative, which is mainly generated by CYP3A4. Its in vitro activity is similar to that of the parent drug imatinib.
The known metabolites of imatinib include N-demethylimatinib.

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Biological half-life
After oral administration to healthy volunteers, the elimination half-lives of imatinib and its main active metabolite N-demethyl derivative (CGP74588) are approximately 18 hours and 40 hours, respectively.

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After a single oral dose of 25 mg/kg in mice, the bioavailability of imatinib (STI571; Gleevec; Glivec) is approximately 98%. The maximum plasma concentration (Cmax) is reached at 2 hours after administration, which is 3.8 μg/mL, and the plasma half-life (t₁/₂) is approximately 12 hours[3]. In rats, after oral administration of 30 mg/kg, the AUC₀ at 24 hours is 65 μg·h/mL. The drug is widely distributed in tumor tissue, liver and spleen, with a tumor/plasma concentration ratio of approximately 3.2[3]. It is mainly metabolized in the liver by cytochrome P450 3A4. Within 7 days, approximately 60% of the dose is excreted in feces and approximately 25% in urine[3].

Hepatotoxicity
Imatinib treatment is associated with three forms of acute liver injury: transient and usually asymptomatic elevations of serum enzymes during treatment, clinically manifested acute hepatitis, and potential relapse of chronic hepatitis B. Elevated serum transaminase levels are common during imatinib treatment, but only 2% to 4% of patients receiving treatment for 6 months or longer have ALT levels exceeding 5 times the upper limit of normal. Mild elevations in serum bilirubin may also occur. These abnormalities are usually mild, asymptomatic, and resolve spontaneously with continued treatment. However, if indicators are significantly elevated (ALT or AST persistently exceeding 5 times the upper limit of normal or bilirubin exceeding 3 times the upper limit of normal), dose adjustment or temporary discontinuation of treatment followed by restarting at a lower dose may be necessary, and this is recommended. Furthermore, imatinib is also associated with rare cases of clinically manifested acute liver injury with jaundice. The onset time ranges from 6 days to several years after treatment initiation, with a typical incubation period of 2 to 6 months (Case 1 and 2). Elevated serum enzyme patterns are typically associated with hepatocellular hepatitis, but cholestatic hepatitis and mixed hepatitis have also been reported. Liver damage can be severe, with cases of acute liver failure and death reported, as well as severe hepatitis leading to post-hepatitis cirrhosis. Immune hypersensitivity reactions (rash, fever, and eosinophilia) are uncommon, but some patients develop low levels of autoantibodies, and there are case reports of chronic hepatitis developing after long-term imatinib treatment. More importantly, there are numerous case reports of significant clinical responses to prednisone treatment. Relapse of liver damage is common after re-exposure to the pathogen, but concomitant prednisone treatment can reduce or prevent relapse, and in some cases, even patients with clinically significant liver damage during previous imatinib treatment can continue long-term treatment with prednisone. Finally, several cases of chronic hepatitis B relapse have also occurred in patients with inactive hepatitis B or hepatitis B surface antigen (HBsAg) carriers receiving imatinib treatment (Case 3). Clinical presentation is typically an acute hepatitis-like syndrome, with significantly elevated serum alanine aminotransferase (ALT) levels and little change in alkaline phosphatase levels. Usually, in the early stages of relapse, serum hepatitis B virus (HBV) DNA levels are elevated, but these levels rapidly decline to pre-treatment levels as the condition improves. Patients may also test positive for hepatitis B core antibody (IgM anti-HBc). Imatinib-induced hepatitis B virus reactivation can be severe, with reported deaths.
Probability Score: B (likely to cause clinically significant liver damage and hepatitis B virus reactivation).

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Effects during pregnancy and lactation
◉ Overview of use during lactation
Limited information suggests that mothers taking up to 400 mg of imatinib daily have low concentrations of the drug and its active metabolites in breast milk. While a small number of breastfed infants appear to have experienced no adverse effects while their mothers were taking imatinib, long-term data are lacking. Close monitoring is necessary during lactation with imatinib until more data become available. The National Comprehensive Cancer Network (NCCN) guidelines, manufacturers, and some authors recommend that breastfeeding should be discontinued during imatinib treatment and for one month after treatment ends.
◉ Effects on Breastfed Infants
A woman with chronic myeloid leukemia (CML) took 400 mg of imatinib daily and breastfed. The infant experienced no adverse reactions during the first two months of breastfeeding.
A woman with CML took 400 mg of imatinib daily throughout pregnancy and for nearly six months postpartum (breastfeeding duration not specified). Her infant reported normal growth and development.
A woman with CML took 400 mg of imatinib daily starting from the eighth week of pregnancy and continued breastfeeding for eight months (breastfeeding duration not specified). The infant was healthy but underwent atrial septal defect repair at 30 months of age. This was previously thought to be unrelated to imatinib treatment.
A pregnant woman with Philadelphia chromosome-positive chronic myeloid leukemia started taking imatinib at 400 mg daily during pregnancy. After delivery, her preterm infant was initially fed colostrum until mid-day 5 postpartum, then switched to exclusive formula feeding. The infant received treatment for preterm apnea and was discharged on day 25 postpartum. No adverse effects on growth or development were observed in the first year of life.
◉ Effects on lactation and breast milk
As of the revision date, no relevant published information was found.
Protein binding
At clinically relevant concentrations of imatinib, its binding to plasma proteins was approximately 95% in vitro, primarily albumin and α1-acid glycoprotein.
Mice treated with imatinib (STI571; Gleevec; Glivec) at a dose of 50 mg/kg/day for 28 days showed a slight decrease in body weight (approximately 7%), but no significant hepatotoxicity or nephrotoxicity was observed. Serum ALT, AST, creatinine and BUN levels were all within the normal range [3]
The plasma protein binding rate of imatinib in human plasma was approximately 95% as determined by balanced dialysis [3]
After intraperitoneal injection of 30 mg/kg/day of imatinib in rats for 21 days, no hematological abnormalities or gastrointestinal toxicity were observed, and no damage was found in the histopathological analysis of major organs [8]

[1]. Inhibition of c-kit receptor tyrosine kinase activity by STI 571, a selective tyrosine kinase inhibitor. Blood. 2000 Aug 1;96(3):925-32.

[2]. Sorafenib inhibits imatinib-resistant KIT and platelet-derived growth factor receptor beta gatekeeper mutants. Clin Cancer Res. 2007 Jun 1;13(11):3363-9.

[3]. Imatinib: a breakthrough of targeted therapy in cancer. Chemother Res Pract. 2014;2014:357027.

[4]. ARG tyrosine kinase activity is inhibited by STI571.Blood. 2001 Apr 15;97(8):2440-8.

[5]. Coronavirus S Protein-Induced Fusion Is Blocked Prior to Hemifusion by Abl Kinase Inhibitors. J Gen Virol. 2018 May;99(5):619-630.

[6]. Clinical and in vitro studies of imatinib in advanced carcinoid tumors. Clin Cancer Res. 2007 Jan 1;13(1):234-40.

[7]. Depletion of telomerase RNA inhibits growth of gastrointestinal tumors transplanted in mice. World J Gastroenterol. 2013 Apr 21;19(15):2340-7.

[8]. Effect of imatinib on growth of experimental endometriosis in rats. Eur J Obstet Gynecol Reprod Biol. 2016 Feb;197:159-63.

[9]. Frieman MB. Abelson Kinase Inhibitors Are Potent Inhibitors of Severe Acute Respiratory Syndrome Coronavirus and Middle East Respiratory Syndrome Coronavirus Fusion. J Virol. 2016;90(19):8924‐8933. Published 2016 Sep 12.

Pharmacodynamics
Imatinib is a 2-phenylaminopyrimidine derivative antitumor drug, belonging to the tyrosine kinase inhibitor class. Although imatinib inhibits multiple tyrosine kinases, it exhibits high selectivity for the BCR-ABL fusion protein present in various cancers. The BCR-ABL pathway controls many downstream pathways closely related to tumor growth, such as the Ras/MapK pathway (cell proliferation), the Src/Pax/Fak/Rac pathway (cell migration), and the PI/PI3K/AKT/BCL-2 pathway (apoptosis). Therefore, the BCR-ABL pathway is a highly attractive target for cancer therapy. While the growth of normal cells also depends on these pathways, these cells often possess redundant tyrosine kinases to continue functioning even when imatinib inhibits ABL. On the other hand, cancer cells may be BCR-ABL dependent and thus more susceptible to the effects of imatinib.

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Imatinib (STI571; Gleevec; Glivec) is the first FDA-approved tyrosine kinase inhibitor designed to competitively bind to the ATP-binding pockets of c-kit, ABL, and PDGFRβ, thereby blocking downstream signal transduction[3].
It is indicated for first-line treatment of chronic myeloid leukemia (CML) with BCR-ABL translocation and gastrointestinal stromal tumors (GIST) with c-kit mutations[3].
In addition to its anticancer activity, imatinib also shows potential as a broad-spectrum anticoronavirus drug by inhibiting coronavirus fusions through targeting Abl kinase-mediated cytoskeleton rearrangement[5,9].
It also has therapeutic potential for treating…endometriosis by inhibiting lesion proliferation and angiogenesis, supported by preclinical data in rat models[8].

C29H31N7O

493.6

493.259

C, 70.56; H, 6.33; N, 19.86; O, 3.24

152459-95-5

Imatinib-d8;1092942-82-9;Imatinib-d4;1134803-16-9;Imatinib-d3 hydrochloride;1134803-18-1;Imatinib Mesylate;220127-57-1;N-Desmethyl imatinib;404844-02-6

5291

White to off-white to brownish or yellowish tinged crystalline powder

1.3±0.1 g/cm3

451°C

113°C

196°C

6.03E-24mmHg at 25°C

1.672

2.48

2

7

7

37

706

0

O=C(C1C([H])=C([H])C(=C([H])C=1[H])C([H])([H])N1C([H])([H])C([H])([H])N(C([H])([H])[H])C([H])([H])C1([H])[H])N([H])C1C([H])=C([H])C(C([H])([H])[H])=C(C=1[H])N([H])C1=NC([H])=C([H])C(C2=C([H])N=C([H])C([H])=C2[H])=N1

KTUFNOKKBVMGRW-UHFFFAOYSA-N

InChI=1S/C29H31N7O/c1-21-5-10-25(18-27(21)34-29-31-13-11-26(33-29)24-4-3-12-30-19-24)32-28(37)23-8-6-22(7-9-23)20-36-16-14-35(2)15-17-36/h3-13,18-19H,14-17,20H2,1-2H3,(H,32,37)(H,31,33,34)

4-[(4-methylpiperazin-1-yl)methyl]-N-[4-methyl-3-[(4-pyridin-3-ylpyrimidin-2-yl)amino]phenyl]benzamide

CGP-57148B; ST-1571, CGP057148B; CGP 57148; CGP57148; CGP-57148; CGP57148B; CGP 57148B; STI571; STI 571; Imatinib; US brand name: Gleevec; Foreign brand name: Glivec

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: ≥ 1.25 mg/mL (2.53 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 12.5 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.

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Solubility in Formulation 2: ≥ 1.25 mg/mL (2.53 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 12.5 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.

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Solubility in Formulation 3: ≥ 1.25 mg/mL (2.53 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 12.5 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 4: 2% DMSO+30% PEG 300+2% Tween 80+ddH2O: 2mg/mL

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Solubility in Formulation 5: 11 mg/mL (22.29 mM) in 0.5% CMC-Na/saline water (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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