Natural phytoalexin; Autophagy; Adenylyl cyclase 0.8 nM (IC50); Mitophagy; IKKβ 1 μM (IC50); DNA polymerase α 3.3 μM (IC50); DNA polymerase δ 5 μM (IC50); Sirtuin

Resveratrol (trans-Resveratrol; SRT501) is one of numerous polyphenolic chemicals found in a range of plant sources. In the vast majority of cases, Resveratrol demonstrates inhibitory/activating effects in the micromolar range, which may be pharmacologically feasible, although targets in the nanomolar range have also been described [1]. MCF-7 cells were plated in DME-F12 media supplemented with 5% FBS and increasing doses of Resveratrol. Control cells were treated with the same volume of vehicle (0.1% ethanol) alone. Resveratrol suppresses the development of MCF-7 cells in a dose-dependent way. Addition of 10 μM Resveratrol resulted in 82% suppression of MCF-7 cell growth after 6 days, but at 1 μM, only 10% inhibition was seen. Cells treated with 10 μM Resveratrol had a doubling time of 60 hours, whereas control cells doubled every 30 hours. Trypan blue exclusion assay demonstrated that at concentrations of 10 μM or below, Resveratrol did not impair cell viability (90% of viable cells), however at a dose of 100 μM, only 50% of cells survived after 6 days of Resveratrol administration. In addition, MCF-7 cells did not undergo apoptosis after incubation with Resveratrol at a dosage of 10 μM, as assessed by the ApoAlert Annexin V Apoptosis Kit [2]. Resveratrol promotes nitric oxide (NO) production in endothelial cells by upregulating the expression of endothelial nitric oxide synthase (eNOS), boosting eNOS enzyme activity, and blocking eNOS uncoupling [7].

In vivo, resveratrol has been shown to increase plasma antioxidant capacity and decrease lipid peroxidation; however, it is difficult to assess whether these effects are direct, or the result of upregulating endogenous antioxidant enzymes.[1]
Although most in vivo studies strongly support a chemopreventive effect of resveratrol, there are notable exceptions in which no benefit has been observed. For example, administration of 1–5 mg per kg (body weight) daily of resveratrol failed to affect the growth or metastasis of breast cancer in mice, despite promising in vitro results17. Dosage, delivery method, tumour origin and other components of the diet could all contribute to the efficacy of resveratrol treatment. Overall, in vivo studies clearly show great promise for this molecule in the treatment of cancers. Several Phase I clinical trials are currently underway for oral resveratrol in humans at doses as high as 7.5 g per day, including National Cancer Institute-sponsored studies at the University of Michigan, USA, and the University of Leicester.[1]
In vivo, resveratrol has been shown to increase expression of both endothelial and inducible nitric oxide synthase (eNOS and iNOS, respectively).[1]
Because resveratrol is an effective inhibitor of cyclooxygenase activity in vivo20,22,28, its anti-inflammatory properties have been investigated. [1]
Treatment with resveratrol (trans-Resveratrol; SRT501) at 50 mg/kg (195.5±124.8 mm3; P<0.05) or 100 mg/kg reduced the mean tumor volume (81.7±70.5 mm3; P<0.001). Tumor mass and volume have a strong correlation.

Insulin-like Growth Factor 1 Enzyme-Linked Immunosorbent Assay[2]
The concentration of IGF-1 in the cell supernatant was measured using Human IGF-I/IGF-1 DuoSet ELISA. For the measurement, microglia (HMC3) were seeded in 6-well plates (80,000 cells/1 mL medium/well) and treated with resveratrol (100 µM) for 24 h. Afterwards, cell supernatants were collected and processed according to manufacturer instructions.

---

Glucose Uptake Quantification[2]
For analysis of glucose uptake, microglia (HMC3) were seeded in white 96-well plates (5000 cells/100 µL medium/well). Upon 24 h of resveratrol (100 µM) treatment, glucose uptake from the medium was measured in technical duplicates using the non-radioactive Glucose Uptake-Glo™ Assay according to the manufacturer’s instructions. Plates were read using the TECAN GENios microplate reader.

---

TMRE-Mitochondrial Membrane Potential Assay[2]
Mitochondrial activity was investigated using the TMRE-Mitochondrial Membrane Potential Assay Kit according to manufacturer’s instructions. Microglia (HMC3) were seeded in 24-well plates (7500 cells/250 µL medium/well). Imaging was carried out upon the 24 h resveratrol (100 µM) treatment using the Keyence BZx800 Fluorescence Microscope. The fluorescence intensity of two areas from each experiment was quantified using ImageJ

Inflammasome Activity Assays[2]
For the quantification of inflammasome activity, microglia (HMC3) were seeded in white 96-well plates (8000 cells/100 µL medium/well). Upon 6 h of resveratrol (100 µM) treatment, inflammasome activity was measured in technical duplicates using the Caspase-Glo® 1 Inflammasome Assay according to the manufacturer’s instructions. Plates were read using the TECAN GENios microplate reader.

---

Scanning Electron Microscopy (SEM)[2]
Microglia (HMC3) were seeded on 24 mm × 12 mm coverslips that were placed in 6-well plates (60,000 cells/1 mL medium/well). After 24 h resveratrol (100 µM) treatment cells were washed with PBS and fixed for 30 min in 3% glutaraldehyde. In the next step, samples were washed with PBS and kept in a 2% osmium solution for 20 min. Subsequently, all water was removed by placing samples in increasing ethanol concentrations (30–100%), and critical point drying was carried out using a CPD 030. Finally, samples were coated with an SCD 050 sputter coater for 50 s and imaged using a JSM-IT200.

---

Proliferation[2]
Proliferation was determined by counting the microglia (HMC3) seeded in 6-well plates (80,000 cells/1 mL medium/well) using the T20 Automated Cell Counter after 24 h of resveratrol (100 µM) treatment. Proliferation was calculated as an n-fold amount of the initially seeded cell number.

The purpose of this study was to investigate the protective effect of low-dose trans-resveratrol (trans-RSV) on diabetes-induced retinal ganglion cell (RGC) degeneration and its possible mechanism. Methods: A streptozotocin-induced diabetic mouse model was established and treated with or without trans-RSV intragastric administration (10 mg/kg body weight/day) for 12 weeks. Oscillatory potentials (Ops) of the dark-adapted electroretinogram (ERG) were recorded. The number of RGCs was detected by Tuj1 and TUNEL staining. The apoptosis markers in the retina were analyzed by Western blot. The cross sections of optic nerves were observed by transmission electron microscopy. In addition, mouse neuroblastoma N2a cells were injured by high-glucose (HG) treatment. Cell viability and apoptosis were measured with or without low-dose trans-RSV treatment. The intracellular localization of tyrosyl transfer-RNA synthetase (TyrRS) was observed in both mouse retinas and N2a cells. The effects of low-dose trans-RSV on the binding of TyrRS to the transcription factor c-Jun and the binding of c-Jun to pro-apoptotic genes were analyzed by co-IP and ChIP assays in HEK 293 cells. Results: Trans-RSV relieved electrophysiological injury of retinas and inhibited RGC apoptosis in diabetic mice. It also protected N2a cells from HG-induced apoptosis. Additionally, it promoted TyrRS nuclear translocation in both diabetic mouse retinas and HG-treated N2a cells. Trans-RSV promoted TyrRS binding to c-Jun, inhibited the phosphorylation of Ser-63 of c-Jun, and downregulated pro-apoptotic gene transcription. Conclusions: Low-dose trans-RSV can ameliorate diabetes-induced RGC degeneration via the TyrRS/c-Jun pathway. It can promote TyrRS nuclear translocation and bind to c-Jun, downregulating c-Jun phosphorylation and downstream pro-apoptotic genes.https://pubmed.ncbi.nlm.nih.gov/37261387/

---

Dissolved insodium lactate buffer (50 mM, pH 4.0); 100 mg/kg; i.p. injection

Human ovarian xenografts PA-1

Absorption, Distribution and Excretion
High absorption but very low bioavailability.
... Glycosylated resveratrol is more stable and more soluble and readily absorbed in the human gastrointestinal tract ... In humans, following its absorption, it is readily metabolized in the liver ... to water-soluble trans-resveratrol-3-O-glucuronide and trans-resveratrol-3-O-sulfate, accounting for its predominant urine excretion ... Compared to other known polyphenols, such as quercetin and catechin, trans-resveratrol is well absorbed much more efficiently following oral administration to humans ...
... Resveratrol is absorbed from the gastrointestinal tract following its ingestion ...
... Preclinical studies in rats, using HPLC methods, have suggested that intragastric administration of 20 mg/kg trans-resveratrol generated peak values of 1.2 uM in plasma ... In a separate study, male rats treated with 300, 1000, and 3000 mg/kg body weight per day were reported to achieve plasma concentrations of 576, 991, and 2728 ng/mL, respectively, and whereas in females, it was 333, 704, and 1137 ng/mL ... A plasma concentration of approximately 1.1 ug/mL was determined to be approximately 5 uM ... A single oral administration of (14)C-trans-resveratrol to male Balb/c mice showed preferential binding of radio-labeled resveratrol in the stomach, liver, kidney, intestine, bile, and urine, and penetrated the tissues of the liver and kidney ... Both the parent compound and the Phase-II metabolites were also detected in these tissues ... In humans, 24.6% of the oral dose administered appeared in the urine, including metabolites ... whereas after intragastric administration to rodents, only 1.5% of resveratrol reached the plasma compartment ...
In a human melanoma xenograft model ... The resveratrol content in the skin of these mice, measured 5 min after a bolus of 75 mg/kg introduced, was found to be 21 nmol/g and 4.67 nmol/g in glucuronide-conjugate forms. A measurable amount of resveratrol was found in the tumors, although it was less than the amount found in the skin ...
For more Absorption, Distribution and Excretion (Complete) data for RESVERATROL (12 total), please visit the HSDB record page.

---

Metabolism / Metabolites
Hepatic. Rapidly metabolized and excreted.
... This study ... examined the absorption, bioavailability, and metabolism of (14)C-resveratrol after oral and iv doses in six human volunteers. The absorption of a dietary relevant 25-mg oral dose was at least 70%, with peak plasma levels of resveratrol and metabolites of 491 + or - 90 ng/mL (about 2 uM) and a plasma half-life of 9.2 + or - 0.6 hr. However, only trace amounts of unchanged resveratrol (<5 ng/mL) could be detected in plasma. Most of the oral dose was recovered in urine, and liquid chromatography/mass spectrometry analysis identified three metabolic pathways, ie, sulfate and glucuronic acid conjugation of the phenolic groups and, interestingly, hydrogenation of the aliphatic double bond, the latter likely produced by the intestinal microflora. Extremely rapid sulfate conjugation by the intestine/liver appears to be the rate-limiting step in resveratrol's bioavailability.
In plants, it mostly exists in glycosylated piceid forms (3-O-B-D-glucosides). Other minor conjugated forms containing 1 to 2 methyl groups (pterostilbene), a sulfate group (trans-resveratrol-3-sulfate) or a fatty acid have also been identified. Glycosylation is known to protect resveratrol from oxidative degradation, and glycosylated resveratrol is more stable ... In humans, following its absorption, it is readily metabolized in the liver by Phase-2 drug-metabolizing enzymes to water-soluble trans-resveratrol-3-O-glucuronide and trans-resveratrol-3-O-sulfate, accounting for its predominant urine excretion ...
... Resveratrol was metabolized in humans into two metabolites, which were characterized as resveratrol-3-O- and 4'-O-glucuronides ...
... In two 8-week long feeding experiments with rats, a low-resveratrol diet containing 50 mg resveratrol per kg body weight (bw) and day and a high-resveratrol diet with 300 mg per kg bw and day were administered. ... The level of resveratrol and its metabolites in the feces, urine, plasma, liver, and kidneys was identified and quantitated by high-performance liquid chromatography-diode array detection (HPLC-DAD) using synthesized resveratrol conjugate standards. ... The formation of trans-resveratrol-3-sulfate, trans-resveratrol-4'-sulfate, trans-resveratrol-3,5-disulfate, trans-resveratrol-3,4'-disulfate, trans-resveratrol-3,4',5-trisulfate, trans-resveratrol-3-O-beta-D-glucuronide, and resveratrol aglycone was detected by HPLC analysis, depending on the biological material. ...
Resveratrol has known human metabolites that include Resveratrol 3-O-glucuronide.

---

Biological Half-Life
Pharmacokinetics of trans-resveratrol in its aglycone (RES(AGL)) and glucuronide (RES(GLU)) forms were studied following intravenous (15 mg/kg i.v.) and oral (50 mg/kg p.o.) administration of trans-resveratrol in a solution of beta-cyclodextrin to intact rats ... After i.v. administration, plasma concentrations of RES(AGL) declined with a rapid elimination half-life (T(1/2), 0.13 hr), followed by sudden increases in plasma concentrations 4 to 8 hr after drug administration. These plasma concentrations resulted in a significant prolongation of the terminal elimination half-life of RES(AGL) (T(1/2TER), 1.31 hr). RES(AGL) and RES(GLU) also displayed sudden increases in plasma concentrations 4 to 8 hr after oral administration, with T(1/2TER) of 1.48 and 1.58 hr, respectively ...
Resveratrol ... has a plasma half-life of 8 to 14 min; the metabolites /(trans-resveratrol-3-O-glucuronide and trans-resveratrol-3-O-sulfate)/ have a plasma half-life of about 9.2 hours ...
... A dietary relevant 25-mg oral dose /of (14)C-resveratrol in six human volunteers resulted in/ ... a plasma half-life of 9.2 + or - 0.6 hr ...
A 25-mg oral dose /of (14)C-resveratrol had/ a plasma half-life of 9.2 + or - 0.6 hr. ...
Trans-resveratrol half-life /in human plasma/ was 1-3 hr following single-doses and 2-5 hr following repeated dosing.

Hepatotoxicity
Liver injury attributable to resveratrol has not been reported. In trials of resveratrol in human subjects, there have been no reports of serum enzyme elevations or clinically apparent liver injury. Thus, hepatotoxicity due to resveratrol must be rare, if it occurs at all.
Likelihood score: E (unlikely cause of clinically apparent liver injury).
Drug Class: Herbal and Dietary Supplements

---

Effects During Pregnancy and Lactation
◉ Summary of Use during Lactation
Resveratrol (3,4',5-trans-trihydroxystilbene) is an antioxidant found in numerous plant species and in red wine. Resveratrol has no specific lactation-related uses. In general, it is used to prevent heart disease, cancer, and other diseases associated with aging, although high-quality studies are lacking. Resveratrol appears to be relatively free from adverse reactions. However, no data exist on the excretion of resveratrol into breastmilk or on the safety and efficacy of resveratrol in nursing mothers or infants. Resveratrol supplements usually contain hundreds of times the amounts found in wine or other foods, so their safety during breastfeeding cannot be assured. It is probably best to avoid the use of red wine as a source of resveratrol during breastfeeding. Refer to the LactMed record on Alcohol for details.
Dietary supplements do not require extensive pre-marketing approval from the U.S. Food and Drug Administration. Manufacturers are responsible to ensure the safety, but do not need to prove the safety and effectiveness of dietary supplements before they are marketed. Dietary supplements may contain multiple ingredients, and differences are often found between labeled and actual ingredients or their amounts. A manufacturer may contract with an independent organization to verify the quality of a product or its ingredients, but that does not certify the safety or effectiveness of a product. Because of the above issues, clinical testing results on one product may not be applicable to other products. More detailed information about dietary supplements is available elsewhere on the LactMed Web site.
◉ Effects in Breastfed Infants
Relevant published information was not found as of the revision date.
◉ Effects on Lactation and Breastmilk
Relevant published information was not found as of the revision date.

---

Protein Binding
Strong affinity towards protein binding.

---

Interactions
... /Resveratrol exhibited/ dose-dependent inhibition of the mutagenic response induced by treatment of Salmonella typhimurium strain TM 677 with 7,12-dimethyl-benzanthracene (DMBA) ...
... Resveratrol ... enhanced the anti-tumor effect of 5-FU ...
In neuronal-like cells, such as the human neuroblastoma SH-SY5Y resveratrol was shown to inhibit caspase 7 activation, as well as degradation of poly-(ADP-ribose)-polymerase, which occur in cells exposed to paclitaxel, an anti-cancer drug ... Resveratrol was shown to induce S-phase arrest, preventing SH-SY5Y from entering mitosis, the phase of the cell cycle in which paclitaxel exerts its activity ... Phosphorylation of Bcl-2 and JNK/SAPK, which specifically occurs after paclitaxel exposure, was reversed by resveratrol ...
The combined effects of resveratrol have been tested with Ara-C or tiazofurin, both antimetabolites, and showed synergistic growth inhibition and apoptosis induction in HL-60 cells ...
For more Interactions (Complete) data for RESVERATROL (22 total), please visit the HSDB record page.

[1]. Nat Rev Drug Discov. 2006 Jun;5(6):493-506.
[2]. Antioxidants (Basel). 2023 Jun 9;12(6):1248.

Therapeutic Uses
Anti-Inflammatory Agents, Non-Steroidal; Anticarcinogenic Agents; Antimutagenic Agents; Antineoplastic Agents, Phytogenic; Antioxidants; Enzyme Inhibitors; Platelet Aggregation Inhibitors
/EXPL THER/ The antioxidant potency of three natural polyphenols, resveratrol, curcumin, and genistein, was compared by using the two human models: oxymodified with H2O2 and homocysteine (Hcy) G proteins in the postmortem frontal cortex (FC) membranes of age-matched control and Alzheimer's disease (AD) subjects; and Cu(2+)-induced oxidation of plasma low-density lipoproteins (LDL). In Co, 3-10 uM polyphenols dose-dependently depressed the G protein 25% stimulation induced by 10 uM H2O2 or 500 uM Hcy. Resveratrol revealed significantly higher antioxidativity than curcumin or genistein. In AD, the antioxidativity of polyphenols showed no significant differences. Polyphenols (1 uM) significantly increased the LDL oxidation lag time (oxyresistance) as compared with control, the effect of resveratrol being most potent. Due to the dual antioxidant mechanism, the investigated polyphenols, particularly resveratrol, should have preferences for the preventive-therapeutic use in age-related oxidative stress-based pathologies.
/EXPL THER/ Arthritis, an inflammation of the joints, is usually a chronic disease that results from dysregulation of pro-inflammatory cytokines (e.g. tumor necrosis factor and interleukin-1beta) and pro-inflammatory enzymes that mediate the production of prostaglandins (e.g. cyclooxygenase-2) and leukotrienes (e.g. lipooxygenase), together with the expression of adhesion molecules and matrix metalloproteinases, and hyperproliferation of synovial fibroblasts. All of these factors are regulated by the activation of the transcription factor nuclear factor-kappaB. Thus, agents that suppress the expression of tumor necrosis factor-alpha, interleukin-1beta, cyclooxygenase-2, lipooxygenase, matrix metalloproteinases or adhesion molecules, or suppress the activation of NF-kappaB, all have potential for the treatment of arthritis. Numerous agents derived from plants can suppress these cell signaling intermediates, including curcumin (from turmeric), resveratrol (red grapes, cranberries and peanuts), tea polyphenols, genistein (soy), quercetin (onions), silymarin (artichoke), guggulsterone (guggul), boswellic acid (salai guggul) and withanolides (ashwagandha). Indeed, several preclinical and clinical studies suggest that these agents have potential for arthritis treatment. Although gold compounds are no longer employed for the treatment of arthritis, the large number of inexpensive natural products that can modulate inflammatory responses, but lack side effects, constitute 'goldmines' for the treatment of arthritis.
/EXPL THER/ Resveratrol, a red wine polyphenol, is known to protect against cardiovascular diseases and cancers, as well as to promote antiaging effects in numerous organisms. It also modulates pathomechanisms of debilitating neurological disorders, such as strokes, ischemia, and Huntington's disease. The role of resveratrol in Alzheimer's disease is still unclear, although some recent studies on red wine bioactive compounds suggest that resveratrol modulates multiple mechanisms of Alzheimer's disease pathology ...
For more Therapeutic Uses (Complete) data for RESVERATROL (12 total), please visit the HSDB record page.

---

Drug Warnings
Pregnant women and nursing mothers should avoid the use of resveratrol-containing supplements. They should also avoid the use of wine as a resveratrol source. Purple grape juice is a good and safe source of resveratrol, as well as other polyphenolic antioxidants.
Resveratrol is contraindicated in those hypersensitive to any component of resveratrol-containing product.

---

Pharmacodynamics
Resveratrol, a phytoalexin, has been found to inhibit herpes simplex virus types 1 and 2 (HSV-1 and HSV-2) replication in a dose-dependent, reversible manner, although this is only one of its many pharmaceutical properties. In some countries where there is higher consumption of red wine, there appears to be a lower incidence of heart disease. Other benefits of resveratrol include its anti-inflammatory and antioxidant effects. In preclinical studies, Resveratrol has been found to have potential anticancer properties.

C14H12O3

228.24

228.078

C, 73.67; H, 5.30; O, 21.03

501-36-0

Resveratrol;501-36-0

445154

White to off-white solid powder

1.4±0.1 g/cm3

449.1±14.0 °C at 760 mmHg

253-255°C

222.3±14.7 °C

0.0±1.1 mmHg at 25°C

1.763

3.14

3

3

2

17

246

0

C1=CC(=CC=C1/C=C/C2=CC(=CC(=C2)O)O)O

LUKBXSAWLPMMSZ-OWOJBTEDSA-N

InChI=1S/C14H12O3/c15-12-5-3-10(4-6-12)1-2-11-7-13(16)9-14(17)8-11/h1-9,15-17H/b2-1+

(E)-5-(4-hydroxystyryl)benzene-1,3-diol

SRT-501; RM-1812; SRT 501; RM 1812; SRT501; RM1812; trans-Resveratrol; CA1201; CA-1201; CA 1201; Resvida; Vineatrol 20M.

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: 5 mg/mL (21.91 mM) in 10% EtOH + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 50.0 mg/mL clear EtOH 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: 5 mg/mL (21.91 mM) in 10% EtOH + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 50.0 mg/mL clear EtOH 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: ≥ 5 mg/mL (21.91 mM) (saturation unknown) in 10% EtOH + 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 50.0 mg/mL clear EtOH stock solution to 900 μL of corn oil and mix well.

---

Solubility in Formulation 4: ≥ 2.5 mg/mL (10.95 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 5: ≥ 2.5 mg/mL (10.95 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 6: ≥ 2.5 mg/mL (10.95 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 7: ≥ 2.5 mg/mL (10.95 mM) (saturation unknown) 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 8: ≥ 2.5 mg/mL (10.95 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), clear solution.
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 9: 2% DMSO+30% PEG 300+ddH2O: 5mg/mL

---

Solubility in Formulation 10: 12.5 mg/mL (54.77 mM) in 50% PEG300 50% Saline (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.

---

Solubility in Formulation 11: 16.67 mg/mL (73.04 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.

---

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