Nefazodone causes oxidative stress (shown by glutathione depletion) and disturbs the potential of the mitochondrial membrane, which results in cell death [2]. HepG2 cells cultured on glucose and galactose show 100% ATP depletion in response to nevazodone (200 μM) given for 24 hours [2]. HepG2 oxygen consumption (OCR) is considerably inhibited by nevazodone (6.25, 12.5, and 25 μM; 0-120 min) [2].

Nefazodone (10 mg/kg), administered subcutaneously over a 16-day period, effectively counteracts the deleterious effects of stress on the immune system in mice [3].

Animal/Disease Models: Female balb/c (Bagg ALBino) mouse (7-12 weeks old; stress model; exposed to 100 dB broadband noise daily for 5 seconds per minute for 1 or 3 hrs (hrs (hours)) around midnight) [3]
Doses: 10 mg/kg
Route of Administration: subcutaneous injection; lasts for 16 days.
Experimental Results: Reduces stress-induced decreases in the number of thymus, spleen and peripheral blood cells.

Absorption, Distribution and Excretion
Nefazodone is rapidly and completely absorbed. Its absolute bioavailability is low (approximately 20%). Following oral administration, nefazodone is primarily metabolized via N-dealkylation and aliphatic and aromatic hydroxylation; less than 1% of the administered nefazodone is excreted unchanged in the urine. 0.22 to 0.87 L/kg Breast Milk/ In two subjects, one took 50 mg twice daily, and the other took 50 mg in the morning and 100 mg in the evening. Plasma trough concentrations were both below 50 ng/mL, while the corresponding breast milk concentrations were 687 ng/mL and 213 ng/mL, respectively. Breast Milk/ …In a woman taking 200 mg nefazodone twice daily, the concentrations of nefazodone in breast milk and plasma (trough concentration) were 57 ng/mL and 617 ng/mL, respectively. The breast milk to plasma concentration ratio was 0.09. Nefazodone is excreted into breast milk. This study aimed to investigate whether adverse reactions in preterm infants could be attributed to nefazodone exposure through breast milk. A 35-year-old (60 kg) Caucasian female infant (female, 2.1 kg, 36 weeks gestational age) receiving nefazodone 300 mg/day was admitted to the hospital with lethargy, listlessness, inability to maintain a normal body temperature, and feeding difficulties. Breast milk exposure to nefazodone was considered after ruling out other more likely diagnoses. Concentration-time profiles of nefazodone and its metabolites (triazoledione, hydroxynefazodone, and m-chlorophenylpiperazine) in maternal plasma and breast milk were quantitatively analyzed using high-performance liquid chromatography (HPLC). The results showed that the infant ingested only 0.45% of the mother's weight-adjusted daily nefazodone dose and its active metabolites (in nefazodone equivalents) through breast milk. ...
For more complete data on the absorption, distribution, and excretion of nefazodone (7 types), please visit the HSDB record page.

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Metabolism/Metabolites
Hepatitis.
...A 16-year-old female received 2.4 g of nefazodone. The terminal elimination half-life of nefazodone was 8.3 hours, and that of its metabolite hydroxynefazodone was 14.6 hours. Blood pressure-time curves showed an 18-hour period of hypotension. Systolic blood pressure was significantly correlated with hydroxynefazodone (R² = 0.602). Despite the hypotension, heart rate remained between 56 and 66 beats/min over 30 hours. QT interval was significantly correlated with both nefazodone (R² = 0.911) and hydroxynefazodone (R² = 0.797), but QTc interval was not significantly correlated with drug concentration. This study aimed to investigate whether adverse reactions in preterm newborns could be attributed to exposure to nefazodone in breast milk. A 35-year-old (60 kg) Caucasian female infant (female, 2.1 kg, corrected gestational age 36 weeks) taking nefazodone 300 mg/day was admitted to the hospital due to lethargy, listlessness, inability to maintain a normal body temperature, and feeding difficulties. High-performance liquid chromatography (HPLC) was used to quantitatively analyze the concentration-time curves of nefazodone and its metabolites (triazolidinedione, hydroxynefazodone, and m-chlorophenylpiperazine) in the mother's plasma and breast milk. Calculations showed that the infant ingested only 0.45% of the mother's weight-corrected daily nefazodone dose and its active metabolites through breast milk (in nefazodone equivalents). This study used nefazodone, an antidepressant with a well-defined metabolic profile, as an example to explore the application value of multivariate analysis in in vitro metabolite identification studies. The chromatographic conditions were chosen to simulate the conditions used in high-throughput screening of the microsomal stability of new chemical entities. Principal component analysis (PCA) was used to evaluate the molecular ions and retention times of human liver microsomal samples containing/without nefazodone. PCA scores and loading plots revealed the separation of the drug and its corresponding metabolites, as well as the associated target ions. Subsequently, tandem mass spectrometry (MS/MS) analysis of the target ions allowed for spectral analysis and resolution to identify nefazodone and its metabolites. Comparison of PCA-identified nefazodone metabolites with those identified using conventional metabolite identification methods showed excellent correlation when similar analytical methods were employed. In a human liver microsomal incubation system supplemented with β-nicotinamide adenine dinucleotide phosphate (NADPH), 15 nefazodone metabolites were identified, encompassing almost all previously reported major metabolites. Of the 15 metabolites, 8 originated from N-dealkylation and N-dephenylation of the N-substituted 3-chlorophenylpiperazine group in nefazodone, 6 from monohydroxylation and dihydroxylation, and 1 from the Bayer-Villiger oxidation of the ethyltriazolone moiety in nefazodone. Following oral administration of nefazodone, it is extensively metabolized primarily through N-dealkylation and aliphatic and aromatic hydroxylation, with less than 1% excreted unchanged in the urine. This paper characterizes three metabolites identified in plasma: hydroxynefazodone (HO-NEF), m-chlorophenylpiperazine (mCPP), and a triazole dione metabolite. This paper describes a complete analysis and characterization of the in vitro metabolites produced by the antidepressant nefazodone (NEF) in human liver microsomes (HLM). The characterization of these three new metabolites revealed two novel metabolic pathways (biotransformation) of NEF. The three novel metabolites included two (M24 and M25) formed by N-dealkylation between a triazolone and a propyl unit, and one (M26) formed by O-dearylation of a phenoxyethyl unit. These metabolites were initially detected by a 4000 Q-Trap mass spectrometer, followed by precise mass measurements using an LTQ-Orbitrap mass spectrometer to confirm their presence. Both instruments provide complete in vitro metabolite information in a single liquid chromatography/tandem mass spectrometry (LC/MS/MS) analysis, but each has its advantages and disadvantages. A significant drawback of the 4000 Q-Trap is the deterioration of isotopic mode quality in the enhanced mass spectrometry (EMS) spectrum when used as a full scan triggering multiple product-dependent ion scans. This problem is particularly pronounced for trace metabolites with low signal intensity. On the other hand, the LTQ-Orbitrap maintains excellent isotopic modes when used as a full scan. A total of 26 metabolites were detected and identified. Analysis of repeated incubation experiments at different time points also confirmed the formation of these new metabolites. Known metabolites of nefazodone include hydroxynefazodone, 1-(3-chlorophenyl)piperazine, 5-ethyl-4-(2-phenoxyethyl)-2-(3-hydroxypropyl)-2H-1,2,4-triazol-3(4H)-one, and p-hydroxynefazodone. It is primarily metabolized in the liver. Elimination pathway: After oral administration of nefazodone, it is primarily metabolized via N-dealkylation and aliphatic and aromatic hydroxylation; less than 1% of nefazodone is excreted unchanged in the urine. Half-life: 2-4 hours. …A 16-year-old female ingested 2.4 g of nefazodone. ...The terminal elimination half-life of nefazodone is 8.3 hours, and that of its metabolite hydroxynefazodone is 14.6 hours. ...In a four-period crossover study, 16 participants received either clinically relevant doses of venlafaxine, nefazodone, or sertraline for 8 days, or fluoxetine for 11 days. A washout period of 7 to 14 days was provided between treatment regimens, with fluoxetine always being the last antidepressant administered. ...Nefazodone is also the only antidepressant that significantly alters the distribution of alprazolam (ALPZ) in the body, showing a decreased area under the concentration-time curve (AUC) (P < 0.01) and a prolonged elimination half-life (16.4 hours vs. 12.3 hours; P < 0.05) compared to baseline. The half-life of nefazodone is 2 to 4 hours.

Toxicity Summary
Identification and Uses: Nefazodone is a solid antidepressant. Human Studies: Post-marketing experience has shown reports of overdose with nefazodone alone and in combination with alcohol and/or other substances. While there have been a few reports of death due to nefazodone overdose (primarily in combination with alcohol and/or other substances), a causal relationship between these deaths and nefazodone has not been established. In pre-marketing clinical studies, there were 7 reports of overdose with nefazodone alone or in combination with other drugs. None of these patients died. The recommended intake range for nefazodone is 1000 mg to 11200 mg. Common symptoms of nefazodone overdose include nausea, vomiting, and drowsiness. One patient who ingested 2000 to 3000 mg of nefazodone, along with mesoparmore and alcohol, reported experiencing seizures. Nefazodone treatment is associated with liver abnormalities ranging in severity from asymptomatic, reversible elevations in serum transaminases to liver failure (ultimately requiring liver transplantation and/or death). Currently, there is no method to predict which patients are likely to develop liver failure. Patients with active liver disease should generally not receive nefazodone treatment. This drug does not increase the incidence of major malformations during pregnancy in humans by 1% to 3% above baseline. Animal studies: There is no evidence of nefazodone carcinogenicity. Feeding rats and mice with nefazodone via diet for two consecutive years at daily doses up to 200 mg/kg and 800 mg/kg, respectively, did not result in an increased incidence of tumors. Reproductive studies were conducted in pregnant rabbits and rats at daily doses up to 200 and 300 mg/kg, respectively (approximately 6 and 5 times that of pregnant rabbits, respectively). No offspring malformations were observed with nefazodone treatment. However, increased early pup mortality and decreased pup weight were observed in rats. A rat fertility study showed that a dose of 200 mg/kg/day resulted in a slight decrease in fertility. Nefazodone has been confirmed to be non-genotoxic based on the following tests: bacterial mutation assay, DNA repair assay in cultured rat hepatocytes, mammalian mutation assay in Chinese hamster ovary cells, in vivo cytogenetics assay in rat bone marrow cells, and rat dominant lethality assay. In the serotonergic system, nefazodone acts as an antagonist of the type 2 serotonin (5-HT2) receptor. Nefazodone acts on postsynaptic receptors and, like fluoxetine antidepressants, inhibits the reuptake of presynaptic serotonin (5-HT). These mechanisms increase the amount of serotonin that can interact with 5-HT receptors. In the norepinephrine system, nefazodone has a small inhibitory effect on norepinephrine uptake. Nefazodone also antagonizes α1-adrenergic receptors, producing sedative, muscle relaxant, and various cardiovascular effects. Nefazodone has no significant affinity for benzodiazepine, cholinergic, dopaminergic, histaminergic, or β- or α2-adrenergic receptors.

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Interactions
Simvastatin is an hydroxymethylglutaryl-CoA reductase inhibitor commonly used to treat hyperlipidemia. In most patients, it is a safe and effective drug when used appropriately. Serious side effects: Rhabdomyolysis may occur rarely in patients taking simvastatin, especially at higher doses or in combination with drugs that increase simvastatin blood concentrations. We describe a case of rhabdomyolysis in a patient who developed it after titrating simvastatin to 80 mg and simultaneously switched from an antidepressant to nefazodone. We found only two similar cases in the literature, both published as reader letters in two different journals. We report this case to enrich the relevant literature and improve clinicians' awareness of this drug interaction for monitoring. This study aimed to evaluate the potential interaction between the novel antidepressant nefazodone and alprazolam after single and multiple doses in 48 healthy male volunteers through a randomized, double-blind, parallel-group, placebo-controlled trial. Twelve participants received a placebo twice daily, and were given either 1 mg alprazolam, 200 mg nefazodone, or 1 mg alprazolam combined with 200 mg nefazodone twice daily for 7 days. Serial blood samples were collected on days 1 and 7 after administration, and on the mornings of days 4, 5, and 6 before administration. The concentrations of alprazolam and its metabolites α-hydroxyalprazolam (AOH) and 4-hydroxyalprazolam (4OH), as well as nefazodone and its metabolites hydroxynefazodone (HO-nefazodone), m-chlorophenylpiperazine (mCPP), and a triazole dione metabolite (diketone) were determined using a validated high-performance liquid chromatography (HPLC). Plasma concentrations of alprazolam, 4OH, nefazodone, HO-nefazodone, mCPP, and diketone reached steady state on day 4. Non-compartmental pharmacokinetic analysis showed that, at steady state, when alprazolam was co-administered with nefazodone, the Cmax and AUCtau values of alprazolam increased significantly by approximately 2-fold, while the Cmax and AUCtau values of 4-hydroxynefazodone decreased significantly by 40% and 26%, respectively. Co-administration of alprazolam with nefazodone had no effect on the single-dose or steady-state pharmacokinetics of nefazodone, hydroxynefazodone, or diketone. However, compared to nefazodone alone, co-administration of alprazolam with nefazodone significantly increased the mean steady-state Cmax and AUCtau values of mCPP by approximately 3-fold, and the t1/2 value by approximately 2-fold. Competitive inhibition of cytochrome P450 3A4 metabolism by alprazolam and nefazodone may be the reason for this phenomenon. There is a pharmacokinetic interaction between alprazolam and nefazodone when used together. No dose adjustment of nefazodone is necessary when using nefazodone and alprazolam together. However, because nefazodone increases plasma alprazolam concentrations, it is recommended to reduce the alprazolam dose when using the two drugs together. Tacrolimus (FK-506) is an important immunosuppressant, most commonly used to maintain immunosuppression to prevent acute cellular organ rejection. A 57-year-old woman with end-stage renal disease (presumably secondary to chronic glomerulonephritis) underwent a living-related kidney transplant. She tolerated the surgery well and was discharged on postoperative day 5. She maintained stable condition using prednisone, azathioprine, and tacrolimus. Two years after the transplant, she started taking nefazodone 50 mg. The patient was prescribed twice-daily oral nefazodone for depression. One week after starting nefazodone, the patient experienced headache, confusion, and blurred vision (gray areas in the visual field), but ophthalmological examination was unremarkable. The patient's serum creatinine increased to 2.2 mg/dL (baseline value 1.5 mg/dL), and trough tacrolimus concentration was significantly elevated (>30 ng/mL). Both tacrolimus and nefazodone are metabolized by the cytochrome P450 (CYP) 3A4 system. We suspect that nefazodone inhibits tacrolimus metabolism. Therefore, concomitant use of antidepressants such as nefazodone or any other drug that inhibits the CYP3A4 isoenzyme subfamily may interfere with tacrolimus metabolism. Monitoring tacrolimus blood concentrations is crucial, and appropriate dose adjustments are necessary when both drugs are taken concurrently to avoid serious drug interactions. Nephrotoxicity and neurotoxicity are also risks. Depression is a common post-transplant complication and usually requires pharmacological treatment. Many novel selective serotonin reuptake inhibitors (SSRIs) are metabolized by the cytochrome P450 IIIA isoenzyme system, similar to cyclosporine, thus posing an interaction risk to transplant patients. We observed a nearly 10-fold increase in whole blood cyclosporine levels shortly after a heart transplant patient began taking nefazodone as an antidepressant. We believe this is due to a clinically significant drug interaction between nefazodone and cyclosporine, caused by nefazodone's inhibition of the cytochrome P-450 IIIA4 isoenzyme. For more complete data on nefazodone interactions (26 in total), please visit the HSDB record page.

[1]. A review of its pharmacology and clinical efficacy in the management of major depression. Drugs. 1997 Apr;53(4):608-36.

[2]. In vitro assessment of mitochondrial dysfunction and cytotoxicity of nefazodone, trazodone, and buspirone. Toxicol Sci. 2008 Jun;103(2):335-45.

[3]. Effects of nefazodone on the immune system of mice. Eur Neuropsychopharmacol. 2000 Jul;10(4):255-64.

Nefazodone is an N-arylpiperazine, N-alkylpiperazine, triazole, monochlorobenzene, and aromatic ether compound. It possesses pharmacological effects as an antidepressant, serotonergic antagonist, serotonin reuptake inhibitor, alpha-adrenergic antagonist, and analgesic. Nefazodone hydrochloride (trade name: Serzone) is an antidepressant marketed by Bristol-Myers Squibb. Due to the risk of liver damage (although the probability is small), the drug was discontinued in some countries in 2003. Drug-induced liver damage is associated with increased demand for liver transplants and even the risk of death; the incidence of severe liver damage is approximately 1 in 250,000 to 300,000 patient-years. On May 20, 2004, Bristol-Myers Squibb discontinued the sale of Serzone in the United States. Nefazodone is a serotonin reuptake inhibitor. Nefazodone is a serotonergic modulating antidepressant used to treat depression, aggressive behavior, and panic disorder. Nefazodone treatment has been associated with transient, usually asymptomatic elevations in serum transaminase levels and has been linked to several clinically significant cases of acute liver injury, some of which were fatal. Nefazodone hydrochloride (brand name Serzone) is an antidepressant marketed by Bristol-Myers Squibb. Due to the potential (though small) liver damage, which could lead to liver injury requiring transplantation or even death, the drug was discontinued in some countries in 2003. The incidence of severe liver injury is approximately 1 in 250,000 to 300,000 patient-years. On May 20, 2004, Bristol-Myers Squibb discontinued the sale of Serzone in the United States. [Wikipedia] See also: Nefazodone hydrochloride (salt form).

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Drug Indications
For the treatment of depression.

FDA Label

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Mechanism of Action
In the serotonergic system, nefazodone acts as an antagonist of the postsynaptic receptor for serotonin type 2 (5-HT2) and, like fluoxetine antidepressants, inhibits the reuptake of presynaptic serotonin (5-HT). These mechanisms increase the amount of serotonin that can interact with 5-HT receptors. In the noradrenergic system, nefazodone has minimal inhibitory effect on norepinephrine uptake. Nefazodone also antagonizes α1-adrenergic receptors, producing sedative, muscle relaxant, and various cardiovascular effects. Nefazodone has no significant affinity for benzodiazepine, cholinergic, dopaminergic, histaminergic, or β- or α2-adrenergic receptors.

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Therapeutic Use
Second-Generation Antidepressants
/Clinical Trials/ ClinicalTrials.gov is a registry and results database that lists human clinical studies funded by public and private institutions worldwide. The website is maintained by the National Library of Medicine (NLM) and the National Institutes of Health (NIH). Each record on ClinicalTrials.gov provides summary information on the study protocol, including: the disease or condition; the intervention (e.g., the medical product, behavior, or procedure being investigated); the study title, description, and design; participation requirements (eligibility criteria); the location of the study; contact information for the study location; and links to relevant information from other health websites, such as the NLM's MedlinePlus (for patient health information) and PubMed (for citations and abstracts of academic articles in the medical field). Nefazodone is included in the database.
Nefazodone hydrochloride tablets are indicated for the treatment of depression. When choosing an alternative therapy for this condition, prescribing physicians should consider the risk of liver failure associated with nefazodone hydrochloride treatment. In many cases, this will lead doctors to conclude that other medications should be tried first. /US product label contains/

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Drug Warning
/Black Box Warning/ In short-term studies of major depressive disorder (MDD) and other mental illnesses, antidepressants have increased the risk of suicidal ideation and behavior (suicidal tendencies) in children, adolescents, and young adults compared to placebo. Anyone considering the use of nefazodone hydrochloride tablets or any other antidepressant in a child, adolescent, or young adult must weigh this risk against clinical need. Short-term studies have shown that antidepressants do not increase the risk of suicidal tendencies in adults 24 years of age and older compared to placebo; antidepressants reduce the risk of suicidal tendencies in adults 65 years of age and older compared to placebo. Depression and certain other mental illnesses are themselves associated with an increased risk of suicide. Patients of all ages should be appropriately monitored and closely watched for worsening of their condition, suicidal tendencies, or abnormal changes in behavior when starting antidepressant treatment. Family members and caregivers should be informed of the need for close monitoring and communication with the prescribing physician. Nefazodone hydrochloride tablets are not approved for use in children. /Warning/ There have been reports of life-threatening liver failure in patients taking nefazodone hydrochloride tablets. It has been reported that in the United States, approximately 1 in 250,000 to 300,000 patient-years of nefazodone hydrochloride treatment results in death or a liver transplant due to liver failure. Total patient-years refer to the total time each patient receives the medication, expressed in years. For example, one patient-year is equivalent to two patients each receiving six months of treatment, or three patients each receiving four months of treatment, and so on. Generally, nefazodone hydrochloride tablets should not be started in patients with active liver disease or elevated baseline serum transaminases. There is currently no evidence that a history of liver disease increases the risk of liver failure, but abnormal baseline conditions may complicate patient monitoring. Patients should be advised to be aware of signs and symptoms of liver dysfunction (jaundice, anorexia, gastrointestinal discomfort, fatigue, etc.) and to inform their doctor immediately if these symptoms occur. If clinical signs or symptoms suggest liver failure, nefazodone hydrochloride tablets should be discontinued. If a patient develops evidence of hepatocellular injury while taking nefazodone hydrochloride tablets, such as serum AST or serum ALT levels rising to 3 times or more of the upper limit of normal, the medication should be discontinued. If nefazodone hydrochloride is reinstated, an increased risk of liver injury should be assumed in these patients. Therefore, retreatment should not be considered in these patients. The table below lists the most common adverse events (incidence ≥5%) following nefazodone use, which occurred at a significantly higher rate in the placebo group than in the placebo group (i.e., significantly higher in the nefazodone group than in the placebo group, p≤0.05), including: somnolence, dry mouth, nausea, dizziness, constipation, fatigue, lightheadedness, blurred vision, confusion, and visual disturbances. In a global premarketing clinical trial, approximately 16% of the 3496 patients treated with nefazodone discontinued treatment due to adverse reactions. In clinical trials, more common (≥1%) and considered drug-related events associated with discontinuation (i.e., the dropout rate in the nefazodone group was approximately twice or more than that in the placebo group) included: nausea (3.5%), dizziness (1.9%), insomnia (1.5%), fatigue (1.3%), and agitation (1.2%). For more complete (17) drug warnings for nefazodone, please visit the HSDB record page. Pharmacodynamics: Nefazodone is a synthetic antidepressant derived from phenylpiperazine compounds and used to treat major depressive disorder. Although nefazodone has a structure similar to trazodone, its mechanism of action differs from other antidepressants. Therefore, it does not carry the severe cardiovascular toxicity risks associated with tricyclic antidepressants, nor does it cause insomnia and REM sleep suppression like selective serotonin reuptake inhibitors.

C25H32N5O2CL

470.00688

469.224

C, 63.89; H, 6.86; Cl, 7.54; N, 14.90; O, 6.81

83366-66-9

Nefazodone hydrochloride;82752-99-6

4449

Light yellow to yellow solid powder

180-182°C

3.554

0

5

10

33

649

0

CCC1=NN(CCCN2CCN(CC2)C2=CC(Cl)=CC=C2)C(=O)N1CCOC1=CC=CC=C1

VRBKIVRKKCLPHA-UHFFFAOYSA-N

InChI=1S/C25H32ClN5O2/c1-2-24-27-31(25(32)30(24)18-19-33-23-10-4-3-5-11-23)13-7-12-28-14-16-29(17-15-28)22-9-6-8-21(26)20-22/h3-6,8-11,20H,2,7,12-19H2,1H3

2-[3-[4-(3-chlorophenyl)piperazin-1-yl]propyl]-5-ethyl-4-(2-phenoxyethyl)-1,2,4-triazol-3-one

Nefazodone, Dutonin, Nefadar, Serzone

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

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

DMSO : ~125 mg/mL (~265.95 mM)

Solubility in Formulation 1: ≥ 2.08 mg/mL (4.43 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 20.8 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 2: ≥ 2.08 mg/mL (4.43 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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