JNK;MMP13

The HIF-1α pathway in NSCLC cells is inhibited by chloramphenicol (1-100 μg/mL, 18-24 h) in a concentration-dependent manner.NSCLC cells are exposed to 100 μg/mL of chloramphenicol for 0–24 hours, which causes autophagy induction and significantly raises the levels of autophagic biomarkers (beclin-1, Atg12–Atg5 conjugates, and LC3–II)[1].
In activated T cells, chloramphenicol inhibits apoptosis and causes aberrant differentiation[2].
Chloramphenicol can cause reduced ATP biosynthesis and mitochondrial stress by blocking the synthesis of proteins in both bacteria and mitochondria[3].
Chloramphenicol (1-100 μg/mL) has the ability to upregulate MMP-13 protein and stimulate the expression of matrix metalloproteinase (MMP)-13[3].
Chloramphenicol (1-100 μg/mL) has the ability to stimulate PI-3K/Akt signaling, c-Jun protein phosphorylation, and c-Jun N-terminal kinases (JNK)[3].
By inhibiting peptidyl transferase activity, chloramphenicol mainly affects the 50S subunit of bacterial 70S rihosomes, which prevents the formation of peptide bonds[5].

Day 1 post-dosing sees a decrease in marrow erythroid cells and erythrocyte precursors, and by day 14, after 14 days of treatment, erythrocytes and erythrocyte precursors have returned to normal[4].

Animal Model: Female B6C3F1 mice (12-14 weeks old)
Dosage: 0, 2500 and 3500 mg/kg
Administration: Gavage, daily, for 5 days
Result: On the first day after dosage, erythropoiesis was clearly stopped. At the 2500 mg/kg dose level on day 7 and between 7 and 14 days at the 3500 mg/kg dose level, respectively, a recovery was observed after the dosage. At every dosage level, the erythroid series showed the greatest myelotoxicity. Day 1 post-dosage: decreased femoral marrow BFU-E and CFU-E. Within 14 days of the dosage, every blood and marrow parameter in the current study was back to normal.

Absorption, Distribution and Excretion
After oral administration, it is rapidly and completely absorbed from the gastrointestinal tract (bioavailability 80%). Intramuscular injection results in good absorption (bioavailability 70%). Topical application to the eyes also results in intraocular absorption and partial systemic absorption. It is primarily eliminated by hepatic metabolism to inactive glucuronide. This metabolite, along with chloramphenicol itself, is excreted in the urine after filtration and secretion. Oral chloramphenicol is rapidly absorbed from the intestine. In controlled studies in adult volunteers, the recommended dose of 50 mg/kg/day, 1 g every 6 hours for 8 doses, was used. Using a microbiological assay, the mean peak serum concentration 1 hour after the first dose was 11.2 μg/mL. The cumulative effect resulted in a peak plasma concentration of 18.4 μg/mL after the fifth 1 g dose. The mean plasma concentration over 48 hours ranged from 8 to 14 μg/mL. In these studies, the total urinary excretion rate of chloramphenicol over three days ranged from a minimum of 68% to a maximum of 99%. Of the excreted antibiotics, 8% to 12% exist as free chloramphenicol; the remainder are inactive microbial metabolites, primarily products conjugated with glucuronic acid. Due to the rapid excretion of glucuronides, most chloramphenicol detected in the blood exists in its active, free form. Although the proportion of the unchanged drug excreted in urine is small, the concentration of free chloramphenicol is relatively high, reaching hundreds of micrograms per milliliter in patients receiving 50 mg/kg/day in divided doses. Small amounts of active drug can be detected in bile and feces. Chloramphenicol diffuses rapidly but unevenly. The highest concentrations are found in the liver and kidneys, and the lowest in brain tissue and cerebrospinal fluid. Even without meningitis, chloramphenicol can enter the cerebrospinal fluid at concentrations approximately half that in the blood. Measurable drug concentrations can also be detected in pleural effusion, ascites, saliva, breast milk, as well as aqueous humor and vitreous humor. Chloramphenicol is transported across the placental barrier, with concentrations in neonatal umbilical cord blood slightly lower than in maternal blood. Following oral, intravenous, and intraperitoneal administration, chloramphenicol rapidly reaches peak serum concentrations. Intramuscular administration of chloramphenicol, except for certain soluble formulations, results in slightly delayed absorption and lower serum concentrations compared to oral, intravenous, or intraperitoneal routes. Chloramphenicol readily diffuses into all body tissues, though at varying concentrations. The highest concentrations are found in the liver and kidneys of dogs, indicating these organs are the primary pathways for the inactivation and excretion of its metabolites. Chloramphenicol concentrations in the lungs, spleen, heart, and skeletal muscle are similar to those in the blood. Chloramphenicol diffuses from the blood into the aqueous humor and vitreous humor, where it reaches significant concentrations. A significant difference from other antibiotics is its extremely strong cerebrospinal fluid diffusion capacity. Within three to four hours of administration, the average concentration of chloramphenicol in the cerebrospinal fluid can reach 50% of the serum concentration. This percentage may be even higher if the meninges are inflamed. Chloramphenicol readily diffuses into milk, pleural fluid, and ascites, and can cross the placenta, reaching concentrations up to approximately 75% of the maternal blood concentration. Approximately 55% of the daily single dose is recovered from the urine of treated dogs. A small fraction of this remains as unmetabolized chloramphenicol. For more complete data on absorption, distribution, and excretion of chloramphenicol (21 in total), please visit the HSDB record page. Metabolism/Metabolites Primarily metabolized in the liver, 90% is bound to inactive glucuronic acid. Chloramphenicol is rapidly metabolized, primarily in the liver by binding to glucuronic acid. In humans, it forms d-threo-2-amino-1-(p-nitrophenyl)-1,3-propanediol and chloramphenicol-β-D-glucuronide. In rats. /Excerpt from table/.../occurs/direct binding. The formation of glucuronide has been shown to occur on primary alcohols rather than secondary alcohols… its primary function is to inactivate and detoxify the drug in the human body; any factor that reduces its effect… will lead to a significant increase in toxicity. ...In newborns...bilirubin...acts as a competitive endogenous receptor.
Chloramphenicol 3-glucuronide is the major metabolite of chloramphenicol produced by isolated rat hepatocytes, although a minor metabolite is also produced.
For more complete metabolite/metabolite data on chloramphenicol (10 in total), please visit the HSDB record page.
Biological half-life
The half-life in adults with normal liver and kidney function is 1.5–3.5 hours. The half-life in patients with impaired kidney function is 3–4 hours. The half-life in patients with severe liver impairment is 4.6–11.6 hours. The half-life in children aged 1 month to 16 years is 3–6.5 hours, while the half-life in infants aged 1–2 days is 24 hours or longer, with significant individual variability, especially in low birth weight infants. The half-life of chloramphenicol in humans ranges from 1.6 to 4.6 hours; the apparent volume of distribution, measured using different techniques in various adult patients, ranges from 0.2 to 3.1 liters/kg… The half-life is significantly prolonged in newborns… the half-life in infants aged 1–8 days is 10–48 hours or more, and in infants aged 11 days to 8 weeks is 5–16 hours… In adults with normal renal and hepatic function, the plasma half-life of chloramphenicol is 1.5–4.1 hours… The plasma half-life in infants aged 1–2 days is 24 hours or longer, and in infants aged 10–16 days is approximately 10 hours. Patients with significantly impaired hepatic function have a prolonged plasma half-life of chloramphenicol. Patients with impaired renal function do not have a significantly prolonged plasma half-life of chloramphenicol, but the half-life of its inactive conjugates may be prolonged.

Toxicity Summary
Identification and Uses: Chloramphenicol is an antibiotic used in humans and veterinary medicine. It was originally isolated from the soil bacterium Streptomyces venzeuelae. Human Studies: Chloramphenicol is known to cause serious side effects in humans. One of these is usually irreversible aplastic anemia and reversible bone marrow suppression. An association between chloramphenicol and liver disease has been found. Acute myeloid leukemia has also been associated with chloramphenicol exposure. Newborns, especially premature infants, may develop a serious condition called "gray baby syndrome" if exposed to excessive amounts of chloramphenicol. This syndrome typically appears 2–9 days (average 4 days) after the start of treatment. Within the first 24 hours, the infant may experience vomiting, refusal to breathe, irregular and rapid breathing, abdominal distension, intermittent cyanosis, and loose green stools. The infant becomes critically ill by the end of the first day and develops pallor, hypotonia, and hypothermia over the next 24 hours. Similar "gray baby syndrome" has been reported in adults after accidental overdose of the drug. A patient who used a chloramphenicol-containing topical cream developed allergic contact dermatitis. Additionally, there have been reports of optic neuritis and visual impairment due to scotomas. Chloramphenicol is associated with cleft lip. In vitro experiments have shown that chloramphenicol-treated human lymphocytes exhibit chromosomal abnormalities. Animal experiments: Ten 3-month-old mice in three groups were intraperitoneally injected daily with chloramphenicol at doses of 20, 40, or 100 mg/kg body weight for three months. Results showed that the mice developed splenomegaly, hepatomegaly, lymphadenopathy, and thymic hypertrophy in a dose-dependent manner. The addition of chloramphenicol to drinking water increased the incidence of lymphoma in both mouse strains. Chloramphenicol has been shown to cause DNA strand breaks in bacterial cells and inhibit DNA synthesis in lymphocytes and E. coli bacteriophages. Chromosomal abnormalities were observed in bone marrow cells from mice that received intraperitoneal or intramuscular injections of chloramphenicol in the livers of F1 generation mice. Ototoxicity was observed in rats when 3–9 female Sprague-Dawley rats were divided into several groups and chloramphenicol was added to their drinking water, with or without short-term high-intensity noise stimulation. Oral administration of this antibiotic inhibited REM sleep in cats. Chloramphenicol administration to pregnant monkeys had no effect on fetal development. Oral administration of high doses of chloramphenicol (500-2000 mg/kg) to rats and mice, and to rabbits (500 and 1000 mg/kg) to rabbits, resulted in high embryonic and fetal mortality rates and fetal growth retardation in all three animal groups. Teratogenic effects (primarily manifested as umbilical hernia) were observed only in rats. No other toxic symptoms were observed in pregnant animals except for significantly lower body weight gain in the highest dose group compared to the control group. Offspring of chloramphenicol-treated mice showed decreased learning ability, increased brain epilepsy thresholds, and poor performance in open field tests. Ecotoxicity studies: Chloramphenicol was added to cultures of one freshwater green alga, Chlorella pyrenoidosa, and two marine algae, Isochrysis galbana and Tetraselmis chui. Results showed that chloramphenicol was more toxic to freshwater green algae. Chloramphenicol exposure significantly inhibited the growth of Scenedesmus obliquus, while Chlorella showed lower sensitivity. African catfish (Clarias gariepinus) exposed to chloramphenicol exhibited abnormal behavioral changes, and hematological parameters were also affected. In Egyptian toads (Bufo regularis), administration of 5 mg chloramphenicol per 40 g body weight for 12 consecutive weeks resulted in significant ultrastructural alterations in almost all types of leukocytes. Hepatotoxicity
Some patients with hematologic disorders caused by chloramphenicol also experienced clinically significant liver damage, accompanied by jaundice, usually prior to the onset of aplastic anemia or severe thrombocytopenia. Jaundice occurs in 10% to 25% of aplastic anemia cases, typically within 1 to 2 months of initiation of chloramphenicol treatment, and often shortly after discontinuation. Aplastic anemia and its associated liver damage most commonly occur in patients receiving multiple courses of chloramphenicol or undergoing long-term treatment. Serum enzyme profiles are typically hepatocellular, clinically presenting as an acute hepatitis-like syndrome, with onset of fatigue, nausea, anorexia, and abdominal discomfort, followed by darkening of urine and jaundice. A few cases present with a cholestatic pattern, accompanied by jaundice, pruritus, and significantly elevated alkaline phosphatase. Some cases do not involve bone marrow. Immune hypersensitivity and autoimmune features are rare. Most cases resolve spontaneously, but acute liver failure has been reported, especially in patients without aplastic anemia. However, in most cases, chloramphenicol-induced liver damage is masked by severe bone marrow aplasia. Probability Score: B (Very likely a cause of clinically apparent liver damage, but now rare). Effects during Pregnancy and Lactation ◉ Overview of Use During Lactation Infants breastfed by mothers taking oral chloramphenicol have been reported to experience adverse reactions such as vomiting, flatulence, and drowsiness during breastfeeding. The concentration of chloramphenicol in breast milk is insufficient to induce "gray baby syndrome," but since chloramphenicol-induced aplastic anemia is dose-independent, this could occur, although no reports have been made. During breastfeeding, especially with newborns or premature infants, it is best to choose other medications instead of chloramphenicol. If the mother must take chloramphenicol while breastfeeding, the infant's gastrointestinal reactions and breastfeeding should be monitored. It is recommended to monitor the infant's complete blood count and differential blood count. In some cases, it may be necessary to discontinue breastfeeding.
◉ Effects on Breastfed Infants
A study reported on 50 breastfed infants whose mothers began oral chloramphenicol 2 to 12 days postpartum at doses of 1 gram (n=20), 2 grams (n=20), or 3 grams (n=10) daily. All infants refused to suckle, and 50% to 60% fell asleep during breastfeeding. Among the infants whose mothers took 1 gram, 2 grams, and 3 grams daily, 10%, 25%, and 90%, respectively, vomited after breastfeeding. All infants experienced flatulence and bloating, with serious problems occurring in 0.5%, 20%, and 100% of infants whose mothers administered 1 gram, 2 grams, and 3 grams daily, respectively.
◉ Effects on Lactation and Breast Milk
No published information found as of the revision date.
Protein Binding

Adult plasma protein binding is 50-60%, and 32% in preterm newborns.
Drug Interactions

Because chloramphenicol interferes with erythrocyte maturation, patients receiving chloramphenicol may respond poorly to cyanocobalamin treatment.
Concomitant use with other drugs that may cause bone marrow suppression should be avoided.
Chloramphenicol antagonizes the effects of antibiotics such as penicillin and streptomycin, which act only on growing cells, but it has a synergistic effect with tetracycline, which also works by inhibiting protein synthesis. Chloramphenicol may produce similar synergistic effects with other antibiotics that work by inhibiting protein synthesis. Human clinical observations and corresponding experimental animal studies (infected with various pathogens) have shown that the combined use of chloramphenicol with gamma globulin or specific antiserum has a greater therapeutic effect than the combined effect of the two alone. For more complete data on chloramphenicol interactions (26 in total), please visit the HSDB record page. Non-human toxicity values: Rat intraperitoneal LD50: 1811 mg/kg
Rat subcutaneous LD50: 5 g/kg
Rat intravenous LD50: 171 mg/kg
Mouse oral LD50: 1500 mg/kg
For more complete data on chloramphenicol non-human toxicity values (14 in total), please visit the HSDB record page.

[1]. Chloramphenicol Induces Autophagy and Inhibits the Hypoxia Inducible Factor-1 Alpha Pathway in Non-Small Cell Lung Cancer Cells. Int J Mol Sci. 2019 Jan 3;20(1):157.

[2]. Chloramphenicol induces abnormal differentiation and inhibits apoptosis in activated T cells. Cancer Res. 2008 Jun 15;68(12):4875-81.

[3]. Chloramphenicol causes mitochondrial stress, decreases ATP biosynthesis, induces matrix metalloproteinase-13 expression, and solid-tumor cell invasion. Toxicol Sci. 2010 Jul;116(1):140-50.

[4]. Characterization of the myelotoxicity of chloramphenicol succinate in the B6C3F1 mouse. Int J Exp Pathol. 2006 Apr;87(2):101-12.

[5]. Jardetzky, O., Studies on the mechanism of action of chloramphenicol. I. The conformation of chlioramphenicol in solution. J Biol Chem, 1963. 238: p. 2498-508.

[6]. Wolfe, A.D. and F.E. Hahn, Mode of Action of Chloramphenicol. Ix. Effects of Chloramphenicol Upon a Ribosomal Amino Acid Polymerization System and Its Binding to Bacterial Ribosome. Biochim Biophys Acta, 1965. 95: p. 146-55

Therapeutic Uses
Antimicrobial Agent; Protein Synthesis Inhibitor
/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 includes summary information about the study protocol, including: disease or condition; intervention (e.g., the medical product, behavior, or procedure being investigated); the title, description, and design of the study; 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 providing patient health information) and PubMed (for providing citations and abstracts of academic articles in the medical field). Chloramphenicol is listed in the database.
Chloramphenicol is an antibiotic produced by Streptomyces venezuelae…recommended for serious infections where the site of infection, pathogen susceptibility, or poor response to other therapies indicates limited antimicrobial options. Since the 1950s, it has been used to treat a variety of microbial infections, including typhoid fever and other forms of salmonellosis, as well as central nervous system, anaerobic, and eye infections… (Veterinarian): Chloramphenicol tablets are recommended for oral treatment of the following diseases in dogs: bacterial lung infections caused by susceptible microorganisms, such as Staphylococcus aureus, Streptococcus pyogenes, and Brucella; urinary tract infections caused by susceptible microorganisms such as Escherichia coli, Proteus vulgaris, Corynebacterium nephritis, Streptococcus spp., and hemolytic Staphylococcus; enteritis caused by susceptible microorganisms such as Escherichia coli, Proteus spp., Salmonella spp., and Pseudomonas spp.; and canine distemper-related infections caused by susceptible microorganisms such as Bordetella bronchiseptica, Escherichia coli, Pseudomonas aeruginosa, Proteus spp., Shigella spp., and Moraxella catarrhalis. Adjunctive therapy should be used when necessary. Most susceptible pathogens respond to chloramphenicol treatment for 3 to 5 days at the recommended dosage regimen. If no therapeutic effect is observed after 3 to 5 days, use should be discontinued and the diagnosis reassessed. Furthermore, a change in treatment regimen should be considered. Laboratory tests, including in vitro culture and drug sensitivity testing, should be performed on samples collected before treatment. For more complete data on the therapeutic uses of chloramphenicol (38 types in total), please visit the HSDB record page.
Drug Warnings
/Black Box Warning/ Warning: Serious and potentially fatal blood disorders (aplastic anemia, hypoplastic anemia, thrombocytopenia, and granulocytopenia) are known to occur with chloramphenicol. In addition, there have been reports of chloramphenicol-induced aplastic anemia eventually developing into leukemia. Blood disorders may occur with both short-term and long-term use of this drug. Chloramphenicol should not be used when other potentially less risky medications are effective, as described in the "Indications and Usage" section. This product should not be used to treat minor infections or inappropriate conditions, such as colds, influenza, or throat infections; nor should it be used as a prophylactic agent for bacterial infections. Precautions: Adequate blood tests must be performed during treatment. While blood tests can detect early peripheral blood changes, such as leukopenia, reticulopenia, or granulocytopenia, thus preventing irreversible progression of the disease, they cannot be relied upon to detect bone marrow suppression prior to the onset of aplastic anemia. Hospitalization is recommended to facilitate appropriate examination and observation during treatment.
/Black Box Warning/ Warning: There have been reports of bone marrow dysplasia (including aplastic anemia) and death following topical application of chloramphenicol. Chloramphenicol should not be used when effective treatment with a potentially less risky medication is expected.
Chloramphenicol is not recommended for routine treatment of typhoid carrier states.
Chloramphenicol is contraindicated in patients with a history of hypersensitivity and/or toxicity to chloramphenicol. It should not be used to treat minor infections or inappropriate conditions, such as colds, influenza, or throat infections; nor should it be used as a prophylactic agent for bacterial infections.
For more complete data on chloramphenicol warnings (44 in total), please visit the HSDB records page.

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Pharmacodynamics
Chloramphenicol is a broad-spectrum antibiotic originally derived from Streptomyces venezulatus and now produced synthetically. Chloramphenicol is effective against a wide range of microorganisms, but due to its serious side effects in humans (e.g., bone marrow damage, including aplastic anemia), it is typically used only to treat severe, life-threatening infections (e.g., typhoid fever). Chloramphenicol is a bacteriostatic agent, but may have bactericidal effects at high concentrations or when used to treat highly susceptible microorganisms. Chloramphenicol inhibits bacterial growth by binding to bacterial ribosomes (blocking peptidyl transferases) and inhibiting protein synthesis.

C11H12CL2N2O5

323.13

322.012

C, 40.89; H, 3.74; Cl, 21.94; N, 8.67; O, 24.76

56-75-7

Chloramphenicol-d5;202480-68-0;Chloramphenicol palmitate;530-43-8;Levomecol;118573-58-3;DL-threo-Chloramphenicol-d5;1420043-66-8;Threo-Chloramphenicol-d6;Chloramphenicol-d4

5959

White to off-white crystalline powder.

1.6±0.1 g/cm3

563.2±60.0 °C at 760 mmHg

148-150 °C(lit.)

294.4±32.9 °C

0.0±1.6 mmHg at 25°C

1.623

1.62

3

5

5

20

342

2

ClC([H])(C(N([H])[C@]([H])(C([H])([H])O[H])[C@@]([H])(C1C([H])=C([H])C(=C([H])C=1[H])[N+](=O)[O-])O[H])=O)Cl

WIIZWVCIJKGZOK-RKDXNWHRSA-N

InChI=1S/C11H12Cl2N2O5/c12-10(13)11(18)14-8(5-16)9(17)6-1-3-7(4-2-6)15(19)20/h1-4,8-10,16-17H,5H2,(H,14,18)/t8-,9-/m1/s1

2,2-dichloro-N-[(1R,2R)-1,3-dihydroxy-1-(4-nitrophenyl)propan-2-yl]acetamide

Chloramphenicol; Chlornitromycin; Chloromycetin; Levomycetin; Chlorocid; Globenicol; Detreomycin; Kloramfenikol; Levomycetin; Ophthochlor; Syntomycin;

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: This product is not stable in solution, please use freshly prepared working solution for optimal results.

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

DMSO :65~150 mg/mL (201.15~464.21 mM )
Ethanol : ~100 mg/mL (~309.47 mM )
H2O : ~3.06 mg/mL (~9.47 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (7.74 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.

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Solubility in Formulation 2: ≥ 2.5 mg/mL (7.74 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.

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Solubility in Formulation 3: ≥ 2.5 mg/mL (7.74 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.

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Solubility in Formulation 4: ≥ 2.5 mg/mL (7.74 mM) (saturation unknown) in 10% EtOH + 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 EtOH stock solution to 400 μL of PEG300 and mix evenly; then add 50 μL of Tween-80 to the above solution and mix evenly; then add 450 μL of 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 5: ≥ 2.5 mg/mL (7.74 mM) (saturation unknown) 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.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 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.

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Solubility in Formulation 6: ≥ 2.5 mg/mL (7.74 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 25.0 mg/mL clear EtOH stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 7: 10% DMSO+40% PEG300+5% Tween-80+45% Saline: ≥ 2.5 mg/mL (7.74 mM)

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Solubility in Formulation 8: 2.5 mg/mL (7.74 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication.

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