Absorption, Distribution and Excretion
The oral bioavailability of cyclobenzafir is estimated to be between 0.33 and 0.55. The Cmax is 5–35 ng/mL, and the time to peak concentration (Tmax) is 4 hours. The AUC over an 8-hour dosing interval has been reported to be approximately 177 ng·hr/mL. Following administration of radiolabeled cyclobenzafir, 38–51% of the radioactive material is excreted in the urine, and 14–15% in the feces. Cyclobenzafir is rapidly metabolized, with only about 1% of the radiolabeled dose recovered unchanged in the urine. The metabolites excreted in the urine are likely water-soluble glucuronide conjugates. The volume of distribution of cyclobenzafir is approximately 146 liters. Cyclobenzafir has a relatively long half-life, but a high plasma clearance rate, suggesting its widespread tissue distribution. The plasma clearance of cyclobenzafir is approximately 0.7 L/min. Cyclobenzafir is widely distributed throughout the body. ...It is unclear whether cyclobenzaline can cross the placenta. The drug binds well to plasma proteins (approximately 93%). /Breast Milk/ It is unclear whether cyclobenzaline is distributed into human breast milk; however, it is distributed into breast milk in rats.
This study investigated the absorption, distribution, excretion, and metabolism of 3-(5H-dibenzo[a,d]cycloheptene-5-ylidene)-N,N-dimethyl-1-propane (cyclobenzaline) in rats, dogs, rhesus monkeys, and humans. The drug was well absorbed in all animals after oral administration. Rats excreted the drug primarily via feces, while dogs, monkeys, and humans excreted it primarily via urine. The drug was rapidly and extensively distributed in various tissues in rats, with the highest concentrations in the small intestine, lungs, kidneys, and liver. The drug binds well to human plasma. Significant bile excretion of the labeled compound was observed in rats. The main metabolites in rats are phenolic derivatives, while in humans the main metabolites are 10,11-dihydroxynortriptyline and cyclobenzafyl glucuronide. Only trace amounts of unmetabolized drug were detected in urine. Cyclobenzafyl is well absorbed orally. It circulates enterohepaticly and appears to be metabolized on its first pass through the gastrointestinal tract and/or liver. The mean oral bioavailability of this drug is estimated to be 33-55%. Following a single oral dose of 5 mg or 10 mg cyclobenzafyl hydrochloride, peak plasma concentrations of 4.3 ng/mL and 8.5 ng/mL, respectively, are reached approximately 4 hours later. With three daily doses of cyclobenzafyl, steady-state plasma concentrations are reached within 3-4 days, approximately four times higher than those following a single dose. In healthy individuals, with three daily doses of the drug, mean steady-state peak plasma cyclobenzafyl concentrations of 14.9 ng/mL and 25.9 ng/mL were reached 4 hours and 3.9 hours, respectively, following doses of 5 mg and 10 mg.

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Metabolism/Metabolites
Cyclobenzaline is extensively metabolized in the liver via oxidative and conjugative pathways. Oxidative metabolism, primarily N-demethylation, is mainly catalyzed by CYP3A4 and CYP1A2 (with less involvement of CYP2D6), producing the major metabolite demethylcyclobenzaline. Cyclobenzaline undergoes N-glucuronidation in the liver catalyzed by UGT1A4 and UGT2B10, and has been shown to circulate enterohepaticly. In the urine of dogs orally administered the labeled drug, ten cyclobenzaline metabolites were identified, accounting for approximately 50% of urinary radioactivity. These metabolites include 1,2-dihydrodiol, three phenolic derivatives, N-oxide, 10,11-epoxide, 10,11-ethylene glycol, demethylcyclobenzaline, and demethylcyclobenzaline and cyclobenzaline glucuronide conjugates. These metabolites are excreted in both free and conjugated forms. Only small amounts of unmetabolized cyclobenzaline are present. Cyclobenzaline is widely metabolized, primarily excreted via the kidneys as a glucuronide. Cytochrome P-450 isoenzymes 3A4, 1A2, and (lessly) 2D6 mediate N-demethylation, one of the oxidative pathways of cyclobenzaline. This study investigated the absorption, distribution, excretion, and metabolism of 3-(5H-dibenzo[a,d]cyclohepten-5-ylidene)-N,N-dimethyl-1-propylamine (cyclobenzaline) in rats, dogs, rhesus monkeys, and humans. …The main metabolites in rats are phenolic derivatives, while in humans, the main metabolites are 10,11-dihydroxynortriptyline and cyclobenzaline glucuronide. Cyclobenzaline is primarily metabolized through oxidative and conjugative pathways. Hepatic cytochrome P-450 (CYP) isoenzymes 3A4, 1A2, and (lessly) 2D6 are responsible for the oxidative N-demethylation of this drug. Orally administered cyclobenzapine is primarily excreted in the urine as an inactive glucuronide metabolite; less than 1% of the drug is excreted unchanged via the kidneys. The fungus Cunninghamella elegans was used as a microbial model for drug metabolism in mammals to biotransform the tricyclic antidepressant cyclobenzapine. Cunninghamella elegans cultured in Sabouraud broth metabolized 75% of the drug at a concentration of 1 mM within 72 hours. Milligram-level fungal metabolites were isolated using reversed-phase high-performance liquid chromatography (HPLC), and their structures were characterized by 1H NMR, mass spectrometry, and ultraviolet spectroscopy. The major fungal metabolites of cyclobenzazarine include 2-hydroxycyclobenzazarine (59%), N-demethylcyclobenzazarine (21%), cyclobenzazarine trans-10,11-dihydrodiol (5%), N-demethyl-2-hydroxycyclobenzazarine (3%), 3-hydroxycyclobenzazarine (3%), and cyclobenzazarine N-oxide (1%). These fungal metabolites were used as standards to study the metabolism of cyclobenzazarine in rat liver microsomes. Rat liver microsomes can also biotransform cyclobenzazarine, producing metabolites similar to those of the fungi. Isotopic labeling of 2-hydroxycyclobenzazarine with 18O2 and the trans configuration of the dihydrodiol indicate that these reactions are catalyzed by cytochrome P-450 monooxygenase from C. elegans. These results also suggest that fungal biotransformation systems can be used to predict and synthesize drug metabolites in mammals. Known metabolites of cyclobenzazarine include N-demethylcyclobenzazarine.

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Biological Half-Life
The effective half-life of cyclobenzapine in young, healthy subjects is approximately 18 hours. These values are prolonged in older adults and patients with hepatic impairment, with mean effective half-lives of 33.4 hours and 46.2 hours, respectively.
Cyclobenzapine is eliminated rather slowly, with an effective half-life of 18 hours (range 8–37 hours; n=18)...

Toxicity Summary
Identification and Use: Cyclobenzafine (used in cyclobenzafine hydrochloride tablets) is indicated as an adjunct to rest and physical therapy to relieve muscle spasms caused by acute painful musculoskeletal disorders. Human Exposure and Toxicity: Overdose of cyclobenzafine can lead to rapid onset of toxicity, and in rare cases, death. The most common toxic reactions to cyclobenzafine overdose are drowsiness and tachycardia; less common reactions include tremor, agitation, coma, ataxia, hypertension, slurred speech, confusion, dizziness, nausea, vomiting, and hallucinations. In rare cases, serious adverse reactions may occur, including cardiac arrest, chest pain, arrhythmias, severe hypotension, seizures, and neuroleptic malignant syndrome. Serotonin syndrome is another potential side effect. Cyclobenzafine blood concentrations ≥0.8 mg/L can be fatal. In one case of accidental overdose, the patient experienced severe hypothermia and cardiac arrest during transport. Animal studies: In dogs, a single oral gavage administration of 180 mg/kg or higher doses of cyclobenzarline resulted in salivation, vomiting, tremors, convulsions, and increased respiratory rate, leading to death within one hour. Acute exposure in rats resulted in ataxia, decreased respiratory rate, sedation, hind limb flaccidity, loss of ear reflexes, loss of righting reflex during swimming, intermittent clonic convulsions, weight loss, lethargy, and ultimately death. The drug is more toxic to young and weaned rats than to adult rats. In rats treated with cyclobenzarline hydrochloride for up to 67 weeks at doses approximately 5 to 40 times the maximum recommended human dose, pale livers, sometimes enlarged, and dose-related hepatocyte vacuolation and lipid degeneration were observed. In mouse and rabbit studies, no evidence of embryonal lethality or teratogenicity was found after oral administration of doses of 5, 10, or 20 mg/kg/day. In rats, doses of 5 or 10 mg/kg/day had no adverse effects on male and female reproductive capacity and fertility, or on the growth and survival of their offspring. A dose of 20 mg/kg/day reduced litter size, pup size and survival, and increased maternal weight. Cyclobenzaline hydrochloride did not show genotoxic effects in multiple studies, including the mouse bone marrow micronucleus test. Salmonella-Escherichia coli mammalian microsomal reverse mutation assay and confirmatory test; and chromosomal aberration assay with/without metabolic activation in CHO cells.

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Hepatotoxicity
The cyclobenzaline product information states that abnormal liver function, hepatitis, jaundice, and cholestasis may occur in the following situations:
Probability score: E (unproven but suspected cause of clinically significant liver injury).

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Effects during pregnancy and lactation
◉ Overview of use during lactation
Cyclobenzaline appears to be present in very low concentrations in breast milk, and both infants appeared to tolerate the drug well in their breast milk. If the mother needs to take cyclobenzalin, breastfeeding does not need to be stopped. Monitor the infant for lethargy, weight gain, and developmental milestones, especially in newborns and preterm infants, and when using multiple sedatives.
◉ Effects on breastfed infants
Two mothers were on long-term cyclobenzalin use. One mother took 5 mg daily for temporomandibular joint pain; the other took 10 mg daily for fibromyalgia. The latter mother also took an unspecified antidepressant, levothyroxine, zolpidem, alprazolam, and famotidine. Both mothers were breastfeeding (the extent of breastfeeding was not specified). Neither infant experienced any significant adverse reactions, such as sedation.
We searched a shared database of all poison control centers in the United States for consultation calls related to medications and breastfeeding between 2001 and 2017. Of the 2,319 cases of infants exposed to a substance through breast milk, 1 of the 7 cases classified as causing serious adverse reactions involved cyclobenzalin. A 16-day-old infant was exposed to cyclobenzafine, acetaminophen, and oxycodone via breast milk. The infant was admitted to the non-intensive care unit of a hospital due to bradycardia, hypotension, and respiratory arrest. The dosage and extent of breastfeeding were not reported, and the infant ultimately survived.
◉ Effects on breastfeeding and breast milk
As of the revision date, no relevant published information was found.

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Protein binding
Cyclobenzafine has a protein binding rate of approximately 93% in plasma. It has been shown to have a high affinity for human serum albumin.

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Drug interactions
Cyclobenzafine has been reported to be well tolerated in combination with diflunisal or naproxen without any unexpected adverse reactions. However, combination of cyclobenzafine and naproxen increases the incidence of drowsiness. Plasma concentrations are not affected when aspirin and cyclobenzafine are used together. It has not been determined whether combination therapy with aspirin (or other analgesics) improves clinical efficacy. Cyclobenzalin and structurally similar tricyclic antidepressants may block the antihypertensive effects of guanethidine (discontinued in the US) and other similar drugs. Cyclobenzalin and structurally similar tricyclic antidepressants may increase the risk of seizures in patients taking tramadol. Cyclobenzalin may have additive or synergistic effects with other central nervous system depressants (e.g., alcohol, barbiturates). Cyclobenzalin, especially when used in combination with alcohol or other central nervous system depressants, may impair a patient's ability to perform activities requiring mental alertness or physical coordination (e.g., operating machinery, driving a motor vehicle). For more complete data on cyclobenzalin interactions (9 in total), please visit the HSDB record page.

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Non-human toxicity values
Mouse intravenous LD50: 36 mg/kg
Mouse intraperitoneal LD50: 90 mg/kg
Mouse oral LD50: 250 mg/kg
Rat oral LD50: 425 mg/kg
Mouse oral LD50: 338 mg/kg

Eur J Pharmacol.1996 Sep 5;311(1):29-35;Eur J Pharmacol.2003 Jan 1;458(1-2):91-9.

Therapeutic Uses

Tricyclic antidepressants; central muscle relaxants; sedatives
/Clinical Trials/ ClinicalTrials.gov is a registry and results database that indexes 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 a summary of the study protocol, including: the disease or condition; the intervention (e.g., the medical product, behavior, or procedure under investigation); 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 patient health information) and PubMed (for citations and abstracts of academic articles in the medical field). Cyclobenzapine is indexed in the database.
Cyclobenzapine hydrochloride tablets are indicated as an adjunct to rest and physical therapy for the relief of muscle spasms caused by acute painful musculoskeletal disorders. Its efficacy is reflected in the relief of muscle spasms and related signs and symptoms, such as pain, tenderness, limited mobility, and limited activities of daily living. Cyclobenzapine hydrochloride should only be used for short periods (maximum two to three weeks) because there is insufficient evidence to suggest its long-term effectiveness, and muscle spasms caused by acute painful musculoskeletal disorders are usually short-lived and rarely require long-term specific treatment. Cyclobenzapine hydrochloride is ineffective for spasms caused by brain or spinal cord diseases, or for spasms in children with cerebral palsy. /US product label includes/
Some data suggest that cyclobenzapine may be effective in treating fibromyalgia. Cyclobenzapine is ineffective in treating spasms associated with brain or spinal cord diseases or cerebral palsy in children. /US product label does not include/
Tinnitus is defined as an intrinsic sensation of sound that cannot be attributed to an external sound source. There is currently no standardized drug therapy for tinnitus. Based on the similarity between pain and tinnitus, it is recommended that tricyclic antidepressants, as they have analgesic effects, be given particular attention among all antidepressants used to treat tinnitus. This study aims to investigate the effect of a tricyclic antidepressant—cyclobenzapine—on relieving tinnitus symptoms. This study compared 65 patients receiving drug treatment with 30 untreated patients waiting for treatment. Analysis showed that cyclobenzapine was beneficial in reducing both the intensity and distress of tinnitus in the patients, while no improvement was observed in the treatment-waiting control group: 24% of tinnitus patients responded significantly to cyclobenzapine, with a 53% reduction in tinnitus intensity; 25% of patients responded significantly to cyclobenzapine, with a 55% reduction in tinnitus distress. The study also showed that specific subgroups, namely patients with pure-tone tinnitus and those with unilateral tinnitus, responded better to cyclobenzapine. Our results indicate that cyclobenzapine is a promising tinnitus treatment, particularly suitable for certain specific subgroups of patients. Given the favorable risk-benefit ratio of cyclobenzapine and the current lack of a mature and effective treatment for tinnitus, further research is warranted.

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Drug Warning
It has been reported that life-threatening serotonin syndrome may occur when cyclobenzaridine hydrochloride is used in combination with other medications, such as selective serotonin reuptake inhibitors (SSRIs), serotonin-norepinephrine reuptake inhibitors (SNRIs), tricyclic antidepressants (TCAs), tramadol, bupropion, meperidine, verapamil, or monoamine oxidase inhibitors. Concomitant use of cyclobenzaridine hydrochloride with monoamine oxidase inhibitors is contraindicated. Symptoms of serotonin syndrome may include altered mental status (e.g., confusion, agitation, hallucinations), autonomic dysfunction (e.g., excessive sweating, tachycardia, blood pressure fluctuations, hyperthermia), neuromuscular abnormalities (e.g., tremor, ataxia, hyperreflexia, clonic seizures, myotonia), and/or gastrointestinal symptoms (e.g., nausea, vomiting, diarrhea). If any of these reactions occur, cyclobenzaridine hydrochloride and any concomitant serotonergic medications should be discontinued immediately, and symptomatic and supportive treatment should be initiated.
If clinical necessity dictates the concurrent use of cyclobenzafine hydrochloride and other serotonergic drugs, close monitoring is recommended, especially during the initial stages of treatment or dose escalation. In controlled studies, 21% and 32% of patients, respectively, experienced dry mouth after receiving 5 mg and 10 mg cyclobenzafine, compared to 7% in the placebo group. In clinical studies or post-marketing surveillance, 27% and 7% of patients, respectively, experienced dry mouth after receiving 10 mg cyclobenzafine. In controlled studies, 1% to 3% of patients received 5 mg or 10 mg cyclobenzafine and experienced abdominal pain, acid reflux, dyspepsia, constipation, diarrhea, nausea, and dysgeusia; these adverse reactions were also observed in post-marketing surveillance of patients receiving 10 mg cyclobenzafine. Less than 1% of patients receiving 10 mg of this drug during post-marketing surveillance or in controlled studies reported adverse reactions such as vomiting, anorexia, gastrointestinal pain, gastritis, thirst, glossitis, and flatulence. Adverse reactions such as paralytic ileus, tongue discoloration, stomatitis, and parotid gland swelling have been reported in patients receiving other tricyclic antidepressants or, rarely, cyclobenzafine, but their causal relationship with cyclobenzafine could not be established. In patients receiving 10 mg of the drug during postmarketing surveillance or in controlled studies, less than 1% reported adverse reactions including malaise, seizures, ataxia, dizziness, dysarthria, increased muscle tone, tremor, disorientation, insomnia, depressed mood, paresthesia, anxiety, agitation, psychosis, thought disorders, abnormal dreams, hallucinations, excitement, paresthesia, and diplopia. Other adverse neurological reactions have also been reported in some patients receiving other tricyclic antidepressants or, in very rare cases, cyclobenzafine, but their causal relationship with cyclobenzafine could not be established. These reactions include decreased or increased libido, gait abnormalities, delusions, aggressive behavior, paranoia, peripheral neuropathy, Bell's palsy, altered EEG patterns, and extrapyramidal symptoms.
In controlled studies, headache occurred in 5% of patients treated with 5 or 10 mg cyclobenzapine; in controlled studies and post-marketing surveillance, headache occurred in 1-3% of patients treated with 10 mg cyclobenzapine. Adverse reactions such as irritability, decreased mental acuity, nervousness, fatigue, and confusion occurred in 1-3% of patients treated with 5 or 10 mg cyclobenzapine in controlled studies or post-marketing surveillance.
For more complete data on cyclobenzapine warnings (25 in total), please visit the HSDB record page.

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Pharmacodynamics
Cyclobenzapine is a skeletal muscle relaxant that acts on the brainstem to relieve skeletal muscle spasms, but its exact pharmacodynamic mechanism is currently unclear. Although it has a long half-life, its duration of action is relatively short, typically lasting 4-6 hours. Cyclobenzapine has been reported to cause serotonin syndrome when used in combination with other serotonergic drugs. Symptoms of serotonin syndrome may include autonomic instability, altered mental status, neuromuscular abnormalities, or gastrointestinal symptoms—if any of these reactions occur during treatment, cyclobenzapine should be discontinued immediately.

C20H21N

275.39

275.167

303-53-7

303-53-7;6202-23-9 (HCl);

2895

Typically exists as solid at room temperature

1.096 g/cm3

405.9ºC at 760 mmHg

177.8ºC

1.7500 (estimate)

4.553

3.24

0

1

3

21

365

0

Cl.CN(CC/C=C1\C=C2C=CC=CC2=CC2=CC=CC=C\12)C

JURKNVYFZMSNLP-UHFFFAOYSA-N

InChI=1S/C20H21N/c1-21(2)15-7-12-20-18-10-5-3-8-16(18)13-14-17-9-4-6-11-19(17)20/h3-6,8-14H,7,15H2,1-2H3

N,N-dimethyl-3-(2-tricyclo[9.4.0.03,8]pentadeca-1(15),3,5,7,9,11,13-heptaenylidene)propan-1-amine

MK-130MK130Flexeril

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)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

注意: 如下所列的是一些常用的体内动物实验溶解配方，主要用于溶解难溶或不溶于水的产品（水溶度<1 mg/mL）。 建议您先取少量样品进行尝试，如该配方可行，再根据实验需求增加样品量。

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注射用配方1: DMSO : Tween 80： Saline = 10 : 5 : 85 (如： 100 μL DMSO → 50 μL Tween 80 → 850 μL Saline)
*生理盐水/Saline的制备：将0.9g氯化钠/NaCl溶解在100 mL ddH ₂ O中，得到澄清溶液。
注射用配方 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL DMSO → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline)
注射用配方 3: DMSO : Corn oil = 10 : 90 (如： 100 μL DMSO → 900 μL Corn oil)
示例: 以注射用配方 3 (DMSO : Corn oil = 10 : 90) 为例说明, 如果要配制 1 mL 2.5 mg/mL的工作液, 您可以取 100 μL 25 mg/mL 澄清的 DMSO 储备液，加到 900 μL Corn oil/玉米油中, 混合均匀。

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注射用配方 4: DMSO : 20% SBE-β-CD in Saline = 10 : 90 [如：100 μL DMSO → 900 μL (20% SBE-β-CD in Saline)]
*20% SBE-β-CD in Saline的制备（4°C，储存1周）：将2g SBE-β-CD (磺丁基-β-环糊精) 溶解于10mL生理盐水中，得到澄清溶液。
注射用配方 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (如： 500 μL 2-Hydroxypropyl-β-cyclodextrin (羟丙基环胡精)→ 500 μL Saline)
注射用配方 6: DMSO : PEG300 : Castor oil : Saline = 5 : 10 : 20 : 65 (如： 50 μL DMSO → 100 μL PEG300 → 200 μL Castor oil → 650 μL Saline)
注射用配方 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (如： 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
注射用配方 8: 溶解于Cremophor/Ethanol (50 : 50), 然后用生理盐水稀释。
注射用配方 9: EtOH : Corn oil = 10 : 90 (如： 100 μL EtOH → 900 μL Corn oil)
注射用配方 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL EtOH → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline)

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口服配方 1: 悬浮于0.5% CMC Na (羧甲基纤维素钠)
口服配方 2: 悬浮于0.5% Carboxymethyl cellulose (羧甲基纤维素)
示例: 以口服配方 1 (悬浮于 0.5% CMC Na)为例说明, 如果要配制 100 mL 2.5 mg/mL 的工作液, 您可以先取0.5g CMC Na并将其溶解于100mL ddH2O中，得到0.5%CMC-Na澄清溶液；然后将250 mg待测化合物加到100 mL前述 0.5%CMC Na溶液中，得到悬浮液。

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口服配方 3: 溶解于 PEG400 (聚乙二醇400)
口服配方 4: 悬浮于0.2% Carboxymethyl cellulose (羧甲基纤维素)
口服配方 5: 溶解于0.25% Tween 80 and 0.5% Carboxymethyl cellulose (羧甲基纤维素)
口服配方 6: 做成粉末与食物混合

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注意: 以上为较为常见方法，仅供参考， InvivoChem并未独立验证这些配方的准确性。具体溶剂的选择首先应参照文献已报道溶解方法、配方或剂型，对于某些尚未有文献报道溶解方法的化合物，需通过前期实验来确定（建议先取少量样品进行尝试），包括产品的溶解情况、梯度设置、动物的耐受性等。

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。