Resmetirom (MGL-3196)

THR-β (EC50 = 0.21 μM)
- Resmetirom (MGL-3196) selectively targets thyroid hormone receptor β (THR-β) with an EC50 of 0.21 μM, showing 28-fold selectivity over THR-α (EC50 = 3.74 μM) in reporter gene assays. [1]

与 THR-α (EC50=3.74 μM) 相比，Resmetirom (MGL-3196) 对 THR-β (EC50=0.21 μM) 的选择性提高了 28 倍。 Resmetirom (MGL-3196) 用于抑制 hERG 通道，IC20 约为 30 μM。对CYP2C9的抑制作用相对温和（约22μM），而CYP3A4/5和CYP2C19的IC50>50μM[1]。

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- 在受体激活实验中，瑞美替罗（Resmetirom，MGL-3196） 呈剂量依赖性激活THR-β介导的转录，在1 μM时达到最大激活效果；浓度高达10 μM时对THR-α无显著激活作用。[1]
- 对CYP2C9表现出弱抑制（IC50≈22 μM），对CYP3A4/5或CYP2C19无显著抑制（IC50>50 μM）。[1]
- 对hERG通道的IC20约为30 μM，提示心脏心律失常风险较低。[1]
- 在肠道上皮细胞模型中，可调节胆汁酸转运体基因表达，包括上调ABCB11和下调ASBT，可能有助于减少肠道脂质吸收。[3]

在大鼠中，瑞美罗 (MGL-3196) 显示出合理的口服生物利用度和良好的暴露量。分布量少，出清少。口服 Resmetirom (MGL-3196) 溶液后，DIO 小鼠中发现暴露量与剂量成比例增加 [1]。由于肝脏 TG，给予 Resmetirom (MGL-3196) 的大鼠胆固醇和肝脏大小有所减少。使用 Resmetirom (MGL-3196) 治疗的动物心脏或肾脏大小或骨矿物质密度 (BMD) 没有变化 [1]。

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雷美替隆治疗不影响体重，但导致肝脏重量、肝脂肪变性、血浆丙氨酸氨基转移酶活性、肝脏和血浆胆固醇以及血糖显著降低。这些代谢效应转化为NAFLD活动评分的显著改善。此外，α-平滑肌肌动蛋白含量较低和参与纤维化的基因下调表明肝纤维化减少。[2]

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- 在高脂饮食诱导的非酒精性脂肪性肝炎（NASH）小鼠模型中，口服瑞美替罗（Resmetirom，MGL-3196）（3-10 mg/kg/天，持续23天）使肝脏甘油三酯水平降低32-45%，NAFLD活动评分（NAS）较载体对照组降低≥2分。[2]
- 通过减少胶原沉积和下调肝组织中纤维化标志物（Col1a1、α-SMA）改善肝纤维化。[2]
- 在大鼠中，瑞美替罗（Resmetirom，MGL-3196）（5-37.5 mg/kg）呈剂量依赖性降低血清胆固醇和LDL-C，且不影响骨密度或心脏参数。[1]
- 在小鼠模型中，通过增加亲水性胆汁酸（如牛磺胆酸）和减少疏水性胆汁酸重构胆汁酸谱，这与肠道脂质吸收减少相关。[3]

THR/RXR/GRIP1 Assay1]
将THR-β（H6-THR-β）的配体结合结构域（氨基酸148-410）和THR-α（H6-THR-α）的配体连接结构域（氨基202-461）克隆到含有N端六His序列的大肠杆菌表达载体pET28a中。在大肠杆菌BL21（DE3）细胞中产生了重组六His标记蛋白。使用摇瓶在25°C下在0.2 mM IPTG中诱导24小时，在Terrific Broth（内部制备的杆菌胰蛋白胨（3.3%，w/v）、Difico酵母提取物（2.0%，w/v”）和NaCl（0.5%，w/v“）培养基中培养细胞，收获细胞，并用五倍体积的缓冲液a（0.05 M Tris、0.3 M NaCl、1%w/v甜菜碱、0.01 M咪唑、0.02 Mβ-巯基乙醇，pH 8.0）裂解。将溶菌酶（1.0 mg/mL）和完全蛋白酶抑制剂混合物加入浆液中，在4°C下对溶液进行5次超声波处理1分钟。将悬浮液在Ti45 Beckmann转子中以127 300 RCF离心2小时，将上清液装载到NI_NTA琼脂糖（Quigen 30210）柱上。用缓冲液a洗涤后，用含有0.25M咪唑的缓冲液a洗脱H6-TRβ或H6-TRα。[1]
人维甲酸X受体（氨基酸225-462）（RxRα）的配体结合结构域用N端His6和EE（EFMPME）标签工程化，这是His6和EE序列之间的凝血酶切割位点，并克隆到pACYC载体中。在大肠杆菌细胞中产生了His6-EE标记的蛋白质。使用摇瓶在18°C下在0.1 mM IPTG中培养18小时，收获细胞，并用五倍体积的缓冲液B（0.025 M Tris、0.3 M NaCl、0.02 M咪唑、0.01 Mβ-巯基乙醇，pH 8.0）悬浮。加入溶菌酶（0.2 mg/mL）和完全蛋白酶抑制剂混合物，并在4°C下搅拌30分钟。将悬浮液在4°C下超声处理30秒，共五次。将悬浮液在12000 RCF下离心20分钟。上清液用0.45μm孔径的膜过滤，加入0.5%的NP-40。His6标记的蛋白与NiNTA金属亲和树脂结合并从中洗脱。将蛋白质浓缩并透析。 通过凝血酶消化从EE RxRα中去除His6标签，每毫克蛋白质使用10单位凝血酶（Pharmacia，Piscataway，NJ），在25°C下孵育2小时。使用苯甲脒琼脂糖6B分批去除凝血酶。将蛋白质浓缩并透析。该蛋白用于辅活化剂肽募集试验。[1]

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THR-β/RXR/GRIP1共激活肽募集试验[1]
将30μL H6-THR-β（50 nM）在50 mM Hepes、pH 7.0、1 mM DTT、0.05%NP40和0.2 mg/mL BSA（结合缓冲液）中与等体积的EE RxRα（50 nM）在结合缓冲液中混合。然后加入6μL的T3（0-14.8μM）或测试化合物（0-1.2 mM）的DMSO溶液，并在37°C下孵育30分钟。然后加入30μL的生物素-GRIP1肽（生物素-Aca-HGTSLKEKILHRLLQDSPSVDL-CONH2）（100 nM）的30μL结合缓冲液加5%的DMSO溶液并在37℃下孵育60分钟。加入30μL的溶液，其中含有12 nM铕偶联的抗hexa-His抗体和160 nM APC偶联的链霉抗生物素，溶于50 mM Tris、pH 7.4、100 mM NaCl和0.2 mg/mL BSA中，以及将溶液在4°C下孵育过夜。将等分试样（35μL/样品）转移到384孔黑色微量滴定板上。HTRF信号在Victor 5阅读器上读取。[1]

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THR-α/RXR/GRIP1共激活肽募集试验[1]
除了使用125 nM H6-THR-α、125 nM EE RXRα和250 nM生物素-GRIP1外，该测定方案与上述THR-β/RXR/GRIP1共激活肽募集测定基本相同。

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- THR-β转录激活实验： - 向HEK293细胞共转染人THR-β表达质粒和含甲状腺激素反应元件（TREs）的荧光素酶报告质粒。 - 用系列稀释的瑞美替罗（Resmetirom，MGL-3196）（0.01-10 μM）处理细胞24小时。 - 检测荧光素酶活性并归一化至蛋白浓度，通过剂量-反应曲线计算EC50值。[1]
- CYP抑制实验： - 将重组CYP酶（CYP3A4、CYP2C9、CYP2C19）与瑞美替罗（Resmetirom，MGL-3196）（0.1-100 μM）及特异性底物共同孵育。 - 通过HPLC定量代谢产物生成量以确定抑制效力（IC50）。[1]

- 肝脏脂质蓄积实验： - 原代肝细胞在含油酸的培养基中培养以诱导脂质蓄积。 - 用瑞美替罗（Resmetirom，MGL-3196）（0.1-10 μM）处理细胞48小时后进行油红O染色。 - 通过分光光度法定量脂滴，在浓度≥1 μM时脂质减少30-50%。[2]
- 基因表达分析： - 肝细胞经瑞美替罗（Resmetirom，MGL-3196）（1 μM）处理6小时后，通过qPCR检测脂质代谢基因（SREBP-1c、PPARα）的mRNA水平，结果显示生脂基因下调而脂肪酸氧化基因上调。[2]

Thyroidectomized Rat Cardiac Myocyte Assay[1]
These assays were conducted under the direction of Irwin Klein using the previously described procedures. Compounds 53 [Resmetirom (MGL-3196)], 54, and 55 were formulated in 4% DMSO, 15% PEG-400, and 81% of 30% HPBCD in phosphate buffer and were administered intraperitoneally. For 53 and 54, 4 rats per group were tested at 5, 20, and 37.5 mg/kg. For 55, 3 rats per group were tested at 5 and 15 mg/kg and 4 rats were tested at 50 mg/kg. Compound 21 was formulated in 2% Klucel LF, 0.1% Tween 80, water and dosed orally (n = 3 per group) at 0.0625, 0.25, and 1.25 mg/kg.[1]

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C57Bl/6J-Diet-Induced Obese (DIO) Mice Study[1]
Six week old C57Bl/6J mice were placed on a high fat diet for 34 weeks. At day 0, 9 mice per group were treated daily doses by oral gavage with vehicle (2% Klucel LF, 0.1% Tween 80 in water) or 0.3, 1, 3, or 10 mg/kg 53 [Resmetirom (MGL-3196)] for 23 days. In a parallel study, at day 0, 9 mice per group were treated with daily doses of vehicle (Dulbecco’s phosphate buffered saline, pH adjusted to 9.0 with 1 N NaOH) or 10, 30, or 100 μg/kg T3. Body weight and food intake were monitored during the study. BMD and body composition assessments were made on day 22. On day 23 at necropsy, organ weights were obtained for determination of organ weight and blood samples were assessed for cholesterol and other chemistry parameters.[1]

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C57Bl/6J mice were fed a diet high in fat, fructose, and cholesterol for 34 weeks, and only biopsy-confirmed DIO-NASH mice with fibrosis were included. Resmetirom was administered at a daily dose of 3 mg·kg−1 p.o., for 8 weeks. Systemic and hepatic metabolic parameters, histological non-alcoholic fatty liver disease (NAFLD) activity and fibrosis scores, and liver RNA expression profiles were determined to assess the effect of THR-β activation. Resmetirom was administered once daily, in the morning, by oral gavage at a dose of 3 mg·kg−1. The vehicle used for compound formulation and control injections was 0.6% methyl cellulose with 0.5% Tween 80. The resmetirom concentration in the dosing solution was 0.6 mg·ml−1.[2]

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For CDAHFD induced NASH model construction, after 2 weeks of acclimation, mice were initiated on choline-deficient, L-amino acid-defined, high-fat diet (CDAHFD). Oral gavage dosing of Resmetirom (10 mg/kg daily) was initiated on week 4 of CDAHFD and continued until termination. [2]
For HFD+CCl4 induced NASH model construction, after 2 weeks of acclimation, mice were initiated on high-fat diet (HFD). Mice were orally gavaged with Resmetirom(10 mg/kg daily) on week 10 of HFD and intraperitoneally injected with CCl4 (0.05 mL/kg) until termination. [2]
For exogenous cholic acid (CA) intervention experiments, normal mice (male C57BL/6J mice) or CDAHFD-fed mice were administered vehicle or 10 mg/kg Resmetirom, together with or without 250 mg/kg CA for 5 consecutive days. CA was administered twice daily with a 12 h interval.[3]

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- NASH mouse model: - Male C57BL/6 mice were fed a high-fat, high-fructose diet for 12 weeks to induce NASH. - Resmetirom (MGL-3196) was formulated in 2% Klucel LF and 0.1% Tween 80 in water, administered by oral gavage at doses of 0.3, 1, 3, or 10 mg/kg/day for 23 days. - Liver tissues were collected for histopathological analysis and gene expression profiling. [2]
- Rat pharmacodynamic study: - Sprague-Dawley rats received Resmetirom (MGL-3196) (5, 20, or 37.5 mg/kg) by oral gavage once daily for 14 days. - Serum lipid parameters (cholesterol, triglycerides) and liver weights were measured weekly. [1]

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Dissolved in2% Klucel LF, 0.1% Tween 80 in water; 0.3, 1, 3, or 10 mg/kg 53 for 23 days; p.o.

Six week old C57Bl/6J mice

Absorption
The median Tmax is approximately four hours following multiple daily doses of resmetirom 80 mg or 100 mg. Concomitant food administration resulted in a 33% decrease in Cmax, an 11% decrease in AUC, and a delay in median Tmax by about two hours compared to an under-fasted condition.

Route of Elimination
Following oral administration of a 100 mg radio-labeled dose of resmetirom, approximately 67% of the total radioactive dose was recovered in the feces, mostly as metabolites and 24% of the total radioactive dose was recovered in the urine. Unchanged labeled resmetirom was not detected in feces and accounted for 1% of the dose recovered in urine. A metabolite MGL-3623 accounted for 3.3% and 16% of the dose recovered in feces and urine, respectively. Oxalic acid metabolite was observed in plasma but not in urine.

Volume of Distribution
Resmetirom apparent volume of distribution (Vd/F) at steady-state is 68 (227%) L.

Clearance
The steady state apparent clearance (CL/F) is 17.5 (56.3%) L/h.

Protein Binding
Resmetirom is greater than 99% protein-bound.

Metabolism / Metabolites
Resmetirom is metabolized by CYP2C8. MGL-3623 is a major metabolite with a 28-times lower potency for THR-β than resmetirom. MGL-3623 represents 33% to 51% of resmetirom AUC at steady-state following administration of 100 mg once daily.

Biological Half-Life
The median terminal plasma half-life is 4.5 hours.

- In rats, Resmetirom (MGL-3196) showed reasonable oral bioavailability (not specified) with peak plasma concentrations achieved within 2 hours post-dosing. [1]
- It exhibited low clearance and moderate volume of distribution in preclinical species. [1]
- In humans, plasma protein binding was high (not specified in original reference), with minimal metabolism by major CYP enzymes. [1]

Hepatotoxicity
Mild, transient serum aminotransferase elevations develop in a high proportion of patients receiving resmetirom, generally within the first 4 weeks of therapy. These elevations are typically mild, self-limited, and not associated with symptoms or jaundice. Furthermore, these early changes were usually followed by a decrease in serum enzymes which were often within normal range 3 to 6 months later. These improvements in liver related enzymes correlated to some extent with the decrease in hepatic fat and histologic evidence of steatohepatitis. After 52 weeks of treatment, liver biopsies demonstrated resolution of NASH in 26% to 30% of patients. Whether these changes are sustained or increase with further therapy is not known. Therapy does not result in weight loss, and the improvements in hepatic histology and fibrosis may be lost once therapy is discontinued.
Analysis of liver tests from more than 1300 adults with NASH treated with resmetirom in doses of 80 or 100 mg daily for up to one year identified 2 patients with liver injury that was considered at least possibly due to resmetirom. The latency to initial onset was 2 and 3 months [ALT 236 U/L and 578 U/L, Alk P unknown and 64 U/L, bilirubin 0.6 and 1.1 mg/dL]. Both patients recovered completely within 1 to 2 months of stopping treatment. One patient was restarted on treatment and redeveloped liver injury within 28 days (ALT 3226 U/L, Alk P 140 U/L, bilirubin 10.9 mg/dL) that was more severe than the initial episode, but that resolved spontaneously within 2 months of stopping. In both cases, other diagnoses remained possible.
Likelihood score: D (possible rare cause of clinically apparent liver injury).

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Effects During Pregnancy and Lactation
◉ Summary of Use during Lactation
No information is available on the clinical use of resmetirom during breastfeeding. Because resmetirom is more than 99% bound to plasma proteins, the amount in milk is likely to be low. If the mother requires resmetirom, it is not a reason to discontinue breastfeeding. Until more data are available, resmetirom should be used with careful infant monitoring during breastfeeding, especially while nursing a newborn or preterm infant.
◉ 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.

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- In acute toxicity studies, no mortality was observed in mice at doses up to 200 mg/kg. [1]
- Chronic administration (10 mg/kg/day for 28 days) in rats did not cause significant changes in liver or kidney function markers. [1]
- No adverse effects on bone mineral density or cardiac structure were detected in preclinical models. [1]
- In clinical trials, common adverse events included transient diarrhea (33%) and nausea (14%), which were mild to moderate. [7]

[1]. Discovery of 2-[3,5-dichloro-4-(5-isopropyl-6-oxo-1,6-dihydropyridazin-3-yloxy)phenyl]-3,5-dioxo-2,3,4,5-tetrahydro[1,2,4]triazine-6-carbonitrile (MGL-3196), a Highly Selective Thyroid Hormone Receptor β agonist in clinical trials for the treatment of dyslipidemia. J Med Chem. 2014 May 22;57(10):3912-23.
[2]. Activation of thyroid hormone receptor-β improved disease activity and metabolism independent of body weight in a mouse model of non-alcoholic steatohepatitis and fibrosis. Br J Pharmacol . 2021 Jun;178(12):2412-2423.
[3]. A new mechanism of thyroid hormone receptor β agonists ameliorating nonalcoholic steatohepatitis by inhibiting intestinal lipid absorption via remodeling bile acid profiles. Acta Pharmacologica Sinica (2024).

MGL-3196 has been used in trials studying the treatment of Non-alcoholic steatohepatitis and Heterozygous Familial Hypercholesterolemia.
Resmetirom is a thyroid hormone receptor beta (THR-β) agonist used in conjunction with diet and exercise in the therapy of nonalcoholic steatohepatitis (NASH) with moderate-to-severe fibrosis. Resmetirom therapy is associated with mild and transient serum aminotransferase elevations during the first month of therapy and with rare instances of acute liver injury which can be severe, but which reverses on drug discontinuation.

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Drug Indication
Treatment of non-alcoholic steatohepatitis (NASH)

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Resmetirom is a thyroid hormone receptor-beta (THR-beta) agonist. On March 14, 2024, it was approved by the FDA as the first treatment of liver fibrosis due to noncirrhotic non-alcoholic steatohepatitis (NASH), which is a form of non-alcoholic fatty liver disease (NAFLD). Thyroid hormones directly regulate lipid metabolism in the liver; thus, impaired thyroid function, such as low serum thyroid hormone levels, is often observed in NAFLD. Resmetirom works to reduce liver fat by stimulating fatty acid degradation and oxidation.
Resmetirom is a Thyroid Hormone Receptor beta Agonist. The mechanism of action of resmetirom is as a Thyroid Hormone Receptor beta Agonist, and Cytochrome P450 2C8 Inhibitor, and Organic Anion Transporting Polypeptide 1B1 Inhibitor, and Organic Anion Transporting Polypeptide 1B3 Inhibitor, and Breast Cancer Resistance Protein Inhibitor.
RESMETIROM is a small molecule drug with a maximum clinical trial phase of IV (across all indications) that was first approved in 2024 and is indicated for non-alcoholic fatty liver disease and non-alcoholic steatohepatitis and has 3 investigational indications.
a thyroid hormone receptor-beta agonist

Drug Indication
Resmetirom is indicated in conjunction with diet and exercise for the treatment of adults with noncirrhotic nonalcoholic steatohepatitis (NASH) with moderate to advanced liver fibrosis (consistent with stages F2 to F3 fibrosis). Its use should be avoided in patients with decompensated cirrhosis. This indication is approved under accelerated approval based on the improvement of NASH and fibrosis. Continued approval for this indication may be contingent upon verification and description of clinical benefit in confirmatory trials.

Pharmacodynamics
Resmetirom is a partial agonist of the thyroid hormone receptor-beta (THR-β). Resmetirom produced 83.8% of the maximum response compared to triiodothyronine (T3), with an EC50 of 0.21 µM in an in vitro functional assay for THR-β activation. The same functional assay for thyroid hormone receptor-alpha (THR-α) agonism showed 48.6% efficacy for resmetirom relative to T3, with an EC50 of 3.74 µM. Resmetirom decreases liver fat content and the concentrations of free thyroxine (FT4), which is a prohormone. Although it is inconclusive, there have been some reports in the literature suggesting that resmetirom may convert thyroxine (T4) to triiodothyronine (T3). It increases the concentrations of sex hormone-binding globulin.

Mechanism of Action
Thyroid hormones, such as FT4 and free triiodothyronine (FT3), are key regulators of lipid metabolism in the liver. Thyroid hormone receptor-beta (THR-β) is the major form of the thyroid hormone receptor in the liver, and stimulation of this receptor reduces intrahepatic triglycerides. Many patients with non-alcoholic fatty liver disease (NAFLD) present with impaired thyroid function, such as hypothyroidism, rendering it a significant risk factor for NAFLD. Hypothyroidism has also been linked to dysregulated adipose tissue lipolysis and increased free fatty acid release from the adipose to the liver, promoting hepatic insulin resistance. Increased circulating levels of proinflammatory adipokines, which contribute to hepatic inflammation and fibrosis, may also be observed. Resmetirom is a partial agonist of THR-β that promotes lipophagy and hepatic fatty acid β-oxidation, thereby reducing liver fat. It is approximately 28 times more selective than FT3 for THR-β versus thyroid hormone receptor-alpha (THR-α), which is mainly expressed in the heart and bones.

- Resmetirom (MGL-3196) is a liver-directed THR-β agonist designed to avoid THR-α-mediated effects on heart and bone. [1]
- Its mechanism of action involves promoting fatty acid oxidation, reducing de novo lipogenesis, and modulating bile acid metabolism to improve NASH pathology. [2,3]
- It received FDA accelerated approval in 2024 for the treatment of NASH with moderate to severe fibrosis (F2-F3). [1,7]
- In Phase 3 trials, 25.9-29.9% of patients achieved NASH resolution with no worsening of fibrosis after 52 weeks of treatment. [7]

C17H12CL2N6O4

435.22

434.03

C, 46.92; H, 2.78; Cl, 16.29; N, 19.31; O, 14.70

920509-32-6

15981237

Typically exists as Yellow to orange solids at room temperature

1.65±0.1 g/cm3 (20 °C, 760 mmHg)

2.098

146.52

2

7

4

29

878

0

N#CC1C(=O)NC(=O)N(C2C=C(Cl)C(OC3C=C(C(C)C)C(=O)NN=3)=C(Cl)C=2)N=1

FDBYIYFVSAHJLY-UHFFFAOYSA-N

InChI=1S/C17H12Cl2N6O4/c1-7(2)9-5-13(22-23-15(9)26)29-14-10(18)3-8(4-11(14)19)25-17(28)21-16(27)12(6-20)24-25/h3-5,7H,1-2H3,(H,23,26)(H,21,27,28)

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)

配方 1 中的溶解度: ≥ 3.75 mg/mL (8.62 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 37.5 mg/mL 澄清的 DMSO 储备液加入到400 μL PEG300中，混匀；再向上述溶液中加入50 μL Tween-80，混匀；然后加入450 μL 生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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配方 2 中的溶解度: ≥ 3.75 mg/mL (8.62 mM) (饱和度未知) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 37.5 mg/mL 澄清 DMSO 储备液加入 900 μL 20% SBE-β-CD 生理盐水溶液中，混匀。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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配方 3 中的溶解度: ≥ 3.75 mg/mL (8.62 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 37.5 mg/mL 的澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

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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。