CA IX/carbonic anhydrase (IC50 = 30 nM)

乙酰唑胺还抑制 hCA II，IC50 为 130 nM[1]。乙酰唑酰胺（Ace）作为一种微小的杂芳族磺酰胺，与多种碳酸酐酶具有很强的亲和力结合，并抑制碳酸酐酶（CA）的活性[2]。与对照组相比，高乙酰唑酰胺浓度（AceH，50 nM）、顺铂（Cis；1 μg/mL）以及 Cis 与低乙酰唑酰胺浓度（AceL，10 nM）联合处理显着降低了 Hep-2 细胞的活力。对照组[2]。用乙酰唑胺/Cis 组合治疗后，P53 表达水平显着升高，如与对照组相比，AceL+Cis 和 AceH+Cis 治疗后 P53 蛋白表达水平显着升高所见。与对照组相比，Ace/Cis 组合治疗显着降低了 bcl-2/bax 表达比率，并增强了 caspase-3 蛋白的表达。与对照组相比，AceL、AceH、Cis 和 AceL+Cis 疗法显着降低了 bcl-2/bax 比率[2]。 Ace 和 Cis 联合治疗可有效促进 Hep-2 细胞凋亡[2]。 Ace/Cis 联合治疗显着降低 Hep-2 细胞中 AQP1 mRNA 的表达。在 Hep-2 细胞中，与对照组相比，AceH 和 AceL+Cis 处理均降低了水通道蛋白-1 (AQP1) mRNA 的表达[2]。

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Acetazolamide (AZ) /乙酰唑胺， MS-275和AZ + MS-275处理抑制NB SH-SY5Y细胞的生长。 AZ、MS-275和AZ + MS-275处理降低NB SH-SY5Y细胞的迁移能力。 结果：我们评估了HDAC抑制剂（HDACi） pyridylmethyl-N-{4-[（2-氨基苯基）-氨基甲酰]-benzyl}-氨基甲酸酯（MS-275）与pan CA抑制剂乙酰唑酰胺（acetazolamide， AZ）联合对NB SH-SY5Y、SK-N-SH和SK-N-BE(2)细胞的抗肿瘤潜力。关键观察结果是AZ + MS-275联合用药显著抑制NB细胞株SH-SY5Y的生长，诱导细胞周期阻滞和凋亡，降低迁移能力。[2]

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本研究的目的是确定Acetazolamide (AZ) /乙酰唑胺（Ace）治疗是否增强Hep-2喉部细胞对顺铂（Cis）的化疗敏感性。在对数生长期，Hep-2细胞分别用Ace、Cis或两者处理，用MTT法检测细胞活力。流式细胞术检测细胞凋亡程度。western blotting检测凋亡相关蛋白BCL2凋亡调节因子（bcl-2）、BCL2相关X （bax）和caspase-3的表达水平，增殖相关蛋白增殖细胞核抗原（PCNA）和肿瘤蛋白p53 （p53）的表达水平。采用逆转录-聚合酶链反应检测各组水通道蛋白-1 (AQP1) mRNA表达水平。与单独使用药物相比，Ace和Cis联合治疗对Hep-2细胞的生长抑制和诱导凋亡具有协同作用。Ace/Cis联合应用可降低PCNA的表达，增加p53的表达。此外，联合处理降低了bcl-2/bax的比值，增加了caspase-3的表达，降低了AQP1的表达。这些结果表明，Ace和Cis联合使用可增强喉癌细胞的化疗敏感性。[3]

在神经母细胞瘤 (NB) SH-SY5Y 异种移植物中，乙酰唑胺 (40 mg/kg) 大大增强了 MS-275 对肿瘤发生的抑制作用 [3]。 ？在 NB SH-SY5Y 异种移植物中，乙酰唑胺 (40 mg/kg) 和/或 MS-275 治疗可降低 HIF1-α 和 CAIX 表达 [3]。 ？在 NB SH-SY5Y 异种移植物中，乙酰唑胺 (40 mg/kg)、MS-275 和乙酰唑胺+MS-275 会降低有丝分裂和增殖标记物的表达[3]。 ？当给予乙酰唑胺（50 mg/kg；口服）三天时，受感染小鼠阴道中的淋球菌负荷显着减少 90%[6]。

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淋球菌感染是世界范围内一项紧迫的公共卫生威胁，因为感染发病率不断增加，同时细菌对大多数抗生素的耐药性也在增加。这导致有效的治疗选择越来越少。毫无疑问，迫切需要开发新的、有效的抗淋球菌药物。为了发现新的抗淋球菌治疗药物，我们先前鉴定了乙酰唑胺，一种碳酸酐酶抑制剂，作为一种新的淋病奈瑟菌抑制剂。乙酰唑胺在0.5 ~ 4 μg/mL浓度范围内抑制淋病奈瑟菌的生长，表现出较强的体外抗淋球菌活性。本研究的目的是研究乙酰唑胺对淋病奈瑟菌生殖道感染小鼠模型的体内疗效。与用药物治疗的小鼠相比，乙酰唑胺在治疗三天后显著减少了感染小鼠阴道内90%的淋球菌负荷。这些结果表明，乙酰唑胺作为一种有希望的治疗选择值得进一步研究，以补充有限的抗淋球菌治疗药物。[6]

AlamarBlue细胞毒性试验[2]
标准方案按描述执行。用AlamarBlue试剂观察Acetazolamide (AZ) /乙酰唑胺（Acetazolamide， AZ）、MS-275和AZ + MS-275处理的细胞与对照（DMSO- 0.2x10−4μ m）的存活率，每孔中加入占总体积10%的试剂4 h后进行荧光检测。采用SPECTRAmax Gemini分光光度计(激发波长540 nm；发射波长590nm)。

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碘化丙啶细胞周期测定[2]
简单地说，用Acetazolamide (AZ)和/或MS-275处理过的2 × 106细胞用柠檬酸盐盐水提振，在80%的低温乙醇中固定48小时，然后将细胞制成颗粒，在2mg /mL的RNase A中重新悬浮5分钟。加入0.1 mg/mL的碘化丙啶溶液，在RT下孵育30分钟，细胞通过细胞过滤器过滤到5ml的聚苯乙烯管中。标记的细胞在BD FACSCAN流式细胞仪上分析。数据采用FlowJo软件上的Watson-Pragmatic模型进行拟合。

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创面愈合试验[2]
将SH-SY5Y细胞接种于48孔板上的玻璃片上，以105个细胞/孔的密度在500 μl培养基中贴敷过夜，一式三份。井的底部用一条黑色直线作了定位。在90%的融合时，用200 μl的移液管尖端用标记向导划伤细胞单层。用培养基洗去松散的非贴壁细胞。在培养基中加入新鲜培养基，添加乙酰唑胺(AZ) (10 μM, 20 μM, 40 μM)和MS-275 (0.75 μM， 1.5 μM和3 μM)，培养48 h。48 h后，用PBS洗涤细胞，4%多聚甲醛固定。在PBS中洗涤三次后，细胞用20%甲醇中1%结晶紫染色。在治疗0、48和72 h的时间点拍摄10倍原始放大倍数的相对比光显微图像。使用NIH Image J程序对迁移的细胞进行人工计数，以量化迁移到伤口区域的细胞数量。每个实验进行了三次，一式三份，并显示了一个代表性的分析。

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对于药物处理，Hep-2细胞用Acetazolamide (ACE)(低浓度1×10−8 mol/l，这里称为Acetazolamide (ACE) l；或高浓度5×10−8 mol/l，这里称为AceH), Cis（1µg/ml）单独，或Cis与Ace （AceL+Cis，或AceH+Cis）联合作用48小时。用等体积的媒介物处理的细胞作为对照。所有实验中Ace浓度分别为1×10−8或5×10−8 mol/l。所有实验均以1µg/ml的浓度使用Cis。Cis和Ace均溶解于二甲亚砜（DMSO）中，然后加入PBS稀释至最终工作浓度。培养物中DMSO的终浓度不超过0.5%。用AceH单独、Cis单独或联合（AceH+Cis）或对照（对照）治疗HUVECs 48 h。[3]

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Annexin V凋亡测定[3]
凋亡细胞采用Annexin V-异硫氰酸荧光素(FITC)/碘化丙啶（PI）双染色，采用FITC-Annexin V凋亡检测试剂盒定量。对数生长期，将Hep-2细胞置于6孔板中。细胞分别用Acetazolamide (ACE)L、AceH、Cis、AceL+Cis、AceH+Cis或对照液处理48 h。然后用PBS洗涤细胞，用胰蛋白酶消化，用富钙HEPES缓冲液重悬。按照制造商的说明，用Annexin V-FITC和PI染色该悬液15分钟。最后用FlowJo软件对细胞进行分析。

Xenograft studies for determining the in vivo efficacy of Acetazolamide (AZ) , MS-275, and Acetazolamide (AZ)  + MS-275 combination [2]
For the in vivo xenograft study, 4–6 weeks-old female NOD/SCID mice were obtained from the animal facility. Subcutaneous xenograft tumors were developed by injecting SH-SY5Y cells (2 × 106) into the inguinal fat pad of NOD/SCID mice. When tumor diameter reached 0.5 cm, the mice were randomized into four groups (5 mice per group). The control and treatment groups received intraperitoneal injections of vehicle (PBS) or Acetazolamide (AZ) (40 mg/kg), MS-275 (20 mg/kg) or the combination, respectively, every day for 2 weeks. Experiments were terminated when tumor sizes exceeded 2 cm3 in volume or animals showed signs of morbidity. Tumor diameters were measured on a daily basis until termination. The long (D) and short diameters (d) were measured with calipers. Tumor volume (cm3) was calculated as V = 0.5 × D × d2. After euthanizing the mice, tumors were resected, weighed and fixed in 10% neutral-buffered formalin at room temperature and processed for histopathology. For the in vivo serial heterotransplantation analysis, 2x106 untreated and pretreated Acetazolamide (AZ)  + MS-275 cells, manually and enzymatically dissociated from treated tumors, were injected subcutaneously to NOD/SCID mice. Growth rates were measured 2–3 times per week. On the 38th day, the animals were sacrificed, after which tumors were removed and weighed.

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Mice infection and treatment [6]
Two days after pellet implantation (Day 0), the vagina of each mouse was rinsed with 50 mM HEPES (pH = 7.4), and each mouse was inoculated intravaginally with 20 μL of the prepared bacterial suspension of N. gonorrhoeae FA1090 (1.2 × 108 CFU/mL). Two days post-infection (Day +2), mice were randomly allocated into groups (n=10) and administered either Acetazolamide (50 mg/kg) or the vehicle (DMSO-Tween 80-PBS, 1:1:8) orally for three consecutive days. As a positive control, one group of mice was administered a single intraperitoneal dose of ceftriaxone (15 mg/kg in water).

Absorption, Distribution and Excretion
Carbonic anhydrase inhibitors are avidly bound by carbonic anhydrase and, accordingly, tissues rich in this enzyme will have higher concentrations of carbonic anhydrase inhibitors following systemic administration. /Carbonic Anhydrase Inhibitors/
Inhibitors of Carbonic Anhydrase. Drug: acetazolamide; Oral Absorption: nearly complete; Plasma Half-Life: 6-9 hours; and Route of Elimination: renal excretion of intact drug. /From table/
ACETAZOLAMIDE RELATED TO RESPONSE IN RABBIT; KIDNEY RESPONSE, MEASURED BY MONITORING URINE FLOW & NA ELIMINATION, URINE FLOW & NA ELIMINATION OCCUR IMMEDIATELY AFTER INJECTION CORRELATED WITH LOG DOSE.
IV BOLUS INJECTIONS OF (14)C-LABELED, ACETAZOLAMIDE WERE MADE IN RABBITS. PLASMA, URINE, & WASHED RED BLOOD CELL CONCN WERE MEASURED, THE LATTER INDICATING BOUND DRUG.
For more Absorption, Distribution and Excretion (Complete) data for ACETAZOLAMIDE (6 total), please visit the HSDB record page.

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Metabolism / Metabolites
ACETAZOLAMIDE DOSE NOT UNDERGO METABOLIC ALTERATION.

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Biological Half-Life
3 to 9 hours
Plasma half-life: 6-9 hours /From table/

Toxicity Summary
IDENTIFICATION: Acetazolamide is of synthetic origin. Acetazolamide exists as a white to faintly yellowish-white, odorless crystalline powder. Acetazolamide is very slightly soluble in water and only slightly soluble in ethanol (approx 750 g/l); it is practically insoluble in ether and chloroform. Uses: Preoperative management of closed-angle glaucoma, or as an adjunct in the treatment of open-angle glaucoma. Abnormal retention of fluid: drug-induced edema, obesity, and congestive cardiac failure. Epilepsy Metabolic alkalemia. Periodic paralysis HUMAN EXPOSURE: Patients with either acute or chronic overdosage with acetazolamide may show signs of dehydration with thirst, lethargy, confusion, poor skin turgor, and prolonged capillary refill time, but may have a paradoxical continued diuresis. Electrolyte abnormalities include hyponatremia, hypokalemia, and a non-anion gap hyperchloremic metabolic acidosis in the more than mild ingestion which may lead to further deterioration in mental status, production of seizures, electrocardiographic abnormalities, and arrhythmias. Prior renal insufficiency will lead to increased toxicity at a given dose. There are idiosyncratic reactions producing bone marrow suppression with hepatic and renal insufficiency. Acetazolamide may also precipitate in the renal tubules producing calculi with renal colic. Hypokalemia may lead to muscular weakness, hyporeflexia, and hypochloremic metabolic alkalosis. In geriatric patients, a chronic metabolic acidosis may lead to a chronic compensatory hyperventilation which increases pulmonary vascular resistance and decreases left ventricular function. This can be especially significant in patients on concurrent beta-blocker or calcium channel blocker therapy. The ventricular fibrillation threshold may then be reduced. Cardiac arrhythmias may occur due to potassium deficiency. Abuse or overdose may result in pancreatitis. Hyperglycemia, hyperuricemia, and hyperlipidemia may occur with acute overdose or in chronic use or abuse. Hypersensitivity reactions such as rash, photosensitivity, thrombocytopenia, and pancreatitis are rare. Contraindications: Renal hyperchloremic acidosis. Addison's disease and all types of suprarenal gland failure. Conditions where there is known depletion of sodium and potassium (at least until this is treated). Long-term administration is contraindicated in patients with chronic closed angle-closure glaucoma. Known sensitivity to sulfonamides. Acetazolamide should not be used to alkalinize urine following salicylate overdose since it may worsen metabolic acidosis. Acetazolamide is well absorbed from the gastrointestinal tract. Acetazolamide is distributed throughout body tissues; it concentrates principally in erythrocytes, plasma and kidneys and to a lesser extent in liver, muscles, eyes and the central nervous system. Acetazolamide does not accumulate in tissues. The drug crosses the placenta in unknown quantities. Acetazolamide is tightly bound to carbonic anhydrase and high concentrations are present in tissues containing this enzyme such as erythrocytes and the renal cortex. There is a small amount of irreversible binding to red cells. It is 70 to 90% bound to plasma protein. Acetazolamide is not metabolized. Acetazolamide is excreted unchanged by the kidneys via tubular secretion and passive reabsorption. There is no evidence of enterohepatic circulation although small amounts of unchanged drug are eliminated in the bile. Acetazolamide is a carbonic anhydrase inhibitor. Acetazolamide reduces the formation of hydrogen and bicarbonate ions from carbon dioxide and water by noncompetitive, reversible inhibition of the enzyme carbonic anhydrase, thereby reducing the availability of these ions for active transport into secretions. One patient died of cholestatic jaundice after taking 13 g of acetazolamide in 26 days. In one patient, fatal bone marrow depression with leukopenia, thrombocytopenia, and anemia occurred after therapy with 500 mg of acetazolamide twice daily for 14 weeks. One case of renal failure (anuria) occurred in a patient after taking 500 mg of acetazolamide twice daily for 2 weeks. There have been no reports of congenital defects despite past widespread use though one women on 750 mg per day for glaucoma during the 1st and 2nd trimester had a baby with a sacrococcygeal teratoma but no causal link could be made. Teratogenicity tests in rats and mice showed the absence of fourth and fifth digits from the right forelimb in the offspring of rats and mice. There were no apparent lessions in the newborn of rabbits and monkeys. The drug crosses the placenta in unknown quantities. Potentially hazardous interactions: The effects of folic acid antagonists, oral hypoglycaemic agents and oral anticoagulants may be increased by acetazolamide. The urinary antiseptic effect of methenamine may be prevented by acetazolamide by keeping the urine alkaline. The alkalinization of the urine by acetazolamide can reduce the urinary excretion of many weak bases (including amphetamine, quinine, quinidine, and diethylcarbamazine) and thus enhance their pharmacological effects. In one patient taking phenytoin and acetazolamide drug-induced osteomalacia was reported. The more serious effects include blood disorders, skin toxicity and renal stone formation. Stevens-Johnson syndrome has not been reported. Symptomatic adverse effects: Flushing, thirst, headache, drowsiness, dizziness, fatigue, irritability, excitement, paresthesias, ataxia, hyperpnoa and gastrointestinal disturbances have all been reported (Dollery). Oral ingestion is the usual means of exposure. There is no appreciable dermal absorption. There is no significant absorption or local irritation. ANIMAL/PLANT STUDIES: Numerous animal studies have demonstrated that the toxicity of acetazolamide was very low in the species studied (mouse, dog, rat, monkey).

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Interactions
/ACETAZOLAMIDE/ POTENTIATES MERCURIAL DIURETICS /IN ANIMALS/.
Concurrent use /of glucocorticoid corticosteroids, especially with significant mineralocorticoid activity; mineralocorticoid corticosteroids; parenteral amphotericin; corticotropin, especially prolonged therapeutic use/ with carbonic anhydrase inhibitors may result in severe hypokalemia and should be undertaken with caution; serum potassium concentrations and cardiac function should be monitored during concurrent use. /Carbonic anhydrase inhibitors/
Concurrent use of corticosteroids or corticotropin with acetazolamide sodium may increase the risk of hypernatremia and/or edema because these medications cause sodium and fluid retention; the risk with corticosteroids or corticotropin may depend on the patients's sodium requirement as determined by the condition being treated. /Acetazolamide sodium/
The possibility should be considered that concurrent chronic use of corticosteroids or corticotropin with carbonic anhydrase inhibitors may increase the risk of hypocalcemia and osteoporosis because these medications increase calcium excretion. /Carbonic anhydrase inhibitors/
For more Interactions (Complete) data for ACETAZOLAMIDE (16 total), please visit the HSDB record page.

[1]. Dual-tail approach to discovery of novel carbonic anhydrase IX inhibitors by simultaneously matching the hydrophobic and hydrophilic halves of the active site. Eur J Med Chem. 2017 May 26;132:1-10.

[2]. Acetazolamide potentiates the anti-tumor potential of HDACi, MS-275, in neuroblastoma. BMC Cancer. 2017 Feb 24;17(1):156.

[3]. Combined treatment with acetazolamide and cisplatin enhances chemosensitivity in laryngeal carcinoma Hep-2 cells. Oncol Lett. 2018 Jun;15(6):9299-9306.

[4]. Acetazolamide: a forgotten diuretic agent. Cardiol Rev. 2011 Nov-Dec;19(6):276-8.

[5]. Interaction of antihypertensive acetazolamide with nonsteroidal anti-inflammatory drugs. J Photochem Photobiol B. 2013 Aug 5;125:155-63.

[6]. In vivo efficacy of acetazolamide in a mouse model of Neisseria gonorrhoeae infection. Microb Pathog. 2022 Mar;164:105454.

Therapeutic Uses
Anticonvulsants; Carbonic Anhydrase Inhibitors; Diuretics
MEDICATION (VET): IN LAMINITIS, UDDER EDEMA, ENTEROTOXEMIA, ASCITES, & GLAUCOMA IN VARIOUS SPECIES.
Carbonic anhydrase inhibitors are indicated primarily as adjuncts to other agents in the treatment of open-angle (chronic simple) glaucoma and secondary glaucoma, and to lower intraocular pressure prior to surgery for some types of glaucoma. /Carbonic anhydrase inhibitors; Included in US product labeling/
Acetazolamide is used to lower intraocular pressure in the treatment of malignant (ciliary block) glaucoma, which may occur after inflammation surgery, trauma, or use of miotics. /NOT included in US product labeling/
For more Therapeutic Uses (Complete) data for ACETAZOLAMIDE (13 total), please visit the HSDB record page.

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Drug Warnings
VET: CONTRAINDICATED IN ADRENAL FAILURE OR LOW POTASSIUM AND SODIUM SYNDROMES.
SAFE USE OF THESE AGENTS DURING PREGNANCY HAS NOT BEEN ESTABLISHED. THESE AGENTS ARE CONTRAINDICATED IN PT WITH IDIOPATHIC RENAL HYPERCHLOREMIC ACIDOSIS, RENAL FAILURE, KNOWN DEPLETION OF SODIUM & OF POTASSIUM, ADDISON'S DISEASE, & PT KNOWN TO BE SENSITIVE TO THIS CLASS OF DRUGS. /CARBONIC ANHYDRASE INHIBITORS/
DIURETICS, SUCH AS ACETAZOLAMIDE & THIAZIDES, CAN ALKALINIZE URINE & THUS THEORETICALLY WOULD LIMIT USEFULNESS OF METHENAMINE AS WELL AS ITS MANDELATE & HIPPURATE SALTS AS URINARY TRACT ANTI-INFECTIVE AGENTS.
Maternal Medication usually Compatible with Breast-Feeding: Acetazolamide: Reported Sign or Symptom in Infant or Effect on Lactation: None. /from Table 6/
For more Drug Warnings (Complete) data for ACETAZOLAMIDE (13 total), please visit the HSDB record page.

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Pharmacodynamics
Acetazolamide is a potent carbonic anhydrase inhibitor, effective in the control of fluid secretion, in the treatment of certain convulsive disorders and in the promotion of diuresis in instances of abnormal fluid retention. Acetazolamide is not a mercurial diuretic. Rather, it is a nonbacteriostatic sulfonamide possessing a chemical structure and pharmacological activity distinctly different from the bacteriostatic sulfonamides.

C4H6N4O3S2

222.237

221.988

C, 21.62; H, 2.72; N, 25.21; O, 21.60; S, 28.85

59-66-5

Acetazolamide;59-66-5; 1424-27-7 (sodium)

1986

White to off-white solid powder

1.7±0.1 g/cm3

256-261°C

1.641

-0.26

151.66

2

7

2

13

297

0

BZKPWHYZMXOIDC-UHFFFAOYSA-N

InChI=1S/C4H6N4O3S2/c1-2(9)6-3-7-8-4(12-3)13(5,10)11/h1H3,(H2,5,10,11)(H,6,7,9)

N-(5-sulfamoyl-1,3,4-thiadiazol-2-yl)acetamide

Acetazolamide; Diluran; Diamox; Defiltran; 59-66-5; Diamox; Acetamox; Nephramide; Glaupax; N-(5-Sulfamoyl-1,3,4-thiadiazol-2-yl)acetamide; Acetazolamid; PIM005; Glaupax

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 : ~50 mg/mL (~224.97 mM)
H2O : < 0.1 mg/mL

配方 1 中的溶解度: ≥ 2.5 mg/mL (11.25 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 25.0 mg/mL澄清DMSO储备液加入到400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；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: ≥ 2.5 mg/mL (11.25 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 25.0 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: ≥ 2.5 mg/mL (11.25 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 25.0 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

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配方 4 中的溶解度: 1.96 mg/mL (8.82 mM) in PBS (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液; 超声助溶 (<60°C).

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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。