JAK2 (IC50 = 2.8 nM); JAK1 (IC50 = 3.3 nM); Tyk2 (IC50 = 19 nM); JAK3 (IC50 = 428 nM)

Ruxolitinib 会导致细胞凋亡呈剂量依赖性增加，使 Ba/F3 细胞中线粒体去极化的细胞倍增，并对 JAK2V617F 介导的信号传导和增殖产生强大且特异性的抑制作用。 Ruxolitinib 可减少正常供体和真性红细胞增多症患者的红细胞祖细胞增殖，IC50 值分别为 407 nM 和 223 nM，并表现出显着的抗红细胞集落形成作用，IC50 为 67 nM [1]。

在 JAK2V617F 驱动的小鼠模型中，rufolitinib（180 mg/kg，口服，每日两次）不会引起骨髓抑制或免疫抑制，但它确实通过减少脾肿大和炎症细胞因子的循环水平并优先消除肿瘤细胞来显着延长生存期。第 22 天时，存活率高于 90% [1]。在骨髓纤维化双盲试验中，鲁索替尼组中 41.9% 的患者和安慰剂组中 0.7% 的患者达到了主要终点。 Ruxolitinib 将总体症状评分改善 50% 或更多，同时保持脾脏体积缩小 [2]。

生化测定[1]
用N-末端表位标签通过PCR克隆人JAK1（837-1142）、JAK2（828-1132）、JAK3（781-1124）和Tyk2（873-1187）的激酶结构域。使用Sf21细胞和杆状病毒载体表达重组蛋白，并用亲和层析纯化。JAK激酶测定使用肽底物（-EQUEDEPEGDYFEWLE）的均匀时间分辨荧光测定。用测试化合物或对照、JAK酶、500nM肽、三磷酸腺苷（ATP；1mM）和2.0%二甲基亚砜（DMSO）进行每种酶反应1小时。50%抑制浓度（IC50）计算为抑制50%荧光信号所需的化合物浓度。CHK2和c-MET酶的生化测定使用标准条件（Michaelis常数[Km]ATP）进行，具有来自每种蛋白质和合成肽底物的重组表达的催化结构域
使用标准条件（CEREP；www.CEREP.com）使用200nM INCB018424进行额外的激酶测定（Abl、Akt1、AurA、AurB、CDC2、CDK2、CDK4、CHK2、c-kit、c-Met、EGFR、EphB4、ERK1、ERK2、FLT-1、HER2、IGF1R、IKKα、IKKβ、JAK2、JAK3、JNK1、Lck、MEK1、p38α、p70S6K、PKA、PKCα、Src和ZAP70）。显著抑制被定义为与对照值相比大于或等于30%（重复测定的平均值）。

细胞增殖测定[1]
将细胞接种在2000/孔的白色底部96孔板上，用来自DMSO储备的化合物（0.2%的最终DMSO浓度）处理，并在37°C下用5%CO2孵育48小时。通过使用Cell Titer Glo萤光素酶试剂的细胞ATP测定或活细胞计数来测量存活率。将值转换为相对于车辆控制的抑制百分比，并根据使用PRISM GraphPad的数据的非线性回归分析拟合IC50曲线<小时> 细胞凋亡[1]
膜联蛋白V染色。将细胞处理20至24小时，并用膜联蛋白V和碘化丙啶染色，分别用于分析早期凋亡和死亡细胞。使用FACSCaliber流式细胞仪进行分析。 线粒体膜电位。将细胞处理24小时，然后与2μM染料JC-1一起孵育。使用488nm激发和530nm和585nm发射滤光片通过流式细胞术进行分析。JC-1在线粒体中表现出电位依赖性积累，其发射在红色光谱（590nM）中。从红色（590nM）到绿色（530nM）的荧光转移表明，由于线粒体膜电位的丧失，染料重新分布到细胞质中，线粒体膜电位是细胞凋亡的早期标志物<小时> 菌落形成测定[1]
通过Ficoll离心从PV患者或正常对照者的外周血中分离单核细胞。将来自对照组或PV患者的总共2×105个细胞接种到补充有重组细胞因子（50 ng/mL干细胞因子、10 ng/mL粒细胞-巨噬细胞集落刺激因子、10 mg/mL粒细胞集落刺激因素、10 ng/mL IL-3和3 U/mL红细胞生成素）和指定浓度的INCB018424或DMSO载体的甲醇培养基H88434上。为了评估内源性红系集落生长，将PV患者的3至4×105个细胞接种到含有INCB018424或载体的最小方法培养基上。每种条件一式三份。14天后对来源于红系（突发形成单位[BFU]和集落形成单位[CFU]-E）和髓系（CFU-粒细胞-巨噬细胞）祖细胞的集落进行计数。

JAK2V617F-driven mouse model
In vivo treatment with INCB018424 in a myeloproliferative neoplasm mouse model
All of the procedures were conducted in accordance with the US Public Health Service Policy on Humane Care and Use of Laboratory Animals. Mice were fed standard rodent chow and provided with water ad libitum. Ba/F3-JAK2V617F cells (105 per mouse) were inoculated intravenously into 6- to 8-week-old female BALB/c mice. Survival was monitored daily, and moribund mice were humanely killed and considered deceased at time of death. Treatment with vehicle (5% dimethyl acetamide, 0.5% methocellulose) or INCB018424 began within 24 hours of cell inoculation, twice daily by oral gavage. Hematologic parameters were measured using a Bayer Advia120 analyzed, and statistical significance was determined using Dunnett testing[1].

Absorption, Distribution and Excretion
Following oral administration, ruxolitinib undergoes rapid absorption and the peak concentrations are reached within one hour after administration. Over a single-dose range of 5 mg to 200 mg, the mean maximal plasma concentration (Cmax) increases proportionally. Cmax ranged from 205 nM to 7100 nM and AUC ranged from 862 nM x hr to 30700 nM x hr. Tmax ranges from one to two hours following oral administration. Oral bioavailability is at least 95%.
Following oral administration of a single radiolabeled dose of ruxolitinib, the drug was mainly eliminated through metabolism. About 74% of the total dose was excreted in urine and 22% was excreted in feces, mostly in the form of hydroxyl and oxo metabolites of ruxolitinib. The unchanged parent drug accounted for less than 1% of the excreted total radioactivity.
The mean volume of distribution (%coefficient of variation) at steady-state is 72 L (29%) in patients with myelofibrosis and 75 L (23%) in patients with polycythemia vera. It is not known whether ruxolitinib crosses the blood-brain barrier.
Ruxolitinib clearance (% coefficient of variation) is 17.7 L/h in women and 22.1 L/h in men with myelofibrosis. Drug clearance was 12.7 L/h (42%) in patients with polycythemia vera and 11.9 L/h (43%) in patients with acute graft-versus-host disease.
Following oral administration, absorption of ruxolitinib is approximately 95%, and mean systemic bioavailability is estimated to be about 80%. Following oral administration of ruxolitinib, peak plasma concentrations are achieved within 1-2 hours. ... Following administration of a single oral dose of radiolabeled ruxolitinib in healthy individuals, elimination was predominantly through metabolism with 74 and 22% of radioactivity excreted in urine and feces, respectively. Unchanged drug accounted for less than 1% of the excreted total radioactivity.

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Metabolism / Metabolites
More than 99% of orally-administered ruxolitinib undergoes metabolism mediated by CYP3A4 and, to a lesser extent, CYP2C9. The major circulating metabolites in human plasma were M18 formed by 2-hydroxylation, and M16 and M27 (stereoisomers) formed by 3-hydroxylation. Other identified metabolites include M9 and M49, which are formed by hydroxylation and ketone formation. Not all metabolite structures are fully characterized and it is speculated that many metabolites exist in stereoisomers. Metabolites of ruxolitinib retain inhibitory activity against JAK1 and JAk2 to a lesser degree than the parent drug.
Cytochrome P-450 (CYP) isoenzyme 3A4 is the major enzyme responsible for metabolism of ruxolitinib. Two major active metabolites were identified in the plasma of healthy individuals; all active metabolites contribute 18% of the overall pharmacodynamic activity of ruxolitinib.
Ruxolitinib is metabolized mainly by cytochrome P-450 (CYP) isoenzyme 3A4.

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Biological Half-Life
The mean elimination half-life of ruxolitinib is approximately 3 hours and the mean half-life of its metabolites is approximately 5.8 hours.
The mean half-life of ruxolitinib following a single oral dose is approximately 3 hours, and the mean half-life of ruxolitinib and its metabolites is approximately 5.8 hours.

Toxicity Summary
IDENTIFICATION AND USE: Ruxolitinib phosphate is used for the treatment of intermediate- or high-risk myelofibrosis, including primary myelofibrosis, post-polycythemia vera myelofibrosis, and post-essential thrombocythemia myelofibrosis. Ruxolitinib is designated an orphan drug by the US Food and Drug Administration (FDA) for use in the treatment of these conditions. HUMAN EXPOSURE AND TOXICITY: Adverse effects reported in more than 10% of patients receiving ruxolitinib include thrombocytopenia, anemia, neutropenia, bruising, dizziness, and headache. Ruxolitinib-treated patients achieved clinically meaningful improvements in myelofibrosis-related symptoms and quality of life, but patients receiving placebo reported worsening of symptoms and other patient-reported outcomes. Ruxolitinib, as compared with placebo, provided significant clinical benefits in patients with myelofibrosis by reducing spleen size, ameliorating debilitating myelofibrosis-related symptoms, and improving overall survival. These benefits came at the cost of more frequent anemia and thrombocytopenia in the early part of the treatment period. In vitro data indicate that neither ruxolitinib nor its M18 metabolite is an inhibitor of P-glycoprotein (P-gp), breast cancer resistance protein (BCRP), organic anion-transporting polypeptides (OATP) 1B1 and 1B3, organic cation transporters (OCT) 1 and 2, and organic anion transporters (OAT) 1 and 3 at clinically relevant concentrations. Ruxolitinib also is not a substrate for P-gp. It was not clastogenic in in vitro chromosomal aberration assay (cultured human peripheral blood lymphocytes). ANIMAL STUDIES: Ruxolitinib was not mutagenic in a bacterial mutagenicity assay (Ames test) or clastogenic in vivo in a rat bone marrow micronucleus assay. Ruxolitinib was not carcinogenic in the 6-month Tg.rasH2 transgenic mouse model study or in a 2-year carcinogenicity study in the rat. In a pre- and post-natal development study in rats, pregnant animals were dosed with ruxolitinib from implantation through lactation at doses up to 30 mg/kg/day. There were no drug-related adverse findings in pups for fertility indices or for maternal or embryofetal survival, growth and development parameters at the highest dose evaluated (34% the clinical exposure at the maximum recommended dose of 25 mg twice daily). Ruxolitinib was administered orally to pregnant rats or rabbits during the period of organogenesis, at doses of 15, 30 or 60 mg/kg/day in rats and 10, 30 or 60 mg/kg/day in rabbits. There was no evidence of teratogenicity. However, decreases of approximately 9% in fetal weights were noted in rats at the highest and maternally toxic dose of 60 mg/kg/day. This dose results in an exposure (AUC) that is approximately 2 times the clinical exposure at the maximum recommended dose of 25 mg twice daily. In rabbits, lower fetal weights of approximately 8% and increased late resorptions were also noted at the highest and maternally toxic dose of 60 mg/kg/day.

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Hepatotoxicity
In the large clinical trials, serum ALT elevations occurred in 25% to 48% of ruxolitinib treated subjects versus 7% to 9% of placebo recipients. The ALT elevations were generally self-limited, asymptomatic and mild and were above 5 times ULN in only 1.3% of patients. In the prelicensure clinical trials, no cases of clinically apparent liver injury were reported. Among causes of death in one trial of ruxolitinib for myelofibrosis, one was attributed to hepatic failure; this instance, however, was considered unrelated to ruxolitinib. Since its approval and more wide scale use, rare cases of clinically apparent ruxolitinib induced acute liver injury have been reported, but without documentation of clinical features or careful exclusion of other causes. Importantly, there also have been several published reports of reactivation of hepatitis B, in patients with and without HBsAg (but with anti-HBc) in serum. A rise in HBV DNA levels was identified within 1 to 6 months of starting ruxolitinib and was associated with elevations in ALT levels and jaundice in some patients. HBV DNA levels decreased rapidly upon starting anti-HBV therapy with entecavir and all patients recovered. In one instance, HBV DNA levels declined with lowering of the dose of ruxolitinib, but then rose again when the dose was increased.
So far, reports of the use of ruxolitinib for COVID-19 have included only small numbers of patients and have provided little information on hepatic adverse events or risk of reactivation of hepatitis B.
Likelihood score: C (probable cause of reactivation of hepatitis B in susceptible patients).

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Effects During Pregnancy and Lactation
◉ Summary of Use during Lactation
No information is available on the clinical use of ruxolitinib during breastfeeding. Because ruxolitinib is 97% bound to plasma proteins, the amount in milk is likely to be low. The manufacturer recommends that breastfeeding be discontinued during ruxolitinib therapy and for 2 weeks after the last dose for the oral tablets and for 4 weeks after the last dose for the topical cream.
◉ 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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Protein Binding
Ruxolitinib is approximately 97% bound to plasma proteins, mostly to albumin.

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Interactions
Fluconazole: The AUC of ruxolitinib is predicted to increase by approximately 100% to 300% following concomitant administration with the combined CYP3A4 and CYP2C9 inhibitor fluconazole at doses of 100 mg to 400 mg once daily, respectively. Avoid the concomitant use of Jakafi with fluconazole doses of greater than 200 mg daily.
Concomitant administration of ruxolitinib (single 50-mg dose) with rifampin (600 mg once daily for 10 days) decreased ruxolitinib peak plasma concentrations and AUC by 32 and 61%, respectively. No dosage adjustment is recommended when ruxolitinib is administered with a CYP3A4 inducer.
Concomitant administration of ruxolitinib (single 10-mg dose) with erythromycin (500 mg twice daily for 4 days) increased ruxolitinib peak plasma concentrations and AUC by 8 and 27%, respectively. No dosage adjustment is recommended when ruxolitinib is administered with weak or moderate CYP3A4 inhibitors (e.g., erythromycin). In patients receiving a stable dosage of ruxolitinib, clinicians should use caution when initiating treatment with a moderate CYP3A4 inhibitor, especially in patients with low platelet counts.
Concomitant administration of ruxolitinib (single 10-mg dose) with ketoconazole (200 mg twice daily for 4 days) increased peak plasma concentrations and AUC of ruxolitinib by 33 and 91%, respectively. Half-life of ruxolitinib also was prolonged from 3.7 to 6 hours with concomitant use of ketoconazole. Dosage reduction is recommended when ruxolitinib is administered with potent CYP3A4 inhibitors (e.g., ketoconazole).
Concomitant use of ruxolitinib with potent inhibitors of CYP3A4 (e.g., boceprevir, clarithromycin, conivaptan, grapefruit juice, indinavir, itraconazole, ketoconazole, lopinavir/ritonavir, mibefradil [no longer commercially available in the US], nefazodone, nelfinavir, posaconazole, ritonavir, saquinavir, telaprevir, telithromycin, voriconazole) has resulted in increased peak plasma concentrations and area under the serum concentration-time curve (AUC) of ruxolitinib. Dosage reduction of ruxolitinib is recommended when the drug is used concomitantly with potent CYP3A4 inhibitors.

[1]. Quintas-Cardama A, et al. Preclinical characterization of the selective JAK1/2 inhibitor INCB018424: therapeutic implications for the treatment of myeloproliferative neoplasms. Blood, 2010, 115(15), 3109-3117.
[2]. Verstovsek S, et al. A double-blind, placebo-controlled trial of ruxolitinib for myelofibrosis. N Engl J Med, 2012, 366(9), 799-807.
[3]. Tavallai M, et al. Rationally Repurposing Ruxolitinib (Jakafi (®)) as a Solid Tumor Therapeutic.Front Oncol. 2016 Jun 13;6:14

Therapeutic Uses
/CLINICAL TRIALS/ ClinicalTrials.gov is a registry and results database of publicly and privately supported clinical studies of human participants conducted around the world. The Web site is maintained by the National Library of Medicine (NLM) and the National Institutes of Health(NIH). Each ClinicalTrials.gov record presents summary information about a study protocol and includes the following: Disease or condition; Intervention (for example, the medical product, behavior, or procedure being studied); Title, description, and design of the study; Requirements for participation (eligibility criteria); Locations where the study is being conducted; Contact information for the study locations; and Links to relevant information on other health Web sites, such as NLM's MedlinePlus for patient health information and PubMed for citations and abstracts for scholarly articles in the field of medicine. Ruxolitinib is included in the database.
Jakafi is indicated for treatment of patients with polycythemia vera who have had an inadequate response to or are intolerant of hydroxyurea. /Included in US product label/
Jakafi is indicated for treatment of patients with intermediate or high-risk myelofibrosis, including primary myelofibrosis, post-polycythemia vera myelofibrosis and post-essential thrombocythemia myelofibrosis. /Included in US product label/
Ruxolitinib phosphate is used for the treatment of intermediate- or high-risk myelofibrosis, including primary myelofibrosis, post-polycythemia vera myelofibrosis, and post-essential thrombocythemia myelofibrosis. Ruxolitinib is designated an orphan drug by the US Food and Drug Administration (FDA) for use in the treatment of these conditions.
Ruxolitinib, a Janus kinase (JAK) 1 and 2 inhibitor, was shown to have a clinical benefit in patients with polycythemia vera in a phase 2 study. We conducted a phase 3 open-label study to evaluate the efficacy and safety of ruxolitinib versus standard therapy in patients with polycythemia vera who had an inadequate response to or had unacceptable side effects from hydroxyurea. We randomly assigned phlebotomy-dependent patients with splenomegaly, in a 1:1 ratio, to receive ruxolitinib (110 patients) or standard therapy (112 patients). The primary end point was both hematocrit control through week 32 and at least a 35% reduction in spleen volume at week 32, as assessed by means of imaging. The primary end point was achieved in 21% of the patients in the ruxolitinib group versus 1% of those in the standard-therapy group (P<0.001). Hematocrit control was achieved in 60% of patients receiving ruxolitinib and 20% of those receiving standard therapy; 38% and 1% of patients in the two groups, respectively, had at least a 35% reduction in spleen volume. A complete hematologic remission was achieved in 24% of patients in the ruxolitinib group and 9% of those in the standard-therapy group (P=0.003); 49% versus 5% had at least a 50% reduction in the total symptom score at week 32. In the ruxolitinib group, grade 3 or 4 anemia occurred in 2% of patients, and grade 3 or 4 thrombocytopenia occurred in 5%; the corresponding percentages in the standard-therapy group were 0% and 4%. Herpes zoster infection was reported in 6% of patients in the ruxolitinib group and 0% of those in the standard-therapy group (grade 1 or 2 in all cases). Thromboembolic events occurred in one patient receiving ruxolitinib and in six patients receiving standard therapy. In patients who had an inadequate response to or had unacceptable side effects from hydroxyurea, ruxolitinib was superior to standard therapy in controlling the hematocrit, reducing the spleen volume, and improving symptoms associated with polycythemia vera.

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Drug Warnings
Ruxolitinib can cause adverse hematologic reactions, including thrombocytopenia, anemia, and neutropenia. A complete blood cell count (CBC) must be performed before initiating therapy with ruxolitinib.
Patients should be assessed for the risk of developing serious bacterial, mycobacterial, fungal, and viral infections. Active serious infections should have resolved prior to initiating therapy with ruxolitinib. Clinicians should carefully observe patients receiving ruxolitinib for signs and symptoms of infection and should promptly initiate appropriate treatment.
Herpes zoster infection occurred in 1.9% of patients receiving ruxolitinib in a clinical study. Clinicians should inform patients about the early signs and symptoms of herpes zoster and advise patients to seek treatment as soon as possible for this condition.
Following interruption or discontinuance of ruxolitinib therapy, symptoms of myelofibrosis generally return to pretreatment levels within approximately 1 week. Withdrawal manifestations, characterized by acute relapse of disease symptoms, accelerated splenomegaly, worsening of cytopenias, and occasional hemodynamic decompensation (including septic shock-like syndrome with severe hypoxia, hypotension, fever, and confusion), have been reported in some patients following discontinuance of ruxolitinib. Some experts recommend that ruxolitinib dosage should be tapered gradually over a 2-week period under close medical supervision.
For more Drug Warnings (Complete) data for RUXOLITINIB (9 total), please visit the HSDB record page.

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Pharmacodynamics
Ruxolitinib is an antineoplastic agent that inhibits cell proliferation, induces apoptosis of malignant cells, and reduces pro-inflammatory cytokine plasma levels by inhibiting JAK-induced phosphorylation of signal transducer and activator of transcription (STAT). Inhibition of STAT3 phosphorylation, which is used as a marker of JAK activity, by ruxolitinib is achieved at two hours after dosing which returned to near baseline by 10 hours in patients with myelofibrosis and polycythemia vera. In clinical trials, ruxolitinib reduced splenomegaly and improved symptoms of myelofibrosis. In a mouse model of myeloproliferative neoplasms, administration of ruxolitinib was associated with prolonged survival. Ruxolitinib inhibits both mutant and wild-type JAK2; however, JAK2V617F mutation, which is often seen in approximately 50% of patients with myelofibrosis, was shown to reduce ruxolitinib sensitivity, which may also be associated with possible resistance to JAK inhibitor treatment.

C17H18N6

306.3650

306.159

C, 66.65; H, 5.92; N, 27.43

941678-49-5

Ruxolitinib (S enantiomer);941685-37-6;Ruxolitinib phosphate;1092939-17-7;(Rac)-Ruxolitinib-d9;2469553-67-9;Deuruxolitinib-d8;1513883-39-0;Ruxolitinib sulfate;1092939-16-6

25126798

Colorless oil

1.4±0.1 g/cm3

592.6±50.0 °C at 760 mmHg

312.2±30.1 °C

0.0±1.7 mmHg at 25°C

1.747

1.69

83.18

1

4

4

23

453

1

[C@@H](C1CCCC1)(N1N=CC(C2N=CN=C3NC=CC=23)=C1)CC#N

HFNKQEVNSGCOJV-OAHLLOKOSA-N

InChI=1S/C17H18N6/c18-7-5-15(12-3-1-2-4-12)23-10-13(9-22-23)16-14-6-8-19-17(14)21-11-20-16/h6,8-12,15H,1-5H2,(H,19,20,21)/t15-/m1/s1

(R)-3-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-3-cyclopentylpropanenitrile

INCB-018424, INCB 018424, INCB 18424, INCB-18424; INCB018424; INC424, INC424, INC-424; INCB18424, Jakafi and Jakavi (trade name)

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: 61 mg/mL (199.1 mM) Water:<1 mg/mL Ethanol:<1 mg/mL

配方 1 中的溶解度: 5 mg/mL (16.32 mM) in 5% DMAC in 0.5% methylcellulose aqueous solution (这些助溶剂从左到右依次添加，逐一添加), 悬浮液；超声助溶。

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配方 2 中的溶解度: ≥ 2.08 mg/mL (6.79 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 20.8 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 3 中的溶解度: ≥ 2.08 mg/mL (6.79 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 20.8 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 4 中的溶解度: ≥ 2.08 mg/mL (6.79 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 20.8 mg/mL 澄清 DMSO 储备液加入900 μL 玉米油中，混合均匀。

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配方 5 中的溶解度: 2% DMSO+30% PEG 300+ddH2O:5mg/mL

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配方 6 中的溶解度: 5 mg/mL (16.32 mM) in 0.5% Methylcellulose/saline water (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.
*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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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。