Indirubin
(Synonyms: 靛玉红; Couroupitine B; Indigo red; Indigopurpurin) 目录号 : GC12975
Indirubin是强效的周期蛋白依赖性激酶（CDKs）（IC50：50-100nmol/L）和糖原合成酶激酶-3β（GSK-3β）（IC50：5-50nmol/L）抑制剂，具有口服给药特性。
Cas No.:479-41-4
Sample solution is provided at 25 µL, 10mM.
;)
_1-7.$$$37316672@70.jpg');)
;)
;)
_273-287.$$$36806353@46.jpg');)
_1-16.$$$37156877@44.jpg');)
;)
;)
;)
_1124-1138.$$$35908550@30.jpg');)
_766-780.$$$37100057@30.jpg');)
_418-432.$$$36893736@30.jpg');)
_240-255.$$$35108512@29.jpg');)
_533-545.$$$36543525@28.jpg');)
_264-276.$$$36600053@22.jpg');)
_2214998.$$$@0.jpg');)
Indirubin is a potent inhibitor of cyclin-dependent kinases (CDKs) (IC50: 50-100nmol/L) and glycogen synthase kinase-3beta (GSK-3β) (IC50: 5-50nmol/L), with oral administration characteristics[1]. Indirubin serves as the primary active ingredient within the traditional Chinese medicine Indigo naturalis and belongs to the bisindole alkaloid category[2]. Indirubin shows promise as a medical treatment against tumors along with inflammatory conditions and neuropathic disorders and is effective against bacterial infections[3].
In vitro, Indirubin showed an IC50 of 12.5μmol/L in glioma U87 and U118 cells for cell proliferation inhibition, and the IC50 in normal astrocytes was 100μmol/L[2]. Indirubin (10μmol/L) activated AhR signaling and inhibited tumor cell proliferation and invasion in clear renal cell carcinoma and osteoma for 24h[4]. Indirubin binds to ATP sites through competitive interactions and suppresses the functions of CDK1/cyclin B, CDK2/cyclin A and CDK2/cyclin E in starfish oocytes at 100μmol/L after 30h treatment[5]. Indirubin increased opsonized zymosan-induced superoxide production in human myelocytic leukaemia HL-60 cells at low concentrations ranging from 10nmol/L to 1μmol/L[6]. Indirubin (1μmol/L, 37 °C for 6 days) suppresses CDK2 expression and Rb protein phosphorylation in HL-60 myeloid leukemia cells while triggering interleukin-8 (IL-8) activity to promote PU.1 activation and neutrophil differentiation[6]. Indirubin (1μmol/L) suppressed KBM-5 human chronic myeloid leukemia cell proliferation by reducing expression levels of NF-κB-dependent protein cyclin D1[6]. Indirubin (1μmol/L, 24h) significantly inhibited the phosphorylation of STAT3 in ovarian cancer cells, and cell apoptosis occurred after treatment of 1μmol/L Indirubin for 72h[7].
In vivo, Indirubin treatment (25-100μmol/L) inhibited the expression of IL-1β, IL-6, and TNF-α in the lipopolysaccharide (LPS)-stimulated mastitis mouse models in a dose-dependent manner[8]. Indirubin（100μmol/L）ameliorated the pathological changes induced by LPS and reduced neutrophils and macrophages in the alveolar space, resulting in a reduction in the thickness of the mammary bleb[8]. Inhibition of intersegmental blood vessel (ISV) formation in zebrafish embryos by treatment with indirubin at 100μmol/L for 24h[9]. Indirubin (100μmol/L) showed mild toxicity in zebrafish embryos[9]. Oral Indirubin (5-10mg/kg, 24h) significantly reduced the anxiety behavior of the male Swiss albino mice and exerted a sedative effect[10].
References:
[1] Leclerc S, Garnier M, Hoessel R, et al. Indirubins Inhibit Glycogen Synthase Kinase-3β and CDK5/P25, Two Protein Kinases Involved in Abnormal Tau Phosphorylation in Alzheimer's Disease: A PROPERTY COMMON TO MOST CYCLIN-DEPENDENT KINASE INHIBITORS?* 210[J]. Journal of Biological Chemistry, 2001, 276(1): 251-260.
[2]Yang L, Li X, Huang W, et al. Pharmacological properties of indirubin and its derivatives[J]. Biomedicine & Pharmacotherapy, 2022, 151: 113112.
[3] Ju Z, Sun J, Liu Y. Molecular structures and spectral properties of natural indigo and indirubin: Experimental and DFT studies[J]. Molecules, 2019, 24(21): 3831.
[4] Ishida M, Mikami S, Shinojima T, et al. Activation of aryl hydrocarbon receptor promotes invasion of clear cell renal cell carcinoma and is associated with poor prognosis and cigarette smoke[J]. International journal of cancer, 2015, 137(2): 299-310.
[5] Hoessel R, Leclerc S, Endicott J A, et al. Indirubin, the active constituent of a Chinese antileukaemia medicine, inhibits cyclin-dependent kinases[J]. Nature cell biology, 1999, 1(1): 60-67.
[6] Suzuki K, Adachi R, Hirayama A, et al. Indirubin, a Chinese anti‐leukaemia drug, promotes neutrophilic differentiation of human myelocytic leukaemia HL‐60 cells[J]. British journal of haematology, 2005, 130(5): 681-690.
[7] Chen L, Wang J, Wu J, et al. Indirubin suppresses ovarian cancer cell viabilities through the STAT3 signaling pathway[J]. Drug design, development and therapy, 2018: 3335-3342.
[8] Lai J, Liu Y, Liu C, et al. Indirubin inhibits LPS-induced inflammation via TLR4 abrogation mediated by the NF-kB and MAPK signaling pathways[J]. Inflammation, 2017, 40: 1-12.
[9] Alex D, Lam I K, Lin Z X, et al. Indirubin shows anti-angiogenic activity in an in vivo zebrafish model and an in vitro HUVEC model[J]. Journal of ethnopharmacology, 2010, 131(2): 242-247.
[10] Disha I J, Hasan R, Bhuia S, et al. Anxiolytic efficacy of Indirubin: in vivo approach along with receptor binding profiling and molecular interaction with GABAergic pathways[J]. ChemistryOpen, 2025, 14(2): e202400290.
Indirubin是强效的周期蛋白依赖性激酶（CDKs）（IC50：50-100nmol/L）和糖原合成酶激酶-3β（GSK-3β）（IC50：5-50nmol/L）抑制剂，具有口服给药特性[1]。Indirubin是中药青黛的主要活性成分，属于双吲哚类生物碱[2]。Indirubin在抗肿瘤、抗炎症及神经病变治疗领域展现出良好前景，同时具有抗细菌感染功效[3]。
在体外，Indirubin对胶质瘤细胞U87和U118的增殖抑制IC50为12.5μmol/L，而在正常星形胶质细胞中IC50达100μmol/L[2]。在透明细胞肾癌和骨瘤模型中，10μmol/L浓度的Indirubin作用24小时可激活AhR信号通路，同时抑制肿瘤细胞增殖与侵袭[4]。100μmol/L浓度的Indirubin处理海星卵母细胞30h后可通过竞争性结合ATP位点，有效抑制CDK1/cyclin B、CDK2/cyclin A 和CDK2/cyclin E的生物学功能[5]。在10nmol/L至1μmol/L的低浓度范围内，Indirubin可增强人髓系白血病HL-60细胞对酵母聚糖诱导的超氧化物生成[6]。Indirubin（1μmol/L，37℃，处理6天）抑制HL-60髓系白血病细胞CDK2表达和Rb蛋白磷酸化，同时激活白细胞介素-8 （IL-8）活性，以促进PU.1活化和中性粒细胞分化[6]。Indirubin（1μmol/L）能通过降低NF-κB依赖性蛋白Cyclin D1的表达，抑制KBM-5人慢性髓系白血病细胞增殖[6]。Indirubin（1μmol/L, 24h）显著抑制卵巢癌细胞STAT3的磷酸化，1μmol/L浓度的Indirubin处理卵巢癌细胞72h后，细胞发生凋亡[7]。
在体内，Indirubin（25-100μmol/L）能够以剂量依赖的方式抑制脂多糖（LPS）诱导的乳腺炎小鼠模型中 IL-1β、IL-6 和 TNF-α 的表达[8]。在100μmol/L浓度下，Indirubin可改善LPS引起的病理损伤，减少肺泡腔中的中性粒细胞和巨噬细胞浸润，降低乳腺泡壁厚度[8]。使用100μmol/L浓度的Indirubin处理斑马鱼胚胎24小时，可抑制斑马鱼胚胎的节间血管（ISV）的形成[9]，100μmol/L浓度的Indirubin对胚胎表现出轻度毒性[9]。口服Indirubin（5-10mg/kg, 24h）能显著缓解瑞士白化雄性小鼠的焦虑行为，发挥镇静作用[10]。
| Cell experiment [1]: | |
Cell lines | HL-60 cells |
Preparation Method | Over a period of 6 days, HL-60 cells transformed into neutrophil-like cells under the influence of 25% DMSO and 25ng/ml G-CSF while being treated with Indirubin (1μmol/L). The generation of Oinline images from differentiated cells stimulated by opsonized zymosan occurred through the reduction process of cytochrome c as detailed in previous research. Fluo-3 fluorescence changes provided measurements for the calcium response of HL-60 cells differentiated into neutrophil-like cells under specified experimental conditions. The 5×104 cells that received Indirubin treatment and subsequent washing were placed into black clear-bottomed 96-well plates and preincubated with Fluo-3 reagent from the Calcium Assay Kit and 2×5mmol/L probenecid to block dye release at 37°C for 1 hour. The calcium response experiments utilized an integrated fluid transfer workstation operating at 37°C with an excitation wavelength of 485nm and an emission wavelength of 525nm. Record data as relative fluorescence units (RFU) at 2-second intervals. A dye-exclusion test with trypan blue examined cell viability. |
Reaction Conditions | 1μmol/L; 6 days |
Applications | Indirubin enhanced superoxide production by terminally differentiated HL-60 cells in the presence of opsonized zymosan, and enhanced the neutrophilic differentiation of HL-60 cells. |
| Animal experiment [2]: | |
Animal models | Kunming pregnant mice |
Preparation Method | Thirty-six Kunming pregnant mice were randomly divided into six groups: the experimental groups included a control group and subsequent groups treated with dimethyl sulfoxide (DMSO), LPS alone, and LPS with Indirubin at concentrations of 25μmol/L, 50μmol/L, and 100μmol/L, respectively, for 24 hours. The mice underwent separation from their offspring two hours before receiving an anesthetic injection of pentobarbital sodium salt which consisted of 0.25g dissolved in 50mL phosphate-buffered saline (PBS) and administered at 10 mg per 20g intraperitoneally (i.p.). LPS groups were used as positive controls. The research group administered various Indirubin doses(25-100μmol/L) and 0.1% DMSO via intraperitoneal injection one hour before and twelve hours after LPS administration. Both the control group and LPS group received the same PBS volume. The mice received CO2 euthanasia after 24 hours of LPS challenge before collecting mammary gland tissue to store at −80°C using a cell-freezer storage tube freezer. Measure the concentrations of IL-1β, IL-6 and TNF-α in mammary gland tissue through ELISA according to the kit provider's guidelines. Collect mammary gland tissues for histopathological analysis 24 hours after the LPS challenge. Research required tissue samples to undergo fixation in 4% paraformaldehyde for 48-72 hours followed by dehydration with graded alcohol and embedding in paraffin before staining with hematoxylin and eosin. |
Dosage form | 25μmol/L, 50μmol/L, and 100μmol/L for 24h; i.p. |
Applications | Indirubin (100μmol/L) attenuated histopathological changes in the mammary gland, local tissue necrosis, and neutrophil infiltration in the LPS-induced mastitis mouse model. |
References: | |
| Cas No. | 479-41-4 | SDF | |
| 别名 | 靛玉红; Couroupitine B; Indigo red; Indigopurpurin | ||
| Canonical SMILES | O=C1/C(NC2=CC=CC=C21)=C3C4=CC=CC=C4NC\3=O | ||
| 分子式 | C16H10N2O2 | 分子量 | 262.26 |
| 溶解度 | DMF>1 mg/ml, DMSO>10 mg/ml | 储存条件 | Store at -20°C |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
||
| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
 |
1 mg | 5 mg | 10 mg |
| 1 mM | 3.813 mL | 19.065 mL | 38.1301 mL |
| 5 mM | 0.7626 mL | 3.813 mL | 7.626 mL |
| 10 mM | 0.3813 mL | 1.9065 mL | 3.813 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet