体外研究
(In Vitro) |
Tubulin polymerization-IN-6 (compound 5f) (0-20 μM, 24 h) shows a broad spectrum of anti-proliferation activity against cancer cell lines[1].
Tubulin polymerization-IN-6 (0-100 nM, 24 h) inhibits tumor cells colony formation, up-regulates the expression of Ac-α-tubulin and DeY-α-tubulin [1].
Tubulin polymerization-IN-6 (0-5 μM, 1 h) competes with colchicine and directly binds to the colchicine binding site, thus inhibit tubulin polymerization[1].
Tubulin polymerization-IN-6 (0-250 nM, 24 h) possesses a favorable anti-migration activity against cancer cells[1].
Tubulin polymerization-IN-6 (0-50 nM, 24 h) has the ability to inhibit the angiogenesis of HUVEC cells[1].
Tubulin polymerization-IN-6 (0-100 nM, 24 h) induces cell cycle arrest by regulating associated proteins, induces apoptosis by regulating associated proteins and down-regulating mitochondrial membrane potential, and dose-dependently promotes the production of ROS in HT29 cells[1].
Shanghai Jinpan Biotech Co Ltd has not independently confirmed the accuracy of these methods. They are for reference only.
Cell Proliferation Assay
| Cell Line: |
HT29, MCF-7, HeLa, MDA-MB-231, A549[1] |
| Concentration: |
0-20 μM |
| Incubation Time: |
24 h |
| Result: |
Had a broad spectrum of anti-proliferation activity against cancer cell lines (MCF-7, MDA-MB-231, A549, Hela, and HT29), with IC50 values of 0.14 ± 0.03, 0.10 ± 0.00, 0.24 ± 0.03, 0.035 ± 0.002, and 0.023 ± 0.001 μM, respectively; and showed moderate anti-proliferative activity against drug resistant cancer cells (MCF-7/TxR and A549/TxR), with IC50 values of 0.18 ± 0.02 and 0.31 ± 0.08 μM, and DRI (drug-resistant index) of 1.3 and 1.2, respectively. |
Western Blot Analysis
| Cell Line: |
HT29 cells[1] |
| Concentration: |
0, 25, 50, and 100 nM |
| Incubation Time: |
24 h |
| Result: |
Up-regulated the expression of Ac-α-tubulin (acetyl-α-tubulin) and DeY-α-tubulin (detyrosinated-α-tubulin); regulated the expressions of the proteins involved in cell cycle such as cdc25c, cdk7, cyclin B1, and cdc2; down-regulated the level of Bim and up-regulated the levels of Bcl-2, p-Bcl-2, and Bax, decreased the expression of p-Histone H3(Ser10) and increased the expression of cleaved-Caspase-9, cleaved-Caspase-3, PARP, and cleaved-PARP. |
Immunofluorescence
| Cell Line: |
HT29 cells[1] |
| Concentration: |
0, 25, 50, and 100 nM |
| Incubation Time: |
6 h |
| Result: |
Dose-dependently depolymerized the tubulin polymers into oligomers, and caused the microtubule network to collapse in HT29 cells. |
Cell Cycle Analysis
| Cell Line: |
HT29 cells[1] |
| Concentration: |
0, 12.5, 25, 50, and 100 nM |
| Incubation Time: |
24 h |
| Result: |
Induced a dose dependent G2/M phase arrest, increased the proportion of G2/M phase cells from 20.9% to 87.5% at 100 nM. |
Apoptosis Analysis
| Cell Line: |
HT29 cells[1] |
| Concentration: |
0, 25, 50, and 100 nM |
| Incubation Time: |
24 h |
| Result: |
Induced apoptosis, increased the percentages of total apoptosis cells, down-regulated mitochondrial membrane potential. |
|
体内研究
(In Vivo) |
Tubulin polymerization-IN-6 (compound 5f) (HT29 xenograft Balb/c nude mice, 0-10 mg/kg, IP, once every two days, for three weeks) dose-dependently inhibits the tumor growth[1].
Tubulin polymerization-IN-6 (SD rats, 10 mg/kg, IV, once) shows the better pharmacokinetic properties[1]. Pharmacokinetic Parameters of Tubulin polymerization-IN-6 in SD rats[1].
| Parameters |
5f |
| t1/2 (h) |
1.73 |
| AUC (μg/L·h) |
5.67 |
| MRT (h) |
1.92 |
| CL (L/h/kg) |
1.76 |
| Tmax (h) |
0.14 |
| Cmax (ng/mL) |
6.88 |
Shanghai Jinpan Biotech Co Ltd has not independently confirmed the accuracy of these methods. They are for reference only.
| Animal Model: |
Immunodeficient Balb/c nude mice (HT29 xenograft, 5-week-old, 36 mice, six groups)[1] |
| Dosage: |
0, 5, 7.5, 10 mg/kg |
| Administration: |
IP, once every two days, for three weeks |
| Result: |
Dose-dependently inhibited the tumor growth, inhibits the tumor weight growth by 75.5% at 10 mg/kg. |
| Animal Model: |
SD rats (5-week-old)[1] |
| Dosage: |
10 mg/kg |
| Administration: |
IV, once (Pharmacokinetic Analysis) |
| Result: |
Showed the better pharmacokinetic properties, exhibited an eight-fold half-life and a two-fold AUC improvement. |
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