产品编号：H10027

产品名称：HA14-1

CAS:

65673-63-4

储存条件

粉末

-20℃

四年

分子式：

C17H17BrN2O5

溶于液体

-80℃

六个月

分子量

409.23

-20℃

一个月

化学名： 

Solubility (25°C)

体外

DMSO

82mg/mL (200.37mM)

Ethanol

82mg/mL (200.37mM)

Water

Insoluble

体内（现配现用）

1% DMSO+30% polyethylene glycol+1% Tween 80

30mg/mL

＜1mg/ml表示微溶或不溶。

普西唐提供的所有化合物浓度为内部测试所得，实际溶液度可能与公布值有所偏差，属于正常的批间细微差异现象。

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。

浓度

溶液体积

质量

1mg

5mg

10mg

1mM

2.4436mL

12.2181mL

24.4361mL

5mM

0.4887mL

2.4436mL

4.8872 L

10mM

0.2444mL

1.2218mL

2.4436mL

50mM

0.0489mL

0.2444mL

0.4887mL

产品描述

一种细胞渗透性，非肽类Bcl-2抑制剂，IC5：9μM左右。

靶点/IC50

Bcl-2

9μM (IC50)

Bcl-xL

体外研究

HA14-1 is a nonpeptidic ligand of a Bcl-2 surface pocket. HA14-1 induces the activation of Apaf-1 and caspases, possibly by binding to Bcl-2 protein and inhibiting its function. The interaction of HA14-1 with the Bcl-2 surface pocket appeared to be specific for the chemical structure of HA14-1 as a series of synthetic analogs derived from HA14-1 containing various modifications are found to have widely different Bcl-2 binding activities. To examine the effect of HA14-1 on cell viability, HL60 cells are treated with various concentrations of HA14-1 for 4h. HA14-1 induces the death of HL-60 cells in a dosedependent manner. At 50μM, HA14-1 causes the lost of viability in more than 90% of the cells. HA14-1 is a Bcl-2/Bcl-xL antagonist.Swiss nude mice are challenged with BeGBM cells (104 injected s.c.).HA14-1 (400nmol) in 100μL free RPMI 1640-50% DMSO,or the carrier alone, is given at the site of injection once weekly from day 2 following cell injection. HA14-1 treatment does not have any significant effect on the growth of glioblastoma tumors in immunodeficient mice as monitored twice weekly by measuring the volume of the expanding tumors. Moreover, no gross organ toxicity or weight loss can be detected in mice receiving such treatment. To analyze whether HA14-1 treatment might enhance the efficiency of another antitumoral treatment, Swiss nude mice challenged with human glioblastoma multiforme cells are also given i.p. low doses of Etoposide (2.5mg/kg in 200μL of 0.9% NaCl 5 days a week from day 2 following cell injection) together with HA14-1 or mock treatment.Whereas Etoposide treatment is insufficient by itself to restrain the growth of glioblastoma cells, its combination with HA14-1 leads to a significant restrain on tumor growth as judged by the ability of the combined treatment to increase the doubling time of the tumor volume.

体内研究

Swiss nude mice are challenged with BeGBM cells (104 injected s.c.).HA14-1 (400nmol) in 100μL free RPMI 1640-50% DMSO,or the carrier alone, is given at the site of injection once weekly from day 2 following cell injection. HA14-1 treatment does not have any significant effect on the growth of glioblastoma tumors in immunodeficient mice as monitored twice weekly by measuring the volume of the expanding tumors. Moreover, no gross organ toxicity or weight loss can be detected in mice receiving such treatment. To analyze whether HA14-1 treatment might enhance the efficiency of another antitumoral treatment, Swiss nude mice challenged with human glioblastoma multiforme cells are also given i.p. low doses of Etoposide (2.5mg/kg in 200μL of 0.9% NaCl 5 days a week from day 2 following cell injection) together with HA14-1 or mock treatment.Whereas Etoposide treatment is insufficient by itself to restrain the growth of glioblastoma cells, its combination with HA14-1 leads to a significant restrain on tumor growth as judged by the ability of the combined treatment to increase the doubling time of the tumor volume.

激酶实验：

The binding affinity of organic compounds to Bcl-2 protein in vitro is determined by a competitive binding assay based on fluorescence polarization. For this assay, 5-carboxyfluorecein is coupled to the N terminus of a peptide,GQVGRQLAIIGDDINR, derived from the BH3 domain of Bak (Flu-BakBH3), which has been shown to bind to the surface pocket of the Bcl-xLprotein with high-affinity (dissociation constant, KD, of ≈0.34μM). According to our molecular modeling studies and binding measurement using fluorescence polarization,the Flu-BakBH3 peptide binds the surface pocket of Bcl2 with a similar affinity (dissociation constant, KD, of ≈0.20μM). Bcl-2 used in this assay is a recombinant GST-fused soluble protein. Flu-BakBH3 and Bcl-2 protein are mixed in the presence or absence of organic compounds under standard buffer conditions and are incubated for 30 min. The binding of Flu-BakBH3 to Bcl-2 protein is measured on a LS-50 luminescence spectrometer equipped with polarizers using a dual path length quartz cell (500μL). The fluorophore is excited with vertical polarized light at 480nm (excitation slit width 15nm), and the polarization value of the emitted light is observed through vertical and horizontal polarizers at 530nm (emission slit width 15nm). The binding affinity of each compound for Bcl-2 protein is assessed by determining the ability of different concentrations of the compound to inhibit Flu-BakBH3 binding to Bcl-2.

细胞实验:

The dose-dependent effect of the compounds on the viability of HL-60 cells is tested by using the CellTiter 96AQ kit. In brief,the cell suspension containing 1×105 cells in 100μL of medium are plated into a 96-well plate and are incubated with compounds (e.g., HA14-1;10, 20, 30, 40, 50 and 60μM) at different concentrations. The numbers of apoptotic cells are determined by measuring the optical density on a Wallac Victor counter at 490 nm. Cell viability is also determined by Trypan blue exclusion in a hemocytometer.

动物实验:

Mice

Six- to 7-week-old female Swiss nude mice are maintained under standard conditions for 2 weeks before their s.c. injection with 104 BeGBM cells (resuspended in 100μL free RPMI 1640) into the hind limbs on day 0. Treatment with HA14-1 is done by injection of 400nmol HA14-1 in 100μL free RPMI 1640-50% DMSO at the site of cell injection. Treatment starts at day 2 and is repeated once weekly for the following 4 weeks. Control mice receive equivalent volumes of vehicle (100μL free RPMI 1640-50% DMSO) with the same frequency.Etoposide treatment is done as follows: either Etoposide (2.5mg/kg) or sterile physiologic saline is given i.p. to HA14-1- or mock-treated animals on days 2 and 3 followed by five injections per week during the next 4 weeks. Tumor sizes are measured with calipers by the same observer twice weekly. Tumor volumes are calculated.