甲异靛联合imatinib对CML K562细胞的促凋亡作用

目的研究甲异靛联合imatinib促进K562细胞凋亡的作用。方法K562细胞经imatinib或/和甲异靛处理48h后,采用流式细胞术检测细胞凋亡相关指标(线粒体跨膜电位和AnnexinV/PI)的改变,ELISA方法检测caspase3活性改变,Westernblot检测凋亡相关蛋白(cytC,cIAP1,procaspase3和PARP)的改变。结果与单用组相比,20μmol/L甲异靛与0.3μmol/Limatinib联合应用后,线粒体跨膜电位下降细胞比例明显升高,cytC释放入胞浆,caspase3酶原形式procaspase3减少,caspase3活性升高,其底物PARP被降解,AnnexinV水平升高。结论甲异靛与imatinib合用可协同促进K562细胞凋亡。

Imatinib治疗转移性、不能手术切除的胃肠间质瘤

1文献类型治疗2证据水平2b3文献来源Demetri G D, Mehren M V, Blanke C D, et al.Efficacy and safety of imatinib mesylate in advanced gastrointestinal stromal tumors [J]. N Engl J Med,2002。

动力学方法研究Imatinib与蛋白激酶的结合构象与结合自由能

对imatinib两种典型复合物晶体结构imatinib-Abl与imatinib-c-Kit进行比对分析,并利用MM/PBSA方法计算imatinib与Abl、c-Kit、c-Src、p38及Syk等激酶的结合自由能.计算结果表明,imatinib与上述激酶结合时,范德华作用对结合自由能有主要贡献;其中,imatinib与激酶c-Kit结合自由能最低;imatinib与非活性构象激酶的结合自由能明显低于与活性构象激酶的结合自由能.

胃肠间质瘤患者中Imatinib血药浓度与治疗疗效的相关性

目的检测中国晚期胃肠间质瘤(GIST)患者中伊马替尼(Imatinib)血药浓度,并分析血药浓度与治疗疗效的相关性。方法收集112例接受Imatinib治疗超过28 d的GIST患者的117份血浆样本,用HPLC-MS/MS法检测血浆中Imatinib浓度。分析Imatinib血药浓度与治疗疗效的相关性。结果高剂量组(600 mg/d)患者的血药浓度较低剂量组(400 mg/d)明显升高(P<0.001)。服药时间为1～2年患者的血药浓度(平均值1 268 ng/mL)较服药时间小于1年的患者(平均值1 845 ng/mL)明显降低(P<0.05)。随着血药浓度的增加有效率也随之增加,血药浓度低、中及高水平组的有效率分别为50%、67.5%及66.7%。结论 Imatinib血药浓度与服药剂量及服药时间明显相关;Imatinib治疗有效率随着血药浓度的增高呈增加趋势。

imatinib对大鼠SAH后早期颅脑损伤的影响

目的研究imatinib对蛛网膜下腔出血后早期颅脑损伤的作用及相关机制。方法收集160只Wistar大鼠,先将98只大鼠分为假手术组(n=20)、SAH组(n=26)、SAH+imatinib组(n=26)、SAH+imatinib+LY294002组(n=26)。采用血管内穿刺法制作SAH模型,假手术组不刺破血管,其余操作相同。再将30只SAH大鼠分为control si RNA组(n=10)、c-Abl si RNA组(n=10)和c-Abl si RNA+imatinib组(n=10)。计算各组大鼠的病死率,采用Western blot检测p-PDGFRα、c-Abl、p-Akt、p-GSK3β蛋白水平,Tunel荧光法检测神经元凋亡,评价出血后24、72 h各组大鼠SAH严重程度、神经评分;另取32只大鼠用于检测出血后48 h脑组织含水量,伊文思蓝检测血-脑屏障通透性变化。结果 imatinib显著降低出血后24 h病死率及神经元凋亡,减轻脑水肿,减少出血后48 h伊文思蓝渗出,提高72 h后神经评分(P <0.05),但不减少出血量。出血24 h后pPDGFRα、c-Abl蛋白表达显著增加,p-Akt、p-GSK3β表达稍增加(P <0.05),imatinib显著下调p-PDGFRα、c-Abl表达,上调p-Akt、p-GSK3β的表达(P <0.05),LY294002可拮抗imatinib作用(P <0.05)。沉默c-Abl基因表达,p-GSK3β、p-Akt的表达显著增加,神经元凋亡明显减少(P <0.05),在此基础上使用imatinib对p-GSK3β、p-Akt蛋白表达及凋亡无影响(P>0.05)。结论 imatinib减轻SAH早期颅脑损伤,改善神经功能,可能与抑制c-Abl、增加Akt/GSK3β磷酸化有关。

Effect of PDGF-Rb antagonist imatinib on endometrial injury repairing in mouse model

Objective: To study the effects of PDGF-Rb antagonists imatinib on endometrial injury repairing in the mouse model. Methods: The cultured MSCs cells from male mice were marked with Brd U in vitro, and then transplanted to the female mice which suffered from radiation injury through tail vein, PDGF-Rb antagonists imatinib was injected through abdominal cavity. Four groups were arranged, which were radiation transplantation group, normal control group, imatinib intervention group and radiation control group. Brd U incorporation, SRY expression and MVD status were detected in uterus of mice. Results: SRY gene was negative expressed in normal control group and radiation control group. SRY gene presented positive in radiation transplantation group and imatinib intervention group; Brd U incorporation showed negative in radiation control group and normal control group which died in the early stage in mice; the incorporation of Brd U was higher in radiation transplantation group compared with imatinib intervention group; CD34 was positive on the uterus of all the four groups,which showed highest in radiation control group and lowest in radiation control group; The MVD in imatinib intervention group was lower than radiation control group; the difference of MVD was significantly compared with normal control group(P<0.05). Conclusions: PDGF-Rb antagonists imatinib could inhibit the repairing function of MSCs in the endometrial lesions in mice.

A validated UPLC–MS/MS method for simultaneous determination of imatinib, dasatinib and nilotinib in human plasma

A sensitive, rapid, simple and economical ultra-performance liquid chromatography-tandem mass spectrometric method （UPLC-MS/MS） was developed and validated for simultaneous determination of imatinib, dasatinib and nilotinib in human plasma using gliquidone as internal standard （IS）. Liquid-liquid extraction method with ethyl acetate was used for sample pre-treatment. The separation was performed on an Xtimate Phenyl column using isocratic mobile phase consisting of A （aqueous phase： 0.15% formic acid and 0.05% ammonium acetate） and B （organic phase： aeetonitrile） （A：B=40：60, v/v）. The flow rate was 0.25 mL/min and the total run time was 6 min. The multiple reaction monitoring （MRM） transitions, m/z 494.5-394.5 for imatinib, 488.7-401.5 for dasatinib, 530.7-289.5 for nilotinib and 528.5-403.4 for IS, were chosen to achieve high selectivity in the simultaneous analyses. The method exhibited great improvement in sensitivity and good linearity over the concentration range of 2.6-5250.0 ng/mL for imatinib, 2.0-490.0 ng/mL for dasatinib, and 2.4-4700.0 ng/mL for nilotinib. The method showed acceptable results on sensitivity, specificity, recovery, precision, accuracy and stability tests. This UPLC-MS/MS assay was successfully used for human plasma samples analysis and no significant differences were found in imatinib steady-state trough concentrations among the SLC22A5 -1889T 〉 C or SLCOIB3 699G 〉 A genotypes （P 〉 0.05）. This validated method can provide support for clinical therapeutic drug monitoring and pharmacokinetic investigations of these three tyrosine kinase inhibitors （TKIs）.

Imatinib as preoperative therapy in Chinese patients with recurrent or metastatic GISTs

Objective： Imatinib has dramatically altered the options for management of patients with gastrointestinal stromal turnouts. However, it has become clear that secondary resistance to the drug develops during long- term therapy. The purpose of our study was to retrospectively analyze safety and long-term outcomes in Chinese patients with recurrent or metastatic GISTs treated with imatinib preoperatively. Methods： Between June 2003 and June 2011, 22 patients underwent surgery for recurrent or metastatic GISTs after preoperative treatment with imatinib. Results： Complete resection was accomplished in 8 of the 10 responsive disease （RD） patients （80%）, and in 3 of the 12 patients （25%） who had progression disease （PD）. The amount of blood loss during the operation in PD patients was higher than in RD patients. There was 1 hospital death in PD group related to surgery, while the other patients recovered with conservative therapy because complications were mild. The difference in median PFS between patients with RD and those with PD was significant （24.8 vs. 2.81 months, P〈0.001）. The difference in 2-year OS rate between patients with RE） and those with PD was not significant （100% vs. 87.5%, P〉0.05）. Conclusions： Our study indicates that surgical intervention can improve the PFS of Chinese patients with recurrent or metastatic GISTs responsive to imatinib, but does not prolong OS as well as in patients who develop imatinib resistance. Surgical resection following imatinib treatment is feasible and can be considered for patients with advanced GISTs responsive to imatinib.

光谱及分子模拟法对HSA和BSA与酪氨酸激酶抑制剂Imatinib相互作用的比较分析

研究一种酪氨酸激酶抑制剂(tyrosine kinase inhibitor,TKI)伊马替尼(imatinib,IMA)与人血清清蛋白(HSA)及牛血清清蛋白(BSA)的相互作用,比较分析HSA和BSA与IMA相互作用机制的差异.模拟生理条件下,计算机模拟技术结合荧光光谱和紫外光谱法,研究IMA与蛋白质的作用机制.分子模建IMA与血清清蛋白的结合模型,表明伊马替尼与蛋白质的相互作用力为疏水作用力,兼有氢键作用.光谱结果表明,IMA与HSA和BSA的相互作用表现为静态结合过程,结合强度较强,IMA与HSA和BSA分子的结合距离r值较小,说明发生了能量转移现象.IMA对HSA和BSA的结构域微区构象产生影响,使结合位域的疏水性发生改变.荧光相图技术解析出IMA与HSA和BSA反应构象型态的变迁为"二态"模型.HSA与IMA相互作用的热力学参数表明,IMA与HSA之间是以疏水作用为主的分子间作用,而IMA与BSA之间的作用力为氢键和范德华力,兼有少量的疏水作用力.光谱实验与计算机模拟结果基本一致,可为研究IMA与HSA和BSA相互作用本质提供一定参考.

Imatinib mesylate in clinically suspected gastric stromal tumors

Gastrointestinal stromal tumors （GISTs） occur most frequently in the stomach.Diagnosis of gastric GIST is not always clear before surgery.Flexible endoscopy may suggest the nature of the lesion （a bulky tumor with preserved mucosa）; however,biopsy is rarely diagnostic.Therefore,diagnostic medication with safe drugs may provide a feasible way under such conditions after an informed consent is obtained.Based on the excellent efficacy of imatinib mesylate （IM） in the treatment of GIST,we successfully applied it in the diagnostic medication of two patients with clinically suspected gastric stromal tumors.In conclusion,the diagnostic medication with IM can be an alternative option for patients with suspected GIST that can not be confirmed pathologically.

Imatinib response of gastrointestinal stromal tumor patients with germline mutation on KIT exon 13:A family report

Familial gastrointestinal stromal tumor(GIST) is a rare autosomal dominant disorder associated with mutations in the KIT gene in the majority of cases. Although, exon 11 appears to be the hot spot region for approximately 95% of germline mutations, pathogenic variations have also been identified in exon 8, 13 and 17. Exon 13 germline mutations are extremely rare amongst familial GISTs and seven families with a germline mutation have been reported to date. Moreover, the role of imatinib mesylate in this rare familiar settings is not completely known so far. We describe here clinical, imaging, pathological and genetic findings of a family with four affected members; grandmother, his son and two grand-sons having a germline gain-of-function mutation of KIT in exon 13 and discuss the imatinib mesylate treatment surveillance outcomes towards disease management.

Criminal or bystander: imatinib and second primary malignancy in GIST patients

Niigata University Hospital is a regional center institution of cancer therapy where many patients with gastrointestinal stromal tumors （GISTs） are visiting to seek the latest treatment.During the time Ⅰ was treating GIST patients there with imatinib,a tyrosine kinase inhibitor,a small concern was raised：Ⅰ successively encountered patients who were newly diagnosed as having malignant neoplasms during the course of their treatment.Of the 70 GIST patients who were enrolled in our prospective study of imatinib therapy,seven suffered from second primary malignancies （SPMs）.One female GIST patient who suffered from advanced esophageal cancer died of the SPM,whereas the remaining six patients continued with their imatinib therapy and their prognoses were not affected by their SPMs.I reported on the risk of SPMs in GIST patients under imatinib therapy to an international journal of clinical oncology （1）.As the patient cohort of our study was so small in number to apply to statistical analysis,our observation was no more than a clinical alert.

Evaluation of imatinib mesylate(Gleevec) on KAI1/CD82 gene expression in breast cancer MCF-7 cells using quantitative real-time PCR

Objective: To evaluate the effect of imatinib mesylate on cell viability, anti cancer effect through modulation of KAI1/CD82 gene expression in breast cancer MCF-7 cell line.Methods: The effects of imatinib mesylate on cell viability in MCF-7 cell line were assessed using MTT assay and IC_(50) value was determined. GAPDH and KAI1/CD82 were selected as reference and target genes, respectively. Quantitative real time PCR technique was applied for investigation of KAI1/CD82 gene expression in human breast cancer MCF-7 cells. Subsequently, the quantity of KAI1 compared to GAPDH gene expressions were analyzed using the formula; 2^(-DDCt).Results: Imatinib was showed to have a dose-dependent inhibitory effect on the viability of MCF-7 cells. CD82/GAPDH gene expression ratios were 1.322 ± 0.030(P > 0.05),2.052 ± 0.200(P < 0.05), 2.151 ± 0.270(P < 0.05) for 10, 20 and 40 mmol/L of imatinib concentrations.Conclusions: Based on the present data, imatinib mesylate might modulate metastasis by up-regulating KAI1/CD82 gene expression in human breast MCF-7 cancer cell line.

Idiopathic hypereosinophilic syndrome presenting with severe vasculitis successfully treated with imatinib

Idiopathic hypereosinophilic syndrome(HES) is a rare disorder characterized by peripheral eosinophilia exceeding 1500/mm3, a chronic course, absence of secondary causes, and signs and symptoms of eosinophil-mediated tissue injury. One of the best-characterized forms of HES is the one associated with FIP1L1-PDGFRA gene rearrangement, which was recently demonstrated as responsive to treatment with the small molecule kinase inhibitor drug, imatinib mesylate. Here, we describe the case of a 51-year-old male, whose symptoms satisfied the clinical criteria for HES with cutaneous and cardiac involvement and who also presented with vasculitic brain lesions and retroperitoneal bleeding. Molecular testing, including fluorescence in situ hybridization, of bone marrow and peripheral blood showed no evidence of PDGFR rearrangements. The patient was initially treated with high-dose steroid therapy and then with hydroxyurea, but proved unresponsive to both. Upon subsequent initiation of imatinib mesilate, the patient showed a dramatic improvement in eosinophil count and progressed rapidly through clinical recovery. Long-term follow-up confirmed the efficacy of treatment with low-dose imatinib and with no need of supplemental steroid treatment, notwithstanding the absence of PDGFR rearrangement.

Repositioning imatinib for spinal cord injury

Efforts to pharmacologically restore central nervous system（CNS）function after injury has historically focused on promoting nerve growth with nerve growth factors such as nerve growth factor（NGF）,neurotrophin 3（NT3）and brain-derived neurotrophic factor（BDNF）.

GIST中imatinib耐药机制研究进展

在功能获得性突变所致的KIT或PDGFRA受体酪氨酸激酶异常活化与胃肠间质瘤(GIST)的发病关系阐明后,酪氨酸激酶抑制剂甲璜酸伊马替尼(imatinib)在进展期GIST中显现出很高的有效性,使GIST的治疗发生了革命性的改变。然而,GIST中对imatinib的早期或晚期耐药已经成为日益严重的临床问题。因此,对耐药的分子机制的进一步了解就成为当前研究的焦点,本文综述了目前对imatinib耐药的分子机制的认识,这些认识可能引导新的治疗策略的产生。

Gastrointestinal stromal tumors and second primary malignancies before and after the introduction of imatinib mesylate

Gastrointestinal stromal tumor （GIST） is the most common mesenchymal malignancy of the gastrointestinal tract.GISTs may coexist with different types of cancer,either synchronous or metachronous （1）.Most GISTs develop in a sporadic fashion,but familial occurrence,such as neurofibromatosis and Carney-triad,has also been reported （2）.The overall frequency of second tumors in different series varied from 4.5% to 33%.The most frequent types of GIST-associated cancers were gastrointestinal carcinomas （47%）,lymphoma/leukemia （7%）,carcinomas of prostate （9%）,breast （7%）,kidney （6%）,lung （5%）,female genital tract （5%）,carcinoid tumors （3%）,soft tissue and bone sarcomas （3%）,malignant melanoma （2%） and seminoma （1%） （1,3-5）.

Characterization of Genomic Events Other than Ph and Evaluation of Prognostic Influence on Imatinib in Chronic Myeloid Leukemia (CML): A Study on 1449 Patients from India

Background: Analysis of Philadelphia (Ph) chromosome, a hallmark of chronic myeloid leukemia (CML) plays an important role in disease monitoring of the targeted drug Imatinib. Apart from Ph, genomic imbalances such as additional chromosomal abnormalities (ACAs) of major route occur during transformation of the disease and show negative impact on prognosis. Objective: The present study was carried out to investigate frequencies of ACAs, genomic deletions, complex Ph variants and their prognostic influences in a large cohort of newly diagnosed CML-CP (chronic phase) and CML-AP/BP (accelerated/blast phase). Material & Methods: Retrospective, single institutional study on 1367 cases of CML-CP and 82 cases of CML-AP/BP between 2009 and 2015, using conventional cytogenetics along with fluorescence in situ hybridization. Results: Of the 1367 patients in CML-CP, 1041 patients who completed 12 - 18 months of Imatinib therapy showed complete cytogenetic remission (CCyR) rates of 76% and 82% at 12 and 18 months respectively. Imatinib induced 81% and 33% CCyR in CML-AP and CML-BP respectively. Frequencies of ACAs in CML-CP, AP and BP were 2%, 27% and 67% respectively. Patients in chronic and AP/BP phase with ACAs showed resistance to Imatinib (p < 0.0005). The incidence of genomic deletions and complex Ph variants was 21% and 6.3% respectively with no comparable difference of cytogenetic response to Imatinib (p p < 0.210 respectively). In a cohort of 112 patients in CCyR, development of new clonal abnormalities, more frequently trisomy 8 was detected in Ph negative clone. Conclusion: Our data demonstrated that Imatinib as a frontline therapy had significantly improved management of CML. However, ACAs play an important role in resistance to Imatinib, both in chronic and acute phase, which may limit sole ABL targeted therapy.

Bilateral Masculine Mastoplasia Associated with Imatinib Mesylate:A Case Report and Literature Review

1 CASE REPORT In June 2009, a 29-year-old Chinese male was diagnosed as having Philadelphia chromosome-positive chronic myeloid leukemia （chronic phase）; other than a high white blood cell count in peripheral blood （WBC, 254.00×10^9/L） and splenomegaly, the patient exhibited no abnormal physical signs in mammary glands. He was given hydroxyurea for several days before he received treatment with 400 mg of imatinib mesylate daily.

Raman spectroscopy for detection of imatinib in plasma: A proof of concept

Imatinib is the standard first line treatment for chronic myeloid leukemia(CML).Owing to doserelated toxicities of Imatinib such as neutropenia,there is scope for treatment optimization through therapeutic drug monitoring(TDM).Trough concentration of 1
g/mL is considered the therapeutic threshhold.Existing methods for the detection of Imatinib in plasma are limited by long read out time and expensive instrumentation.Hence,Raman spectroscopy was explored as a rapid and objective tool for monitoring Imatinib concentration.Three approaches:conventional Raman spectroscopy(CRS),Drop coating deposition Raman(DCDR)spectroscopy and surface-enhanced Raman spectroscopy(SERS)were employed to detect the required trough concentration of 1
g/mL and above.Detection of therapeutically relevant concentrations(1
g/mL)using SERS and suitable nanoparticle substrates has been demonstrated.Prospectively,rigorous validation using clinical samples is necessary to confirm the utility of this approach in routine clinical usage.