伊马替尼治疗失败慢性期CML患者及早干预的意义及二代TKIs选择

慢性髓细胞性白血病(CML)应用伊马替尼后,明显改善了预后,但治疗失败的患者则预后不良。为改善这些患者的预后,一方面需要加强细胞遗传学和分子生物学监测,及早发现治疗失败并给予干预。另一方面,对于大多数治疗失败的患者,二代酪氨酸激酶抑制剂(TKIs)是合适的选择,如果就目前临床上两种二代TKIs进行比较,总体看来尼洛替尼好于达沙替尼。     

Curcumin derivative,C817 inhibits proliferation of imatinib-resistant chronic myeloid leukemia cells with wild-type or mutant Bcr-Abl in vitro

Aim： To find new kinase inhibitors that overcome time imatinib resistance in treatment of chronic myeloid leukemia （CML）, we synthesized C817, a novel derivative of curcumin, and tested its activities against wild-type （WT） and imatinib-resistant mutant Abl kinases, as well as in imatinib-sensitive and resistant CML cells in vitro. Methods： 32D cells harboring WT or mutant Abl kinases （nucleotide binding P-loop mutants Q252H, Y253F, and imatinib contact residue mutant T3151）, as well as K562/G01 cells （with whole Bcr-Abl gene amplication） were tested. Kinase activity was measured using Kinase-GIo Luminescent Kinase Assay Platform in recombinant WT and mutant （Q252H, Y253F, and T3151） Abl kinases. Cell proliferation and apoptosis were examined using MTT assay and flow cytometry, respectively. The phosphorylation levels of Bcr-Abl initiated signaling proteins were analyzed using Western blotting. Colony forming units （CFU） growth and long term culture-initiating cells （LTC-ICs） were used to test the effects of C817 on human leukemia progenitor/stem cells. Results： C817 potently inhibited both WT and mutant （Q252H, Y253F, and T3151） Abl kinase activities in a non-ATP competitive manner with the values of ICso at low nanomole levels. In consistent with above results, C817 suppressed the growth of both imatinib-sensitive and resistant CML cells, including wild-type K562, K562/GO$, 32D-T3151, 32D-Q252H, and 32D-Y253F cells with the values of ICso at low micromole levels. C817 （0.5 or 1 tJmol/L） dose-dependently inhibited the phosphorylation of Bcr-Abl and downstream proteins STAT-5 and CrkL in imatinib-resistant K562/G01 cells. Furthermore, C817 significantly suppressed CFU growth and LTC-ICs, implicating that C817 could eradiate human leukemia progenitor/stem cells. Conclusion： C817 is a promising compound for treatment of CML patients with Bcr-Abl kinase domain mutations that confer imatinib resistance.     

伊马替尼治疗慢性髓细胞白血病慢性期临床疗效观察

目的：分析甲磺酸伊马替尼（IM）治疗慢性髓系白血病（CML）慢性期的临床疗效及影响疗效的因素。方法随访观察74例 CML 慢性期患者,IM中位治疗剂量为400（200～600）mg·d －1,评估其临床疗效,总生存时间和疾病无进展生存时间,并对相关疗效影响因素进行分析。结果中位随访时间为20（6～72）个月,累积达到血液学缓解（CHR）为98．6％,血液学中位缓解时间1（1～3）月;63例（85．1％）达到主要细胞遗传学缓解（MCyR）,中位达 MCyR 时间为9（5～24）个月;53例（71．6％）达完全细胞遗传学缓解（CCyR）,41例（55．4％）达到主要分子生物学缓解（MMR）;6例（8．1％）达到完全分子生物学缓解（CMR）;初治组及复治组应用 IM治疗后 CHR 及 MCyR 差异无统计学意义,但 CCyR 及 MMR 差异均有统计学意义（P ＜0．016）;由 EUTOS 评分区分的低危组与高危组应用 IM治疗后 CHR 差异无统计学意义,但 MCyR、CCyR 及 MMR 差异均有统计学意义（P ＜0．020）;其中初治组与复治组及 EUTOS 评分低危组与高危组 OS 差异无统计学意义,但其 PFS 差异均有统计学意义（P 分别为0．021和0．004）。结论IM用于 CML 慢性期患者治疗可获得极高的血液学缓解率和较高细胞遗传学缓解率,不良反应少,提高了患者生存质量,延长患者的生存时间;在 CML 确诊早期应用可提高疗效,IM治疗前时间大于6个月或EUTOS 评分高危组可影响 IM疗效。     

伊马替尼与干扰素联合治疗慢性粒细胞白血病的疗效分析

目的:探讨伊马替尼与干扰素联合化疗治疗慢性粒细胞白血病(CM L)的疗效。方法:2004年6月—2009年7月新诊断的58例Ph染色体阳性CM L慢性期患者,随机分为伊马替尼组和干扰素联合化疗组,比较两组临床疗效。结果:两组总有效率差异无统计学意义(P>0.05);伊马替尼组完全血液学缓解率,完全细胞遗传学缓解率、完全分子学效应率、5年总生存率均明显高于干扰素联合化疗组(P<0.05)。结论:伊马替尼和干扰素联合化疗都可作为CM L慢性期的有效治疗方法,应依据不同情况实施个体化治疗。     

Imatinib plasma trough concentration and its correlation with characteristics and response in Chinese CML patients

Aim:

To investigate the pharmacokinetics of imatinib in Chinese chronic myelogenous leukemia (CML) patients.

Methods:

Fourty-six naïve Chinese CML patients treated with imatinib (400 and 600 mg daily, n=36 and 10, respectively) were recruited. The correlations of imatinib (400 mg) trough plasma concentrations (C_mins) with the patients'' characteristics and responses were analyzed.

Results:

The overall mean (±SD, CV%) steady-state C_mins for imatinib at 400 mg (n=36) and 600 mg (n=10) daily was 1325.61 ng/mL (±583.53 ng/mL; 44%) and 1550.90 ng/mL (±462.63 ng/mL; 30%), respectively, and no statistically significant differences were found between them (P=0.267). At 400 mg daily, female patients had significantly higher C_mins than the male patients (P=0.048), and molecular responses were not correlated with imatinib C_mins, but they were correlated with time elapsed before imatinib therapy.

Conclusion:

The results suggest that Chinese CML patients have higher imatinib C_mins than their Caucasian counterparts and that the optimal initial imatinib dose for them requires further investigation.     

GA3, a new gambogic acid derivative,exhibits potent antitumor activities in vitro via apoptosis-involved mechanisms

Aim： Gambogic acid （GA） is the major active ingredient of gamboge, which is secreted from a Chinese traditional medicine, Garcinia hanburyi, which possesses potent antitumor activity. GA3, a new GA derivative, has been shown to possess better water solubility than GA. The aim of the present study was to examine the antitumor activity of GA3 and the mechanism underlying it.
Methods： The growth inhibition of cancer cell lines induced by GA3 was assessed using the SRB assay. DAPI staining, flow cytometry, a DNA fragment assay, and Western blot analysis were used to study the apoptotic mechanisms of GA3.
Results： GA3 displayed wide cytotoxicity in diversified human cancer cell lines with a mean ICs0 value of 2.15 μmol/L. GA3 was also effective against multidrug resistant cells, with an average resistance factor （RF） that was much lower than that of the reference drug, doxorubicin. Mechanistic studies revealed that GA3-induced apoptosis in HL-60 cells proceeded via both extrinsic and intrinsic pathways, with caspase-8 functioning upstream of caspase-9. In addition, GA3-driven apoptotic events were associated with up-regulation of Bax, down-regulation of Bcl-2 and cleavage of Bid. Moreover, GA3 triggered cytochrome c release from the mitochondria, in particular bypassing the involvement of the mitochondrial membrane potential.
Conclusion： Better solubility and a potential anti-MDR activity, combined with a comparable antitumor efficacy, make GA3 a potential drug candidate in cancer therapy that deserves further investigation.     

伊马替尼治疗常规化学治疗失败的进展期慢性粒细胞白血病21例

目的:观察伊马替尼治疗常规化学治疗(化疗)失败的进展期慢性粒细胞白血病(CML)的有效性及不良反应。方法:21例病人在常规化疗无效后服用伊马替尼,其中加速期4例,急变期17例,起始剂量400 mg·d~(-1),无明显不良反应后改为600 mg·d~(-1)。结果:所有病人服用伊马替尼后,白细胞、血小板及幼稚细胞均明显下降,与服药前比较差异非常显著(P<0.01)。除1例因败血症死亡,2例病人骨髓部分缓解外,其余病人服药后7～45d均达到血液学完全缓解。伊马替尼对白细胞影响较明显,部分病人出现恶心、呕吐等消化道症状及眼睑水肿、肌痛等不良反应。结论:伊马替尼治疗常规化疗失败的难治性CML有较高的血液学缓解率,服用方便,效果好,不良反应轻。     

GA3, a new gambogic acid derivative,exhibits potent antitumor activities in vitro via apoptosis-involved mechanisms

Aim:

Gambogic acid (GA) is the major active ingredient of gamboge, which is secreted from a Chinese traditional medicine, Garcinia hanburyi, which possesses potent antitumor activity. GA3, a new GA derivative, has been shown to possess better water solubility than GA. The aim of the present study was to examine the antitumor activity of GA3 and the mechanism underlying it.

Methods:

The growth inhibition of cancer cell lines induced by GA3 was assessed using the SRB assay. DAPI staining, flow cytometry, a DNA fragment assay, and Western blot analysis were used to study the apoptotic mechanisms of GA3.

Results:

GA3 displayed wide cytotoxicity in diversified human cancer cell lines with a mean IC₅₀ value of 2.15 μmol/L. GA3 was also effective against multidrug resistant cells, with an average resistance factor (RF) that was much lower than that of the reference drug, doxorubicin. Mechanistic studies revealed that GA3-induced apoptosis in HL-60 cells proceeded via both extrinsic and intrinsic pathways, with caspase-8 functioning upstream of caspase-9. In addition, GA3-driven apoptotic events were associated with up-regulation of Bax, down-regulation of Bcl-2 and cleavage of Bid. Moreover, GA3 triggered cytochrome c release from the mitochondria, in particular bypassing the involvement of the mitochondrial membrane potential.

Conclusion:

Better solubility and a potential anti-MDR activity, combined with a comparable antitumor efficacy, make GA3 a potential drug candidate in cancer therapy that deserves further investigation.     

Some 2-[4-(1H-benzimidazol-1-yl) phenyl]-1H-benzimidazole derivatives overcome imatinib resistance by induction of apoptosis and reduction of P-glycoprotein activity

Imatinib (IMA) is a tyrosine kinase inhibitor (TKI) introduced for the chronic myeloid leukemia (CML) therapy. Emergence of IMA resistance leads to the relapse and failure in CML therapy. Benzimidazole is a heterocyclic organic compound which is widely investigated for the development of anticancer drugs. In this study, we aimed to explore the anticancer effects of some 2-[4-(1H-benzimidazol-1-yl) phenyl]-1H-benzimidazole derivatives on K562S (IMA-sensitive) and K562R (IMA-resistant) cells. To analyze the cytotoxic and apoptotic effects of the compounds, K562S, K562R, and L929 cells were exposed to increasing concentrations of the derivatives. Cytotoxic effects of compounds on cell viability were analyzed with MTT assay. Apoptosis induction, caspase3/7 activity were investigated with flow cytometry and BAX, BIM, and BAD genes expression levels were analyzed with qRT-PCR. Rhodamine123 (Rho-123) staining assays were carried out to evaluate the effect of compounds on P-glycoprotein (P-gp) activity. The hit compounds were screened using molecular docking, and the binding preference of each compounds to BCR-ABL protein was evaluated. Our results indicated that compounds triggered cytotoxicity, caspase3/7 activation in K562S and K562R cells. Rho-123 staining showed that compounds inhibited P-gp activity in K562R cells. Overall, our results reveal some benzimidazole derivatives as potential anticancer agents to overcome IMA resistance in CML.     

聚焦慢性粒细胞白血病

随着靶向治疗时代的到来,慢性粒细胞白血病（CML）已经从不治之症转变为基本可控的慢性病。患者生存率有了显著提高,当然在疗效、耐受性及耐药性方面仍有提升的空间。长期以来,酪氨酸激酶抑制剂格列卫（Gleevec）一直被认为是合理药物设计的典范,但更有效的二代药物已经开始作为一线药物获得认可。然而,由于缺乏完整的生存期数据,这些二代药物和格列卫相比所具去发现。由于患者需要长时间治疗,毒性和成本的可控性更可能成为选择治疗药物的重要推动因素。治疗慢性粒细胞白血病的产品线首先侧重于解决耐药性问题,尤其是在一线药物治疗失败而三线药物又无法满足需求的情况下。如果患者使用酪氨酸激酶抑制剂有效,那么最终的问题是患者是否可以通过这些药物治愈。     

DCLAK11, a multi-tyrosine kinase inhibitor,exhibits potent antitumor and antiangiogenic activity in vitro

Aim:

To investigate the molecular targets of DCLAK11, a novel compound discovered from a series of substituted pyridin-3-amine derivatives, and to characterize its anti-tumor properties in vitro.

Methods:

Kinase inhibition was measured by an ELISA assay. Cell viability was assessed with an SRB or a CCK8 assay. The alterations induced by kinase signaling proteins in cancer cells were detected by Western blot. Apoptosis was determined by an Annexin V-PI assay. The following assays were used to evaluate the impact on angiogenesis: wound-healing, Transwell, tube formation and microvessel outgrowth from rat aortic rings.

Results:

DCLAK11 was a multi-targeted kinase inhibitor that primarily inhibited the EGFR, HER2, and VEGFR2 tyrosine kinases with IC₅₀ value of 6.5, 18, and 31 nmol/L, respectively. DCLAK11 potently inhibited the proliferation of EGFR- and HER2-driven cancer cells: its IC₅₀ value was 12 and 22 nmol/L, respectively, in HCC827 and HCC4006 cells with EGFR exon deletions, and 19 and 81 nmol/L, respectively, in NCI-N87 and BT474 cells with HER2 amplification. Consistently, DCLAK11 blocked the EGFR and HER2 signaling in cancer cells with either an EGFR or a HER2 aberration. Furthermore, DCLAK11 effectively induced EGFR/HER2–driven cell apoptosis. Moreover, DCLAK11 exhibited anti-angiogenic activity, as shown by its inhibitory effect on the proliferation, migration and tube formation of human umbilical vascular endothelial cells and the microvessel outgrowth of rat aortic rings.

Conclusions:

DCLAK11 is a multi-targeted kinase inhibitor with remarkable potency against tyrosine kinases EGFR, HER2 and VEGFR2, which confirms its potent anti-cancer activity in EGFR- and HER2-addicted cancers and its anti-angiogenic activity.     

Dihydroartemisinin exhibits antitumor activity toward hepatocellular carcinoma in vitro and in vivo

Dihydroartemisinin (DHA), a semi-synthetic derivative of artemisinin isolated from the traditional Chinese herb Artemisia annua L., has been shown to exhibit inhibitory effects on human cancer cells. However, its antitumor ability toward hepatocellular carcinoma (HCC) has not been studied. In this study, we demonstrated that DHA significantly inhibited HCC cell growth in vitro and in vivo via inducing G2/M cell cycle arrest and apoptosis. The induction of p21 and the inhibition of cyclin B and CDC25C contributed to DHA-induced G2/M arrest. DHA-induced apoptosis was associated with mitochondrial membrane depolarization, release of cytochrome c, activation of caspases, and DNA fragmentation. Activation of caspase 9 and caspase 3, but not caspase 8, was detected in DHA-treated cells. Attenuation of apoptosis in cells pretreated with Z-VAD-FMK suggested the involvement of caspase cascade. Furthermore, p53 facilitated apoptosis caused by DHA. Bcl-2 family proteins were also responsible for DHA-induced apoptosis. DHA exposure decreased Mcl-1 expression but increased the levels of Noxa and active Bak. Bak was released from the Mcl-1/Bak complex due to the decline of Mcl-1. Further study revealed that Mcl-1 was rapidly degraded in DHA-treated cells and that DHA-induced apoptosis was largely inhibited by overexpression of Mcl-1 or RNAi-mediated decrease of Bak and Noxa. In a HCC-xenograft mouse model, the intraperitoneal injection of DHA resulted in significant inhibition of HCC xenograft tumors. Taken together, our data, for the first time, demonstrate the potential antitumor activity of DHA in HCC.     

新鬼臼毒素衍生物ZM-16诱导K562/A02细胞凋亡及其机制

鬼臼毒素衍生物是目前抗癌药物研究的特点,目前临床上应用较为广泛的依托泊苷(VP-16)和替尼泊苷(VM-26)均为半合成鬼臼毒素衍生物[1].     

Rottlerin exhibits antiangiogenic effects in vitro

Rottlerin, a natural product purified from Mallotus philippinensis, has a number of target molecules and biological effects. We recently found that Rottlerin caused growth arrest in MCF-7 breast cancer cells and human immortalized keratinocytes, through inhibition of NFκB and downregulation of cyclin D-1. To evaluate whether this effect could be generalized to primary cells, human microvascular endothelial cells were treated with Rottlerin. In this study, we demonstrated that Rottlerin prevents basal and TNFα-stimulated NFκB nuclear migration and DNA binding also in human microvascular endothelial cell, where NFκB inhibition was accompanied by the downregulation of NFκB target gene products, such as cyclin D-1 and endothelin-1, which are essential molecules for endothelial cell proliferation and survival. Rottlerin, indeed, inhibited human microvascular endothelial cells proliferation and tube formation on Matrigel. Rottlerin also increases cytoplasmic free calcium and nitric oxide levels and downregulates endothelin converting enzyme-1 expression, thus contributing to the drop in endothelin-1 and growth arrest. These results suggest that Rottlerin may prove useful in the development of therapeutic agents against angiogenesis.     

Analogs,formulations and derivatives of imatinib: a patent review

Introduction: The Bcr-Abl inhibitor imatinib was approved in 2001 for chronic myeloid leukemia therapy, and dramatically changed the lives of patients affected by this disease. Since it also inhibits platelet derived growth factor receptor (PDGFR) and c-Kit, imatinib is used for various other tumors caused by abnormalities of one or both these two enzymes.

Areas covered: This review presents an overview on imatinib formulations and derivatives, synthetic methodologies and therapeutic uses that have appeared in the patent literature since 2008.

Expert opinion: Innovative imatinib formulations, such as nanoparticles containing the drug, will improve its bioavailability. Moreover, oral solutions or high imatinib content tablets or capsules will improve patient compliance. Some solid formulations and innovative syntheses that have appeared in the last few years will reduce the cost of the drug, offering big advantages for poor countries. Some recently patented efficacious imatinib derivatives are in preclinical studies and could enter clinical trials in the next few years. Overall, Bcr-Abl inhibitors constitute a very appealing research field that can be expected to expand further.     

伊马替尼治疗Ph阳性进展期慢性粒细胞白血病

目的:观察伊马替尼治疗Ph阳性进展期慢性粒细胞白血病(CML)的疗效和耐药情况,研究改善伊马替尼耐药的方法。方法:32例Ph阳性进展期CML病人,其中加速期12例,急变期20例,每日口服伊马替尼600或800mg,持续3～9mo。结果:CML加速期病人血液学完全缓解率和总有效率分别为42%和83%,主要细胞遗传学缓解率25%,持续完全血液学缓解病例占25%。CML急变期各类型病人血液学完全缓解率和总有效率分别为20%和55%,主要细胞遗传学缓解率15%,持续完全血液学缓解病例占10%。CML急变期原发耐药和继发耐药分别为45%和20%,联合化疗与暂停伊马替尼对继发耐药可暂时改善其耐药性,但药物有效时间明显缩短。结论:伊马替尼对初治或复治的CML加速期和急变期病人均有效,可作为非移植CML治疗的标准一线方案,伊马替尼治疗CML急变期的原发耐药和继发耐药率较高,联合化疗和暂停伊马替尼可暂时改善其耐药性。     

Assessing the impact of substandard copy medicines in developing countries: the experience with imatinib copies

Access to safe, high-quality medicines is an important international healthcare issue. In developing countries, copy medicines may be attractive due to low acquisition price, but their potential risks due to inadequately demonstrated bioequivalence have garnered only limited attention. As a result, policy-makers, physicians and patients may have incomplete information regarding their real-world safety and efficacy. Using chronic myeloid leukemia (CML) treatment as an example, we conducted a literature review of case reports and study publications to assess whether the available literature provides evidence on the real-world safety and efficacy of imatinib copies. While several publications described clinical outcomes with imatinib copy treatment, significant gaps in interpretability and quality exist. We conclude that clear demonstration of bioequivalence is critical for copy drugs, and in the presence of uncertain bioequivalence, greater pharmacovigilance and real-world data benchmarked against originator medications are needed to assess the impact of copy medicines and protect patients in developing countries.     

大剂量伊马替尼、达沙替尼和尼洛替尼治疗慢性髓性白血病的成本效用分析

目的：分析大剂量伊马替尼、达沙替尼和尼洛替尼治疗对标准剂量伊马替尼耐药的慢性髓性白血病患者的成本效用。方法：计算三种治疗方案下患者的效用和花费的成本,在马尔可夫模型（Markov）中以3个月为周期进行5年的模拟并对结果做成本效用比较。结果：尼洛替尼治疗方案累计成本为1 595 289.10元,健康效用为1.276 4质量调整生命年（QALYs）,相比于尼洛替尼,大剂量伊马替尼组的增量成本效用比（ICUR）为-22 759 433.08（￥/QALYs）,达沙替尼组为-30 960 469.51（￥/QALYs）。结论：尼洛替尼方案累计成本最低且获得最多的QALYs,为绝对优势方案。     

The potentiation of menadione on imatinib by downregulation of ABCB1 expression

Imatinib was the first BCR-ABL inhibitor used in clinical practice to treat chronic myeloid leukaemia (CML) and significantly improve the life expectancy of CML patients in the chronic phase. However, a portion of CML patients are resistant to imatinib. This study aimed to determine whether menadione (Vitamin K3) can improve imatinib efficacy in CML and to thoroughly explore the combination regimen mechanism between imatinib and menadione. Menadione improved imatinib efficacy in K562 cells by downregulating ABCB1 expression and increased the intracellular concentration of imatinib, which confirmed that this combination regimen is more effective than imatinib monotherapy. The results demonstrate that menadione and imatinib combination therapy may be a promising approach to refractory CML.