ABSTRACT
Chronic lymphocytic leukemia (CLL) is a clonal malignant disease of B lymphocytes (T lymphocytes are rare) and usually occurs in elderly people. CLL has a highly variable clinical course, with a median survival of 35 to 63 months. In the era of immunochemotherapy, the survival of CLL patients has improved significantly, but most patients still have primary drug resistance and relapse after the treatment. The emergence of Bruton tyrosine kinase inhibitor has completely changed the treatment mode of CLL, making the treatment of CLL into the era of targeted therapy. Ibrutinib combined with CD19 chimeric antigen receptor T-cell has good efficacy and can improve the prognosis of patients.
ABSTRACT
OBJECTIVE To establish a method to detect the blood concentration of ibrutinib and apply it to the clinic. METHODS Using zanubrutinib as internal standard， the concentration of ibrutinib was detected by high performance liquid chromatography （HPLC） after plasma samples were processed by solid-phase extraction. The separation was performed on an Agilent 5 TC-C18（2） column with acetonitrile-0.5% potassium dihydrogen phosphate solution （43∶57， V/V） as the mobile phase at a flow rate of 1 mL/min， a detection wavelength of 260 nm， a column temperature of 40 ℃ ， a sample size of 20 μL， and a run time of 25 min. The concentration of ibrutinib was measured in the plasma of 9 patients with non-Hodgkin’s lymphoma 2 h after drug administration on the 30th day by the above method. RESULTS The linear range of the assayed mass concentration of ibrutinib was 10-500 ng/mL （R 2＝0.998 9）， the lower limit of quantification was 10 ng/mL， and the RSDs of the intra-batch and inter-batch precision tests were not higher than 12.77%. The recoveries of the extraction were 74.80% and 97.70%， with both RSDs＜2.90%， and the RSDs of the stability tests were not higher than 7.10%. The peak plasma concentrations of 9 patients were 15.341-279.628 ng/mL. CONCLUSIONS The established HPLC method is simple and rapid， and can be used for the determination of ibrutinib concentration in plasma samples.
ABSTRACT
Interleukin-1 receptor-associated kinase 4 (IRAK4) is a pivotal enzyme in the Toll-like receptor (TLR)/MYD88 dependent signaling pathway, which is highly activated in rheumatoid arthritis tissues and activated B cell-like diffuse large B-cell lymphoma (ABC-DLBCL). Inflammatory responses followed by IRAK4 activation promote B-cell proliferation and aggressiveness of lymphoma. Moreover, proviral integration site for Moloney murine leukemia virus 1 (PIM1) functions as an anti-apoptotic kinase in propagation of ABC-DLBCL with ibrutinib resistance. We developed a dual IRAK4/PIM1 inhibitor KIC-0101 that potently suppresses the NF-κB pathway and proinflammatory cytokine induction in vitro and in vivo. In rheumatoid arthritis mouse models, treatment with KIC-0101 significantly ameliorated cartilage damage and inflammation. KIC-0101 inhibited the nuclear translocation of NF-κB and activation of JAK/STAT pathway in ABC-DLBCLs. In addition, KIC-0101 exhibited an anti-tumor effect on ibrutinib-resistant cells by synergistic dual suppression of TLR/MYD88-mediated NF-κB pathway and PIM1 kinase. Our results suggest that KIC-0101 is a promising drug candidate for autoimmune diseases and ibrutinib-resistant B-cell lymphomas.
ABSTRACT
Objective:To investigate the clinical characteristics of steroid-resistant chronic graft-versus-host disease (cGVHD) patients and the therapeutic effect of ibrutinib.Methods:The clinical data of 3 steroid-resistant cGVHD patients treated with ibrutinib after allogeneic hematopoietic stem cell transplantation in the First Affiliated Hospital of Soochow University from December 2017 to March 2018 were retrospectively analyzed, and the literature was reviewed.Results:All 3 patients had different degrees of skin and oral cGVHD. One patient's oral symptom improved after the application of prednisone, and the skin symptom was better after oral ibrutinib; one patient's oral symptom improved after oral ibrutinib, but skin symptom did not improve significantly; one patient's skin symptom did not improve significantly. None of the 3 patients presented with adverse reactions such as hemorrhage, infection and cytopenia.Conclusions:Ibrutinib has a certain effect on the improvement of symptoms in steroid-resistant cGVHD patients.
ABSTRACT
Hemophagocytic syndrome (HPS) is a rare, life-threatening inflammatory response syndrome characterized by overactivation of the immune system, which leads to organ damage. Secondary HPS is usually triggered by infection, tumor and autoimmune disease. It has been clinically found that many HPS-like manifestations also occur during drug therapy. This article reviews the related progress of HPS induced by immune checkpoint inhibitors, ibrutinib and lamotrigine, in order to provide a guidance for clinical practice.
ABSTRACT
Objective:To investigate the clinical efficacy and safety of ibrutinib in treatment of chronic lymphocytic leukemia (CLL).Methods:The clinical data of 68 patients with CLL in Fujian Medical University Union Hospital from May 1998 to October 2019 were retrospectively analyzed, including 39 cases receiving ibrutinib as the first therapy, 20 cases receiving ibrutinib as the second therapy, and 9 cases receiving ibrutinib as the second and above therapy. The clinical characteristics, IGHV gene mutation, the short-term therapeutic efficacy and survival of patients with stratified chromosomal karyotype were analyzed; the adverse events were also analyzed.Results:The median follow-up time was 53.2 months until May 2020. The objective remission rate (ORR) was 83.8% (57/68), including 8 cases (11.8%) of complete remission, 49 cases (72.1%) of partial remission, 5 cases (7.4%) of the stable disease, 6 cases (8.8%) of progression of the disease. The ORR of patients without IGHVmutation was higher than that of those with IGHV mutation [93.3% (28/30) vs. 76.3% (29/38), χ2=33.656, P<0.05]; the ORR of patients with low risk and low-moderate risk International Prognostic Index (IPI) score was higher than that of those with moderate-high risk and high risk [90.6% (29/32) vs. 77.8% (28/36), χ2=7.248, P = 0.007], and differences in the ORR of patients stratified by other factors were not statistically significant (all P > 0.05). Among 68 patients, 31 cases (45.6%) had adverse reactions and they insistently received the treatment; 26 cases (38.2%) had grade1-2 adverse reactions, 5 cases (7.4%) had grade 3 and above adverse reactions; 2 cases (2.9%) had drug withdrawal because of adverse reactions. The median progression-free survival (PFS) time and overall survival (OS) time of CLL patients treated with ibrutinib had not been reached. The 5-year PFS rate of patients with IGHV mutation was higher than that of patients with IGHV non-mutation (100.0% vs.72.1%, P = 0.020), the 5-year PFS rate of patients with normal chromosome karyotype was higher than that of those with abnormal chromosome karyotype (100.0% vs.74.3%, P = 0.019). Conclusion:Ibrutinib is effective and safe in treatment of CLL patients.
ABSTRACT
When chimeric antigen receptor T cells (CAR-T) failed to treat relapsed and refractory B-cell malignancies, some researchers try to improve the efficacy by enhancing the function of CAR-T. It is a new hotspot of drug development and clinical research, and significant achievements have been made in this area recently. Multi-targeted CAR-T, lenalidomide, decitabine, programmed death 1 inhibitor and ibrutinib can all enhance the function of CAR-T. This article reviews the recent progress of enhancing the function of CAR-T in relapsed and refractory B-cell malignancies.
ABSTRACT
The aim of this paper is to analyze the global patent layout of ibrutinib, the patent layout of key themes, and the original research company's patent layout of ibrutinib, so as to provide a reference for the related domestic drug companies to be acquainted with the patent status of the drug, and to select and decide their breakthrough points and market competition strategies. In this study, the drug name and drug structure were used as keywords for search in the patent databases CNABS and DWPI, and the results were combined with the search results from the International Online Search System STN. Then, the patent with ibrutinib as the invention point was selected and used for further analysis. Pharmacyclics has applied for 26 patents on ibrutinib in China, and the subject matter relates to compounds, compositions, new uses, dosage forms, preparation methods and so on. Among them, 4 patents have been granted, 8 patents have been rejected or overdue, and 14 patent applications are still pending. In order to break through the patent barriers of ibrutinib, Chinese pharmaceutical companies should always pay much attention to the patent approval process and the expiration status of patents from the original research company, and make full use of the intellectual property system to effectively protect their own innovations.
ABSTRACT
Objective: The main aim of the research was to develop a fast and highly sensitive bioanalytical LC-MS/MS technique for the quantitation of ibrutinib in human plasma. Methods: Chromatography has achieved on a reverse phase-symmetry C18 (75 mm × 4.6 mm, 3.5 µm) column with gradient elution by acetonitrile, methanol and 0.1%v/v formic acid as the mobile phase. Chromatographic peaks were resolved with 0.7 ml/min flow rate. Drug was extracted with ethyl acetate solvent by liquid-liquid extraction method. Monitoring of transition of m/z 441.2 and 55.01 for ibrutinib and 446.5 and 60.01 for Ibrutinib-D5 were made on multiple reaction monitoring. Results: Calibration curve of ibrutinib was linear over 1-600 ng/ml concentration range with a regression coefficient (r2) value of>0.99. The % RSD values were less than 8.5% for inter-day and intra-day precision and accuracy. The method has excellent recovery and the percentage recovery values of lower quality control (LQC), median quality control (MQC) and higher quality control (HQC) samples were 101.86%, 102.8%, and 99.28% respectively. Conclusion: The drug was stable for more time at variable stability conditions and method was successfully applicable to the regular analysis of ibrutinib in biological matrices.
ABSTRACT
Chronic lymphocytic leukemia(CLL) is a heterogeneous mature B lymphocytic tumor.The apoptosis of mature lymphocyte is inhibited and clonal proliferation of mature lymphocyte aggregates in blood, bone marrow, spleen and lymph nodes, resulting in a class of inert hematological tumors.At present, the clinical pathogenesis is not completely clear, environmental and occupational factors do not occupy a major position.Studies have shown that long-term exposure to low-frequency electromagnetic fields may be associated with its incidence, but patients with primary and secondary relatives of lymphatic malignancies increased incidence.Many families still have patients whose age is earlier and the disease is more serious.In recent years, with the continuous improvement of medical level, a variety of treatment methods for chronic lymphoblastic leukemia have emerged, including a variety of new drugs.Ibutinib is the world's first marketed Bruton's tyrosine kinase(BTK) inhibitor, for more patients with chronic lymphoblastic leukemia has brought the gospel.This article reviews the clinical research progress of ibrutinib in treating CLL.
ABSTRACT
Objective@#To improve the knowledge and experience of ibrutinib combined with CAR-T cells in the treatment of high-risk chronic lymphoblastic leukemia (CLL) patients or small lymphocytic lymphoma (SLL) with TP53 gene aberration.@*Methods@#One case of del (17p) CLL patients with BCL-2 inhibitor resistance was treated with ibrutinib combined with CAR-T cells, successfully bridged to allogeneic hematopoietic stem cell transplantation (allo-HSCT) , and the relative literatures were reviewed.@*Results@#The patient was a young female with superficial lymph node enlarging at the beginning of the onset. Lymph node biopsy was confirmed as small lymphocytic lymphoma (SLL) without del (17p) . The disease progressed rapidly to CLL/SLL with del (17p) and bone marrow hematopoietic failure 2 years later. Firstly, the patient was treated with BCL-2 inhibitor (Venetoclax) , and the enlarged lymph nodes shrank significantly 2 months later. After 3 months, the disease progressed rapidly. The spleen was enlarged to 16 cm below the ribs, the neck lymph nodes was rapidly enlarged, and the superior vena cava syndrome appeared, which were mainly attributed to venetoclax resistance; so BTK inhibitor (ibrutinib) was used continuously after venetoclax discontinuation. Partial remission (PR) was achieved without lymphocytosis after 2 months, then ibrutinib was combined with CAR-T cells targeting CD19 antigen. Grade 1 of cytokine release syndrome (CRS) appeared after CAR-T cells infusion, and the complete remission (CR) was achieved after 1 month both in bone marrow and peripheral blood, with minimal residual disease (MRD) negative, then bridging allo-HSCT after 2 months of combined therapy.@*Conclusion@#CLL/SLL patients with TP53 aberration have poor prognosis because of rapid progression, drug resistance, etc. Ibrutinib combined with CAR-T cell therapy can quickly achieved complete remission.
ABSTRACT
As a first generation Bruton's tyrosine kinase inhibitor, ibrutinib is an oral small molecule targeted drug that has been prov-en to exhibit high efficacy and good safety in the treatment of B-cell lymphomas, including chronic lymphocytic leukemia or small lym-phocytic lymphoma (CLL/SLL) and mantle cell lymphoma (MCL). Since ibrutinib first became available in China, increasing numbers of Chinese patients have benefited from it; however, with the increase in the number of patients, clinicians are faced with more adverse events, such as bleeding, atrial fibrillation, diarrhea, joint pain, among others, than traditional chemotherapeutic drugs. Because of the unique mechanism of action of ibrutinib, it is also necessary to pay attention to drug interactions when dealing with these adverse events. This review comprehensively summarizes the treatment strategies for ibrutinib adverse events in the literature and hopes to provide a reference for clinicians.
ABSTRACT
OBJECTIVE： To establish a method for the determination of ibrutinib concentration in Beagle dogs， and to compare the pharmacokinetic difference of ibrutinib and its phospholipid complex in Beagle dogs. METHODS： The male Beagle dogs were randomly divided into Ibrutinib suspension group and Ibrutinib phospholipid complex group （using 0.5％ Carboxymethylcellulose sodium solution and water as solvent， mass concentration of 5 mg/mL）， with 3 dogs in each group. All Beagle dogs were given relevant medicine suspension （15 mg/kg） intragastrically， and 2 mL of blood were collected from the forelimb vein before administration and 0.017， 0.083， 0.25， 0.5， 1， 2， 4， 8 and 12 h after administration. Plasma concentration of ibrutinib was were determined by HPLC. Using tolbutamide as internal standard， the determination was performed on Betasil C18 column with mobile phase consisted of acetonitrile-water （contained 0.5％ triethylamine， pH value adjusted to 3.2 with glacial acetic acid）（45 ∶ 55， V/V） at the flow rate of 1.0 mL/min. The detection wavelength was set at 256 nm， and the column temperature was 25 ℃. The sample size was 20 μL. The pharmacokinetic parameters of Beagle dogs in 2 groups were calculated by using DAS 2.1.1 software. The difference of ibrutinib and its phospholipid complex were investigated by t-test. RESULTS： The linear range of ibrutinib was 5-5 000 ng/mL （r＝0.999 8）； lower limit of quantitation was 5 ng/mL； minimum detection limit was 1.3 ng/mL. RSDs of intra-batch and inter-batch were lower than 10％； the accuracy was 98.81％-106.20％； the extraction method did not influence the determination of the substance to be measured. Pharmacokinetic parameters of Ibrutinib suspension and Ibrutinib phospholipid complex with signal intragastric administration were as follows： tmax were（2.00±0.09） and （0.25±0.03）h； cmax were（610.67±21.36） and （2 308.72±100.41）ng/mL； AUC0-12 h were （4 516.67±383.43） and （9 394.16±874.21）ng·h/mL； AUC0-∞ were （6 174.32±525.27） and （10 717.33±897.62）ng·h/mL，with statistical significance （P＜0.05）. The relative bioavailability of Ibrutinib phospholipid complex was 207.99％. CONCLUSIONS： Established HPLC method is simple， specific and sensitive， and can be used for plasma concentration determination and pharmacokinetic study of ibrutinib. The pharmacokinetic parameters of phospholipid complex prepared from ibrutinib changed significantly， drug absorption is accelerated and bioavailability is improved significantly.
ABSTRACT
OBJECTIVE： To observe the inhibitory effects and possible mechanism of new small molecular kinase inhibitors Ibr-7 [Irutinil（Ibr） derivatives] on human pancreatic cancer Capan-2 cells. METHODS： Taking Capan-2 cells as objects， CCK-8 method was used to determine the proliferation of cells after treated with 1， 2， 4， 8 μmol/L Ibr/Ibr-7 for 48 h. The survival rates of cells were calculated. Sensitization effects of 1 μmol/L Ibr/Ibr-7 on different doses of gemcitabine/paclitaxel （0.062 5， 0.125， 0.25， 0.5， 1 μmol/L） were detected. Clone formation test was used to detect the situation of cell clone formation after treated with 1， 2， 4 μmol/L Ibr/Ibr-7 for 48 h. The number of cell colony formation was recorded. Flow cytometry or JC-1 method was used to detect the apoptosis of cells after treated with 2， 4， 8 μmol/L Ibr-7 for 24 or 16 h and the changes of mitochondrial transmembrane potential； total apoptotic rate and the percentage of mitochondrial membrane potential decrease were calculated. Western blotting was used to detect the expression of related apoptotic protein （PARP， Noxa， Bcl-2， Bax， Mcl-1， Bcl-xL）. RESULTS： After treated with 1， 2， 4， 8 μmol/L Ibr/Ibr-7 for 48 h， the survival rates of cells were decreased significantly； those of Ibr-7 groups were significantly lower than those of same-dose Ibr groups； IC50 of Ibr-7 was significantly lower than that of Ibr （P＜0.05 or P＜0.01）. After combined with Ibr/Ibr-7， the survival rate of cells was significantly lower than that of same-dose gemcitabine/paclitaxel alone group， and the Ibr-7 combination group was significantly lower than same-dose Ibr combination group （P＜0.05 or P＜0.01）. After treated with 2， 4 μmol/L Ibr and 1， 2， 4 μmol/L Ibr-7 for 48 h， the number of cell clone formation was decreased significantly， while Ibr-7 groups were significantly lower than same-dose Ibr groups （P＜0.01）. After treated with different doses of Ibr-7 for 24 or 16 h， total apoptosis rate of cells （2， 4， 8 μmol/L）， the proportion of cell mitochondrial membrane potential decrease （8 μmol/L）， the relative protein expression of Noxa （2， 4， 8 μmol/L） and Bax （8 μmol/L） were increased significantly， while the protein expression of PARP （8 μmol/L）， Bcl-2 （4 μmol/L）， Mcl-1 （2， 4， 8 μmol/L） were decreased significantly； above indexes （except for relative expression of PARP and Bcl-2） of 8 μmol/L Ibr-7 group were significantly better than same-dose Ibr group （P＜0.05 or P＜0.01）. There was no statistical significance in protein expression of Bcl-xL among those groups （P＞0.05）. CONCLUSIONS： Compared with Ibr， Ibr-7 has better inhibitory and apoptotic effects on human pancreatic cancer Capan-2 cells in vitro， and has stronger chemotherapeutic drug sensitization activity， the mechanism of which may be associated with reducing mitochondrial transmembrane potential， down-regulating the protein expression of PARP， Bcl-2 and Mcl-1 and up-regulating the protein expression of Noxa and Bax.
ABSTRACT
Chronic lymphocytic leukemia(CLL) is a heterogeneous mature B lymphocytic tumor.The apoptosis of mature lymphocyte is inhibited and clonal proliferation of mature lymphocyte aggregates in blood,bone marrow,spleen and lymph nodes,resulting in a class of inert hematological tumors.At present,the clinical pathogenesis is not completely clear,environmental and occupational factors do not occupy a major position. Studies have shown that long -term exposure to low-frequency electromagnetic fields may be associated with its incidence,but patients with primary and secondary relatives of lymphatic malignancies increased incidence. Many families still have patients whose age is earlier and the disease is more serious.In recent years,with the continuous improvement of medical level,a variety of treatment methods for chronic lymphoblastic leukemia have emerged,including a variety of new drugs.Ibutinib is the world＇s first marketed Bruton＇s tyrosine kinase(BTK) inhibitor,for more patients with chronic lymphoblastic leukemia has brought the gospel.This article reviews the clinical research progress of ibrutinib in treating CLL.
ABSTRACT
As a novel molecular targeted anti﹣tumor drug, it has been proved that ibrutinib has remarkable anti﹣tumor activity and is characterized by high efficiency and low toxicity. However, the risks of drug resistance, safety, and disease transformation are gradually concerned. This review focused on the research progress of ibrutinib′s drug resistance mechanism in various B﹣cell non﹣Hodgkin lymphomas.
ABSTRACT
Objective: To develop an HPLC method for the determination of ibrutinib in rat's plasma, and study its pharmacoki-netics. Methods: The analytical column was packed with ZORBAX XDB-C18(150 mm×4. 6 mm,5 μm). A mixture of acetonitrile-wa-ter-0. 1% trifluoroacetic acid (42 ∶ 31 ∶ 27) was used as the mobile phase with the flow rate at 1. 0 ml·min-1. The detection wave-length was set at 258 nm. The column temperature was set at 30℃. Carbamazepine was used as the internal standard. Plasma was ex-tracted under alkaline condition and ibrutinib was detected. Nine SD rats were treated with single dose of ibrutinib 15 mg·kg-1by in-tragastric administration. Blood samples were collected at different time points after ibrutinib administration. The plasma concentration of ibrutinib was detected, and the pharmacokinetic parameters were calculated by DAS software. Results: Excellent linear relationship was obtained within the range of 10 μg·L-1to 2 000 μg·L-1(r =0.999 7). The intra-day RSDs were 7.11%, 10.41% and 3. 19% , and the inter-day RSDs were 2. 56% , 1. 98% and 3. 79% respectively for three concentrations ( 25, 500 and 1 500 μg· L-1), the average recoveries were (78. 91 ± 2. 10)% , (86. 29 ± 3. 97)% and (83. 61 ± 2. 11)% , respectively. After intragastric administration of ibrutinib, the main pharmacokinetic parameters of ibrutinib were as follows:Cmaxwas(1 019.43 ±74.85)μg·L-1, Tmaxwas(4.78 ±1.20)h, AUC(0-36)was(10 417.26 ±2 167.51)μg·h·L-1, AUC(0-inf)was(10 956.72 ±2491.09)μg·h·L-1, and t1/2was(8.57 ±1.47)h. Conclusion: The method is simple, rapid and accurate, and can be applied in the studies on the phar-macokinetics of ibrutinib.
ABSTRACT
OBJECTIVE To investigate the therapeutic effect and side effect of ibrutinib long-term treatment in systemic lupus erythematosus (SLE) model mice induced by pristane. METHODS Female 6-week old BALB/c mice were ip given pristane 0.5 mL once for SLE induction. Four weeks later, the SLE model mice were divided into three groups based on the body mass and serum level of anti-double-strand DNA (DS-DNA) antibody and treated with 1%methylcellulose (model control group, ig), ibrutinib (30 mg·kg-1, ig) or prednisone (10 mg·kg-1, ig), respectively, once daily for 28 weeks. Every 4 weeks, body mass of each mouse was measured. Autoantibodies against DS-DNA, single-strand DNA (SS-DAN), and histone were tested by ELISA. The incidence of lupus arthritis and clinical score of inflammation and edema were recorded and graded. Biochemical analysis of urea protein, serum urea nitrogen and creatinine was used to evaluate kidney function. Mice were euthanized post 28 weeks of dosing. Inter-leukin-6 (IL-6), interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) were tested by ELISA. The heart, liver, spleen, lung, and kidney were collected and weighed for organ relative mass calculation. Biochemical analysis of serum glutamic-pyruvic transaminase (GPT), glutamic-oxal(o)acetic transaminase (GOT) and alkaline phosphatase (ALP) was used to evaluate liver function. Hind feet were collected for HE staining and pathological scoring to observe renal injury and inflammation. Immunohistochemical staining was used to study IgG immune complexes deposition in kidneys of lupus. RESULTS Compared with normal control group, autoantibodies (anti-DS-DNA, anti-SS-DNA and anti-histone antibodies), renal function indexes (serum creatinine, urea nitrogen and urine protein), and cytokines (IL-6, IFN-γ and TNF-α) levels in model group were significantly increased (P<0.01). The model group mice had obvious clinical symptoms of arthritis (P<0.01), serious inflammation cell invasion (P<0.01), and the mass of kidneys and spleen increased significantly (P<0.01). Compared with model group, after 28 weeks of treatment, ibrutinib decreased the level of anti-DS-DNA, anti-SS-DNA and anti-histone antibodies (P<0.01), decreased the lupus arthritis score (P<0.01) and the morbidity of arthritis, reduced the level of cyto-kines IL-6, IFN-γand TNF-α(P<0.01), reduced the level of serum creatinine, serum urea nitrogen and urine protein (P<0.01), improved pathological symptoms of hind feet such as inflammation, cartilage destruction, bone resorption and pannus (P<0.01), alleviated renal tissue inflammatory cell invasion and the immune-complex precipitation, and reduced the mass of organs (spleen and kidneys, P<0.01) and the level of liver function (GPT and ALP, P<0.01). CONCLUSION Long-term treatment with ibrutinib has therapeutic effect on the model mice of SLE, and has no obvious side effect.
ABSTRACT
Objective:To detect Bruton tyrosin kinase (BTK) expression in patients with mantle cell lymphoma (MCL) and analyze its correlation with clinical features and prognosis. Methods:A total of 32 cases of MCL tissues and 10 cases of benign lymph nodes were sampled and stained with immunohistochemical (IHC) staining. Clinical data of these patients were analyzed using SPSS 17.0. Results:BTK was positively expressed in MCL and normal lymphoid tissues and was more strongly expressed in MCL tissue than in normal lym-phoid tissue. Moreover, BTK expression level was correlated with Ki-67 and MIPI scores. Prognosis analysis showed that patients with high BTK expression exhibited shorter progression-free survival (PFS) than patients with low expression levels (P=0.030);however, no significant difference in overall survival (OS) was observed (P=0.073). Single-factor analysis of PFS showed that age≥65 years, ECOG score≥2, bone marrow involvement, strongly positive BTK expression, Ki-67>30%, and MIPI score≥6 are poor prognostic factors for patients with MCL. Only MIPI score≥6 is considered an independent poor prognostic factor in the multivariate analysis. Conclusion:BTK is strongly and positively expressed in patients with MCL, and its expression level is correlated with Ki-67 and MIPI scores. Patients with high-level BTK expression usually exhibit shorter PFS than those with low-level BTK expression;however, owing to short follow-up time and limited sample size, high-level BTK expression cannot be considered an independent poor prognostic factor for PFS.
ABSTRACT
Objective:To illustrate the effect and mechanism of ibrutinib,a Bruton's tyrosine kinase(BTK)inhibitor that inhibits diffuse large B-cell lymphoma(DLBCL)cell survival.Methods:DLBCL cell lines SUDHL-10 and HBL-1 were treated with ibrutinib at different concentrations.A MTT assay was used to detect the inhibition of cell proliferation.Cell apoptosis was analyzed by Annexin V-binding assay,as well as flow cytometry and DAPI staining.The expression of phosphorylated BTK,AKT and ERK was detected by Western blot. DLBCL cells were co-cultured with MSC.The inhibitory effect of ibrutinib on DLBCL cells in tumor microenvironment was assessed in clonogenicity in vitro and in a tumor-bearing non-obese diabetic/severe combined immunodeficient mice in vivo.Results:Up to 2.5 μmol/L and high concentrations of ibrutinib significantly inhibited the proliferation of DLBCL cells in a dose-dependent manner.Approx-imately 1 and 2.5 μmol/L ibrutinib was added on SUDHL-10 cells for 24 h,and the cell apoptotic rates were(21.73±3.64)% and(34.71± 2.36)%,respectively.Both were superior to that of the control group(3.55±1.89)%(P<0.05).Both two DLBCL cell lines pretreated with 5 and 10 μmol/L ibrutinib for 24 h and exhibited nuclear shrinkage at 5 μmol/L and nuclear fragmentation at 10 μmol/L.The expres-sion of phosphorylated BTK,AKT,and ERK decreased significantly after ibrutinib treatment.Ibrutinib inhibited clonogenicity in vitro(P<0.01)and cell proliferation and growth in vivo of DLBCL cells in co-culture system.The differences were statistically significant.Conclu-sion:Ibrutinib can inhibit the proliferation and induce apoptosis of SUDHL-10 and HBL-1 cell lines through a mechanism of blocking the AKT and ERK signaling pathways,as well as the proliferation of DLBCL cells in tumor microenvironment.This finding can significant-ly benefit DLBCL treatment.