Mechanism of nuclear protein 1 in the resistance to axitinib in clear cell renal cell carcinoma / 北京大学学报(医学版)

OBJECTIVE@#To explore the potential mechanism of resistance to axitinib in clear cell renal cell carcinoma (ccRCC), with a view to expanding the understanding of axitinib resistance, facilitating the design of more specific treatment options, and improving the treatment effectiveness and survival prognosis of patients.@*METHODS@#By exploring the half maximum inhibitory concentration (IC50) of axitinib on ccRCC cell lines 786-O and Caki-1, cell lines resistant to axitinib were constructed by repeatedly stimulated with axitinib at this concentration for 30 cycles in vitro. Cell lines that were not treated by axitinib were sensitive cell lines. The phenotypic differences of cell proliferation and apoptosis levels between drug resistant and sensitive lines were tested. Genes that might be involved in the drug resistance process were screened from the differentially expressed genes that were co-upregulated in the two drug resistant lines by transcriptome sequencing. The expression level of the target gene in the drug resistant lines was verified by real-time quantitative polymerase chain reaction (RT-qPCR) and Western blot (WB). The expression differences of the target gene in ccRCC tumor tissues and adjacent tissues were analyzed in the Gene Expression Profiling Interactive Analysis (GEPIA) public database, and the impact of the target gene on the prognosis of ccRCC patients was analyzed in the Kaplan-Meier Plotter (K-M Plotter) database. After knocking down the target gene in the drug resistant lines using RNA interference by lentivirus vector, the phenotypic differences of the cell lines were tested again. WB was used to detect the levels of apoptosis-related proteins in the different treated cell lines to find molecular pathways that might lead to drug resistance.@*RESULTS@#Cell lines 786-O-R and Caki-1-R resistant to axitinib were successfully constructed in vitro, and their IC50 were significantly higher than those of the sensitive cell lines (10.99 μmol/L, P < 0.01; 11.96 μmol/L, P < 0.01, respectively). Cell counting kit-8 (CCK-8) assay, colony formation, and 5-ethynyl-2 '-deoxyuridine (EdU) assay showed that compared with the sensitive lines, the proliferative ability of the resistant lines decreased, but apoptosis staining showed a significant decrease in the level of cell apoptosis of the resistant lines (P < 0.01). Although resistant to axitinib, the resistant lines had no obvious new replicated cells in the environment of 20 μmol/L axitinib. Nuclear protein 1 (NUPR1) gene was screened by transcriptome sequencing, and its RNA (P < 0.0001) and protein expression levels significantly increased in the resistant lines. Database analysis showed that NUPR1 was significantly overexpressed in ccRCC tumor tissue (P < 0.05); the ccRCC patients with higher expression ofNUPR1had a worse survival prognosis (P < 0.001). Apoptosis staining results showed that knockdown ofNUPR1inhibited the anti-apoptotic ability of the resistant lines to axitinib (786-O, P < 0.01; Caki-1, P < 0.05). WB results showed that knocking downNUPR1decreased the protein level of B-cell lymphoma-2 (BCL2), increased the protein level of BCL2-associated X protein (BAX), decreased the protein level of pro-caspase3, and increased the level of cleaved-caspase3 in the resistant lines after being treated with axitinib.@*CONCLUSION@#ccRCC cell lines reduce apoptosis through theNUPR1 -BAX/ BCL2 -caspase3 pathway, which is involved in the process of resistance to axitinib.

ADE signal mining and analysis of axitinib based on FAERS database / 中国药房

OBJECTIVE To provide references for the clinical safe use of axitinib. METHODS Adverse drug event （ADE） data for axitinib were collected from the US FDA Adverse Event Reporting System （FAERS） database from the first quarter of 2012 to the fourth quarter of 2022. The data were mined and analyzed by utilizing the ratio-of-reporting-ratio （ROR） method and comprehensive standard method of the United Kingdom’s Medicines and Healthcare Products Regulatory Agency （MHRA） of proportional imbalance measurement. RESULTS A total of 13 962 reports of axitinib-related ADEs were obtained， with patients’ age concentrated in 65-85 years （43.25%）， gender predominantly male （65.23%）， country of reporting predominantly US （60.01%）， and serious ADE outcomes mostly hospitalization or prolonged hospitalization （31.51%）. A total of 172 ADE risk signals were detected， involving 18 system and organ classifications （SOC）， mainly systemic diseases and various reactions at the site of administration （3 749 cases， 30.84%） and gastrointestinal system diseases （2 067 cases， 17.00%）. ADE risk signals that occurred more frequently were generally consistent with the drug instruction， such as diarrhea， fatigue， and hypertension； new ADE risk signals requiring clinical attention were death， immune-mediated nephritis， and PT signals contained in the SOC of various benign， malignant， and tumors of undetermined nature （including cysts and polyps）. CONCLUSIONS For ADEs that occur frequently with axitinib and are already contained in the drug instruction （e.g. hypertension， diarrhea）， they should be adequately evaluated before administration， especially for patients with combined use of immune checkpoint inhibitors and patients with underlying hypertension； for ADEs with stronger signals and newer ADEs （e. g. death， disease progression， tumor progression）， the patient’s disease progression should be closely monitored during the treatment period for potentially fatal ADEs； for its rare ADEs （e. g.immune-mediated nephritis， scrotal ulcer， non-infectious encephalitis）， clinical validation should be further strengthened.

Immunotherapy-based first-line treatment of intermediate- and poor-risk advanced renal cell carcinoma: number needed to treat and cost of preventing an event from the perspective of the Brazilian private healthcare system / Tratamento baseado em imunoterapia para primeira linha do carcinoma renal avançado com risco intermediário ou alto: número necessário a tratar e custo para prevenir um evento na perspectiva do sistema privado de saúde brasileiro

Objective: To perform an analysis over time of the number needed to treat (NNT) and the cost of preventing an event (COPE) for nivolumab + ipilimumab (NIVO+IPI) and pembrolizumab + axitinib (PEMBRO+AXI) as first-line treatments for advanced renal cell carcinoma patients with intermediate or poor-risk, under the Brazilian private healthcare system perspective. Methods: The NNT for overall survival (OS) and progression-free survival (PFS) from 12-month to maximum available follow-up from CheckMate 214 and KEYNOTE-426 studies were used to estimate the COPE. Treatment costs were estimated considering the labeled dosing and median PFS as a proxy for treatment duration. Results: The OS NNT for NIVO+IPI decreased from 12 to 8 and for PEMBRO+AXI increased slightly from 7 to 8 at 12 and 42 months, respectively. For PFS, NNT for NIVO+IPI decreased from 15 to 6, and for PEMBRO+AXI increased from 7 to 10 at 12 and 30 months. The estimated treatment cost is R$ 638,620 for an estimated median of 11.2 months of NIVO+IPI treatment and R$ 966,818 for 13.8 months of PEMBRO+AXI treatment. COPE for OS at 12 and 42 months was R$ 7,663,440 and R$ 5,108,960 with NIVO+IPI and R$ 6,047,417 and R$ 7,734,547 with PEMBRO+AXI. For PFS, COPE at 12 and 30 months was R$ 9,579,300 and R$ 3,831,720 with NIVO+IPI and R$ 6,047,417 and R$ 9,668,184 with PEMBRO+AXI. Conclusions: Treatment with NIVO+IPI results in lower COPE than PEMBRO+AXI from month 18 onwards, driven by lower treatment costs and improved NNT over time with NIVO+IPI

Objetivo: Analisar ao longo do tempo o número necessário a tratar (NNT) e o custo para prevenir um evento (COPE) para nivolumabe + ipilimumabe (NIVO+IPI) e pembrolizumabe + axitinibe (PEMBRO+AXI) na primeira linha de tratamento do carcinoma de células renais avançado com risco intermediário ou alto na perspectiva do sistema suplementar de saúde brasileiro. Métodos: O NNT para sobrevida global (SG) e sobrevida livre de progressão (SLP) para 12 meses até o máximo de tempo de seguimento disponível dos estudos CheckMate 214 e KEYNOTE-426 foi usado para estimar o COPE. Custos de tratamento foram estimados considerando a dosagem em bula e a mediana de SLP como aproximação para duração de tratamento. Resultados: O NNT de SG para NIVO+IPI reduziu de 12 para 8 e para PEMBRO+AXI subiu de 7 para 8 em 12 e 42 meses, respectivamente. Para SLP, NIVO+IPI teve redução de 15 para 6 e para PEMBRO+AXI aumentou de 7 para 10 em 12 e 30 meses. O custo estimado é de R$ 638.620 para mediana de 11,2 meses de tratamento com NIVO+IPI e de R$ 966.818 para 13,8 meses com PEMBRO+AXI. O COPE para SG foi de R$ 7.663.440 e R$ 5.108.960 com NIVO+IPI e de R$ 6.047.417 e R$ 7.734.547 com PEMBRO+AXI para 12 e 42 meses. Para SLP, foi de R$ 9.579.300 e R$ 3.831.720 com NIVO+IPI e de R$ 6.047.417 e R$ 9.668.184 com PEMBRO+AXI em 12 e 30 meses. Conclusões: O tratamento com NIVO+IPI resulta em menor COPE, em comparação com PEMBRO+AXI, a partir de 18 meses de seguimento, justificado por menor custo de tratamento e melhora do NNT ao longo do tempo com NIVO+IPI

Effects of multi-target tyrosine kinase inhibitor Acitinib on liver fibrosis / 西安交通大学学报(医学版)

【Objective】 To observe the therapeutic effects of Axitinib, a tyrosine kinase receptor inhibitor, on liver fibrosis. 【Methods】 In vivo, CCL4 was injected intraperitoneally to induce liver fibrosis in mice. After modeling, Axitinib-carboxymethyl cellulose (Axitinib-CMC) solution or CMC solution were administered by gavage. After 2 weeks of modeling, 4 weeks of modeling, 1 week of treatment and 2 weeks of treatment, the mice were killed and their liver tissues were stained by HE, Masson and α - SMA immunohistochemistry. In vitro, human hepatic stellate cell line (LX2) and human normal liver cell line (LO2) were intervened with different concentrations of Axitinib-CMC solution. MTT assay was performed 48 h and 72 h after the intervention. Flow cytometry was used to observe the apoptosis of the two cell lines. Western blotting was used to detect the expressions of Fas, Caspase-8, Caspase-3 and Bcl-2 proteins. 【Results】 HE and Masson staining results showed that CCL4 could induce liver fibrosis in the mice, and the degree of liver fibrosis was more severe in the 4-week than that in the 2-week treatment group. After treatment with Axitinib, the collagen staining area and the positive expression level of α-SMA were significantly lower than those in 4-week group and CMC treatment control group (P<0.05). In vitro, Axitinib could effectively inhibit the viability of hepatic stellate cell line LX2 and promote its apoptosis. Meanwhile, the expressions of pro-apoptotic proteins Fas, Caspase-8 and Caspase-3 increased, while the expression of apoptosis suppressor gene Bcl-2 decreased. However, the above changes were not found in control hepatocyte line LO2. 【Conclusion】 Axitinib exerts an anti-fibrosis effect by inducing apoptosis of hepatic stellate cells, and has no significant effect on normal hepatocytes.

Combination of axitinib with dasatinib improves the outcome of a chronic myeloid leukemia patient with BCR-ABL1 T315I mutation / 中南大学学报(医学版)

Chronic myeloid leukemia (CML) is one of the most common hematological malignancies and characterized by the formation of Philadelphia (Ph) chromosome. Recently, tyrosine kinase inhibitors (TKI) treatment greatly improved the prognosis of CML. However, the options may be limited when a patient develops traditional TKI resistance or gene mutation. Herein, we reported a case. A 38-year-old male CML patient developed a BCR-ABL1 gene mutation of T315I after 2.5 years of TKI treatment, including imatinib and dasatinib. We adjusted the treatment with the combined application of dasatinib and axitinib. BCR-ABL1 gene copies dropped down and achieved an early molecular response at 2 months later. Subsequently, he received hematopoietic stem cell transplantation. Axitinib and dasatinib were applied for another half year after the allogeneic hematopoietic stem cell transplantation (allo-HSCT). Two years after the allo-HSCT, the BCR-ABL1 gene was still undetectable. It provided a successful example in treating CML patients carrying BCR-ABL1 T315I mutation via combination of axitinib with conditional TKI.

Efficacy of axitinib plus sintilimab in intermediate-and high-risk advanced renal cell carcinoma / 中国肿瘤临床

Objective: To investigate the preliminary efficacy and safety of axitinib plus sintilimab in the treatment of intermediate- and high-risk advanced renal cell carcinoma. Methods: A retrospective study of patients with advanced renal cell carcinoma treated with axitinib and sintilimab was conducted in Peking University Cancer Hospital & Institute between April 2019 to December 2019. There were seven cases of clear-cell renal cell carcinoma and three cases of non-clear-cell renal cell carcinoma.All patients received 200 mg sintilimab intravenously every 3 weeks and 5 mg axitinib orally twice daily. Objective response rate (ORR), progression-free survival (PFS), and adverse effects were analyzed. Result: With the median age of 58.5 years (range 43.0-67.0), the patients were all classified as intermediate- and high-risk patients, according to the international metastatic renal cell carcinoma database consortium (IMDC) criteria. The overall ORR was 40.0% (4/10), and the disease control rate (DCR) was 90.0% (9/10). The ORR was 57.1% (4/7) in patients with clear-cell renal cell carcinoma. The main adverse effects included elevation of hepatic transaminases (number of patients, n=4; 40.0%), hypothyroidism (n=4; 40.0%), nausea (n=3; 30.0%), hypertension (n=2; 20.0%), and hand-foot skin reaction (n=2; 20.0%). Most adverse effects were of grade 1 or 2, but grade 3-4 adverse events occurred in three patients.Most adverse effects were ameliorated by effective symptomatic treatment. Conclusions: Axitinib plus sintilimab achieved promising ORR and DCR in patients with intermediate- and high-risk advanced renal cell carcinoma, with tolerable adverse effects.

Tumor renal benigno versus carcinoma renal: reporte de caso y revisión de la literatura / Benign Kidney Tumor versus Renal Carcinoma: Case Report and Literature Review

Introducción El cáncer renal representa 2,4% de los casos diagnosticados de cáncer en la población general, es más común en hombres que en mujeres, y se presenta con más frecuencia entre la 6ta y la 8ta décadas de vida. Se estima que el 16% de los pacientes se diagnostican como enfermedad metastásica. Objetivo Se presenta el caso de un paciente cuyo diagnóstico de carcinoma renal se confundió inicialmente con un tumor benigno. Métodos A un hombre de 56 años de edad se le realizó hace 3 años ese diagnóstico en un estadio avanzado de la enfermedad, a pesar del hallazgo incidental de una masa, que se consideró benigna durante 5 años. Resultados Al momento del diagnóstico de carcinoma de células claras, el tumor era Estadio IV, con metástasis a pulmón. Recibió primera línea de tratamiento con sunitinib, pero fue suspendido por toxicidad; segunda línea con pazopanib durante 1 año, después presentó progresión de la enfermedad, por lo cual se cambió a tratamiento con axitinib con respuesta parcial, sin embargo, se suspendió por toxicidad cardiaca, entre otras. Al momento el paciente ha recibido 5 ciclos de bevacizumab con adecuada tolerancia. Conclusiones Es necesario resaltar la indicación de diagnóstico adecuado y manejo quirúrgico en masas renales sospechosas.

Introduction Kidney cancer represents 2.4% of diagnosed cases of cancer in the general population; it is more common in men than in women, and occurs more frequently between the 6th and 8th decades of life. It is estimated that 16% of patients are diagnosed as metastatic disease. Objective To report the case of a male patient whose diagnosis of renal carcinoma was initially misdiagnosed as a benign tumor. Methods We present a 56-year-old male diagnosed three years back with malignancy at an advanced stage of the disease, despite the incidental finding of a tumor that for 5 years was considered benign. Results At the time of diagnosis of clear cell carcinoma, the tumor was Stage IV, with lung metastasis. He received first line treatment with sunitinib, which was discontinued due to toxicity. Subsequently, a second line with pazotinib for 1 year, then presented progression of the disease, so treatment was changed to axitinib with partial response., It was discontinued, however, due to cardiac toxicity, among others. At the time of writing, the patient has received 5 cycles of bevacizumab with adequate tolerance. Conclusions It is necessary to highlight the need for adequate diagnosis and surgical management in suspicious renal masses.

Effect of axitinib on the proliferation and apoptosis of human lung adenocarcinoma PC9 cells and its mechanism / 肿瘤研究与临床

Objective To study the effect of axitinib on the proliferation and apoptosis of human lung adenocarcinoma PC9 cells and its mechanism. Methods PC90 cells were treated with different concentrations (0, 0.125, 0.25, 0.5, 1, 2, 4, 8, 16, 32, and 64 μmol/L) of axitinib for 72 h, and half-inhibitory concentration (IC50) was calculated. The cell proliferation ability was detected by CCK-8 method. Plate cloning experiments were performed to observe the effect of axitinib on the formation of PC9 cell clones. The mitochondrial membrane potential and apoptosis of PC9 cells were detected by flow cytometry. The expression of cleaved-Caspase-3 protein in PC9 cells was detected by Western blot. Results Asitinib inhibited the proliferation of PC9 cells in a concentration-dependent manner. The IC50 at 72 h was 10.18μmol/L. The clone formation rates of PC9 cells were (100.0±3.2)％, (58.6±2.7)％, (29.3±3.3)％, and (10.9±3.0)％10 d after treatment with 0, 1, 2 and 4 μmol/L axitinib, and the difference was statistically significant (F= 316.922, P< 0.01). The apoptotic rate of PC9 cells at early and late stages increased after treatment with different concentrations of axitinib for 48 h, and the differences were statistically significant (both P< 0.01). After treatment with 0, 4, 8 and 16 μmol/L axitinib for 24 h, the percentage of PC9 cells with low mitochondrial membrane potential was (11.9±1.9)％, (38.5±2.3)％, (56.3±2.7)％, and (76.9±3.1)％, and the difference was statistically significant (F=234.320, P<0.01). The expression level of cleaved-Caspase-3 protein in PC9 cells increased, and the relative expression levels were 1.00±0.04, 1.26±0.09, 1.78±0.12, and 2.10±0.11, respectively, and the difference was statistically significant (F=55.670, P<0.01). Conclusions Axitinib could inhibit the proliferation of human lung adenocarcinoma PC9 cells. Axitinib induces the apoptosis of PC9 cells possibly through decreasing the mitochondrial membrane potential of PC9 cells.

HPLC method development and validation for the estimation of Axitinib in rabbit plasma

ABSTRACT A rapid, sensitive, and accurate high performance liquid chromatography for the determination of Axitinib (AN) in rabbit plasma is developed using crizotinibe as an internal standard (IS). Axitinib is a tyrosine kinase inhibitor, used in the treatment of advanced kidney cancer, which works by slowing or stopping the growth of cancer cells. The chromatographic separation was performed on a Waters 2695, Kromosil (150 mm × 4.6 mm, 5 µm) column using a mobile phase containing buffer (pH 4.6) and acetonitrile in the ratio of 65:35 v/v with a flow rate of1 mL/min. The analyte and internal standard were extracted using liquid-liquid extraction with acetonitrile. The elution was detected by photo diode array detector at 320 nm.The total chromatographic runtime is 10.0 min with a retention time for Axitinib and IS of 5.685, and 3.606 min, respectively. The method was validated over a dynamic linear range of 0.002-0.2µg/mL for Axitinib with a correlation coefficient of r2 0.999.

Synergistic anti-tumor effects of axitinib and doxorubicin / 药学学报

To study the synergistic anti-tumor effects of doxorubicin and axitinib in combination, and investigate the underlying mechanism, we performed the MTT cytotoxic assay and tumor spheroids inhibition to investigate in vitro using A549 cells. The cell internalization, cell cycle distribution and DNA ladder experiments were performed to study the synergistic mechanisms. A549 xenograft was established in nude mice and adopted to study the in vivo anti-tumor effect of doxorubicin and axitinib. Results showed that combination of doxorubicin and axitinib exerted significantly higher cytotoxicity than each single drug, and induced a synergistic effect on tumor spheroids growth inhibition. The combination achieved the highest tumor growth suppression in vivo in the A549 xenograft. The combination of axitinib and doxorubicin exhibited the best anti-tumor effects both in vitro and in vivo than each single drug.

Phase I Study of Axitinib in Combination with Cisplatin and Capecitabine in Patients with Previously Untreated Advanced Gastric Cancer / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: This phase I trial evaluated the question of whether the standard starting dose of axitinib could be administered in combination with therapeutic doses of cisplatin/capecitabine in patients with previously untreated advanced gastric cancer, and assessed overall safety, pharmacokinetics, and preliminary antitumor activity of this combination. MATERIALS AND METHODS: Patients in dose level (DL) 1 received axitinib 5 mg twice a day (days 1 to 21) with cisplatin 80 mg/m2 (day 1) and capecitabine 1,000 mg/m2 twice a day (days 1 to 14) in 21-day cycles. Maximum tolerated dose (MTD) was the highest dose at which 5 consecutive days of missed axitinib due to thrombocytopenia. DL1 was established as the MTD, since higher DL cohorts were not planned. Common grade 3/4 non-hematologic adverse events in 22 patients treated at DL1 included hypertension (36.4%) and decreased appetite and stomatitis (18.2% each). Cisplatin/capecitabine slightly increased axitinib exposure; axitinib decreased capecitabine and 5-fluorouracil exposure. Eight patients (36.4%) each had partial response or stable disease. Median response duration was 9.1 months; median progression-free survival was 3.8 months. CONCLUSION: In patients with advanced gastric cancer, standard doses of axitinib plus therapeutic doses of cisplatin and capecitabine could be administered in combination. Adverse events were manageable.