Inhibition of anlotinib-induced autophagy attenuates invasion and migration by regulating epithelial-mesenchymal transition and cytoskeletal rearrangement through ATG5 in human osteosarcoma cells

Abstract The cure rates for osteosarcoma have remained unchanged in the past three decades, especially for patients with pulmonary metastasis. Thus, a new and effective treatment for metastatic osteosarcoma is urgently needed. Anlotinib has been reported to have antitumor effects on advanced osteosarcoma. However, both the effect of anlotinib on autophagy in osteosarcoma and the mechanism of anlotinib-mediated autophagy in pulmonary metastasis are unclear. The effect of anlotinib treatment on the metastasis of osteosarcoma was investigated by transwell assays, wound healing assays, and animal experiments. Related proteins were detected by western blotting after anlotinib treatment, ATG5 silencing, or ATG5 overexpression. Immunofluorescence staining and transmission electron microscopy were used to detect alterations in autophagy and the cytoskeleton. Anlotinib inhibited the migration and invasion of osteosarcoma cells but promoted autophagy and increased ATG5 expression. Furthermore, the decreases in invasion and migration induced by anlotinib treatment were enhanced by ATG5 silencing. In addition, Y-27632 inhibited cytoskeletal rearrangement, which was rescued by ATG5 overexpression. ATG5 overexpression enhanced epithelial-mesenchymal transition (EMT). Mechanistically, anlotinib-induced autophagy promoted migration and invasion by activating EMT and cytoskeletal rearrangement through ATG5 both in vitro and in vivo. Our results demonstrated that anlotinib can induce protective autophagy in osteosarcoma cells and that inhibition of anlotinib-induced autophagy enhanced the inhibitory effects of anlotinib on osteosarcoma metastasis. Thus, the therapeutic effect of anlotinib treatment can be improved by combination treatment with autophagy inhibitors, which provides a new direction for the treatment of metastatic osteosarcoma.

Effects of anlotinib on the malignant phenotype of glioma cells by mediating NF-κB signaling pathway / 中国药房

OBJECTIVE To investigate the effects of anlotinib on the malignant phenotype of glioma cells by regulating the nuclear factor-κB （NF-κB） signaling pathway. METHODS Human glioma T98G cells were cultured in vitro， and 5-fluorouracil was used as positive control to investigate the effects of different concentrations of anlotinib （5， 10， 20 μmol/L） on the ability of proliferation， adhesion， migration and invasion， the expressions of epithelial-mesenchymal transition （EMT） related proteins [E-cadherin， N-cadherin， vimentin and fibronectin （FN）]. NF- κB signaling pathway inhibitor （BAY 11-7082） and activator （prostratin） were additionally used to verify the possible mechanism of the above effects of anlotinib. RESULTS Anlotinib with 5， 10， 20 μmol/L could significantly decrease the activity of cell proliferation （except for 5 μmol/L anlotinib group）， migration rate， and the number of adherent cells and invasive cells， could significantly up-regulate the expression of E-cadherin protein while down-regulate the expressions of N-cadherin， vimentin and FN protein （P＜0.05）； the effect of 20 μmol/L anlotinib was similar to that of positive control （P＞0.05）. Compared with 10 μmol/L anlotinib， pathway inhibitor could significantly decrease the ability of proliferation， adhesion， migration and invasion， and the expressions of N-cadherin， vimentin， FN and phosphorylated NF-κB p65 protein， while could significantly up-regulate the expression of E-cadherin protein （P＜0.05）； above indexes were reversed significantly by pathway activator （P＜0.05）. CONCLUSIONS Anlotinib may inhibit the proliferation， adhesion， migration and invasion of human glioma T98G cells， which may be associated with the inhibition of the NF-κB signaling pathway， thus inhibiting cell EMT-like processes.

Efficacy of Combination of Camrelizumab with Anlotinib as Third-line Therapy for Patients with Advanced Non-small Cell Lung Cancer / 肿瘤防治研究

Objective To investigate the clinical efficacy and related adverse reactions of the combination of camrelizumab with anlotinib as the third-line therapy on advanced non-small cell lung cancer. Methods We retrospectively analyzed the clinical data of 84 patients with advanced non-small cell lung cancer after second-line treatment. According to different treatment methods, 44 patients who received camrelizumab combined with anlotinib were included in the observation group, and 40 patients who received anlotinib alone were included in the control group. The PFS, ORR, DCR and incidence of adverse reactions were analyzed and compared between the two groups. Results The median PFS of the observation group was longer than that of the control group (7.0 vs. 5.6 months, P=0.001). No statistically significant difference was observed in ORR, DCR, the incidence of adverse reactions or the incidence of adverse reactions above grade 3 between two groups (all P > 0.05). Conclusion The clinical efficacy of camrelizumab combined with anlotinib as third-line therapy on advanced non-small cell lung cancer is better than anlotinib alone, and the safety is good.

Prediction Model of Treatment Effect of Anlotinib on Extensive-stage Small Cell Lung Cancer Based on Combination of Disease and Syndrome Information / 肿瘤防治研究

Objective To construct a nomogram prediction model for the treatment effect of anlotinib with the participation of traditional Chinese medicine syndrome elements on the patients with extensive-stage small cell lung cancer (ES-SCLC) who previously received multiple lines of chemotherapy. Methods The clinical data of 127 patients with ES-SCLC who received at least two cycles of anlotinib treatment were retrospectively studied. Kaplan-Meier method was used to analyze the relationship between each factor and the overall survival time. Cox regression analysis was applied to screen the independent influencing factors of the prognosis of patients with ES-SCLC. R language was employed to build a nomogram prediction model, C-index was used to evaluate the model, and calibration curve was adopted to verify the accuracy of the model. Results Age, PS score, brain metastases, qi deficiency syndrome, yin deficiency syndrome, and blood stasis syndrome were related risk factors for ES-SCLC treated with anlotinib. PS score, brain metastasis, and blood stasis syndrome were independent prognostic factors. On the basis of these three independent influencing factors, a nomogram model was established to predict the prognosis of patients with ES-SCLC treated with anlotinib. The predicted risk was close to the actual risk, showing a high degree of coincidence. Conclusion The nomogram model established with PS score, blood stasis syndrome elements, and brain metastasis as independent factors can predict the prognosis of patients with ES-SCLC receiving second- and third-line treatment of anlotinib.

Effects of Decitabine Combined with Anlotinib on Proliferation and Apoptosis of Multiple Myeloma Cells / 中国实验血液学杂志

OBJECTIVE@#To investigate the biological effects and its relative mechanism of decitabine combined with anlotinib on multiple myeloma cells.@*METHODS@#The human MM cell lines and primary cells were treated with different concentrations of decitabine, anlotinib, and decitabine+anlotinib, respectively. The cell viability was detected and combination effect was calculated by CCK-8 assay. The apoptosis rate was measured by flow cytometry and the level of c-Myc protein was determined by Western blot.@*RESULTS@#Both decitabine and anlotinib could effectively inhibit the proliferation and induce the apoptosis of MM cell lines NCI-H929 and RPMI-8226. The effect of combined treatment on the inhibition of cell proliferation and induction of apoptosis was stronger than that of single-drug treatment. The combination of the two drugs also showed strong cytotoxicity in primary MM cells. Decitabine and anlotinib could down-regulate the level of c-Myc protein in MM cells and the c-Myc level in the combination group was the lowest.@*CONCLUSION@#Decitabine combined with anlotinib can effectively inhibit the proliferation and induce apoptosis of MM cells, which provides a certain experimental basis for the treatment of human MM.

PD-1 inhibitor plus anlotinib for metastatic castration-resistant prostate cancer: a real-world study / 亚洲男科学杂志(英文版)

Management and treatment of terminal metastatic castration-resistant prostate cancer (mCRPC) remains heavily debated. We sought to investigate the efficacy of programmed cell death 1 (PD-1) inhibitor plus anlotinib as a potential solution for terminal mCRPC and further evaluate the association of genomic characteristics with efficacy outcomes. We conducted a retrospective real-world study of 25 mCRPC patients who received PD-1 inhibitor plus anlotinib after the progression to standard treatments. The clinical information was extracted from the electronic medical records and 22 patients had targeted circulating tumor DNA (ctDNA) next-generation sequencing. Statistical analysis showed that 6 (24.0%) patients experienced prostate-specific antigen (PSA) response and 11 (44.0%) patients experienced PSA reduction. The relationship between ctDNA findings and outcomes was also analyzed. DNA-damage repair (DDR) pathways and homologous recombination repair (HRR) pathway defects indicated a comparatively longer PSA-progression-free survival (PSA-PFS; 2.5 months vs 1.2 months, P = 0.027; 3.3 months vs 1.2 months, P = 0.017; respectively). This study introduces the PD-1 inhibitor plus anlotinib as a late-line therapeutic strategy for terminal mCRPC. PD-1 inhibitor plus anlotinib may be a new treatment choice for terminal mCRPC patients with DDR or HRR pathway defects and requires further investigation.

Combination of anlotinib and irinotecan in the second-line treatment of metastatic colorectal cancer: a multicenter phase 1/2 trial / 中华肿瘤杂志

Objective: To evaluate the safety and efficacy of anlotinib plus irinotecan in the second-line treatment of patients with metastatic colorectal cancer (mCRC). Methods: This prospective phase 1/2 study was conducted in 2 centers in China (Cancer Hospital of Chinese Academy of Medical Sciences and Jiangsu Province Hospital). We enrolled patients with mCRC whose disease had progressed after first-line systemic therapy and had not previously treated with irinotecan to receive anlotinib plus irinotecan. In the phase 1 of the trial, patients received anlotinib (8 mg, 10 mg or 12 mg, po, 2 weeks on/1 week off) in combination with fixed-dose irinotecan (180 mg/m(2), iv, q2w) to define the maximum tolerated dose (MTD) and recommended phase 2 dose (RP2D). In the phase 2, patients were treated with the RP2D of anlotinib and irinotecan. The primary endpoints were MTD and objective response rate (ORR). Results: From May 2018 to January 2020, a total of 31 patients with mCRC were enrolled. Anlotinib was well tolerated in combination with irinotecan with no MTD identified in the phase 1, and the RP2D was 12 mg. Thirty patients were evaluable for efficacy analysis. Eight patients achieved partial response, and 21 had stable disease, 1 had progressive disease. The ORR was 25.8% and the disease control rate was 93.5%. With a median follow-up duration of 29.5 months, the median progression-free survival and overall survival were 6.9 months (95% CI: 3.7, 9.3) and 17.6 months (95% CI: 12.4, not evaluated), respectively. The most common grade 3 treatment-related adverse events (≥10%) were neutropenia (25.8%) and diarrhea (16.1%). There was no treatment-related death. Conclusion: The combination of anlotinib and irinotecan has promising anti-tumor activity in the second-line treatment of mCRC with a manageable safety profile.

Clinical observation of bevacizumab versus anlotinib respectively combined with chemotherapy drug in the treatment of EGFR-TKI acquired resistant advanced lung adenocarcinoma / 中国药房

OBJECTIVE To compare the short-term therapeutic effect and safety of bevacizumab versus anlotinib respectively combined with chemotherapy drug in the treatment of epidermal growth factor receptor tyrosine kinase inhibitors （EGFR-TKI） acquired resistant advanced lung adenocarcinoma. METHODS The information of 84 patients with EGFR-TKI acquired resistant advanced lung adenocarcinoma in the Third People’s Hospital of Chengdu was analyzed retrospectively during Jun. 2019-Oct. 2021. The patients were divided into chemotherapy group （32 cases）， anlotinib combined chemotherapy group （24 cases） and bevacizumab combined chemotherapy group （28 cases）. Patients in the chemotherapy group were given Pemetrexed disodium for injection and Carboplatin injection， and symptomatic treatment was given for adverse reactions. On the first day of chemotherapy， patients in the anlotinib combined chemotherapy group received Anlotinib hydrochloride capsules 10 mg orally， once a day， for 14 consecutive days and 7 days of discontinuation， based on the treatment of the chemotherapy group. Patients in the bevacizumab combined chemotherapy group were given Bevacizumab injection of 15 mg/kg intravenously 1 day before chemotherapy， based on the treatment of the chemotherapy group. Three groups of patients were treated for a total of four cycles， with one cycle every three weeks. The overall response rate （ORR）， disease control rate （DCR）， median progression-free survival （mPFS）， and the changes of serum tumor markers were compared among three groups before and after treatment； meanwhile， the occurrence of adverse drug reactions was recorded， and the 1-year survival rate was followed up. RESULTS After 4 treatment cycles， ORR and DCR of bevacizumab combined chemotherapy group and anlotinib combined chemotherapy group were higher than chemotherapy group （P＜0.05）； mPFS of the two groups were significantly longer than chemotherapy group， and DCR of anlotinib combined chemotherapy group was significantly higher than bevacizumab combined chemotherapy group （P＜0.05）. After 4 treatment cycles， the serum levels of tumor markers in three groups were significantly lower than before treatment， and both combined chemotherapy groups were significantly lower than chemotherapy group （P＜0.05）. There was no statistically significant difference in the incidence of adverse reactions such as nausea， vomiting， bone marrow suppression， and 1-year survival rate among the three groups of patients （P＞0.05）. CONCLUSIONS Bevacizumab and anlotinib combined with chemotherapy drug are effective and safe in the treatment of advanced lung adenocarcinoma with acquired EGFR-TKI resistance.

First line anlotinib plus liposomal doxorubicin for locally advanced or metastatic soft tissue sarcoma: A prospective, single-arm trial

Objective: To examine the efficacy and safety of anlotinib as firstline therapy to treat locally advanced or metastatic soft-tissue sarcoma. Methods: This is a single-arm trial. Treatment-naive patients (≥14 years) with locally advanced or metastatic soft tissue sarcoma were eligible. Each treatment cycle lasted for 3 weeks, and included liposomal doxorubicin (40-50 mg/m2) on day 1 and anlotinib (12 mg) on days 8-21. Starting from the 9th cycle, treatment consisted of only anlotinib. Treatment continued until disease progression or intolerable toxicities. The primary efficacy end point was progression-free survival (PFS). Results: Eight patients were enrolled between July 25, 2019 and January 8, 2020. The median number of treatment cycles was 5.5. Within 5.9 months median follow-up, PFS events occurred in 4 (4/8, 50%) patients. The median PFS was 11.3 months and the 6-month PFS rate was 56%. No patients attained complete response and 2 patients (fibrosarcoma, 1 patient and undifferentiated pleomorphic sarcoma, 1 patient) achieved partial response. Three patients (fibrosarcoma, 2 patients and synovial sarcoma, 1 patient) had stable disease. The objective response rate was 25% (2/8) for the study population, and the disease control rate was 75% (6/8). No new safety concerns emerged. Conclusions: Anlotinib plus liposomal doxorubicin demonstrated antitumor activities in previously untreated locally advanced or metastatic soft tissue sarcomas. Due to the small sample size, further investigations with a larger population should be undertaken to confirm the study findings.

Efficacy and Safety of Anlotinib Combined with PD-1 Blockades for Patients with Advanced Epithelial Ovarian Cancer / 肿瘤防治研究

Objective To investigate the efficacy and safety of anlotinib combined with PD-1 blockades for patients with advanced epithelial ovarian cancer (EOC). Methods A retrospective study was performed, enrolling 33 patients with advanced EOC who failed standard systematic therapy. All patients were administered with anlotinib combined with PD-1 blockades. The efficacy and safety profile were determined during treatment. Results The objective response rate of the 33 patients was 36.4% (95%CI: 20.4%-54.9%) and the disease control rate of the patients was 81.8% (95%CI: 64.5%-90.0%). The median PFS and OS of the 33-patient cohort were 7.6 months (95%CI: 3.1-12.1) and 19.6 months (95%CI: 15.1-24.1), respectively. The most common treatment-related adverse reactions were fatigue (66.7%), nausea and vomiting (54.5%), hypertension (48.5%), and diarrhea (39.4%). Furthermore, multivariate Cox regression analysis indicated that ECOG performance status and FIGO stage were independent factors for predicting the PFS of the combination regimen for patients with advanced EOC. Conclusion Anlotinib combined with PD-1 blockades preliminarily exhibit satisfactory efficacy and tolerable safety profile for patients with advanced EOC.

Effects of Tspan8 Gene Knockout Combined with Anlotinib on Proliferation, Migration, Invasion and Apoptosis of Colon Cancer SW480 Cells / 肿瘤防治研究

Objective To investigate the effects of Tspan8 gene knockout combined with anlotinib on the proliferation, migration, invasion, and apoptosis of colon cancer SW480 cells. Methods The plasmid was constructed by CRISPR/Cas9 technique, and Tspan8 gene was knocked out in SW480 cells. The knockout effect was detected by Western blot. The IC50 of anlotinib was calculated by MTT assay. The experiment was divided into control group, anlotinib group, Tspan8 knockout group, and combined group. Cell proliferation, migration, invasion, and apoptosis were detected by cell proliferation assay, clonal formation assay, scratch assay, Transwell chamber assay, and flow cytometry. Western blot was used to detect the effect of anlotinib on Tspan8 expression in SW480 cells. Results SW480-KO-Ⅲ cells had the highest knockout efficiency in the Tspan8 knockout group. They were used in subsequent experiments. Different concentrations of anlotinib could inhibit the proliferation of SW480 cells at different times (P < 0.01) in a concentration-dependent and time-dependent manner (P < 0.01). According to IC50, 14 μmol/L was selected as the subsequent experimental concentration. Compared with those in the control group, the cell proliferation, migration, and invasion abilities in the anlotinib group, Tspan8 knockout group, and combined group were significantly decreased, and the cell apoptosis level was significantly increased (P < 0.05). The above changes in the combined group were more significant than those in the anlotinib group or Tspan8 knockout group (P < 0.05). Compared with that in the control group, the expression level of Tspan8 in SW480 cells in the anlotinib group was significantly decreased (P < 0.01). Conclusion Tspan8 gene knockout combined with anlotinib can synergistically inhibit the proliferation, migration and invasion of SW480 cells. This combination can also promote the apoptosis of these cells.

A Case Report of AFP-producing Primary Lung Squamous Cell Carcinoma with Significant Therapeutic Effect / 中国肺癌杂志

BACKGROUND@#Primary lung squamous carcinoma that produces alpha-fetoprotein (AFP) is rare and only four related cases have been reported so far. The specific reasons for elevated serum level of AFP and effective treatment regimens for AFP-producing lung squamous carcinoma are not clear. This paper reports the diagnosis and treatment of AFP-producing lung squamous carcinoma so as to provide some references for similar cases in clinical practice.@*METHODS@#The diagnosis and treatment of an AFP-producing lung squamous carcinoma patient admitted to the Shandong Cancer Hospital on October 23, 2020 was retrospectively analyzed, and literatures were reviewed.@*RESULTS@#A 52-year-old male patient was diagnosed as T4N3M0 stage, IIIc right upper lobe lung squamous cell carcinoma with mediastinal lymph node metastasis and multiple metastases in the lung. The main tumor marker was abnormally increased serum AFP. After the rapid progression of two lines chemotherapy, the patient was given anlotinib combined with carrizumab as third-line treatment. The efficacy evaluation reached to partial response (PR) and stable disease (SD) after 2 and 4 cycles of treatment, respectively. The treatment regimen was replaced with albumin paclitaxel plus carrizumab due to gastrointestinal bleeding after the fifth cycle. The patient's condition was under continuous control.@*CONCLUSIONS@#The AFP-producing lung squamous carcinoma patient had a good response to anlotinib and immunotherapy in the case report, which may provide some guidances for the clinical practice and the research on AFP-producing lung squamous carcinoma.

Efficacy analysis of anlotinib combined with chemotherapy for advanced non-small cell lung cancer after failure of second-line chemotherapy / 国际肿瘤学杂志

Objective:To observe the efficacy and safety of anlotinib combined with chemotherapy in patients with advanced non-small cell lung cancer (NSCLC) who failed second-line chemotherapy.Methods:A retrospective analysis was performed on 80 patients with advanced NSCLC who had failed second-line chemotherapy admitted in the Department of Oncology of Chaohu Hospital of Anhui Medical University from January 2017 to October 2019, and the patients were divided into control group ( n=36) and observation group ( n=44) according to the different treatment regimens. The control group was given pemetrexed + cisplatin, and the observation group adopted pemetrexed + cisplatin + anlotinib. The objective response rate (ORR), disease control rate (DCR), median progression-free survival (PFS), overall survival (OS), changes in levels of serum vascular endothelial growth factor (VEGF), carcinoembryonic antigen (CEA) and carbohydrate antigen 199 (CA199) and treatment-related adverse reactions were compared between the two groups. Results:After 2 cycles of treatment, the ORR in the control group and observation group were 5.56% (2/36) and 18.18% (8/44), with no statistically significant difference ( χ2=1.85, P=0.174). The DCR in the two groups were 58.33% (21/36) and 81.82% (36/44), and the DCR in the observation group was significantly higher than that in the control group, with a statistically significant difference ( χ2=5.33, P=0.021). The median PFS in the two groups were 4.0 months and 6.0 months, and the median PFS in the observation group was longer than that in the control group, with a statistically significant difference ( χ2=28.47, P<0.001). The median OS in the two groups were 13.0 months and 14.8 months, with no statistically significant difference ( χ2=1.56, P=0.212). The levels of serum VEGF [(21.72±5.42) ng/L vs. (36.97±7.53) ng/L, t=14.13, P<0.001; (16.61±4.14) ng/L vs. (38.85±8.61) ng/L, t=23.09, P<0.001], CEA [(4.91±1.58) ng/ml vs. (6.62±2.84) ng/ml, t=4.64, P<0.001; (3.07±1.32) ng/ml vs. (7.08±3.31) ng/ml, t=11.50, P<0.001] and CA199 [(16.83±5.23) U/ml vs. (20.95±7.94) U/ml, t=3.75, P<0.001; (13.37±5.85) U/ml vs. (21.66±8.72) U/ml, t=7.55, P<0.001] in the control group and observation group after 2 cycles of treatment were significantly decreased compared with those before treatment, and the levels of serum VEGF, CEA and CA199 in the observation group were significantly lower than those in the control group ( t=4.78, P<0.001; t=5.68, P<0.001; t=2.76, P=0.007). The incidence of elevated blood pressure in the observation group was significantly higher than that in the control group [25.00% (11/44) vs. 2.78% (1/36), χ2=7.67, P=0.006]. Conclusion:Pemetrexed + cisplatin + anlotinib regimen for patients with advanced NSCLC who failed second-line chemotherapy can improve DCR, prolong PFS and improve the levels of serum tumor-related markers, with controllable adverse reactions.

Efficacy and influencing factors of anlotinib in treatment of elderly patients with small cell lung cancer after second-line treatment failure / 国际肿瘤学杂志

Objective:To investigate the efficacy and influencing factors of anlotinib in treatment of elderly patients with small cell lung cancer after second-line treatment failure.Methods:A total of 56 elderly patients with small cell lung cancer who were diagnosed and treated in the Tumor Hospital Affiliated to Xinjiang Medical University from September 2018 to February 2020 were collected. All patients were treated with anlotinib capsule after failure of second-line chemotherapy, objective response rate (ORR), disease control rate (DCR) and progression-free survival (PFS) were calculated, and ORR, DCR and PFS of patients with different clinical characteristics were compared. Cox proportional hazards model was used to analyze the factors influencing PFS in elderly patients with small cell lung cancer, and adverse drug reactions were observed.Results:After 2 cycles of treatment, the ORR and DCR of 56 elderly patients with small cell lung cancer were 10.7% (6/56) and 53.6% (30/56) respectively. Among them, the ORR and DCR of patients without brain metastasis were 20.8% (5/24) and 75.0% (18/24), which were higher than 3.1% (1/32) and 37.5% (12/32) in patients with brain metastasis, with statistically significant differences ( χ2=4.496, P=0.034; χ2=7.754, P=0.005). The ORR and DCR of patients with Eastern Cooperative Oncology Group (ECOG) score of 0-1 were 21.7% (5/23) and 69.6% (16/23), which were higher than those of patients with ECOG score of 2-3 [3.0% (1/33), 42.4% (14/33)], with statistically significant differences ( χ2=4.959, P=0.026; χ2=4.014, P=0.045). ORR and DCR were not related to gender, age, clinical stage or smoking status (all P>0.05). The median PFS of 56 patients was 3.8 months. The median PFS of patients aged ≤70 years was 5.0 months, and that of patients aged >70 years was 3.4 months, with a statistically significant difference ( χ2=5.452, P=0.020). The median PFS of patients without brain metastasis was 5.1 months, and that of patients with brain metastasis was 3.2 months, with a statistically significant difference ( χ2=8.895, P=0.003). The median PFS of patients with ECOG score of 0-1 was 5.0 months, and that of patients with ECOG score of 2-3 was 2.9 months, with a statistically significant difference ( χ2=5.923, P=0.015). The median PFS of patients with limited stage was 5.0 months, and that of patients with extensive stage was 3.1 months, with a statistically significant difference ( χ2=5.141, P=0.023). Cox multivariate analysis showed that ECOG score ( HR=2.522, 95% CI: 1.378-4.615, P=0.003) and brain metastasis or not ( HR=0.323, 95% CI: 0.168-0.622, P=0.001) were independent prognostic factors of PFS. During the treatment of anlotinib, the main adverse reactions were grade Ⅰ-Ⅱ, grade Ⅲ-Ⅳ adverse reactions were mainly hypertension and hand-foot syndrome, which improved after drug reduction and symptomatic treatment, and could be tolerated later. The incidence of drug reduction was 3.6% (2/56), and there were no patients with drug interruption or termination of treatment. Conclusion:Anlotinib has good short-term efficacy and survival benefits in the treatment of elderly patients with small cell lung cancer after second-line treatment failure. It has good therapeutic effect for patients with low ECOG score and without brain metastasis, and has tolerable adverse reactions and high safety.

Efficacy and Safety of PD-1 Inhibitor Combined with Anlotinib on Advanced Neuroendocrine Carcinoma / 肿瘤防治研究

Objective To analyze the efficacy and safety of PD-1 inhibitor combined with anlotinib on advanced neuroendocrine carcinoma. Methods We collected the data of patients with advanced neuroendocrine carcinoma who had failed the first-line standard chemotherapy and treated with PD-1 inhibitor combined with anlotinib from the First Affiliated Hospital of Zhengzhou University. Results A total of 45 patients, including 24 males and 21 females, were included. The median age was 57 years old. The primary tumor sites were lung (23 cases, 51.1%), esophagus (8 cases, 17.8%), pancreas (7 cases, 15.6%) and rectum (7 cases, 15.6%). Eighteen cases (40%) had failed the first- and second-line treatments, and 27 cases (60%) had failed the third-line and above treatments. All patients received 2-15 cycles of treatment, 3 cases died due to disease progression, overall objective response rate was 11.1%, disease control rate was 53.5%, median progression-free survival was 5.8 months, and 10-month progression-free survival rate was 25.5%. Adverse events were mainly grade 1-2 myelosuppression and digestive tract reactions. Conclusion PD-1 combined with anlotinib show better efficacy and good tolerance on advanced neuroendocrine carcinoma. It can be used as a choice after the failure of standard first-line treatment of advanced neuroendocrine carcinoma.

Clinical Efficacy of Anlotinib Combined with Paclitaxel and Cisplatin as First-line Treatment of Advanced Esophageal Squamous Cell Carcinoma / 肿瘤防治研究

Objective To observe the efficacy and side effects of anlotinib combined with paclitaxel and cisplatin as the first-line treatment of advanced esophageal squamous cell carcinoma (ESCC). Methods We retrospectively analyzed 50 cases of advanced esophageal squamous cell carcinoma diagnosed pathologically. Among them, 24 cases were treated with the combination of anlotinib and paclitaxel plus cisplatin (experimental group), and 26 cases were treated with paclitaxel plus cisplatin regimen (control group). The efficacy and adverse reactions were observed and followed up. Results The objective response rates of the experimental and control groups were 83.33% and 53.84% (P < 0.05), the disease control rates were 100% and 96.15% (P > 0.05), mPFS were 10.6 and 9.13 months (P < 0.05), and mOS were 13.4 and 11.8 months (P < 0.05). The common adverse events in the experimental group were hand-foot syndrome (12.5%), hypertension (12.5%), epistaxis (8.33%) and proteinuria (4.16%), all of which were grade Ⅰ-Ⅱ and controllable without affecting the continuity of chemotherapy. Conclusion The combination of anlotinib, paclitaxel and cisplatin as the first-line treatment of advanced esophageal squamous cell carcinoma can improve the curative effect and the adverse effects are endurable.

Clinical study of anlotinib combined with irinotecan in the third line treatment of metastatic esophageal cancer / 国际肿瘤学杂志

Objective:To investigate the clinical efficacy and adverse reactions of anlotinib combined with irinotecan in the third line treatment of metastatic esophageal cancer.Methods:From October 2018 to October 2019, 52 patients with metastatic esophageal cancer who had developed distant metastasis after receiving standard concurrent chemoradiotherapy and failed second-line chemotherapy were selected from Lu′an Hospital of Traditional Chinese Medicine of Anhui Province. The patients were divided into experimental group and control group by random number table method, with 26 cases in each group. The control group was given intravenous chemotherapy with irinotecan. The experimental group was treated with oral erlotinib combined with intravenous chemotherapy of irinotecan. The clinical efficacy and adverse reactions of the two groups were evaluated after 2 cycles of treatment.Results:Before treatment, there was no significant difference in Karnofsky performance status (KPS) score between the experimental group and the control group (76.15±7.52 vs. 74.62±8.59, t=-0.137, P=0.892). After treatment, there was no significant difference between the two groups (70.77±6.28 vs. 72.69±8.74, t=-1.761, P=0.084). However, after treatment, the KPS score in the experimental group was lower than that before treatment ( t=3.035, P=0.006). There was no statistical significance in the KPS scores of the control group before and after treatment ( t=1.000, P=0.327). Adverse reactions in the two groups were mainly grade 1-2. The incidences of grade 1-2 myelosuppression and diarrhea in the experimental group were 61.5% (16/26) and 46.2% (12/26), which were significantly higher than those in the control group (19.2%, 5/26 and 19.2%, 5/26), with statistically significant differences ( χ2=9.665, P=0.002; χ2=4.282, P=0.039). The disease control rate of the experimental group was 73.1% (19/26), which was significantly higher than that of the control group (46.2%, 12/26), and there was a statistically significant difference between the two groups ( χ2=3.914, P=0.048). The median progression-free survival of the experimental group and the control group was 52 days and 45 days, respectively, and there was a statistically significant difference ( χ2=4.692, P=0.032). Conclusion:Anlotinib combined with irinotecan in the third-line treatment of metastatic esophageal cancer has obvious efficacy, but to a certain extent, it increases the incidence of grade 1-2 myelosuppression and diarrhea, and the KPS score is lower compared with before treatment.

Clinical observation of 125I seed implantation combined with amlotinib hydrochloride in the treatment of advanced non-small cell lung cancer / 国际生物医学工程杂志

Objective:To investigate the clinical efficacy of 125I seed implantation combined with anlotinib hydrochloride in the treatment of non-small cell lung cancer (NSCLC). Methods:61 cases of NSCLC patients were enrolled, of which 30 cases (observation group) received 125I seed implantation combined with anlotinib treatment, and 31 cases (control group) received 125I seed implantation only. To evaluate the curative effect and adverse reactions of all patients, the carcinoembryonic antigen (CEA), cytokeratin 19 fragment (CYFRA21-1), neuroendocrine enolase (NSE), squamous cell carcinoma antigen (SCC) in the peripheral blood of the two groups was measured before the treatment and at 1 and 3 months after the treatment. Results:The effective rates in the observation group were 90.00% and 93.33%, the effective rates in the control group were 67.74% and 74.19% at 1 and 3 months after the treatment, respectively, and the difference in efficacy between the two groups was statistically significant ( χ2=4.504, P=0.034 vs. χ2=4.075, P=0.044). There was no significant difference in the incidence of adverse reactions between the two groups of patients after treatment ( P=0.785). At 1 and 3 months after the treatment, the levels of CEA, CYFRA21-1, NSE and SCC in the peripheral blood of the two groups of patients were lower than those before the treatment (all P<0.05). Conclusions:125I seed implantation combined with anlotinib hydrochloride is safe for the treatment of advanced non-small cell lung cancer, and has promotion value.

Observation of short-term efficacy of bronchial arterial chemoembolization combined with anlotinib for treatment of advanced non-small cell lung cancer / 肿瘤研究与临床

Objective:To observe the short-term efficacy and safety of bronchial arterial chemoembolization (BACE) combined with anlotinib for treatment of advanced non-small cell lung cancer (NSCLC).Methods:The clinical data of 14 patients with advanced NSCLC in the First Affiliated Hospital of Zhengzhou University from June 2018 to March 2019 were retrospectively analyzed. The short-term efficacy and adverse reactions of BACE combined with anlotinib hydrochloride were evaluated.Results:All patients successfully received BACE treatment twice. The median follow-up time was 19 months (8-26 months). The objective response rate (ORR) of patients at 1, 3 and 6 months after the first treatment was 100.0% (14/14), 71.4% (10/14) and 57.1% (8/14), and the disease control rate (DCR) was 100.0% (14/14), 92.8% (13/14) and 78.6% (11/14), respectively. The median progression-free survival (PFS) time was 9.5 months (95% CI 9.0-17.3 months), and the 6-month and 12-month PFS rates were 78.6% and 28.6%, respectively. The median overall survival (OS) time was 19.0 months (95% CI 18.4-23.1 months), and the 6-month and 12-month OS rates were 100.0% and 85.7%, respectively. Anlotinib hydrochloride-related adverse reactions included hand-foot syndrome [42.9% (6/14)], fatigue [35.7% (5/14)], hypertension [35.7% (5/14)], oral mucositis [28.6% (4/14)], hemoptysis [28.6% (4/14)], elevated aminotransferases [21.4% (3/14)] and diarrhea [14.3% (2/14)]. There were no grade ≥3 adverse reactions. Conclusion:BACE combined with anlotinib is safe and effective for treatment of advanced NSCLC, and the short-term clinical efficacy is satisfactory.

Establishment and Application of Method for Concentration Determination of Anlotinib in Human Plasma / 中国药房

OBJECTIVE：To establish a method for concentration determination of anlotinib in human plasma and apply it in the clinic. METHODS ：The plasma samples were pretreated by salting-out assisted with liquid-liquid extraction with ammonium acetate as salting out assistant and acetonitrile as solvent. Using voriconazole as internal standard ，LC-MS/MS method was adopted. The separation was performed on Waters X Bridge C 18 column with mobile phase consisting of 0.2％ formic acid solution- acetonitrile（gradient elution ）at the flow rate of 1 mL/min. The column temperature was set at 40 ℃，and sample size was 10 μL. The split ratio was 3∶7. The electrospray ion source and multiple reaction monitoring mode were used for the analysis. The ion pair of anlotinib and internal standard under positive ion mode were m/z 408.3→339.3 and m/z 350.2→281.3，respectively. RESULTS ： Anlotinib showed a good linear relationship in the concentration range of 0.2-200 ng/mL（R2＞0.996 7）. The lowest limit of quantitation was 0.2 ng/mL. Intra-day and inter-day RSDs were no more than 12％ （n＝6 or n＝3）. Accuracies were 90.92％-108.00％（n＝6 or n＝3）. The average extraction recoveries were 87.51％-100.00％（RSD＜8％，n＝6）. The average matrix effects were 96.66％-99.93％（RSD＜5％，n＝6）. The plasma concentration of 3 patients with NSCLC treated with anlotinib was 8.74-65.60 ng/mL. CONCLUSIONS ：The method is simple ，accurate and specific ，and is suitable for the plasma concentration monitoring of anlotinib in NSCLC patients.