Journal: Discover Oncology

Article Title: Anlotinib inhibits esophageal cancer malignancy by ameliorating the immune microenvironment

doi: 10.1007/s12672-026-04457-8

Figure Lengend Snippet: Anlotinib inhibited the proliferation of EC cells both in vitro andin vivo. A , B Anlotinib sensitivity assay showsthat the value of IC50 of ECA109 cells is 2992 nM and the value of IC50 of KYSE150 cells is 2279 nM. C , D Cell survivalassay shows that anlotinib can inhibit the growth of ECA109 cells and KYSE150 cells treated with 2.5μmol anlotinib. E Representative xenografted tumors from Anlotinib group and the control group. F The curves of tumor volumesof mice in the Anlotinib group and the control group as indicted time. G The tumor weight in the Anlotinib groupis significantly smaller than that in the control group. H The curves of body weights of mice in the Anlotinib groupand the control group as indicted time time. The t test was used for statistic quantifications: *p < 0.05, ***p < 0.001,****p < 0.0001

Article Snippet: Anlotinib (AL3818) dihydrochloride was purchased from Selleck, and its stock solution was prepared according to the manufacturer’s instructions.

Techniques: In Vitro, Sensitive Assay, Control

Journal: Discover Oncology

Article Title: Anlotinib inhibits esophageal cancer malignancy by ameliorating the immune microenvironment

doi: 10.1007/s12672-026-04457-8

Figure Lengend Snippet: Anlotinib regulated immune infiltration through VEGFR2. A The mRNA level of VEGFR2 was examined by quantitative PCR. B The expression of marker genes related to immune cells was examined by quantitative PCR. C Analysis of public databases shows that the high expression of VEGFR2 is significantly associated with a high overall survival rate in anti-PD-1, PD-L1 and CTLA-4 immunotherapies. D Analysis of public databases shows that the high expression of VEGFR2 is significantly associated with a high disease-free survival rate in anti-PD-1, PD-L1 and CTLA-4 immunotherapies. The t test was used for statistic quantifications: *** p < 0.001, ns, not significant.

Article Snippet: Anlotinib (AL3818) dihydrochloride was purchased from Selleck, and its stock solution was prepared according to the manufacturer’s instructions.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Marker

Journal: Pharmaceuticals

Article Title: Concept of Hybrid Drugs and Recent Advancements in Anticancer Hybrids

doi: 10.3390/ph15091071

Figure Lengend Snippet: Current status of indole-based hybrids that are approved or/-under clinical trials.

Article Snippet: Chia-tai Tianqing Pharmaceutical Co. , AL 3818 (Anlotinib) , Tyrosine kinase , Synovial sarcoma, Advanced alveolar soft part sarcoma , Clinical trials , [ ] .

Techniques: Clinical Proteomics, Histone Deacetylase Assay

Journal: Frontiers in Oncology

Article Title: Targeting liposarcoma: unveiling molecular pathways and therapeutic opportunities

doi: 10.3389/fonc.2024.1484027

Figure Lengend Snippet: Clinical trials related to LPS-targeted therapy.

Article Snippet: ALTER-0202 (NCT01878448) , Anlotinib , A multikinase angiogenesis inhibitor , II , 166 STS pts , LPS (n = 13) , PFR12 weeks , – , LPS PFR 12weeks:63% LPS mPFS:5.6m LPS mOS:13 m LPS ORR:7.7% , Yihebali Chi , 2018 , ( ) .

Techniques: Clinical Proteomics, Activity Assay

Journal: Frontiers in Oncology

Article Title: Targeting liposarcoma: unveiling molecular pathways and therapeutic opportunities

doi: 10.3389/fonc.2024.1484027

Figure Lengend Snippet: Clinical trials of LPS-targeted and immunotherapy are currently underway.

Article Snippet: ALTER-0202 (NCT01878448) , Anlotinib , A multikinase angiogenesis inhibitor , II , 166 STS pts , LPS (n = 13) , PFR12 weeks , – , LPS PFR 12weeks:63% LPS mPFS:5.6m LPS mOS:13 m LPS ORR:7.7% , Yihebali Chi , 2018 , ( ) .

Techniques: Clinical Proteomics, Bioprocessing, Blocking Assay, Expressing, Activity Assay

Journal: Thoracic Cancer

Article Title: Occurrence of hypertension during third‐line anlotinib is associated with progression‐free survival in patients with squamous cell lung cancer ( SCC ): A post hoc analysis of the ALTER0303 trial

doi: 10.1111/1759-7714.14076

Figure Lengend Snippet: Clinical characteristics of the SCC patients enrolled in the study

Article Snippet: All patients received oral anlotinib (12 mg/day on days 1–14 of a 21‐day cycle.) (Chia Tai Tianqing Pharmaceutical Group Co., Ltd) or placebo (Chia Tai Tianqing Pharmaceutical Group Co., Ltd) until progression, unacceptable toxicity, withdrawal of patient consent, or death.

Techniques:

Journal: Annals of Translational Medicine

Article Title: Anlotinib suppresses lung adenocarcinoma growth via inhibiting FASN-mediated lipid metabolism

doi: 10.21037/atm-22-5438

Figure Lengend Snippet: Anlotinib regulated lipid metabolism in A549 mice xenograft models. (A) The mass volumes of A549 subcutaneous xenografts in nude mice after treatment with placebo or anlotinib (0.8 mg/kg) (n=7 per group). (B) The picture of excited xenograft tumors from both control and anlotinib groups. (C) Body weights of mice in both groups are shown (n=7 per group). (D) Total counts of up-regulated/down-regulated proteins by anlotinib through proteomic profiling. (E) Metabolism-related pathways enrichment analyzed by KEGG analysis. (F) The visualization of the relative fold changes of lipid metabolism-related proteins between anlotinib group and placebo group by heatmap. (G) GC-MS analysis of medium and long chain fatty acids in A549 cells with indicated treatment for 72 h. (H) The neutral lipid droplet staining with Bodipy 493/503 in A549 cells with indicated treatment for 48 h (×120). Red arrowheads indicate neutral lipid droplets. Data are expressed as mean ± SEM. ****, P<0.0001; ns, not significant. KEGG, Kyoto Encyclopedia of Genes and Genomes; GC-MS, gas chromatography-mass spectrometry; SEM, standard error of the mean.

Article Snippet: Anlotinib powder was kindly provided by Chiatai Tianqing (CTTQ) pharma (Jiangsu, China).

Techniques: Control, Gas Chromatography-Mass Spectrometry, Staining, Gas Chromatography, Mass Spectrometry

Journal: Annals of Translational Medicine

Article Title: Anlotinib suppresses lung adenocarcinoma growth via inhibiting FASN-mediated lipid metabolism

doi: 10.21037/atm-22-5438

Figure Lengend Snippet: Anlotinib suppressed lung adenocarcinoma cells through FASN. (A) RT-qPCR analysis of lipid metabolism-related genes in A549 cells after indicated treatments for 72 h. (B) Upper panel: Western blotting analysis of FASN expression as anlotinib concentration increased. Lower Panel: the protein quantification results analyzed by ImageJ software. (C) FASN expression levels of A549-shFASN, A549-shNC, A549-oeNC, and A549-oeFASN cells as determined by western blot. (D) Upper panel: colony formation assay of A549-shFASN, A549-shNC, A549-oeNC, and A549-oeFASN cells treated with anlotinib at the indicated concentrations for 10 days. Lower panel: the colony formation rate, computed by calculating the ratio of clone numbers between indicated group and the untreated negative control (stained with 0.5% crystal violet for 30 min). (E) CCK-8 assay of A549-shFASN, A549-shNC, A549-oeNC, and A549-oeFASN cells treated with anlotinib at the indicated concentrations for 48 h. Data were expressed as mean ± SEM. *, P<0.05; **, P<0.01; ****, P<0.0001; ns, not significant. RT-qPCR, reverse transcription-quantitative polymerase chain reaction; FASN, fatty acid synthase; CCK-8, cell counting kit-8; SEM, standard error of the mean.

Article Snippet: Anlotinib powder was kindly provided by Chiatai Tianqing (CTTQ) pharma (Jiangsu, China).

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Concentration Assay, Software, Colony Assay, Negative Control, Staining, CCK-8 Assay, Reverse Transcription, Real-time Polymerase Chain Reaction, Cell Counting

Journal: Annals of Translational Medicine

Article Title: Anlotinib suppresses lung adenocarcinoma growth via inhibiting FASN-mediated lipid metabolism

doi: 10.21037/atm-22-5438

Figure Lengend Snippet: Anlotinib exhibited promising antitumor effects in an advanced lung adenocarcinoma patient with multiple treatment failure. (A) The timeline for entire treatment. Staging was performed as per AJCC 7th edition guideline. (B,C) CT images at baseline ahead of anlotinib treatment. (D,E) CT images at best response. Arrowheads with a same color indicate a same target lesion. AJCC, American Joint Committee on Cancer; CT, computed tomography; ARMS-PCR, amplification refractory mutation system polymerase chain reaction; EGFR, epidermal growth factor receptor; PD, progressive disease; NGS, next-generation sequencing.

Article Snippet: Anlotinib powder was kindly provided by Chiatai Tianqing (CTTQ) pharma (Jiangsu, China).

Techniques: Computed Tomography, Amplification, Mutagenesis, Polymerase Chain Reaction, Next-Generation Sequencing

Journal: Annals of Translational Medicine

Article Title: Anlotinib suppresses lung adenocarcinoma growth via inhibiting FASN-mediated lipid metabolism

doi: 10.21037/atm-22-5438

Figure Lengend Snippet: Anlotinib elicited robust antitumor effects and decreased FASN expression in a PDX model. (A) Schematic model presenting the procedure to construct the PDX model. (B) The tumor volume changes of PDX model iterations F1-F4 post inoculation. (C) H&E and IHC pictures of tumors from the established F4 PDX model and the parental tissue from the patient (×200). (D,E) The tumor volumes (D) and body weights (E) of PDX model mice treated with control or anlotinib (n=5 per group). (F) IHC staining of FASN in PDX models with or without treatment of anlotinib. Left panel shows the representative pictures, and panel is the calculated IRS. Data are expressed as mean ± SEM. *, P<0.05; ****, P<0.0001; ns, not significant as compared with the control group. NOD/SCID, anesthetized nonobese diabetic/severe combined immunodeficient; FASN, fatty acid synthase; PDX, patient-derived xenograft; IHC, immunohistochemical; IRS, immunoreactive score; H&E, hematoxylin and eosin staining; SEM, standard error of the mean.

Article Snippet: Anlotinib powder was kindly provided by Chiatai Tianqing (CTTQ) pharma (Jiangsu, China).

Techniques: Expressing, Construct, Control, Immunohistochemistry, Derivative Assay, Immunohistochemical staining, Staining

Journal: Cell Death & Disease

Article Title: Directly targeting c-Myc contributes to the anti-multiple myeloma effect of anlotinib

doi: 10.1038/s41419-021-03685-w

Figure Lengend Snippet: A Chemical structure of anlotinib. B MM cell lines (NCI-H929, RPMI-8226, LP1, MM.1S, OPM2, and U266) were treated with anlotinib (0–20 μM) for 48 h, and the cell viability was detected by CCK8. The IC50 value was calculated based on the dose–response curve using Prism 8.0 software. C – E NCI-H929, RPMI-8226 and LP1 cells were treated with anlotinib for 24 and 48 h, followed by the assessment of cell viability. F The CD138 + and G CD138 − cells isolated from MM patients were treated with anlotinib (0–10 μM) for 24 h, and then the cell viability was assessed. H Normal BMMCs from four healthy donors were treated with anlotinib (0–10 μM) for 24 h. Data are shown as mean ± SD and representative of three independent experiments.

Article Snippet: Anlotinib, CHX and MG-132 were purchased from CSNpharm (Chicago, IL, USA), dissolved in Dimethyl Sulfoxide (Sigma, St. Louis, MO) and stored in dark at −20 °C until use.

Techniques: Software, Isolation

Journal: Cell Death & Disease

Article Title: Directly targeting c-Myc contributes to the anti-multiple myeloma effect of anlotinib

doi: 10.1038/s41419-021-03685-w

Figure Lengend Snippet: A , B NCI-H929 and RPMI-8226 cells were co-cultured with or without BMSCs, and then treated with anlotinib (0–10 μM) for 48 h. After staining with Annexin V and PI, flow cytometry analysis was performed to assess the apoptosis rate. C , D NCI-H929 and RPMI-8226 cells were treated with anlotinib (0–10 μM) for 48 h, in the presence or absence of IL-6 (10 ng/ml). Cell proliferation was measured by CCK-8 assay. ** P < 0.01; *** P < 0.001; NS P > 0.05. Data are shown as mean ±SD and from three independent experiments.

Article Snippet: Anlotinib, CHX and MG-132 were purchased from CSNpharm (Chicago, IL, USA), dissolved in Dimethyl Sulfoxide (Sigma, St. Louis, MO) and stored in dark at −20 °C until use.

Techniques: Cell Culture, Staining, Flow Cytometry, CCK-8 Assay

Journal: Cell Death & Disease

Article Title: Directly targeting c-Myc contributes to the anti-multiple myeloma effect of anlotinib

doi: 10.1038/s41419-021-03685-w

Figure Lengend Snippet: A NCI-H929 and RPMI-8226 cells were treated with 5 µM anlotinib for the indicated time. Cell cycle was analyzed by flow cytometry. The numerical percentage shown indicates the G 2 /M population. B The morphology of NCI-H929 and RPMI-8226 cells after anlotinib treatment (5 μM, 24 h) was observed by Wright staining. Scale bar: 20 μm. C Representative images of TUNEL and DAPI staining for cells treated with anlotinib (5 μM, 24 h). The green dot represents positive TUNEL staining. The blowup images from the red boxes showed a significant fraction of nuclei after anlotinib exposure. Scale bar: 20 μm. D NCI-H929 and RPMI-8226 cells were treated with anlotinib (0–10 μM) for 24 h. Whole cell lysates were subjected to western blotting using antibodies against PARP-1, caspase 3, caspase 9, and β-actin. E Cells were treated as described in D , and apoptosis was detected by flow cytometry using Annexin V/PI staining. The percentages of apoptotic cells were shown in the histogram from three independent experiments. Each experiment was performed in triplicate. Con: control group; Anlo: anlotinib group. *** P < 0.001.

Article Snippet: Anlotinib, CHX and MG-132 were purchased from CSNpharm (Chicago, IL, USA), dissolved in Dimethyl Sulfoxide (Sigma, St. Louis, MO) and stored in dark at −20 °C until use.

Techniques: Flow Cytometry, Wright Stain, TUNEL Assay, Staining, Western Blot, Control

Journal: Cell Death & Disease

Article Title: Directly targeting c-Myc contributes to the anti-multiple myeloma effect of anlotinib

doi: 10.1038/s41419-021-03685-w

Figure Lengend Snippet: A Volcano plot of the DEGs from the transcriptomes of the control and anlotinib group ( P < 0.05, Fold change > 1.5). B KEGG analysis of the DEGs. C NCI-H929 and RPMI-8226 cells were treated with anlotinib (5 μM) for the indicated time. Whole cell lysates were subjected to western blotting. D Heatmaps of the top 40 downregulated c-Myc target genes in NCI-H929 cells treated with anlotinib versus DMSO for 12 h. Rows show Z -scores are calculated for each cell type. E NCI-H929, RPMI-8226, and LP1 cells were treated with the indicated concentration of anlotinib for 24 h or 5 μM anlotinib for the indicated time. Whole cell lysates were subjected to western blotting using c-Myc and β-actin antibodies. Con control group, Anlo anlotinib group. The experiments were performed in triplicate.

Article Snippet: Anlotinib, CHX and MG-132 were purchased from CSNpharm (Chicago, IL, USA), dissolved in Dimethyl Sulfoxide (Sigma, St. Louis, MO) and stored in dark at −20 °C until use.

Techniques: Control, Western Blot, Concentration Assay

Journal: Cell Death & Disease

Article Title: Directly targeting c-Myc contributes to the anti-multiple myeloma effect of anlotinib

doi: 10.1038/s41419-021-03685-w

Figure Lengend Snippet: A NCI-H929 cells were treated with CHX (10 μg/ml) in the presence or absence of anlotinib (5 μM) for the indicated time. The levels of c-Myc protein were evaluated and plotted in the graph. B NCI-H929 and RPMI-8226 cells were treated with either anlotinib (5 μM) and/or MG132 (5 μM) for 6 h, and then the c-Myc protein was assessed by western blotting. C NCI-H929 cells were pretreated with 5 μM MG132 for 3 h and then incubated with 5 μM anlotinib for 3 h. The cell lysates were immunoprecipitated with anti-c-Myc antibody and then probed with anti-ubiquitin antibody. D , E The binding between anlotinib and c-Myc protein was examined by the CETSA method at different temperatures or doses. The indicated proteins were evaluated by western blotting (left). CETSA curves of c-Myc were determined in the absence and presence of anlotinib. Each band intensity of c-Myc in DMSO or anlotinib group was normalized with respect to that obtained at the lowest temperature or dose (right). F Whole cell lysates of NCI-H929 cells were incubated with anlotinib followed by digestion with pronase according to the “Materials and methods” section. Then, the degree of c-Myc degradation was determined by western blot. G , H Overexpression and shRNA knockdown efficiency of c-Myc in NCI-H929 cells was measured by western blotting analysis. The effects of c-Myc knockdown or overexpression on cell apoptosis were evaluated by flow cytometry. The presented columns are given as the means ± SD. Statistically significant values compared with the DMSO group or empty vector group are depicted by * or # , respectively. *** P < 0.001, # P < 0.05, and ### P < 0.001. Con control group, Anlo anlotinib group. Data are shown as mean ± SD and representative of three independent experiments.

Article Snippet: Anlotinib, CHX and MG-132 were purchased from CSNpharm (Chicago, IL, USA), dissolved in Dimethyl Sulfoxide (Sigma, St. Louis, MO) and stored in dark at −20 °C until use.

Techniques: Western Blot, Incubation, Immunoprecipitation, Ubiquitin Proteomics, Binding Assay, Over Expression, shRNA, Knockdown, Flow Cytometry, Plasmid Preparation, Control

Journal: Cell Death & Disease

Article Title: Directly targeting c-Myc contributes to the anti-multiple myeloma effect of anlotinib

doi: 10.1038/s41419-021-03685-w

Figure Lengend Snippet: A NCI-H929, NCI-H929-BR, MM.1S, and MM.1S-BR cells were treated with various concentrations of bortezomib for 24 h. Cell viability was measured by CCK8 assay. B NCI-H929-BR and MM.1S-BR cells were treated with anlotinib (0–20 µM) for 24 and 48 h, followed by an assessment of cell viability. C NCI-H929-BR and MM.1S-BR cells were treated with 5 µM anlotinib for 8 h. The cell cycle was analyzed by flow cytometry. D The morphology of NCI-H929-BR and MM.1S-BR cells after anlotinib treatment (5 μM, 8 h) was observed using Wright staining. E Apoptosis of cells treated with 5 µM anlotinib for 24 h was detected by flow cytometry using Annexin V/PI staining. F NCI-H929-BR and MM.1S-BR cells were treated with anlotinib (5 μM) for the indicated time. Whole cell lysates were subjected to western blotting. G The combined effects of anlotinib and bortezomib in NCI-H929 cells were assessed using the CompuSyn software. Con: control group; Anlo: anlotinib group. *** P < 0.001. Each experiment was performed in triplicate.

Article Snippet: Anlotinib, CHX and MG-132 were purchased from CSNpharm (Chicago, IL, USA), dissolved in Dimethyl Sulfoxide (Sigma, St. Louis, MO) and stored in dark at −20 °C until use.

Techniques: CCK-8 Assay, Flow Cytometry, Wright Stain, Staining, Western Blot, Software, Control

Journal: Cell Death & Disease

Article Title: Directly targeting c-Myc contributes to the anti-multiple myeloma effect of anlotinib

doi: 10.1038/s41419-021-03685-w

Figure Lengend Snippet: A Nude mice bearing subcutaneous NCI-H929 tumors were treated with either anlotinib (3 mg/kg) or vehicle control by intragastric administration for consecutive 14 days. Tumor size was measured every 2 days. B The tumor tissues were excised and weighed on day 14. C Gross appearance of the tumors. D The weight of mice was monitored every 2 days. E Representative IHC staining of tumor tissues. Scale bars: 50 μM. F Tumor tissues were lysed and subjected to western blotting to detect the protein level of c-Myc. Con control group, Anlo anlotinib group. ** P < 0.01, *** P < 0.001.

Article Snippet: Anlotinib, CHX and MG-132 were purchased from CSNpharm (Chicago, IL, USA), dissolved in Dimethyl Sulfoxide (Sigma, St. Louis, MO) and stored in dark at −20 °C until use.

Techniques: Control, Immunohistochemistry, Western Blot