Service Overviews Background：In vivo pharmacodynamic evaluation plays a vital role in basic research and therapeutic drug development. Pharmacodynamic research can help define the clinical potential of a new drug and identify the strengths and weaknesses in comparison to other drugs already on the market. Animal tumor models focus on spontaneous tumors transplanted models. Transplanted tumors have the advantages of definite characteristics: rapid and reliable tumor growth short experimental period, models for various types. Methods：Alternative animal models can be applied as cost-effective research tools to conduct intensive and reliable experiments for personalized research purposes. Advantages：Prefabricated models can be completed in 4 weeks from the bearing of tumor to the end of administration, and we have abundant antibodies and cell lines. We are committed to offer batch-to batch consistency in cell resources with high stability and low within-group variation. Cases: Since the establishment of Sanyou Bio in vivo platform in 2018, we have been committed to provide you with 150+ small animal models and 56 days of in vivo efficacy rapid efficiency evaluation services. By April 2022, we have successfully developed 60+ tumor models, covering major tumors, and we have successfully completed efficacy evaluation of a series of targets such as TNFR2 and CD39, and accumulated extensive project experience for different molecular types. Service Contents Services Service Details Client Provides Deliverables and Standards Time Efficacy evaluation (not prefabricated) 1. Cell line construction 2. Cell line validation 3. STR analysis (Optional) 4. Tumor growth curve 5. Model development 6. Efficiency evaluation 1. Target MOA 2. Antibody 3. Experimental scheme 1. Quality report 2. Tumor growth curve 3. Report 6-8 weeks Efficacy evaluation (prefabricated) Efficacy evaluation 1. Target MOA 2. Antibody Report ~4 weeks Multicolor flow Cytometry Tissue/blood cell subtype flow cytometry Biomarker Report 1~3 days Cytokine/enzyme detection 1. Multi/single cytokines 2.Enzymes, etc Biomarker Report 1~3 days Pharmacokinetic analysis 1.PK experiment 2.PK analysis 1. Target MOA 2. Antibody Report 45 days Service Highlights 1. A Wide Array of Mouse Strains and Disease Models Sanyou Bio has a wide array of transgenic mice and different immunodeficient mice to meet your demands, and 60+ tumor models have been successfully established, covering major clinical tumor species. 2. Standardized Laboratory Animal Facility Sanyou Bio has rented SPF /Elite animal facility, high-quality laboratory mice and meet animal ethics requirements. 3. High Quality One-stop Service Sanyou Bio is committed to provide you with one-stop high-quality services, from antibody production and purification, tumor cell line construction and customized in vivo efficacy models. 4. Excellent Experience in Project Sanyou Bio is highly experienced and focused on xenograft tumor models for the discovery novel anti-tumor agents (covering Antibodies, Bi/Tri-specific Antibody and Antibody–drug conjugates) with professional knowledge and comprehensive capability. 5. Fast and High-quality Delivery in 56 days Sanyou Bio has a broad range of human xenograft tumor cell lines that are commonly used and provide with customized service to evaluate novel anti-tumor agents in a timely and cost-effective method. Service Features 1. Abundant mouse strains and tumor models 1.1. Abundant mouse strains Sanyou Bio is committed to provide you with alternative strains for in vivo efficiency evaluation, corresponding to mechanisms of action, molecular types, mouse strains. Animal Strain Target Mechanism Molecular Type C57BL/6 Balb/c Nude mice CB-17 SCID NOG NCG NSG Transgenic mice SD/CD mice Immune checkpoint Tumor markers Autoimmunity Tumor microenvironment Angiogenesis Immunoregulation Cytokine ADC Monoclonal antibody Polyspecific antibody Recombinant protein mRNA Small molecules Batch-to batch consistency in cell resources with high stability and low within-group variation. The tumor growth curves of the existing models are as follows: 2. Standardized laboratory animal facility Sanyou has rented SPF /Elite animal facility, high-quality laboratory mice and meet animal ethics requirements. 3. Extensive experience in project research and development 3.1. Extensive experience in target animal models As shown in Table 2, Sanyou Bio has successfully established a series of target candidate molecules, covering dozens of popular targets regarding immunity, autoimmune disorders and metabolism, and we are accumulated in rich project experience for different anti-tumor agents, covering Antibodies, Bi/Tri-specific Antibody, Antibody–drug conjugates and recombinant proteins. Table 2 In-vivo efficiency evaluation with profile Target MOA Solution 1 Solution 2 Reference 1 Reference 2 TIGIT Immune checkpoint hTIGIT-mice A375-PBMC humanized mice Tiragolumab PVRIG Immune checkpoint hPVRIG-mice PBMC humanized mice COM701 TNFR2 Immunity hTNFR2-mice PBMC humanized mice Opi Vi CTLA-4 Immune checkpoint hCTLA-4-mice PBMC humanized mice Ipilimumab EGFR RTKs A431 FaDu Centuximab Panitumumab 4-1BB Immunity CT26 MC38 ABL111 ADG106 CD39 Immunity MOLP-8 TTX030 ROR1 ADC A549 MDA-MB-231 Cirmtuzumab Trop2 ADC A431 MDA-MB-231 Sacituzumab CLDN6 Cancer-associated markers PA-1 CLDN6-OV90 IMAB027 LAG3 Immunity hLAG3-mice PBMC humanized mice Relatlimab HLX26 CD47 Immunity Raji SKOV-3 Magrolimab lemzoparlimab PD-L1 Immune checkpoint hPD-L1-mice PBMC humanized mice Atezolizumab Durvalumab PD-1 Immune checkpoint hPD-1-mice PBMC humanized mice Pembrolizumab Nivolumab VEGF Angiogenesis COLO-205 A431 Bevacizumab Ramucirumab ANGPTL3 Cardiovascular DIOObesity model Evinacumab ALX RTKs A549 MDA-MB-231 Enapotamab DDR1 Immunity CT26 B16-F10 U.Texas CLDN18.2 Cancer-associated markers MC38-hCLDN18.2 HEK293-hCLDN18.2 IMAB362 CD40 Immunity hCD40-mice Selicrelumab HER2 RTKs BT474 N87 Trastuzumab Pertuzumab CD24 Cancer-associated markers HT29 MCF-7 Tel Aviv U CD100 Immunity CT-26 PBMC humanized mice Pamrevlumab TSLP Autoimmune OVA Model Tezepelumab BCMA Cancer-associated markers NCI-H929 MM.1S GSK2857916 Case Stastics 1. 4-1BB: Synergistic CT-26 colon model Synergistic CT26 colon cancer model in Balb/c mice is shown in Fig. 1. The mice were divided into control group and treatment group, and then they were intraperitoneally administered BIW.3 (twice per week for three weeks). Antibody A dosed at 3 MPK showed 95% tumor inhibition, which could significantly inhibit tumor growth. Fig. 1 Tumor growth inhibition 2. HER2: Xenograft N87 gastric cancer model Xenograft N87 gastric cancer model in nude mice is shown in Fig. 2. The mice were divided into control group and treatment group, and then they were intraperitoneally administered BIW.4 (twice per week for four weeks). The results showed that Antibody B dosed at 20 MPK display 85.5% tumor inhibition and Antibody C dosed at 20 MPK has 53% tumor inhibition. Fig. 2 Tumor growth inhibition 3. VEGF: Xenograft COLO-205 colon cancer model Xenograft COLO-205 colon cancer model in nude mice is shown in Fig. 3. The mice were divided into control group and treatment group, and then they were intraperitoneally administered BIW.3 (twice per week for three weeks). The results showed that tumor inhibition of Bevacizumab dosed at 0.95 MPK is as high as 42%. Fig. 3 Tumor growth inhibition 4. CD47: Xenograft Raji lymphoma model Xenograft Raji lymphoma model in NOD/ SCID mice model is shown in Fig. 4. The mice were divided into control group and treatment group, and then they were intravenously administered BIW.3 (twice per week for three weeks). The results showed that magrolimab has dosage-dependent tumor growth inhibition. Tumor inhibition of Magrolimab dosed at 5 MPK is as high as 89.3%, which could significantly inhibit tumor growth, indicating that the model was successfully constructed. Fig. 4 Tumor growth inhibition 5. BCMA: Xenograft MM 1S myeloma model Xenograft MM.1S myeloma model in NOD/ SCID mice is shown in Fig. 5. The mice were divided into control group and treatment group, and then they were intravenously administered BIW.3 (twice per week for three weeks). The results showed tumor inhibition of GSK2857916 dosed at 5 MPK is up to 53%, indicating that the model was successfully constructed. Fig. 5 Tumor growth inhibition 6. CD40: Xenograft Romas lymphoma model Xenograft Romas lymphoma model in Balb/c nude mice is shown in Fig. 6. The mice were divided into control group and treatment group, and then they were intraperitoneally administered BIW.3 (twice per week for three weeks). The results showed that the tumor inhibition of APX005 dosed at 5 MPK is 92.7%. Fig. 6 Tumor growth inhibition 7. MOLP-8 Xenograft myeloma model Xenograft MOLP-8 myeloma model in NCG mice is shown in Fig. 7. The mice were divided into control group and treatment group, and then they were intraperitoneally administered BIW.3 (twice per week for three weeks). The results showed that competitor D could inhibit tumor growth, indicating that the model was successfully constructed. Fig. 7 Tumor growth inhibition 8. CTLA-4: Synergistic MC38 colon cancer model Synergistic MC38 colon cancer in CTLA-4 humanized mice is shown in Fig. 8. The mice were divided into control group and treatment group, and then they were intraperitoneally administered TIW.3 (three times per week for three weeks). The results showed that ipilimumab has a dose-dependent tumor inhibitory effect, and ipilimumab dosed at 2 MPK has tumor inhibition rate of 99%, indicating that the model was successfully constructed. Fig. 8 Tumor growth inhibition 9. TNFR2: Synergistic MC38 colon cancer model Synergistic MC38 colon cancer in in TNFR2 humanized mice is shown in Fig. 9. The mice were divided into control group and treatment group, and then they were intraperitoneally administered BIW.3 (twice per week for three weeks). The results showed that the tumor inhibition rate of OPI is 99.9% at the dose of 15 MPK, indicating that the model was successfully constructed. Fig. 9 Tumor growth inhibition 10. LAG3: Synergistic MC38 colon cancer model Synergistic MC38 colon cancer in LAG3 humanized mice is shown in FIG. 10. The mice were divided into control group and treatment group, and then they were intraperitoneally administered BIW.3 (twice per week for three weeks). The results showed that the tumor inhibition rate of avelumab and relatimab combination is 76.2%, which could significantly inhibit tumor growth, indicating that the model was successfully constructed. Fig. 10 Tumor growth inhibition 11. EGFR: Xenograft A431 head and neck cancer model Xenograft A431 head and neck cancer model in NCG mice is shown in FIG. 11. The mice were divided into control group and treatment group, and then they were intraperitoneally administered BIW.3 (twice a week for consecutive three weeks). The results showed that the tumor inhibition rate of Panitumumab is 84.8% at the dose of 0.7 MPK, which could significantly inhibit tumor growth, indicating that the model was successfully constructed. Fig. 11 Tumor growth inhibition 12. ADC: Xenograft HT-29 colon cancer model Xenograft HT-29 colon cancer model in Balb/c nude mice is shown in Fig. 12. The mice were divided into control group and treatment group, and then they were intravenously administered BIW.4 (twice a week for consecutive four weeks). The results showed that the tumor inhibition rate of competitor E-MMAE was 89.3%, which could significantly inhibit tumor growth, indicating that the model was successfully constructed. Fig. 12 Tumor growth inhibition 13. TriAb: Xenograft N87 gastric cancer model Xenograft N87 gastric cancer model in NCG humanized PBMC mice is shown in Fig. 13. The mice were divided into control group and treatment group, and then they were intravenously administered twice a week for four weeks. The results showed that TriAb has synergistic effect, and the tumor inhibition rate of TriAb was up to 83.4% at the same molar dose, indicating that the model could be used to evaluate the efficacy of tri-specific antibodies. Fig. 13 Tumor growth inhibition 14. PD-L1/ VEGF combination: Xenograft COLO205 colon cancer model Xenograft COLO-205 colon cancer model in NCG humanized PBMC mice is shown in Fig. 14. The mice were divided into control group and treatment group, and then they were intravenously administered twice a week for two weeks. The results showed synergistic effect, and BsAb has the tumor inhibition rate as high as 90.6% at the same molar dose, which showed that the model could be used to evaluate the efficacy of bispecific antibody. Fig. 14 Tumor growth inhibition 15. Biomarkers detection After the in vivo efficacy evaluation of antibody, the liver of mice was taken and tested for alanine aminotransferase (ALT) and aspartate aminotransferase (AST) to evaluate its hepatotoxicity. The results were shown in FIG. 15, the ALT and AST levels of BM was consistent with the patent, indicating that the detection method was reliable. Fig. 15 The levels of ALT and AST in mice liver