Antibody-Drug Conjugate (ADC) | Medicilon Platforms

Solutions:

Medicilon promises careful planning, meticulous execution and delivery of accurate results.
Up until the end of 2022, Medicilon undertook more than 100 major IND application biopharmaceutical projects, including monoclonal antibodies, bispecific antibodies, polyclonal antibodies, ADCs, viral vaccines and fusion proteins. As of the end of 2023, Medicilon has successfully assisted 24 ADC drugs to obtain clinical approval, and more than 20 ADC projects are in development.
Payload (50+): DX8951f, SN-38, DM1, DM4, Exatecan, Dxd, MMAE/MMAF, Tubulysin M, PBD dimer, Seco-Duocarmycin MA, PNU159682, Doxorubicin analog. Linker (200+): Various types of Cleavable and uncleavable, hydrophilic and hydrophobic, and new types of Linker, such as: Gly-Gly-Phe-Gly, VC-PAB, SMCC, N-SMP. Payload+Linker (30+): SN-38-PEG-PAB, DXD-GlyGlyPheGly, DXD-VCPABDM1-SMCC, DM1-SPP, DM4-SPDB, MC-VC-PAB-MMAE/MMAF, MC-Val-Ala-PBD, VC-PAB-Dolastatin 10. Medicilon has completed targets: Her2, Her3, Trop2, Claudin 18.2, CD33, Muc1, FR.
Medicilon has extensive experience in developing and verifying analysis methods for different targets, and can effectively analyze the expression level and accessibility of targets according to the needs, and provide advice for the selection of targets.

Synthesis of ADC Payloads

Medicilon offers our clients a broad range of payload drugs possessing various cell killing mechanisms. We also offer custom synthesis of payload drugs.
Medicilon's compound library has a variety of chemical ADC payloads with different mechanisms of action for customers to choose from. At the same time, ADCs can be customized and synthesized according to the specific needs of customers.
Tubulin inhibitorsDNA damaging agentsImmunomodulators
What are the components of antibody-drug conjugates?
ADC drugs are composed of three parts: antibody, linker, and toxic payload.
Antibody: The antibody is responsible for target engagement, it can be in form of Mab, Fab, Bispecific Ab or nanobody.
Conjugate (Linker):Conjugates the payload to antibodies. The linker connects antibody and payload, and can be either cleavable or non-cleavable in nature. It is key to ADC stability and controls when the payload is released.
Payload:The payload is a highly potent toxin responsible for killing cancer cells.
Medicilon Case:
ADC crosslinking strategy based on cysteine:
Provide 5mg, 50mg and 500mg scale of ADC crosslinking service, timeline: 2-4weeks.Linker-payload: MC-MMAE, MC-MMAF, MC-GGFG-DX8951, MC-SN38 etc.QC methods including SEC, HIC and LC-MS/MS.DAR evaluation through HPLC and LC-MS/MS.

ADC in Vitro Analysis

With more than 300 cancer cell lines, the Medicilon Biological team can offer a wide selection of ADC target protein positive and negative tumor cells. The Medicilon Biological team has extensive experience in cell labeling and FACS-based cell viability analysis to help identify the optimal antibody candidate.
Medicilon Case:
ADC Binding Assay
A: HER2 ADCs (BB-1701 & DS-8201) were incubated with N87 cells and then analyzed through FACS, MFI of PE on secondary antibodies against ADCs were calculated.
B：HER2 protein was immobilized on M5 chip, DS-8201 was serial diluted and injected into the chip, binding affinities of HER2 and DS-8201 was analyzed through Biacore 8K.
FACS-based ADC binding assay (Figure A: HER-2 ADC DS-8202, BB-1701 binding with BT-474 cells which is HER2 highly expressing cells). SPR analysis of ADC with antigen in protein level (Figure B: DS-8201 binding with HER2 protein). Provide Kd, Kon, Koff values for binding. Provide other methods for ADC binding, e.g. ELISA and HTRF
ADC Internalization: Confocal Imaging
OVCAR-3 cells were incubated with FITC-labeled ADC for 24 hours, the cells were incubated with Lyso-Tracker and DAPI and then analyzed through con-focal microscope.
ADC were labeled with fluorophore（e.g. ADC-FITC, ADC-Cy5). Internalization of ADC-FITC were analyzed through con-focal (co-localization of lyso-tracker with ADC-FITC indicating internalization of ADC). Internalization could also be analyzed through LICOR (In cell Western) and FACS.
Cytotoxicity of Payloads or ADC
Payload, naked antibody and ADC were incubated with target cells, cell viability was assessed through CCK-8, CTG and MTT assays.
MV-4-11 cells and PC-3 cells were treated with compound A and stained with PI for FACS-based cell cycle analysis. The data show that compound A dramatically block the cell cycle of MV-4-11 cells but did not affect PC-3 cells significantly.
Apoptosis Analysis
MV-4-11 cells were treated with Compound A and stained with PI/Annexin V for FACS-analysis. The data shown that Compound A promotes the apoptosis of MV-4-11 cells.

Pharmacology Research of ADC

Target antigen binding activityRelated pharmacology of target antigen (e.g.: ADCC，CDC)Mechanism of payloads and metabolites (focus on the difference in pharmacology mechanism ADCs, naked antibodies,payload and metabolites). Effects of the free small molecule compounds after internalization, disassembly and cracking

Pharmacology Evaluation of ADC

Efficacy testing of ADC in vivo is an important pharmacological parameter in ADC research, which can affect the design of clinical trials. Medicilon is committed to providing clients with mature tumor models for evaluating the pharmacodynamics of ADCs in vivo, in an AAALAC-certified environment, completing relevant pharmacodynamic evaluation tests to high standards.
Medicilon has established nearly 300 types of tumor evaluation models in six categories, which can cover most cancer types. ADC drug evaluation models: transgenic, xenograft tumor transplantation, homogeneous tumor transplantation, PDX, humanized tumor transplantation and orthotopic transplantation models are available A variety of experimental animals: rodents (mice/rats, rabbits); non-rodents (beagles, miniature pigs, non-human primates)
Medicilon Case:
In vivo antitumor activity of RC68-based ADCs.
A humanized anti-EGFR monoclonal antibody (named RC68) was purifed and conjugated with MMAE using a MC-VC-PAB or PY-VC-PAB linker. The in vivo antitumor activity of RC68-MC-VC-PAB-MMAE and RC68- PY-VC-PAB-MMAE were assessed by Medicilon.
BALB/c nude mice were implanted subcutaneously with H125 cells and when the solid tumor reached 100-300 mm3 the mice were randomized and treated intravenously with the indicated drug weekly. The effect of each treatment on the growth of tumors was measured by monitoring tumor volumes and their body weights were measured twice per week. At the end of the experiment, the tumors were dissected and photo imaged.

ADC Pharmacokinetics Study

Medcilon provides high quality quantification assays for key parameters in ADC PK study, presenting accurate results.

Analyte	Description	Common analysis methods
Conjugated Antibody	Antibody with minimum of DAR>=1	LBA
Total Antibody	Conjugated, partially unconjugated and fully unconjugated(DAR>=0)	LBA
Small Molecules	Released/free samllmolecule and its metabolities	LC-MS/MS
ADA	Antibodies against antibody of ADC, linker or drug	LBA

Benchmarking-with-global-lab-standard-for-results-with-high-consistency.webp

Benchmarking with global lab standard for results with high consistency. Developing stable and reliable methods for results with high correlation.

ADC Immunogenicity

Immunogenicity is a key parameter when evaluating biologic therapeutics. It could increase the potential risk of adverse effects and reduced ADC efficacy. Medicilon fully understands the complexity of ADA evaluation and offers our clients with comprehensive immunogenicity assays.

ADC Safety assessment

Medicilon offers rigorous and specific safety assessment services strictly following S6 & S9 Regulation of ICH and in compliance with the requirement of the NMPA, FDA, OECD and TGA regulatory agencies.

Single dose/Repeat dose toxicity (With TK)；
Tissue cross-reactivity；
ADA test.

Service Cases

Medicilon assisted Bio-Thera's bispecific neutralizing antibody drug for injection BAT2022 for the treatment of new coronary pneumonia was approved for clinical use