Antibody-Drug Conjugate (ADC) Preclinical DMPK and Bioanalysis Research Strategies

Blogs
Aug 15, 2024

Antibody-drug conjugates (ADCs) are a novel therapeutic that combines highly targeted monoclonal antibodies with cytotoxic small molecule drugs, and have become a rising trend due to their precise and powerful mode of action. ADCs have the characteristics of both small molecule and macromolecular therapeutics, and are often present in vivo as complex and dynamically changing mixtures resulting from biotransformation and DAR changes. These bring unique challenges to ADC DMPK and bioanalytical research. This blog will summarize the marketed ADC drugs and introduce the comprehensive DMPK and bioanalytical strategies for ADC drugs.

Summary of 15 approved ADCs by 2024

As of July 2024, 15 ADC drugs have been approved by regulatory authorities worldwide, covering multiple indications such as hematologic tumors and solid tumors.

ADC drug	Trade name	Maker	Disease indication	Payload/payload class	Payload action	Target	mAb	Linker	DAR	Approval year
Mirvetuximab soravtansine	ELAHERE	ImmunoGen	Ovarian Cancer	Maytansinoid DM4	Microtubule inhibition	FRα	IgG1	Enzyme cleavable	3.5	FDA 2022
Tisotumab vedotin-tftv	Tivdak	Seagen Inc	Cervical cancer	MMAE /auristatin	Microtubule inhibition	Tissue factor	IgG1	Enzyme cleavable	4	FDA 2021
Disitamab vedotin	Aidixi	RemeGen	Gastric carcinoma	MMAE /auristatin	Microtubule inhibition	HER2	IgG1	Enzyme cleavable	4	NMPA 2021
Cetuximab sarotalocan	Akalux	Rakuten Medical	Head and neck cancer	Photosensitizing dye	Damage cell membrane	EGFR	Chimeric IgG1	Non-cleavable	1.3-3.8	PMDA 2021
Loncastuximab tesirine-lpyl	Zynlonta	ADC Therapeutics	Large B-cell lymphoma	SG3199/PBD dimer	DNA damage	CD19	IgG1	Enzyme cleavable	SG3199/PBD dimer	FDA 2021
Belantamab mafodotin-blmf	Blenrep	GlaxoSmithKline (GSK)	Multiple myeloma	MMAF /auristatin	Microtubule inhibition	BCMA	IgG1	Non-cleavable	4	FDA 2020, withdrawn on 22 Nov. 2022
Sacituzumab govitecan	Trodelvy	Immunomedics	Triple-negative breast cancer	SN-38 /camptothecin	TOP1 inhibition	TROP2	IgG1	Acid cleavable	7.6	FDA 2020
Trastuzumab deruxtecan	Enhertu	AstraZeneca/Daiichi Sankyo	HER2-positive breast cancer	DXd /camptothecin	TOP1 inhibition	HER2	IgG1	Enzyme cleavable	8	FDA 2019
Enfortumab vedotin	Padcev	Astellas/Seagen Genetics	Urothelial cancer	MMAE /auristatin	Microtubule inhibition	Nectin4	IgG1	Enzyme cleavable	3.8	FDA 2019
Polatuzumab vedotin-piiq	Polivy	Genentech, Roche	Diffuse large B-cell lymphoma	MMAE /auristatin	Microtubule inhibition	CD79	IgG1	Enzyme cleavable	3.5	FDA 2019
Moxetumomab pasudotox	Lumoxiti	Astrazeneca	Hairy cell leukemia	PE38 (Pseudotox)	Immunotoxin	CD22	IgG1	Cleavable	N/A	FDA 2018
Inotuzumab ozogamicin	Besponsa	Pfizer/Wyeth	Lymphoblastic leukemia	Ozogamicin /calicheamicin	DNA damage	CD22	IgG4	Acid cleavable	6	FDA 2017
Trastuzumab emtansine	Kadcyla	Genentech, Roche	HER2-positive metastatic breast cancer	DM1 /maytansinoid	Microtubule inhibition	HER2	IgG1	Non-cleavable	3.5	FDA 2013
Brentuximab vedotin	Adcetris	Seagen Genetics, Millennium/Takeda	Relapsed HL and relapsed sALCL	MMAE /auristatin	Microtubule inhibition	CD30	IgG1	Enzyme cleavable	4	FDA 2011
Gemtuzumab ozogamicin	Mylotarg	Pfizer/Wyeth	Relapsed acute myelogenous leukemia	Ozogamicin /calicheamicin	DNA damage	CD33	IgG4	Acid cleavable	2–3	FDA 2017; 2000

Table 1. Summary of 15 approved ADCs

DMPK research contents of ADCs

ADCs feature a complex structure and diverse dynamic processes in vivo, and their ADME characteristics are highly correlated with their pharmacological efficacy and toxicity. Carrying out the following DMPK studies for ADC drugs is recommended.

Recommended DMPK Studies	Object	Guidance/white paper description
In-vitro stability	ADC	Before clinical study, need to evaluate the ADC plasma/whole blood stability in human and relevant animal species.
In-vivo PK	Total antibody/conjugated antibody/payload/major metabolites	In-vivo PK generally monitors total antibodies, conjugated antibodies, and free payloads or major metabolites.
Met ID and biotransformation	Linker-payload	For new linker-payloads, it is necessary to understand the major metabolites.
Plasma protein binding	Payload/major metabolites	If the payload or major metabolites is new.
Drug metabolism phenotyping	Payload/major metabolites
Transporters	Payload/major metabolites
Enzyme induction/inhibition	Payload/major metabolites
In-vivo ADME	Payload/major metabolites

Table 2. Recommended DMPK studies for ADC drugs

Integrated bioanalytical strategies for ADC drugs

The absorption, distribution, metabolism, and excretion of ADC drugs in various tissues directly affect the efficacy and safety of ADCs, making their bioanalysis more challenging than other drugs. Not only large molecules (total antibodies and conjugated antibodies) but also small molecule components (free and conjugated payloads) need to be taken into consideration. Bioanalysis of total antibodies, conjugated antibodies, free payloads, and related metabolites is essential throughout all stages, including early discovery/screening, preclinical, and clinical development of ADC drugs. A comprehensive bioanalytical approach is required to properly assess the ADME properties of ADC drugs.

Figure 1. Bioanalytical considerations for ADC drugs

By utilizing multiple analytical platforms and employing a combination of small molecule/large molecule and hybrid analysis methods, suitable analysis strategies can be devised to tackle specific challenges in ADC drug development effectively.

Platforms	Applications	Notes
LBA	Total antibody Conjugated antibody (ADC) Anti-drug antibody (ADA) Neutralizing antibody (Nab)	Bioanalysis of conjugated antibody requires anti-payload antibody
Low-resolution mass spectrometry (triple quadrupole)	Free payload Conjugated payload Payload related metabolites Signature peptide	Often supported by Met ID (metabolite identification) data
High-resolution mass spectrometry	Met ID	Plasma/tumor cell/hepatocyte metabolite identification is performed as needed
Hybrid LBA-LC/(HR)MS	In vitro/in vivo dynamic monitoring of DAR values ADC biotransformation Conjugated antibody (ADC)	Fit for purpose design based on ADC structure; if anti-payload antibody is not available, semi-quantification of ADC
Quantitative whole-body autoradiography QWBA	ADC payload distribution study	Pre-isotope labeling of the payload is required

Table 3. ADC bioanalytical platform and applications

Figure 2. DMPK integrated bioanalytical platform of ADC drugs.jpg

Figure 2. DMPK integrated bioanalytical platform of ADC drugs

In recent years, ADC drugs have gone through multiple iterations to improve efficacy and safety, and have been updated to fourth-generation ADC drugs. ADC drugs' stability, safety, and efficacy require a combination of multiple biological analysis platforms. As more innovative conjugation strategies, antibody scaffolds, and novel warheads are used in next-generation ADCs, bring more challenges to ADC bioanalysis.

Summary

With extensive experience in ADC drug bioanalysis, WuXi AppTec DMPK has established integrated bioanalytical platforms (LBA, LC-MS, LC-HRMS, hybrid LBA/LC-(HR)MS, and QWBA), developed and validated bioanalytical methods to evaluate the components of ADCs, to provide customers with one-stop ADC bioanalysis services. We have supported hundreds of ADC projects, including dozens of ADC IND applications.

DMPK Studies	Pre-clinical Discovery	Pre-clinical Development
DMPK Studies	Lead optimization	ADME Characterization and IND application
ADME	Plasma/serum/blood stability of ADC Identification of metabolites related to payload released by ADC in vitro	Microsomal/hepatocyte metabolite identification of payload Plasma protein binding of payload ADME study of radiolabeled payload in animals In vivo metabolite identification of ADC
DDI	Study on the permeability and efflux transport of payload	Study on inhibition and induction of P450 enzyme by payload Metabolic enzyme phenotyping of payload Study on the inhibition of drug transporter by payload
PK	PK study of ADC or payload in rodents Screening PKPD study of ADC	PK studies of ADC or payload in non-rodents PKPD study of ADC

Table 4. DMPK studies to support ADC preclinical discovery and development

Committed to accelerating drug discovery and development, we offer a full range of discovery screening, preclinical development, and clinical drug metabolism and pharmacokinetic (DMPK) platforms and services. With research facilities in the United States (New Jersey) and China (Shanghai, Suzhou, Nanjing, and Nantong), 1,000+ scientists, and over fifteen years of experience in Investigational New Drug (IND) application, our DMPK team at WuXi AppTec are serving 1,600+ global clients, and have successfully supported 1,500+ IND applications.

Talk to a WuXi AppTec expert today to get the support you need to achieve your drug development goals.

Authors: Maotian Zhou, Huijuan Qian

Reference

1. Ruan DY, Wu HX, Meng Q, Xu RH. Development of antibody-drug conjugates in cancer: Overview and prospects. Cancer Commun (Lond). 2024 Jan;44(1):3-22. doi: 10.1002/cac2.12517. Epub 2023 Dec 30. PMID: 38159059; PMCID: PMC10794012.

Receive DMPK email updates

Talk to an Expert

Related Services and Platforms

Stay Connected

Keep up with the latest news and insights.

Email address*

* Please check the filled content
First name*

* Please check the filled content
Last name*

* Please check the filled content
Company*

* Please check the filled content

By clicking submit, you consent to WuXi AppTec DMPK collecting and processing the information you provide for our internal purposes, in accordance with our privacy policy.

* Please agree to the Privacy Policy

Submit