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Effect of the Size of Protein Therapeutics on Brain Pharmacokinetics Following Systematic Administration

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A Correction to this article was published on 24 May 2022

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Abstract

Here, we have investigated the effect of size of protein therapeutics on brain pharmacokinetics (PK) following systemic administration in rats. All tested proteins were derived from trastuzumab that do not bind to any targets in rats. PK data generated with F(ab)2 (100 kDa), Fab (50 kDa), and scFv (27 kDa) fragments of trastuzumab, along with published PK data for FcRn non-binding and wild-type trastuzumab (150 kDa), were used to establish a relationship between the protein size and brain exposure. A large-pore microdialysis system was used to measure the PK of proteins in the plasma, the interstitial fluid (ISF) at the striatum (ST), and the cerebrospinal fluid (CSF) at the lateral ventricle (LV) and cisterna magna (CM). Concentrations of all the proteins in plasma, brain homogenate, ISF, and CSF were measured using ELISA. When evaluating the effect of protein size in the absence of FcRn binding, we found a bell-shaped relationship between the size and ISF/plasma AUC ratio, where 100 kDa F(ab)2 demonstrated the highest exposure. A similar bell-shaped relationship was observed for the brain homogenate/plasma AUC ratio, with a peak at 50 kDa. The CSF/plasma AUC ratio at LV increased monotonously with a decrease in the size of proteins. We observed that the exposure of protein therapeutics in different regions of the brain could be significantly different and there could be optimal sizes of protein therapeutics to accomplish maximum/selective exposure in selected brain regions following systemic administration.

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Abbreviations

BBB:

Blood-brain barrier

BCSFB:

Blood-cerebrospinal fluid barrier

CHO:

Chinese Hamster Ovary

CM:

Cisterna magna

CNS:

Central nervous system

CSF:

Cerebrospinal fluid

CSF(CM):

CSF at the CM

CSF(LV):

CSF at the LV

ELISA:

Enzyme-linked immunosorbent assay

IgG:

Immunoglobulin G

ISF:

Interstitial fluid

ISF(ST):

ISF at ST

IV:

Intravenous

KD:

Dissociation constant

LV:

Lateral ventricles

mAbs:

Monoclonal antibodies

PD:

Pharmacodynamic

PK:

Pharmacokinetics

RMT:

Receptor-mediated transcytosis

sdAb:

Single-domain antibody

ST:

Striatum

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Acknowledgements

We thank Dr. Kristin Hill and Dr. Vivian Rodriguez-Cruz for their advice on in vivo catheterization surgery. We also thank Dr. Lena Medina (Instech infusion) for the advice on the blood collection. Finally, we thank Dr. Chunxia Qiao for the advice on the scFv plasmid and the pcDNA5_FRT plasmid.

Funding

This research was funded by the grant from the Center for Protein Therapeutics (CPT) at the University at Buffalo. DKS is also supported by National Institute of General Medical Sciences grant [GM114179], National Institute of Allergy and Infectious Diseases grant [AI138195], and National Cancer Institute grants [R01CA246785 and R01CA256928].

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, School of Pharmacy and Pharmaceutical Sciences, The State University of New York at Buffalo, 455 Pharmacy Building, Buffalo, NY, 14214-8033, USA

    Hsueh-Yuan Chang, Shengjia Wu, Yingyi Li, Leiming Guo, Yuelin Li & Dhaval K. Shah

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  1. Hsueh-Yuan Chang
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  2. Shengjia Wu
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  3. Yingyi Li
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  4. Leiming Guo
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  5. Yuelin Li
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  6. Dhaval K. Shah
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Contributions

Conceptualization: Dhaval K. Shah, Hsueh-Yuan Chang, Shengjia Wu.

ScFv expression in CHO cells: Leiming Guo.

Protein Purification: Hsueh-Yuan Chang, Yingyi Li, Leiming Guo.

In vivo investigation: Hsueh-Yuan Chang, Shengjia Wu, Yuelin Li.

Corresponding authors

Correspondence to Hsueh-Yuan Chang or Dhaval K. Shah.

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Chang, HY., Wu, S., Li, Y. et al. Effect of the Size of Protein Therapeutics on Brain Pharmacokinetics Following Systematic Administration. AAPS J 24, 62 (2022). https://doi.org/10.1208/s12248-022-00701-5

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