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Quantitative Investigation of Pharmacokinetics of Antibody-Based Therapeutics in the Central Nervous System.

紀錄類型:	書目-電子資源 : Monograph/item
正題名/作者:	Quantitative Investigation of Pharmacokinetics of Antibody-Based Therapeutics in the Central Nervous System./
作者:	Chang, Hsueh-Yuan.
出版者:	Ann Arbor : ProQuest Dissertations & Theses, : 2021,
面頁冊數:	385 p.
附註:	Source: Dissertations Abstracts International, Volume: 82-09, Section: B.
Contained By:	Dissertations Abstracts International82-09B.
標題:	Pharmaceutical sciences. -
電子資源:	https://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=28316913
ISBN:	9798582510291

Quantitative Investigation of Pharmacokinetics of Antibody-Based Therapeutics in the Central Nervous System.
Chang, Hsueh-Yuan.

Quantitative Investigation of Pharmacokinetics of Antibody-Based Therapeutics in the Central Nervous System. - Ann Arbor : ProQuest Dissertations & Theses, 2021 - 385 p.

Source: Dissertations Abstracts International, Volume: 82-09, Section: B.

Thesis (Ph.D.)--State University of New York at Buffalo, 2021.

This item must not be sold to any third party vendors.

There has been a great interest in developing antibody-based therapeutics to treat central nervous system (CNS) disorders. Although much progress has been demonstrated using preclinical models, success has not been proven in clinical trials. This disappointment may be due to our limited understanding of the monoclonal antibody (mAb) entrance and clearance in the CNS. We also lack reliable methodologies to accurately investigate the PK of mAbs at the site-of-action in the clinic. Without such measurement, it is challenging to confirm if the mAb concentrations at the site-of-action reach a desired therapeutic level. In the absence of knowledge about drug concentrations at the site-of-action, it also becomes challenging to design an optimal dosing regimen for the patients in the clinic. In this dissertation, a large-pore microdialysis technique has been developed and used to understand the disposition of antibody-based therapeutics in different regions of the brain, using rats as animal models. Besides, a physiologically-based pharmacokinetic (PBPK) model has been developed, which is capable of characterizing the PK of antibody-based therapeutics in different regions of the brain following systemic administration, intracranial (IC) injection, intracerebroventricular (ICV) injection, and intrathecal (IT) injection.In the first section of this dissertation, we have focused on developing a push-pull large-pore microdialysis system for immunoglobulin G (IgG) (150 kDa) and investigated the PK of nonbinding mAbs in different regions of the brain following intravenous injection. We validated the coordinates for implantation of microdialysis guide cannulas into three selected brain regions and developed sensitive and selective sandwich ELISA methods to quantify very low human IgG1 (Trastuzumab) concentrations present in relatively abundant endogenous rat IgGs in CNS. In order to accommodate the limited sample volume from microdialysis, the ELISA was further modified using a 384-well plate format, which requires a relatively low sample volume (30 uL). Our results reveal the heterogeneous distribution of endogenous rat IgG and exogenous human IgG in the CNS, which may question the accuracy of current practice to use the cerebrospinal fluid (CSF) as a surrogate for the site-of-action mAb concentration in the clinical trials. A relatively higher mAb concentration in CSF at cisterna magna (CM) than CSF at lateral ventricles (LV) was observed in our studies. This finding is consistent with a reported protein concentration gradient in the CSF circulatory system. In this section, we have also augmented a whole-body platform PBPK model for antibodies to include a more physiological brain compartment. The brain compartment in this model accounts for CSF circulation, perivascular pathway, and newly discovered lymphatic pathway. Only three parameters were estimated to develop the model for rats, and the model was able to reasonably capture mAb PK data in rat plasma, brain, interstitial fluid (ISF), and CSF. The translation of the model to other species was validated in mice, monkeys, and humans by only changing the physiological parameters.In the second section of this dissertation, we further examined the effect of FcRn binding on mAb disposition in the brain by comparing the PK of FcRn-nonbinding Trastuzumab and wild-type Trastuzumab in rat brain following systemic administration. We introduced IHH mutation in the Fc region of Trastuzumab to develop the FcRn nonbinding version. This mutation has been reported to disable the FcRn binding while having no significant impact on Fcγ receptor binding. The purity and molecular weight of IHH-Trastuzumab was checked using SDS-PAGE. We found that the ratio of brain-to-plasma exposure for IHH-Trastuzumab was significantly decreased compared to Trastuzumab. This finding suggests that FcRn may play a role in the FcRn-mediated influx of antibodies in the brain following systemic administration. We also observed that the terminal slope of IHH-Trastuzumab in the brain ISF was slower than plasma, which suggests that FcRn may also play a role in the efflux of mAb from the brain. As such, our observations suggest FcRn may play a role in both influx and efflux of mAb across the brain.In the third section of this dissertation, we have evaluated the effect of receptor-mediated transcytosis (RMT) on mAb disposition in the brain. We have investigated brain PK of four different anti-transferrin receptor (TfR) mAbs with different binding affinities for rat TfR using the microdialysis system following systemic administration. We confirmed that the free mAb concentration might be increased at the brain ISF and whole-brain homogenate for anti-TfR mAbs with an optimal binding affinity. We also found the accumulation of high-affinity anti-TfR mAbs within brain endothelial cells, which is consistent with previous findings that suggest limited distribution of high-affinity anti-TfR mAbs within the brain due to accumulation inside the vascular endothelial cells. We also confirmed the bell-shaped relationship between TfR binding affinity and antibody exposure in the brain. In addition, our previously developed brain PBPK model for antibodies was further expanded in this section to account for RMT of antibodies across the blood-brain barrier (BBB) and the blood-CSF barrier (BCSFB). The expression and turnover parameters for TfR were estimated using rat and mouse PK data. The model was then translated to other species and validated using literature data. The model was able to reasonably capture many of the published anti-TfR antibody PK data in the brain.In the fourth section of this dissertation, we have investigated the effect of size on protein therapeutic disposition in different regions of the rat brain using a large-pore microdialysis system, following systemic administration. F(ab)2, F(ab), and single-chain variable fragments (scFv) of Trastuzumab were produced and purified, and administered in the animal via intravenous injection or continuous intravenous infusion. We found a bell-shaped relationship between protein size and ISF exposure. An increasing CSF exposure with a decreasing molecular weight was also observed. We also observed that the region with the highest tissue-to-plasma exposure ratio differs based on the size of the protein. As such, our observations suggest that there could be an optimal size of protein therapeutics to accomplish maximum selective exposure of the drug at the desired location within the CNS.In summary, the body of work presented in this dissertation investigates brain disposition of nonbinding antibodies, antibody fragments, and antibodies demonstrating RMT, using a novel large-pore microdialysis system developed by us. A translational brain PBPK model to characterize protein therapeutic disposition in the brain is also developed, which can predict plasma PK, whole-brain PK, ISF PK, and CSF PK of proteins following intravenous, IC, ICV, and IT administration. The results presented in this dissertation could serve as a good starting point to establish a quantitative understanding of antibody and protein therapeutic disposition in different regions of the brain, including the site-of-action. The brain PBPK model developed here may serve as a quantitative modeling tool to support the discovery, preclinical-to-clinical translation, and clinical development of novel protein therapeutics to treat various CNS disorders.

ISBN: 9798582510291Subjects--Topical Terms:

3173021
Pharmaceutical sciences.
Subjects--Index Terms:

Antibody disposition in the brain

Quantitative Investigation of Pharmacokinetics of Antibody-Based Therapeutics in the Central Nervous System.
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520 $a There has been a great interest in developing antibody-based therapeutics to treat central nervous system (CNS) disorders. Although much progress has been demonstrated using preclinical models, success has not been proven in clinical trials. This disappointment may be due to our limited understanding of the monoclonal antibody (mAb) entrance and clearance in the CNS. We also lack reliable methodologies to accurately investigate the PK of mAbs at the site-of-action in the clinic. Without such measurement, it is challenging to confirm if the mAb concentrations at the site-of-action reach a desired therapeutic level. In the absence of knowledge about drug concentrations at the site-of-action, it also becomes challenging to design an optimal dosing regimen for the patients in the clinic. In this dissertation, a large-pore microdialysis technique has been developed and used to understand the disposition of antibody-based therapeutics in different regions of the brain, using rats as animal models. Besides, a physiologically-based pharmacokinetic (PBPK) model has been developed, which is capable of characterizing the PK of antibody-based therapeutics in different regions of the brain following systemic administration, intracranial (IC) injection, intracerebroventricular (ICV) injection, and intrathecal (IT) injection.In the first section of this dissertation, we have focused on developing a push-pull large-pore microdialysis system for immunoglobulin G (IgG) (150 kDa) and investigated the PK of nonbinding mAbs in different regions of the brain following intravenous injection. We validated the coordinates for implantation of microdialysis guide cannulas into three selected brain regions and developed sensitive and selective sandwich ELISA methods to quantify very low human IgG1 (Trastuzumab) concentrations present in relatively abundant endogenous rat IgGs in CNS. In order to accommodate the limited sample volume from microdialysis, the ELISA was further modified using a 384-well plate format, which requires a relatively low sample volume (30 uL). Our results reveal the heterogeneous distribution of endogenous rat IgG and exogenous human IgG in the CNS, which may question the accuracy of current practice to use the cerebrospinal fluid (CSF) as a surrogate for the site-of-action mAb concentration in the clinical trials. A relatively higher mAb concentration in CSF at cisterna magna (CM) than CSF at lateral ventricles (LV) was observed in our studies. This finding is consistent with a reported protein concentration gradient in the CSF circulatory system. In this section, we have also augmented a whole-body platform PBPK model for antibodies to include a more physiological brain compartment. The brain compartment in this model accounts for CSF circulation, perivascular pathway, and newly discovered lymphatic pathway. Only three parameters were estimated to develop the model for rats, and the model was able to reasonably capture mAb PK data in rat plasma, brain, interstitial fluid (ISF), and CSF. The translation of the model to other species was validated in mice, monkeys, and humans by only changing the physiological parameters.In the second section of this dissertation, we further examined the effect of FcRn binding on mAb disposition in the brain by comparing the PK of FcRn-nonbinding Trastuzumab and wild-type Trastuzumab in rat brain following systemic administration. We introduced IHH mutation in the Fc region of Trastuzumab to develop the FcRn nonbinding version. This mutation has been reported to disable the FcRn binding while having no significant impact on Fcγ receptor binding. The purity and molecular weight of IHH-Trastuzumab was checked using SDS-PAGE. We found that the ratio of brain-to-plasma exposure for IHH-Trastuzumab was significantly decreased compared to Trastuzumab. This finding suggests that FcRn may play a role in the FcRn-mediated influx of antibodies in the brain following systemic administration. We also observed that the terminal slope of IHH-Trastuzumab in the brain ISF was slower than plasma, which suggests that FcRn may also play a role in the efflux of mAb from the brain. As such, our observations suggest FcRn may play a role in both influx and efflux of mAb across the brain.In the third section of this dissertation, we have evaluated the effect of receptor-mediated transcytosis (RMT) on mAb disposition in the brain. We have investigated brain PK of four different anti-transferrin receptor (TfR) mAbs with different binding affinities for rat TfR using the microdialysis system following systemic administration. We confirmed that the free mAb concentration might be increased at the brain ISF and whole-brain homogenate for anti-TfR mAbs with an optimal binding affinity. We also found the accumulation of high-affinity anti-TfR mAbs within brain endothelial cells, which is consistent with previous findings that suggest limited distribution of high-affinity anti-TfR mAbs within the brain due to accumulation inside the vascular endothelial cells. We also confirmed the bell-shaped relationship between TfR binding affinity and antibody exposure in the brain. In addition, our previously developed brain PBPK model for antibodies was further expanded in this section to account for RMT of antibodies across the blood-brain barrier (BBB) and the blood-CSF barrier (BCSFB). The expression and turnover parameters for TfR were estimated using rat and mouse PK data. The model was then translated to other species and validated using literature data. The model was able to reasonably capture many of the published anti-TfR antibody PK data in the brain.In the fourth section of this dissertation, we have investigated the effect of size on protein therapeutic disposition in different regions of the rat brain using a large-pore microdialysis system, following systemic administration. F(ab)2, F(ab), and single-chain variable fragments (scFv) of Trastuzumab were produced and purified, and administered in the animal via intravenous injection or continuous intravenous infusion. We found a bell-shaped relationship between protein size and ISF exposure. An increasing CSF exposure with a decreasing molecular weight was also observed. We also observed that the region with the highest tissue-to-plasma exposure ratio differs based on the size of the protein. As such, our observations suggest that there could be an optimal size of protein therapeutics to accomplish maximum selective exposure of the drug at the desired location within the CNS.In summary, the body of work presented in this dissertation investigates brain disposition of nonbinding antibodies, antibody fragments, and antibodies demonstrating RMT, using a novel large-pore microdialysis system developed by us. A translational brain PBPK model to characterize protein therapeutic disposition in the brain is also developed, which can predict plasma PK, whole-brain PK, ISF PK, and CSF PK of proteins following intravenous, IC, ICV, and IT administration. The results presented in this dissertation could serve as a good starting point to establish a quantitative understanding of antibody and protein therapeutic disposition in different regions of the brain, including the site-of-action. The brain PBPK model developed here may serve as a quantitative modeling tool to support the discovery, preclinical-to-clinical translation, and clinical development of novel protein therapeutics to treat various CNS disorders.
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