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. 2021 Mar 23;10(6):1323.
doi: 10.3390/jcm10061323.

Preclinical Characterization of the Distribution, Catabolism, and Elimination of a Polatuzumab Vedotin-Piiq (POLIVY®) Antibody-Drug Conjugate in Sprague Dawley Rats

Victor Yip  1 , M Violet Lee  2 , Ola M Saad  2 , Shuguang Ma  3 , S Cyrus Khojasteh  3 , Ben-Quan Shen  1
Affiliations

Preclinical Characterization of the Distribution, Catabolism, and Elimination of a Polatuzumab Vedotin-Piiq (POLIVY®) Antibody-Drug Conjugate in Sprague Dawley Rats

Victor Yip et al. J Clin Med. .

Abstract

Polatuzumab vedotin (or POLIVY®), an antibody-drug conjugate (ADC) composed of a polatuzumab monoclonal antibody conjugated to monomethyl auristatin E (MMAE) via a cleavable dipeptide linker, has been approved by the United States Food and Drug Administration (FDA) for the treatment of diffuse large B-cell lymphoma (DLBCL). To support the clinical development of polatuzumab vedotin, we characterized the distribution, catabolism/metabolism, and elimination properties of polatuzumab vedotin and its unconjugated MMAE payload in Sprague Dawley rats. Several radiolabeled probes were developed to track the fate of different components of the ADC, with 125I and 111In used to label the antibody component and 3H to label the MMAE payload of the ADC. Following a single intravenous administration of the radiolabeled probes into normal or bile-duct cannulated rats, blood, various tissues, and excreta samples were collected over 7-14 days post-dose and analyzed for radioactivity and to characterize the metabolites/catabolites. The plasma radioactivity of polatuzumab vedotin showed a biphasic elimination profile similar to that of unconjugated polatuzumab but different from unconjugated radiolabeled MMAE, which had a fast clearance. The vast majority of the radiolabeled MMAE in plasma remained associated with antibodies, with a minor fraction as free MMAE and MMAE-containing catabolites. Similar to unconjugated mAb, polatuzumab vedotin showed a nonspecific distribution to multiple highly perfused organs, including the lungs, heart, liver, spleen, and kidneys, where the ADC underwent catabolism to release MMAE and other MMAE-containing catabolites. Both polatuzumab vedotin and unconjugated MMAE were mainly eliminated through the biliary fecal route (>90%) and a small fraction (<10%) was eliminated through renal excretion in the form of catabolites/metabolites, among which, MMAE was identified as the major species, along with several other minor species. These studies provided significant insight into ADC's absorption, distribution, metabolism, and elimination (ADME) properties, which supports the clinical development of POLIVY.

Keywords: antibody–drug conjugate (ADC); distribution, catabolism, and elimination (DME); mass balance; monomethyl auristatin E (MMAE); polatuzumab vedotin (POLIVY).

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Conflict of interest statement

The authors are Genentech employees at the time of conducting these studies and declare no conflict of interest.

Figures

Figure 1
Figure 1
Structure of [3H]-monomethyl auristatin E (MMAE)-polatuzumab vedotin (drug-to-antibody ratio (DAR) of 3.4) and the location of the radiolabeled 3H on the MMAE.
Figure 2
Figure 2
The systemic pharmacokinetics (PK) and tissue biodistribution of unconjugated [3H]-MMAE following a single intravenous (IV) administration. (A) Radioactivity of [3H]-MMAE (as a percentage of the injected dose) from the whole blood, plasma, and cell pellets in rats up to 144 h post administration. A six- to eight-fold higher level of radioactivity was observed throughout the study in the cell pellets isolated from blood compared to the plasma, suggesting a strong partition to red blood cells. (B) Radioactivity of [3H]-MMAE from blood and multiple tissues isolated from rats at various time points post IV administration. The data are represented as the mean ± standard deviation (SD). n = 3 per time point. S. Intestine: small intestine; L. Intestine: large intestine.
Figure 3
Figure 3
The route of elimination of unconjugated [3H]-MMAE in rats following a single IV administration. The total 3H radioactivity in feces and urine samples was collected through the study, while bile samples were collected in a separate set of bile-duct-cannulated rats. The data are represented as the mean ± SD. n = 6 per time point up to 24 h (including 24 h), and n = 3 per time point beyond 24 h to the end of the study.
Figure 4
Figure 4
Structure of MMAE and its metabolites identified, with the possible biotransformations in the rat bile samples dosed with [3H]-MMAE.
Figure 5
Figure 5
Profiles of the blood radioactivity levels vs. time of radiolabeled polatuzumab vedotin following a single IV administration. (A) Radioactivity of [125I]-polatuzumab vedotin (as a percentage of the injected dose) comparing with [125I]-polatuzumab antibodies in plasma in rats at tracer (≈3 ug/kg) and tracer plus 10 mg/kg dose levels up to 14 days post administration. The data are represented as the mean ± SD. n = 3 per time point. (B) Radioactivity of [3H]-MMAE-polatuzumab vedotin (as a percentage of the injected dose) from the whole blood, plasma, and cell pellets in rats up to 14 days post administration. The data are represented as the mean ± SD. n = 3 per time point. (C) Plasma from the rats dosed with [3H]-MMAE-polatuzumab vedotin were further processed using organic solvent precipitation, which showed that most of the radioactivity was in the precipitable fraction, suggesting that the MMAE payload was still conjugated to the antibodies in the plasma.
Figure 5
Figure 5
Profiles of the blood radioactivity levels vs. time of radiolabeled polatuzumab vedotin following a single IV administration. (A) Radioactivity of [125I]-polatuzumab vedotin (as a percentage of the injected dose) comparing with [125I]-polatuzumab antibodies in plasma in rats at tracer (≈3 ug/kg) and tracer plus 10 mg/kg dose levels up to 14 days post administration. The data are represented as the mean ± SD. n = 3 per time point. (B) Radioactivity of [3H]-MMAE-polatuzumab vedotin (as a percentage of the injected dose) from the whole blood, plasma, and cell pellets in rats up to 14 days post administration. The data are represented as the mean ± SD. n = 3 per time point. (C) Plasma from the rats dosed with [3H]-MMAE-polatuzumab vedotin were further processed using organic solvent precipitation, which showed that most of the radioactivity was in the precipitable fraction, suggesting that the MMAE payload was still conjugated to the antibodies in the plasma.
Figure 6
Figure 6
The tissue distribution of radiolabeled polatuzumab vedotin and unconjugated polatuzumab antibody after a single dose via IV administration in rats. (A) The tissue and blood radioactivity from intact (125I radioactivity—solid bar) and catabolized (subtract 125I radioactivity from 111In radioactivity—open bar) polatuzumab vedotin dosed at the tracer (≈3 μg/kg) level up to 14 days post administration. (B) The tissues and blood radioactivity from intact (125I radioactivity—solid bar) and catabolized (111In subtracted with 125I radioactivity—open bar) unconjugated polatuzumab antibody dosed at the tracer (≈3–4 μg/kg) level up to 14 days post administration. (C) The tissues and plasma radioactivity from conjugated (3H precipitated radioactivity—solid bar) and deconjugated (3H soluble radioactivity—open bar) [3H]-MMAE-polatuzumab vedotin dosed at 10 mg/kg up to 14 days post administration. A higher distribution of polatuzumab vedotin was observed in highly perfused tissues, such as the liver, spleen, and kidneys, and resulted in higher catabolism. The data are represented as the mean ± SD (except in (C) where only the mean is reported). n = 3 per time point. S. Intestine: small intestine; L. Intestine: large intestine; B. Marrows: bone marrows; A. gland: adrenal glands; DRG: dorsal root ganglions; Sc. Nerve: sciatic nerve.
Figure 6
Figure 6
The tissue distribution of radiolabeled polatuzumab vedotin and unconjugated polatuzumab antibody after a single dose via IV administration in rats. (A) The tissue and blood radioactivity from intact (125I radioactivity—solid bar) and catabolized (subtract 125I radioactivity from 111In radioactivity—open bar) polatuzumab vedotin dosed at the tracer (≈3 μg/kg) level up to 14 days post administration. (B) The tissues and blood radioactivity from intact (125I radioactivity—solid bar) and catabolized (111In subtracted with 125I radioactivity—open bar) unconjugated polatuzumab antibody dosed at the tracer (≈3–4 μg/kg) level up to 14 days post administration. (C) The tissues and plasma radioactivity from conjugated (3H precipitated radioactivity—solid bar) and deconjugated (3H soluble radioactivity—open bar) [3H]-MMAE-polatuzumab vedotin dosed at 10 mg/kg up to 14 days post administration. A higher distribution of polatuzumab vedotin was observed in highly perfused tissues, such as the liver, spleen, and kidneys, and resulted in higher catabolism. The data are represented as the mean ± SD (except in (C) where only the mean is reported). n = 3 per time point. S. Intestine: small intestine; L. Intestine: large intestine; B. Marrows: bone marrows; A. gland: adrenal glands; DRG: dorsal root ganglions; Sc. Nerve: sciatic nerve.
Figure 7
Figure 7
The route of elimination of [3H]-MMAE-polatuzumab vedotin in rats following a single IV administration. The total 3H radioactivity in the feces and urine samples collected through the study. Bile samples were collected in a separate set of bile-duct-cannulated rats. The acetonitrile (ACN) precipitation of urine and feces samples showed that the majority (>80%) of radioactivity was in the ACN-soluble fraction, suggesting that [3H]-MMAE-polatuzumab vedotin was eliminated mainly as [3H]-MMAE or [3H]-containing small molecule catabolites. Similarly, the acetonitrile precipitation of bile samples showed >95% of the radioactivity in the soluble fraction. The data are represented as the mean ± SD. For the urine and feces data, n = 6 per time point up to 7 days (including at day 7), n = 3 per time point from beyond 7 days to the end of the study. n = 6 per time point for the bile samples.
Figure 8
Figure 8
Structure of the catabolites identified, with the possible biotransformations in rat bile samples dosed with [3H]-MMAE-polatuzumab vedotin.
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