Does sacituzumab-govitecan act as a conventional antibody drug conjugate (ADC), a prodrug of SN-38 or both?

On April 22, 2020, the Food and Drug Administration granted accelerated approval to sacituzumab-govitecan (Trodelvy) for adult patients with metastatic triple-negative breast cancer (TNBC) who received at least two prior therapies for metastatic disease (1). Sacituzumab-govitecan is an antibody drug conjugate (ADC) of a humanized anti-Trop2 monoclonal antibody (mAb), RS7, linked to an average of 7.6 molecules of SN-38—the active metabolite of irinotecan and a potent inhibitor of Topoisomerase 1 (Topo1) (2). By chemically connecting the drug and mAb the goal was to deliver and release SN-38 to tumor cells abundant in Trop2. In the Phase III ASCENT study sacituzumab-govitecan demonstrated clinical benefit in patients with metastatic TNBC irrespective of Trop2 expression, albeit with greater efficacy in patients with a medium or high Trop2 score (3,4). In contrast, although sacituzumab-govitecan was efficacious in metastatic small cell lung cancer (mSCLC), progression-free and overall survival showed no clear relationship to Trop2 expression (5). Hence, there is an enigma as to how the anti-Trop2 ADC can be effective in cancers that either have or have not the Trop2 antigen. A knowledge of the mechanism of action of sacituzumab-govitecan in Trop2 low or absent tumors would facilitate further development of SN-38-based drugs or ADCs targeting Trop2, and we present hypotheses here that explains this effect.

It was anticipated that if sacituzumab-govitecan internalized in tumor cells, a protease site on the linker would be cleaved by lysosomal enzymes to release SN-38 intracellularly. However, the internalization of sacituzumab-govitecan may not be very efficient. In early efforts to establish Trop2 targeting, tumor uptake of the carrier mAb 131I-RS7 was only ~7% to 16% of the initial dose/gm in a Trop2 TNBC xenograft—only ~2-fold higher than a control 131I-mAb (6); by comparison, Trastuzumab shows an uptake of ~40% of the initial dose/gm in a HER2-positive tumor (7). However, in sacituzumab-govitecan, the linker attaching the monoclonal antibody to SN-38 also contains a hydrolysable carbonate moiety that has a cleavage half-life of only ~18 hours in neutral pH or sera—the “weakest link in the chain”. It has been suggested that the hydrolytically labile linker allows time-dependent extracellular release of free drug in the tumor microenvironment so it can affect adjacent cells by a bystander effect (2).

The rapid spontaneous linker hydrolysis in sacituzumab-govitecan releases a very large amount of the SN-38 cargo systemically (8), much more than with other ADCs—which are generally designed to avert spontaneous drug release—or that can be accounted for by a targeted mechanism with limited target capacity. Thus, the question should be asked as to whether the antitumor effects of sacituzumab-govitecan are due to a conventional ADC mechanism, a bystander effect, systemically released SN-38, or a combination thereof.

The below shows the pharmacokinetic parameters of the SN-38 generated from sacituzumab-govitecan and from irinotecan—the prototypical SN-38 prodrug—in the human. Although the mechanisms of SN-38 generation are quite different, the exposure, or AUC, of the SN-38 released from sacituzumab-govitecan over a three-week cycle is over 15-fold higher than that from irinotecan at their maximally tolerated doses; also, the time over the target (TOT) concentration of ~10 nM SN-38 needed to inhibit Topo1 is significantly longer with sacituzumab-govitecan. Since the much lower levels of SN-38 generated from irinotecan have significant therapeutic and toxicity effects, it can be confidently concluded that much higher systemic levels of free SN-38 released from sacituzumab-govitecan must have equal or greater pharmacological effects. This could explain the efficacy of sacituzumab-govitecan observed in low Trop2 score mSCLC (5) and mTNBC (4).

Table 1

Pharmacokinetic parameters of SN-38 generated from sacituzumab-govitecan and irinotecanA

Drug	SN-38 Cmax, nM	SN-38 t1/2, h	TOT3 WkB Days >10 nM	SN-38 AUC3 Wk, μM × h
Sacituzumab-govitecan 10 mg/kg, Q Wk, 2/3 Wks	320	18	7.5	20C
Irinotecan, 340 mg/m2 Q3 Wk	140	21	4.4	1.2

A, the Cmax, t1/2 and single-dose AUC values were obtained from the FDA label information for sacituzumab-govitecan and CPT-11 (Camptosar); B, average time SN-38 is over 10 nM target concentration over 3 Wk; C, the AUC3 Wk of SN-38 released from sacituzumab-govitecan over 3 Wk was calculated as twice the reported AUC168 hr. Wk, week; AUC, area under the curve; TOT, time on target.

Since sacituzumab-govitecan is so effective in TNBC, one can rightly ask the importance of understanding its mechanism. First, it would certainly be important to know whether Trop2 is indeed a suitable target to either encourage or dissuade work on Trop2-targeted ADCs with different payloads. As noted above, sacituzumab-govitecan is very active in small cell lung cancer but its efficacy is unrelated to Trop2 expression (5); also, PF-06664178, a potent Trop2-targeted ADC linked to a protease-cleavable auristatin has not fared well in early clinical trials (9). Second, whether sacituzumab-govitecan is or is not a target-directed ADC or prodrug would influence the stability of linkers chosen for similar therapeutics; DS-1062a and SKB264 are Trop2-targeting ADCs in early trials that also have Topo1 poison payloads—exatecan and belotecan, respectively—attached by protease-labile linkers that would not be released systemically and require intracellular delivery and payload release. Third, if maintenance of a high systemic concentration of free SN-38 is essential for efficacy of sacituzumab-govitecan it may limit the use of certain drug combinations. Currently, sacituzumab-govitecan is administered IV on days 1 and 8 of a 21-day cycle, so there is a near-continuous high systemic exposure of SN-38 with a minimal drug-free interval. PARP inhibitors are highly synergistic with Topo1 inhibitors such as SN-38, but the synergy exists for toxicities as well as efficacy. For example, the current sacituzumab-govitecan dosing schedule does not provide a sufficient SN-38-free interval in which a PARP inhibitor could be safely administered—as for example by using a gap schedule approach for combinations of Topo1 inhibitors and DNA damage response inhibitors (10)—hence, the use of a sacituzumab-govitecan-PARP inhibitor combination may not be feasible. Indeed, a recent small trial of sacituzumab-govitecan and rucaparib indicated efficacy of the combination, but at the expense of significant early grade 3/4 neutropenia (11). Finally, if the high AUC of free SN-38 is a major driver of sacituzumab-govitecan efficacy, a properly designed long-acting prodrug of SN-38 could achieve that AUC, as well as a lower Cmax to lower systemic toxicity and a prolonged half-life to increase time over target; moreover, use of a prodrug would not be confined to tumors that have Trop2. Hence, comparing the efficacy of a long-acting non-targeted SN-38 prodrug to sacituzumab-govitecan at doses that provide equal exposure may resolve to what extent sacituzumab-govitecan acts as a SN-38 prodrug versus a targeted ADC.

Conclusions

Pharmacokinetic and biomarker data, together with considerations of its rapidly hydrolyzed linker, suggests that sacituzumab-govitecan might act as an SN-38-prodrug instead of or in addition to a conventional ADC.

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