novel humanized monoclonal antibodies to pd-l1€¦ · novel humanized monoclonal antibodies to...

Abeome Corporation of 17

Novel Humanized Monoclonal Antibodies to PD-L1

Using Abeome’s novel transgenic mouse antibody discovery platform, we have rapidly

obtained antibodies of high affinity and neutralizing potency against human PD-L1 (Programmed

Death Ligand 1, CD274). B-cells expressing affinity-matured anti-PD-L1 surface antibody were

directly selected, and recombinant chimeric antibodies were immediately cloned and screened for

PD-L1 binding and neutralization. With no additional optimization or affinity maturation steps, we

generated humanized lead candidates that have comparable, and potentially superior, affinity and

neutralization potency to atezolizumab, a humanized anti-PD-L1 mAb recently approved for the

treatment of urothelial and non-small cell lung cancer (NSCLC). The further in vivo evaluation of

these lead molecules should support a valid clinical development path. For licensing information,

please contact us:

Contacts Marty Simonetti [email protected] Kirby Alton [email protected] Rick Shimkets

[email protected]

Laboratories 111 Riverbend Road Athens, GA 30605 T- (706)542-7889

mailto:[email protected]




Contents:

I. PD-L1 Background

II. Abeome Antibody Discovery Platform

III. Immunization With Human PD-L1 Extracellular Domain IV. Harvest and Single Antigen-Positive Cell Sorting

V. High-Throughput Screening of PD-L1 Antibodies

VI. Identification and Characterization of Leads with PD-L1 Neutralizing Function

VII. Humanization Approach and Technology

VIII. Lead Characterization: Expression, Potency & Affinity

IX. PD-L1 Neutralization Primary T Cell Assay

X. Summary of Lead Discovery

XI. Therapeutic Product Development

I. PD-L1 (Programmed Death Ligand 1; B7-H1; CD274)

Research in the last two decades has established the concept that the PD-1/PD-L1

pathway acts to inhibit T cell activation and can be exploited by tumors to escape immune attack in

the tumor microenvironment [1]. Activation of PD-1 on the surface of T cells by engagement with

PD-L1 inhibits human T cell responses in vitro [2], and many in vivo studies have revealed that the

PD-1/PD-L1 pathway plays a major role in the suppression of T cell responses [3,4]. The PD-L1

protein is abundantly expressed on the cell surface in various human cancers, as indicated by

immunohistochemistry [5]. In contrast, normal human tissues seldom express PD-L1 protein on

their cell surface, with the exception of tonsil, placenta, and a small fraction of macrophage-like

cells in lung and liver [5,6]. Recent clinical success disrupting the PD-1/PD-L1 pathway has shown

the power of immunotherapy and brought about a sea-change in the treatment of cancer. The use

of combination immunotherapy may provide improved outcomes in patients over single-agent

checkpoint blockade. Such combinations may likely involve a PD-1/PD-L1 pathway blocker as its

backbone due to the remarkable therapeutic index of this class of agents [7].

II. Abeome Antibody Discovery Platform: AbeoMouseTM

We have developed a novel transgenic mouse system (AbeoMouseTM) allowing for the

direct selection of antigen-specific B-cells, paired with single-cell antibody gene cloning and screening. The AbeoMouseTM produces a 45-fold increase in surface immunoglobulin (Ig) positive antibody secreting cells and an accelerated immune response. Abeome’s screening platform allows 1,000 times more affinity matured monoclonal antibodies to be isolated from a single AbeoMouseTM than by conventional technology. In contrast to other current antibody technologies, this platform allows for the enrichment and rapid cloning of specific, high-affinity chimeric antibodies against a target of interest. With this modular system, cloned variable regions (V-regions) may be swapped between multiple human Ig isotypes for empirical comparison of stability, affinity and functional potency, or to suit the specific therapeutic modality or effector function.

Specifically, the transgenic AbeoMouseTM has been engineered to constitutively express multiple genes, including the Igα/Igβ B-cell receptor proteins, resulting in a hyper immune response and surface antibody expression during all stages of B cell differentiation (Fig.1). This enables the


selection and sorting of antigen specific B-cells producing the most affinity matured antibodies, and this technology platform has been applied to obtain antibodies against a diverse set of antigens, including but not limited to whole cells, peptides, glycoproteins, viral envelope proteins and mouse proteins, typically producing chimeric leads with low picomolar dissociation constants.

FIGURE 1. The transgenic AbeoMouseTM platform. A novel antibody discovery platform that generates mature B cells with high surface IgG expression, allowing for the direct selection and cloning of antigen-specific B cells

III. Immunization With Human PD-L1 Extracellular Domain

Six AbeoMiceTM 7-8 weeks of age were pre-bled to obtain baseline serum antibody levels

and then immunized subcutaneously with 40 g of recombinant human PD-L1 extracellular domain (N-terminal segment Met 1-Thr 239 with a C-terminal polyhistidine tag; Sino Biological Inc.) in

proprietary adjuvant. Booster injections with 20 g of protein were given at days 14 and 28. Blood samples were taken and serum titers determined at days 21 and 35, resulting in extremely high titers, and mice were harvested at approximately 45 days after initial immunization (Fig. 2, Table 1).


FIGURE 2. Immunization timeline for generating anti-PD-L1 antibodies

Table 1. Immunization and titer summary of mice immunized with PD-L1.

Antigen Proprietary adj-initial injection

1060-1 PDL-1

Boost # 1 Bleed # 1 Titer # 1 Boost # 2 Bleed # 2 Titer # 2

Female ms # 963 5/27/2015 SC 6/10/2015 6/17/2015 2,048K 6/24/2015 7/2/2015 8192K

Female ms # 964 5/27/2015 SC 6/10/2015 6/17/2015 128K 6/24/2015 7/2/2015 8192K

Male ms # 3341 10/09/2015 SC 10/23/2015 10/30/2015 >8,192K 11/6/2015 11/10/2015 > 8,192K

Male ms # 3342 10/09/2015 SC 10/23/2015 10/30/2015 >8,192K 11/6/2015 11/12/2015 > 8,192K

Male ms # 3352 10/09/2015 SC 10/23/2015 10/30/2015 >8,192K 11/6/2015 11/13/2015 8,192K

Male ms # 3353 10/09/2015 SC 10/23/2015 10/30/2015 8.192K 11/6/2015 11/13/2015 8.192K

IV. Harvest and Single Antigen-Positive Cell Sorting (Mouse #963)

As a representative example of this campaign, mouse #963 yielded 26 lymphoid tissue samples and bone marrow (Figure 3) which were harvested, processed, and pooled into a single suspension of lymphoid cells (6.6 x 108 cells). Erythrocytes were lysed using ammonium chloride and resulting lymphocytes were depleted of cells expressing IgM antibodies by immuno-magnetic separation.


2 x 108 IgM depleted cells were labeled for FACS (fluorescent activated cell sorting).

Specifically, cells were first incubated with purified rat anti-mouse CD16/CD32 (mouse Fc

receptor block; BD Pharmingen) to block non-specific binding to mouse Fc receptors. Antigen

specific antibody was detected with ifluor-488 labeled recombinant human PD-L1. Surface

antibody was stained with alexa-fluor 647-GAMA-IgG. Antigen-positive cells were isolated as

single cells by FACS as illustrated in Fig. 4 (red boxed cell population). Cells expressing surface

antibody reactive to both human PD-L1 (ifluor-488) and GAMA-IgG (alexa-fluor 647) were singly

deposited into wells of a 96 well plate. A total of 1920 Ag+/Ig+ clones were sorted into wells from

this mouse.

FIGURE 4. Single cell FACS sorting of lymphocytes expressing antibody to human PD-L1 The direct selection of cells surface-expressing anti-PD-L1 antibodies is accomplished by staining harvested B cells with fluor-labeled recombinant human PD-L1 and a polyclonal antibody recognizing mouse surface immunoglobulin. Double-positive cells (red box) are sorted at 1 cell/well into 96 well plates for RT-PCR and cloning.

FIGURE 3. Lymphoid organs harvested from mouse #963 and processed into a single cell suspension for FACS (fluorescent activated cell sorting). Abeome’s transgenic mice have an enhanced immune response, greatly enlarged lymphoid organs (left), and typically an order of magnitude increase in the number of antibody producing plasmacytes.

Surface IgG

PD

-L1


V. High-Throughput Screening of PD-L1 Antibodies

After cell sorting, twenty-seven 96-well plates of single B cells, comprising >2,000 antibodies, were subjected to nested RT-PCR using heavy and light chain variable region specific primer sets. Amplified V-regions were then fused with mammalian expression promoters and human Fc chains (IgG4 and kappa constant regions) by overlap PCR, generating transcriptionally-active PCR products. These individual paired heavy and light chain PCR products were transfected into HEK293 cells to generate supernatants of secreted chimeric antibodies and screened for binding to PD-L1 by ELISA. A representative set of screening data is shown (Fig. 5), which identified many antibodies positive for PD-L1 binding. In total, we isolated and cloned more than 60 chimeric monoclonal antibodies (mAbs) that showed strong binding to human PD-L1.


FIGURE 5. Representative ELISA screening data for anti-PD-L1 chimeric antibodies. High-throughput screening of >1,000 novel antibodies revealed many with strong binding to human PD-L1. Values are expressed as human PD-L1 binding signal normalized to total IgG expression level.

VI. Identification and Characterization of Leads with PD-L1 Neutralizing Function

In order to identify potent, neutralizing antibody leads to human PD-L1, we utilized a PD-L1 neutralization reporter assay. Briefly, this two-cell system consists of a transgenic Jurkat line expressing a NFAT luciferase reporter and PD-1, and a CHO line expressing a TCR activator and human PD-L1 (Fig. 6). When co-cultured under normal conditions, the TCR activation signal in the Jurkat reporter line is blocked by engagement of PD-1 with PD-L1 on the cell surface, resulting in low luciferase activity. Neutralization of the PD-1/PD-L1 interaction by an antibody relieves this blockade and results in increased luciferase activity. The assay may be used to measure antibody potency by dose-dependent increase in luciferase activity.


Figure 6. PD-L1 Neutralization Reporter Assay System

As a first step, the PD-L1 neutralization reporter assay was used to screen our set of novel PD-L1 antibodies at a single concentration (Figure 7) to identify all chimeric mAbs with neutralizing function. From this reporter assay screen, we identified 11 chimeric mAbs that resulted in strong activation, indicating neutralization of PD-L1. To further characterize these lead chimeric mAbs, we sequenced the mouse V-regions from our set of mAbs positive for PD-L1 binding. Interestingly, from the approximately 60 mAbs isolated from our screens, we classified eight sequence families, with a single sequence family representing all mAbs that exhibited PD-1/PD-L1 neutralizing activity in cell assay screens.


Figure 7. Representative results from PD-L1 neutralization reporter assay screens. The figure shows the increased luciferase activity produced by 11 of the cloned antibodies. In the initial screen, approximately 60 antibodies exhibiting strong binding to PD-L1 were incubated with PD-L1 expressing-CHO TCR activator cells at a single concentration. This first cellular screen identified several candidate antibodies that showed neutralization of the PD-L1/PD-1 interaction. (add control description)

To identify the lead candidate chimeric mAbs with the highest potency in neutralizing PD-L1, we then performed dose titrations of the lead neutralizing mAbs in the reporter assay to determine IC50 values, and the activity of our lead mAbs was compared to a humanized clinical lead anti-PD-L1 mAb, atezolizumab. Atezolizumab (Tecentriq®) was developed by Genentech/Roche and was recently approved for the treatment of urothelial cancer and NSCLC. As shown in Figure 8, we identified a lead chimeric mAb, ABM101, with comparable activity to

atezolizumab. Specifically, ABM101 had an IC50 of 0.042 g/mL (290 pM) compared to 0.039

g/mL (269pM) for atezolizumab. In addition, we measured the affinity to human PD-L1 of ABM101 versus the clinical lead by surface plasmon resonance (SPR, Biacore). As we had shown for potency, we observed that ABM101 had a comparable affinity to atezolizumab in our experiments (Table 2).


Figure 8. Neutralization of PD-L1 in a cell assay by lead chimeric mAb ABM101 is comparable to a clinical control, atezolizumab.

Table 2. Potency (IC50) and affinity (SPR) of lead chimeric mAb ABM101 compared to humanized clinical control antibody, atezolizumab.

VII. Humanization Approach and Technology

Abeome’s high-throughput monoclonal antibody discovery process begins with the cloning of mouse variable regions recovered from single B cells into proprietary human constant region-containing vectors, thus generating chimeric antibodies, which are tested for desirable properties. Ideally, multiple functional antibodies are selected to move forward into humanization, because in some cases it is not possible to design successful human framework grafts, while in other cases it is possible to obtain humanized antibodies with properties superior to the parent chimera.

Abeome’s approach to humanization comprises the generation of multiple heavy and light chain grafts, and pairwise testing to determine whether functional grafts can be immediately obtained. Structural models of each mouse fAb are generated and compared to the designed grafted fAb structure. As needed, back mutations are made in the grafted constructs in order to attempt to bring the structural models into alignment and maximize retention of parental affinity. Potential post-translational modification sites and other manufacturing challenges, such as non-canonical or unpaired cysteine residues and N-glycosylation sites are identified and engineered


away. If additional affinity is required, mutagenesis of CDRs is undertaken using proprietary methods.

VIII. Humanized Lead Characterization: Expression, Potency & Affinity

Using our system of CDR grafting on human frameworks and proprietary expression vectors, we cloned a total of 6 heavy chain and 4 light chain humanized variations from the original ABM101 parent sequences. Many of the humanized antibody derivatives were subsequently cloned in IgG1 (N298A) and IgG4 (S228P) configurations. To characterize the humanized leads, we transiently expressed them in the Expi293 and ExpiCHO cell systems (Life Technologies) and measured the expression, potency and affinity of the purified mAbs. Examples of the potency and high affinity of the Abeome lead humanized candidates can be seen in Figures 9 and 10, respectively. Table 3 represents a summary of the potency and affinity data of the humanized leads. We did not see significant binding of any of the ABM101 family humanized mAbs to recombinant mouse PD-L1. In addition, we did not see any measurable binding of hABM101.11 to other human B7 family proteins (VISTA, B7-H3; data not shown).

Figure 9. Comparison of lead candidates A) hABM101.9 and B) hABM101.11 expressed as IgG1 and IgG4 isotypes to atezolizumab activity in the PD-L1 neutralization reporter cell assay.


Figure 10. Multicycle Kinetics of ABM101.11 IgG1 and ABM101.9 IgG4 vs recombinant human (rhu) and cynomolgus (rcyno) monkey PD-L1. Affinity of anti-PD-L1 mAbs was

determined by surface plasmon resonance (SPR) on a Biacore T100. Anti-PD-L1 mAbs (25 g/mL) were captured onto a Protein A sensorchip and multicycle kinetics vs recombinant human or cynomolgus monkey PD-L1 (40 nM-2.5 nM) was determined. The antibody-PD-L1 complex was allowed to dissociate for 20 minutes before regenerating the surface with 10 mM glycine-HCL, pH 1.5. Data for ABM101.11 IgG1 and ABM101.9 IgG4 is shown.


Table 3. Summary of humanized ABM101 family of anti-PD-L1 antibody chains, expression, potency (IC50), and affinity measurements. IC50 values are averages from multiple reporter cell assays, and affinity was measured by SPR binding to human and cynomolgus monkey recombinant PD-L1 extracellular domain. *KD values for atezolizumab were obtained from published reports [10, 11].

IX. PD-L1 Neutralization Primary T Cell Assay

To validate the function of our PD-L1 mAbs in primary human cells, we tested three of our

lead humanized mAbs in a primary human T cell assay. Studies by investigators in the field have

shown that PD-L1 extracellular domain coated on beads can inhibit the anti-CD3 proliferative

stimulation of and IFN secretion from T cells [8,9]. The neutralization function of antibodies may

be tested in this assay by blocking the negative effects of PD-L1. Briefly, magnetic beads were

coated with anti-CD3 antibody (OKT3), and then additionally coated with human IgG-Fc control

protein or human PD-L1-Fc protein. The coated beads were mixed with various concentrations of

purified anti-PD-L1 humanized mAbs, and the bead-antibody mixes were incubated with purified

human CD4+ T cells. After 5 days, cell proliferation (CellTiter Glo, Promega) and IFN secretion

(ELISA, R&D Systems) was measured, and antibody activity was assessed by neutralization of the

PD-L1 mediated decrease in cellular proliferation or IFN protein levels from control (IgG-Fc coated

beads). PD-L1 coated beads alone reduced proliferation by over 30% compared to the control IgG

beads, and all antibodies tested completely blocked the inhibition of proliferation by PD-L1 (Fig.

11A). Interestingly, the ABM101.11 and ABM101.9 mAbs show an increase in proliferation above

the control beads. For IFN secretion, we saw a modest decrease with PD-L1 beads, but this

ABM# Isotype

mouse

ABM

parent antigen H chain L chain

HEK

Expression

(mg/L)

CHO

Expression

(mg/L)

IC50

(g/mL)

KD (pM) by

SPR

(HUMAN)

KD (pM) by

SPR

(CYNO)

ABM101 IgG4 PD-L1 ABM101H ABM101L 19.7 0.042 647 pM

ABM101 IgG1 PD-L1 ABM101H ABM101L 13.9 0.19

hABM101.1.1 IgG4 ABM101 PD-L1 ABM101.1H ABM101.1L 4.7 0.52

hABM101.2.1 IgG4 ABM101 PD-L1 ABM101.2H ABM101.2L 2.18 n/a

hABM101.1.2 IgG4 ABM101 PD-L1 ABM101.1H ABM101.2L 26.5 0.62

hABM101.2.2 IgG4 ABM101 PD-L1 ABM101.2H ABM101.1L 3.77 n/a

hABM101.1.3 IgG4 ABM101 PD-L1 ABM101.1H ABM101L 7.47 0.18

hABM101.2.3 IgG4 ABM101 PD-L1 ABM101.2H ABM101L 0.82 0.28

hABM101.1.4 IgG4 ABM101 PD-L1 ABM101H ABM101.1L 4.43 0.05

hABM101.2.4 IgG4 ABM101 PD-L1 ABM101H ABM101.2L 10.9 0.26

hABM101.3 IgG4 ABM101 PD-L1 ABM101.3H ABM-101.1L 29.21 0.044

hABM101.3.2 IgG4 ABM101 PD-L1 101.3H (10390-C5) ABM101.1L 112.9 n/a

hABM101.3.3 IgG4 ABM101 PD-L1 101.3H (10390-E9) ABM101.1L 150.4 n/a

hABM101.3.4 IgG4 ABM101 PD-L1 101.3H (10391-E1) ABM101.1L 127.3 n/a

hABM101.3.5 IgG4 ABM101 PD-L1 101.3H (10391-F1) ABM101.1L 2.26 0.21

hABM101.4 IgG4 ABM101 PDL1 ABM101.4H ABM101.1L 14.79 0.046

hABM101.6 IgG4 ABM101 PDL1 ABM101.5H ABM101.3L 5.29 0.029

hABM101.7 IgG1 ABM101 PD-L1 ABM101.6H ABM101.4L 14.5 0.046

hABM101.7 IgG4 ABM101 PD-L1 ABM101.6H ABM101.4L 30.64 0.036 31 pM 39 pM




hABM101.9 IgG4 ABM101 PD-L1 ABM101.4H ABM101.4L 8.31 5.15 0.035 152 pM 171 pM


hABM101.11 IgG4 ABM101 PD-L1 ABM101.5H ABM101.4L 23.9 0.018 349 pM

atezolizumab IgG1 PD-L1 10.3 0.025 160-400pM*


decrease was abolished by treatment with all antibodies tested (Fig. 11B), and as with proliferation,

we observed an increase in signal beyond the control beads. This data provides further evidence

that the novel humanized lead anti-PD-L1 mAbs are effective at neutralizing the PD-1 pathway in

normal human T cells, and that these novel mAbs perform in a similar fashion to atezolizumab.

Figure 11. Primary Human CD4+ T Cell Assays of ABM101-family antibodies. After 5 days of

growth, the percent control viable cells (A) and IFN secretion (B) was measured on primary

human CD4+ T cells incubated with control or PD-L1 coated beads. Humanized lead anti-PD-L1


lead mAbs show effective neutralization PD-L1 blockade and show similar activity when compared

to atezolizumab (ATZ). The dashed lines represent the baseline against which the antibody

treatment samples are being compared.

X. Summary of Lead Discovery

Using our novel transgenic mouse antibody discovery platform, we have shown that we can

rapidly obtain chimeric antibodies of high affinity and neutralizing potency against human PD-L1.

B-cells expressing affinity-matured anti-PD-L1 surface antibody were directly selected, and

recombinant chimeric antibodies were immediately cloned and screened for PD-L1 binding and

neutralization. With no additional optimization or affinity maturation steps, we generated

humanized lead candidates that have comparable, if not superior, affinity and neutralization

potency to atezolizumab, a humanized anti-PD-L1 mAb recently approved for the treatment of

urothelial cancer and metastatic NSCLC. The selection of a final lead antibody and isotype will be

achievable upon pharmacokinetic and pharmacodynamic evaluation, which will further

demonstrate their potential clinical utility.

XI. Therapeutic Product Development

Pre-clinical studies in mice have shown that administration of either anti-PD-1 or anti-PD-L1

antibodies inhibit the growth of both liquid (myeloma) and solid tumors and the metastatic spread

of melanoma and colon cancer cells [12]. Other studies have shown that blockade of the PD-1

pathway in vitro or in vivo can rescue the effector function of T cells, promoting their proliferation

and survival, and leading to increased inflammatory cytokine production and cytolytic activity

against tumor cells.[13,14]. Abeome’s lead PD-L1 antibody candidates do not bind mouse PD-L1

and therefore will require the use of human cancer models during preclinical evaluation. However,

many “humanized” preclinical mouse models of cancer are available from various CRO’s, and the

anti-tumor efficacy and preliminary ADME of these leads can be analyzed in these models.

Indeed, durvalumab (AstraZeneca) serves as an example case of an anti-PD-L1 antibody that

does not cross-react to the mouse ortholog (Table 4), and preclinical development of this molecule

was carried out in mice with human cancer cell lines.

The three major anti-PD-L1 biologics in development (Table 4) have each been granted

Breakthrough Therapy Designation and priority review by the US FDA. Atezolizumab (Genentech)

was approved in May 2016 for treatment of urothelial cancer and in October 2016 for NSCLC. A

Biological License Application (BLA) was filed in December 2016 for durvalumab (AstraZeneca)

seeking approval for the treatment of urothelial cancer and a BLA was filed for avelumab (Pfizer) in

November 2016 for the treatment of Merkel Cell Carcinoma. Upon the successful conclusion of

pharmacokinetic, pharmacological, and toxicology testing, clinical development of Abeome’s novel

anti-PD-L1 antibody could follow a similar path.


Table 4. Summary of Current and Approved Anti-PD-L1 Antibody Agents

References:

1. Chen L, Han X. Anti–PD-1/PD-L1 therapy of human cancer: past, present, and future. J

Clin Inv. 2015;125(9):3384-3391.

2. Dong H, Zhu G, Tamada K, Chen L. B7-H1, a third member of the B7 family, co-stimulates

T-cell proliferation and interleukin-10 secretion. Nat Med. 1999;5(12):1365–1369.

3. Chen L. Co-inhibitory molecules of the B7-CD28 family in the control of T-cell immunity. Nat

Rev Immunol. 2004;4(5):336–347.

4. Zou W, Chen L. Inhibitory B7-family molecules in the tumour microenvironment. Nat Rev

Immunol. 2008;8(6):467–477.

5. Dong H, et al. Tumor-associated B7-H1 promotes T-cell apoptosis: a potential mechanism

of immune evasion. Nat Med. 2002;8(8):793–800.

6. Petroff MG, Chen L, Phillips TA, Hunt JS. B7 family molecules: novel immunomodulators at

the maternal-fetal interface. Placenta. 2002;23(suppl A):S95–S101.

7. Atkins M. Immunotherapy combinations with checkpoint inhibitors in metastatic melanoma:

Current approaches and future directions. Semin Oncol. 2015 Dec;42 Suppl 3:S12-9.

8. Bennett F, Luxenberg D, Ling V, Wang IM, Marquette K, Lowe D, Khan N, Veldman G,

Jacobs KA, Valge-Archer VE, Collins M, Carreno BM. Program death-1 engagement upon

TCR activation has distinct effects on costimulation and cytokine-driven proliferation:

attenuation of ICOS, IL-4, and IL-21, but not CD28, IL-7, and IL-15 responses. J Immunol.

2003 Jan 15;170(2):711-8.

9. Freeman GJ, Long AJ, Iwai Y, Bourque K, Chernova T, Nishimura H, Fitz LJ, Malenkovich

N, Okazaki T, Byrne MC, Horton HF, Fouser L, Carter L, Ling V, Bowman MR, Carreno BM,

Collins M, Wood CR, Honjo T. Engagement of the PD-1 immunoinhibitory receptor by a

novel B7 family member leads to negative regulation of lymphocyte activation. J Exp Med.

2000 Oct 2;192(7):1027-34.

10. Irving B, Chiu H, Maecker H, Mariathasan S, Lehar SM, Wu Y, Cheung J. Anti-PD-L1

antibodies, compositions and articles of manufacture. United States patent US 8,217,149.

2012 Jul 10.

11. Herbst RS, et al. Predictive correlates of response to the anti-PD-L1 antibody MPDL3280A

in cancer patients. Nature. 2014 Nov 27;515(7528):563-7.

12. Baksh K, Weber J. Immune checkpoint protein inhibition for cancer: preclinical justification

for CTLA-4 and PD-1 blockade and new combinations. Semin Oncol 2015;42:363–77.

Non-proprietary

Name Isotype ADCC

Mouse

PD-L1

Binding Pre-Clinical Model Testing Phase I Phase II Phase III

FDA Approved

Indications

durvalumab

IgG1-kappa

(modified) no -

Xenograft with human T cell

grafted mice (NOD/SCID) and

HPAC (pancreatic), A375

(melanoma) human cancer

Numerous Cancers,

including liquid and

solid tumors

Numerous Cancers,


solid tumors

Head & Neck, NSCLC,

Breast, Bladder,

Squamous Cell Lung

Carcinoma

atezolizumab

IgG1-kappa

(N297A) no +

chronic LCMV mouse model;

MC38Ova (colon) syngeneic

mouse cancer model

Numerous Cancers,


solid tumors

Lymphomas, RCC,

Bladder, Kidney,

NSCLC

Non-Squamous-

NSCLC, NSCLC, RCC,

Breast,

Bladder/Urothelial

Cancer

Urothelial Cancer,

NSCLC

avelumab IgG1-lambda yes +

MC38 (colon), C1498 (leukemia),

PANC02 (pancreatic) syngeneic

mouse cancer models

Solid tumors, renal

cancer (RCC), NSCLC,

Hodgkins Lymphoma,

Merkel Cell Carcinoma

Merkel Cell

Carcinoma, NSCLC

NSCLC, RCC, Gastric

Cancer, Ovarian

Cancer, Urothelial

Cancer


13. Peng W, Liu C, Xu C, et al. PD-1 blockade enhances T-cell migration to tumors by

elevating IFN-gamma inducible chemokines. Cancer Res 2012;72:5209–18.

14. Zhou Q, Xiao H, Liu Y, et al. Blockade of programmed death-1 pathway rescues the

effector function of tumor-infiltrating T cells and enhances the antitumor efficacy of

lentivector immunization. J Immunol 2010;185:5082–92.

novel humanized monoclonal antibodies to pd-l1€¦ · novel humanized monoclonal antibodies to...

Documents