JP2023514795A - Methods and compositions for treating cancer - Google Patents

Abstract

The present application provides compositions, methods, and kits for treating cancer, including bladder cancer, such as luminal bladder cancer, using FGFR3 inhibitors in combination with checkpoint inhibitors. do. In some embodiments, the cancer expresses wild-type FGFR3. The FGFR3 inhibitor can be an antagonistic FGFR3 inhibitor, such as an antagonistic FGFR3 antibody. The checkpoint inhibitor can be a PD1 inhibitor (PD1 or PD1 ligand (PD-L1) antibody, such as an antagonistic PD1 or PD-L1 antibody such as).

Description

PRIORITY CLAIM This application is based on U.S. Provisional Patent Application No. 62/812,929 filed March 1, 2019, U.S. Provisional Patent Application No. 62/856,216 filed June 3, 2019, and U.S. Provisional Patent Application No. 62/856,216 No. 62/907,504, filed Sep. 27, 2004, the disclosures of which are hereby incorporated by reference in their entirety, including drawings.

SEQUENCE LISTING This disclosure contains a Sequence Listing which has been submitted in ASCII format via EFS-Web and is incorporated by reference in its entirety. Its ASCII copy (created on March 3, 2020) has the name SequenceListing. txt and is 10 kilobytes in size.

Background Urothelial cell carcinoma (UCC)--also known as transitional cell carcinoma (TCC)--occurs in the urinary system (ie, kidney, bladder, and adnexa) and is the most common type of bladder cancer. , which accounts for 90% of all bladder tumors (Eble 2004). It is the fifth most common cancer in the United States (US) (Costantini 2011) and the fourth most common cancer in Europe (Jemal 2011); an estimated 74,690 new cases and 15 , 580 deaths occurred in the United States in 2014 (American Cancer Society 2018), and an estimated 136,000 new cases and 49,000 deaths occurred in Europe in 2009 (Bellmund 2009). Clinical and pathological studies have identified two variants of urothelial cell carcinoma (UCB) of the bladder, which arise through distinct mechanisms, a low-grade papillary variant and an aggressive tumor variant (Wu 2005). ; Vallot 2010). The low-grade papillary variant accounts for 80% of all UCB and results from urothelial hyperplasia. The 5-year survival rate for this tumor type is greater than 90% when treated with surgery and intravesical immunotherapy (American Cancer Society 2018). The aggressive tumor variants represent 20% of UCB and have a poor prognosis. Cisplatin-based chemotherapy with either dose dense M-VAC (Sternberg 2001) or gemcitabine cisplatin (von der Maase 2000) remains the standard treatment for aggressive UCC. Despite an initial response rate of the order of 50-70%, this cancer typically progresses rapidly, with a median survival of about 13-15 months (von der Maase 2000; Siefker-Radtke 2002). New treatments for UCC are needed.

SUMMARY In certain embodiments and methods of treating cancer in a subject in need thereof, including bladder cancer and upper urothelial cancer, such as metastatic urothelial cancer (mUC) Provided herein are embodiments and methods comprising administering a therapeutically effective amount of an FGFR3 inhibitor and a therapeutically effective amount of a checkpoint inhibitor (eg, a PD1 inhibitor). In certain embodiments, the FGFR3 inhibitor binds FGFR3. In other embodiments, the FGFR3 inhibitor binds a ligand of FGFR3, eg, FGF1, FGF2, or FGF9. In certain embodiments, the FGFR3 inhibitor is an antagonistic FGFR3 antibody, and in certain of these embodiments, the antagonistic FGFR3 antibody comprises a CDR-H1 comprising SEQ ID NO: 1, SEQ ID NO: 2, CDR-H3 with SEQ ID NO:3, heavy chain variable region with SEQ ID NO:7, heavy chain with SEQ ID NO:9, CDR-L1 with SEQ ID NO:4, CDR- with SEQ ID NO:5 L2, CDR-L3 comprising SEQ ID NO:6, a light chain variable region comprising SEQ ID NO:8, and one or more of a light chain comprising the amino acid sequence set forth in SEQ ID NO:10. In certain of these embodiments, the FGFR3 antagonistic antibody is bofatamab. In other embodiments, the antagonistic FGFR3 antibody is selected from the group consisting of PRO-001 and IMC-D11. In certain embodiments, the FGFR3 inhibitor is a small molecule pan-FGFR inhibitor, and in certain of these embodiments, the pan-FGFR inhibitor is infigatinib, AZD4547, LY2874455, pemigatinib, BGJ398 , rogaratinib, PRN1371, Debio 1347, ARQ 087, and JNJ-42756493. In certain embodiments, the PD1 inhibitor binds PD1. In other embodiments, the PD1 inhibitor binds a ligand of PD1, eg, PDL1 or PDL2. In certain embodiments, the PD1 inhibitor is an antagonistic PD1 antibody, and in certain of these embodiments, the antagonistic PD1 antibody is nivolumab, pembrolizumab, CT-011, MEDI-0680 , and RMP1-14. In other embodiments, the PD1 inhibitor is an antagonistic PD1 ligand antibody, and in certain of these embodiments, the antagonistic PD1 ligand antibody is MEDI-4736, RG7446, BMS-936559, MSB0010718C, and MPDL3280A.

In certain embodiments, provided herein are compositions comprising an FGFR3 inhibitor and a checkpoint inhibitor (eg, a PD1 inhibitor). In certain of these embodiments, the composition is a pharmaceutical composition, and in certain embodiments, the composition comprises one or more pharmaceutically acceptable carriers. including. In certain embodiments, the FGFR3 inhibitor binds FGFR3. In other embodiments, the FGFR3 inhibitor binds a ligand of FGFR3, eg, FGF1, FGF2, or FGF9. In certain embodiments, the FGFR3 inhibitor is an antagonistic FGFR3 antibody, and in certain of these embodiments, the antagonistic FGFR3 antibody comprises a CDR-H1 comprising SEQ ID NO: 1, SEQ ID NO: 2, CDR-H3 with SEQ ID NO:3, heavy chain variable region with SEQ ID NO:7, heavy chain with SEQ ID NO:9, CDR-L1 with SEQ ID NO:4, CDR- with SEQ ID NO:5 L2, CDR-L3 comprising SEQ ID NO:6, a light chain variable region comprising SEQ ID NO:8, and one or more of a light chain comprising the amino acid sequence set forth in SEQ ID NO:10. In certain of these embodiments, the FGFR3 antagonistic antibody is bofatamab. In other embodiments, the antagonistic FGFR3 antibody is selected from the group consisting of PRO-001 and IMC-D11. In certain embodiments, the FGFR3 inhibitor is a small molecule pan-FGFR inhibitor, and in certain of these embodiments, the pan-FGFR inhibitor is infigatinib, AZD4547, LY2874455, pemigatinib, BGJ398 , rogaratinib, PRN1371, Debio 1347, ARQ 087, and JNJ-42756493. In certain embodiments, the PD1 inhibitor binds PD1. In other embodiments, the PD1 inhibitor binds a ligand of PD1, eg, PDL1 or PDL2. In certain embodiments, the PD1 inhibitor is an antagonistic PD1 antibody, and in certain of these embodiments, the antagonistic PD1 antibody is nivolumab, pembrolizumab, CT-011, MEDI-0680 , and RMP1-14. In other embodiments, the PD1 inhibitor is an antagonistic PD1 ligand antibody, and in certain of these embodiments, the antagonistic PD1 ligand antibody is MEDI-4736, RG7446, BMS-936559, MSB0010718C, and MPDL3280A.

In certain embodiments, a method for treating a wild-type FGFR3-expressing cancer in a subject in need thereof, said method comprising administering to said subject a therapeutically effective amount of an FGFR3 inhibitor, Provided herein are methods comprising administering in combination with a therapeutically effective amount of a checkpoint inhibitor. In certain embodiments, the FGFR3 inhibitor binds FGFR3. In other embodiments, the FGFR3 inhibitor binds a ligand of FGFR3, eg, FGF1, FGF2, or FGF9. In certain embodiments, the FGFR3 inhibitor is an antagonistic FGFR3 antibody, and in certain of these embodiments, the antagonistic FGFR3 antibody comprises a CDR-H1 comprising SEQ ID NO: 1, SEQ ID NO: 2, CDR-H3 with SEQ ID NO:3, heavy chain variable region with SEQ ID NO:7, heavy chain with SEQ ID NO:9, CDR-L1 with SEQ ID NO:4, CDR- with SEQ ID NO:5 L2, CDR-L3 comprising SEQ ID NO:6, a light chain variable region comprising SEQ ID NO:8, and one or more of a light chain comprising the amino acid sequence set forth in SEQ ID NO:10. In certain of these embodiments, the FGFR3 antagonistic antibody is bofatamab. In other embodiments, the antagonistic FGFR3 antibody is selected from the group consisting of PRO-001 and IMC-D11. In certain embodiments, the FGFR3 inhibitor is a small molecule pan-FGFR inhibitor, and in certain of these embodiments, the pan-FGFR inhibitor is infigatinib, AZD4547, LY2874455, pemigatinib, BGJ398 , rogaratinib, PRN1371, Debio 1347, ARQ 087, and JNJ-42756493. In certain embodiments, the checkpoint inhibitor is a PD1 inhibitor. In certain of these embodiments, the PD1 inhibitor binds PD1. In other embodiments, the PD1 inhibitor binds a ligand of PD1, eg, PDL1 or PDL2. In certain embodiments, the PD1 inhibitor is an antagonistic PD1 antibody, and in certain of these embodiments, the antagonistic PD1 antibody is nivolumab, pembrolizumab, CT-011, MEDI-0680 , and RMP1-14. In other embodiments, the PD1 inhibitor is an antagonistic PD1 ligand antibody, and in certain of these embodiments, the antagonistic PD1 ligand antibody is MEDI-4736, RG7446, BMS-936559, MSB0010718C, and MPDL3280A.

In certain embodiments, a method for treating a subject in need thereof having a cancer that expresses wild-type FGFR3, said method comprising: (a) said subject; (b) whether said subject has a genetic signature that correlates with said one or more cancer-associated fibroblasts; or p53 expression, (c) based on the determination of step (b), (i) if said subject does not have said gene signature or p53 expression, a therapeutically effective amount of FGFR3 administering an inhibitor in combination with a therapeutically effective amount of a checkpoint inhibitor; and (ii) if said subject expresses said gene signature or p53 expression, a therapeutically effective amount of FGFR3 inhibitor. Provided herein are methods comprising administering in combination with an effective amount of a checkpoint inhibitor and an additional anti-cancer agent. In certain embodiments, the FGFR3 inhibitor is an antagonistic FGFR3 antibody. In certain embodiments, the antagonistic FGFR3 antibody has a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:1, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO:2, an amino acid sequence set forth in SEQ ID NO:3 and a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO:7. In certain embodiments, the antagonistic FGFR3 antibody has a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:4, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:5, an amino acid sequence set forth in SEQ ID NO:6 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO:8. In certain embodiments, the FGFR3 inhibitor is bofatamab. In certain embodiments, the checkpoint inhibitor is a PD1 inhibitor. In certain embodiments, the PD1 inhibitor is an antagonist PD-L1 antibody selected from the group consisting of MEDI-4736, RG7446, BMS-936559, MSB0010718C, and MPDL3280A. In certain embodiments, the PD1 inhibitor is pembrolizumab. In certain embodiments, the cancer is luminal bladder cancer. In certain embodiments, the gene signature comprises at least the following genes: FGFR3, TP63, IRS1, SEMA4B, PTPN13, and TMPRSS4. In certain embodiments, the gene signature comprises at least the following genes: KRT5, KRT6A, KRT6B, KRT14, UPK3A, UPK3B, FOXA1, and PPARG. In certain embodiments, the gene signature comprises at least the following genes: ACTC1, ACTG2, NCC1, DES, FLNC, MFAP4, MYH11, and PCP4.

In certain embodiments, provided herein are kits comprising FGFR3 inhibitors and checkpoint inhibitors (eg, PD1 inhibitors) for use in treating bladder cancer. In certain of these embodiments, the kit further comprises instructions for use.

In certain embodiments, provided herein is the use of FGFR3 inhibitors and checkpoint inhibitors (eg, PD1 inhibitors) for use in the formulation of medicaments for the treatment of bladder cancer. In certain of these embodiments, the FGFR3 inhibitor and PD1 inhibitor are formulated into one medicament. In other embodiments, the FGFR3 inhibitor and PD1 inhibitor are formulated into separate medicaments that are administered in combination with each other.

FIG. 1 shows the results of whole transcriptome RNAseq performed on 22 matched biopsies of tumors depicting two prognostic clusters pre- and post-bofatamab administration. Cluster 1 was mostly luminal in type and contained the majority of responders. Cluster 2 was more basal in nature and demonstrated clinical benefit consistent with expected response rates.

Figure 2 shows the results of whole transcriptome RNAseq performed on 22 matched biopsies of tumors. The genetic signatures used to identify basal and luminal subtypes were used to assign 22 paired tumors to molecular subtypes.

FIG. 3 shows alterations of inflammatory and immune pathways in responders after bofatamab treatment.

FIG. 4 is a schematic representation of the Phase 1b/Phase 2 study design investigating the effects of bofatamab plus pembrolizumab in lineage 2 mUC.

FIG. 5 shows RECIST 1.1 evaluable patient response in the WT vs. Mut/Fus based phase 2 trial.

FIG. 6 shows immune changes in responders.

FIG. 7 is a spider plot showing the change in summary lesion diameter (SoD) from baseline after bofatamab (B-701) treatment. This plot shows that bofatamab appears to induce an initial apparent increase in tumor size followed by a sharp reduction in tumor volume.

FIG. 8 illustrates new definitions of bladder cancer molecular subtypes.

Figure 9 shows RECIST 1.1 evaluable patient response in Phase 2 study based on molecular subgroups.

FIG. 10 shows the association between molecular subtypes and response to combination therapy.

Figure 11 shows the duration of treatment in the Phase 2 trial by molecular subgroup.

Figure 12 shows the results of combined bofatamab and pembrolizumab treatment in male mUC patients with luminal subtype.

FIG. 13 shows progression-free survival and overall survival of WT FGFR3 patients versus Mut/Fus patients.

Figure 14 shows survival after combined treatment with bofatamab and pembrolizumab. Patients with p53-like tumors (n=5) showed poor survival.

DETAILED DESCRIPTION The following description of the invention is intended only to illustrate various embodiments of the invention. Therefore, the specific modifications discussed should not be construed as limitations on the scope of the invention. It will be apparent to those skilled in the art that various equivalents, modifications and alterations can be made without departing from the scope of the invention, and such equivalent embodiments are intended to be included herein. is understood.

There are four single transmembrane tyrosine kinase fibroblast growth factor receptors (FGFR1-4) in humans (Brooks 2012). FGFR is overexpressed in many cancer types, often due to mutations that confer constitutive activation, making it an attractive target for therapeutic intervention. For example, the FGFR2b antibody FPA144 (FivePrime) is currently in development for the treatment of solid tumors, especially gastric cancer. Other FGFR2 monoclonal antibodies in early development for cancer treatment include GP369 (Aveo) and HuGAL-FR21 (Galaxy) (Zhao 2010; Bai 2010). Humanized anti-FGFR4 has also been reported to inhibit tumor growth (Bumbaca 2011).

FGFR3 has both oncogenic and tumor suppressor properties. FGFR3 is frequently mutated in certain cancers, but also in some normal tissues, where it can limit cell proliferation and promote cell differentiation (Lafitte 2013). The human FGFR3 antagonistic monoclonal antibody bofatamab (sometimes also referred to as B-701 or BM2) was the first FGFR3-targeting antibody to enter clinical development. The variable portion of bofatamab was originally identified through phage display and then recombined with a human IgG1 backbone. Bofatamab ^is effective against both wild- ^type and mutant FGFR3, the most common mutations found in bladder cancer and ^{achondroplasia (} particularly FGFR3-IIIb ^G372C ) and gene fusions containing FGFR3 TACC3 and FGFR3 BAIAP2L1), but binds to other FGFRs without cross-reactivity. Bofatamab was previously evaluated for safety in patients with t(4:14) translocation multiple myeloma (Clinical Trial NCT01122875). Other FGFR3 inhibitor antibodies that are in or have been in clinical or preclinical development include: PRO-001 (Prochon), IMC-D11 (ImClone), and ADC LY3076226 (Eli Lilly). Additional FGFR3 antibodies for use in treating cancer and other diseases are disclosed, for example, in US Pat. Nos. 8,187,601 (Aveo) and 7,498,416 (Fibron). there is

Programmed cell death protein 1 (PD1) is an immune checkpoint receptor from the CD28 superfamily that limits T cell effector function in tissues following activation by either of its two ligands PDL1 or PDL2 ( Pardoll 2012). PD1 downregulates the immune system by promoting apoptosis in antigen-specific T cells while simultaneously reducing apoptosis in regulatory (ie, suppressor) T cells. Certain tumor cells block anti-tumor immune responses in the tumor microenvironment by upregulating PD1 ligands. Blocking the PD1 pathway has been shown to activate the immune system to attack tumors and induce sustained tumor regression in various tumor types. Accordingly, several PD1 antagonist antibodies are currently approved or in various stages of clinical development. For example, the fully human IgG4 monoclonal PD1 antibody, nivolumab (also known as ^Opdivo® , Bristol-Myers Squibb and Ono Pharmaceutical; ONO-4538, BMS-936558, MDX-1106) no longer responds to other drugs. Approved for the treatment of non-selective or metastatic melanoma in patients. Nivolumab is also being evaluated for the treatment of non-small cell lung cancer (NSCLC) in combination with various chemotherapy regimens. Pembrolizumab ( ^Keytruda® , Merck; also known as MK-3475), a humanized IgG4 PD1 antibody, is approved for the treatment of several types of cancer, including UCC, NSCLC, and melanoma. ing. Other PD1 antibodies in development include CT-011 (Curetech) and MEDI-0680/AMP-514 (AstraZeneca). is mentioned.

Various PD1 ligand (PDL) antibodies are also in development for cancer treatment. For example, the monoclonal IgG1k PDL1 antibody MEDI-4736 (AstraZeneca) is currently in development for the treatment of NSCLC alone or in combination with the monoclonal CTLA4 antibody tremelimumab (AstraZeneca) or MEDI-0680, and the monoclonal IgG1k PDL1 antibody RG7446 (Roche ) is in development for use in treating various cancers alone or in combination with ^Avastin® and ^Zelboraf® , a fully human monoclonal IgG4 antibody BMS-936559/MDX-1105 (BMS) is currently in development for the treatment of NSCLC and other cancer types, the fully human IgG1 PDL1 antibody MSB0010718C (Merck Serono) is in development for the treatment of various cancer types, and the Fc-engineered monoclonal IgG1 Antibody MPDL3280A (Genentech) is currently in development for the treatment of NSCLC.

As shown in the experimental results provided herein, the FGFR3 inhibitor bofatamab has been shown to induce activation of genes associated with key signaling molecules in TH1 responses and immune function. These results suggest that bofatamab activates genes associated with immune cell trafficking pathways. The results also suggest that bofatamab sensitizes patients with mUC in luminal biology to combined treatment with bofatamab and checkpoint inhibitors (eg, PD1 inhibitors). The results presented herein demonstrate that a genetic signature associated with cancer-associated fibroblasts appears to be associated with resistance to combined bofatamab and checkpoint inhibitors, and that patients with p53-like tumors also It is further shown that the combination treatment was tolerated. It is a candidate for combination therapy in which patients exhibiting a cancer-associated fibroblast gene signature or with p53-like tumors further include a third agent, such as an anti-fibrotic agent. Suggest to get

In certain embodiments, provided herein are methods of treating cancer in a subject in need thereof comprising administering an FGFR3 inhibitor and a PD1 inhibitor. 1. A method of increasing the efficacy of a PD1 inhibitor to treat cancer in a subject in need thereof, said method comprising administering an FGFR3 inhibitor and treating cancer in a subject in need thereof. Also provided herein is a method of increasing the efficacy of an FGFR3 inhibitor for treating of, the method comprising administering a PD1 inhibitor. An increase in the efficacy of a PD1 or FGFR3 inhibitor is an increase in the therapeutic effect of either inhibitor, the required dosage, frequency of administration, or interval between administrations of either inhibitor to obtain a specified level of therapeutic effect. or some combination thereof.

In certain embodiments, the methods provided herein are used to treat solid tumors, ie, cancers that form discrete tumor masses. In certain of these embodiments, the cancer being treated is bladder cancer, including, eg, metastatic bladder cancer (mUC) or upper urothelial cancer.

The terms "treat," "treating," and "treatment," as used herein with respect to solid tumors, partially or completely inhibit tumor growth, tumor It may refer to size reduction, complete or partial tumor eradication, reduction or prevention of malignant growth, partial or complete eradication of cancer cells, or some combination thereof. The terms "patient" and "subject" are used interchangeably herein.

A "subject in need thereof," as used herein, is a person diagnosed with cancer, suspected of having cancer, and/or associated with cancer. Reference is made to mammalian subjects, preferably humans, who exhibit more symptoms than this. In certain embodiments, the subject may have one or more previously received therapeutic interventions, such as chemotherapy, for the treatment of cancer.

"FGFR3 inhibitor" as used herein refers to any molecule that either partially or completely inhibits the activity of FGFR3. FGFR3 inhibitors may specifically inhibit FGFR3, or may inhibit the activity of other proteins in addition to FGFR3. For example, FGFR3 inhibitors can also inhibit the activity of other FGFRs. In certain embodiments, FGFR3 inhibitors may exhibit indirect immunomodulatory activity. For example, FGFR3 inhibitors can have indirect effects on immune cell trafficking pathways, eg, by altering the expression and/or activity of TNFα or IFNγ.

An "antagonist antibody", as used herein, refers to a receptor and its cognate ligand by physical binding to the receptor or its cognate ligand at a binding site and/or an allosteric site on any molecule. It refers to an antibody that reduces, prevents, or otherwise inhibits interaction between its cognate ligand. In some embodiments, the antagonist antibody interacts at a unique binding site that is not otherwise involved in biological modulation of receptor activity by the cognate ligand. Thus, an antagonist antibody has affinity for the receptor or the cognate ligand, but is less effective in promoting a biological response once bound compared to the cognate ligand binding to the receptor. do not. Binding of an antagonist antibody thereby prevents interaction between the receptor and the cognate ligand, otherwise inhibiting agonist function.

In certain embodiments of the methods, compositions, and kits provided herein, the FGFR3 inhibitor inhibits FGFR3 activity by binding to FGFR3. Examples of such FGFR3 inhibitors include, for example, antagonistic FGFR3 antibodies or fusion proteins thereof, inactive forms of said FGFR3 ligands (e.g. truncated or otherwise mutated forms of FGFR3 ligands) or fusion proteins thereof. , small molecules, siRNAs, and aptamers. In certain of these embodiments, the FGFR3 inhibitor specifically binds FGFR3. This means that the inhibitors show little or no binding to other FGFRs. In other embodiments, the FGFR3 inhibitor binds one or more FGFRs in addition to FGFR3.

In certain preferred embodiments of the methods, compositions, and kits provided herein, the FGFR3 inhibitor is an FGFR3 antagonist antibody, and in certain of these embodiments, the FGFR3 antagonist The antibody specifically binds FGFR3. The term "antibody" as used herein refers to an immunoglobulin molecule or immunologically active portion thereof that binds to a specific antigen (eg, FGFR3 or PD1). In those embodiments where the antibody for use in the methods, compositions, and kits of the invention is a full-length immunoglobulin molecule, the antibody is two heavy chains and two light chains, each heavy chain and light chains containing three complementarity determining regions (CDRs). In those embodiments where the antibody is an immunologically active portion of an immunoglobulin molecule, the antibody is, for example, Fab, Fab', Fv, Fab', F(ab')2, disulfide-linked Fv, scFv , single domain antibodies (dAbs), or diabodies. Antibodies for use in the methods, compositions, and kits of the invention include natural antibodies, synthetic antibodies, monoclonal antibodies, polyclonal antibodies, chimeric antibodies, humanized antibodies, multispecific antibodies, bispecific antibodies ( bispecific antibodies, dual-specific antibodies, anti-idiotypic antibodies, or fragments thereof that retain the ability to bind a specific antigen (eg, FGFR3 or PD1). Exemplary antibodies include IgA, IgD, IgG1, IgG2, IgG3, IgM, and the like. In certain preferred embodiments of the methods, compositions, and kits provided herein, the FGFR3 antibody is an IgG2 antibody.

In certain embodiments, the FGFR3 antagonist antibodies for use in the methods, compositions, and kits of the invention comprise one or more complementarity determining regions (CDRs) having the sequences set forth in SEQ ID NOS: 1-3. ). In certain of these embodiments, the FGFR3 antagonist antibody comprises all three of these CDR sequences, and in certain of these embodiments, the FGFR3 antagonist antibody comprises , a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO:4. In certain embodiments, the FGFR3 antagonist antibody comprises a light chain variable region comprising one or more CDRs having sequences set forth in SEQ ID NOS:5-7. In certain of these embodiments, the FGFR3 antagonist antibody comprises all three of these CDR sequences, and in certain of these embodiments, the FGFR3 antagonist antibody comprises , a light chain variable region comprising the amino acid sequence shown in SEQ ID NO:8. In certain embodiments, the FGFR3 antagonist antibody comprises all six CDR sequences set forth in SEQ ID NOs: 1-3 and 5-7, and in certain of these embodiments, the FGFR3 antagonist antibody The antibody comprises a heavy chain variable region of SEQ ID NO:4 and a light chain variable region of SEQ ID NO:8. In certain embodiments, the antibody is bofatamab comprising the heavy chain of SEQ ID NO:9 and the light chain of SEQ ID NO:10. The heavy chain of SEQ ID NO:9, in addition to the variable region shown in SEQ ID NO:7, constitutes human IgG1. Similarly, the light chain of SEQ ID NO: 10 comprises the variable region shown in SEQ ID NO: 8 and human Igκ chain C (UniProt P01834).

SEQ ID NO: 1 (H1-CDR): GFTFTSTGIS.

SEQ ID NO: 2 (H2-CDR): GRIYPTSGSTNYADSVKG.

SEQ ID NO: 3 (H3-CDR): ARTYGIYDLYVDYTEYVMDY.

SEQ ID NO: 4 (L1-CDR): RASQDVDTSLA.

SEQ ID NO: 5 (L2-CDR): SASFLYS.

SEQ ID NO: 6 (L3-CDR): QQSTGHPQT.

SEQ ID NO:7:
EVQLVESGGGGLVQPGGSLRLSCAASGFTFFTSTGISWVRQAPGKGLEWVGRIYPTSGSTNYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARTYGIYDLYVDYTEYVMDYWGQGTLV.

SEQ ID NO:8:
DIQMTQSPSSLSASVGDRVTITCRASQDVDTSLAWYKQKPGKAPKLLIYSASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSTGHPQTFGQGTKVEIKR.

SEQ ID NO:9:
ＥＶＱＬＶＥＳＧＧＧＬＶＱＰＧＧＳＬＲＬＳＣＡＡＳＧＦＴＦＴＳＴＧＩＳＷＶＲＱＡＰＧＫＧＬＥＷＶＧＲＩＹＰＴＳＧＳＴＮＹＡＤＳＶＫＧＲＦＴＩＳＡＤＴＳＫＮＴＡＹＬＱＭＮＳＬＲＡＥＤＴＡＶＹＹＣＡＲＴＹＧＩＹＤＬＹＶＤＹＴＥＹＶＭＤＹＷＧＱＧＴＬＶＴＶＳＳＡＳＴＫＧＰＳＶＦＰＬＡＰＳＳＫＳＴＳＧＧＴＡＡＬＧＣＬＶＫＤＹＦＰＥＰＶＴＶＳＷＮＳＧＡＬＴＳＧＶＨＴＦＰＡＶＬＱＳＳＧＬＹＳＬＳＳＶＶＴＶＰＳＳＳＬＧＴＱＴＹＩＣＮＶＮＨＫＰＳＮＴＫＶＤＫＫＶＥＰＫＳＣＤＫＴＨＴＣＰＰＣＰＡＰＥＬＬＧＧＰＳＶＦＬＦＰＰＫＰＫＤＴＬＭＩＳＲＴＰＥＶＴＣＶＶＶＤＶＳＨＥＤＰＥＶＫＦＮＷＹＶＤＧＶＥＶＨＮＡＫＴＫＰＲＥＥＱＹＮＳＴＹＲＶＶＳＶＬＴＶＬＨＱＤＷＬＮＧＫＥＹＫＣＫＶＳＮＫＡＬＰＡＰＩＥＫＴＩＳＫＡＫＧＱＰＲＥＰＱＶＹＴＬＰＰＳＲＥＥＭＴＫＮＱＶＳＬＴＣＬＶＫＧＦＹＰＳＤＩＡＶＥＷＥＳＮＧＱＰＥＮＮＹＫＴＴＰＰＶＬＤＳＤＧＳＦＦＬＹＳＫＬＴＶＤＫＳＲＷＱＱＧＮＶＦＳＣＳＶＭＨＥＡＬＨＮＨＹＴＱＫＳＬＳＬＳＰＧＫ．

SEQ ID NO: 10:
ＤＩＱＭＴＱＳＰＳＳＬＳＡＳＶＧＤＲＶＴＩＴＣＲＡＳＱＤＶＤＴＳＬＡＷＹＫＱＫＰＧＫＡＰＫＬＬＩＹＳＡＳＦＬＹＳＧＶＰＳＲＦＳＧＳＧＳＧＴＤＦＴＬＴＩＳＳＬＱＰＥＤＦＡＴＹＹＣＱＱＳＴＧＨＰＱＴＦＧＱＧＴＫＶＥＩＫＲＴＶＡＡＰＳＶＦＩＦＰＰＳＤＥＱＬＫＳＧＴＡＳＶＶＣＬＬＮＮＦＹＰＲＥＡＫＶＱＷＫＶＤＮＡＬＱＳＧＮＳＱＥＳＶＴＥＱＤＳＫＤＳＴＹＳＬＳＳＴＬＴＬＳＫＡＤＹＥＫＨＫＶＹＡＣＥＶＴＨＱＧＬＳＳＰＶＴＫＳＦＮＲＧＥＣ．

In other embodiments, the FGFR3 antagonist antibody for use in the methods, compositions, and kits of the invention is PRO-001, IMC-D11, or US Pat. No. 8,187,601 (Aveo) or US Pat. FGFR3 antagonistic antibodies as disclosed in US Pat. No. 7,498,416 (Fibron). In certain embodiments, FGFR3 antagonist antibodies for use in the methods, compositions, and kits of the invention can be lyophilized.

In certain embodiments of the methods, compositions, and kits provided herein, the FGFR3 inhibitor inhibits FGFR3 activity by binding to a ligand of FGFR3, eg, FGF1, FGF2, or FGF9. Examples of such FGFR3 inhibitors include, for example, antibodies that specifically bind FGFR3 ligands or fusion proteins thereof, soluble forms of FGFR3 comprising all or part of the FGFR3 extracellular domain or fusion proteins thereof, downstream signaling truncated forms of FGFR3 or fusion proteins thereof, small molecules, siRNAs, and aptamers lacking all or part of the intracellular domain required for .

In certain embodiments of the methods, compositions, and kits provided herein, the FGFR3 inhibitor is a pan-FGFR inhibitor, which means that the pan-FGFR inhibitor comprises, in addition to FGFR3, one or more It means binding to many FGFRs and inhibiting their activity. In certain of these embodiments, the FGFR3 inhibitor is Infigratinib (BGJ398, Novartis), AZD4547 (AstraZeneca), LY2874455 (Eli Lilly), Debio 1347 (Debiopharm), ARQ 087 (ArQule), A small molecule pan-FGFR inhibitor selected from the group consisting of JNJ-42756493 (Janssen), and PRN1371 (Principia).

In certain embodiments of the methods, compositions, and kits provided herein, the FGFR3 inhibitor inhibits FGFR3 activity by blocking downstream tyrosine kinase activity. For example, non-selective tyrosine kinase inhibitors such as dovitinib, lucitinib, ponatinib, nintedanib, ponatinib, or ENMD-2076 can be utilized as FGFR3 inhibitors.

"PD1 inhibitor" as used herein refers to any molecule that inhibits, either partially or completely, the activity of PD1. A PD1 inhibitor may specifically inhibit PD1, or may inhibit the activity of other proteins in addition to PD1. For example, PD1 inhibitors may also inhibit the activity of other immune checkpoint molecules.

In certain embodiments of the methods, compositions, and kits provided herein, the PD1 inhibitor inhibits PD1 activity by binding to PD1. Examples of such PD1 inhibitors include, for example, antagonistic PD1 antibodies or fusion proteins thereof, inactive forms of PD1 ligand (e.g. truncated or otherwise mutated forms of PDL1 or PDL2) or fusion proteins thereof (e.g. , AMP-224 (GlaxoSmithKline, Amplimmune)), small molecules, siRNAs, and aptamers.

In certain embodiments of the methods, compositions, and kits provided herein, the PD1 inhibitor is a PD1 antibody, and in certain of these embodiments, the PD1 antagonist antibody is , specifically binds PD1. In certain embodiments, the PD1 antagonistic antibody is selected from the group consisting of nivolumab, pembrolizumab, CT-011, MEDI-0680, and RMP1-14.

In certain embodiments of the methods, compositions, and kits provided herein, the PD1 inhibitor reduces PD1 activity by binding to one or more ligands of PD1 (i.e., PDL1 or PDL2). impede Examples of such PD1 inhibitors include, for example, PD1 ligand antibodies or fusion proteins thereof, soluble forms of PD1 comprising all or part of the PD1 extracellular domain or fusion proteins thereof, cells required for downstream signaling. Included are truncated forms of PD1 lacking all or part of the endodomain or fusion proteins thereof, small molecules, siRNAs, and aptamers.

In certain embodiments of the methods, compositions, and kits provided herein, the PD1 inhibitor is a PD1 ligand antibody, and in certain of these embodiments, the PD1 ligand antibody specifically binds the PD1 ligand. In certain embodiments, said PD1 ligand antibody is selected from the group consisting of MEDI-4736, RG7446, BMS-936559, MSB0010718C, and MPDL3280A.

In certain embodiments of the methods provided herein, the FGFR3 inhibitor and PD1 inhibitor are administered together as part of the same composition. In other embodiments, the FGFR3 inhibitor and PD1 inhibitor are administered separately, ie, as separate compositions. In these embodiments, the inhibitors may be administered simultaneously or sequentially, and may be administered via the same route of administration or different routes of administration. In those embodiments in which the inhibitors are administered sequentially, they are They may be administered at the same or different intervals. For example, one inhibitor may be administered more frequently than the other, or may be administered over a longer course. In certain of these embodiments, one inhibitor precedes the first administration of the second inhibitor. It may be administered one or more times. When administration of the second inhibitor is initiated, administration of the first inhibitor may be discontinued or continued throughout all or part of the course of administration of the second inhibitor. In certain embodiments, wherein the FGFR3 inhibitor is an FGFR3 antagonist antibody, the antibody is administered twice or more per day, twice per week or more, once per week for two weeks. It can be administered once a day (ie, every other week), once every three weeks, or once a month. In certain embodiments, the antibody is administered once a week, once every two weeks, or once every three weeks. In certain embodiments, wherein the PD1 inhibitor is a PD1 antagonist antibody, the antibody is administered two or more times per day, two or more times per week, two or more times per week, two It can be administered once a week (ie, every other week), once every three weeks, or once a month. In certain embodiments, the PD1 inhibitor is administered once every two weeks. In certain embodiments, the FGFR3 inhibitor and/or the PD1 inhibitor may be administered over a predetermined course. For example, the FGFR3 and/or PD1 inhibitor can be administered over the course of 1 day, 2 days, 1 week, 2 weeks, 4 weeks, or 8 weeks. In other embodiments, the FGFR3 and/or PD1 inhibitor can be administered indefinitely or until a specific therapeutic benchmark is achieved. For example, the FGFR3 and/or PD1 inhibitor is administered until tumor growth is arrested or ameliorated, until one or more tumors are eliminated, or until the number of cancer cells is reduced to a certain level. can be

A "therapeutically effective amount" of a composition, as used herein, is that amount of the composition that produces the desired therapeutic effect in a subject, such as one being treated for cancer. . In certain embodiments, the therapeutically effective amount is the amount of the composition that produces maximal therapeutic effect. In other embodiments, the therapeutically effective amount produces a therapeutic effect that is less than the maximal therapeutic effect. For example, a therapeutically effective amount can be an amount that produces therapeutic effect while avoiding one or more side effects associated with doses that produce maximal therapeutic effect. A therapeutically effective amount for a particular composition depends on a variety of factors, including the characteristics of the therapeutic composition (e.g., activity, pharmacokinetics, pharmacodynamics, and bioavailability), the physiological state of the subject (e.g., age, body weight, sex, disease type and stage, medical history, general physical condition, responsiveness to prescribed doses, and other current medications), any pharmaceutically acceptable carrier in the composition. nature, and route of administration, including but not limited to). Those of ordinary skill in the clinical and pharmacological arts will determine a therapeutically effective amount through routine experimentation, i.e., by monitoring the subject's response to administration of the composition and adjusting the dosage accordingly. can decide.さらなるガイダンスについては、例えば、Ｒｅｍｉｎｇｔｏｎ： Ｔｈｅ Ｓｃｉｅｎｃｅ ａｎｄ Ｐｒａｃｔｉｃｅ ｏｆ Ｐｈａｒｍａｃｙ， 第２２版， Ｐｈａｒｍａｃｅｕｔｉｃａｌ Ｐｒｅｓｓ， Ｌｏｎｄｏｎ， ２０１２、およびＧｏｏｄｍａｎ ＆ Ｇｉｌｍａｎ'ｓ Ｔｈｅ Ｐｈａｒｍａｃｏｌｏｇｉｃａｌ Ｂａｓｉｓ ｏｆ Ｔｈｅｒａｐｅｕｔｉｃｓ， 第１２版， ＭｃＧｒａｗ－Ｈｉｌｌ， Ｎｅｗ Ｙｏｒｋ， NY, 2011, the entire disclosures of which are incorporated herein by reference.

In certain embodiments of the methods provided herein, a therapeutically effective amount of a FGFR3 inhibitor or PD1 inhibitor provides a therapeutic response ( For example, it can be a dosage that can produce a reduction or elimination of tumor growth. In certain of these embodiments, the therapeutically effective amount can be a dose previously determined to be optimal or near optimal for cancer treatment. For example, when the FGFR3 inhibitor is bofatamab, the antibody may be administered at a dose of about 10-50 mg/kg every 2-4 weeks; may be administered at a dose of about 20-40 mg/kg every 2-4 weeks, or at a dose of about 30 mg/kg every 3 weeks. In other embodiments, the therapeutically effective amount of the FGFR3 inhibitor or PD1 inhibitor may be lower than the dose at which the molecule would normally be administered for use as monotherapy (i.e., a suboptimal dose). . In certain of these embodiments, administration of suboptimal doses of FGFR3 or PD1 inhibitors may result in reduced side effects relative to standard doses when administered alone. For example, administration of suboptimal doses of FGFR3 or PD1 inhibitors may result in a reduced incidence or severity of pruritus, colitis, or pneumonia relative to administration of optimal doses of either inhibitor alone. In certain embodiments, one of the FGFR3 inhibitor and the PD1 inhibitor may be administered at a dosage determined to be optimal for cancer treatment when administered alone, while the other is administered alone. It is administered in dosages that are less than optimal for treatment when administered. In certain embodiments, the dosage of the FGFR3 inhibitor or PD1 inhibitor may vary over the course of the treatment regimen. For example, one or both of the FGFR3 inhibitor and PD1 inhibitor can be administered at a higher dosage at the beginning of treatment (e.g., a loading phase), followed by a lower dosage later in treatment. .

Compositions comprising FGFR3 inhibitors, PD1 inhibitors, or both FGFR3 inhibitors and PD1 inhibitors can be administered by any route of administration known in the art (parenteral, oral, aerosol, enteral, nasal, ophthalmic, parenteral, or oral). It can be delivered to the subject by skins (eg, topical creams or ointments, patches, including but not limited to). "Parenteral" means routes of administration commonly associated with injection (intravenous, intraperitoneal, subcutaneous, suborbital, infusion, intraarterial, intracapsular, intracardiac, intradermal, intramuscular, intrapulmonary, intrathecal intrasternal, intrasternal, intratracheal, intrauterine, subarachnoid, subcapsular, transmucosal, or transtracheal). In certain embodiments, wherein the FGFR3 inhibitor is an FGFR3 antagonist antibody (eg, including bofatamab), the FGFR3 inhibitor is administered intravenously. In certain embodiments, wherein the PD1 inhibitor is a PD1 antagonist antibody, the PD1 inhibitor is administered intraperitoneally.

In certain embodiments, compositions comprising FGFR3 inhibitors, PD1 inhibitors, or both FGFR3 and PD1 inhibitors can be formed into oral dosage units such as tablets, pills, or capsules. In certain embodiments, an FGFR3 inhibitor, PD1 inhibitor, or FGFR3 and PD1 inhibitor composition can be administered via a time-release delivery vehicle, such as a time-release capsule. A "time release vehicle" as used herein refers to any delivery vehicle that releases an active agent over a period of time rather than immediately upon administration. In other embodiments, the FGFR3 inhibitor, PD1 inhibitor, or FGFR3 and PD1 inhibitor composition can be administered via an immediate release delivery vehicle.

In certain embodiments of the methods provided herein, the subject receiving the FGFR3 inhibitor and the PD1 inhibitor is given additional therapy ( chemotherapy or immunotherapy), or a third agent (eg, an antifibrotic agent). In those embodiments in which the subject receives additional therapy during treatment with an FGFR3 and PD1 inhibitor, the additional therapy may be administered concurrently or sequentially with the FGFR3 inhibitor and/or PD1 inhibitor.

In certain embodiments, provided herein are compositions comprising a therapeutically effective amount of a FGFR3 inhibitor and a therapeutically effective amount of a PD1 inhibitor. In certain embodiments, these compositions further comprise one or more pharmaceutically acceptable carriers or for administration with one or more pharmaceutically acceptable carriers. It is formulated into Also provided herein are kits containing an FGFR3 inhibitor and a PD1 inhibitor for use in performing the methods disclosed herein, eg, to treat cancer.

In certain embodiments of the compositions and kits provided herein, the FGFR3 inhibitor or PD1 inhibitor can produce a therapeutic response (e.g., reduce or eliminate tumor growth) when administered alone In dosage, it can be present in the composition or kit described above. In certain of these embodiments, the FGFR3 or PD1 inhibitor may be present at a dosage previously determined to be optimal or near optimal for cancer treatment. For example, if the FGFR3 inhibitor is bofatamab, the composition or kit can be formulated to deliver a dose of about 10-50 mg/kg of bofatamab to the subject; In certain of them, the composition or kit can be formulated to deliver a dose of bofatamab of about 20-40 mg/kg or about 30 mg/kg to the subject. In other embodiments, the FGFR3 or PD1 inhibitor may be present at a lower dosage than it would normally be present in a composition or kit for treating cancer (ie, a suboptimal dosage).

"Pharmaceutically acceptable carrier" as used herein means a carrier from one tissue, organ, or part of the body to another tissue, organ, or part of the body; Refers to a pharmaceutically acceptable substance, composition, or vehicle involved in carrying or transporting a compound or molecule of interest. Pharmaceutically acceptable carriers include various components (liquid or solid fillers, diluents, excipients, solvents, buffers, encapsulating agents, surfactants, stabilizers, binders, or pigments, or some combination thereof, including but not limited to. Each component of the carrier must be compatible with the other ingredients of the composition and suitable for contact with any tissue, organ, or part of the body it may encounter. "pharmaceutically acceptable" in that each component of the carrier exhibits toxicity, irritation, allergic response, immunogenicity, or other adverse effects that unduly outweigh its therapeutic benefits. should not be at risk of other complications of

Examples of pharmaceutically acceptable carriers that can be used with the compositions provided herein include, but are not limited to: (1) sugars (e.g., lactose, glucose, sucrose, or mannitol); (2) starches (e.g. corn starch and potato starch); (3) cellulose and its derivatives (e.g. sodium carboxymethylcellulose, ethylcellulose and cellulose acetate); (4) powdered tragacanth; (8) excipients such as cocoa butter and suppository waxes; (9) oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean. (10) glycols (e.g. propylene glycol); (11) polyols (e.g. glycerin, sorbitol, mannitol and polyethylene glycol); (12) esters (e.g. ethyl oleate and ethyl aurate). (13) disintegrants such as agar or calcium carbonate; (14) buffers or pH modifiers such as magnesium hydroxide, aluminum hydroxide, sodium chloride, sodium lactate, calcium chloride, and phosphate buffers; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) alcohols such as ethyl alcohol and propane alcohol; (21) lubricants (e.g. talc, calcium stearate, magnesium stearate, solid polyethylene glycol, or sodium lauryl sulfate); (22) colorants or pigments; (23) lubricants (e.g. colloidal silicon dioxide, talc, and starch or tribasic calcium phosphate); (24) other non-toxic compatible substances used in pharmaceutical compositions (eg, acetone); and (25) combinations thereof.

Compositions comprising FGFR3 inhibitors, PD1 inhibitors, or combinations of FGFR3 inhibitors and PD1 inhibitors may be prepared in suitable dosage forms, such as solutions or suspensions in aqueous or non-aqueous liquids, oil-in-water or water-in-oil liquids. emulsions, capsules, cachets, pills, tablets, lozenges, powders, granules, elixirs or syrups, or pastilles). In certain embodiments, the composition can be formulated as a time-release delivery vehicle, such as a time-release capsule. A "time-release vehicle," as used herein, refers to any delivery vehicle that releases an active agent over a period of time rather than immediately upon administration. In other embodiments, the composition can be formulated via an immediate release delivery vehicle.

In certain embodiments, provided herein are kits for performing the methods disclosed herein. In certain embodiments, kits provided herein comprise an FGFR3 inhibitor and a PD1 inhibitor. In certain embodiments, the FGFR3 inhibitor and PD1 inhibitor may be present in a kit in a single composition. In other embodiments, the FGFR3 inhibitor and PD1 inhibitor may be present in separate compositions. The kit may contain additional therapeutic or non-therapeutic compositions. In certain embodiments, the kit includes instructions in a tangible medium.

In certain embodiments, provided herein are FGFR3 inhibitors and PD1 inhibitors for use in treating cancer. Provided herein are FGFR3 inhibitors for use in treating cancer in combination with PD1 inhibitors, and PD1 inhibitors for use in treating cancer in combination with FGFR3 inhibitors. In certain of these embodiments, the cancer is urothelial carcinoma (eg, mUC). In certain of these embodiments, the cancer is luminal bladder cancer. For example, a subject in need of treatment for cancer with a particular genetic signature may be a candidate for treatment with FGFR3 inhibitors and PD1 inhibitors of the present disclosure. As another example, a subject in need of treatment with a cancer having another particular genetic signature may not be a candidate for treatment with the FGFR3 inhibitors and PD1 inhibitors of this disclosure, but rather You may have a treatment that includes three drugs. In some embodiments, the certain other specific gene signatures include gene signatures associated with cancer-associated fibroblasts and p53-like tumors. In certain embodiments, the gene signature is the FGFR3 gene signature and includes at least the following genes: FGFR3, TP63, IRS1, SEMA4B, PTPN13, and TMPRSS4. In certain embodiments, the gene signature is a p53-like gene signature and includes at least the following genes: KRT5, KRT6A, KRT6B, KRT14, UPK3A, UPK3B, FOXA1, and PPARG. In certain embodiments, the gene signature is a cancer-associated fibroblast (CAF) gene signature and includes at least the following genes: ACTC1, ACTG2, NCC1, DES, FLNC, MFAP4, MYH11, and PCP4.

In certain embodiments, a method for treating a subject in need thereof having a cancer that expresses wild-type FGFR3, said method comprising: (a) said subject; (b) the subject has a genetic signature that correlates with one or more cancer-associated fibroblasts or p53 (c) based on the determination of step (b), if (i) said subject does not have said gene signature or p53 expression, a therapeutically effective amount of an FGFR3 inhibitor; administering in combination with a therapeutically effective amount of a checkpoint inhibitor, and (ii) if said subject has a gene signature or p53 expression, a therapeutically effective amount of a FGFR3 inhibitor, and a therapeutically effective amount of a checkpoint inhibitor; Methods are provided herein that include administering in combination with a point inhibitor and an additional anticancer agent. In certain embodiments, the FGFR3 inhibitor is an antagonistic FGFR3 antibody. In certain embodiments, the antagonistic FGFR3 antibody has a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:1, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO:2, an amino acid sequence set forth in SEQ ID NO:3 and a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO:7. In certain embodiments, the antagonistic FGFR3 antibody has a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:4, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:5, an amino acid sequence set forth in SEQ ID NO:6 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO:8. In certain embodiments, the FGFR3 inhibitor is bofatamab. In certain embodiments, the checkpoint inhibitor is a PD1 inhibitor. In certain embodiments, the PD1 inhibitor is an antagonist PD-L1 antibody selected from the group consisting of MEDI-4736, RG7446, BMS-936559, MSB0010718C, and MPDL3280A. In certain embodiments, the PD1 inhibitor is pembrolizumab. In certain embodiments, the cancer is luminal bladder cancer.

In certain embodiments, provided herein is the use of FGFR3 inhibitors and PD1 inhibitors in the manufacture of a medicament for treating cancer. The use of a FGFR3 inhibitor in the manufacture of a medicament for treating cancer in combination with a PD1 inhibitor and the use of a PD1 inhibitor in the manufacture of a medicament for treating cancer in combination with a FGFR3 inhibitor are also provided herein. provided in

The term "about," as used herein, means within 10% of the stated value or range of values.

Those skilled in the art will recognize that the various embodiments described herein can be combined. For example, steps from various methods of treatment disclosed herein may be combined to achieve a satisfactory or improved level of treatment.

From the foregoing, it will be appreciated that although specific embodiments of the invention have been described herein for purposes of illustration, various modifications can be made without departing from the scope of the invention. Accordingly, the invention is not limited except as by the appended claims.

The following examples are provided to better illustrate the claimed invention and should not be construed as limiting the scope of the invention. To the extent specific materials are mentioned, the examples are for illustrative purposes only and are not intended to limit the invention. A person skilled in the art may develop equivalent means or reactants without exercising inventive capacity and without departing from the scope of the invention. It is understood that many variations can be made in the procedures described herein while still remaining within the scope of the invention. It is the intent of the inventors that such variations are included within the scope of the present invention.

Example 1: Gene expression profiles in wild-type and mutated FGFR3 metastatic urothelial carcinoma treated with combination therapy with bofatamab and pembrolizumab FGFR3 mutations among patients with metastatic urothelial carcinoma (mUC) (mutFGFR3) is present in 15-20% of bladder cancer patients and approximately 35% of upper urinary tract cancers. Studies suggest that these tumors may respond better to FGFR inhibition than to immunotherapy.

Patients who had failed previous treatments for mUC were treated with bofatamab. Patients were biopsied before and 14 days after treatment and treated with a loading dose of bofatamab (25 mg/kg) followed by cohorts of patients with and without mutFGFR3 or gene injection in combination with pembrolizumab. underwent therapy. Core biopsies (2×18 G or 3-4×20 G) were fixed in 10% neutral buffered formalin and embedded in paraffin. The biopsy cores were then isolated from paraffin blocks of these formalin-fixed paraffin-embedded (FFPE) samples.

DNA and RNA were isolated from samples for whole transcriptome RNAseq and DNA sequencing on the Ion Torrent platform. Whole-transcriptome RNAseq was performed using Ion Torrent's AmpliSeq RNA platform (Thermo Fisher, Inc.) and the Ion Proton sequencer (Thermo Fisher, Inc.) to match pre- and post-treatment biopsies Samples from 22 patients (17 WT, 5 Mut Fusion) were performed. RNA was then transcribed into cDNA using the ^{SuperScript®} VILO ^™ kit. cDNA was amplified using the Ion Ampliseq Transcriptome Human Gene Expression Core panel, followed by ligation of adapters and barcodes into amplicons and purification. Purified libraries were quantified using the Ion Library Quantification kit (Thermo Fisher, Inc.). The library was then diluted to 100 pM, pooled, amplified on Ion Sphere ^™ particles (ISP) using emulsion PCR, and enriched for IonChef (Thermo Fisher, Inc). Template-positive ISPs were then loaded onto an Ion PI chip and run on the Proton instrument. Targeted DNA sequencing of cancer-related genes was performed using the AmpliSeq Comprehensive Cancer panel (oncogenes including 409 tumor suppressor genes and FGFR3) or the Ion AmpliSeq ^™ Cancer Hotspot panel v2.

RNA-Seq gene expression analysis was performed using the AmpliSeq RNA analysis plugin in Torrent Suite Software. This plugin aligns raw sequence reads against the human reference genome (hg19 Ampliseq Transcriptome_ERCC_V1.fasta) containing 20,802 RefSeq transcripts using the Torrent Mapping Alignment Program (TMAP). The number of mapped reads per gene was then counted to generate a raw count file and the reads per gene were normalized to files per million mapped genes (RPM). . Tumor subtype assignments were made using the MD Anderson 1 nearest neighbor (oneNN) classifier. Eight tumors showed partial response (PR), four showed stable disease (SD), seven showed progressive disease (PD), and three were not classified (Figure 1).

To test for differential expression (DE) between pre- and post-treatment tumors, the Bioconductor package DESeq and the Negative Binomial model were used. The Benjamini-Hochberg method was used to control for false discovery rate (FDR). An unsupervised cluster analysis of baseline tissues revealed the presence of two clusters: Cluster 1 followed by Cluster 2 (2 PRs, 4 SDs, 6 PDs). were enriched for responders (6 PRs, 1 SD) compared to 1 (Fig. 1). Significantly differentially expressed genes from clusters 1 and 2 were extracted and analyzed by Ingenuity Pathway Analysis (Sigma).

To perform pathway analysis, published gene expression signature features of immune infiltration, inflammatory cell trafficking, IFN pathway activity or TH1 pathway activation, and FGFR3 gene expression signature were evaluated. Functional and pathway analyzes were performed using Ingenuity Pathway Analysis (IPA) software ( ^Ingenuity® Systems, Calif.), which contains a database for identifying networks and pathways of interest in genomic data. The "transcription factors as molecule type in upstream regulator" category within IPA was used to interpret the biological characteristics of bladder tumor subtypes. For upstream regulator analysis, IPA performs statistical analysis on overlap p-values and activation z-scores. Based on the IPA knowledge database, p-values and z-scores were calculated by matching how many targets for each transcription factor overlap (p-value) and the known effects (activation or inhibition) of targets in the gene list. (z-score).

The gene signatures used to identify basal and luminal subtypes were used against baseline tissue to assign 22 paired tumors to molecular subtypes (Fig. 2). Of the responding tumors, 6 were luminal and 2 were basal (Fig. 2); see also Denrauer 2014. Differential expression analysis in responders showed that bofatamab affected inflammatory and immune pathways and immune cell trafficking (Figure 3).

Collectively, these data demonstrate that luminal biology can sensitize subjects to combined treatment with bofatamab and pembrolizumab, regardless of the presence or absence of mutFGFR3. This suggests a role for this combination in wtFGFR3 mUC. The effect of the combination on tumors with basal biology also confirmed efficacy. These biological effects may also be important in early stage bladder cancer, such as non-muscle invasive bladder cancer (NMIBC). If bofatamab has an effect on immune cell trafficking, it may also show benefit in NMIBC, either after or in combination with BCG treatment.

Example 2: Clinical Evaluation of Bofatamab Monotherapy and Combination Therapy with Pembrolizumab in Subjects With mUC A FIERCE-22 Phase 1b/2 clinical trial was designed to evaluate bofatamab in combination with pembrolizumab. FIG. 4 shows a schematic diagram of the test. Subjects received bofatamab monotherapy for a two-week run-in period in pre-biopsy and post-biopsy pairings, followed by initiation of combination therapy. After the run-in period, combination therapy was initiated.

A first interim analysis was planned after enrollment of 26 patients. Twenty-eight patients were enrolled in the study. A second interim analysis is planned after enrollment of 52 patients. To be eligible for enrollment, mUC patients must be anti-PD-/L1 naive as well as measurable disease, ECOG greater than 2, and prior platinum-based chemotherapy on 1 or more lines. Progression or recurrence within 12 months of (neo)adjuvant chemotherapy was required. The demographics and treatment histories of the first 28 patients are shown in Table 1.
Table 1: Baseline population composition and treatment history:

＊ 貧血増悪および気管気管支炎 Primary endpoints were safety and efficacy. Overall response rate (ORR) was assessed by investigators using RECIST 1.1. Any biopsied lesion was considered non-target and began evaluation. The disposition of the first 28 patients is shown in Table 2. Median treatment exposure was extended with the majority of patients starting 8 or more treatment cycles.
Table 2: Properties - Phase 2:

* Exacerbation of anemia and tracheobronchitis

表４： 臨床応答－ＲＥＣＩＳＴ １．１ 評価可能集団：

＊ １名の患者は、ＰＲを確認する前に同意を撤回し、１名の患者は、次のスキャン時に偽性進行を有した。
¶ ｍｕｔ／ｆｕｓにおける患者は、追跡がおよそ４ヶ月短い。 The clinical responses of the first 28 patients in Phase 2 are shown in Table 3 and the clinical responses of the evaluable population from RECIST 1.1 are shown in Table 4.
Table 3: Clinical Response - Phase 2:

Table 4: Clinical Responses - RECIST 1.1 Evaluable Population:

* 1 patient withdrew consent before confirmation of PR and 1 patient had pseudoprogression at subsequent scan.
¶ Patients in mut/fus have approximately 4 months less follow-up.

＊ ８名のフェーズ１ｂ患者を含む。
¶ ４名のうちの１名は、フェーズ１ｂにおいて起こった。 The most common treatment-emergent adverse events (TEAEs) occurring in >15% or at least 15% of the 36 patients in Phases 1b and 2 are shown in Table 5. No cases of hyperphosphatemia or ocular or nail toxicity were reported. The majority of TEAEs, primarily Grade 1, occurred early on study and resolved during study treatment.
Table 5: TEAEs commonly occurring in ≧15% of Phase 1b/2 patients:

* Includes 8 Phase 1b patients.
¶ 1 of 4 occurred in Phase 1b.

＊ ８名のフェーズ１ｂ患者を含む。 The most common bofatamab-related TEAEs occurring in >15% or at least 15% of the 36 patients in Phases 1b and 2 are shown in Table 6. No bofatamab-related TEAEs of grade 3 or greater were reported. The most common bofatamab-associated TEAEs were fatigue and diarrhea.
Table 6: Bofatamab-related TEAEs in ≧15% of Phase 1b/2 patients

* Includes 8 Phase 1b patients.

The bofatamab lead-in period was well tolerated and paired biopsies were safely obtained from the majority (80%) of patients. Combination therapy with pembrolizumab was also well tolerated. No dose reductions of bofatamab occurred and interruptions were infrequent with 6 interruptions over 303 cycles. Serious AEs occurring in >2 patients were urinary tract infection (n=2; 6%) and acute kidney injury (n=2; 6%). All serious SEs were unrelated to bofatamab.

In the RECIST 1.1-Evaluable Phase 2 study population, the ORR was approximately double that of pembrolizumab alone. The ORR for all 22 patients with evaluable disease was 40% (9 of 22). The ORR for WT was 40% (6 out of 15) and 43% (3 out of 7) for Mut/Fus. Therefore, the response was similar regardless of FGFR3 Mut/Fus or WT status (Fig. 5, Fig. 13). Progression-free survival (PFS) trends appear to favor patients treated with combination therapy (5.9 months; 95% CI 3.6, 8.0) versus pembrolizumab alone (2.1 months; 95%) CI 2.0, 2.2). DOR is shorter than previously reported for CPI alone and does not appear to be an inference of OS. Median OS has not been achieved and all WT patients have been followed for at least 12 months of follow-up.

Bofatamab monotherapy during the run-in period induced inflammatory responses and immune alterations in responders, as well as upregulation of genes associated with immune pathway alterations (Fig. 6). Clinical responses correlated with molecular subgroups, with increased response rates (6 of 8) in the luminal subgroup (i.e., with tumors that were immunologically cold). subjects) were observed (FIGS. 7-12). This suggests that combination therapy may be particularly effective for this subtype. The presence of cancer-associated fibroblasts (CAFs) appears to be a possible mechanism associated with the lack of response. Patients with p53-like tumors appear to respond poorly (Figure 14).

Bofatamab produces immunological changes through FGFR3 inhibition. In a preliminary subtype study, the combination of bofatamab and pembrolizumab improves response in bladder cancer subtypes, with the exception of the p53-like group. Resistance to treatment may be associated with a p53-like subgroup of patients. Some patients had continued clinical benefit from treatment beyond RECIST 1.1 progression.
References

In certain embodiments, provided herein is the use of FGFR3 inhibitors and checkpoint inhibitors (eg, PD1 inhibitors) for use in the formulation of medicaments for the treatment of bladder cancer. In certain of these embodiments, the FGFR3 inhibitor and PD1 inhibitor are formulated into one medicament. In other embodiments, the FGFR3 inhibitor and PD1 inhibitor are formulated into separate medicaments that are administered in combination with each other.
The present invention provides, for example, the following items.
(Item 1)
1. A method of treating a luminal-type bladder cancer that expresses wild-type FGFR3 in a subject in need thereof, said method comprising administering to said subject a therapeutically effective amount of an FGFR3 inhibitor, a therapeutically effective amount of A method comprising administering in combination with a checkpoint inhibitor.
(Item 2)
2. The method of item 1, wherein said FGFR3 inhibitor is an antagonistic FGFR3 inhibitor.
(Item 3)
3. The method of item 2, wherein the antagonistic FGFR3 inhibitor is an antagonistic FGFR3 antibody.
(Item 4)
The antagonistic FGFR3 antibody comprises CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:1, CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO:2, and CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:3. 4. The method of item 3, comprising:
(Item 5)
5. The method of item 4, wherein said antagonistic FGFR3 antibody comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:7.
(Item 6)
The antagonistic FGFR3 antibody comprises CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:4, CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:5, and CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:6. 4. The method of item 3, comprising:
(Item 7)
7. The method of item 6, wherein said antagonistic FGFR3 antibody comprises a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:8.
(Item 8)
The method of item 1, wherein the FGFR3 inhibitor is bofatamab.
(Item 9)
The method of item 1, wherein the checkpoint inhibitor is a PD1 inhibitor.
(Item 10)
10. The method of item 9, wherein said PD1 inhibitor is an antagonistic PD-L1 antibody.
(Item 11)
11. The method of item 10, wherein said antagonistic PD-L1 antibody is selected from the group consisting of MEDI-4736, RG7446, BMS-936559, MSB0010718C, and MPDL3280A.
(Item 12)
10. The method of item 9, wherein the PD1 inhibitor is pembrolizumab.
(Item 13)
A method of treating a luminal-type bladder cancer expressing wild-type FGFR3 in a subject in need thereof, said method comprising combining a therapeutically effective amount of an antagonistic FGFR3 inhibitor with a therapeutically effective amount of a PD1 inhibitor. A method comprising administering in combination.
(Item 14)
14. The method of item 13, wherein said antagonistic FGFR3 inhibitor is an antagonistic FGFR3 antibody.
(Item 15)
The antagonistic FGFR3 antibody has a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 1, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 2, a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 3, and 15. The method of item 14, comprising a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO:7.
(Item 16)
The antagonistic FGFR3 antibody has a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:4, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:5, a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:6, and 15. The method of item 14, comprising a light chain variable region comprising the amino acid sequence shown in SEQ ID NO:8.
(Item 17)
14. The method of item 13, wherein said FGFR3 inhibitor is bofatamab.
(Item 18)
14. The method of item 13, wherein said PD1 inhibitor is an antagonistic PD-L1 antibody.
(Item 19)
19. The method of item 18, wherein said antagonistic PD-L1 antibody is selected from the group consisting of MEDI-4736, RG7446, BMS-936559, MSB0010718C, and MPDL3280A.
(Item 20)
14. The method of item 13, wherein the PD1 inhibitor is pembrolizumab.
(Item 21)
A method of treating a subject in need thereof having a cancer that expresses wild-type FGFR3, said method comprising:
(a) screening said subject for a genetic signature that correlates with one or more cancer-associated fibroblasts or for p53 expression;
(b) determining whether said subject has a genetic signature or has p53 expression that correlates with said one or more cancer-associated fibroblasts;
(c) based on the determination of step (b) above,
(i) if said subject does not have said gene signature or p53 expression, administering a therapeutically effective amount of an FGFR3 inhibitor in combination with a therapeutically effective amount of a checkpoint inhibitor; and
(ii) if said subject has said gene signature or p53 expression, administering a therapeutically effective amount of an FGFR3 inhibitor in combination with a therapeutically effective amount of a checkpoint inhibitor and an additional anti-cancer agent;
A method that encompasses
(Item 22)
22. The method of item 21, wherein said FGFR3 inhibitor is an antagonistic FGFR3 antibody.
(Item 23)
The antagonistic FGFR3 antibody has a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 1, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 2, a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 3, and 22. The method of item 21, comprising a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO:7.
(Item 24)
The antagonistic FGFR3 antibody has a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:4, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:5, a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:6, and 22. The method of item 21, comprising a light chain variable region comprising the amino acid sequence shown in SEQ ID NO:8.
(Item 25)
22. The method of item 21, wherein said FGFR3 inhibitor is bofatamab.
(Item 26)
22. The method of item 21, wherein the checkpoint inhibitor is a PD1 inhibitor.
(Item 27)
27. The method of item 26, wherein said PD1 inhibitor is an antagonistic PD-L1 antibody selected from the group consisting of MEDI-4736, RG7446, BMS-936559, MSB0010718C, and MPDL3280A.
(Item 28)
22. The method of item 21, wherein the PD1 inhibitor is pembrolizumab.
(Item 29)
22. The method of item 21, wherein the cancer is luminal bladder cancer.

Claims (29)

1. A method of treating a luminal-type bladder cancer that expresses wild-type FGFR3 in a subject in need thereof, said method comprising administering to said subject a therapeutically effective amount of an FGFR3 inhibitor, a therapeutically effective amount of A method comprising administering in combination with a checkpoint inhibitor. 2. The method of claim 1, wherein said FGFR3 inhibitor is an antagonistic FGFR3 inhibitor. 3. The method of claim 2, wherein said antagonistic FGFR3 inhibitor is an antagonistic FGFR3 antibody. The antagonistic FGFR3 antibody comprises CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:1, CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO:2, and CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:3. 4. The method of claim 3, comprising: 5. The method of claim 4, wherein said antagonistic FGFR3 antibody comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:7. The antagonistic FGFR3 antibody comprises CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:4, CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:5, and CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:6. 4. The method of claim 3, comprising: 7. The method of claim 6, wherein said antagonistic FGFR3 antibody comprises a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:8. 2. The method of claim 1, wherein the FGFR3 inhibitor is bofatamab. 2. The method of claim 1, wherein the checkpoint inhibitor is a PD1 inhibitor. 10. The method of claim 9, wherein said PD1 inhibitor is an antagonistic PD-L1 antibody. 11. The method of claim 10, wherein said antagonistic PD-Ll antibody is selected from the group consisting of MEDI-4736, RG7446, BMS-936559, MSB0010718C, and MPDL3280A. 10. The method of claim 9, wherein the PD1 inhibitor is pembrolizumab. A method of treating a luminal-type bladder cancer expressing wild-type FGFR3 in a subject in need thereof, said method comprising combining a therapeutically effective amount of an antagonistic FGFR3 inhibitor with a therapeutically effective amount of a PD1 inhibitor. A method comprising administering in combination. 14. The method of claim 13, wherein said antagonistic FGFR3 inhibitor is an antagonistic FGFR3 antibody. The antagonistic FGFR3 antibody has a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 1, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 2, a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 3, and 15. The method of claim 14, comprising a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:7. The antagonistic FGFR3 antibody has a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:4, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:5, a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:6, and 15. The method of claim 14, comprising a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:8. 14. The method of claim 13, wherein the FGFR3 inhibitor is bofatamab. 14. The method of claim 13, wherein said PD1 inhibitor is an antagonistic PD-L1 antibody. 19. The method of claim 18, wherein said antagonistic PD-Ll antibody is selected from the group consisting of MEDI-4736, RG7446, BMS-936559, MSB0010718C, and MPDL3280A. 14. The method of claim 13, wherein the PD1 inhibitor is pembrolizumab. A method of treating a subject in need thereof having a cancer that expresses wild-type FGFR3, said method comprising:
(a) screening said subject for a genetic signature that correlates with one or more cancer-associated fibroblasts or for p53 expression;
(b) determining whether said subject has a genetic signature or has p53 expression that correlates with said one or more cancer-associated fibroblasts;
(c) based on the determination of step (b) above,
(i) if said subject does not have said gene signature or p53 expression, administering a therapeutically effective amount of an FGFR3 inhibitor in combination with a therapeutically effective amount of a checkpoint inhibitor; and (ii) said subject. administering a therapeutically effective amount of an FGFR3 inhibitor in combination with a therapeutically effective amount of a checkpoint inhibitor and an additional anti-cancer agent when the body has said gene signature or p53 expression;
A method that encompasses 22. The method of claim 21, wherein said FGFR3 inhibitor is an antagonistic FGFR3 antibody. The antagonistic FGFR3 antibody has a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 1, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 2, a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 3, and 22. The method of claim 21, comprising a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:7. The antagonistic FGFR3 antibody has a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:4, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:5, a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:6, and 22. The method of claim 21, comprising a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:8. 22. The method of claim 21, wherein said FGFR3 inhibitor is bofatamab. 22. The method of claim 21, wherein said checkpoint inhibitor is a PD1 inhibitor. 27. The method of claim 26, wherein said PD1 inhibitor is an antagonistic PD-L1 antibody selected from the group consisting of MEDI-4736, RG7446, BMS-936559, MSB0010718C, and MPDL3280A. 22. The method of claim 21, wherein the PD1 inhibitor is pembrolizumab. 22. The method of claim 21, wherein the cancer is luminal bladder cancer.

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