CN113710244A

CN113710244A - Methods and compositions for treating cancer

Info

Publication number: CN113710244A
Application number: CN202080030176.5A
Authority: CN
Inventors: G.库里
Original assignee: Fusion Pharmaceuticals Inc
Current assignee: Fusion Pharmaceuticals Inc
Priority date: 2019-06-03
Filing date: 2020-03-03
Publication date: 2021-11-26
Also published as: CA3131880A1; KR20220016803A; JP2024144605A; BR112021017405A2; JP2023514795A

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. In some embodiments, the cancer expresses wild-type FGFR 3. The FGFR3 inhibitor can be an antagonistic FGFR3 inhibitor, such as an antagonistic FGFR3 antibody. The checkpoint inhibitor may be a PD1 inhibitor, including PD1 or PD1 ligand (PD-L1) antibodies, such as antagonistic PD1 or PD-L1 antibodies.

Description

Methods and compositions for treating cancer

Priority declaration

This application claims U.S. provisional patent application No. 62/812,929 filed on 3/1/2019; U.S. provisional patent application No. 62/856,216 filed on 3/6/2019; and U.S. provisional patent application No. 62/907,504, filed on 27.9.2019, the disclosure of which (including the figures) is incorporated herein by reference in its entirety.

Sequence listing

The present disclosure includes a sequence listing submitted in ASCII format over EFS-Web, which is incorporated herein by reference in its entirety. This ASCII copy was created on 3 months 3 of 2020 named sequence listing.

Background

Urothelial Cell Carcinoma (UCC), also known as Transitional Cell Carcinoma (TCC), occurs in the urinary system (i.e., kidney, bladder and appendages) and is the most common type of bladder cancer, accounting for 90% of all bladder tumors (Eble 2004). It is the fifth most common cancer in the United States (US) (costatini 2011) and the fourth most common cancer in europe (Jemal 2011); in 2014 74,690 new cases and 15,580 deaths were estimated to occur in US (American Cancer Society 2018), and in 2009 136,000 new cases and 49,000 deaths were estimated to occur in europe (Bellmunt 2009). Clinical and pathological studies have identified two variants of bladder urothelial cell carcinoma that occur by different mechanisms, low-grade papillary variants and invasive tumor variants (Wu 2005; Vallot 2010). The low-level papillary variants account for 80% of all UCBs and are derived from urothelial hyperplasia. The five-year survival of this tumor type is greater than 90% when treated with surgical and intravesical immunotherapy (American Cancer Society 2018). Aggressive tumor variants account for 20% of UCB and have a poorer prognosis. Cisplatin-based chemotherapy with dose-intensive M-VAC (Sternberg 2001) or gemcitabine cisplatin (von der Maase 2000) remains the standard treatment for aggressive UCC. Although the initial response rate is approximately 50% to 70%, this cancer usually progresses rapidly with a median survival of approximately 13 to 15 months (von der Maase 2000; Siefker-Radtke 2002). New therapies for UCC are needed.

Summary of The Invention

Provided herein are certain embodiments and methods of treating cancer, including, for example, bladder cancer, such as metastatic urothelial cancer (mUC) and upper urinary tract urothelial cancer, in a subject in need thereof, comprising administering a therapeutically effective amount of an FGFR3 inhibitor and a therapeutically effective amount of a checkpoint inhibitor, such as a PD1 inhibitor. In certain embodiments, the FGFR3 inhibitor binds to FGFR 3. In other embodiments, the FGFR3 inhibitor binds to a ligand of FGFR3, e.g., FGF1, FGF2, or FGF 9. In certain embodiments, the FGFR3 inhibitor is an antagonistic FGFR3 antibody, and in certain of these embodiments, the antagonistic FGFR3 antibody comprises one or more of: CDR-H1 comprising SEQ ID NO:1, CDR-H2 comprising SEQ ID NO:2, CDR-H3 comprising SEQ ID NO:3, heavy chain variable region comprising SEQ ID NO:7, heavy chain comprising SEQ ID NO:9, CDR-L1 comprising SEQ ID NO:4, CDR-L2 comprising SEQ ID NO:5, CDR-L3 comprising SEQ ID NO:6, light chain variable region comprising SEQ ID NO:8 and light chain comprising the amino acid sequence set forth in SEQ ID NO: 10. In certain of these embodiments, the FGFR3 antagonist antibody is vofatamab. 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 (pan-FGFR) inhibitor, and in certain of these embodiments the pan FGFR inhibitor is selected from the group consisting of inflixinib (infigratinib), AZD4547, LY2874455, pemitinib (pemitinib), BGJ398, logatinib (rogatatinib), PRN1371, Debio 1347, ARQ 087, and JNJ-42756493. In certain embodiments, the PD1 inhibitor binds to PD 1. In other embodiments, the PD1 inhibitor binds to a ligand of PD1, e.g., PDL1 or PDL 2. In certain embodiments, the PD1 inhibitor is an antagonistic PD1 antibody, and in certain of these embodiments, the antagonistic PD1 antibody is selected from the group consisting of nivolumab (nivolumab), pembrolizumab, CT-011, MEDI-0680, and RMP 1-14. In other embodiments, the PD1 inhibitor is an antagonistic PD1 ligand antibody, and in certain of these embodiments, the antagonistic PD1 ligand antibody is selected from the group consisting of MEDI-4736, RG7446, BMS-936559, MSB0010718C, and MPDL 3280A.

In certain embodiments, provided herein are compositions comprising an FGFR3 inhibitor and a checkpoint inhibitor, such as 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. In certain embodiments, the FGFR3 inhibitor binds to FGFR 3. In other embodiments, the FGFR3 inhibitor binds to a ligand of FGFR3, e.g., FGF1, FGF2, or FGF 9. In certain embodiments, the FGFR3 inhibitor is an antagonistic FGFR3 antibody, and in certain of these embodiments, the antagonistic FGFR3 antibody comprises one or more of: CDR-H1 comprising SEQ ID NO:1, CDR-H2 comprising SEQ ID NO:2, CDR-H3 comprising SEQ ID NO:3, heavy chain variable region comprising SEQ ID NO:7, heavy chain comprising SEQ ID NO:9, CDR-L1 comprising SEQ ID NO:4, CDR-L2 comprising SEQ ID NO:5, CDR-L3 comprising SEQ ID NO:6, light chain variable region comprising SEQ ID NO:8 and light chain comprising the amino acid sequence set forth in SEQ ID NO: 10. In certain of these embodiments, the FGFR3 antagonist antibody is vofatamab. 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 selected from the group consisting of inflixinib, AZD4547, LY2874455, pemitinib, BGJ398, mogertinib, PRN1371, Debio 1347, ARQ 087, and JNJ-42756493. In certain embodiments, the PD1 inhibitor binds to PD 1. In other embodiments, the PD1 inhibitor binds to a ligand of PD1, e.g., PDL1 or PDL 2. In certain embodiments, the PD1 inhibitor is an antagonistic PD1 antibody, and in certain of these embodiments, the antagonistic PD1 antibody is selected from the group consisting of nivolumab, pembrolizumab, CT-011, MEDI-0680, and RMP 1-14. In other embodiments, the PD1 inhibitor is an antagonistic PD1 ligand antibody, and in certain of these embodiments, the antagonistic PD1 ligand antibody is selected from the group consisting of MEDI-4736, RG7446, BMS-936559, MSB0010718C, and MPDL 3280A.

In certain embodiments, provided herein are methods of treating a cancer expressing wild-type FGFR3 in a subject in need thereof comprising administering to the subject a therapeutically effective amount of an FGFR3 inhibitor in combination with a therapeutically effective amount of a checkpoint inhibitor. In certain embodiments, the FGFR3 inhibitor binds to FGFR 3. In other embodiments, the FGFR3 inhibitor binds to a ligand of FGFR3, e.g., FGF1, FGF2, or FGF 9. In certain embodiments, the FGFR3 inhibitor is an antagonistic FGFR3 antibody, and in certain of these embodiments, the antagonistic FGFR3 antibody comprises one or more of: CDR-H1 comprising SEQ ID NO:1, CDR-H2 comprising SEQ ID NO:2, CDR-H3 comprising SEQ ID NO:3, heavy chain variable region comprising SEQ ID NO:7, heavy chain comprising SEQ ID NO:9, CDR-L1 comprising SEQ ID NO:4, CDR-L2 comprising SEQ ID NO:5, CDR-L3 comprising SEQ ID NO:6, light chain variable region comprising SEQ ID NO:8 and light chain comprising the amino acid sequence set forth in SEQ ID NO: 10. In certain of these embodiments, the FGFR3 antagonist antibody is vofatamab. 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 selected from the group consisting of inflixinib, AZD4547, LY2874455, pemitinib, BGJ398, mogertinib, PRN1371, Debio 1347, ARQ 087, and JNJ-42756493. In certain embodiments, the checkpoint inhibitor is a PD1 inhibitor. In certain of these embodiments, the inhibitor of PD1 binds to PD 1. In other embodiments, the PD1 inhibitor binds to a ligand of PD1, e.g., PDL1 or PDL 2. In certain embodiments, the PD1 inhibitor is an antagonistic PD1 antibody, and in certain of these embodiments, the antagonistic PD1 antibody is selected from the group consisting of nivolumab, pembrolizumab, CT-011, MEDI-0680, and RMP 1-14. In other embodiments, the PD1 inhibitor is an antagonistic PD1 ligand antibody, and in certain of these embodiments, the antagonistic PD1 ligand antibody is selected from the group consisting of MEDI-4736, RG7446, BMS-936559, MSB0010718C, and MPDL 3280A.

In certain embodiments, provided herein are methods of treating a subject in need thereof with a cancer that expresses wild-type FGFR3 comprising (a) screening the subject for a gene signature associated with one or more cancer-associated fibroblasts, or for p53 expression, (b) determining whether the subject has a gene signature associated with one or more cancer-associated fibroblasts or has p53 expression, (c) administering a therapeutically effective amount of an FGFR3 inhibitor in combination with a therapeutically effective amount of a checkpoint inhibitor based on the determination of step (b) and (i) if the subject does not have the gene signature or p53 expression, and (ii) if the subject has the 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. In certain embodiments, the FGFR3 inhibitor is an antagonistic FGFR3 antibody. In certain embodiments, 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, CDR-H3 comprising the amino acid sequence set forth in SEQ ID No. 3, and a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID No. 7. In certain embodiments, 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, CDR-L3 comprising the amino acid sequence set forth in SEQ ID No. 6, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID No. 8. In certain embodiments, the FGFR3 inhibitor is vofatamab. In certain embodiments, the checkpoint inhibitor is a PD1 inhibitor. In certain embodiments, the PD1 inhibitor is an antagonistic PD-L1 antibody selected from the group consisting of MEDI-4736, RG7446, BMS-936559, MSB0010718C, and MPDL 3280A. In certain embodiments, the PD1 inhibitor is pembrolizumab. In certain embodiments, the cancer is luminal bladder cancer. In certain embodiments, the gene signature includes at least the following genes: FGFR3, TP63, IRS1, SEMA4B, PTPN13 and TMPRSS 4. In certain embodiments, the gene signature includes at least the following genes: KRT5, KRT6A, KRT6B, KRT14, UPK3A, UPK3B, FOXA1 and PPARG. In certain embodiments, the gene signature includes at least the following genes: ACTC1, ACTG2, NCC1, DES, FLNC, MFAP4, MYH11, and PCP 4.

In certain embodiments, provided herein are kits comprising an FGFR3 inhibitor and a checkpoint inhibitor, such as a PD1 inhibitor, 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 an FGFR3 inhibitor and a checkpoint inhibitor, such as a PD1 inhibitor, for the formulation of a medicament for the treatment of bladder cancer. In certain of these embodiments, the FGFR3 inhibitor and the PD1 inhibitor are formulated as a single agent. In other embodiments, the FGFR3 inhibitor and the PD1 inhibitor are formulated as separate medicaments to be administered in combination with each other.

Brief Description of Drawings

Figure 1 depicts the results of whole transcriptome RNAseq performed on 22 matched tumor biopsies illustrating two prognostic clusters before and after vofatamab dosing. Cluster 1 is predominantly lumen in type and includes the majority of responders (respondents). Cluster 2 is substantially more basal and shows clinical benefit consistent with expected response rates.

Figure 2 depicts the results of whole transcriptome RNAseq performed on 22 matched tumor biopsies. 22 paired tumors were assigned to molecular subtypes using gene signatures for identification of basal and luminal subtypes.

Figure 3 shows the changes in inflammatory and immune pathways in responders following vofatamab treatment.

Figure 4 is a schematic of phase 1 b/phase 2 study design examining the effect of vofatamab plus pembrolizumab at second line mUC.

FIG. 5 shows that patient responsiveness can be assessed based on RECIST1.1 in a phase 2 study of WT versus Mut/Fus.

Figure 6 shows the immune changes in responders.

FIG. 7 is a spider graph depicting the change from baseline in the sum of diameters (SoD) of lesions after vofatamab (B-701) treatment. The plot shows that vofatamab appears to induce an initial significant increase in tumor size followed by a dramatic decrease in tumor volume.

FIG. 8 illustrates an emerging definition of molecular subtypes for bladder cancer.

Figure 9 shows that RECIST1.1 can assess patient responsiveness in phase 2 studies based on molecular subgroups.

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

Figure 11 shows the duration of treatment in phase 2 studies by molecular subgroup.

Figure 12 shows the results of combined vofatamab and pembrolizumab treatment in a luminal subtype of male mUC patient.

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

Figure 14 shows survival after treatment with combination vofatamab and pembrolizumab. Patients with p 53-like tumors (n ═ 5) exhibited poor survival.

Detailed Description

The following description of the invention is intended to be illustrative of various embodiments of the invention only. 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, changes, and modifications may be made without departing from the scope of the invention, and it is to be understood that such equivalent embodiments are to be included herein.

There are four single transmembrane tyrosine kinase fibroblast growth factor receptors (FGFR1-4) in humans (Brooks 2012). FGFR is overexpressed in many cancer types, largely due to mutations that confer constitutive activation, making them attractive targets for therapeutic intervention. For example, the FGFR2 antibody FPA144 (veprime) is currently being developed 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-FGFR 4 has also been reported to inhibit tumor growth (Bumbaca 2011).

FGFR3 carries both tumorigenic and tumor-inhibiting properties. FGFR3 is frequently mutated in certain cancers, but in some normal tissues it can limit cell growth and promote cell differentiation (Lafitte 2013). The human FGFR3 antagonistic monoclonal antibody, vofatamab (sometimes referred to as B-701 or BM2), was the first FGFR3 targeting antibody to enter clinical development. The variable portion of vofatamab was initially identified by phage display and subsequently recombined with the human IgG1 backbone. Votatamab binds wild-type and mutant FGFR3 with high affinityBoth, including the most common mutations found in bladder cancer and achondroplasia (specifically, FGFR 3-IIIb)^R248C、FGFR3-IIIb^K652E、FGFR3-III^Y375C、FGFR3-IIIb^S249CAnd FGFR3-IIIb^G372C) And gene fusions (including FGFR3 TACC3 and FGFR3 BAIAP2L1), but did not exhibit cross-reactivity with other FGFR 3. The safety of Vofatamab in patients with t (4:14) translocating multiple myeloma was previously assessed (Clinical Trial NCT 01122875). Other FGFR3 inhibitor antibodies that have been developed clinically or preclinically include PRO-001(Prochon), IMC-D11(Imclone), and ADC LY3076226(Eli Lilly). Additional FGFR3 antibodies for the treatment of cancer and other diseases have been disclosed in, for example, U.S. patent nos. 8,187,601(Aveo) and 7,498,416 (fibrin).

Programmed cell death protein 1(PD1) is an immune checkpoint receptor from the CD28 superfamily that restricts T cell effector function within tissues upon activation by one of its two ligands PDL1 or PDL2 (pardol 2012). PD1 down-regulates the immune system by promoting apoptosis in antigen-specific T cells while reducing apoptosis in regulatory (i.e., inhibitory) T cells. Certain tumor cells block anti-tumor immune responses in the tumor microenvironment by upregulating the ligand of PD 1. Blocking the PD1 pathway activates the immune system to attack tumors and has been shown to 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: (

Bristol-Myers Squibb and Ono Pharmaceutical; also known as ONO-4538, BMS-936558, MDX-1106) has been approved for the treatment of metastatic melanoma patients who have no alternative or no longer responded to other drugs. Nivolumab is also being evaluated in combination with various chemotherapeutic regimens for the treatment of non-small cell lung cancer (NSCLC). Humanized IgG4 PD1 antibody pembrolizumab (C)

Merck; also known as MK-3475) has been approved for several cancersTypes of disorders, including treatment of UCC, NSCLC and melanoma. Other PD1 antibodies under development include CT-011(Curetech) and MEDI-0680/AMP-514 (AstraZeneca).

A variety of PD1 ligand (PDL) antibodies are also being developed for cancer therapy. For example, monoclonal IgG1k PDL1 antibody MEDI-4736(AstraZeneca), alone or in combination with the monoclonal CTLA4 antibody tremelimumab (tremelimumab) (AstraZeneca) or MEDI-0680 is currently being developed for treatment of NSCLC, monoclonal IgG1k PDL1 antibody RG7446(Roche), alone or in combination

And

a fully human monoclonal IgG4 antibody BMS-936559/MDX-1105(BMS) is currently being developed for the treatment of various cancers, a fully human IgG1 PDL1 antibody MSB0010718C (Merck Serono) is currently being developed for the treatment of various cancer types, and an Fc modified monoclonal IgG4 antibody MPDL3280A (Genentech) is currently being developed for the treatment of NSCLC.

As shown in the experimental results provided herein, the FGFR3 inhibitor, vofatamab, has been shown to induce activation of genes associated with TH1 responses and key signaling molecules in immune function. These results indicate that vofatamab activates genes associated with the immune cell trafficking pathway. The results also indicate that vofatamab sensitizes patients with mUC with luminal biology to combination therapy with vofatamab and checkpoint inhibitors, e.g., PD1 inhibitors. The results provided herein further show that the gene signature associated with cancer-associated fibroblasts appears to be associated with resistance to the combination of vofatamab and checkpoint inhibitors, and that patients with p 53-like tumors also exhibit resistance to combination therapy. This suggests that patients exhibiting a cancer-associated fibroblast gene signature or having a p 53-like tumor may be candidates for combination therapy further comprising a third agent, e.g., an anti-fibrotic agent.

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. Also provided herein are methods of increasing the effectiveness of a PD1 inhibitor for treating cancer in a subject in need thereof, comprising administering an FGFR3 inhibitor, and methods of increasing the effectiveness of an FGFR3 inhibitor for treating cancer in a subject in need thereof, comprising administering a PD1 inhibitor. An increase in the effectiveness of a PD1 or FGFR3 inhibitor can refer to an increase in the therapeutic effect of either inhibitor, a decrease in the dosage required to achieve a certain level of therapeutic effect, the frequency of administration, or the time interval over which either inhibitor is administered, or some combination thereof.

In certain embodiments, the methods provided herein are used to treat solid cancers, i.e., cancers that form discrete tumor masses. In certain of these embodiments, the cancer being treated is bladder cancer, including, for example, metastatic bladder cancer (mUC) or upper urinary tract urothelial cancer.

As used herein, with respect to solid cancers, the term "treatment" may refer to partial or total inhibition of tumor growth, reduction in tumor size, complete or partial eradication of a tumor, reduction or prevention of malignant growth, partial or total eradication of cancer cells, or some combination thereof. The terms "patient" and "subject" are used interchangeably herein.

As used herein, "subject in need thereof" refers to a mammalian subject, preferably a human, that has been diagnosed as having cancer, is suspected of having cancer, and/or exhibits one or more symptoms associated with cancer. In certain embodiments, the subject may have previously received one or more therapeutic interventions for cancer treatment, e.g., chemotherapy.

As used herein, an "FGFR 3 inhibitor" refers to any molecule that partially or completely inhibits FGFR3 activity. An FGFR3 inhibitor may specifically inhibit FGFR3, or it may inhibit the activity of other proteins than FGFR 3. For example, FGFR3 inhibitors can also inhibit the activity of other FGFR 3. In certain embodiments, the FGFR3 inhibitor can exhibit indirect immunomodulatory activity. For example, FGFR3 inhibitors may have an indirect effect on immune cell trafficking pathways, e.g., by altering the expression and/or activity of TNF alpha or IFN gamma.

As used herein, "antagonist antibody" refers to an antibody that reduces, prevents, or otherwise inhibits the interaction between a receptor and its cognate ligand by physically binding to the receptor or its cognate ligand at a binding site and/or an allosteric site on either molecule. In some embodiments, the antagonist antibody interacts at a unique binding site that is not otherwise involved in the biological modulation of receptor activity by a cognate ligand. Thus, an antagonist antibody has affinity for a receptor or a cognate ligand, but has no efficacy in promoting a biological response upon binding as compared to a cognate ligand that binds to a receptor. Thus, the binding of the antagonist antibody disrupts the interaction of the receptor and the cognate ligand, and otherwise inhibits the function of the agonist.

In certain embodiments of the methods, compositions, and kits provided herein, the FGFR3 inhibitor inhibits FGFR3 activity by binding to FGFR 3. Examples of such FGFR3 inhibitors include, for example, antagonistic FGFR3 inhibitors or fusion proteins thereof, inactive forms of 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, meaning that the inhibitor exhibits little or no binding to other FGFRs. In other embodiments, the FGFR3 inhibitor binds one or more FGFR in addition to FGFR 3.

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 inhibitor specifically binds to FGFR 3. As used herein, the term "antibody" refers to an immunologically active portion of an immunoglobulin molecule or binding specific antigen (e.g., FGFR3 or PD 1). In those embodiments where the antibody used in the present methods, compositions, and kits is a full-length immunoglobulin molecule, the antibody comprises two heavy chains and two light chains, each heavy and light chain comprising three Complementarity Determining Regions (CDRs). In those embodiments in which the antibody is an immunologically active portion of an immunoglobulin, the antibody can be, for example, a Fab, Fab ', Fv, Fab ' F (ab ')2, disulfide-linked Fv, scFv, single domain antibody (dAb), or diabody (diabody). The antibodies used in the present methods, compositions, and kits may include natural antibodies, synthetic antibodies, monoclonal antibodies, polyclonal antibodies, chimeric antibodies, humanized antibodies, multispecific antibodies, bispecific antibodies, dual specific antibodies, anti-idiotypic antibodies, or fragments thereof that retain the ability to bind a specific antigen (e.g., FGFR3 or PD 1). 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 used in the present methods, compositions, and kits comprise a heavy chain variable region comprising 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 set forth in SEQ ID No. 4. In certain embodiments, the FGFR3 antagonist antibody comprises a light chain variable region comprising one or more CDRs having a sequence 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 set forth 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 comprises the heavy chain variable region of SEQ ID No. 4 and the light chain variable region of SEQ ID No. 8. In certain embodiments, the antibody is a vofamomab comprising the heavy chain of SEQ ID NO. 9 and the light chain of SEQ ID NO. 10. In addition to the variable region shown in SEQ ID NO 7, heavy chain SEQ ID NO 9 comprises human IgG 1. Similarly, the light chain of SEQ ID NO 10 comprises the variable region shown in SEQ ID NO 8 and human Ig kappa 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:

EVQLVESGGGLVQPGGSLRLSCAASGFTFTSTGISWVRQAPGKGLEWVGRIYPTSGSTNYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARTYGIYDLYVDYTEYVMDYWGQGTLV。

SEQ ID NO:8:

DIQMTQSPSSLSASVGDRVTITCRASQDVDTSLAWYKQKPGKAPKLLIYSASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSTGHPQTFGQGTKVEIKR。

SEQ ID NO:9:

EVQLVESGGGLVQPGGSLRLSCAASGFTFTSTGISWVRQAPGKGLEWVGRIYPTSGSTNYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARTYGIYDLYVDYTEYVMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK。

SEQ ID NO.10:

DIQMTQSPSSLSASVGDRVTITCRASQDVDTSLAWYKQKPGKAPKLLIYSASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSTGHPQTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC。

In other embodiments, the FGFR3 antagonist antibody used in the present methods, compositions, and kits can be a PRO-001, IMC-D11, or FGFR3 antagonist antibody, as disclosed in U.S. patent No. 8,187,601(Aveo) or 7,498,416 (fibrin). In certain embodiments, the FGFR3 antagonist antibodies used in the present methods, compositions, and kits 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, e.g., FGF1, FGF2, or FGF 9. Examples of such FGFR3 inhibitors include, for example, antibodies or fusion proteins thereof that specifically bind FGFR3 ligands, soluble forms of FGFR3 or fusion proteins thereof comprising all or part of the extracellular domain of FGFR3, truncated forms of FGFR3 or fusion proteins thereof lacking all or part of the intracellular domain required for downstream signaling, small molecules, sirnas, and aptamers.

In certain embodiments of the methods, compositions, and kits provided herein, the FGFR3 inhibitor is a pan FGFR inhibitor, meaning that it binds to one or more FGFRs other than FGFR3 and inhibits its activity. In certain of these embodiments, the FGFR3 inhibitor can be a small molecule pan FGFR inhibitor selected from the group consisting of infliximab (BGJ398, Novartis), AZD4547(AstraZeneca), LY2874455(Eli Lilly), Debio 1347(Debiopharm), ARQ 087 (arqual), 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 dovirtinib, lucitinib, ponatinib, nintedanib, ponatinib or emmd-2076 may be used as FGFR3 inhibitors.

As used herein, a "PD 1 inhibitor" refers to a molecule that partially or completely inhibits the activity of PD 1. A PD1 inhibitor may specifically inhibit PD1, or it may inhibit the activity of other proteins than PD 1. 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 PD 1. Examples of such PD1 inhibitors include, for example, antagonistic PD1 antibodies or fusion proteins thereof, inactive forms of PD1 ligand (e.g., truncated or other mutated forms of PDL1 or PDL2) or fusion proteins thereof (e.g., AMP-224(GlaxoSmithKline, amplimune)), 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 antagonizes the antibody's specific binding to PD 1. In certain embodiments, the PD1 antagonist antibody is selected from the group consisting of nivolumab, pembrolizumab, CT-011, MEDI-0680, and RMP 1-14.

In certain embodiments of the methods, compositions, and kits provided herein, the PD1 inhibitor inhibits PD1 activity by binding to one or more ligands of PD1 (i.e., PDL1 or PDL 2). Examples of such PD1 inhibitors include, for example, a PD1 ligand antibody or fusion protein thereof, a soluble form of PD1 comprising all or a portion of the extracellular domain of PD1 or a fusion protein thereof, a truncated form of PD1 lacking all or a portion of the intracellular domain required for downstream signaling or a fusion protein 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 to a PD1 ligand. In certain embodiments, the PD1 ligand antibody is selected from the group consisting of MEDI-4736, RG7446, BMS-936559, MSB0010718C, and MPDL 3280A.

In certain embodiments of the methods provided herein, the FGFR3 inhibitor and the PD1 inhibitor are administered together as part of the same composition. In other embodiments, the FGFR3 inhibitor and the PD1 inhibitor are administered separately, i.e., as separate compositions. In these embodiments, the inhibitors may be administered simultaneously or sequentially, and may be administered via the same or different routes. In those embodiments where the inhibitors are administered sequentially, 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 time course. In certain of these embodiments, one inhibitor may be administered one or more times prior to the first administration of a second inhibitor. When administration of the second inhibitor is initiated, administration of the first inhibitor may be stopped or continued over the course of all or part of the administration of the second inhibitor. In certain embodiments wherein the FGFR3 inhibitor is an FGFR3 antagonist antibody, the antibody can be administered twice or more daily, once daily, twice or more weekly, biweekly (i.e., every other week), every three weeks, or monthly. 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 can be administered twice or more daily, once daily, twice or more weekly, biweekly, triweekly, or monthly. In certain embodiments, the PD1 inhibitor is administered once every two weeks. In certain embodiments, the FGFR3 inhibitor and/or the PD1 inhibitor can be administered for a predetermined specific time course. For example, the FGFR3 and/or PD1 inhibitor can be administered over a time 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 reached. For example, FGFR3 and/or PD1 inhibitors can be administered until tumor growth is arrested or reversed, until one or more tumors are eliminated, or until the number of cancer cells is reduced to a particular level.

As used herein, a "therapeutically effective amount" of a composition is the amount of the composition that produces a desired therapeutic effect in a subject, such as treating cancer. In certain embodiments, a therapeutically effective amount is the amount of the composition that produces the greatest therapeutic effect. In other embodiments, the therapeutically effective amount produces a therapeutic effect that is less than the maximum therapeutic effect. For example, a therapeutically effective amount can be an amount that produces a therapeutic effect while avoiding one or more side effects associated with the dose that produces the greatest therapeutic effect. The therapeutically effective amount of a particular composition will vary based on a variety of factors including, but not limited to, the characteristics of the therapeutic composition (e.g., activity, pharmacokinetics, pharmacodynamics, and bioavailability), the physiological condition of the subject (e.g., age, body weight, sex, type and stage of disease, medical history, general physical condition, response to a given dose, and other existing drug treatments), the nature of any pharmaceutically acceptable carrier in the composition, and the route of administration. One skilled in the clinical and pharmacological arts will be able to determine a therapeutically effective amount by routine experimentation, i.e., by monitoring the subject's response to administration of the composition and adjusting the dosage accordingly. For more guidance, see, e.g., Remington, The Science and Practice of Pharmacy, 22 nd edition, Pharmaceutical Press, London, 2012 and Goodman & Gilman's The Pharmaceutical Basis of Therapeutics, 12 th edition, McGraw-Hill, New York, NY, 2011, The complete disclosure of which is incorporated herein by reference.

In certain embodiments of the methods provided herein, the therapeutically effective amount of the FGFR3 inhibitor or PD1 inhibitor can be a dose at which the molecule is capable of producing a therapeutic response (e.g., reducing or eliminating tumor growth) as a monotherapy, i.e., when administered alone. In certain of these embodiments, the therapeutically effective amount may be a dose that has been previously determined to be optimal or near optimal for the treatment of cancer. For example, where the FGFR3 inhibitor is vofatamab, the antibody can be administered at a dose of about 10 to 50mg/kg every two to four weeks, and in certain of these embodiments, the antibody can be administered at a dose of about 20 to 40mg/kg every two to four weeks, or about 30mg/kg every three weeks. In other embodiments, the therapeutically effective amount of the FGFR3 inhibitor or the PD1 inhibitor can be lower than the dose at which the molecule is typically administered as monotherapy, i.e., a sub-optimal dose. In certain of these embodiments, administration of a suboptimal dose of an FGFR3 or PD1 inhibitor may result in reduced side effects compared to the standard dose when administered alone. For example, administration of suboptimal doses of an FGFR3 or PD1 inhibitor may result in reduced occurrence or severity of pruritus, colitis, or pneumonia compared to administration of optimal doses of either inhibitor alone. In certain embodiments, one of the FGFR3 inhibitor and the PD1 inhibitor can be administered at a dose that has been determined to be optimal for cancer treatment when administered alone, while the other is administered at a dose that is suboptimal for treatment when administered alone. In certain embodiments, the dosage of the FGFR3 inhibitor or the PD1 inhibitor can be varied over the course of a treatment regimen. For example, one or both of the FGFR3 inhibitor and the PD1 inhibitor can be administered at a higher dose at the beginning of treatment (e.g., loading phase) followed by a lower dose at a later stage of treatment.

The FGFR3 inhibitor, PD1 inhibitor, or composition comprising both the FGFR3 inhibitor and the PD1 inhibitor can be delivered to a subject by any route of administration known in the art, including, but not limited to, parenteral, oral, aerosol, enteral, nasal, ophthalmic, parenteral, or transdermal (e.g., topical creams or ointments, patches). "parenteral" refers to a route of administration typically associated with injection, including intravenous, intraperitoneal, subcutaneous, infraorbital, infusion, intraarterial, intracapsular, intracardiac, intradermal, intramuscular, intrapulmonary, intraspinal, intrasternal, intrathecal, intrauterine, subarachnoid, subcapsular, transmucosal, or transtracheal. In certain embodiments wherein the FGFR3 inhibitor is a FGFR3 antagonist antibody, including, for example, vofatamab, 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, the FGFR3 inhibitor, the PD1 inhibitor, or the composition comprising both the FGFR3 and the PD1 inhibitor can be formulated as an oral dosage unit, such as, for example, a tablet, pill, or capsule. In certain embodiments, the FGFR3 inhibitor, the PD1 inhibitor, or the FGFR3 and PD1 inhibitor compositions can be administered via a time-release delivery vehicle, such as, for example, a time-release capsule. As used herein, "time release vehicle" refers to any delivery vehicle that releases an active agent over a period of time after administration, rather than immediately. In other embodiments, the FGFR3 inhibitor, the PD1 inhibitor, or the 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 may receive additional therapy, including, for example, chemotherapy or immunotherapy, or a third agent such as an anti-fibrotic agent, before, during, or after treatment with the FGFR3 and PD1 inhibitors. In those embodiments in which the subject receives additional therapy during FGFR3 and PD1 inhibitor treatment, the additional therapy may be administered simultaneously or sequentially with the FGFR3 inhibitor and/or the PD1 inhibitor.

In certain embodiments, provided herein are compositions comprising a therapeutically effective amount of an FGFR3 inhibitor and a therapeutically effective amount of a PD1 inhibitor. In certain embodiments, these compositions further comprise, or are formulated for administration with, one or more pharmaceutically acceptable carriers. Also provided herein are kits comprising an FGFR3 inhibitor and a PD1 inhibitor for performing the methods disclosed herein, e.g., for treating cancer.

In certain embodiments of the compositions and kits provided herein, the FGFR3 inhibitor or PDl inhibitor can be present in the composition or kit at a dose that is capable of producing a therapeutic response (e.g., reducing or eliminating tumor growth) when administered alone. In certain of these embodiments, the FGFR3 or PD1 inhibitor can be present at a dosage that has been previously determined to be optimal or near optimal for cancer treatment. For example, where the FGFR3 inhibitor is vofatamab, the composition or kit can be formulated to deliver a dose of about 10 to 50mg/kg of vofatamab to the subject, and in certain of these embodiments, the composition or kit can be formulated to deliver a dose of about 20 to 40mg/kg or about 30mg/kg of vofatamab to the subject. In other embodiments, the FGFR3 or PD1 inhibitor can be present at a dose (i.e., sub-optimal dose) that is lower than it is typically present in a composition or kit for cancer treatment.

As used herein, "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition, or vehicle involved in carrying or transporting a compound or molecule of interest from one tissue, organ, or body part to another. A pharmaceutically acceptable carrier may comprise various components including, but not limited to, liquid or solid fillers, diluents, excipients, solvents, buffers, encapsulating materials, surfactants, stabilizers, binders, or pigments, or some combination thereof. Each component of the carrier must be "pharmaceutically acceptable" in that it must be compatible with the other ingredients of the composition and must be suitable for contact with any tissue, organ, or body part with which it may be encountered, meaning that it must not carry the risk of toxicity, irritation, allergic response, immunogenicity, or any other complications that outweigh their therapeutic benefits.

Examples of pharmaceutically acceptable carriers that can be used in conjunction with the compositions provided herein include, but are not limited to, (1) sugars such as lactose, glucose, sucrose, or mannitol; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives such as carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (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 oil; (10) glycols such as propylene glycol; (11) polyols such as glycerol, sorbitol, mannitol and polyethylene glycol; (12) esters such as ethyl oleate and ethyl laurate; (13) disintegrating agents such as agar or calcium carbonate; (14) buffers or pH adjusters 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 ethanol and propanol; (20) paraffin wax; (21) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol or sodium lauryl sulfate; (22) a colorant or pigment; (23) glidants such as colloidal silicon dioxide, talc and starch or tricalcium phosphate; (24) other non-toxic compatible substances used in pharmaceutical compositions, such as acetone; and (25) combinations thereof.

The compositions comprising the FGFR3 inhibitor, the PD1 inhibitor, or the FGFR3 inhibitor and the PD1 inhibitor in combination can be formulated into suitable dosage forms including, for example, solutions or suspensions in aqueous or non-aqueous liquids, oil-in-water or water-in-oil liquid emulsions, capsules, cachets, pills, tablets, lozenges, powders, granules, elixirs or syrups, or lozenges. In certain embodiments, the compositions may be formulated as a time-release delivery vehicle, such as, for example, a time-release capsule. As used herein, "time release vehicle" refers to any delivery vehicle that releases an active agent over a period of time after administration, rather than immediately. In other embodiments, the compositions may be formulated as an immediate release delivery vehicle.

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

In certain embodiments, provided herein are FGFR3 inhibitors and PD1 inhibitors for use in the treatment of cancer. Also provided are FGFR3 inhibitors for use in the treatment of cancer, and PD1 inhibitors for use in the treatment of cancer in combination with FGFR3 inhibitors. In certain of these embodiments, the cancer is a urothelial cancer, such as 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 gene signature may be a candidate for treatment with an FGFR3 inhibitor and a PD1 inhibitor of the present disclosure. As another example, a subject in need of treatment for cancer with another specific gene signature may not be a candidate for treatment with an FGFR3 inhibitor and a PD1 inhibitor of the present disclosure, but may have treatment comprising a third agent of the present disclosure. In some embodiments, another specific gene signature comprises a gene signature associated with cancer-associated fibroblasts and a p 53-like tumor. In certain embodiments, the gene signature is an FGFR3 gene signature and includes at least the following genes: FGFR3, TP63, IRS1, SEMA4B, PTPN13 and TMPRSS 4. In certain embodiments, the gene signature is a p 53-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 PCP 4.

In certain embodiments, provided herein are methods for treating a subject in need thereof with a cancer that expresses wild-type FGFR3 comprising (a) screening the subject for a gene signature associated with one or more cancer-associated fibroblasts or for p53 expression, (b) determining whether the subject has a gene signature associated with one or more cancer-associated fibroblasts or has p53 expression, (c) administering a therapeutically effective amount of an FGFR3 checkpoint inhibitor in combination with a therapeutically effective amount of an anti-cancer agent based on the determination of step (b) and (i) if the subject does not have the gene signature or p53 expression, and (ii) if the subject has the gene signature or p53 expression, administering a therapeutically effective amount of an FGFR3 inhibitor in combination with a therapeutically effective amount of the 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 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, CDR-H3 comprising the amino acid sequence set forth in SEQ ID No. 3, and a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID No. 7. In certain embodiments, 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, CDR-L3 comprising the amino acid sequence set forth in SEQ ID No. 6, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID No. 8. In certain embodiments, the FGFR3 inhibitor is vofatamab. In certain embodiments, the checkpoint inhibitor is a PD1 inhibitor. In certain embodiments, the PD1 inhibitor is an antagonistic PD-L1 antibody selected from the group consisting of MEDI-4736, RG7446, BMS-936559, MSB0010718C, and MPDL 3280A. 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 an FGFR3 inhibitor and a PD1 inhibitor in the manufacture of a medicament for the treatment of cancer. Also provided is the use of an FGFR3 inhibitor for the manufacture of a medicament for the treatment of cancer in combination with a PD1 inhibitor, and the use of a PD1 inhibitor for the manufacture of a medicament for the treatment of cancer in combination with an FGFR3 inhibitor.

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

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

From the foregoing, it will be appreciated that specific embodiments of the invention have been described herein for purposes of illustration, but that various modifications may be made without deviating 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 that specific materials are mentioned, they are for illustrative purposes only and are not intended to limit the invention. Those skilled in the art can develop equivalent means or reactants without losing the ability of the invention and without departing from the scope of the invention. It will be appreciated that many variations in the steps described herein may be made while still remaining within the bounds of the present invention. It is the intention of the inventors that such variations are included within the scope of the invention.

Examples

Example 1: wild type and mutant FGFR3 treated with combination therapy with vofatamab and pembrolizumab Gene expression profile of metastatic urothelial cancer

In patients with metastatic urothelial cancer (mUC), the FGFR3 mutation (mutFGFR3) is present in 15-20% of bladder cancer patients and around 35% of upper urinary tract cancer patients. Studies indicate that these tumors may respond better to FGFR inhibition than immunotherapy.

Patients who failed prior therapy mUC were treated with vofatamab. Patients with pre-treatment and 14 days post-treatment biopsies were treated with a loading dose of vofatamab (25mg/kg), followed by a cohort of patients with and without mutFGFR3 or gene infusion (gene infusion) with pembrolizumab combination therapy. Core biopsies (2x18G or 3-4x20G) were fixed in 10% neutral buffered formalin and embedded in paraffin. The biopsy core was then isolated from paraffin blocks of these Formalin Fixed Paraffin Embedded (FFPE) samples.

DNA and RNA were isolated from the samples for whole transcriptome RNAseq and DNA sequencing on Ion Torrent platform. Samples from 22 patients (17 WT, 5 Mut fusions) with matched pre-and post-treatment biopsies were subjected to whole transcriptome RNAseq using the AmpliSeqRNA platform from Ion Torrent (Thermo Fisher, Inc.) and an Ion Proton sequencer (Thermo Fisher, Inc.). Then use

VILO^TMThe kit transcribes the RNA into cDNA. The cDNA was amplified using Ion Ampliseq Transcriptome Human Gene Expression Core panel, followed by ligation of an adaptor (adapter) and a barcode (barcode) to the amplicon and purification. The purified Library was quantified using an Ion Library Quantification kit (Thermo Fisher, Inc.). The library was then diluted to 100pM, pooled, and subjected to Ion Sphere using emulsion PCR^TMAmplified on particles (ISP) and enriched on IonChef (Thermo Fisher, Inc). The template positive ISP was then loaded onto an Ion PI chip and run on a Proton instrument (Proton instrument). AmpliSeq Comprehensive Cancer panel (409 tumor suppressor genes and oncogenes including FGFR3) or Ion AmpliSeq was used^TMCancer Hotspot Panel v2 performs targeted DNA sequencing of Cancer-associated genes.

RNA-Seq gene expression analysis was performed using the AmpliSeqRNA analysis plug-in the Torrent Suite software. The plug-in aligned the original sequence reads to a human reference genome containing 20,802 RefSeq transcripts (hg19 Ampliseq transgene _ ERCC _ v1.fasta) using the Torrent Mapping Alignment Program (TMAP). The number of reads per gene location is then counted to generate a raw count file and a normalized reads per million locations (RPM) per gene file. Tumor subtype assignment was performed using one of the nearest neighbor (oneNN) classifiers of MD anderson. Eight tumors showed Partial Remission (PR), four showed Stable Disease (SD), seven showed disease Progression (PD), and three were not classified (fig. 1).

To test for Differential Expression (DE) between before and after tumor treatment, a Bioconductor package DESeq with a Negative Binomial model (Negative Binomial model) was used. The Benjamini-Hochberg method was used to control the False Discovery Rate (FDR). Unsupervised cluster analysis of the baseline organization revealed the presence of two clusters: cluster 1 was enriched for responders (6 PR, 1 SD) compared to cluster 2(2 PR, 4 SD, 6 PD) (fig. 1). Genes significantly differentially expressed in

clusters

1 and 2 were extracted and analyzed by informaty Pathway Analysis (Sigma).

For pathway analysis, published gene expression signatures characteristic of immune infiltration, inflammatory cell trafficking, IFN pathway activation, TH1 pathway activation, and FGFR3 gene expression signatures were evaluated. Function and Pathway Analysis Using Ingeneity Path Analysis (IPA) software: (

Systems, CA) containing a database for identifying networks and pathways of interest in genomic data. The "transcription factor as a molecular species in the upstream regulator" classification within IPA was used to explain the biological properties of bladder tumor subtypes. For upstream modulator analysis, IPA performs statistical analysis on the overlap P value and activation Z score. Based on the IPA knowledge database, p-values and z-scores can be calculated based on how many targets (p-values) each transcription factor will wrap (over wrap) and the degree of consistency (z-score) of the known effects (activation or inhibition) of the targets in the gene list.

Gene signatures for identification of basal and luminal subtypes were used on baseline tissues to assign 22 paired tumors to molecular subtypes (fig. 2). Of the tumors that responded, 6 were lumens and 2 were bases (fig. 2); see also Denrauer 2014. Differential expression analysis of responders indicated that vofatamab affected inflammatory and immune pathways as well as immune cell trafficking (figure 3).

Taken together, these data indicate that luminal biology can sensitize subjects to combination therapy with vofatamab and pembrolizumab regardless of the presence of mutFGFR3, suggesting that this combination plays a role in wtFGFR3 mUC. The effect of this combination on tumors with basal biology also demonstrates effectiveness. These biological effects may also be important in early stage bladder cancer, e.g., non-muscle invasive bladder cancer (NMIBC). If vofatamab has an effect on immune cell trafficking, it may also show benefit after BCG treatment or in NMIBC in combination with BCG treatment.

Example 2: clinical use of vofatamab monotherapy and combination therapy with pembrolizumab in a subject with mUC Bed evaluation

FIERCE-22 phase 1b/2 clinical studies were designed to evaluate vofatamab in combination with pembrolizumab. Figure 4 shows a schematic of the study. Subjects received vofatamab monotherapy within a two-week lead (lead-in) and performed paired pre-and post-biopsy, followed by initiation of combination therapy. After the lead period, combination therapy was initiated.

A preliminary interim analysis was planned after the 26 patients were enrolled. 28 patients were enrolled in the study. A second interim analysis was planned after the 52 patients were enrolled. To qualify for inclusion, mUC patients need to be untreated with anti-PD-/L1, have measurable disease with an ECOG of greater than 2, progress on one or more lines of prior platinum-based chemotherapy or relapse within 12 months or less of (neo) adjuvant chemotherapy. The demographics and treatment history of the first 28 patients are shown in table 1.

Table 1: baseline demographics and treatment history：

The primary endpoints are safety and efficacy. Total response rate (ORR) was assessed by investigators using RECIST1.1, and any biopsy lesions were initially considered as non-targets for assessment. The treatment of the first 28 patients is shown in table 2. Median treatment exposure was prolonged and most patients began eight or more treatment cycles.

Table 2: treatment-period 2：

Anemia and exacerbation of tracheobronchitis

The clinical responses of the first 28 patients in phase 2 are shown in table 3, and the clinical responses of the evaluable population according to RECIST1.1 are shown in table 4.

Table 3: clinical response phase 2：

Table 4: clinical response-RECIST 1.1 evaluable population：

Consent was withdrawn from 1 patient before confirmation of PR and one patient had false progress on the next scan.

Patients in mut/fus have a short follow-up of about 4 months.

The most frequent Treatment Emergent Adverse Events (TEAE) in greater than or at least 15% of the 36 patients at stages 1b and 2 are shown in table 5. No cases of hyperphosphatemia or ocular or nail toxicity were reported. Most TEAEs were mainly grade 1, occurred early in the study, and were addressed in the study treatment.

Table 5: TEAE commonly occurring in > 15% of patients in 1b/2 phase：

Included 8 patients in stage 1 b.

1 of 4 occurred at stage 1 b.

The most frequently occurring vofatamab-associated TEAE in greater than or at least 15% of the 36 patients in stages 1b and 2 is shown in table 6. There are no vofatamab related TEAE reports of grade 3 or higher. The most common vofatamab-associated TEAEs are fatigue and diarrhea.

Table 6: vofatamab associated TEAE in > 15% of patients in 1b/2 phase：

Included 8 patients in stage 1 b.

The leading window of vofatamab was well tolerated and paired biopsies were safely obtained from most patients (80%). Combination therapy with pembrolizumab was equally well tolerated. No dose reduction of vofatamab occurred and there were few interruptions, 6 out of 303 cycles. Serious AEs occurring in > 2 patients were urinary tract infection (n ═ 2; 6%) and acute kidney injury (n ═ 2; 6%). All severe SEs were not associated with vofatamab.

In the RECIST1.1 evaluable phase 2 study population, ORR was approximately twice that of pembrolizumab alone. All 22 patients with evaluable lesions had an ORR of 40% (9 out of 22). The ORR of WT was 40% (6 out of 15), and that of Mut/Fus was 43% (3 out of 7). Thus, the response was similar regardless of FGFR3 Mut/Fus or WT status (fig. 5, 13). Patients treated with combination therapy showed a favorable trend for Progression Free Survival (PFS) relative to pembrolizumab alone (2.1 months; 95% CI 2.0,2.2) (5.9 months; 95% CI 3.6, 8.0). The DOR is shorter than the previously reported CPI alone and does not appear to predict OS. With at least 12 months of follow-up in all WT patients, median OS was not reached.

Lead vofatamab monotherapy induced up-regulation of genes associated with inflammatory responses and immune changes as well as immune pathway alterations in responders (figure 6). Clinical responses were associated with a subset of molecules, and elevated response rates (6 out of 8) were observed in a subset of lumens (i.e., in subjects with immunologically cold tumors) (fig. 7-12), suggesting that combination therapy may be particularly effective for this subtype. The presence of cancer-associated fibroblasts (CAF) appears to be a possible mechanism associated with a lack of response. Patients with p 53-like tumors showed poor response (figure 14).

Vofatamab inhibition by FGFR3 results in immunological changes. In a preliminary subtype study, vofatamab in combination with pembrolizumab improved the response in bladder cancer subtypes (except for the p 53-like group). Resistance to treatment may be associated with patients of the p 53-like subgroup. Several patients achieved sustained clinical benefit from treatment beyond RECIST1.1 progression.

Reference to the literature

1.American Cancer Society,Cancer Facts&Figures(2018)

2.Bai et al.Cancer Res 70(19):7630-7639(Aug.13,2010)

3.Bellmunt et al.J Clin Oncol 27(27):4454-4461(Aug.17,2009)

4.Brooks et al.Clin Cancer Res 18(7):1855-1862(Mar.2,2012)

5.Bumbaca et al.MAbs 3(4):376-386(July 1,2011)

6.Costantini et al.ScientificWorldJournal 11:1981-1994(Oct.26,2011)

7.Denrauer et al.Proc Natl Acad Sci USA 111(8):3110-3115(Feb.11,2014)

8.Eble et al.World Health Organization Classification of Tumors.Pathology and Genetics of Tumors of the Urinary System and Male Genital Organs.Lyon,France.IARC Press(2004)

9.Jemal et al.CA Cancer J Clin.61(2):69-90(Mar.-Apr.2011)

10.Lafitte et al.Mol.Cancer 12:83(July 31,2013)

11.Pardoll Nat Rev Cancer 12(4):252-264(Mar.22,2012)

12.Siefker-Radtke et al.J Clin Oncol 20(5):1361-1367(Mar.1,2002)

13.Sternberg et al.J Clin Oncol 19(10):2638-2646(May 15,2001)

14.Vallot et al.J Natl Cancer Inst 103(1):47-60(Dec.20,2010)

15.von der Maase et al.J Clin Oncol 18(17):3068-3077(Sept.2000)

16.Wu Nat Rev Cancer 5(9):713-725(Sept.2005)

17.Zhao et al.Clin Cancer Res 16(23):5750-5758(Dec.1,2010)

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Gly Arg Ile Tyr Pro Thr Ser Gly Ser Thr Asn Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Thr Tyr Gly Ile Tyr Asp Leu Tyr Val Asp Tyr Thr Glu Tyr

100 105 110

Val Met Asp Tyr Trp Gly Gln Gly Thr Leu Val

115 120

<210> 8

<211> 108

<212> PRT

<213> Artificial sequence

<220>

<223> light chain variable region

<400> 8

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Val Asp Thr Ser

20 25 30

Leu Ala Trp Tyr Lys Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Thr Gly His Pro Gln

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg

100 105

<210> 9

<211> 457

<212> PRT

<213> Artificial sequence

<220>

<223> heavy chain human IgG1

<400> 9

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Thr

20 25 30

Gly Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Arg Ile Tyr Pro Thr Ser Gly Ser Thr Asn Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Thr Tyr Gly Ile Tyr Asp Leu Tyr Val Asp Tyr Thr Glu Tyr

100 105 110

Val Met Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala

115 120 125

Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser

130 135 140

Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe

145 150 155 160

Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly

165 170 175

Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu

180 185 190

Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr

195 200 205

Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys

210 215 220

Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro

225 230 235 240

Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

245 250 255

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

260 265 270

Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

275 280 285

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

290 295 300

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

305 310 315 320

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

325 330 335

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

340 345 350

Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met

355 360 365

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

370 375 380

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

385 390 395 400

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu

405 410 415

Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

420 425 430

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

435 440 445

Lys Ser Leu Ser Leu Ser Pro Gly Lys

450 455

<210> 10

<211> 214

<212> PRT

<213> Artificial sequence

<220>

<223> light chain variable region + human Ig kappa chain C

<400> 10

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Val Asp Thr Ser

20 25 30

Leu Ala Trp Tyr Lys Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Thr Gly His Pro Gln

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

Claims

1. A method of treating luminal bladder cancer expressing wild-type FGFR3 in a subject in need thereof comprising administering to the subject a therapeutically effective amount of an FGFR3 inhibitor in combination with a therapeutically effective amount of a checkpoint inhibitor.

2. The method of claim 1, wherein the FGFR3 inhibitor is an antagonistic FGFR3 inhibitor.

3. The method of claim 2, wherein the antagonistic FGFR3 inhibitor is an antagonistic FGFR3 antibody.

4. The method of claim 3, wherein the antagonistic FGFR3 antibody comprises 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, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO. 3.

5. The method of claim 4, wherein the antagonistic FGFR3 antibody comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO 7.

6. The method of claim 3, wherein the antagonistic FGFR3 antibody comprises 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, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO. 6.

7. The method of claim 6, wherein the antagonistic FGFR3 antibody comprises a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO 8.

8. The method of claim 1, wherein the FGFR3 inhibitor is vofatamab.

9. The method of claim 1, wherein the checkpoint inhibitor is a PD1 inhibitor.

10. The method of claim 9, wherein the PD1 inhibitor is an antagonistic PD-L1 antibody.

11. The method of claim 10, wherein the antagonistic PD-L1 antibody is selected from the group consisting of MEDI-4736, RG7446, BMS-936559, MSB0010718C, and MPDL 3280A.

12. The method of claim 9, wherein the PD1 inhibitor is pembrolizumab (pembrolizumab).

13. A method of treating luminal bladder cancer expressing wild-type FGFR3 in a subject in need thereof comprising administering a therapeutically effective amount of an antagonistic FGFR3 inhibitor in combination with a therapeutically effective amount of a PD1 inhibitor.

14. The method of claim 13, wherein the antagonistic FGFR3 inhibitor is an antagonistic FGFR3 antibody.

15. The method of claim 14, wherein the antagonistic FGFR3 antibody comprises 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 a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO. 7.

16. The method of claim 14, wherein the antagonistic FGFR3 antibody comprises 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 a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO. 8.

17. The method of claim 13, wherein the FGFR3 inhibitor is vofatamab.

18. The method of claim 13, wherein the PD1 inhibitor is an antagonistic PD-L1 antibody.

19. The method of claim 18, wherein the antagonistic PD-L1 antibody is selected from the group consisting of MEDI-4736, RG7446, BMS-936559, MSB0010718C, and MPDL 3280A.

20. The method of claim 13, wherein the PD1 inhibitor is pembrolizumab.

21. A method of treating a subject in need thereof with a cancer that expresses wild-type FGFR3, the method comprising:

(a) screening the subject for a gene signature (signature) associated with fibroblasts associated with one or more cancers, or for p53 expression;

(b) determining whether the subject has the gene signature associated with the one or more cancer-associated fibroblasts or has p53 expression;

(c) determination based on step (b)

(i) Administering a therapeutically effective amount of an FGFR3 inhibitor in combination with a therapeutically effective amount of a checkpoint inhibitor if the subject does not have the gene signature or p53 expression, and

(ii) 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 if the subject has the gene signature or p53 expression.

22. The method of claim 21, wherein the FGFR3 inhibitor is an antagonistic FGFR3 antibody.

23. The method of claim 21, wherein the antagonistic FGFR3 antibody comprises 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 a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO. 7.

24. The method of claim 21, wherein the antagonistic FGFR3 antibody comprises 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 a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO. 8.

25. The method of claim 21, wherein the FGFR3 inhibitor is vofatamab.

26. The method of claim 21, wherein the checkpoint inhibitor is a PD1 inhibitor.

27. The method of claim 26, wherein the PD1 inhibitor is an antagonistic PD-L1 antibody selected from the group consisting of MEDI-4736, RG7446, BMS-936559, MSB0010718C, and MPDL 3280A.

28. The method of claim 21, wherein the PD1 inhibitor is pembrolizumab.

29. The method of claim 21, wherein the cancer is luminal bladder cancer.