Methods for treating muscle-invasive urothelial carcinoma or muscle-invasive bladder cancer with antibody drug conjugates (ADCs) that bind to 191P4D12 protein

Abstract

Provided herein are methods for treating urothelial or bladder cancer with antibody drug conjugates (ADCs) that bind to the 191P4D12 protein (Nectin-4).

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Application No. 63 / 296,831, filed January 5, 2022, and U.S. Application No. 63 / 304,136, filed January 28, 2022, the disclosures of each of which are incorporated herein by reference in their entireties.

[0002] Sequence Listing This application contains a computer-readable sequence listing that has been submitted herewith in XML file format, the entire contents of which are incorporated herein by reference. The Sequence Listing XML file submitted herewith is entitled "14369-279-228_SequenceListing.xml", was created on December 15, 2022, and is 34,925 bytes in size.

[0003] 1. Field Provided herein are methods for treating urothelial cancer or bladder cancer, such as muscle-invasive urothelial carcinoma (MIUC) or muscle-invasive bladder cancer (MIBC), with antibody drug conjugates (ADCs) that bind to the 191P4D12 protein (Nectin-4). [Background technology]

[0004] 2.Background 191P4D12 (also known as nectin-4) is a 66 kDa type I transmembrane protein that belongs to the nectin family of adhesion molecules. 191P4D12 is composed of an extracellular domain (ECD) containing three immunoglobulin (Ig)-like subdomains, a transmembrane helix, and an intracellular region (Takai et al., Annu Rev Cell Dev Biol (2008); 24:309-42). Nectins mediate Ca transport through both homophilic and heterophilic trans-interactions at adherens junctions, which can recruit cadherins and regulate cytoskeletal rearrangements. 2+It is thought to mediate cell-cell adhesion independent of the nuclei (Rikitake et al., Cell Mol Life Sci (2008); 65(2):253-63). The sequence identity of nectin-4 to other nectin family members is low, ranging from 25% to 30% in the ECD (Reymond et al., Biol Chem (2001); 276(46):43205-15).

[0005] The three Ig-like subdomains within the ECD of nectin-4 are designated V, C1, and C2. The C1 domain is involved in cis interactions (homodimerization), whereas the V domain of most nectin molecules contributes to trans interactions and cell-cell adhesion (Mandai et al., Curr Top Dev Biol (2015); 112:197-231; Takai et al., Nat Rev Mol Cell Biol (2008); 9(8):603-15).

[0006] Nectin-4 was originally identified by bioinformatics and cloned from human trachea (Reymond et al., J Biol Chem (2001) 276(46):43205-15). Nectin-4 was confirmed to be significantly upregulated in urothelial carcinoma using suppression subtractive hybridization in a pool of urothelial carcinoma specimens. Characterization of expression in multiple tumor samples, both at the ribonucleic acid (RNA) level and by immunohistochemistry (IHC), also showed high levels of nectin-4 in breast, pancreatic, lung, and other cancers (Challita-Eid et al., Cancer Res (2016);76(10):3003-13).

[0007] Nectin-4 has been shown to be expressed in several cancers, particularly urothelial, breast, lung, pancreatic, and ovarian cancers. Higher expression levels are associated with disease progression and / or poor prognosis (Fabre-Lafay et al., BMC Cancer (2007); 7:73).

[0008] urothelial cancer Urothelial carcinoma is the most common form of cancer occurring within the genitourinary tract and is a leading cause of morbidity and mortality. Approximately 151,000 new cases of bladder cancer, the most common primary site for urothelial carcinoma, are diagnosed annually in Europe (Ervik 2016). In the United States, the National Cancer Institute estimates that more than 79,000 new cases of bladder cancer were diagnosed in 2017, and more than 16,800 people died from the disease (National Cancer Institute 2018).

[0009] bladder cancer Bladder cancer accounts for approximately 5 percent of all new cancer cases in men (the fifth most common neoplasm) and 3 percent of all new cancer cases in women (the eighth most common neoplasm). Incidence rates are gradually increasing with the aging population. The American Cancer Society (cancer.org) estimates that there are 81,400 new cases annually, including 62,100 in men and 19,300 in women, accounting for 4.5% of all cancer cases. The age-adjusted incidence rate in the United States is 20 per 100,000 for both men and women. Bladder cancer is estimated to cause 17,980 deaths annually (13,050 in men and 4,930 in women), accounting for 3% of all cancer-related deaths. Bladder cancer incidence and mortality rates increase significantly with age, making it a growing problem with an aging population. Approximately 580,000 people will be diagnosed with bladder cancer worldwide in 2020, and bladder cancer is estimated to cause approximately 210,000 deaths worldwide.

[0010] Most bladder cancers recur in the bladder. Bladder cancer is managed with transurethral cystectomy (TUR) combined with intravesical chemotherapy or intravesical immunotherapy. Multifocal and recurrent bladder cancers demonstrate the limitations of TUR. Most muscle-invasive cancers are not cured by TUR alone. Radical cystectomy and urinary diversion are the most effective means of eliminating cancer, but inevitably affect urinary and sexual function. There remains a significant need for treatments that benefit bladder cancer patients.

[0011] There is a significant need for additional treatment methods for urothelial and bladder cancer, including the use of antibodies and antibody-drug conjugates as therapeutics.

[0012] Muscle-invasive bladder cancer (MIBC) and muscle-invasive urothelial carcinoma (MIUC) Muscle-invasive urothelial carcinoma (MIUC) includes cancer originating in any part of the urinary tract: the renal pelvis or upper tract, the bladder, and the lower tract (urethra). Muscle-invasive bladder cancer (MIBC) refers to cancer of the bladder, excluding the upper and lower tracts. Up to 25% of all patients diagnosed with urothelial carcinoma will present with muscle-invasive disease, which carries a significant risk of progression or metastasis. MIBC has a 5-year mortality rate of approximately 69% (American Cancer Society (ACS), Survival rates for bladder cancer, 2019, www.cancer.org / cancer / bladder-cancer / detection-diagnosis-staging / survival-rates.html. Accessed: February 23, 2018). Prognosis and recurrence vary depending on the stage of disease and other prognostic features, including lymph node involvement, lymphovascular invasion, tumor stage, presence of variant histology, and molecular subtype (Lotan et al., J. Clin. Oncol., 2005, 23(27):6533-9; Karakiewicz et al., J Urol., 2006, 176(4 Pt 1):1354-61; Choi 2014).

[0013] The treatment of MIBC is complex and requires multidisciplinary collaboration among surgeons, radiation therapists, and medical oncologists (Aragon-Ching et al., Am Soc Clin Oncol Educ Book, 2018, 38:307-18). Cisplatin-based neoadjuvant chemotherapy (NAC) followed by radical cystectomy (RC) and lymphadenectomy has been the standard of care in MIBC for nearly 20 years, as established by evidence from several randomized trials and a large meta-analysis first published by the Advanced Bladder Cancer Meta-Analysis Collaboration in 2003 and updated in 2005 (Advanced Bladder Cancer Meta-analysis, Eur Urol., 2005, 48(2):202-5). The final version of the meta-analysis included 11 randomized trials and 3,005 patients and compared platinum-based NAC (all but one trial involved cisplatin) plus definitive local treatment (cystectomy or radiation) with the same local treatment alone. Results showed a significant benefit for patients receiving cisplatin-based combination NAC, with a 14% reduction in risk of death (HR, 0.86; 95% confidence interval [CI], 0.77–0.95; P = 0.003), which resulted in a 5% improvement in OS and a 9% improvement in disease-free survival (DFS) at 5 years. For patients receiving cisplatin-based NAC, 5-year OS was 50% compared with 45% for patients receiving definitive local therapy alone. No survival benefit was demonstrated for patients receiving carboplatin-based combinations. More recently, additional NAC regimens, including accelerated or dose-dense MVAC and combinations of gemcitabine and cisplatin (GC), have been studied with the aim of improving patient tolerability and shortening treatment duration with comparable pathological response rates. Approximately 30% to 40% of patients achieved a pCR rate, and 50% had pathological downstaging with accelerated or dose-dense MVAC (Choueiri et al., J Clin Oncol, 2014, 32(18):1889-94; Plimack et al., J Clin Oncol., 2014, 32(18):1895-901).

[0014] Although NAC before RC is effective, residual high-risk disease (≥pT2) still exists in >50% of patients and is associated with a poor prognosis. Additionally, many patients are ineligible for surgery or cisplatin, and consideration of bladder-preserving strategies is not only increasingly recognized as an optimal treatment option, but should also feature prominently in the range of management options presented to patients at diagnosis (Aragon-Ching et al., Am Soc Clin Oncol Educ Book, 2018, 38:307-18). Nevertheless, these trials are limited to biomarker-selected patients predicted to respond adequately to neoadjuvant treatment.

[0015] Apart from chemotherapy, immunotherapy has been successfully used in locally advanced and metastatic bladder cancer and is moving into the MIBC space as monotherapy or in combination with chemotherapy. Two recent investigator-initiated trials were designed to obtain biomarker results and evaluate the efficacy of single-agent programmed cell death 1 / programmed cell death-ligand 1 (PD1 / PD-L1) inhibitors in patients with MIBC in the neoadjuvant setting. Both have published preliminary data. In the PURE-01 trial, patients with T2 T3bN0M0 disease, regardless of cisplatin eligibility, received three cycles of 200 mg pembrolizumab before RC (Necchi et al., J Clin Oncol., 2018, 36(34):3353-60). Pembrolizumab was administered safely and resulted in a 42% pCR rate among 50 treated patients, with a 54.3% pCR rate and a 65.7% pathological downstaging rate in patients with PD-L1-positive disease. Immune-mediated adverse events (imAEs) were reported but did not delay planned surgery. Postoperative complications were similar to those reported in the literature and were associated with either open or robotic surgery. Neoadjuvant pembrolizumab did not impair intraoperative performance measures, such as the number of lymph nodes removed, surgical margins, and operative time. In the ABACUS trial, 69 cisplatin-ineligible patients with T2 T4N0M0 MIBC were treated with two cycles of 1200 mg neoadjuvant atezolizumab before RC, with no significant safety findings (Powles et al., 2018, J Clin Oncol., 36(Suppl 15):Abstract 4506).

[0016] pCR was observed in 29% of patients and 40% of patients with PD-L1-positive disease, compared with only 16% in the PD-L1-negative cohort. A total of 39% of patients downgraded to non-muscle-invasive disease. Both trials lack the long-term follow-up data necessary to assess PFS and OS in the postoperative setting for patients receiving neoadjuvant immunotherapy.

[0017] Currently, there are no standard neoadjuvant or perioperative treatment options for patients with MIBC who are cisplatin-ineligible. Therefore, newer treatment approaches, including novel drug combination strategies, are needed for this patient population. Summary of the Invention

[0018] 3. Overview Provided herein are methods for treating various cancers in human subjects, such as urothelial cancer and bladder cancer, including subjects with muscle-invasive bladder cancer (MIBC) and muscle-invasive urothelial carcinoma (MIUC) that are cisplatin-ineligible, using antibody drug conjugates (ADCs) that bind to 191P4D12.

[0019] Embodiment 1. A method of treating muscle-invasive urothelial carcinoma (MIUC) or muscle-invasive bladder cancer (MIBC) in a human subject, comprising: (a) administering to the subject an effective amount of an antibody-drug conjugate (ADC), wherein the ADC comprises an antibody or antigen-binding fragment thereof that binds to 191P4D12 (Nectin-4) conjugated to one or more units of monomethyl auristatin E (MMAE), wherein the subject is ineligible to receive cisplatin treatment (cisplatin-ineligible), and wherein the pathological complete response rate (pCRR) is at least 30%.

[0020] Embodiment 2. A method of treating muscle-invasive urothelial carcinoma (MIUC) or muscle-invasive bladder cancer (MIBC) in a human subject, comprising administering to the subject an effective amount of an antibody-drug conjugate (ADC), wherein the ADC comprises an antibody or antigen-binding fragment thereof that binds to 191P4D12 (Nectin-4) conjugated to one or more units of monomethyl auristatin E (MMAE), wherein the subject is ineligible to receive cisplatin treatment (cisplatin-ineligible), and wherein the pathological downstaging rate (pDSR) is at least 50%.

[0021] Embodiment 3. The method of embodiment 1 or embodiment 2, wherein the method comprises administering three cycles of an ADC to the subject.

[0022] Embodiment 4. The method of any one of embodiments 1-3, wherein the treatment of cancer further comprises radical cystectomy and pelvic lymph node dissection (RC+PLND).

[0023] Embodiment 5. The method of embodiment 4, wherein the human subject undergoes RC+PLND about 4 to about 12 weeks after the ADC is administered to the human subject.

[0024] Embodiment 6. (b) The method of embodiment 1 or 2, further comprising performing a radical cystectomy and pelvic lymph node dissection (RC+PLND) on the subject.

[0025] Embodiment 7. The method of embodiment 6, wherein (b) is performed about 4 to about 12 weeks after (a).

[0026] Embodiment 8. A method of treating muscle-invasive urothelial carcinoma (MIUC) or muscle-invasive bladder cancer (MIBC) in a human subject, comprising administering to the subject multiple cycles of an effective amount of an antibody drug conjugate (ADC); (a) the ADC comprises an antibody or antigen-binding fragment thereof that binds to 191P4D12 (Nectin-4) conjugated to one or more units of monomethyl auristatin E (MMAE); (b) the subject is ineligible to receive cisplatin treatment (cisplatin-ineligible); (c) The method, wherein the number of cycles of ADC treatment administered to the cisplatin-ineligible subject is equal to or less than the number of cycles of standard of care (SOC) therapy used to treat cisplatin-eligible subjects with MIUC or MIBC.

[0027] Embodiment 9 The method of embodiment 8, wherein the SOC therapy for the cisplatin-eligible subject comprises cisplatin.

[0028] Embodiment 10. The method of embodiment 8, wherein the SOC therapy for the cisplatin-eligible subject comprises (i) methotrexate, vinblastine, doxorubicin, and cisplatin (MVAC), or (ii) gemcitabine and cisplatin.

[0029] Embodiment 11. The method of embodiment 8, wherein the SOC therapy for the cisplatin-eligible subject comprises administering a programmed cell death 1 (PD-1) or programmed cell death-ligand 1 (PD-L1) inhibitor.

[0030] Embodiment 12. The method of any one of embodiments 8 to 11, wherein the number of cycles of ADC treatment administered to the cisplatin-ineligible subject is 3 or 4.

[0031] Embodiment 13 The method of any one of embodiments 8 to 11, wherein the number of cycles of ADC treatment administered to the cisplatin-ineligible subject is 3.

[0032] Embodiment 14. A method of administering neoadjuvant or perioperative therapy for treating muscle-invasive urothelial carcinoma (MIUC) or muscle-invasive bladder cancer (MIBC) in a human subject, comprising administering to the subject multiple cycles of an effective amount of an antibody drug conjugate (ADC); (a) the ADC comprises an antibody or antigen-binding fragment thereof that binds to 191P4D12 (Nectin-4) conjugated to one or more units of monomethyl auristatin E (MMAE); (b) the subject is ineligible to receive cisplatin treatment (cisplatin-ineligible); (c) A method in which the subject remains eligible for radical cystectomy and pelvic lymph node dissection (RC+PLND) surgery after multiple cycles of treatment with the ADC.

[0033] Embodiment 15 The method of embodiment 14, wherein the ADC is administered as neoadjuvant therapy prior to surgery.

[0034] Embodiment 16 The method of embodiment 14, wherein the ADC is administered before and after surgery.

[0035] Embodiment 17. A method of treating muscle-invasive urothelial carcinoma (MIUC) or muscle-invasive bladder cancer (MIBC) in a human subject, comprising administering to the subject multiple cycles of an effective amount of an antibody drug conjugate (ADC); (a) the ADC comprises an antibody or antigen-binding fragment thereof that binds to 191P4D12 (Nectin-4) conjugated to one or more units of monomethyl auristatin E (MMAE); (b) the subject is ineligible to receive cisplatin treatment (cisplatin-ineligible); (c) the efficacy of treatment of the subject with the effective amount of the ADC is similar to the efficacy of treatment observed in cisplatin-eligible patients treated with a standard of care (SOC) regimen used to treat cisplatin-eligible subjects with MIUC or MIBC.

[0036] Embodiment 18. The method of embodiment 17, wherein the efficacy of the treatment is at least as effective as the efficacy of the treatment observed in the SOC for cisplatin-eligible subjects.

[0037] Embodiment 19. The method of embodiment 17 or 18, wherein the measure of efficacy of the treatment is one or more of the following: pathological complete response rate (pCRR), pathological downstaging rate (pDSR), disease-free survival (DFS), recurrence-free survival (EFS), overall survival (OS), progression-free survival (PFS), and duration of response (DoR).

[0038] Embodiment 20 The method of embodiment 19, wherein the pCRR for cisplatin-ineligible subjects is at least 30%.

[0039] Embodiment 21 The method of embodiment 19, wherein the pDSR for cisplatin-ineligible subjects is at least 50%.

[0040] Embodiment 22 The method of any one of embodiments 18-21, wherein the SOC therapy for a cisplatin-eligible subject comprises cisplatin.

[0041] Embodiment 23. The method of embodiment 22, wherein the SOC therapy for the cisplatin-eligible subject comprises (i) methotrexate, vinblastine, doxorubicin, and cisplatin (MVAC), or (ii) gemcitabine and cisplatin.

[0042] Embodiment 24. The method of embodiment 18, wherein the SOC therapy for the cisplatin-eligible subject comprises a programmed cell death 1 (PD-1) or programmed cell death-ligand 1 (PD-L1) inhibitor.

[0043] Embodiment 25. The method of embodiment 24, wherein the PD-1 inhibitor is nivolumab or pembrolizumab.

[0044] Embodiment 26. The method of embodiment 24, wherein the PD-L1 inhibitor is selected from the group consisting of atezolizumab, avelumab, and durvalumab.

[0045] Embodiment 27. The method of any one of embodiments 1 to 26, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising a CDR comprising the amino acid sequence of the complementarity-determining region (CDR) of the heavy chain variable region set forth in SEQ ID NO: 22, and a light chain variable region comprising a CDR comprising the amino acid sequence of the CDR of the light chain variable region set forth in SEQ ID NO: 23.

[0046] Embodiment 28. The antibody or antigen-binding fragment thereof comprises CDR-H1 comprising the amino acid sequence of SEQ ID NO:9, CDR-H2 comprising the amino acid sequence of SEQ ID NO:10, CDR-H3 comprising the amino acid sequence of SEQ ID NO:11, CDR-L1 comprising the amino acid sequence of SEQ ID NO:12, CDR-L2 comprising the amino acid sequence of SEQ ID NO:13, and CDR-L3 comprising the amino acid sequence of SEQ ID NO:14; or The method of any one of embodiments 1 to 27, wherein the antibody or antigen-binding fragment thereof comprises CDR-H1 having the amino acid sequence of SEQ ID NO: 16, CDR-H2 having the amino acid sequence of SEQ ID NO: 17, CDR-H3 having the amino acid sequence of SEQ ID NO: 18, CDR-L1 having the amino acid sequence of SEQ ID NO: 19, CDR-L2 having the amino acid sequence of SEQ ID NO: 20, and CDR-L3 having the amino acid sequence of SEQ ID NO: 21.

[0047] Embodiment 29. The antibody or antigen-binding fragment thereof comprises CDR-H1 consisting of the amino acid sequence of SEQ ID NO:9, CDR-H2 consisting of the amino acid sequence of SEQ ID NO:10, CDR-H3 consisting of the amino acid sequence of SEQ ID NO:11, CDR-L1 consisting of the amino acid sequence of SEQ ID NO:12, CDR-L2 consisting of the amino acid sequence of SEQ ID NO:13, and CDR-L3 consisting of the amino acid sequence of SEQ ID NO:14; or The method of any one of embodiments 1 to 27, wherein the antibody or antigen-binding fragment thereof comprises CDR-H1 consisting of the amino acid sequence of SEQ ID NO: 16, CDR-H2 consisting of the amino acid sequence of SEQ ID NO: 17, CDR-H3 consisting of the amino acid sequence of SEQ ID NO: 18, CDR-L1 consisting of the amino acid sequence of SEQ ID NO: 19, CDR-L2 consisting of the amino acid sequence of SEQ ID NO: 20, and CDR-L3 consisting of the amino acid sequence of SEQ ID NO: 21.

[0048] Embodiment 30. The method of any one of embodiments 1 to 29, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 22 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 23.

[0049] Embodiment 31. The method of any one of embodiments 1 to 30, wherein the antibody comprises a heavy chain comprising the amino acid sequence ranging from amino acid 20 (glutamic acid) to amino acid 466 (lysine) of SEQ ID NO:7, and a light chain comprising the amino acid sequence ranging from amino acid 23 (aspartic acid) to amino acid 236 (cysteine) of SEQ ID NO:8.

[0050] Embodiment 32. The method of any one of embodiments 1 to 30, wherein the antigen-binding fragment is a Fab, F(ab')2, Fv, or scFv.

[0051] Embodiment 33. The method of any one of embodiments 1 to 31, wherein the antibody is a fully human antibody.

[0052] Embodiment 34. The method of any one of embodiments 1 to 31 and 33, wherein the antibody is an IgG1 and the light chain is a kappa light chain.

[0053] Embodiment 35. The method of any one of embodiments 1 to 34, wherein the antibody or antigen-binding fragment thereof is recombinantly produced.

[0054] Embodiment 36 The method of any one of embodiments 1 to 35, wherein the antibody or antigen-binding fragment is conjugated to each unit of MMAE via a linker.

[0055] Embodiment 37. The method of embodiment 36, wherein the linker is an enzyme-cleavable linker, and the linker forms a bond with a sulfur atom of the antibody or antigen-binding fragment thereof.

[0056] Embodiment 38. The method of embodiment 36 or 37, wherein the linker has the formula -Aa-Ww-Yy-, where -A- is a Stretcher unit, a is 0 or 1, -W- is an amino acid unit, w is an integer ranging from 0 to 12, -Y- is a Spacer unit, and y is 0, 1, or 2.

[0057] Embodiment 39. The Stretcher unit has the structure of formula (1) below, the amino acid unit is valine-citrulline, and the Spacer unit is a PAB group having the structure of formula (2) below: TIFF2025501978000002.tif74128 The method described in embodiment 38.

[0058] Embodiment 40. The method of embodiment 38 or 39, wherein the Stretcher unit forms a bond with a sulfur atom of the antibody or antigen-binding fragment thereof, and the Spacer unit is linked to MMAE via a carbamate group.

[0059] Embodiment 41. The method of any one of embodiments 1 to 40, wherein the ADC comprises 1 to 20 units of MMAE per antibody or antigen-binding fragment thereof.

[0060] Embodiment 42. The method of any one of embodiments 1 to 41, wherein the ADC comprises 1 to 10 units of MMAE per antibody or antigen-binding fragment thereof.

[0061] Embodiment 43. The method of any one of embodiments 1 to 42, wherein the ADC comprises 2 to 8 units of MMAE per antibody or antigen-binding fragment thereof.

[0062] Embodiment 44. The method of any one of embodiments 1 to 43, wherein the ADC comprises 3 to 5 units of MMAE per antibody or antigen-binding fragment thereof.

[0063] Embodiment 45. The ADC has the structure: TIFF2025501978000003.tif36160, 43. The method of any one of embodiments 1 to 42, wherein L- represents an antibody or antigen-binding fragment thereof, and p is 1 to 10.

[0064] Embodiment 46. The method of embodiment 45, wherein p is 2 to 8.

[0065] Embodiment 47. The method of embodiment 45 or 46, wherein p is 3 to 5.

[0066] Embodiment 48. The method of any one of embodiments 45 to 47, wherein p is 3 to 4.

[0067] Embodiment 49. The method of any one of embodiments 45 to 48, wherein p is about 4.

[0068] Embodiment 50. The method of any one of embodiments 45 to 48, wherein the effective dose of the antibody drug conjugate has a mean p value of about 3.8.

[0069] Embodiment 51. The method of any one of embodiments 1 to 50, wherein the ADC is formulated in a pharmaceutical composition comprising L-histidine, polysorbate-20 (TWEEN-20), and trehalose dehydrate.

[0070] Embodiment 52. The method of any one of embodiments 1-51, wherein the ADC is formulated in a pharmaceutical composition comprising about 20 mM L-histidine, about 0.02% (w / v) TWEEN-20, about 5.5% (w / v) trehalose dihydrate, and hydrochloride, and the pH of the pharmaceutical composition is about 6.0 at 25°C.

[0071] Embodiment 53. The method of any one of embodiments 1-51, wherein the ADC is formulated in a pharmaceutical composition comprising about 9 mM histidine, about 11 mM histidine hydrochloride monohydrate, about 0.02% (w / v) TWEEN-20, and about 5.5% (w / v) trehalose dihydrate, and wherein the pH of the pharmaceutical composition is about 6.0 at 25°C.

[0072] Embodiment 54. The ADC has the structure: TIFF2025501978000004.tif36160, 54. The method of any one of embodiments 1-53, wherein L- represents an antibody or antigen-binding fragment thereof; p is about 3 to about 4; the antibody comprises a heavy chain comprising an amino acid sequence ranging from amino acid 20 (glutamic acid) to amino acid 466 (lysine) of SEQ ID NO: 7 and a light chain comprising an amino acid sequence ranging from amino acid 23 (aspartic acid) to amino acid 236 (cysteine) of SEQ ID NO: 8; and the ADC is administered at a dose of about 1.25 mg / kg of subject body weight, wherein the dose is administered by IV infusion on days 1 and 8 of a 3-week cycle.

[0073] Embodiment 55. The method of any one of embodiments 1 to 54, wherein the subject has cT2-T4aN0M0 stage MIBC.

[0074] Embodiment 56. The method of any one of embodiments 1-55, wherein a subject is considered cisplatin-ineligible if they meet one or more of the following criteria: (a) a GFR of less than 60 mL / min but greater than or equal to 30 mL / min, where GFR is measured by the Cockcroft-Gault formula, the Modification of Diet in Renal Disease formula (MDRD), or a 24-hour urine collection; (b) an ECOG performance status of 2; (c) hearing loss of grade 2 or greater per NCI CTCAE version 4.03; and (d) NYHA class III heart failure.

[0075] Embodiment 57. The method of any one of embodiments 1 to 56, wherein the subject has not received prior systemic treatment for MIBC, chemoradiotherapy, and / or radiation therapy.

[0076] Embodiment 58. The method of any one of embodiments 1 to 57, wherein the subject is not receiving an immune checkpoint inhibitor (CPI).

[0077] Embodiment 59. The method of embodiment 58, wherein the CPI is a programmed cell death 1 (PD-1) inhibitor or a programmed cell death-ligand 1 (PD-L1) inhibitor.

[0078] Embodiment 60. The method of any one of embodiments 1 to 59, wherein the subject has not received a CD137 agonist, a CTLA-4 inhibitor, or an OX-40 agonist.

[0079] Embodiment 61. The method of any one of embodiments 1 to 60, wherein the effective amount of the ADC is about 1 to about 10 mg / kg of the subject's body weight, about 1 to about 5 mg / kg of the subject's body weight, about 1 to about 2.5 mg / kg of the subject's body weight, or about 1 to about 1.25 mg / kg of the subject's body weight.

[0080] Embodiment 62. The method of any one of embodiments 1-60, wherein the effective amount of the ADC is about 0.25 mg / kg, about 0.5 mg / kg, about 0.75 mg / kg, about 1.0 mg / kg, about 1.25 mg / kg, about 1.5 mg / kg, about 1.75 mg / kg, about 2.0 mg / kg, about 2.25 mg / kg, or about 2.5 mg / kg of the subject's body weight.

[0081] Embodiment 63 The method of any one of embodiments 1 to 60, wherein the effective amount of the ADC is about 1 mg / kg of subject body weight.

[0082] Embodiment 64. The method of any one of embodiments 1 to 60, wherein the effective amount of the ADC is about 1.25 mg / kg of subject body weight.

[0083] Embodiment 65. (a) The method of any one of embodiments 1 to 60, wherein an effective amount of the ADC is administered to the subject on days 1 and 8 of a three-week cycle for a total of three cycles.

[0084] Embodiment 66. The method of embodiment 65, wherein the effective amount of the ADC is about 1.25 mg / kg of the subject's body weight.

[0085] Embodiment 67. (a) The method of embodiment 4 or 5, wherein an effective amount of an ADC is administered to the subject on days 1 and 8 of a 3-week cycle for a total of 3 cycles.

[0086] Embodiment 68. The method of any one of embodiments 4, 5, or 67, wherein the method further comprises (b) administering to the subject an effective amount of an ADC on days 1 and 8 of every 3-week cycle, approximately 8 weeks after the subject undergoes RC+PLND, for a total of 6 cycles.

[0087] Embodiment 69. The method of embodiment 67 or 68, wherein the effective amount of the ADC is about 1.25 mg / kg of the subject's body weight.

[0088] Embodiment 70. (a) The method of embodiment 6 or 7, wherein an effective amount of the ADC is administered to the subject on days 1 and 8 of a 3-week cycle for a total of 3 cycles.

[0089] Embodiment 71. The method of any one of embodiments 5, 7, or 70, further comprising administering to the subject about 8 weeks after (c)(b) an effective amount of an ADC on days 1 and 8 of every 3-week cycle, for a total of 3 cycles.

[0090] Embodiment 72. The method of embodiment 70 or 71, wherein the effective amount of ADC is about 1.25 mg / kg of subject body weight.

[0091] Embodiment 73. The method of any one of embodiments 8 to 64, wherein an effective amount of the ADC is administered to the human subject on days 1 and 8 of a 3-week cycle for a total of 3 cycles.

[0092] Embodiment 74. The method of any one of embodiments 1 to 73, wherein the ADC is administered by intravenous (IV) injection or infusion.

[0093] Embodiment 75. The method of any one of embodiments 1 to 74, wherein the cancer is muscle-invasive urothelial carcinoma (MIUC).

[0094] Embodiment 76. The method of any one of embodiments 1 to 74, wherein the cancer is muscle-invasive bladder cancer (MIBC).

[0095] Embodiment 77. The method of any one of embodiments 1 to 76, wherein the ADC is enfortumab vedotin (EV).

[0096] Embodiment 78. The method of any one of embodiments 1 to 77, wherein the overall survival of the subject is extended by at least 2 months, at least 4 months, at least 6 months, at least 8 months, at least 10 months, or at least 12 months.

[0097] Embodiment 79. The method of any one of embodiments 1 to 78, wherein the ADC is administered as monotherapy. [Brief explanation of the drawings]

[0098] 4. Brief description of the drawings [Figure 1A-1] Figures 1A-1E depict the nucleotide and amino acid sequences of the Nectin-4 protein (Figure 1A), the nucleotide and amino acid sequences of the heavy chain (Figure 1B) and light chain (Figure 1C) of Ha22-2(2.4)6.1, and the amino acid sequences of the heavy chain (Figure 1D) and light chain (Figure 1E) of Ha22-2(2.4)6.1. [Figure 1A-2] See legend to Figure 1A-1. [Figure 1B] See legend to Figure 1A-1. [Figure 1C] See legend to Figure 1A-1. [Figure 1D] See legend to Figure 1A-1. [Figure 1E] See legend to Figure 1A-1. [Figure 2] Draw the overall study design for the clinical trial described in Section 6.1. TURBT, transurethral resection of bladder tumor. [Figure 3-1] Figure 3 depicts the European Organization for Research and Treatment of Cancer (EORTC) Core Quality of Life (QLQ-C-30) assessment (EORTC-QLQ-C-30, current version, version 3), as described in Section 6.1. [Figure 3-2] See description of Figure 3-1. [Figure 4-1] Figure 4 depicts the EuroQol-5 items (EQ-5D-5L) described in Section 6.1. [Figure 4-2] See description of Figure 4-1. [Figure 5]Delineate MIUC disease stages (shown in Figure 5 are Tis, Ta, T1, T2a, T2b, T3a, T3b, T4a, and T4b) relative to the anatomy of the bladder and pelvic region as described in Section 6.2 (see, e.g., www.cancer.org / cancer / bladder-cancer / detection-diagnosis-staging / staging.html). [Figure 6] Plot the recurrence-free survival (EFS) observed in the clinical trials described in Section 6.2. MIBC EV Mono, muscle-invasive bladder cancer treated with enfortumab vedotin monotherapy. [Figure 7] Draw the overall study design for the clinical trial described in Section 6.3. TURBT, transurethral resection of bladder tumor. DETAILED DESCRIPTION OF THE INVENTION

[0099] 5. Detailed Description Before further describing the present disclosure, it is to be understood that the disclosure is not limited to the particular embodiments described herein, and that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

[0100] 5.1 Definition The techniques and procedures described or referenced herein include those that are generally well understood and / or commonly employed by those of skill in the art using conventional methodology, such as, for example, the widely utilized methodologies described in Sambrook et al., Molecular Cloning: A Laboratory Manual (3rd ed. 2001), Current Protocols in Molecular Biology (Ausubel et al. eds., 2003), Therapeutic Monoclonal Antibodies: From Bench to Clinic (An ed. 2009), Monoclonal Antibodies: Methods and Protocols (Albitar ed. 2010), and Antibody Engineering Vols 1 and 2 (Kontermann and Dubel eds., 2nd ed. 2010).

[0101] Unless otherwise defined herein, technical and scientific terms used in this description have the meanings that are commonly understood by those skilled in the art. For the purposes of interpreting this specification, the following explanations of terms shall apply, and whenever appropriate, terms used in the singular shall also include the plural and vice versa. In the event that the explanations of terms set forth conflict with documents incorporated herein by reference, the explanations of terms set forth below shall prevail.

[0102] The terms "antibody," "immunoglobulin," or "Ig" are used interchangeably herein and are used in the broadest sense, specifically encompassing, for example, monoclonal antibodies (including agonist, antagonist, neutralizing, full-length, or intact monoclonal antibodies), antibody compositions with polyepitopic or monoepitopic specificity, polyclonal or univalent antibodies, multivalent antibodies, multispecific antibodies formed from at least two intact antibodies (e.g., bispecific antibodies, so long as they exhibit the desired biological activity), single-chain antibodies, and fragments thereof, as described below. Antibodies may be human, humanized, chimeric, and / or affinity-matured antibodies, as well as antibodies from other species, e.g., mouse and rabbit. The term "antibody" is intended to include polypeptide products of B cells within the immunoglobulin class of polypeptides that can bind to a specific molecular antigen and are composed of two identical pairs of polypeptide chains, each pair having one heavy chain (about 50-70 kDa) and one light chain (about 25 kDa), with the amino-terminal portion of each chain containing a variable region of about 100 to about 130 or more amino acids and the carboxy-terminal portion of each chain containing a constant region. See, e.g., Antibody Engineering (Borrebaeck ed., 2nd ed. 1995) and Kuby, Immunology (3rd ed. 1997). In specific embodiments, specific molecular antigens can be bound by the antibodies provided herein, including polypeptides or epitopes. Antibodies also include, but are not limited to, synthetic antibodies, recombinantly produced antibodies, camelized antibodies, intrabodies, anti-idiotypic (anti-Id) antibodies, and functional fragments (e.g., antigen-binding fragments) of any of the above, where a functional fragment refers to a portion of an antibody heavy or light chain polypeptide that retains some or all of the binding activity of the antibody from which the fragment is derived. Non-limiting examples of functional fragments (e.g., antigen-binding fragments) include single-chain Fvs (scFvs) (including, e.g., monospecific, bispecific, etc.), Fab fragments, F(ab') fragments, F(ab') fragments, F(ab') fragments, disulfide-linked Fvs (dsFvs), Fd fragments, Fv fragments, diabodies, triabodies, tetrabodies, and minibodies.In particular, antibodies provided herein include immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, such as molecules containing an antigen-binding domain or site that binds to an antigen (e.g., one or more CDRs of an antibody). Such antibody fragments can be found, for example, in Harlow and Lane, Antibodies: A Laboratory Manual (1989), Mol. Biology and Biotechnology: A Comprehensive Desk Reference (Myers ed., 1995), Huston et al., 1993, Cell Biophysics 22:189-224, Pluckthun and Skerra, 1989, Meth. Enzymol. 178:497-515, and Day, Advanced Immunochemistry (2d ed. 1990). The antibodies provided herein can be of any class (e.g., IgG, IgE, IgM, IgD, and IgA) or any subclass (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2) of immunoglobulin molecule. The antibody can be an agonist or antagonist antibody.

[0103] The term "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible minor naturally occurring mutations. Monoclonal antibodies are highly specific, being directed against a single antigenic site. In contrast to polyclonal antibody preparations, which may include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen.

[0104] An "antigen" is a structure to which an antibody can selectively bind. Target antigens can be polypeptides, carbohydrates, nucleic acids, lipids, haptens, or other natural or synthetic compounds. In some embodiments, the target antigen is a polypeptide. In certain embodiments, the antigen is associated with a cell, e.g., present on or within a cell, e.g., on or within a cancer cell.

[0105] An "intact" antibody is one that comprises an antigen-binding site as well as a CL and at least heavy chain constant regions, CH1, CH2, and CH3. The constant region may comprise a human constant region or an amino acid sequence variant thereof. In certain embodiments, an intact antibody has one or more effector functions.

[0106] The terms "antigen-binding fragment," "antigen-binding domain," "antigen-binding region," and similar terms refer to a portion of an antibody (e.g., a CDR) that contains amino acid residues that interact with an antigen and confer specificity and affinity for the antigen to the binding agent. As used herein, "antigen-binding fragment" includes a portion of an intact antibody, e.g., an "antibody fragment" that contains the antigen-binding or variable region of the intact antibody. Examples of antibody fragments include Fab, Fab', F(ab')2, and Fv fragments, diabodies, and di-diabodies (e.g., Holliger et al., 1993, Proc. Natl. Acad. Sci. 90:6444-48; Lu et al., 2005, J. Biol. Chem. 280:19665-72; Hudson et al., 2005, J. Biol. Chem. 280:19665-72). al., 2003, Nat. Med. 9:129-34, WO 93 / 11161, and U.S. Pat. Nos. 5,837,242 and 6,492,123), single chain antibody molecules (see, e.g., U.S. Pat. Nos. 4,946,778, 5,260,203, 5,482,858, and 5,476,786), dual variable domain antibodies (see, e.g., U.S. Pat. No. 7,612,181), single variable domain antibodies (sdAbs) (see, e.g., Woolven et al., 1999, Immunogenetics 50:98-101, and Streltsov et al., 2004, Proc Natl Acad Sci USA.101:12444-49), as well as multispecific antibodies formed from antibody fragments.

[0107] The terms "bind" or "binding" refer to interactions between molecules, including, for example, forming a complex. The interaction can be a non-covalent interaction, including, for example, hydrogen bonding interactions, ionic bonding interactions, hydrophobic interactions, and / or van der Waals interactions. A complex can also include the association of two or more molecules held together by covalent or non-covalent bonds, interactions, or forces. The strength of all non-covalent interactions between a single antigen-binding site on an antibody and a single epitope of a target molecule, such as an antigen, is the affinity of the antibody or functional fragment for that epitope. The dissociation rate (k off ) and association rate (k on ) ratio (k off / k on ) is the dissociation constant K D which is inversely proportional to the affinity. D The lower the value, the higher the affinity of the antibody. D The value of k varies for different antibody-antigen complexes. on and k off The dissociation constant K of the antibodies provided herein depends on both D The affinity of an antibody can be determined by any method provided herein or by any other method known to those skilled in the art. The affinity at one binding site does not necessarily reflect the true strength of the interaction between an antibody and an antigen. When a complex antigen containing multiple repeat antigenic determinants, such as a multivalent antigen, contacts an antibody containing multiple binding sites, the interaction between the antibody and the antigen at one site increases the probability of reaction at a second site. The strength of such multiple interactions between a multivalent antibody and an antigen is called avidity.

[0108] In connection with the antibodies or antigen-binding fragments thereof described herein, terms such as "binds to," "specifically binds to," and similar terms are also used interchangeably herein and refer to binding molecules of an antigen-binding domain, such as polypeptides, that specifically bind to an antigen. Antibodies or antigen-binding fragments that bind to or specifically bind to an antigen may be cross-reactive with related antigens. In certain embodiments, antibodies or antigen-binding fragments that bind to or specifically bind to an antigen do not cross-react with other antigens. Antibodies or antigen-binding fragments that bind to or specifically bind to an antigen can be identified, for example, by immunoassays, Octet®, Biacore®, or other techniques known to those skilled in the art. In some embodiments, an antibody or antigen-binding fragment binds to or specifically binds to an antigen when it binds to the antigen with higher affinity than any cross-reactive antigens, as determined using experimental techniques such as radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA). Typically, a specific or selective response is at least twice the background signal or noise, and may be greater than 10 times the background. For a discussion of binding specificity, see, for example, Fundamental Immunology 332-36 (Paul ed., 2d ed. 1989). In certain embodiments, the degree of binding of an antibody or antigen-binding fragment to a "non-target" protein is less than about 10% of the binding of the binding molecule or antigen-binding domain to its specific target antigen, as determined, for example, by fluorescence-activated cell sorting (FACS) analysis or RIA. Terms such as "specific binding," "specifically binds to," or "specific for" refer to binding that is measurably different from non-specific interactions. Specific binding can be measured, for example, by determining the binding of a molecule compared to the binding of a control molecule, which is generally a molecule of similar structure that does not have binding activity. For example, specific binding can be determined by competition with a control molecule similar to the target, such as an excess of unlabeled target. In this case, specific binding is indicated if the binding of the labeled target to the probe is competitively inhibited by an excess of unlabeled target.Antibodies or antigen-binding fragments that bind to an antigen include those that can bind to the antigen with sufficient affinity so that the binding molecule is useful, for example, as a diagnostic agent in targeting the antigen. In certain embodiments, antibodies or antigen-binding fragments that bind to an antigen have an affinity of less than 1000 nM, less than 800 nM, less than 500 nM, less than 250 nM, less than 100 nM, less than 50 nM, less than 10 nM, less than 5 nM, less than 4 nM, less than 3 nM, less than 2 nM, less than 1 nM, less than 0.9 nM, less than 0.8 nM, less than 0.7 nM, less than 0.6 nM, less than 0.5 nM, less than 0.4 nM a dissociation constant (K) of less than 1000 nM, 800 nM, 500 nM, 250 nM, 100 nM, 50 nM, 10 nM, 5 nM, 4 nM, 3 nM, 2 nM, 1 nM, 0.9 nM, 0.8 nM, 0.7 nM, 0.6 nM, 0.5 nM, 0.4 nM, 0.3 nM, 0.2 nM, or 0.1 nM. D In certain embodiments, the antibody or antigen-binding fragment binds to an epitope of an antigen that is conserved among antigens from different species (e.g., between the human species and the cynomolgus monkey species).

[0109] "Binding affinity" generally refers to the strength of the sum total of non-covalent interactions between a single binding site of a molecule (e.g., a binding protein such as an antibody) and its binding partner (e.g., an antigen). Unless otherwise indicated, as used herein, "binding affinity" refers to the intrinsic binding affinity that reflects a 1:1 interaction between members of a binding pair (e.g., an antibody and an antigen). The affinity of a binding molecule X for its binding partner Y is generally determined by the dissociation constant (K D ) Affinity can be measured by common methods known in the art, including those described herein. Low affinity antibodies generally bind antigens slowly and tend to dissociate easily, whereas high affinity antibodies generally bind antigens faster and tend to remain bound longer. Various methods of measuring binding affinity are known in the art, any of which may be used for purposes of the present disclosure. Specific illustrative embodiments include the following. In one embodiment, "K D " or "KD The "K value" can be measured by assays known in the art, for example, binding assays. D K may be measured, for example, in an RIA performed using the Fab version of the antibody of interest and its antigen (Chen et al., 1999, J. Mol Biol 293:865-81). D or K D The value may also be measured by using biolayer interferometry (BLI) or surface plasmon resonance (SPR) assays, for example by Octet® using an Octet® QK384 system, or by Biacore® using, for example, a Biacore® TM-2000 or Biacore® TM-3000. "On-rate" or "rate of association" or "association rate" or "k" may also be determined using the same biolayer interferometry (BLI) or surface plasmon resonance (SPR) techniques described above, for example, using an Octet® QK384, Biacore® TM-2000, or Biacore® TM-3000 system.

[0110] In certain embodiments, antibodies or antigen-binding fragments can include "chimeric" sequences in which a portion of the heavy and / or light chain is identical to or homologous to corresponding sequences in antibodies from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical to or homologous to corresponding sequences in antibodies from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity (see U.S. Pat. No. 4,816,567 and Morrison et al., 1984, Proc. Natl. Acad. Sci. USA 81:6851-55).

[0111] In certain embodiments, an antibody or antigen-binding fragment can comprise a portion of a "humanized" form of a non-human (e.g., murine) antibody, which is a chimeric antibody comprising a human immunoglobulin (e.g., recipient antibody) in which native CDR residues are replaced by residues from a corresponding CDR (e.g., donor antibody) of a non-human species, such as mouse, rat, rabbit, or non-human primate, containing the desired specificity, affinity, and capacity. In some cases, one or more FR region residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies can comprise residues that are not found in the recipient antibody or the donor antibody. These modifications are made to further refine antibody performance. The heavy or light chain of a humanized antibody can comprise substantially all of at least one or more variable regions, in which all or substantially all of the CDRs correspond to those of a non-human immunoglobulin and all or substantially all of the FRs are those of a human immunoglobulin sequence. In certain embodiments, a humanized antibody comprises at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For further details, see Jones et al., 1986, Nature 321:522-25; Riechmann et al., 1988, Nature 332:323-29; Presta, 1992, Curr. Op. Struct. Biol. 2:593-96; Carter et al., 1992, Proc. Natl. Acad. Sci. USA 89:4285-89; U.S. Patent Nos. 6,800,738, 6,719,971, 6,639,055, 6,407,213, and 6,054,297.

[0112] In certain embodiments, an antibody or antigen-binding fragment can comprise a portion of a "fully human antibody" or "human antibody," which terms are used interchangeably herein and refer to an antibody comprising a human variable region and, for example, a human constant region. In specific embodiments, these terms refer to an antibody comprising variable and constant regions of human origin. A "fully human" antibody, in certain embodiments, can also encompass antibodies that bind to a polypeptide and are encoded by nucleic acid sequences that are naturally occurring somatic variants of human germline immunoglobulin nucleic acid sequences. The term "fully human antibody" includes antibodies comprising variable and constant regions that correspond to human germline immunoglobulin sequences as set forth by Kabat et al. (See Kabat et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, USDapartment of Health and Human Services, NIH Publication No. 91-3242). A "human antibody" is one that has an amino acid sequence that corresponds to that of an antibody produced by a human and / or has been produced using any of the techniques for producing human antibodies. This definition of a human antibody specifically excludes humanized antibodies that comprise non-human antigen-binding residues. Human antibodies can be produced using a variety of techniques known in the art, including phage display libraries (Hoogenboom and Winter, 1991, J. Mol. Biol. 227:381; Marks et al., 1991, J. Mol. Biol. 222:581) and yeast display libraries (Chao et al., 2006, Nature Protocols 1:755-68).Also available for the preparation of human monoclonal antibodies are the methods described by Cole et al., Monoclonal Antibodies and Cancer Therapy 77 (1985), Boerner et al., 1991, J. Immunol. 147(1):86-95, and van Dijk and van de Winkel, 2001, Curr. Opin. Pharmacol. 5:368-74. Human antibodies can be prepared by administering antigen to transgenic animals, e.g., mice, that have been engineered to produce such antibodies in response to antigen challenge but whose endogenous gene loci have been disabled (see, e.g., Jakobovits, 1995, Curr. Opin. Biotechnol. 6(5):561-66; Bruggemann and Taussing, 1997, Curr. Opin. Biotechnol. 8(4):455-58; and U.S. Pat. Nos. 6,075,181 and 6,150,584 regarding XENOMOUSE™ technology). See also, e.g., Li et al., 2006, Proc. Natl. Acad. Sci. USA 103:3557-62 regarding human antibodies generated via human B-cell hybridoma technology.

[0113] In certain embodiments, an antibody or antigen-binding fragment can comprise a portion of a "recombinant human antibody," which phrase includes human antibodies that are prepared, expressed, generated, or isolated by recombinant means, e.g., antibodies expressed using a recombinant expression vector transfected into a host cell, antibodies isolated from a recombinant combinatorial human antibody library, antibodies isolated from an animal (e.g., a mouse or a cow) that is transgenic and / or transchromosomal for human immunoglobulin genes (see, e.g., Taylor, LD et al. (1992) Nucl. Acids Res. 20:6287-6295), or antibodies prepared, expressed, generated, or isolated by any other means, including splicing of human immunoglobulin gene sequences into other DNA sequences. Such recombinant human antibodies can have variable and constant regions derived from human germline immunoglobulin sequences (see Kabat, E.A. et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, USDapartment of Health and Human Services, NIH Publication No. 91-3242). However, in certain embodiments, such recombinant human antibodies have been subjected to in vitro mutagenesis (or, when animals transgenic for human Ig sequences are used, in vivo somatic mutagenesis) such that the amino acid sequences of the VH and VL regions of the recombinant antibodies are derived from and related to human germline VH and VL sequences, but are sequences that may not naturally exist within the human antibody germline repertoire in vivo.

[0114] In certain embodiments, an antibody or antigen-binding fragment can comprise a portion of a "monoclonal antibody," a term used herein to refer to an antibody obtained from a population of substantially homogeneous antibodies, e.g., the individual antibodies comprising the population are identical except for possible minor naturally occurring mutations, and each monoclonal antibody typically recognizes a single epitope on an antigen. In specific embodiments, a "monoclonal antibody," as used herein, is an antibody produced by a single hybridoma or other cell. The term "monoclonal" is not limited to any particular method for producing the antibody. For example, monoclonal antibodies useful in the present disclosure may be prepared by the hybridoma method first described by Kohler et al., 1975, Nature 256:495, or may be produced using recombinant DNA methods in bacterial or eukaryotic animal or plant cells (see, e.g., U.S. Pat. No. 4,816,567). "Monoclonal antibodies" may also be isolated from phage antibody libraries using, for example, the techniques described in Clackson et al., 1991, Nature 352:624-28 and Marks et al., 1991, J. Mol. Biol. 222:581-97. Other methods for preparation of clonal cell lines and the monoclonal antibodies expressed thereby are well known in the art. See, for example, Short Protocols in Molecular Biology (Ausubel et al. eds., 5th ed. 2002).

[0115] A typical four-chain antibody unit is a heterotetrameric glycoprotein composed of two identical light (L) chains and two identical heavy (H) chains. In the case of IgG, the four-chain unit is generally approximately 150,000 daltons. Each L chain is linked to an H chain by one covalent disulfide bond, and the two H chains are linked to each other by one or more disulfide bonds depending on the H chain isotype. Each H and L chain also has regularly spaced intrachain disulfide bridges. Each H chain has a variable domain (VH) at its N-terminus, followed by three constant domains (CH) for each of the α and γ chains, and four CH domains for the μ and ε isotypes. Each L chain has a variable domain (VL) at its N-terminus, followed by a constant domain (CL) at its other end. The VL aligns with the VH, and the CL aligns with the first constant domain (CH1) of the heavy chain. Specific amino acid residues are believed to form an interface between the light chain variable domain and the heavy chain variable domain. The pairing of a VH and VL forms a single antigen-binding site. For the structure and properties of different classes of antibodies, see, for example, Basic and Clinical Immunology 71 (Stites et al. eds., 8th ed. 1994) and Immunobiology (Janeway et al. eds., 5th ed. 2001).

[0116] The term "Fab" or "Fab region" refers to the antibody region that binds to an antigen. Conventional IgGs usually contain two Fab regions, each present in one of the two arms of the Y-shaped IgG structure. Each Fab region typically consists of one variable region and one constant region from each of the heavy and light chains. More specifically, the heavy chain variable and constant regions in the Fab region are the VH and CH1 regions, and the light chain variable and constant regions in the Fab region are the VL and CL regions. The VH, CH1, VL, and CL regions within the Fab region can be arranged in various ways to confer antigen-binding capability according to the present disclosure. For example, as with the Fab region of a conventional IgG, the VH and CH1 regions can be on one polypeptide, while the VL and CL regions can be on separate polypeptides. Alternatively, the VH, CH1, VL, and CL regions can all be on the same polypeptide and oriented in different orders, as described in more detail in the following sections.

[0117] The terms "variable region," "variable domain," "V region," or "V domain" refer to the portion of an antibody light or heavy chain, generally located at the amino terminus of the light or heavy chain, approximately 120 to 130 amino acids in length for heavy chains and approximately 100 to 110 amino acids in length for light chains, that is used to determine the binding and specificity of each particular antibody for its specific antigen. The variable region of a heavy chain may be referred to as "VH." The variable region of a light chain may be referred to as "VL." The term "variable" refers to the fact that certain segments of the variable region vary significantly in sequence among antibodies. The V region mediates antigen binding and determines the specificity of a particular antibody for a particular antigen. However, variability is not evenly distributed across the 110-amino acid span of the variable region. Instead, the V region consists of less variable (e.g., relatively invariant) stretches of approximately 15 to 30 amino acids called framework regions (FRs) separated by shorter regions of greater variability (e.g., extreme variability) called "hypervariable regions," each approximately 9 to 12 amino acids in length. The heavy and light chain variable regions each contain four FRs, primarily adopting a β-sheet configuration, connected by three hypervariable regions that form loops that connect and, in some cases, form part of the β-sheet structure. The hypervariable regions of each chain are held in close proximity by the FRs and, together with the hypervariable regions of the other chain, contribute to the formation of the antigen-binding site of antibodies (see, e.g., Kabat et al., Sequences of Proteins of Immunological Interest (5th ed. 1991)). The constant regions are not directly involved in binding the antibody to the antigen, but exhibit various effector functions, such as antibody participation in antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC). The sequences of the variable regions vary significantly among different antibodies. In a specific embodiment, the variable regions are human variable regions.

[0118] The terms "variable region residue numbering according to Kabat" or "amino acid position numbering as in Kabat," and variations thereof, refer to the numbering system used for the heavy or light chain variable regions of the compilation of antibodies in Kabat et al., supra. Using this numbering system, the actual linear amino acid sequence may contain fewer or additional amino acids corresponding to shortening of, or insertions into, FRs or CDRs of the variable domain. For example, a heavy chain variable domain may contain a single amino acid insertion after residue 52 (residue 52a according to Kabat) and three inserted residues after residue 82 (e.g., residues 82a, 82b, and 82c according to Kabat). The Kabat numbering of residues can be determined for a given antibody by alignment of the antibody's sequence with the "standard" Kabat numbered sequence at the regions of homology. The Kabat numbering system is commonly used when referring to residues in the variable domain (approximately residues 1-107 of the light chain and residues 1-113 of the heavy chain) (e.g., Kabat et al., supra). The "EU numbering system" or "EU index" is commonly used when referring to residues within the immunoglobulin heavy chain constant region (e.g., the EU index reported in Kabat et al., supra). "EU index as in Kabat" refers to the residue numbering of the human IgG1 EU antibody. Other numbering systems are described, for example, by AbM, Chothia, Contact, IMGT, and AHon.

[0119] The term "heavy chain," when used in reference to an antibody, refers to a polypeptide chain of approximately 50 to 70 kDa, the amino-terminal portion of which contains a variable region of approximately 120 to 130 or more amino acids, and the carboxy-terminal portion of which contains a constant region. The constant region can be one of five different types (e.g., isotypes), designated alpha (α), delta (δ), epsilon (ε), gamma (γ), and mu (μ), based on the amino acid sequence of the heavy chain constant region. Different heavy chains vary in size, with α, δ, and γ containing approximately 450 amino acids, and μ and ε containing approximately 550 amino acids. When combined with light chains, these different types of heavy chains give rise to five well-known classes (e.g., isotypes) of antibodies: IgA, IgD, IgE, IgG, and IgM, respectively, including the four subclasses of IgG: IgG1, IgG2, IgG3, and IgG4.

[0120] The term "light chain," when used in reference to an antibody, refers to a polypeptide chain of approximately 25 kDa, the amino-terminal portion of which contains a variable region of about 100 to about 110 or more amino acids, and the carboxy-terminal portion of which contains a constant region. The approximate length of a light chain is 211 to 217 amino acids. There are two different types, called kappa (κ) or lambda (λ), based on the amino acid sequence of the constant domain.

[0121] As used herein, the terms "hypervariable region," "HVR," "complementarity-determining region," and "CDR" are used interchangeably. "CDR" refers to one of the three hypervariable regions (H1, H2, or H3) within the non-framework region of an immunoglobulin (Ig or antibody) VH β-sheet framework, or one of the three hypervariable regions (L1, L2, or L3) within the non-framework region of an antibody VL β-sheet framework. Thus, CDRs are variable region sequences interspersed within framework region sequences.

[0122] CDR regions are well known to those skilled in the art and are defined by well-known numbering systems. For example, Kabat complementarity-determining regions (CDRs) are based on sequence variability and are the most commonly used (see, e.g., Kabat et al., supra). Instead, Chothia refers to the position of the structural loop (see, e.g., Chothia and Lesk, 1987, J. Mol. Biol. 196:901-17). When numbered using Kabat's numbering rules, the end of the Chothia CDR-H1 loop varies between H32 and H34 depending on the length of the loop (this is because the Kabat numbering scheme places insertions at H35A and H35B; if neither 35A nor 35B is present, the loop ends at 32; if only 35A is present, the loop ends at 33; if both 35A and 35B are present, the loop ends at 34). AbM hypervariable regions represent a compromise between Kabat CDRs and Chothia structural loops and are used by Oxford Molecular's AbM antibody modeling software (see, e.g., Antibody Engineering Vol. 2 (Kontermann and Dubel eds., 2nd ed. 2010)). The "contact" hypervariable regions are based on the analysis of available complex crystal structures. Another universal numbering system that has been developed and widely adopted is the ImMunoGeneTics (IMGT) Information System® (Lafranc et al., 2003, Dev. Comp. Immunol. 27(1):55-77). IMGT is an integrated information system specialized for immunoglobulins (IGs), T cell receptors (TCRs), and major histocompatibility complexes (MHCs) of humans and other vertebrates. Herein, CDRs are referred to both with respect to amino acid sequence and location within the light or heavy chain. Because the "locations" of CDRs within the structure of immunoglobulin variable domains are conserved across species and reside within structures called loops, CDR and framework residues are readily identified by using a numbering system that aligns variable domain sequences according to structural features.This information can be used to graft and replace CDR residues from one species' immunoglobulin onto an acceptor framework, typically from a human antibody. An additional numbering system (AHon) has been developed by Honegger and Pluckthun, 2001, J. Mol. Biol. 309:657-70. For example, the correspondence between numbering systems, including the Kabat numbering and the IMGT-specific numbering system, is well known to those of skill in the art (see, e.g., Kabat, supra; Chothia and Lesk, supra; Martin, supra; Lefranc et al., supra). Residues from each of these hypervariable regions or CDRs are shown in Table 1 below.

[0123] (Table 1) TIFF2025501978000005.tif77137

[0124] The boundaries of a given CDR may vary depending on the scheme used for identification. Thus, unless otherwise specified, the terms "CDR" and "complementarity determining region" of a given antibody or region thereof, e.g., a variable region, and individual CDRs of an antibody or region thereof (e.g., "CDR-H1, CDR-H2"), should be understood to encompass the complementarity determining regions defined by any of the known schemes described hereinabove. In some cases, a scheme for identifying a particular CDR or CDRs is specified, such as CDRs defined by the Kabat, Chothia, or Contact methods. In other cases, the specific amino acid sequences of the CDRs are given.

[0125] The hypervariable regions may comprise "extended hypervariable regions" as follows: 24-36 or 24-34 (L1), 46-56 or 50-56 (L2), and 89-97 or 89-96 (L3) in VL, and 26-35 or 26-35A (H1), 50-65 or 49-65 (H2), and 93-102, 94-102, or 95-102 (H3) in VH.

[0126] The term "constant region" or "constant domain" refers to the carboxy-terminal portions of the light and heavy chains that are not directly involved in binding an antibody to an antigen but exhibit various effector functions, such as interaction with Fc receptors. This term refers to the portion of an immunoglobulin molecule that contains a more conserved amino acid sequence compared to other portions of the immunoglobulin, the variable region, which contains the antigen-binding site. The constant region may include the CH1, CH2, and CH3 regions of the heavy chain and the CL region of the light chain.

[0127] The term "framework" or "FR" refers to variable region residues that flank the CDRs. FR residues are present, for example, in chimeric, humanized, human, domain antibodies, diabodies, linear antibodies, and bispecific antibodies. FR residues are variable domain residues other than hypervariable region or CDR residues.

[0128] The term "Fc region" herein is used to define the C-terminal region of an immunoglobulin heavy chain, including, for example, native-sequence Fc regions, recombinant Fc regions, and variant Fc regions. Although the boundaries of the Fc region of an immunoglobulin heavy chain can vary, the human IgG heavy chain Fc region is often defined to extend from the amino acid residue at position Cys226 or the amino acid residue at position Pro230 to its carboxyl terminus. The C-terminal lysine of the Fc region (residue 447 according to the EU numbering system) can be removed, for example, during antibody production or purification, or by recombinantly engineering the nucleic acid encoding the antibody heavy chain. Thus, a composition of intact antibodies can include antibody populations in which all K447 residues have been removed, antibody populations in which the K447 residue has not been removed, and antibody populations containing a mixture of antibodies with and without the K447 residue. A "functional Fc region" possesses the "effector functions" of a native-sequence Fc region. Exemplary "effector functions" include C1q binding, CDC, Fc receptor binding, ADCC, phagocytosis, down-regulation of cell surface receptors (e.g., B cell receptors), and the like. Such effector functions generally require an Fc region in combination with a binding region or domain (e.g., an antibody variable region or domain) and can be assessed using a variety of assays known to those skilled in the art. A "variant Fc region" comprises an amino acid sequence that differs from that of a native-sequence Fc region by at least one amino acid modification (e.g., substitution, addition, or deletion). In certain embodiments, a variant Fc region has at least one amino acid substitution compared to a native-sequence Fc region or the Fc region of a parent polypeptide, e.g., from about 1 to about 10 amino acid substitutions, or from about 1 to about 5 amino acid substitutions in the native-sequence Fc region or the Fc region of a parent polypeptide. The variant Fc region herein may have at least about 80% homology to a native sequence Fc region and / or the Fc region of a parent polypeptide, or at least about 90% homology thereto, for example, at least about 95% homology thereto.

[0129] As used herein, "epitope" is a term of art that refers to a localized region of an antigen to which a binding molecule (e.g., an antibody) can specifically bind. An epitope can be a linear epitope, a conformational epitope, a non-linear epitope, or a discontinuous epitope. In the case of a polypeptide antigen, for example, an epitope can be contiguous amino acids of the polypeptide (a "linear" epitope), or it can include amino acids from two or more discontinuous regions of the polypeptide (a "conformational," "non-linear," or "discontinuous" epitope). In general, those skilled in the art will understand that a linear epitope may or may not depend on secondary, tertiary, or quaternary structure. For example, in some embodiments, a binding molecule binds to a group of amino acids, regardless of whether they are folded into a native three-dimensional protein structure. In other embodiments, the binding molecule requires the amino acid residues that make up the epitope to adopt a particular conformation (eg, a bend, twist, turn, or fold) in order to recognize and bind to the epitope.

[0130] The terms "polypeptide," "peptide," and "protein" are used interchangeably herein to refer to polymers of amino acids of any length. The polymers may be linear or branched, may comprise modified amino acids, and may be interrupted by non-amino acids. The term also encompasses amino acid polymers that are modified, naturally or by intervention, for example, by disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation or modification. Also included within the definition are polypeptides containing one or more analogs of an amino acid, including, but not limited to, unnatural amino acids, as well as other modifications known in the art. Because the polypeptides of the present disclosure may be based on antibodies or other members of the immunoglobulin superfamily, in certain embodiments, it is understood that a "polypeptide" can occur as a single chain or as two or more associated chains.

[0131] As used herein, the term "pharmaceutically acceptable" means approved by a regulatory agency of a federal or state government or listed in the United States Pharmacopoeia, the European Pharmacopoeia, or other generally recognized pharmacopoeias for use in animals, and more particularly in humans.

[0132] "Excipient" means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, solvent, or encapsulating material. Excipients include, for example, encapsulating materials or additives such as absorption enhancers, antioxidants, binders, buffers, carriers, coating agents, colorants, diluents, disintegrants, emulsifiers, bulking agents, fillers, flavoring agents, humectants, lubricants, perfumes, preservatives, propellants, releasing agents, sterilizing agents, sweeteners, solubilizing agents, wetting agents, and mixtures thereof. The term "excipient" can also refer to a diluent, adjuvant (e.g., Freund's adjuvant (complete or incomplete)), or vehicle.

[0133] In one embodiment, each component is "pharmaceutically acceptable" in the sense of being compatible with the other components of the pharmaceutical formulation and suitable for use in contact with the tissues or organs of humans and animals without undue toxicity, irritation, allergic response, immunogenicity, or other problems or complications, commensurate with a reasonable benefit / risk ratio. See, e.g., Lippincott Williams & Wilkins: Philadelphia, PA, 2005; Handbook of Pharmaceutical Excipients, 6th ed., Rowe et al., Eds., The Pharmaceutical Press and the American Pharmaceutical Association: 2009; Handbook of Pharmaceutical Additives, 3rd ed., Ash and Ash Eds., Gower Publishing Company: 2007; Pharmaceutical Preformulation and Formulation, 2nd ed., Gibson Ed., CRC Press LLC: Boca Raton, FL, 2009. In some embodiments, a pharmaceutically acceptable excipient is nontoxic to cells or mammals exposed thereto at the dosages and concentrations employed. In some embodiments, the pharmaceutically acceptable excipient is an aqueous pH buffered solution.

[0134] The abbreviation "MMAE" refers to monomethyl auristatin E.

[0135] Unless the context dictates otherwise, a hyphen (-) designates the point of attachment to the pendant molecule.

[0136] The term "chemotherapeutic agent" refers to any chemical compound that is effective in inhibiting tumor growth. Non-limiting examples of chemotherapeutic agents include alkylating agents, such as nitrogen mustard, ethyleneimine compounds, and alkylsulfonates; antimetabolites, such as folic acid, purine, or pyrimidine antagonists; mitotic inhibitors, such as antitubulin agents, such as derivatives of vinca alkaloids, auristatins, and podophyllotoxins; cytotoxic antibiotics; compounds that damage or interfere with DNA expression or replication, such as DNA minor groove binders; and growth factor receptor antagonists. In addition, chemotherapeutic agents include cytotoxic agents (as defined herein), antibodies, biological molecules, and small molecules.

[0137] As used herein, the term "conservative substitution" is known to those skilled in the art and generally refers to an amino acid substitution that may be made without altering the biological activity of the resulting molecule. Those skilled in the art generally recognize that single amino acid substitutions in non-essential regions of a polypeptide do not substantially alter biological activity (see, for example, Watson et al., MOLECULAR BIOLOGY OF THE GENE, The Benjamin / Cummings Pub. Co., p. 224 (4th Edition 1987)). Such exemplary substitutions are preferably made in accordance with the substitutions shown in Tables 2 and 3. For example, such changes include substituting any of isoleucine (I), valine (V), and leucine (L) with any other of these hydrophobic amino acids; substituting glutamic acid (E) with aspartic acid (D) and vice versa; substituting asparagine (N) with glutamine (Q) and vice versa; and substituting threonine (T) with serine (S) and vice versa. Other substitutions may also be considered conservative, depending on the particular amino acid's environment and its role in the protein's three-dimensional structure. For example, glycine (G) and alanine (A) can often be interchangeable, as can alanine (A) and valine (V). Methionine (M), which is relatively hydrophobic, can often be interchanged with leucine and isoleucine, and sometimes with valine. Lysine (K) and arginine (R) are often interchangeable in positions where the important feature of an amino acid residue is its charge, and the different pKs of these two amino acid residues are not important. Still other changes may be considered "conservative" in certain circumstances (see, e.g., Table 3 herein; pages 13-15 of "Biochemistry," 2nd ed. Lubert Stryer ed. (Stanford University); Henikoff et al., PNAS 1992 Vol. 89 10915-10919; Lei et al., J. Biol. Chem 1995 May 19;270(20):11882-11886). Other substitutions are permissible and may be determined empirically or in accordance with known conservative substitutions.

[0138] Table 2: Amino acid abbreviations TIFF2025501978000006.tif144158

[0139] Table 3: Amino acid substitution or similarity matrix TIFF2025501978000007.tif150165

[0140] The term "homology" or "homologous" is intended to refer to sequence similarity between two polynucleotides or two polypeptides. Similarity can be determined by comparing positions in each sequence that can be aligned for comparison purposes. If a given position in two polypeptide sequences is not identical, the similarity or conservation of that position can be determined by assessing the amino acid similarity at that position, for example, according to Table 3. The degree of similarity between sequences is a function of the number of matching or homologous positions shared by the sequences. Alignment of two sequences to determine percent sequence similarity can be performed using software programs known in the art, such as those described in Ausubel et al., Current Protocols in Molecular Biology, John Wiley and Sons, Baltimore, MD (1999). Preferably, default parameters are used for alignment, an example of which is provided below. One alignment program known in the art that can be used is BLAST, set to default parameters. In particular, the programs are BLASTN and BLASTP, using the following default parameters: Genetic code = standard; filter = none; strand = both; cutoff = 60; expectation = 10; matrix = BLOSUM62; description = 50 sequences; sort order = high score; database = non-redundant, GenBank + EMBL + DDBJ + PDB + GenBank CDS translation + SwissProtein + SPupdate + PIR. Details of these programs can be found at the National Center for Biotechnology Information.

[0141] The term "homolog" of a given amino acid or nucleic acid sequence is intended to indicate a corresponding sequence of the "homolog" having substantial identity or homology to the given amino acid or nucleic acid sequence.

[0142] The determination of percent identity between two sequences (e.g., amino acid sequences or nucleic acid sequences) can be accomplished using a mathematical algorithm. A preferred, non-limiting example of a mathematical algorithm utilized for comparing two sequences is the algorithm of Karlin and Altschul, 1990, Proc. Natl. Acad. Sci. USA 87:2264 2268, modified as in Karlin and Altschul, 1993, Proc. Natl. Acad. Sci. USA 90:5873 5877. Such an algorithm is incorporated into the NBLAST and XBLAST programs of Altschul et al., 1990, J. Mol. Biol. 215:403. BLAST nucleotide searches can be performed, for example, using the NBLAST nucleotide program parameters set to score=100 and word length=12 to obtain nucleotide sequences homologous to the nucleic acid molecules described herein. BLAST protein searches can be performed using, for example, XBLAST program parameters set to score 50 and word length = 3 to obtain amino acid sequences homologous to the protein molecules described herein. To obtain gapped alignments for comparison, Gapped BLAST can be utilized as described in Altschul et al., 1997, Nucleic Acids Res. 25:3389-3402. Alternatively, PSI BLAST can be used to perform an iterated search that detects distant relationships between molecules (ibid.). When utilizing BLAST, Gapped BLAST, and PSI Blast programs, the default parameters of the respective programs (e.g., XBLAST and NBLAST) can be used (see, e.g., the National Center for Biotechnology Information (NCBI) on the World Wide Web at ncbi.nlm.nih.gov). Another non-limiting example of a mathematical algorithm utilized for sequence comparison is the algorithm of Myers and Miller, 1988, CABIOS 4:11-17.Such an algorithm is incorporated into the ALIGN program (version 2.0), which is part of the GCG sequence alignment software package. When utilizing the ALIGN program for comparing amino acid sequences, a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4 can be used.

[0143] The percent identity between two sequences can be determined using techniques similar to those described above, with or without allowing gaps. In calculating percent identity, typically only exact matches are counted.

[0144] The term "cytotoxic agent" refers to a substance that inhibits or prevents the development, activity, function, and / or causes destruction of cells. This term is intended to include radioisotopes, chemotherapeutic agents, and toxins, such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant, or animal origin, including fragments and / or variants. Examples of cytotoxic agents include auristatins (e.g., auristatin E, auristatin F, MMAE, and MMAF), aureomycin, maytansinoids, ricin, ricin A chain, combrestatins, duocarmycins, dolastatins, doxorubicin, daunorubicin, taxol, cisplatin, cc1065, ethidium bromide, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicine, dihydroxyanthracin dione, actinomycin, diphtheria toxin, Pseudomonas exotoxin (PE) A, PE40, abrin, abrin A chain, modeccin A chain, α-sarcin, gelonin, mitogenin, restrictocin, phenomycin, enomycin, curicin, crotin, calicheamicin, soapwort (Sapaonaria officinalis) inhibitors, and glucocorticoids and other chemotherapeutic agents, as well as At 211 , I 131 , I 125 , Y 90 ,Re 186 ,Re 188 , Sm 153 , Bi 212 or Bi213 , P 32 and radioactive isotopes of Lu, including Lu 177. The antibody may also be conjugated to an anti-cancer prodrug-activating enzyme capable of converting the prodrug to its active form.

[0145] As used herein, the term "effective amount" or "therapeutically effective amount" refers to an amount of a binding molecule (e.g., an antibody) or pharmaceutical composition provided herein sufficient to result in a desired outcome.

[0146] The terms "subject" and "patient" may be used interchangeably. As used herein, in certain embodiments, a subject is a mammal, such as a non-primate (e.g., cows, pigs, horses, cats, dogs, rats, etc.) or a primate (e.g., monkeys and humans). In certain embodiments, a subject is a human. In one embodiment, a subject is a mammal, e.g., a human, who has been diagnosed with a condition or disorder. In another embodiment, a subject is a mammal, e.g., a human, who is at risk of developing a condition or disorder.

[0147] "Administering" or "administration" refers to the act of injecting or otherwise physically delivering a substance present outside the body to a patient, such as by mucosal, intradermal, intravenous, intramuscular delivery, and / or any other physical delivery method described herein or known in the art.

[0148] As used herein, the terms "treat," "treatment," and "treating" refer to a reduction or amelioration of the progression, severity, and / or duration of a disease or condition resulting from the administration of one or more therapies. Treatment may be determined by assessing whether there is a reduction, alleviation, and / or relief of one or more symptoms associated with the underlying disease, such that an improvement is observed for the patient, even though the patient may still be suffering from the underlying disease. The term "treating" includes both management and amelioration of disease. The terms "manage," "managing," and "management" refer to the beneficial effects a subject derives from a therapy that does not necessarily result in a cure of the disease.

[0149] The terms "prevent," "preventing," and "prevention" refer to reducing the likelihood of the occurrence (or recurrence) of a disease, disorder, condition, or associated symptom(s) (e.g., cancer).

[0150] The terms "cancer" or "cancer cells" are used herein to refer to tissue or cells found in neoplasms that have characteristics that distinguish them from normal tissue or tissue cells. Such characteristics include, but are not limited to, the degree of anaplasia, irregular shape, unclear cell outlines, nuclear size, changes in nuclear or cytoplasmic structure, other phenotypic changes, the presence of cellular proteins indicative of cancer or precancerous conditions, an increased number of mitoses, and the ability to metastasize. Words related to "cancer" include carcinoma, sarcoma, tumor, epithelioma, leukemia, lymphoma, polyp, and scirrhous carcinoma, transformation, neoplasia, etc.

[0151] As used herein, "locally advanced" cancer refers to cancer that has spread from where it began to nearby tissues or lymph nodes.

[0152] As used herein, "metastatic" cancer refers to cancer that has spread from where it began to another part of the body.

[0153] As used herein, "substantially as effective" means that the treatment is at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% as effective as SOC therapy.

[0154] The terms "about" and "approximately" mean within 20%, within 15%, within 10%, within 9%, within 8%, within 7%, within 6%, within 5%, within 4%, within 3%, within 2%, within 1%, or less of a given value or range.

[0155] As used in this disclosure and claims, the singular forms "a," "an," and "the" include the plural forms unless the context clearly dictates otherwise.

[0156] Whenever an embodiment is described herein using the term "comprising," it is understood that other similar embodiments described in terms of "consisting of" and / or "consisting essentially of" are also provided. Also, whenever an embodiment is described herein using the phrase "consisting essentially of," it is understood that other similar embodiments described in terms of "consisting of" are also provided.

[0157] The term "and / or" as used herein in phrases such as "A and / or B" is intended to include both A and B; A or B; A (alone); and B (alone). Similarly, when used in phrases such as "A, B, and / or C," the term "and / or" is intended to encompass each of the following embodiments: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (only); B (only); and C (only).

[0158] The term "variant" refers to a molecule that exhibits variation from a described type or reference, such as a protein with one or more different amino acid residues at the corresponding position(s) of a specifically described protein (e.g., the 191P4D12 protein shown in Figure 1A). Analogs are examples of variant proteins. Splice isoforms and single nucleotide polymorphisms (SNPs) are further examples of variants.

[0159] "191P4D12 proteins" and / or "191P4D12-related proteins" of the present disclosure include those specifically identified herein (see FIG. 1A), as well as allelic variants, conservatively substituted variants, analogs, and homologs, which can be isolated / produced and characterized without undue experimentation according to methods outlined herein or readily available in the art. Also included are fusion proteins that combine portions of different 191P4D12 proteins or fragments thereof, as well as fusion proteins of a 191P4D12 protein and a heterologous polypeptide. Such 191P4D12 proteins are collectively referred to as 191P4D12-related proteins, proteins of the present disclosure, or 191P4D12. The term "191P4D12-related protein" refers to a 191P4D12-related protein that is 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, or more than 25 amino acids, or at least 30, at least 35, at least 40, at least 45, at least 50, at least 55, at least 60, at least 65, at least 70, at least 80, at least 85, at least 90, at least 95, at least 100, at least 105, at least 110, at least 115, at least 120, "191P4D12" refers to a polypeptide fragment or 191P4D12 protein sequence of at least 125, at least 130, at least 135, at least 140, at least 145, at least 150, at least 155, at least 160, at least 165, at least 170, at least 175, at least 180, at least 185, at least 190, at least 195, at least 200, at least 225, at least 250, at least 275, at least 300, at least 325, at least 330, at least 335, at least 339 or more amino acids. The term "191P4D12" is used interchangeably with Nectin-4.

[0160] 5.2 Cancer treatment methods Urothelial carcinoma and bladder cancer, including muscle-invasive urothelial carcinoma (MIUC) or muscle-invasive bladder cancer (MIBC), in patients who are cisplatin-ineligible are particularly difficult to treat. Cisplatin-ineligible MIUC and MIBC patients are typically somewhat frail and suffer from multiple comorbidities beyond urothelial carcinoma / bladder cancer. As noted in the background section above, there are currently no standard neoadjuvant treatment options for cisplatin-ineligible patients with MIBC or MIUC. Instead, these patients are typically treated directly with surgery. The present disclosure provides effective and safe neoadjuvant and perioperative methods for treating urothelial carcinoma and / or bladder cancer, particularly patients with muscle-invasive urothelial carcinoma (MIUC) or muscle-invasive bladder cancer (MIBC) who are cisplatin-ineligible in this setting.

[0161] For example, the results described herein demonstrate that, in some embodiments, the efficacy of treatment of a subject with an effective amount of an ADC as described herein is similar to the efficacy of treatment observed in cisplatin-eligible patients (i.e., healthier patients) treated with standard of care (SOC) therapy used to treat cisplatin-eligible subjects with MIUC or MIBC. The results described herein further demonstrate that, in certain embodiments, even after multiple cycles of treatment with an ADC described herein, the subject remains suitable for radical cystectomy and pelvic lymph node dissection (RC+PLND) surgery, i.e., the method is well tolerated such that the subject remains healthy enough to undergo surgery after neoadjuvant treatment. Finally, the results described herein further demonstrate that, in certain embodiments, the number of cycles of treatment with an ADC described herein administered to a cisplatin-ineligible subject can be equal to or less than the number of cycles of SOC therapy used to treat cisplatin-eligible subjects (i.e., healthier subjects) with MIUC or MIBC. Prior to the results described herein, there was considerable uncertainty as to whether the methods provided herein would be effective and tolerable, given that this patient population has historically proven very difficult to treat and there is currently no standard of care for cis-incompetent MIBC and MIBC patients. Thus, the levels of efficacy and safety obtained using the methods described herein were particularly remarkable and surprising.

[0162] 5.2.1 Cancer treatment methods for general and selected patients Provided herein are methods for the treatment of various cancers in subjects, including subjects with urothelial carcinoma, using antibody drug conjugates (ADCs) that bind to 191P4D12 (Nectin-4).

[0163] In one aspect, provided herein are methods for treating cancer in a human subject using an ADC that binds to 191P4D12 (Nectin-4). In some embodiments, the cancer is muscle-invasive urothelial carcinoma (MIUC) or muscle-invasive bladder cancer (MIBC). In some embodiments, the method comprises administering to the subject an effective amount of an antibody-drug conjugate (ADC). In some embodiments, the ADC comprises an antibody or antigen-binding fragment thereof that binds to 191P4D12 (Nectin-4). In some embodiments, the ADC comprises an antibody or antigen-binding fragment thereof that binds to 191P4D12 (Nectin-4) conjugated to one or more units of a cytotoxic or cytostatic agent. In some embodiments, the ADC comprises an antibody or antigen-binding fragment thereof that binds to 191P4D12 (Nectin-4) conjugated to one or more units of an auristatin agent. In some embodiments, the ADC comprises an antibody or antigen-binding fragment thereof that binds to 191P4D12 (Nectin-4) conjugated to one or more units of monomethyl auristatin E (MMAE). Additional examples of suitable ADCs that can be used in the methods disclosed herein are provided in Section 5.3.2. In some embodiments, the subject is ineligible to receive cisplatin therapy (cisplatin-ineligible). In some embodiments, the pathological downstaging rate (pDSR) is at least 50%. In some embodiments, the pathological downstaging rate (pDSR) is at least 50%. In some embodiments, an effective amount of the ADC is administered to the subject on days 1 and 8 of every 3-week cycle for a total of 3 cycles. In some embodiments, the effective amount of the ADC is about 1.25 mg / kg of subject body weight. In some embodiments, the effective amount of the ADC is about 1.0 mg / kg of subject body weight.

[0164] In some embodiments, after the administering step, the subject undergoes radical cystectomy and pelvic lymph node dissection (RC+PLND) as part of the treatment for cancer (i.e., the treatment for cancer further comprises radical cystectomy and pelvic lymph node dissection (RC+PLND)). In some embodiments, the method comprises administering to the subject two or three cycles of an effective amount of an antibody drug conjugate (ADC). In some embodiments, the method comprises administering to the subject two cycles of an effective amount of an antibody drug conjugate (ADC). In some embodiments, the method comprises administering to the subject three cycles of an effective amount of an antibody drug conjugate (ADC). In some embodiments, the subject has urothelial carcinoma or bladder cancer. In some embodiments, the subject has muscle-invasive urothelial carcinoma (MIUC) or muscle-invasive bladder cancer (MIBC). In some embodiments, the subject is ineligible to receive cisplatin therapy (cisplatin-ineligible).

[0165] In one aspect, the method comprises administering to a subject three cycles of an effective amount of an antibody drug conjugate (ADC), wherein after the administering step, the subject undergoes radical cystectomy and pelvic lymph node dissection (RC+PLND) as part of treatment for cancer, the ADC comprises an antibody or antigen-binding fragment thereof that binds to 191P4D12 (Nectin-4) conjugated to one or more units of monomethyl auristatin E (MMAE), the subject has muscle-invasive urothelial carcinoma (MIUC) or muscle-invasive bladder cancer (MIBC), and the subject is ineligible to receive cisplatin treatment (cisplatin-ineligible).

[0166] In one aspect, the method includes: (i) administering to the subject an effective amount of an ADC on days 1 and 8 of every 3-week cycle, wherein the ADC comprises an antibody or antigen-binding fragment thereof that binds to 191P4D12 (Nectin-4) conjugated to one or more units of monomethyl auristatin E (MMAE); (ii) performing radical cystectomy and pelvic lymph node dissection (RC+PLND) on the subject; and (iii) approximately 8 weeks after step (ii), administering an effective amount of the ADC on days 1 and 8 of every 3-week cycle for a total of 6 cycles, wherein the subject is ineligible to receive cisplatin therapy (cisplatin-ineligible). In some embodiments, the effective amount of the ADC is 1.25 mg / kg. In some embodiments, in step (iii), the effective amount of the ADC is administered for a total of 5 cycles. In some embodiments, in step (iii), the effective amount of the ADC is administered for a total of 4 cycles. In some embodiments, in step (iii), an effective amount of the ADC is administered for a total of 3 cycles. In some embodiments, in step (iii), an effective amount of the ADC is administered for a total of 2 cycles. In some embodiments, in step (iii), an effective amount of the ADC is administered for a total of 1 cycle.

[0167] In one aspect, the method includes: (i) administering to the subject an effective amount of an ADC on days 1 and 8 of a three-week cycle, wherein the ADC comprises an antibody or antigen-binding fragment thereof that binds to 191P4D12 (Nectin-4) conjugated to one or more units of monomethyl auristatin E (MMAE), and wherein after the administering step, the subject undergoes RC+PLND as part of a treatment for cancer; and (ii) after the subject has undergone RC+PLND, approximately 8 weeks after the subject has undergone RC+PLND, administering to the subject an effective amount of an ADC on days 1 and 8 of a three-week cycle for a total of six cycles, wherein the subject is ineligible to receive cisplatin therapy (cisplatin-ineligible). In some embodiments, the effective amount of the ADC is 1.25 mg / kg. In some embodiments, in step (ii), the effective amount of the ADC is administered for a total of five cycles. In some embodiments, in step (ii), the effective amount of the ADC is administered for a total of four cycles. In some embodiments, in step (ii), an effective amount of the ADC is administered for a total of 3 cycles. In some embodiments, in step (ii), an effective amount of the ADC is administered for a total of 2 cycles. In some embodiments, in step (ii), an effective amount of the ADC is administered for a total of 1 cycle.

[0168] In some embodiments of the methods provided herein, the human subject undergoes RC+PLND about 4 to about 12 weeks after the ADC is administered to the human subject. In some embodiments, the human subject undergoes RC+PLND about 5 to about 12 weeks after the ADC is administered to the human subject. In some embodiments, the human subject undergoes RC+PLND about 6 to about 12 weeks after the ADC is administered to the human subject. In some embodiments, the human subject undergoes RC+PLND about 7 to about 12 weeks after the ADC is administered to the human subject. In some embodiments, the human subject undergoes RC+PLND about 8 to about 12 weeks after the ADC is administered to the human subject. In some embodiments, the human subject undergoes RC+PLND about 9 to about 12 weeks after the ADC is administered to the human subject. In some embodiments, the human subject undergoes RC+PLND about 10 to about 12 weeks after the ADC is administered to the human subject. In some embodiments, the human subject undergoes RC+PLND about 11 to about 12 weeks after the ADC is administered to the human subject. In some embodiments, the human subject undergoes RC+PLND about 4 to about 11 weeks after the ADC is administered to the human subject. In some embodiments, the human subject undergoes RC+PLND about 4 to about 10 weeks after the ADC is administered to the human subject. In some embodiments, the human subject undergoes RC+PLND about 4 to about 9 weeks after the ADC is administered to the human subject. In some embodiments, the human subject undergoes RC+PLND about 4 to about 8 weeks after the ADC is administered to the human subject. In some embodiments, the human subject undergoes RC+PLND about 4 to about 7 weeks after the ADC is administered to the human subject. In some embodiments, the human subject undergoes RC+PLND about 4 to about 6 weeks after the ADC is administered to the human subject. In some embodiments, the human subject undergoes RC+PLND about 4 to about 5 weeks after the ADC is administered to the human subject. In some embodiments, the human subject undergoes RC+PLND about 6 to about 8 weeks after the ADC is administered to the human subject. In some embodiments, the human subject undergoes RC+PLND about 7 to about 9 weeks after the ADC is administered to the human subject.In some embodiments, the human subject undergoes RC+PLND about 8 to about 10 weeks after the ADC is administered to the human subject. In some embodiments, the human subject undergoes RC+PLND about 9 to about 11 weeks after the ADC is administered to the human subject.

[0169] In some embodiments of the methods provided herein, the human subject undergoes RC+PLND about 4 weeks after the ADC is administered to the human subject. In some embodiments, the human subject undergoes RC+PLND about 5 weeks after the ADC is administered to the human subject. In some embodiments, the human subject undergoes RC+PLND about 6 weeks after the ADC is administered to the human subject. In some embodiments, the human subject undergoes RC+PLND about 7 weeks after the ADC is administered to the human subject. In some embodiments, the human subject undergoes RC+PLND about 8 weeks after the ADC is administered to the human subject. In some embodiments, the human subject undergoes RC+PLND about 9 weeks after the ADC is administered to the human subject. In some embodiments, the human subject undergoes RC+PLND about 10 weeks after the ADC is administered to the human subject. In some embodiments, the human subject undergoes RC+PLND about 11 weeks after the ADC is administered to the human subject. In some embodiments, the human subject undergoes RC+PLND about 12 weeks after the ADC is administered to the human subject.

[0170] In some embodiments, the method further comprises step (b) of performing a radical cystectomy and pelvic lymph node dissection (RC+PLND) on the subject. In some embodiments, step (b) is performed about 4 to about 12 weeks after step (a). In some embodiments, step (b) is performed about 5 to about 12 weeks after step (a). In some embodiments, step (b) is performed about 6 to about 12 weeks after step (a). In some embodiments, step (b) is performed about 7 to about 12 weeks after step (a). In some embodiments, step (b) is performed about 8 to about 12 weeks after step (a). In some embodiments, step (b) is performed about 9 to about 12 weeks after step (a). In some embodiments, step (b) is performed about 10 to about 12 weeks after step (a). In some embodiments, step (b) is performed about 11 to about 12 weeks after step (a). In some embodiments, step (b) is performed about 4 to about 11 weeks after step (a). In some embodiments, step (b) is performed about 4 to about 10 weeks after step (a). In some embodiments, step (b) is performed about 4 to about 9 weeks after step (a). In some embodiments, step (b) is performed about 4 to about 8 weeks after step (a). In some embodiments, step (b) is performed about 4 to about 7 weeks after step (a). In some embodiments, step (b) is performed about 4 to about 6 weeks after step (a). In some embodiments, step (b) is performed about 4 to about 5 weeks after step (a). In some embodiments, step (b) is performed about 6 to about 8 weeks after step (a). In some embodiments, step (b) is performed about 7 to about 9 weeks after step (a). In some embodiments, step (b) is performed about 8 to about 10 weeks after step (a). In some embodiments, step (b) is performed about 9 to about 11 weeks after step (a).

[0171] In some embodiments of the methods provided herein, step (b) is performed about 4 weeks after step (a). In some embodiments, step (b) is performed about 5 weeks after step (a). In some embodiments, step (b) is performed about 6 weeks after step (a). In some embodiments, step (b) is performed about 7 weeks after step (a). In some embodiments, step (b) is performed about 8 weeks after step (a). In some embodiments, step (b) is performed about 9 weeks after step (a). In some embodiments, step (b) is performed about 10 weeks after step (a). In some embodiments, step (b) is performed about 11 weeks after step (a). In some embodiments, step (b) is performed about 12 weeks after step (a).

[0172] In some embodiments of the methods provided herein, the subject's pathological complete response rate (pCRR) is at least 30%. In some embodiments, the subject's pathological complete response rate (pCRR) is at least 35%. In some embodiments, the subject's pathological complete response rate (pCRR) is at least 40%. In some embodiments, the subject's pathological complete response rate (pCRR) is at least 45%. In some embodiments, the subject's pathological complete response rate (pCRR) is at least 50%. In some embodiments, the subject's pathological complete response rate (pCRR) is at least 55%. In some embodiments, the subject's pathological complete response rate (pCRR) is at least 60%. In some embodiments, the subject's pathological complete response rate (pCRR) is at least 65%. In some embodiments, the subject's pathological complete response rate (pCRR) is at least 70%. In some embodiments, the subject's pathological complete response rate (pCRR) is at least 75%.

[0173] In some embodiments of the methods provided herein, a population of subjects is treated with the method and the pathological complete response rate (pCRR) in the treated population is at least 30%. In some embodiments, a population of subjects is treated with the method and the pathological complete response rate (pCRR) in the treated population is at least 35%. In some embodiments, a population of subjects is treated with the method and the pathological complete response rate (pCRR) in the treated population is at least 40%. In some embodiments, a population of subjects is treated with the method and the pathological complete response rate (pCRR) in the treated population is at least 45%. In some embodiments, a population of subjects is treated with the method and the pathological complete response rate (pCRR) in the treated population is at least 50%. In some embodiments, a population of subjects is treated with the method and the pathological complete response rate (pCRR) in the treated population is at least 55%. In some embodiments, a population of subjects is treated with the method and the pathological complete response rate (pCRR) in the treated population is at least 60%. In some embodiments, a population of subjects is treated with the method and the pathological complete response rate (pCRR) in the treated population is at least 65%. In some embodiments, a population of subjects is treated with the method and the pathological complete response rate (pCRR) in the treated population is at least 70%. In some embodiments, a population of subjects is treated with the method and the pathological complete response rate (pCRR) in the treated population is at least 75%.

[0174] In some embodiments of the methods provided herein, the pathological complete response rate (pCRR) of a population of subjects treated with an ADC is at least 30%. In some embodiments, the pathological complete response rate (pCRR) of a population of subjects treated with an ADC is at least 35%. In some embodiments, the pathological complete response rate (pCRR) of a population of subjects treated with an ADC is at least 40%. In some embodiments, the pathological complete response rate (pCRR) of a population of subjects treated with an ADC is at least 45%. In some embodiments, the pathological complete response rate (pCRR) of a population of subjects treated with an ADC is at least 50%. In some embodiments, the pathological complete response rate (pCRR) of a population of subjects treated with an ADC is at least 55%. In some embodiments, the pathological complete response rate (pCRR) of a population of subjects treated with an ADC is at least 60%. In some embodiments, the pathological complete response rate (pCRR) of a population of subjects treated with an ADC is at least 65%. In some embodiments, the pathological complete response rate (pCRR) in a population of subjects treated with the ADC is at least 70%. In some embodiments, the pathological complete response rate (pCRR) in a population of subjects treated with the ADC is at least 75%.

[0175] In some embodiments of the methods provided herein, the subject's pathological downstaging rate (pDSR) is at least 50%. In some embodiments, the subject's pathological downstaging rate (pDSR) is at least 55%. In some embodiments, the subject's pathological downstaging rate (pDSR) is at least 60%. In some embodiments, the subject's pathological downstaging rate (pDSR) is at least 65%. In some embodiments, the subject's pathological downstaging rate (pDSR) is at least 70%. In some embodiments, the subject's pathological downstaging rate (pDSR) is at least 75%. In some embodiments, the subject's pathological downstaging rate (pDSR) is at least 80%. In some embodiments, the subject's pathological downstaging rate (pDSR) is at least 85%. In some embodiments, the subject's pathological downstaging rate (pDSR) is at least 90%.

[0176] In some embodiments of the methods provided herein, a population of subjects is treated with the method and the pathological downstaging rate (pDSR) in the treated population is at least 50%. In some embodiments, a population of subjects is treated with the method and the pathological downstaging rate (pDSR) in the treated population is at least 55%. In some embodiments, a population of subjects is treated with the method and the pathological downstaging rate (pDSR) in the treated population is at least 60%. In some embodiments, a population of subjects is treated with the method and the pathological downstaging rate (pDSR) in the treated population is at least 65%. In some embodiments, a population of subjects is treated with the method and the pathological downstaging rate (pDSR) in the treated population is at least 70%. In some embodiments, a population of subjects is treated with the method and the pathological downstaging rate (pDSR) in the treated population is at least 75%. In some embodiments, a population of subjects is treated by the method and the pathological downstaging rate (pDSR) in the treated population is at least 80%. In some embodiments, a population of subjects is treated by the method and the pathological downstaging rate (pDSR) in the treated population is at least 85%. In some embodiments, a population of subjects is treated by the method and the pathological downstaging rate (pDSR) in the treated population is at least 90%.

[0177] In some embodiments of the methods provided herein, the pathological downstaging rate (pDSR) of a population of subjects treated with an ADC is at least 50%. In some embodiments, the pathological downstaging rate (pDSR) of a population of subjects treated with an ADC is at least 55%. In some embodiments, the pathological downstaging rate (pDSR) of a population of subjects treated with an ADC is at least 60%. In some embodiments, the pathological downstaging rate (pDSR) of a population of subjects treated with an ADC is at least 65%. In some embodiments, the pathological downstaging rate (pDSR) of a population of subjects treated with an ADC is at least 70%. In some embodiments, the pathological downstaging rate (pDSR) of a population of subjects treated with an ADC is at least 75%.

[0178] In one aspect, a method of treating urothelial carcinoma or bladder cancer in a human subject is provided, comprising administering to the subject multiple cycles of an effective amount of an antibody-drug conjugate (ADC), wherein (a) the ADC comprises an antibody or antigen-binding fragment thereof that binds to 191P4D12 (Nectin-4) conjugated to one or more units of monomethyl auristatin E (MMAE), (b) the subject is ineligible to receive cisplatin therapy (cisplatin-ineligible), and (c) the number of cycles of ADC treatment administered to the cisplatin-ineligible subject is equal to or less than the number of cycles of standard of care (SOC) therapy used to treat cisplatin-eligible subjects with MIUC or MIBC. In some embodiments, an effective amount of the ADC is administered to the subject on days 1 and 8 of every 3-week cycle for a total of three cycles. In some embodiments, the effective amount of the ADC is about 1.25 mg / kg of the subject's body weight. In some embodiments, the effective amount of the ADC is about 1.0 mg / kg of the subject's body weight.

[0179] In one aspect, provided is a method of treating muscle-invasive urothelial carcinoma (MIUC) or muscle-invasive bladder cancer (MIBC) in a human subject, comprising administering to the subject multiple cycles of an effective amount of an antibody-drug conjugate (ADC), wherein (a) the ADC comprises an antibody or antigen-binding fragment thereof that binds to 191P4D12 (Nectin-4) conjugated to one or more units of monomethyl auristatin E (MMAE); (b) the subject is ineligible to receive cisplatin treatment (cisplatin-ineligible); and (c) the number of cycles of ADC treatment administered to the cisplatin-ineligible subject is equal to or less than the number of cycles of standard of care (SOC) therapy used to treat cisplatin-eligible subjects with MIUC or MIBC.

[0180] In some embodiments of the methods provided herein, the SOC therapy for the cisplatin-eligible subject includes cisplatin. In some embodiments, the SOC therapy for the cisplatin-eligible subject includes (i) methotrexate, vinblastine, doxorubicin, and cisplatin (MVAC), or (ii) gemcitabine and cisplatin. In some embodiments, the SOC therapy for the cisplatin-eligible subject includes methotrexate, vinblastine, doxorubicin, and cisplatin (MVAC). In some embodiments, the SOC therapy for the cisplatin-eligible subject includes gemcitabine and cisplatin. In some embodiments, the SOC therapy for the cisplatin-eligible subject includes administering a programmed cell death 1 (PD-1) inhibitor or a programmed cell death-ligand 1 (PD-L1) inhibitor. In some embodiments, SOC therapy for cisplatin-eligible subjects includes i. cisplatin, ii. methotrexate, vinblastine, doxorubicin, and cisplatin (MVAC), iii. gemcitabine and cisplatin, iv. administering a programmed cell death 1 (PD-1) inhibitor, or v. administering a programmed cell death-ligand 1 (PD-L1) inhibitor.

[0181] In some embodiments of the methods provided herein, the number of cycles of ADC therapy administered to the cisplatin-ineligible subject is 1, 2, 3, or 4. In some embodiments, the number of cycles of ADC therapy administered to the cisplatin-ineligible subject is 3 or 4. In some embodiments, the number of cycles of ADC therapy administered to the cisplatin-ineligible subject is 4. In some embodiments, the number of cycles of ADC therapy administered to the cisplatin-ineligible subject is 3. In some embodiments, the number of cycles of ADC therapy administered to the cisplatin-ineligible subject is 2. In some embodiments, the number of cycles of ADC therapy administered to the cisplatin-ineligible subject is 1.

[0182] In some embodiments of the methods provided herein, the method further comprises performing a radical cystectomy and pelvic lymph node dissection (RC+PLND) surgery on the subject. In some embodiments, the RC+PLND is performed about 4 to about 12 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 5 to about 12 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 6 to about 12 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 7 to about 12 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 8 to about 12 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 9 to about 12 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 10 to about 12 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 11 to about 12 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 4 to about 11 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 4 to about 10 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 4 to about 9 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 4 to about 8 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 4 to about 7 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 4 to about 6 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 4 to about 5 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 6 to about 8 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 7 to about 9 weeks after the ADC is administered to the human subject.In some embodiments, the RC+PLND is performed about 8 to about 10 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 9 to about 11 weeks after the ADC is administered to the human subject.

[0183] In some embodiments, the RC+PLND is performed about 4 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 5 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 6 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 7 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 8 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 9 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 10 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 11 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 12 weeks after the ADC is administered to the human subject.

[0184] In one aspect, provided herein are methods of administering neoadjuvant or perioperative therapy for treating muscle-invasive urothelial carcinoma (MIUC) or muscle-invasive bladder cancer (MIBC) in a human subject, the method comprising administering to the subject multiple cycles an effective amount of an antibody-drug conjugate (ADC), wherein (a) the ADC comprises an antibody or antigen-binding fragment thereof that binds to 191P4D12 (Nectin-4) conjugated to one or more units of monomethyl auristatin E (MMAE), (b) the subject is ineligible to receive cisplatin treatment (cisplatin-ineligible), and (c) the subject remains eligible for radical cystectomy and pelvic lymph node dissection (RC+PLND) surgery after multiple cycles of treatment with the ADC. In some embodiments, an effective amount of the ADC is administered to the subject on days 1 and 8 of every 3-week cycle for a total of three cycles. In some embodiments, the effective amount of the ADC is about 1.25 mg / kg of the subject's body weight. In some embodiments, the effective amount of ADC is about 1.0 mg / kg of subject body weight.

[0185] In some embodiments, the method further comprises performing a radical cystectomy and pelvic lymph node dissection (RC+PLND) surgery on the subject. In some embodiments, the ADC is administered as neoadjuvant therapy before surgery. In some embodiments, the ADC is administered before and after surgery. In some embodiments, the ADC is administered as neoadjuvant therapy (i) before surgery, or (ii) before and after surgery. In some embodiments, the RC+PLND is performed about 4 to about 12 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 5 to about 12 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 6 to about 12 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 7 to about 12 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 8 to about 12 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 9 to about 12 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 10 to about 12 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 11 to about 12 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 4 to about 11 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 4 to about 10 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 4 to about 9 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 4 to about 8 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 4 to about 7 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 4 to about 6 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 4 to about 5 weeks after the ADC is administered to the human subject.In some embodiments, RC+PLND is performed about 6 to about 8 weeks after the ADC is administered to the human subject. In some embodiments, RC+PLND is performed about 7 to about 9 weeks after the ADC is administered to the human subject. In some embodiments, RC+PLND is performed about 8 to about 10 weeks after the ADC is administered to the human subject. In some embodiments, RC+PLND is performed about 9 to about 11 weeks after the ADC is administered to the human subject.

[0186] In some embodiments, the RC+PLND is performed about 4 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 5 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 6 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 7 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 8 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 9 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 10 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 11 weeks after the ADC is administered to the human subject. In some embodiments, the RC+PLND is performed about 12 weeks after the ADC is administered to the human subject.

[0187] In some embodiments, the ADC is administered about 1 week after surgery. In some embodiments, the ADC is administered about 2 weeks after surgery. In some embodiments, the ADC is administered about 3 weeks after surgery. In some embodiments, the ADC is administered about 4 weeks after surgery. In some embodiments, the ADC is administered about 5 weeks after surgery. In some embodiments, the ADC is administered about 6 weeks after surgery. In some embodiments, the ADC is administered about 2 months after surgery. In some embodiments, the ADC is administered about 3 months after surgery. In some embodiments, the ADC is administered about 4 months after surgery. In some embodiments, the ADC is administered about 5 months after surgery. In some embodiments, the ADC is administered about 6 months after surgery. In some embodiments, the ADC is administered about 7 months after surgery. In some embodiments, the ADC is administered about 8 months after surgery. In some embodiments, the ADC is administered about 9 months after surgery. In some embodiments, the ADC is administered about 10 months after surgery. In some embodiments, the ADC is administered about 11 months after surgery. In some embodiments, the ADC is administered about 12 months after surgery.

[0188] Standard of care (SOC) therapies for the treatment of muscle-invasive bladder cancer (MIBC) include cisplatin-based neoadjuvant chemotherapy (NAC) (plus definitive local treatment (cystectomy or radiation)) (see, e.g., Advanced Bladder Cancer Meta-analysis, Eur Urol., 2005, 48(2):202-5), accelerated or dose-dense methotrexate, vinblastine, doxorubicin, and cisplatin (MVAC) (see, e.g., Choueiri et al., J Clin Oncol, 2014, 32(18):1889-94; Plimack et al., J Clin Oncol., 2014, 32(18):1895-901), gemcitabine and cisplatin, and programmed cell death 1 (PD-1) inhibitors (see, e.g., Necchi et al., J Clin Oncol., 2018, 36(34):3353-60), and programmed cell death-ligand 1 (PD-L1) inhibitors (see, e.g., Powles et al., 2018, J Clin Oncol., 36(Suppl 15):Abstract 4506). Exemplary measures of treatment efficacy include pathological complete response rate (pCRR), pathological downstaging rate (pDSR), disease-free survival (DFS), recurrence-free survival (EFS), overall survival (OS), progression-free survival (PFS), and / or duration of response (DoR). For example, a 5-year OS of 50% was observed in patients treated with cisplatin-based NAC, compared with an OS of 45% in patients who received definitive local therapy alone (see, e.g., Advanced Bladder Cancer Meta-analysis, Eur Urol., 2005, 48(2):202-5). In patients treated with accelerated or dose-dense MVAC, approximately 30%–40% of patients achieved pCR rates and 50% had pathological downstaging (Choueiri 2014, Plimack 2014).In patients treated with pembrolizumab (a PD-1 inhibitor), a 42% pCR rate and a 65.7% pathological downstaging rate were observed among 50 treated patients, with a pCR rate of 54.3% in patients with PD-L1-positive disease (Necchi 2018). In patients treated with atezolizumab (a PD-L1 inhibitor), pCR was observed in 29% of patients and 40% of patients with PD-L1-positive disease, compared to only 16% in the PD-L1-negative cohort, with a total of 39% of patients downstaging to non-muscle-invasive disease (Powles 2018).

[0189] In one aspect, provided herein is a method of treating muscle-invasive urothelial carcinoma (MIUC) or muscle-invasive bladder cancer (MIBC) in a human subject, the method comprising administering to the subject multiple cycles of an effective amount of an antibody-drug conjugate (ADC), wherein (a) the ADC comprises an antibody or antigen-binding fragment thereof that binds to 191P4D12 (Nectin-4) conjugated to one or more units of monomethyl auristatin E (MMAE); (b) the subject is ineligible to receive cisplatin therapy (cisplatin-ineligible); and (c) the effectiveness of treatment of the subject with the effective amount of the ADC is approximately as effective as treatment observed in cisplatin-eligible patients treated with standard of care (SOC) therapy used to treat cisplatin-eligible subjects with MIUC or MIBC.

[0190] In one aspect, provided herein is a method of treating muscle-invasive urothelial carcinoma (MIUC) or muscle-invasive bladder cancer (MIBC) in a human subject, the method comprising administering to the subject multiple cycles of an effective amount of an antibody-drug conjugate (ADC), wherein (a) the ADC comprises an antibody or antigen-binding fragment thereof that binds to 191P4D12 (Nectin-4) conjugated to one or more units of monomethyl auristatin E (MMAE); (b) the subject is ineligible to receive cisplatin therapy (cisplatin-ineligible); and (c) the efficacy of treatment of the subject with the effective amount of the ADC is the same as the efficacy of treatment observed in cisplatin-eligible patients treated with standard of care (SOC) therapy used to treat cisplatin-eligible subjects with MIUC or MIBC.

[0191] In one aspect, provided herein are methods of treating muscle-invasive urothelial carcinoma (MIUC) or muscle-invasive bladder cancer (MIBC) in a human subject, comprising administering to the subject multiple cycles of an effective amount of an antibody-drug conjugate (ADC), wherein (a) the ADC comprises an antibody or antigen-binding fragment thereof that binds to 191P4D12 (Nectin-4) conjugated to one or more units of monomethyl auristatin E (MMAE), (b) the subject is ineligible to receive cisplatin therapy (cisplatin-ineligible), and (c) the efficacy of treatment of the subject with the effective amount of the ADC is similar to the efficacy of treatment observed in cisplatin-eligible patients treated with standard of care (SOC) therapy used to treat cisplatin-eligible subjects with MIUC or MIBC. In some embodiments, the effective amount of the ADC is administered to the subject on days 1 and 8 of every 3-week cycle for a total of three cycles. In some embodiments, the effective amount of the ADC is about 1.25 mg / kg of the subject's body weight. In some embodiments, the effective amount of ADC is about 1.0 mg / kg of subject body weight.

[0192] In some embodiments, the efficacy of the treatment is at least as effective as the efficacy of the treatment observed in the SOC for cisplatin-eligible subjects. In some embodiments, the measure of efficacy of the treatment is one or more of pathological complete response rate (pCRR), pathological downstaging rate (pDSR), disease-free survival (DFS), recurrence-free survival (EFS), overall survival (OS), progression-free survival (PFS), and duration of response (DoR). In some embodiments, the measure of efficacy of the treatment is pathological complete response rate (pCRR). In some embodiments, the measure of efficacy of the treatment is pathological downstaging rate (pDSR). In some embodiments, the measure of efficacy of the treatment is disease-free survival (DFS). In some embodiments, the measure of efficacy of the treatment is recurrence-free survival (EFS). In some embodiments, the measure of efficacy of the treatment is overall survival (OS). In some embodiments, the measure of efficacy of the treatment is progression-free survival (PFS). In some embodiments, the measure of efficacy of treatment is duration of response (DoR).

[0193] In some embodiments of the methods provided herein, the subject's pathological complete response rate (pCRR) is at least 30%. In some embodiments, the subject's pathological complete response rate (pCRR) is at least 35%. In some embodiments, the subject's pathological complete response rate (pCRR) is at least 40%. In some embodiments, the subject's pathological complete response rate (pCRR) is at least 45%. In some embodiments, the subject's pathological complete response rate (pCRR) is at least 50%. In some embodiments, the subject's pathological complete response rate (pCRR) is at least 55%. In some embodiments, the subject's pathological complete response rate (pCRR) is at least 60%. In some embodiments, the subject's pathological complete response rate (pCRR) is at least 65%. In some embodiments, the subject's pathological complete response rate (pCRR) is at least 70%. In some embodiments, the subject's pathological complete response rate (pCRR) is at least 75%.

[0194] In some embodiments of the methods provided herein, the pathological complete response rate (pCRR) of a population of subjects treated with an ADC is at least 30%. In some embodiments, the pathological complete response rate (pCRR) of a population of subjects treated with an ADC is at least 35%. In some embodiments, the pathological complete response rate (pCRR) of a population of subjects treated with an ADC is at least 40%. In some embodiments, the pathological complete response rate (pCRR) of a population of subjects treated with an ADC is at least 45%. In some embodiments, the pathological complete response rate (pCRR) of a population of subjects treated with an ADC is at least 50%. In some embodiments, the pathological complete response rate (pCRR) of a population of subjects treated with an ADC is at least 55%. In some embodiments, the pathological complete response rate (pCRR) of a population of subjects treated with an ADC is at least 60%. In some embodiments, the pathological complete response rate (pCRR) of a population of subjects treated with an ADC is at least 65%. In some embodiments, the pathological complete response rate (pCRR) in a population of subjects treated with the ADC is at least 70%. In some embodiments, the pathological complete response rate (pCRR) in a population of subjects treated with the ADC is at least 75%.

[0195] In some embodiments, the subject's pathological downstaging rate (pDSR) is at least 50%. In some embodiments, the subject's pathological downstaging rate (pDSR) is at least 55%. In some embodiments, the subject's pathological downstaging rate (pDSR) is at least 60%. In some embodiments, the subject's pathological downstaging rate (pDSR) is at least 65%. In some embodiments, the subject's pathological downstaging rate (pDSR) is at least 70%. In some embodiments, the subject's pathological downstaging rate (pDSR) is at least 75%. In some embodiments, the subject's pathological downstaging rate (pDSR) is at least 80%. In some embodiments, the subject's pathological downstaging rate (pDSR) is at least 85%. In some embodiments, the subject's pathological downstaging rate (pDSR) is at least 90%.

[0196] In some embodiments of the methods provided herein, the pathological downstaging rate (pDSR) of a population of subjects treated with an ADC is at least 50%. In some embodiments, the pathological downstaging rate (pDSR) of a population of subjects treated with an ADC is at least 55%. In some embodiments, the pathological downstaging rate (pDSR) of a population of subjects treated with an ADC is at least 60%. In some embodiments, the pathological downstaging rate (pDSR) of a population of subjects treated with an ADC is at least 65%. In some embodiments, the pathological downstaging rate (pDSR) of a population of subjects treated with an ADC is at least 70%. In some embodiments, the pathological downstaging rate (pDSR) of a population of subjects treated with an ADC is at least 75%.

[0197] In some embodiments of the methods provided herein, the SOC therapy for the cisplatin-eligible subject includes cisplatin. In some embodiments, the SOC therapy for the cisplatin-eligible subject includes (i) methotrexate, vinblastine, doxorubicin, and cisplatin (MVAC), or (ii) gemcitabine and cisplatin. In some embodiments, the SOC therapy for the cisplatin-eligible subject includes methotrexate, vinblastine, doxorubicin, and cisplatin (MVAC). In some embodiments, the SOC therapy for the cisplatin-eligible subject includes gemcitabine and cisplatin. In some embodiments, the SOC therapy for the cisplatin-eligible subject includes a programmed cell death 1 (PD-1) inhibitor or a programmed cell death-ligand 1 (PD-L1) inhibitor. In some embodiments, the SOC therapy for a cisplatin-eligible subject comprises i. cisplatin, ii. methotrexate, vinblastine, doxorubicin, and cisplatin (MVAC), iii. gemcitabine and cisplatin, iv. a programmed cell death 1 (PD-1) inhibitor, or v. a programmed cell death-ligand 1 (PD-L1) inhibitor. In some embodiments, the SOC therapy for a cisplatin-eligible subject comprises a programmed cell death 1 (PD-1) inhibitor. In some embodiments, the SOC therapy for a cisplatin-eligible subject comprises a programmed cell death-ligand 1 (PD-L1) inhibitor. In some embodiments, the PD-1 inhibitor is nivolumab or pembrolizumab. In some embodiments, the PD-1 inhibitor is nivolumab. In some embodiments, the PD-1 inhibitor is pembrolizumab. In some embodiments, the PD-L1 inhibitor is selected from the group consisting of atezolizumab, avelumab, and durvalumab. In some embodiments, the PD-L1 inhibitor is atezolizumab, avelumab, or durvalumab. In some embodiments, the PD-L1 inhibitor is atezolizumab. In some embodiments, the PD-L1 inhibitor is avelumab. In some embodiments, the PD-L1 inhibitor is durvalumab.In some embodiments, (i) the PD-1 inhibitor is nivolumab or pembrolizumab, or (ii) the PD-L1 inhibitor is selected from the group consisting of atezolizumab, avelumab, and durvalumab. In some embodiments of the methods provided herein, the subject has cT2-T4aN0M0 stage MIBC.

[0198] In some embodiments of the methods provided herein, the effective amount of ADC is about 1.25 mg / kg.

[0199] In some embodiments of the methods provided herein, the cancer is urothelial carcinoma or bladder cancer. In some embodiments, the cancer is urothelial carcinoma. In some embodiments, the cancer is bladder cancer. In some embodiments, the cancer is muscle-invasive urothelial carcinoma (MIUC). In some embodiments, the cancer is muscle-invasive bladder cancer (MIBC).

[0200] 5.2.1.1 Cisplatin-ineligible patients For the methods provided herein, including but not limited to the method in the preceding paragraph, a variety of conditions can be used to determine cisplatin ineligibility of a human subject.

[0201] In one embodiment, the criteria for determining cisplatin ineligibility include or consist of a GFR less than 60 mL / min but greater than or equal to 30 mL / min, where GFR is measured by the Cockcroft-Gault formula, Modification of Diet in Renal Disease (MDRD), or a 24-hour urine collection. In some embodiments, the criteria for determining cisplatin ineligibility include or consist of an ECOG performance status score of 2. In some embodiments, the criteria for determining cisplatin ineligibility include or consist of hearing loss of grade 2 or greater on the NCI CTCAE version 4.03. In some embodiments, the criteria for determining cisplatin ineligibility include or consist of NYHA class III heart failure. In some embodiments, a subject is considered cisplatin-ineligible if they meet one or more of the following criteria: (a) a GFR of less than 60 mL / min but greater than or equal to 30 mL / min, where GFR is measured by the Cockcroft-Gault formula, Modification of Diet in Renal Disease (MDRD), or a 24-hour urine collection; (b) an ECOG performance status of 2; (c) hearing loss of grade 2 or greater per NCI CTCAE version 4.03; and (d) NYHA class III heart failure. In some embodiments, the criteria for determining cisplatin ineligibility include or consist of a GFR of less than 60 mL / min but greater than or equal to 30 mL / min, where GFR is measured by the Cockcroft-Gault formula, Modification of Diet in Renal Disease (MDRD), or a 24-hour urine collection, and an ECOG performance status score of 2.In some embodiments, the criteria for determining cisplatin ineligibility include or consist of a GFR less than 60 mL / min but greater than or equal to 30 mL / min, where the GFR is measured by the Cockcroft-Gault formula, Modification of Diet in Renal Disease (MDRD), or a 24-hour urine collection, and hearing loss of Grade 2 or greater per NCI CTCAE version 4.03. In some embodiments, the criteria for determining cisplatin ineligibility include or consist of a GFR less than 60 mL / min but greater than or equal to 30 mL / min, where the GFR is measured by the Cockcroft-Gault formula, Modification of Diet in Renal Disease (MDRD), or a 24-hour urine collection, and NYHA Class III heart failure. In some embodiments, the criteria for determining cisplatin ineligibility include or consist of an ECOG performance status score of 2 and hearing loss of Grade 2 or greater per NCI CTCAE version 4.03. In some embodiments, the criteria for determining cisplatin ineligibility include or consist of an ECOG performance status score of 2 and NYHA class III heart failure. In some embodiments, the criteria for determining cisplatin ineligibility include or consist of hearing loss of grade 2 or greater per NCI CTCAE version 4.03 and NYHA class III heart failure. In some embodiments, a human subject for the methods provided herein can have any three of a GFR of less than 60 mL / min but greater than or equal to 30 mL / min (wherein GFR is measured by the Cockcroft-Gault formula, Modification of Diet in Renal Disease formula (MDRD), or a 24-hour urine collection), an ECOG performance status of 2, hearing loss of grade 2 or greater per NCI CTCAE version 4.03, and NYHA class III heart failure.In some embodiments, a subject is considered cisplatin-ineligible if they meet one or more of the following criteria: (a) a GFR of less than 60 mL / min but greater than or equal to 30 mL / min (wherein GFR is measured by the Cockcroft-Gault formula, Modification of Diet in Renal Disease (MDRD), or a 24-hour urine collection), (b) an ECOG performance status score of 2, (c) hearing loss of grade 2 or greater per NCI CTCAE version 4.03, and (d) NYHA class III heart failure. In some embodiments, a human subject for the methods provided herein may have one or more of a GFR of less than 60 mL / min but greater than or equal to 30 mL / min (wherein GFR is measured by the Cockcroft-Gault formula, Modification of Diet in Renal Disease (MDRD), or a 24-hour urine collection), an ECOG performance status score of 2, hearing loss of grade 2 or greater per NCI CTCAE version 4.03, and NYHA class III heart failure.

[0202] In other embodiments, the criteria for determining cisplatin ineligibility include or consist of an ECOG performance status score of 2. In some embodiments, the criteria for determining cisplatin ineligibility include or consist of renal impairment. In certain embodiments, the criteria for determining cisplatin ineligibility include or consist of hearing loss of Grade 2 or greater. In one embodiment, the criteria for determining cisplatin ineligibility include or consist of an ECOG performance status score of 2 and renal impairment. In some embodiments, the criteria for determining cisplatin ineligibility include or consist of an ECOG performance status score of 2 and hearing loss of Grade 2 or greater. In further embodiments, the criteria for determining cisplatin ineligibility include or consist of renal impairment and hearing loss of Grade 2 or greater. In yet other embodiments, the criteria for determining cisplatin ineligibility include or consist of an ECOG performance status score of 2, renal impairment, and hearing loss of Grade 2 or greater. In some embodiments, the conditions for determining cisplatin ineligibility include or consist of any one of an ECOG performance status score of 2, renal impairment, and hearing loss of grade 2 or greater. In some embodiments, the conditions for determining cisplatin ineligibility include or consist of any two of an ECOG performance status score of 2, renal impairment, and hearing loss of grade 2 or greater, in any combination or permutation. In some embodiments, the conditions for determining cisplatin ineligibility include or consist of all three of an ECOG performance status score of 2, renal impairment, and hearing loss of grade 2 or greater.

[0203] Renal dysfunction can be determined by various means known and available in the art. Various embodiments for determining renal dysfunction in a human subject are provided herein for the methods provided herein, including but not limited to the method in the preceding paragraph. In one embodiment, renal dysfunction is determined by a creatinine clearance (CrCl) of less than 60 mL / min. In some embodiments, renal dysfunction is determined by a CrCl of less than 60 mL / min but greater than or equal to 30 mL / min. In certain embodiments, renal dysfunction is determined by a CrCl of less than 30 mL / min but greater than or equal to 15 mL / min. In some embodiments of the methods provided in this paragraph, CrCl is measured by a 24-hour urine collection. In other embodiments of the methods provided in this paragraph, CrCl is estimated by the Cockcroft-Gault criteria.

[0204] Thus, for the methods provided herein, including but not limited to the method in the preceding paragraph, several specific criteria based on creatinine clearance for determining cisplatin ineligibility of a human subject can be used. In some embodiments, the criteria for determining cisplatin ineligibility include or consist of a CrCl of less than 60 mL / min. In one embodiment, the criteria for determining cisplatin ineligibility include or consist of an ECOG performance status score of 2 and a CrCl of less than 60 mL / min. In a further embodiment, the criteria for determining cisplatin ineligibility include or consist of a CrCl of less than 60 mL / min and hearing loss of grade 2 or greater. In yet other embodiments, the criteria for determining cisplatin ineligibility include or consist of an ECOG performance status score of 2, a CrCl of less than 60 mL / min, and hearing loss of grade 2 or greater. In some embodiments, the criteria for determining cisplatin ineligibility include or consist of any one of an ECOG performance status score of 2, a CrCl of less than 60 mL / min, and hearing loss of grade 2 or greater. In some embodiments, the criteria for determining cisplatin ineligibility include or consist of any two of the following, in any combination or permutation: an ECOG performance status score of 2, a CrCl of less than 60 mL / min, and a hearing loss of Grade 2 or greater. In some embodiments, the criteria for determining cisplatin ineligibility include or consist of all three of the following: an ECOG performance status score of 2, a CrCl of less than 60 mL / min, and a hearing loss of Grade 2 or greater. In some embodiments of the methods provided in this paragraph, CrCl is measured by a 24-hour urine collection. In other embodiments of the methods provided in this paragraph, CrCl is estimated by the Cockcroft-Gault criteria.

[0205] Alternatively, other specific criteria based on creatinine clearance for determining cisplatin ineligibility of a human subject can be used for the methods provided herein, including, but not limited to, the method in the preceding paragraph. In some embodiments, the criteria for determining cisplatin ineligibility include or consist of a CrCl of less than 60 mL / min but greater than or equal to 30 mL / min. In one embodiment, the criteria for determining cisplatin ineligibility include or consist of an ECOG performance status score of 2 and a CrCl of less than 60 mL / min but greater than or equal to 30 mL / min. In a further embodiment, the criteria for determining cisplatin ineligibility include or consist of a CrCl of less than 60 mL / min but greater than or equal to 30 mL / min and hearing loss of grade 2 or greater. In yet other embodiments, the criteria for determining cisplatin ineligibility include or consist of an ECOG performance status score of 2, a CrCl of less than 60 mL / min but greater than or equal to 30 mL / min, and hearing loss of grade 2 or greater. In some embodiments, the criteria for determining cisplatin ineligibility include or consist of any one of an ECOG performance status score of 2, a CrCl of less than 60 mL / min but greater than or equal to 30 mL / min, and a hearing loss of Grade 2 or greater. In some embodiments, the criteria for determining cisplatin ineligibility include or consist of any two of an ECOG performance status score of 2, a CrCl of less than 60 mL / min but greater than or equal to 30 mL / min, and a hearing loss of Grade 2 or greater, in any combination or permutation. In some embodiments, the criteria for determining cisplatin ineligibility include or consist of all three of an ECOG performance status score of 2, a CrCl of less than 60 mL / min but greater than or equal to 30 mL / min, and a hearing loss of Grade 2 or greater. In some embodiments of the methods provided in this paragraph, CrCl is measured by a 24-hour urine collection. In other embodiments of the methods provided in this paragraph, CrCl is estimated by the Cockcroft-Gault criteria.

[0206] Similarly, for the methods provided herein, including but not limited to the method in the preceding paragraph, more specific criteria based on creatinine clearance for determining cisplatin ineligibility of a human subject can be used. In some embodiments, the criteria for determining cisplatin ineligibility include or consist of a CrCl of less than 30 mL / min but greater than or equal to 15 mL / min. In one embodiment, the criteria for determining cisplatin ineligibility include or consist of an ECOG performance status score of 2 and a CrCl of less than 30 mL / min but greater than or equal to 15 mL / min. In a further embodiment, the criteria for determining cisplatin ineligibility include or consist of a CrCl of less than 30 mL / min but greater than or equal to 15 mL / min and hearing loss of grade 2 or greater. In yet other embodiments, the criteria for determining cisplatin ineligibility include or consist of an ECOG performance status score of 2, a CrCl of less than 30 mL / min but greater than or equal to 15 mL / min, and hearing loss of grade 2 or greater. In some embodiments, the criteria for determining cisplatin ineligibility include or consist of any one of an ECOG performance status score of 2, a CrCl of less than 30 mL / min but greater than or equal to 15 mL / min, and a hearing loss of Grade 2 or greater. In some embodiments, the criteria for determining cisplatin ineligibility include or consist of any two of an ECOG performance status score of 2, a CrCl of less than 30 mL / min but greater than or equal to 15 mL / min, and a hearing loss of Grade 2 or greater, in any combination or permutation. In some embodiments, the criteria for determining cisplatin ineligibility include or consist of all three of an ECOG performance status score of 2, a CrCl of less than 30 mL / min but greater than or equal to 15 mL / min, and a hearing loss of Grade 2 or greater. In some embodiments of the methods provided in this paragraph, CrCl is measured by a 24-hour urine collection. In other embodiments of the methods provided in this paragraph, CrCl is estimated by the Cockcroft-Gault criteria.

[0207] 5.2.1.2 Additional Patient Demographics In addition, human subjects with a variety of other conditions may be used with the methods provided herein. In one embodiment, a human subject for the methods provided herein may have histologically confirmed MIBC with a predominant urothelial histology of greater than 50% (formerly known as transitional cell carcinoma). In another embodiment, a human subject has cT2-T4aN0M0 stage MIBC. In another embodiment, a human subject for the methods provided herein may have clinical stage cT2-T4aN0M0 as determined by TURBT and by CT with IV contrast of the chest and CT urogram of the abdomen and pelvis within 90 days prior to the first administration of treatment. In another embodiment, a human subject for the methods provided herein may have mixed cell types, with a predominant urothelial carcinoma (>50%). In another embodiment, a human subject for the methods provided herein may have an ECOG performance status of 0, 1, or 2. In another embodiment, a human subject for the methods provided herein may have an expected life expectancy of 3 months or greater. In other embodiments, a human subject for the methods provided herein may be deemed eligible for RC+PLND by his / her urologist and / or oncologist. In some embodiments, a human subject for the methods provided herein may have any two of the following: histologically confirmed MIBC (formerly known as transitional cell carcinoma) with a predominant urothelial histology of greater than 50%; clinical stage cT2-T4aN0M0 as determined by TURBT and by CT with IV contrast of the chest and CT urogram of the abdomen and pelvis within 90 days prior to the first dose of treatment; mixed cellularity with predominant (>50%) urothelial carcinoma; ECOG performance status of 0, 1, or 2; predicted life expectancy of 3 months or greater; and deemed eligible for RC+PLND by his / her urologist and / or oncologist.In some embodiments, a human subject for the methods provided herein may have any three of the following: histologically confirmed MIBC (formerly known as transitional cell carcinoma) with greater than 50% predominant urothelial histology; clinical stage cT2-T4aN0M0 as determined by TURBT and by CT with IV contrast of the chest and CT urogram of the abdomen and pelvis within 90 days prior to the first dose of treatment; mixed cellularity with predominant (>50%) urothelial carcinoma; ECOG performance status of 0, 1, or 2; predicted life expectancy of 3 months or greater; and is deemed eligible for RC+PLND by his / her urologist and / or oncologist. In some embodiments, a human subject for the methods provided herein may have any four of the following: histologically confirmed MIBC (formerly known as transitional cell carcinoma) with greater than 50% predominant urothelial histology; clinical stage cT2-T4aN0M0 as determined by TURBT and by CT with IV contrast of the chest and CT urogram of the abdomen and pelvis within 90 days prior to the first dose of treatment; mixed cellularity with predominant (>50%) urothelial carcinoma; ECOG performance status of 0, 1, or 2; predicted life expectancy of 3 months or greater; and is deemed eligible for RC+PLND by his / her urologist and / or oncologist. In some embodiments, a human subject for the methods provided herein may have any five of the following: histologically confirmed MIBC (formerly known as transitional cell carcinoma) with greater than 50% predominant urothelial histology; clinical stage cT2-T4aN0M0 as determined by TURBT and by CT with IV contrast of the chest and CT urogram of the abdomen and pelvis within 90 days prior to the first dose of treatment; mixed cellularity with predominant (>50%) urothelial carcinoma; ECOG performance status of 0, 1, or 2; expected life expectancy of 3 months or greater; and is deemed eligible for RC+PLND by his / her urologist and / or oncologist.In some embodiments, a human subject for the methods provided herein may have all five of the following: histologically confirmed MIBC (formerly known as transitional cell carcinoma) with greater than 50% predominant urothelial histology; clinical stage cT2-T4aN0M0 as determined by TURBT and by CT with IV contrast of the chest and CT urogram of the abdomen and pelvis within 90 days prior to the first dose of treatment; mixed cellularity with predominant (>50%) urothelial carcinoma; ECOG performance status of 0, 1, or 2; predicted life expectancy of 3 months or greater; and is deemed eligible for RC+PLND by his / her urologist and / or oncologist. In some embodiments, a human subject for the methods provided herein may have one or more of the following: histologically confirmed MIBC (formerly known as transitional cell carcinoma) with greater than 50% predominant urothelial histology; clinical stage cT2-T4aN0M0 as determined by TURBT and by CT with IV contrast of the chest and CT urogram of the abdomen and pelvis within 90 days prior to the first dose of treatment; mixed cellularity with predominant (>50%) urothelial carcinoma; ECOG performance status of 0, 1, or 2; expected life expectancy of 3 months or greater; and is deemed eligible for RC+PLND by his / her urologist and / or oncologist.

[0208] In some embodiments, human subjects for the methods provided herein may have previously received intravesical bacillus calmette-guerin (BCG) or intravesical chemotherapy for NMIBC.

[0209] In further embodiments of the methods provided herein, including the method of the preceding paragraph, the human subject for whom the methods provided herein can be used is a human subject with a variety of other conditions. In one embodiment, the human subject for whom the methods provided herein can be used also has an absolute neutrophil count (ANC) of 1500 / μL or greater. In another embodiment, the human subject for whom the methods provided herein can be used also has a platelet count of 100,000 / μL or greater. In another embodiment, the human subject for whom the methods provided herein can be used also has a hemoglobin of 9.0 g / dL or greater or 5.6 mmol / L or greater. In another embodiment, the human subject for whom the methods provided herein can be used also has a CrCl of 1.5 times the upper limit of normal (ULN) or less. In another embodiment, the human subject for whom the methods provided herein can be used also has a creatinine of 1.5 times the upper limit of normal (ULN) or less. In another embodiment, the human subject for whom the methods provided herein can be used also has a CrCl of 30 mL / min or greater. In other embodiments, a human subject for whom the methods provided herein can be used also has (i) a serum total bilirubin level of 1.5 times the upper limit of normal (ULN) or less, (ii) a direct bilirubin level of ULN or less for patients with total bilirubin levels greater than 1.5 times the ULN, or (iii) a serum total bilirubin level of 3 times the ULN for patients with Gilbert's disease. In other embodiments, a human subject for whom the methods provided herein can be used also has an AST (SGOT) and ALT (SGPT) level of 3 times the ULN or less. In other embodiments, a human subject for whom the methods provided herein can be used also has an INR or PT level of 1.5 times the upper limit of normal (ULN) or less. In other embodiments, a human subject for whom the methods provided herein can be used also has an aPTT or PTT level of 1.5 times the upper limit of normal (ULN).In some embodiments, a human subject in whom the methods provided herein can be used also has any two of the following: an absolute neutrophil count (ANC) of 1500 / μL or greater; a platelet count of 100,000 / μL or greater; a hemoglobin of 9.0 g / dL or greater or 5.6 mmol / L or greater; a CrCl of 1.5 times the upper limit of normal (ULN); a creatinine of 1.5 times the upper limit of normal (ULN); a CrCl of 30 mL / min or greater; (i) serum total blood cholesterol (e.g., creatinine ... (ii) direct bilirubin ≤ 1.5 times the upper limit of normal (ULN); (ii) for patients with total bilirubin levels greater than 1.5 times the ULN, direct bilirubin ≤ ULN; or (iii) for patients with Gilbert's disease, serum total bilirubin ≤ 3 times the ULN; AST (SGOT) and ALT (SGPT) ≤ 3 times the ULN; INR or PT ≤ 1.5 times the upper limit of normal (ULN); and aPTT or PTT ≤ 1.5 times the upper limit of normal (ULN). In some embodiments, a human subject for whom the methods provided herein can be used also has any three of the following: an absolute neutrophil count (ANC) of 1500 / μL or greater; a platelet count of 100,000 / μL or greater; a hemoglobin of 9.0 g / dL or greater than 5.6 mmol / L; a CrCl of 1.5 times the upper limit of normal (ULN); a creatinine of 1.5 times the upper limit of normal (ULN); a CrCl of 30 mL / min or greater; (i) a serum total blood glucose level of 1.5 times the upper limit of normal (ULN); (ii) direct bilirubin ≤ 1.5 times the upper limit of normal (ULN); (ii) for patients with total bilirubin levels greater than 1.5 times the ULN, direct bilirubin ≤ ULN; or (iii) for patients with Gilbert's disease, serum total bilirubin ≤ 3 times the ULN; AST (SGOT) and ALT (SGPT) ≤ 3 times the ULN; INR or PT ≤ 1.5 times the upper limit of normal (ULN); and aPTT or PTT ≤ 1.5 times the upper limit of normal (ULN).In some embodiments, a human subject for whom the methods provided herein can be used also has any four of the following: an absolute neutrophil count (ANC) of 1500 / μL or greater; a platelet count of 100,000 / μL or greater; a hemoglobin of 9.0 g / dL or greater than 5.6 mmol / L; a CrCl of 1.5 times the upper limit of normal (ULN); a creatinine of 1.5 times the upper limit of normal (ULN); a CrCl of 30 mL / min or greater; (i) a serum total blood glucose level of 1.5 times the upper limit of normal (ULN); (ii) direct bilirubin ≤ 1.5 times the upper limit of normal (ULN); (ii) for patients with total bilirubin levels greater than 1.5 times the ULN, direct bilirubin ≤ ULN; or (iii) for patients with Gilbert's disease, serum total bilirubin ≤ 3 times the ULN; AST (SGOT) and ALT (SGPT) ≤ 3 times the ULN; INR or PT ≤ 1.5 times the upper limit of normal (ULN); and aPTT or PTT ≤ 1.5 times the upper limit of normal (ULN). In some embodiments, a human subject for whom the methods provided herein can be used also has any five of the following: an absolute neutrophil count (ANC) of 1500 / μL or greater; a platelet count of 100,000 / μL or greater; a hemoglobin of 9.0 g / dL or greater or 5.6 mmol / L or greater; a CrCl of 1.5 times the upper limit of normal (ULN); a creatinine of 1.5 times the upper limit of normal (ULN); a CrCl of 30 mL / min or greater; (i) serum total blood cholesterol (e.g., creatinine); (ii) direct bilirubin ≤ 1.5 times the upper limit of normal (ULN); (ii) for patients with total bilirubin levels greater than 1.5 times the ULN, direct bilirubin ≤ ULN; or (iii) for patients with Gilbert's disease, serum total bilirubin ≤ 3 times the ULN; AST (SGOT) and ALT (SGPT) ≤ 3 times the ULN; INR or PT ≤ 1.5 times the upper limit of normal (ULN); and aPTT or PTT ≤ 1.5 times the upper limit of normal (ULN).In some embodiments, a human subject for whom the methods provided herein can be used also has any six of the following: an absolute neutrophil count (ANC) of 1500 / μL or greater; a platelet count of 100,000 / μL or greater; a hemoglobin of 9.0 g / dL or greater or 5.6 mmol / L or greater; a CrCl of 1.5 times the upper limit of normal (ULN) or less; a creatinine of 1.5 times the upper limit of normal (ULN) or less; a CrCl of 30 mL / min or greater; (i) serum total blood cholesterol (e.g., creatinine ... (ii) direct bilirubin ≤ 1.5 times the upper limit of normal (ULN); (ii) for patients with total bilirubin levels greater than 1.5 times the ULN, direct bilirubin ≤ ULN; or (iii) for patients with Gilbert's disease, serum total bilirubin ≤ 3 times the ULN; AST (SGOT) and ALT (SGPT) ≤ 3 times the ULN; INR or PT ≤ 1.5 times the upper limit of normal (ULN); and aPTT or PTT ≤ 1.5 times the upper limit of normal (ULN). In some embodiments, a human subject for whom the methods provided herein can be used also has any seven of the following: an absolute neutrophil count (ANC) of 1500 / μL or greater; a platelet count of 100,000 / μL or greater; a hemoglobin of 9.0 g / dL or greater or 5.6 mmol / L or greater; a CrCl of 1.5 times the upper limit of normal (ULN) or less; a creatinine of 1.5 times the upper limit of normal (ULN) or less; a CrCl of 30 mL / min or greater; (i) serum total blood cholesterol (e.g., creatinine ... (ii) direct bilirubin ≤ 1.5 times the upper limit of normal (ULN); (ii) for patients with total bilirubin levels greater than 1.5 times the ULN, direct bilirubin ≤ ULN; or (iii) for patients with Gilbert's disease, serum total bilirubin ≤ 3 times the ULN; AST (SGOT) and ALT (SGPT) ≤ 3 times the ULN; INR or PT ≤ 1.5 times the upper limit of normal (ULN); and aPTT or PTT ≤ 1.5 times the upper limit of normal (ULN).In some embodiments, a human subject for whom the methods provided herein can be used also has any eight of the following: an absolute neutrophil count (ANC) of 1500 / μL or greater; a platelet count of 100,000 / μL or greater; a hemoglobin of 9.0 g / dL or greater or 5.6 mmol / L or greater; a CrCl of 1.5 times the upper limit of normal (ULN); a creatinine of 1.5 times the upper limit of normal (ULN); a CrCl of 30 mL / min or greater; (i) serum total blood cholesterol (e.g., creatinine); (ii) direct bilirubin ≤ 1.5 times the upper limit of normal (ULN); (ii) for patients with total bilirubin levels greater than 1.5 times the ULN, direct bilirubin ≤ ULN; or (iii) for patients with Gilbert's disease, serum total bilirubin ≤ 3 times the ULN; AST (SGOT) and ALT (SGPT) ≤ 3 times the ULN; INR or PT ≤ 1.5 times the upper limit of normal (ULN); and aPTT or PTT ≤ 1.5 times the upper limit of normal (ULN). In some embodiments, a human subject in whom the methods provided herein can be used also has all nine of the following: absolute neutrophil count (ANC) of 1500 / μL or greater; platelet count of 100,000 / μL or greater; hemoglobin of 9.0 g / dL or greater or 5.6 mmol / L or greater; CrCl of 1.5 times the upper limit of normal (ULN); creatinine of 1.5 times the upper limit of normal (ULN); CrCl of 30 mL / min or greater; (i) serum total bilirubin (ii) for patients with total bilirubin levels greater than 1.5 times the ULN, direct bilirubin is less than or equal to the ULN; or (iii) for patients with Gilbert's disease, serum total bilirubin is less than or equal to 3 times the ULN; AST (SGOT) and ALT (SGPT) are less than or equal to 3 times the ULN; INR or PT is less than or equal to 1.5 times the upper limit of normal (ULN); and aPTT or PTT is less than or equal to 1.5 times the upper limit of normal (ULN).In some embodiments, a human subject in whom the methods provided herein can be used also has any one or more of the following: an absolute neutrophil count (ANC) of 1500 / μL or greater; a platelet count of 100,000 / μL or greater; a hemoglobin of 9.0 g / dL or greater or 5.6 mmol / L or greater; a CrCl of 1.5 times the upper limit of normal (ULN) or less; a creatinine of 1.5 times the upper limit of normal (ULN) or less; a CrCl of 30 mL / min or greater; (i) serum total blood cholesterol (e.g., cholesterol deficiency syndrome, urinary tract infection ... (ii) direct bilirubin below 1.5 times the upper limit of normal (ULN), (ii) for patients with total bilirubin levels greater than 1.5 times the ULN, direct bilirubin below ULN, or (iii) for patients with Gilbert's disease, serum total bilirubin below 3 times the ULN; AST (SGOT) and ALT (SGPT) below 3 times the ULN; INR or PT below 1.5 times the upper limit of normal (ULN); and aPTT or PTT below 1.5 times the upper limit of normal (ULN).

[0210] In other embodiments of the methods provided herein, including the method of the preceding paragraph, the human subject on which the methods provided herein can be used is a human subject who does not have a specific condition. In one embodiment, the human subject for the methods provided herein has not received previous systemic treatment, chemoradiotherapy, or radiotherapy for MIBC. In one embodiment, the human subject for the methods provided herein has not received any previous treatment with an immune checkpoint inhibitor (CPI) (e.g., a PD-1 inhibitor, a PD-L1 inhibitor, or a PD-L2 inhibitor, such as atezolizumab, pembrolizumab, nivolumab, durvalumab, or avelumab). In one embodiment, the human subject for the methods provided herein has not received any previous treatment with an agent directed against another stimulatory or co-inhibitory T cell receptor (e.g., a CD137 agonist, a CTLA-4 inhibitor, or an OX-40 agonist). In one embodiment, the human subject for the methods provided herein has no evidence of nodal disease on imaging. In one embodiment, a human subject for the methods provided herein has no evidence of metastatic disease on imaging. In one embodiment, a human subject for the methods provided herein has not undergone a partial cystectomy of the bladder to remove any NMIBC or MIBC. In one embodiment, a human subject for the methods provided herein has no ongoing sensory or motor neuropathy grade 2 or higher. In one embodiment, a human subject for the methods provided herein has no conditions requiring high-dose steroids (>10 mg / day of prednisone or equivalent) or other immunosuppressants. In one embodiment, a human subject for the methods provided herein has not received prior treatment with enfortumab vedotin or other MMAE-based ADCs for urothelial carcinoma. In one embodiment, a human subject for the methods provided herein has no history of another invasive malignancy within three years prior to the first dose of enfortumab vedotin (EV) or evidence of residual disease from a previously diagnosed malignancy. In one embodiment, a human subject for the methods provided herein.In one embodiment, a human subject for the methods provided herein is not currently receiving systemic antibacterial treatment for an active infection (i.e., viral, bacterial, or fungal) at the time of the first dose of enfortumab vedotin. In one embodiment, a human subject for the methods provided herein is not positive for hepatitis B surface antigen and / or anti-hepatitis B core antibody. In one embodiment, a human subject for the methods provided herein does not have an active hepatitis C infection or a known HIV infection. In one embodiment, a human subject for the methods provided herein does not have active tuberculosis. In one embodiment, a human subject for the methods provided herein does not have a documented history of a cerebrovascular event consistent with NYHA class IV (stroke or transient ischemic attack), unstable angina, myocardial infarction, or cardiac symptom (including congestive heart failure) within 6 months prior to the first dose of enfortumab vedotin. In one embodiment, a human subject for the methods provided herein does not have active keratitis or a corneal ulcer. In one embodiment, a human subject for the methods provided herein does not have an active autoimmune disease requiring systemic treatment (i.e., use of disease-modifying agents, corticosteroids, or immunosuppressants) within the past two years. In one embodiment, a human subject for the methods provided herein does not have a history of idiopathic pulmonary fibrosis, organizing pneumonia, drug-induced pneumonia, idiopathic pneumonia, or evidence of active pneumonia on a screening chest CT scan. In one embodiment, a human subject for the methods provided herein does not have a previous allogeneic stem cell or solid organ transplant. In one embodiment, a human subject for the methods provided herein has not received a live-attenuated vaccine within 30 days prior to the first dose of test drug. In one embodiment, a human subject for the methods provided herein does not have uncontrolled diabetes, defined as HbA1c ≥ 8% or HbA1c > 7%-8% with associated diabetic symptoms (polyuria or polydipsia) not otherwise described.

[0211] In all of the methods provided herein, and specifically in the methods described in the preceding paragraph, ADCs that may be used are described in Sections 3, 5.2, 5.3, 5.4, 5.5, and 6; patient selection for treatment is described herein and exemplified in this section (Section 5.2), as well as Sections 3 and 6; dosing regimens and pharmaceutical compositions for administering therapeutic agents are described in this section (Section 5.2), in Sections 5.4, 5.7, and 6 below; and biomarkers that may be used to identify therapeutic agents, select patients, determine the outcome of these methods, and / or serve as criteria in any manner for these methods are described herein and in this section (Section 5.2). Biomarkers are exemplified in Section 5.2, including Section 5.1 and 5.2.2, and Section 6; biomarkers can be determined as described in Section 5.8 or known in the art; treatment outcomes for the methods provided herein are described in this section (Section 5.2, including Section 5.2.1.4) and Sections 3 and 6; additional treatment outcomes for the methods provided herein can be improvement in biomarkers described herein, e.g., biomarkers described and exemplified in this section (Section 5.2, including Section 5.2.2), and Sections 3 and 6; combination therapies including ADCs and other therapeutic agents are described in this section (Section 5.2) and Section 5.5. Accordingly, one of skill in the art will understand that the methods provided herein include all permutations and combinations of patients, therapeutic agents, dosing regimens, biomarkers, and treatment outcomes as described above and below.

[0212] In certain embodiments, the methods provided herein are used to treat a subject with a urothelial carcinoma that expresses 191P4D12 RNA, expresses 191P4D12 protein, or expresses both 191P4D12 RNA and 191P4D12 protein. In one embodiment, the methods provided herein are used to treat a subject with a urothelial carcinoma that expresses 191P4D12 RNA or expresses 191P4D12 protein.

[0213] In certain embodiments, the methods provided herein are used to treat a subject with a urothelial carcinoma that expresses 191P4D12 RNA, expresses 191P4D12 protein, or expresses both 191P4D12 RNA and 191P4D12 protein. In one embodiment, the methods provided herein are used to treat a subject with a urothelial carcinoma that expresses 191P4D12 RNA or expresses 191P4D12 protein.

[0214] In some embodiments, 191P4D12 RNA expression in cancer is determined by polynucleotide hybridization, sequencing (assessing relative abundance of sequences), and / or PCR (including RT-PCR). In some embodiments, 191P4D12 protein expression in cancer is determined by IHC, analysis by fluorescence-activated cell sorting (FACS), and / or Western blotting. In some embodiments, 191P4D12 protein expression in cancer is determined by more than one method. In some embodiments, 191P4D12 protein expression in cancer is determined by two methods: IHC.

[0215] In some embodiments, subjects that can be treated with the methods provided herein have certain phenotypic or genotypic characteristics, hi some embodiments, subjects have any permutation or combination of the phenotypic or genotypic characteristics described herein.

[0216] In some embodiments, phenotypic or genotypic characteristics are determined histologically, cytologically, or both histologically and cytologically. In some embodiments of the methods provided herein, histological and / or cytological determination of phenotypic and / or genotypic characteristics is performed based on recently analyzed tissue as described in the American Society of Clinical Oncology / College of American Pathologists (ASCO / CAP) guidelines, the entire contents of which are incorporated herein by reference. In some embodiments, phenotypic or genotypic characteristics are determined by sequencing, including next-generation sequencing (e.g., NGS from Illumina, Inc.), DNA hybridization, and / or RNA hybridization.

[0217] 5.2.1.3 Optional Exclusion of Patients with Prior Checkpoint Inhibitor (CPI) Therapy In various aspects or embodiments of the methods provided herein, including the methods provided in this section (Section 5.2), e.g., the methods provided in this paragraph and the preceding paragraph, the methods can refer to a subject that has not received any prior treatment with an immune checkpoint inhibitor (e.g., a human subject for the methods provided herein has not received any prior treatment with an immune checkpoint inhibitor (CPI) (e.g., a PD-1 inhibitor, PD-L1 inhibitor, or PD-L2 inhibitor such as atezolizumab, pembrolizumab, nivolumab, durvalumab, or avelumab)).

[0218] As used herein, the term "immune checkpoint inhibitor" or "checkpoint inhibitor" refers to a molecule that reduces, inhibits, antagonizes, or modulates one or more checkpoint proteins, either in whole or in part. Numerous checkpoint proteins are known, including CTLA-4 and its ligands CD80 and CD86; and PD-1 and its ligands PD-L1 and PD-L2 (Pardoll, Nature Reviews Cancer, 2012, 12, 252-264). Other exemplary checkpoint proteins include LAG-3, B7, TIM3 (HAVCR2), OX40 (CD134), GITR, CD137, CD40, VTCN1, IDO1, CD276, PVRIG, TIGIT, CD25 (IL2RA), IFNAR2, IFNAR1, CSF1R, VSIR (VISTA), or HLA. These proteins appear to be involved in costimulatory or inhibitory interactions in T cell responses. Immune checkpoint proteins appear to regulate and maintain self-tolerance as well as the duration and magnitude of physiological immune responses. Immune checkpoint inhibitors include or are derived from antibodies.

[0219] In certain embodiments, the checkpoint inhibitor can be an inhibitor or activator of checkpoint proteins that are upregulated in cancer. In some specific embodiments, the checkpoint inhibitor can be an inhibitor or activator of checkpoint proteins that include LAG-3, B7, TIM3 (HAVCR2), OX40 (CD134), GITR, CD137, CD40, VTCN1, IDO1, CD276, PVRIG, TIGIT, CD25 (IL2RA), IFNAR2, IFNAR1, CSF1R, VSIR (VISTA), or HLA. In some embodiments, the checkpoint inhibitor can be an inhibitor or activator selected from the group consisting of a PD-1 inhibitor, a PD-L1 inhibitor, a PD-L2 inhibitor, a CTLA-4 inhibitor, a LAG-3 inhibitor, a B7 inhibitor, a TIM3 (HAVCR2) inhibitor, an OX40 (CD134) inhibitor, a GITR agonist, a CD137 agonist, or a CD40 agonist, a VTCN1 inhibitor, an IDO1 inhibitor, a CD276 inhibitor, a PVRIG inhibitor, a TIGIT inhibitor, a CD25 (IL2RA) inhibitor, an IFNAR2 inhibitor, an IFNAR1 inhibitor, a CSF1R inhibitor, a VSIR (VISTA) inhibitor, or an HLA-targeted therapeutic agent. Such inhibitors, activators, or therapeutic agents are further described below.

[0220] In some embodiments, the checkpoint inhibitor is a CTLA-4 inhibitor. In one embodiment, the CTLA-4 inhibitor is an anti-CTLA-4 antibody. Examples of anti-CTLA-4 antibodies include, but are not limited to, those described in U.S. Patent Nos. 5,811,097, 5,811,097, 5,855,887, 6,051,227, 6,207,157, 6,682,736, 6,984,720, and 7,605,238 (all of which are incorporated herein in their entireties). In one embodiment, the anti-CTLA-4 antibody is tremelimumab (also known as ticilimumab or CP-675,206). In another embodiment, the anti-CTLA-4 antibody is ipilimumab (also known as MDX-010 or MDX-101). Ipilimumab is a fully human monoclonal IgG antibody that binds to CTLA-4. Ipilimumab is sold under the trade name Yervoy™.

[0221] In certain embodiments, the checkpoint inhibitor is a PD-1 / PD-L1 inhibitor. Examples of PD-1 / PD-L1 inhibitors include, but are not limited to, those described in U.S. Patent Nos. 7,488,802; 7,943,743; 8,008,449; 8,168,757; and 8,217,149, as well as PCT Patent Application Publication Nos. WO2003042402, WO2008156712, WO2010089411, WO2010036959, WO2011066342, WO2011159877, WO2011082400, and WO2011161699, all of which are incorporated herein in their entirety.

[0222] In some embodiments, the checkpoint inhibitor is a PD-1 inhibitor. In one embodiment, the PD-1 inhibitor is an anti-PD-1 antibody. In one embodiment, the anti-PD-1 antibody is BGB-A317, nivolumab (also known as ONO-4538, BMS-936558, or MDX1106), or pembrolizumab (also known as MK-3475, SCH900475, or lambrolizumab). In one embodiment, the anti-PD-1 antibody is nivolumab. Nivolumab is a human IgG4 anti-PD-1 monoclonal antibody and is sold under the trade name Opdivo™. In another embodiment, the anti-PD-1 antibody is pembrolizumab. Pembrolizumab is a humanized monoclonal IgG4 antibody and is sold under the trade name Keytruda™. In yet another embodiment, the anti-PD-1 antibody is the humanized antibody CT-011. CT-011 administered alone failed to elicit a response in the treatment of relapsed acute myeloid leukemia (AML). In yet another embodiment, the anti-PD-1 antibody is the fusion protein AMP-224. In another embodiment, the PD-1 antibody is BGB-A317. BGB-A317 is a monoclonal antibody specifically designed for binding to Fc gamma receptor I and possesses a unique binding signature for PD-1 with high affinity and excellent target specificity. In one embodiment, the PD-1 antibody is cemiplimab. In another embodiment, the PD-1 antibody is camrelizumab. In a further embodiment, the PD-1 antibody is sintilimab. In some embodiments, the PD-1 antibody is tislelizumab. In certain embodiments, the PD-1 antibody is TSR-042. In yet another embodiment, the PD-1 antibody is PDR001. In yet another embodiment, the PD-1 antibody is toripalimab.

[0223] In certain embodiments, the checkpoint inhibitor is a PD-L1 inhibitor. In one embodiment, the PD-L1 inhibitor is an anti-PD-L1 antibody. In one embodiment, the anti-PD-L1 antibody is MEDI4736 (durvalumab). In another embodiment, the anti-PD-L1 antibody is BMS-936559 (also known as MDX-1105-01). In yet another embodiment, the PD-L1 inhibitor is atezolizumab (also known as MPDL3280A and Tecentriq®). In a further embodiment, the PD-L1 inhibitor is avelumab.

[0224] In one embodiment, the checkpoint inhibitor is a PD-L2 inhibitor. In one embodiment, the PD-L2 inhibitor is an anti-PD-L2 antibody. In one embodiment, the anti-PD-L2 antibody is rHIgM12B7A.

[0225] In one embodiment, the checkpoint inhibitor is a lymphocyte activation gene-3 (LAG-3) inhibitor. In one embodiment, the LAG-3 inhibitor is IMP321, a soluble Ig fusion protein (Brignone et al., J. Immunol., 2007, 179, 4202-4211). In another embodiment, the LAG-3 inhibitor is BMS-986016.

[0226] In one embodiment, the checkpoint inhibitor is a B7 inhibitor. In one embodiment, the B7 inhibitor is a B7-H3 inhibitor or a B7-H4 inhibitor. In one embodiment, the B7-H3 inhibitor is MGA271, an anti-B7-H3 antibody (Loo et al., Clin. Cancer Res., 2012, 3834).

[0227] In one embodiment, the checkpoint inhibitor is a TIM3 (T-cell immunoglobulin domain and mucin domain 3) inhibitor (Fourcade et al., J. Exp. Med., 2010, 207, 2175-86; Sakuishi et al., J. Exp. Med., 2010, 207, 2187-94).

[0228] In one embodiment, the checkpoint inhibitor is an OX40 (CD134) agonist. In one embodiment, the checkpoint inhibitor is an anti-OX40 antibody. In one embodiment, the anti-OX40 antibody is anti-OX-40. In another embodiment, the anti-OX40 antibody is MEDI6469.

[0229] In one embodiment, the checkpoint inhibitor is a GITR agonist. In one embodiment, the checkpoint inhibitor is an anti-GITR antibody. In one embodiment, the anti-GITR antibody is TRX518.

[0230] In one embodiment, the checkpoint inhibitor is a CD137 agonist. In one embodiment, the checkpoint inhibitor is an anti-CD137 antibody. In one embodiment, the anti-CD137 antibody is urelumab. In another embodiment, the anti-CD137 antibody is PF-05082566.

[0231] In one embodiment, the checkpoint inhibitor is a CD40 agonist. In one embodiment, the checkpoint inhibitor is an anti-CD40 antibody. In one embodiment, the anti-CD40 antibody is CF-870,893.

[0232] In one embodiment, the checkpoint inhibitor is recombinant human interleukin-15 (rhIL-15).

[0233] In one embodiment, the checkpoint inhibitor is a VTCN inhibitor. In one embodiment, the VTCN inhibitor is FPA150.

[0234] In one embodiment, the checkpoint inhibitor is an IDO inhibitor. In one embodiment, the IDO inhibitor is INCB024360. In another embodiment, the IDO inhibitor is indoximod. In one embodiment, the IDO inhibitor is epacadostat. In another embodiment, the IDO inhibitor is BMS986205. In yet another embodiment, the IDO inhibitor is navoximod. In one embodiment, the IDO inhibitor is PF-06840003. In another embodiment, the IDO inhibitor is KHK2455. In yet another embodiment, the IDO inhibitor is RG70099. In one embodiment, the IDO inhibitor is IOM-E. In another embodiment, the IDO inhibitor is IOM-D.

[0235] In some embodiments, the checkpoint inhibitor is a TIGIT inhibitor. In certain embodiments, the TIGIT inhibitor is an anti-TIGIT antibody. In one embodiment, the TIGIT inhibitor is MTIG7192A. In another embodiment, the TIGIT inhibitor is BMS-986207. In yet another embodiment, the TIGIT inhibitor is OMP-313M32. In one embodiment, the TIGIT inhibitor is MK-7684. In another embodiment, the TIGIT inhibitor is AB154. In yet another embodiment, the TIGIT inhibitor is CGEN-15137. In one embodiment, the TIGIT inhibitor is SEA-TIGIT. In another embodiment, the TIGIT inhibitor is ASP8374. In yet another embodiment, the TIGIT inhibitor is AJUD008.

[0236] In some embodiments, the checkpoint inhibitor is a VSIR inhibitor. In certain embodiments, the VSIR inhibitor is an anti-VSIR antibody. In one embodiment, the VSIR inhibitor is MTIG7192A. In another embodiment, the VSIR inhibitor is CA-170. In yet another embodiment, the VSIR inhibitor is JNJ61610588. In one embodiment, the VSIR inhibitor is HMBD-002.

[0237] In some embodiments, the checkpoint inhibitor is a TIM3 inhibitor. In certain embodiments, the TIM3 inhibitor is an anti-TIM3 antibody. In one embodiment, the TIM3 inhibitor is AJUD009.

[0238] In some embodiments, the checkpoint inhibitor is a CD25 (IL2RA) inhibitor. In certain embodiments, the CD25 (IL2RA) inhibitor is an anti-CD25 (IL2RA) antibody. In one embodiment, the CD25 (IL2RA) inhibitor is daclizumab. In another embodiment, the CD25 (IL2RA) inhibitor is basiliximab.

[0239] In some embodiments, the checkpoint inhibitor is an IFNAR1 inhibitor. In certain embodiments, the IFNAR1 inhibitor is an anti-IFNAR1 antibody. In one embodiment, the IFNAR1 inhibitor is anifrolumab. In another embodiment, the IFNAR1 inhibitor is sifalimumab.

[0240] In some embodiments, the checkpoint inhibitor is a CSF1R inhibitor. In certain embodiments, the CSF1R inhibitor is an anti-CSF1R antibody. In one embodiment, the CSF1R inhibitor is pexidartinib. In another embodiment, the CSF1R inhibitor is emactuzumab. In yet another embodiment, the CSF1R inhibitor is caviralizumab. In one embodiment, the CSF1R inhibitor is ARRY-382. In another embodiment, the CSF1R inhibitor is BLZ945. In yet another embodiment, the CSF1R inhibitor is AJUD010. In one embodiment, the CSF1R inhibitor is AMG820. In another embodiment, the CSF1R inhibitor is IMC-CS4. In yet another embodiment, the CSF1R inhibitor is JNJ-40346527. In one embodiment, the CSF1R inhibitor is PLX5622. In another embodiment, the CSF1R inhibitor is FPA008.

[0241] In some embodiments, the checkpoint inhibitor is an HLA-targeted therapeutic agent. In certain embodiments, the HLA-targeted therapeutic agent is an anti-HLA antibody. In one embodiment, the HLA-targeted therapeutic agent is GSK01. In another embodiment, the HLA-targeted therapeutic agent is IMC-C103C. In yet another embodiment, the HLA-targeted therapeutic agent is IMC-F106C. In one embodiment, the HLA-targeted therapeutic agent is IMC-G107C. In another embodiment, the HLA-targeted therapeutic agent is ABBV-184.

[0242] In certain embodiments, the immune checkpoint inhibitors described herein include two or more of the checkpoint inhibitors described herein (including checkpoint inhibitors of the same or different classes).

[0243] In some embodiments, subjects that can be treated with the methods provided herein are mammals, hi some embodiments, subjects that can be treated with the methods provided herein are humans.

[0244] 5.2.1.4 Treatment Outcomes of the Methods Provided herein Despite the poor prognosis of cisplatin-ineligible human subjects who are frail as described above, suffer from multiple comorbidities other than their urothelial cancer / bladder cancer (e.g., muscle-invasive urothelial carcinoma (MIUC) and muscle-invasive bladder cancer (MIBC)), and are unable to tolerate additional treatments other than immunotherapy, the methods provided herein, including those described in this section (Section 5.2) and Sections 3 and 6, can provide beneficial treatment outcomes for these cisplatin-ineligible human subjects. In one embodiment, the human subject exhibits a complete response after treatment with the methods provided herein. In another embodiment, the human subject exhibits a partial response after treatment with the methods provided herein.

[0245] In some embodiments, response (complete or partial) is determined by evaluating the tumor or cancer site (lesion). The criteria for determining complete response (CR), partial response (PR), progressive disease (PD), and stable disease (SD) are listed in Table 15.

[0246] In some embodiments of the methods provided herein, the subject's pathological complete response rate (pCRR) is at least 30%. In some embodiments, the subject's pathological complete response rate (pCRR) is at least 35%. In some embodiments, the subject's pathological complete response rate (pCRR) is at least 40%. In some embodiments, the subject's pathological complete response rate (pCRR) is at least 45%. In some embodiments, the subject's pathological complete response rate (pCRR) is at least 50%. In some embodiments, the subject's pathological complete response rate (pCRR) is at least 55%. In some embodiments, the subject's pathological complete response rate (pCRR) is at least 60%. In some embodiments, the subject's pathological complete response rate (pCRR) is at least 65%. In some embodiments, the subject's pathological complete response rate (pCRR) is at least 70%. In some embodiments, the subject's pathological complete response rate (pCRR) is at least 75%.

[0247] In some embodiments of the methods provided herein, the pathological complete response rate (pCRR) is at least 30%. In some embodiments, the pathological complete response rate (pCRR) is at least 35%. In some embodiments, the pathological complete response rate (pCRR) is at least 40%. In some embodiments, the pathological complete response rate (pCRR) is at least 45%. In some embodiments, the pathological complete response rate (pCRR) is at least 50%. In some embodiments, the pathological complete response rate (pCRR) is at least 55%. In some embodiments, the pathological complete response rate (pCRR) is at least 60%. In some embodiments, the pathological complete response rate (pCRR) is at least 65%. In some embodiments, the pathological complete response rate (pCRR) is at least 70%. In some embodiments, the pathological complete response rate (pCRR) is at least 75%.

[0248] In some embodiments of the methods provided herein, the pathological complete response rate (pCRR) of a population of subjects treated with an ADC is at least 30%. In some embodiments, the pathological complete response rate (pCRR) of a population of subjects treated with an ADC is at least 35%. In some embodiments, the pathological complete response rate (pCRR) of a population of subjects treated with an ADC is at least 40%. In some embodiments, the pathological complete response rate (pCRR) of a population of subjects treated with an ADC is at least 45%. In some embodiments, the pathological complete response rate (pCRR) of a population of subjects treated with an ADC is at least 50%. In some embodiments, the pathological complete response rate (pCRR) of a population of subjects treated with an ADC is at least 55%. In some embodiments, the pathological complete response rate (pCRR) of a population of subjects treated with an ADC is at least 60%. In some embodiments, the pathological complete response rate (pCRR) of a population of subjects treated with an ADC is at least 65%. In some embodiments, the pathological complete response rate (pCRR) in a population of subjects treated with the ADC is at least 70%. In some embodiments, the pathological complete response rate (pCRR) in a population of subjects treated with the ADC is at least 75%.

[0249] In some embodiments of the methods provided herein, the human subject has at least or about 2 months of disease-free survival after treatment. In another embodiment, the human subject has at least or about 3 months of disease-free survival after treatment. In another embodiment, the human subject has at least or about 4 months of disease-free survival after treatment. In another embodiment, the human subject has at least or about 5 months of disease-free survival after treatment. In another embodiment, the human subject has at least or about 6 months of disease-free survival after treatment. In a further embodiment, the human subject has at least or about 7 months of disease-free survival after treatment. In yet another embodiment, the human subject has at least or about 8 months of disease-free survival after treatment. In one embodiment, the human subject has at least or about 9 months of disease-free survival after treatment. In another embodiment, the human subject has at least or about 10 months of disease-free survival after treatment. In a further embodiment, the human subject has at least or about 11 months of disease-free survival after treatment. In yet another embodiment, the human subject has at least or about 12 months of disease-free survival after treatment. In one embodiment, the human subject has a disease-free survival of at least or about 13 months after treatment. In another embodiment, the human subject has a disease-free survival of at least or about 14 months after treatment. In yet another embodiment, the human subject has a disease-free survival of at least or about 15 months after treatment. In some embodiments, the subject's overall survival is extended by at least 2, at least 4, at least 6, at least 8, at least 10, or at least 12 months.

[0250] In one embodiment, the human subject has a disease-free survival period ranging from 2 to 15 months after treatment. In another embodiment, the human subject has a disease-free survival period ranging from 2 to 14 months after treatment. In a further embodiment, the human subject has a disease-free survival period ranging from 2 to 13 months after treatment. In one embodiment, the human subject has a disease-free survival period ranging from 2 to 12 months after treatment. In another embodiment, the human subject has a disease-free survival period ranging from 3 to 12 months after treatment. In a further embodiment, the human subject has a disease-free survival period ranging from 3 to 11 months after treatment. In yet another embodiment, the human subject has a disease-free survival period ranging from 4 to 11 months after treatment. In one embodiment, the human subject has a disease-free survival period ranging from 4 to 10 months after treatment. In another embodiment, the human subject has a disease-free survival period ranging from 5 to 10 months after treatment. In a further embodiment, the human subject has a disease-free survival period ranging from 5 to 9 months after treatment. In yet another embodiment, the human subject has a disease-free survival period ranging from 5 to 8 months after treatment. In one embodiment, the human subject has a disease-free survival period ranging from 5 to 7 months after treatment. In one embodiment, the human subject has a disease-free survival period ranging from 5 to 6 months after treatment. In one embodiment, the human subject has a disease-free survival period ranging from 6 to 7 months after treatment. In another embodiment, the human subject has a disease-free survival period ranging from 6 to 8 months after treatment.

[0251] Additionally, in some embodiments, disease-free survival is assessed for a population of human subjects treated by the methods provided herein by assessing the median or mean disease-free survival in the treated population. In one embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean disease-free survival in the treated population is at least or about 2 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean disease-free survival in the treated population is at least or about 3 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean disease-free survival in the treated population is at least or about 4 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean disease-free survival in the treated population is at least or about 5 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean disease-free survival in the treated population is at least or about 6 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean disease-free survival in the treated population is at least or about 7 months. In yet another embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean disease-free survival in the treated population is at least or about 8 months. In one embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean disease-free survival in the treated population is at least or about 9 months. In another embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean disease-free survival in the treated population is at least or about 10 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean disease-free survival in the treated population is at least or about 11 months.In yet another embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean disease-free survival in the treated population is at least or about 12 months. In one embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean disease-free survival in the treated population is at least or about 13 months. In another embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean disease-free survival in the treated population is at least or about 14 months. In yet another embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean disease-free survival in the treated population is at least or about 15 months.

[0252] In one embodiment, a population of human subjects is treated by the methods provided herein, and the disease-free survival in the treated population ranges from 2 to 15 months. In another embodiment, a population of human subjects is treated by the methods provided herein, and the disease-free survival in the treated population ranges from 2 to 14 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the disease-free survival in the treated population ranges from 2 to 13 months. In yet another embodiment, a population of human subjects is treated by the methods provided herein, and the disease-free survival in the treated population ranges from 2 to 12.32 months. In one embodiment, a population of human subjects is treated by the methods provided herein, and the disease-free survival in the treated population ranges from 2 to 12 months. In another embodiment, a population of human subjects is treated by the methods provided herein, and the disease-free survival in the treated population ranges from 3 to 12 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the disease-free survival in the treated population ranges from 3 to 11 months. In yet another embodiment, a population of human subjects is treated by the methods provided herein, and the disease-free survival in the treated population ranges from 4 to 11 months. In one embodiment, a population of human subjects is treated by the methods provided herein, and the disease-free survival in the treated population ranges from 4 to 10 months. In another embodiment, a population of human subjects is treated by the methods provided herein, and the disease-free survival in the treated population ranges from 5 to 10 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the disease-free survival in the treated population ranges from 5 to 9 months. In one embodiment, a population of human subjects is treated by the methods provided herein, and the disease-free survival in the treated population ranges from 5 to 8 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the disease-free survival in the treated population ranges from 5 to 7 months.In a further embodiment, a population of human subjects is treated with the methods provided herein and disease-free survival in the treated population ranges from 5 to 6 months. In one embodiment, a population of human subjects is treated with the methods provided herein and disease-free survival in the treated population ranges from 6 to 7 months. In yet another embodiment, a population of human subjects is treated with the methods provided herein and disease-free survival in the treated population ranges from 6 to 8 months.

[0253] In some embodiments of the methods provided herein, the human subject has a recurrence-free survival of at least or about 2 months after treatment. In another embodiment, the human subject has a recurrence-free survival of at least or about 2.6 months after treatment. In another embodiment, the human subject has a recurrence-free survival of at least or about 3 months after treatment. In another embodiment, the human subject has a recurrence-free survival of at least or about 4 months after treatment. In another embodiment, the human subject has a recurrence-free survival of at least or about 5 months after treatment. In another embodiment, the human subject has a recurrence-free survival of at least or about 6 months after treatment. In another embodiment, the human subject has a recurrence-free survival of at least or about 6.54 months after treatment. In a further embodiment, the human subject has a recurrence-free survival of at least or about 7 months after treatment. In yet another embodiment, the human subject has a recurrence-free survival of at least or about 8 months after treatment. In one embodiment, the human subject has a recurrence-free survival of at least or about 9 months after treatment. In another embodiment, the human subject has a recurrence-free survival of at least or about 10 months after treatment. In a further embodiment, the human subject has a recurrence-free survival of at least or about 11 months after treatment. In yet another embodiment, the human subject has a recurrence-free survival of at least or about 12 months after treatment. In yet another embodiment, the human subject has a recurrence-free survival of at least or about 12.32 months after treatment. In one embodiment, the human subject has a recurrence-free survival of at least or about 13 months after treatment. In another embodiment, the human subject has a recurrence-free survival of at least or about 14 months after treatment. In yet another embodiment, the human subject has a recurrence-free survival of at least or about 15 months after treatment.

[0254] In one embodiment, the human subject has a recurrence-free survival period ranging from 2 to 15 months after treatment. In another embodiment, the human subject has a recurrence-free survival period ranging from 2 to 14 months after treatment. In a further embodiment, the human subject has a recurrence-free survival period ranging from 2 to 13 months after treatment. In yet another embodiment, the human subject has a recurrence-free survival period ranging from 2 to 12.32 months after treatment. In one embodiment, the human subject has a recurrence-free survival period ranging from 2 to 12 months after treatment. In another embodiment, the human subject has a recurrence-free survival period ranging from 3 to 12 months after treatment. In a further embodiment, the human subject has a recurrence-free survival period ranging from 3 to 11 months after treatment. In yet another embodiment, the human subject has a recurrence-free survival period ranging from 4 to 11 months after treatment. In one embodiment, the human subject has a recurrence-free survival period ranging from 4 to 10 months after treatment. In another embodiment, the human subject has a recurrence-free survival period ranging from 5 to 10 months after treatment. In a further embodiment, the human subject has a recurrence-free survival period ranging from 5 to 9 months after treatment. In yet another embodiment, the human subject has a recurrence-free survival period ranging from 5 to 8 months after treatment. In one embodiment, the human subject has a recurrence-free survival period ranging from 5 to 7 months after treatment. In one embodiment, the human subject has a recurrence-free survival period ranging from 5 to 6 months after treatment. In one embodiment, the human subject has a recurrence-free survival period ranging from 6 to 7 months after treatment. In another embodiment, the human subject has a recurrence-free survival period ranging from 6 to 8 months after treatment.

[0255] Additionally, in some embodiments, recurrence-free survival is assessed for a population of human subjects treated by the methods provided herein by assessing the median or mean recurrence-free survival in the treated population. In one embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean recurrence-free survival in the treated population is at least or about 2 months. In another embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean recurrence-free survival in the treated population is at least or about 2.6 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean recurrence-free survival in the treated population is at least or about 3 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean recurrence-free survival in the treated population is at least or about 4 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean recurrence-free survival in the treated population is at least or about 5 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean recurrence-free survival in the treated population is at least or about 6 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean recurrence-free survival in the treated population is at least or about 7 months. In yet another embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean recurrence-free survival in the treated population is at least or about 8 months. In one embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean recurrence-free survival in the treated population is at least or about 9 months. In another embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean recurrence-free survival in the treated population is at least or about 10 months.In a further embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean recurrence-free survival in the treated population is at least or about 11 months. In yet another embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean recurrence-free survival in the treated population is at least or about 12 months. In yet another embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean recurrence-free survival in the treated population is at least or about 12.32 months. In one embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean recurrence-free survival in the treated population is at least or about 13 months. In another embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean recurrence-free survival in the treated population is at least or about 14 months. In yet another embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean recurrence-free survival in the treated population is at least or about 15 months.

[0256] In one embodiment, a population of human subjects is treated by the methods provided herein, and the recurrence-free survival in the treated population ranges from 2 to 15 months. In another embodiment, a population of human subjects is treated by the methods provided herein, and the recurrence-free survival in the treated population ranges from 2 to 14 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the recurrence-free survival in the treated population ranges from 2 to 13 months. In yet another embodiment, a population of human subjects is treated by the methods provided herein, and the recurrence-free survival in the treated population ranges from 2 to 12.32 months. In one embodiment, a population of human subjects is treated by the methods provided herein, and the recurrence-free survival in the treated population ranges from 2 to 12 months. In another embodiment, a population of human subjects is treated by the methods provided herein, and the recurrence-free survival in the treated population ranges from 3 to 12 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the recurrence-free survival in the treated population ranges from 3 to 11 months. In yet another embodiment, a population of human subjects is treated by the methods provided herein, and the recurrence-free survival in the treated population ranges from 4 to 11 months. In one embodiment, a population of human subjects is treated by the methods provided herein, and the recurrence-free survival in the treated population ranges from 4 to 10 months. In another embodiment, a population of human subjects is treated by the methods provided herein, and the recurrence-free survival in the treated population ranges from 5 to 10 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the recurrence-free survival in the treated population ranges from 5 to 9 months. In one embodiment, a population of human subjects is treated by the methods provided herein, and the recurrence-free survival in the treated population ranges from 5 to 8 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the recurrence-free survival in the treated population ranges from 5 to 7 months.In a further embodiment, a population of human subjects is treated with the methods provided herein, and recurrence-free survival in the treated population ranges from 5 to 6 months. In one embodiment, a population of human subjects is treated with the methods provided herein, and recurrence-free survival in the treated population ranges from 6 to 7 months. In yet another embodiment, a population of human subjects is treated with the methods provided herein, and recurrence-free survival in the treated population ranges from 6 to 8 months.

[0257] In some embodiments of the methods provided herein, the human subject has an overall survival of at least or about 5 months after treatment. In another embodiment, the human subject has an overall survival of at least or about 6 months after treatment. In a further embodiment, the human subject has an overall survival of at least or about 7 months after treatment. In yet another embodiment, the human subject has an overall survival of at least or about 8 months after treatment. In one embodiment, the human subject has an overall survival of at least or about 9 months after treatment. In another embodiment, the human subject has an overall survival of at least or about 10 months after treatment. In a further embodiment, the human subject has an overall survival of at least or about 11 months after treatment. In yet another embodiment, the human subject has an overall survival of at least or about 12 months after treatment. In one embodiment, the human subject has an overall survival of at least or about 13 months after treatment. In another embodiment, the human subject has an overall survival of at least or about 14 months after treatment. In yet another embodiment, the human subject has an overall survival of at least or about 15 months after treatment. In one embodiment, the human subject has an overall survival of at least or about 16 months after treatment. In another embodiment, the human subject has an overall survival of at least or about 17 months after treatment. In a further embodiment, the human subject has an overall survival of at least or about 18 months after treatment. In yet another embodiment, the human subject has an overall survival of at least or about 19 months after treatment. In one embodiment, the human subject has an overall survival of at least or about 20 months after treatment. In another embodiment, the human subject has an overall survival of at least or about 21 months after treatment. In a further embodiment, the human subject has an overall survival of at least or about 22 months after treatment. In a further embodiment, the human subject has an overall survival of at least or about 23 months after treatment. In yet another embodiment, the human subject has an overall survival of at least or about 24 months after treatment. In one embodiment, the human subject has an overall survival of at least or about 25 months after treatment. In another embodiment, the human subject has an overall survival of at least or about 26 months after treatment.In a further embodiment, the human subject has an overall survival of at least or about 27 months after treatment. In one embodiment, the human subject has an overall survival of at least or about 28 months after treatment. In another embodiment, the human subject has an overall survival of at least or about 29 months after treatment. In a further embodiment, the human subject has an overall survival of at least or about 30 months after treatment.

[0258] In one embodiment, the human subject has an overall survival time in the range of 10 to 19 months after treatment. In another embodiment, the human subject has an overall survival time in the range of 10 to 18 months after treatment. In a further embodiment, the human subject has an overall survival time in the range of 10 to 17 months after treatment. In yet another embodiment, the human subject has an overall survival time in the range of 10 to 16 months after treatment. In one embodiment, the human subject has an overall survival time in the range of 10 to 15 months after treatment. In another embodiment, the human subject has an overall survival time in the range of 10 to 14 months after treatment. In a further embodiment, the human subject has an overall survival time in the range of 10 to 13 months after treatment. In yet another embodiment, the human subject has an overall survival time in the range of 10 to 12 months after treatment. In one embodiment, the human subject has an overall survival time in the range of 10 to 11 months after treatment. In another embodiment, the human subject has an overall survival time in the range of 11 to 19 months after treatment. In a further embodiment, the human subject has an overall survival time in the range of 12 to 19 months after treatment. In yet another embodiment, the human subject has an overall survival time in the range of 13 to 19 months after treatment. In one embodiment, the human subject has an overall survival time in the range of 14 to 18 months after treatment. In one embodiment, the human subject has an overall survival time in the range of 14 to 19 months after treatment. In one embodiment, the human subject has an overall survival time in the range of 15 to 18 months after treatment. In another embodiment, the human subject has an overall survival time in the range of 15 to 19 months after treatment. In a further embodiment, the human subject has an overall survival time in the range of 16 to 19 months after treatment. In yet another embodiment, the human subject has an overall survival time in the range of 17 to 19 months after treatment. In one embodiment, the human subject has an overall survival time in the range of 18 to 19 months after treatment. In another embodiment, the human subject has an overall survival time in the range of 11 to 18 months after treatment. In a further embodiment, the human subject has an overall survival time in the range of 12 to 17 months after treatment. In a further embodiment, the human subject has an overall survival time in the range of 13 to 16 months after treatment. In yet another embodiment, the human subject has an overall survival time in the range of 14 to 15 months after treatment, hi one embodiment, the human subject has an overall survival time in the range of 10 to 20 months after treatment.In another embodiment, the human subject has an overall survival time in the range of 11 to 20 months after treatment. In one embodiment, the human subject has an overall survival time in the range of 11 to 24 months after treatment. In one embodiment, the human subject has an overall survival time in the range of 11 to 25 months after treatment. In one embodiment, the human subject has an overall survival time in the range of 12 to 24 months after treatment. In one embodiment, the human subject has an overall survival time in the range of 12 to 25 months after treatment. In a further embodiment, the human subject has an overall survival time in the range of 12 to 20 months after treatment. In one embodiment, the human subject has an overall survival time in the range of 13 to 20 months after treatment. In another embodiment, the human subject has an overall survival time in the range of 14 to 20 months after treatment. In a further embodiment, the human subject has an overall survival time in the range of 15 to 20 months after treatment. In yet another embodiment, the human subject has an overall survival time in the range of 16 to 20 months after treatment. In one embodiment, the human subject has an overall survival time in the range of 17 to 20 months after treatment. In another embodiment, the human subject has an overall survival time in the range of 18 to 20 months after treatment. In a further embodiment, the human subject has an overall survival time in the range of 19 to 20 months after treatment. In one embodiment, the human subject has an overall survival time in the range of 9 to 20 months after treatment. In another embodiment, the human subject has an overall survival time in the range of 9 to 19 months after treatment. In a further embodiment, the human subject has an overall survival time in the range of 9 to 18 months after treatment. In one embodiment, the human subject has an overall survival time in the range of 9 to 17 months after treatment. In another embodiment, the human subject has an overall survival time in the range of 9 to 16 months after treatment. In a further embodiment, the human subject has an overall survival time in the range of 9 to 15 months after treatment. In one embodiment, the human subject has an overall survival time in the range of 9 to 14 months after treatment. In another embodiment, the human subject has an overall survival time in the range of 9 to 13 months after treatment. In a further embodiment, the human subject has an overall survival time in the range of 9 to 12 months after treatment. In one embodiment, the human subject has an overall survival time in the range of 9 to 11 months after treatment. In another embodiment, the human subject has an overall survival after treatment in the range of 9 to 10 months.

[0259] Additionally, in some embodiments, overall survival is assessed for a population of human subjects treated by the methods provided herein by assessing the median or mean overall survival in the treated population. In one embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean overall survival in the treated population is at least or about 5 months. In another embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean overall survival in the treated population is at least or about 6 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean overall survival in the treated population is at least or about 7 months. In yet another embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean overall survival in the treated population is at least or about 8 months. In one embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean overall survival in the treated population is at least or about 9 months. In another embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean overall survival in the treated population is at least or about 10 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean overall survival in the treated population is at least or about 11 months. In yet another embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean overall survival in the treated population is at least or about 12 months. In one embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean overall survival in the treated population is at least or about 13 months. In another embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean overall survival in the treated population is at least or about 14 months.In yet another embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean overall survival in the treated population is at least or about 15 months. In one embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean overall survival in the treated population is at least or about 16 months. In another embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean overall survival in the treated population is at least or about 17 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean overall survival in the treated population is at least or about 18 months. In yet another embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean overall survival in the treated population is at least or about 19 months. In one embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean overall survival in the treated population is at least or about 20 months. In another embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean overall survival in the treated population is at least or about 21 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean overall survival in the treated population is at least or about 22 months. In yet another embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean overall survival in the treated population is at least or about 23 months. In one embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean overall survival in the treated population is at least or about 24 months. In another embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean overall survival in the treated population is at least or about 25 months.In a further embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean overall survival in the treated population is at least or about 26 months. In yet another embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean overall survival in the treated population is at least or about 27 months. In one embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean overall survival in the treated population is at least or about 28 months. In another embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean overall survival in the treated population is at least or about 29 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the median or mean overall survival in the treated population is at least or about 30 months.

[0260] In one embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 10 to 19 months. In another embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 10 to 18 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 10 to 17 months. In yet another embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 10 to 16 months. In one embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 10 to 15 months. In another embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 10 to 14 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 10 to 13 months. In yet another embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population is in the range of 10 to 12 months. In one embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population is in the range of 10 to 11 months. In another embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population is in the range of 10 to 19 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population is in the range of 11 to 19 months. In one embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population is in the range of 11 to 24 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population is in the range of 11 to 25 months.In a further embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 12 to 24 months. In one embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 12 to 25 months. In yet another embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 12 to 19 months. In one embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 13 to 19 months. In another embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 14 to 19 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 15 to 19 months. In yet another embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 16 to 19 months. In one embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 17 to 19 months. In another embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 18 to 19 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 11 to 18 months. In yet another embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 12 to 17 months. In one embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 13 to 16 months. In another embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 14 to 15 months.In a further embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population is in the range of 10 to 20 months. In yet another embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population is in the range of 11 to 20 months. In one embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population is in the range of 12 to 20 months. In another embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population is in the range of 13 to 20 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population is in the range of 14 to 20 months. In yet another embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population is in the range of 15 to 20 months. In one embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population is in the range of 16 to 20 months. In another embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 17 to 20 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 18 to 20 months. In yet another embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 19 to 20 months. In one embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 9 to 20 months. In another embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 9 to 19 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 9 to 18 months.In yet another embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 9 to 17 months. In one embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 9 to 16 months. In another embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 9 to 15 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 9 to 14 months. In yet another embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 9 to 13 months. In one embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 9 to 12 months. In another embodiment, a population of human subjects is treated by the methods provided herein, and the overall survival in the treated population ranges from 9 to 11 months. In a further embodiment, a population of human subjects is treated by the methods provided herein, and overall survival in the treated population ranges from 9 to 10 months.

[0261] In some embodiments of the methods provided herein, the subject's pathological downstaging rate (pDSR) is at least 50%. In some embodiments, the subject's pathological downstaging rate (pDSR) is at least 55%. In some embodiments, the subject's pathological downstaging rate (pDSR) is at least 60%. In some embodiments, the subject's pathological downstaging rate (pDSR) is at least 65%. In some embodiments, the subject's pathological downstaging rate (pDSR) is at least 70%. In some embodiments, the subject's pathological downstaging rate (pDSR) is at least 75%. In some embodiments, the subject's pathological downstaging rate (pDSR) is at least 80%. In some embodiments, the subject's pathological downstaging rate (pDSR) is at least 85%. In some embodiments, the subject's pathological downstaging rate (pDSR) is at least 90%.

[0262] In some embodiments of the methods provided herein, the pathological downstaging rate (pDSR) is at least 50%. In some embodiments, the pathological downstaging rate (pDSR) is at least 55%. In some embodiments, the pathological downstaging rate (pDSR) is at least 60%. In some embodiments, the pathological downstaging rate (pDSR) is at least 65%. In some embodiments, the pathological downstaging rate (pDSR) is at least 70%. In some embodiments, the pathological downstaging rate (pDSR) is at least 75%. In some embodiments, the pathological downstaging rate (pDSR) is at least 80%. In some embodiments, the pathological downstaging rate (pDSR) is at least 85%. In some embodiments, the pathological downstaging rate (pDSR) is at least 90%.

[0263] In some embodiments of the methods provided herein, the pathological downstaging rate (pDSR) of a population of subjects treated with an ADC is at least 50%. In some embodiments, the pathological downstaging rate (pDSR) of a population of subjects treated with an ADC is at least 55%. In some embodiments, the pathological downstaging rate (pDSR) of a population of subjects treated with an ADC is at least 60%. In some embodiments, the pathological downstaging rate (pDSR) of a population of subjects treated with an ADC is at least 65%. In some embodiments, the pathological downstaging rate (pDSR) of a population of subjects treated with an ADC is at least 70%. In some embodiments, the pathological downstaging rate (pDSR) of a population of subjects treated with an ADC is at least 75%.

[0264] In some embodiments, human subject and patient are used interchangeably. Thus, one of skill in the art will understand that a human subject can be interchangeable with a patient in any of the methods provided herein.

[0265] 5.2.2 Methods for Treating Cancer in Patient Populations Based on Additional Selection Criteria Provided herein are methods for treating various cancers in a subject, wherein the cancer has any of the suitable markers and / or characteristics as provided in Section 6. Also provided herein are methods for treating various cancers in a subject, wherein the subject has any of the suitable characteristics as provided in Section 6.

[0266] In one aspect, provided herein is a method of preventing or treating cancer in a subject, comprising administering to the subject an effective amount of an antibody drug conjugate, wherein the antibody drug conjugate comprises an antibody or antigen-binding fragment thereof that binds to 191P4D12 conjugated to one or more units of monomethyl auristatin E (MMAE), and wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising a complementarity-determining region (CDR) comprising the amino acid sequence of the heavy chain variable region CDR set forth in SEQ ID NO: 22 and a light chain variable region comprising a CDR comprising the amino acid sequence of the light chain variable region CDR set forth in SEQ ID NO: 23, and wherein the subject has any of the suitable characteristics as provided in Section 6.

[0267] In some aspects, provided herein are methods of preventing or treating cancer in a subject, comprising administering to the subject an effective amount of an antibody drug conjugate, wherein the antibody drug conjugate comprises an antibody or antigen-binding fragment thereof that binds to 191P4D12 conjugated to one or more units of monomethyl auristatin E (MMAE), wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising a complementarity-determining region (CDR) comprising the amino acid sequence of the heavy chain variable region CDR set forth in SEQ ID NO: 22 and a light chain variable region comprising a CDR comprising the amino acid sequence of the light chain variable region CDR set forth in SEQ ID NO: 23, and wherein the cancer has any of the suitable markers and / or characteristics as provided in Section 6.

[0268] In another aspect, provided herein is a method of preventing or treating cancer in a subject, comprising administering to the subject an effective amount of an antibody drug conjugate, wherein the antibody drug conjugate comprises an antibody or antigen-binding fragment thereof that binds to 191P4D12 conjugated to one or more units of monomethyl auristatin E (MMAE), and wherein the subject has any of the suitable characteristics as provided in Section 6. In a further aspect, provided herein is a method of preventing or treating cancer in a subject, comprising administering to the subject an effective amount of an antibody drug conjugate, wherein the antibody drug conjugate comprises an antibody or antigen-binding fragment thereof that binds to 191P4D12 conjugated to one or more units of monomethyl auristatin E (MMAE), and wherein the cancer has any of the suitable markers and / or characteristics as provided in Section 6.

[0269] In some embodiments of the methods provided herein, including Section 5.2, including Sections 5.2.1 and 5.2.2, and Sections 3 and 6, the subject is a human subject.

[0270] All of the methods provided herein, and specifically those described in Sections 5.2.1 and 5.2.2: Therapeutic agents, including ADCs, that may be used are described in Sections 3, 5.2, 5.3, 5.4, 5.5, and 6; patient selection for treatment is described herein and exemplified in Section 5.2, including Sections 5.2.1 and 5.2.2, and Sections 3 and 6; dosing regimens and pharmaceutical compositions for administering therapeutic agents are described in Sections 5.4, 5.7, and in Section 6 below; and biomarkers that can be used to identify therapeutic agents, select patients, determine outcome of these methods, and / or serve as criteria in any manner for these methods are described herein and in Section 5.2. Biomarkers are exemplified in Section 5.2, including Sections 5.1 and 5.2.2, and Sections 3 and 6; biomarkers can be determined as described in Section 5.8 or as known in the art; treatment outcomes for the methods provided herein are described in this section (Section 5.2, including Section 5.2.1.4) and Sections 3 and 6; additional treatment outcomes for the methods provided herein can be improvement in the biomarkers described herein, e.g., the biomarkers described and exemplified in Section 5.2, including Sections 5.2.1 and 5.2.2, and Sections 3 and 6; combination therapies including ADCs and other therapeutic agents are described in this section (Section 5.2) and Section 5.5. Accordingly, one of skill in the art will understand that the methods provided herein include all permutations and combinations of patients, therapeutic agents, dosing regimens, biomarkers, and treatment outcomes as described above and below.

[0271] 5.3 Antibody Drug Conjugates for the Method In various embodiments of the methods provided herein, including those provided in Section 5.2, the ADC used in the method comprises or is an anti-191P4D12 ADC described herein and / or in U.S. Patent No. 8,637,642, which is incorporated herein by reference in its entirety. In some embodiments, the anti-191P4D12 antibody drug conjugates provided for the methods herein comprise an antibody or antigen-binding fragment thereof that binds to 191P4D12 as provided herein, including Sections 3, 5.3.1, and 6, and are conjugated to one or more units of a cytotoxic agent (Drug unit, or D) as provided herein, including Sections 3 and 6 and this section (Section 5.3), with further disclosure in Sections 5.3.2 and 5.3.4. In certain embodiments, the cytotoxic agent (Drug unit, or D) can be covalently attached directly or via a linker unit (LU) as provided herein, including Sections 3 and 6 and this section (Section 5.3), with further disclosure in Section 5.3.3.

[0272] In some embodiments, the antibody drug conjugate compound has the following formula: L-(LU-D) p (I) or a pharmaceutically acceptable salt or solvate thereof, wherein L is an antibody unit, e.g., as provided in Sections 3, 5.3.1, and 6, e.g., an anti-Nectin-4 antibody or an antigen-binding fragment thereof; (LU-D) is the Linker Unit-Drug Unit moiety, where: LU- is a linker unit, e.g., as provided in Sections 3 and 6 and this section (Section 5.3), with further disclosure in Section 5.3.3; D is a drug entity having cytostatic or cytotoxic activity against target cells, e.g., as provided in Sections 3 and 6 and this section (Section 5.3), with further disclosure in Sections 5.3.2 and 5.3.4; p is an integer between 1 and 20, with further examples provided in Sections 3 and 6 and this section (Section 5.3).

[0273] In some embodiments, p is in the range of 1 to 20, 1 to 19, 1 to 18, 1 to 17, 1 to 16, 1 to 15, 1 to 14, 1 to 13, 1 to 12, 1 to 11, 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, or 1 to 2. In some embodiments, p is in the range of 2 to 20, 2 to 19, 2 to 18, 2 to 17, 2 to 16, 2 to 15, 2 to 14, 2 to 13, 2 to 12, 2 to 11, 2 to 10, 2 to 9, 2 to 8, 2 to 7, 2 to 6, 2 to 5, 2 to 4, or 2 to 3. In some embodiments, p is in the range of 3 to 20, 3 to 19, 3 to 18, 3 to 17, 3 to 16, 3 to 15, 3 to 14, 3 to 13, 3 to 12, 3 to 11, 3 to 10, 3 to 9, 3 to 8, 3 to 7, 3 to 6, 3 to 5, or 3 to 4. In some embodiments, p is about 1. In some embodiments, p is about 2. In some embodiments, p is about 3. In some embodiments, p is about 4. In some embodiments, p is about 3.8. In some embodiments, p is about 5. In some embodiments, p is about 6. In some embodiments, p is about 7. In some embodiments, p is about 8. In some embodiments, p is about 9. In some embodiments, p is about 10. In some embodiments, p is about 11. In some embodiments, p is about 12. In some embodiments, p is about 13. In some embodiments, p is about 14. In some embodiments, p is about 15. In some embodiments, p is about 16. In some embodiments, p is about 17. In some embodiments, p is about 18. In some embodiments, p is about 19. In some embodiments, p is about 20.

[0274] In some embodiments, the antibody drug conjugate compound has the following formula: L-(A a -W w -Y y -D) p(II) or a pharmaceutically acceptable salt or solvate thereof, wherein L is an antibody unit, e.g., as provided in Sections 3, 5.3.1, and 6, e.g., an anti-Nectin-4 antibody or an antigen-binding fragment thereof; -A a -W w -Y y - is a linker unit (LU), where -A- is a stretcher unit, a is 0 or 1, each -W- is independently an amino acid unit; w is an integer ranging from 0 to 12, -Y- is a self-immolative spacer unit; y is 0, 1, or 2; Each is provided, for example, in Sections 3 and 6 and in this section (Section 5.3), with further disclosure in Section 5.3.3. D is a drug entity having cytostatic or cytotoxic activity against target cells, e.g., as provided in Sections 3 and 6 and this section (Section 5.3), with further disclosure in Sections 5.3.2 and 5.3.4; p is an integer between 1 and 20, with further examples provided in Sections 3 and 6 and this section (Section 5.3).

[0275] In some embodiments, a is 0 or 1, w is 0 or 1, and y is 0, 1, or 2. In some embodiments, a is 0 or 1, w is 0 or 1, and y is 0 or 1. In some embodiments, p is in the range of 1 to 20, 1 to 19, 1 to 18, 1 to 17, 1 to 16, 1 to 15, 1 to 14, 1 to 13, 1 to 12, 1 to 11, 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, or 1 to 2. In some embodiments, p is in the range of 2 to 20, 2 to 19, 2 to 18, 2 to 17, 2 to 16, 2 to 15, 2 to 14, 2 to 13, 2 to 12, 2 to 11, 2 to 10, 2 to 9, 2 to 8, 2 to 7, 2 to 6, 2 to 5, 2 to 4, or 2 to 3. In some embodiments, p is in the range of 3 to 20, 3 to 19, 3 to 18, 3 to 17, 3 to 16, 3 to 15, 3 to 14, 3 to 13, 3 to 12, 3 to 11, 3 to 10, 3 to 9, 3 to 8, 3 to 7, 3 to 6, 3 to 5, or 3 to 4. In some embodiments, p is about 1. In some embodiments, p is about 2. In some embodiments, p is about 3. In some embodiments, p is about 4. In some embodiments, p is about 3.8. In some embodiments, p is about 5. In some embodiments, p is about 6. In some embodiments, p is about 7. In some embodiments, p is about 8. In some embodiments, p is about 9. In some embodiments, p is about 10. In some embodiments, p is about 11. In some embodiments, p is about 12. In some embodiments, p is about 13. In some embodiments, p is about 14. In some embodiments, p is about 15. In some embodiments, p is about 16. In some embodiments, p is about 17. In some embodiments, p is about 18. In some embodiments, p is about 19. In some embodiments, p is about 20. In some embodiments, when w is not zero, y is 1 or 2. In some embodiments, when w is 1-12, y is 1 or 2. In some embodiments, w is 2-12 and y is 1 or 2. In some embodiments, a is 1 and w and y are 0.

[0276] In some specific embodiments of the methods provided herein, including those provided in Section 5.2, the cytotoxic agent as part of any of the ADCs provided herein for the methods comprises, consists of, or is MMAE.

[0277] For compositions comprising multiple antibodies or antigen-binding fragments thereof, drug loading is represented by p, the average number of drug molecules per antibody unit. Drug loading can range from 1 to 20 drugs (D) per antibody. The average number of drugs per antibody in preparation for conjugation reactions can be characterized by conventional means, such as mass spectrometry, ELISA assay, and HPLC. The quantitative distribution of antibody-drug conjugates with respect to p can also be determined. In some cases, separation, purification, and characterization of homogeneous antibody-drug conjugates with a constant p value from antibody-drug conjugates with other drug loads can be achieved by means such as reverse-phase HPLC or electrophoresis. In certain exemplary embodiments, p is 2 to 8.

[0278] Additional embodiments of ADCs for the methods provided herein are described in U.S. Pat. No. 8,637,642 and International Application No. PCT / US2019 / 056214 (Publication No. WO2020 / 117373), both of which are incorporated by reference herein in their entireties.

[0279] In some embodiments of the methods provided herein, including Sections 3, 5.2, and 6, and this section (Section 5.3), the ADC is enfortumab vedotin. In certain embodiments of the methods provided herein, including Sections 3, 5.2, and 6, and this section (Section 5.3), the ADC is a biosimilar of enfortumab vedotin.

[0280] In some embodiments of the methods provided herein, the ADC is administered as a monotherapy.

[0281] 5.3.1 Anti-191P4D12 Antibodies or Antigen-Binding Fragments In one embodiment, the antibody or antigen-binding fragment that binds to a Nectin-4-related protein is an antibody or antigen-binding fragment that specifically binds to a Nectin-4 protein comprising the amino acid sequence of SEQ ID NO: 2 (see Figure 1A). The corresponding cDNA encoding the 191P4D12 protein has the sequence of SEQ ID NO: 1 (see Figure 1A).

[0282] Antibodies that specifically bind to a Nectin-4 protein comprising the amino acid sequence of SEQ ID NO: 2 include antibodies that can bind to other Nectin-4-related proteins. For example, an antibody that binds to a Nectin-4 protein comprising the amino acid sequence of SEQ ID NO: 2 can bind to Nectin-4-related proteins such as Nectin-4 variants and their homologs or analogs.

[0283] In some embodiments, the anti-Nectin-4 antibodies provided herein are monoclonal antibodies.

[0284] In some embodiments, the antigen-binding fragment is a Fab, F(ab□)2, Fv, or scFv. In some embodiments, the antibody is a fully human antibody. In some embodiments, the antibody is an IgG1 and the light chain is a kappa light chain. In some embodiments, the antibody or antigen-binding fragment thereof is recombinantly produced.

[0285] In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:4 (cDNA sequence of SEQ ID NO:3) and / or a light chain comprising the amino acid sequence of SEQ ID NO:6 (cDNA sequence of SEQ ID NO:5), as shown in Figures 1B and 1C.

[0286] In some embodiments, the anti-Nectin-4 antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising a CDR having the amino acid sequence of the complementarity-determining region (CDR) of the heavy chain variable region set forth in SEQ ID NO: 22 (which is the amino acid sequence ranging from the 20th amino acid (glutamic acid) to the 136th amino acid (serine) of SEQ ID NO: 7), and a light chain variable region comprising a CDR having the amino acid sequence of the CDR of the light chain variable region set forth in SEQ ID NO: 23 (which is the amino acid sequence ranging from the 23rd amino acid (aspartic acid) to the 130th amino acid (arginine) of SEQ ID NO: 8). In certain embodiments, the anti-Nectin-4 antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising complementarity-determining region 1 (CDR-H1), CDR-H2, and CDR-H3, which contain the corresponding amino acid sequences of CDR-H1, CDR-H2, and CDR-H3 in the heavy chain variable region sequence shown in SEQ ID NO: 22 (which is the amino acid sequence ranging from the 20th amino acid (glutamic acid) to the 136th amino acid (serine) of SEQ ID NO: 7), and a light chain variable region comprising CDR-L1, CDR-L2, and CDR-L3, which contain the corresponding amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 in the light chain variable region sequence shown in SEQ ID NO: 23 (which is the amino acid sequence ranging from the 23rd amino acid (aspartic acid) to the 130th amino acid (arginine) of SEQ ID NO: 8). In some embodiments, the anti-Nectin-4 antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising a CDR consisting of the amino acid sequence of the complementarity-determining region (CDR) of the heavy chain variable region set forth in SEQ ID NO: 22 (which is the amino acid sequence ranging from the 20th amino acid (glutamic acid) to the 136th amino acid (serine) of SEQ ID NO: 7), and a light chain variable region comprising a CDR consisting of the amino acid sequence of the CDR of the light chain variable region set forth in SEQ ID NO: 23 (which is the amino acid sequence ranging from the 23rd amino acid (aspartic acid) to the 130th amino acid (arginine) of SEQ ID NO: 8).In certain embodiments, the anti-Nectin-4 antibody or antigen-binding fragment thereof comprises a heavy chain variable region including complementarity-determining region 1 (CDR-H1), CDR-H2, and CDR-H3 consisting of the corresponding amino acid sequences of CDR-H1, CDR-H2, and CDR-H3 in the heavy chain variable region sequence shown in SEQ ID NO: 22 (which is the amino acid sequence ranging from the 20th amino acid (glutamic acid) to the 136th amino acid (serine) of SEQ ID NO: 7), and a light chain variable region including CDR-L1, CDR-L2, and CDR-L3 consisting of the corresponding amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 in the light chain variable region sequence shown in SEQ ID NO: 23 (which is the amino acid sequence ranging from the 23rd amino acid (aspartic acid) to the 130th amino acid (arginine) of SEQ ID NO: 8). SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 7, and SEQ ID NO: 8 are shown in Figures 1D and 1E and are listed below. TIFF2025501978000008.tif164159

[0287] CDR sequences can be determined according to well-known numbering systems. As mentioned above, CDR regions are well known to those skilled in the art and are defined by well-known numbering systems. For example, Kabat complementarity-determining regions (CDRs) are based on sequence variability and are the most commonly used (see, for example, Kabat et al., supra). Instead, Chothia refers to the position of the structural loop (see, for example, Chothia and Lesk, 1987, J. Mol. Biol. 196:901-17). When numbered using Kabat's numbering rules, the end of the Chothia CDR-H1 loop varies between H32 and H34 depending on the length of the loop (this is because the Kabat numbering scheme places insertions at H35A and H35B; if neither 35A nor 35B is present, the loop ends at 32; if only 35A is present, the loop ends at 33; if both 35A and 35B are present, the loop ends at 34). AbM hypervariable regions represent a compromise between Kabat CDRs and Chothia structural loops and are used by Oxford Molecular's AbM antibody modeling software (see, e.g., Antibody Engineering Vol. 2 (Kontermann and Dubel eds., 2nd ed. 2010)). The "contact" hypervariable regions are based on the analysis of available complex crystal structures. Another universal numbering system that has been developed and widely adopted is the ImMunoGeneTics (IMGT) Information System® (Lafranc et al., 2003, Dev. Comp. Immunol. 27(1):55-77). IMGT is an integrated information system specialized for immunoglobulins (IGs), T cell receptors (TCRs), and major histocompatibility complexes (MHCs) of humans and other vertebrates. Herein, CDRs are referred to both with respect to amino acid sequence and location within the light or heavy chain. Because the "locations" of CDRs within the structure of immunoglobulin variable domains are conserved across species and reside within structures called loops, CDR and framework residues are readily identified by using a numbering system that aligns variable domain sequences according to structural features.This information can be used in grafting and substituting CDR residues from one species' immunoglobulin into an acceptor framework, typically from a human antibody. An additional numbering system (AHon) has been developed by Honegger and Pluckthun, 2001, J. Mol. Biol. 309:657-70. For example, the correspondence between numbering systems, including the Kabat numbering and the IMGT-specific numbering system, is well known to those of skill in the art (see, e.g., Kabat, supra; Chothia and Lesk, supra; Martin, supra; Lefranc et al., supra). Residues from each of these hypervariable regions or CDRs are set forth in Table 1 above.

[0288] In some embodiments, the anti-Nectin-4 antibody or antigen-binding fragment thereof comprises a heavy chain variable region including CDRs (CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3) comprising the amino acid sequences of the CDRs of the heavy chain variable region shown in SEQ ID NO: 22 according to the Kabat numbering, and a light chain variable region including CDRs comprising the amino acid sequences of the CDRs of the light chain variable region shown in SEQ ID NO: 23 according to the Kabat numbering.

[0289] In some embodiments, the anti-Nectin-4 antibody or antigen-binding fragment thereof comprises a heavy chain variable region including CDRs (CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3) comprising the amino acid sequences of the CDRs of the heavy chain variable region shown in SEQ ID NO: 22 according to AbM numbering, and a light chain variable region including CDRs comprising the amino acid sequences of the CDRs of the light chain variable region shown in SEQ ID NO: 23 according to AbM numbering.

[0290] In other embodiments, the anti-Nectin-4 antibody or antigen-binding fragment thereof comprises a heavy chain variable region including CDRs (CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3) comprising the amino acid sequences of the CDRs of the heavy chain variable region shown in SEQ ID NO: 22 according to Chothia numbering, and a light chain variable region including CDRs comprising the amino acid sequences of the CDRs of the light chain variable region shown in SEQ ID NO: 23 according to Chothia numbering.

[0291] In other embodiments, the anti-Nectin-4 antibody or antigen-binding fragment thereof comprises a heavy chain variable region including CDRs (CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3) comprising the amino acid sequences of the CDRs of the heavy chain variable region shown in SEQ ID NO: 22 by Contact numbering, and a light chain variable region including CDRs comprising the amino acid sequences of the CDRs of the light chain variable region shown in SEQ ID NO: 23 by Contact numbering.

[0292] In still other embodiments, the anti-Nectin-4 antibody or antigen-binding fragment thereof comprises a heavy chain variable region including CDRs (CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3) comprising the amino acid sequences of the CDRs of the heavy chain variable region shown in SEQ ID NO: 22 according to the IMGT numbering system, and a light chain variable region including CDRs comprising the amino acid sequences of the CDRs of the light chain variable region shown in SEQ ID NO: 23 according to the IMGT numbering system.

[0293] In some embodiments, the anti-Nectin-4 antibody or antigen-binding fragment thereof comprises a heavy chain variable region including CDRs (CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3) consisting of the amino acid sequences of the CDRs of the heavy chain variable region shown in SEQ ID NO: 22 according to the Kabat numbering, and a light chain variable region including CDRs consisting of the amino acid sequences of the CDRs of the light chain variable region shown in SEQ ID NO: 23 according to the Kabat numbering.

[0294] In some embodiments, the anti-Nectin-4 antibody or antigen-binding fragment thereof comprises a heavy chain variable region including CDRs (CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3) consisting of the amino acid sequences of the CDRs of the heavy chain variable region shown in SEQ ID NO: 22 according to AbM numbering, and a light chain variable region including CDRs consisting of the amino acid sequences of the CDRs of the light chain variable region shown in SEQ ID NO: 23 according to AbM numbering.

[0295] In another embodiment, the anti-Nectin-4 antibody or antigen-binding fragment thereof comprises a heavy chain variable region including CDRs (CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3) consisting of the amino acid sequences of the CDRs of the heavy chain variable region shown in SEQ ID NO: 22 according to Chothia numbering, and a light chain variable region including CDRs consisting of the amino acid sequences of the CDRs of the light chain variable region shown in SEQ ID NO: 23 according to Chothia numbering.

[0296] In other embodiments, the anti-Nectin-4 antibody or antigen-binding fragment thereof comprises a heavy chain variable region including CDRs (CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3) consisting of the amino acid sequences of the CDRs of the heavy chain variable region shown in SEQ ID NO: 22 by Contact numbering, and a light chain variable region including CDRs consisting of the amino acid sequences of the CDRs of the light chain variable region shown in SEQ ID NO: 23 by Contact numbering.

[0297] In still another embodiment, the anti-Nectin-4 antibody or antigen-binding fragment thereof comprises a heavy chain variable region including CDRs (CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3) consisting of the amino acid sequences of the CDRs of the heavy chain variable region shown in SEQ ID NO: 22 according to the IMGT numbering system, and a light chain variable region including CDRs consisting of the amino acid sequences of the CDRs of the light chain variable region shown in SEQ ID NO: 23 according to the IMGT numbering system.

[0298] As noted above, CDR sequences according to different numbering systems can be readily determined using online tools such as those provided by the Antigen Receptor Numbering and Receptor Classification (ANARCI). For example, the heavy chain CDR sequences in SEQ ID NO: 22 and the light chain CDR sequences in SEQ ID NO: 23 according to the Kabat numbering as determined by ANARCI are listed in Table 4 below.

[0299] (Table 4) TIFF2025501978000009.tif30160

[0300] As another example, the heavy chain CDR sequences in SEQ ID NO: 22 and the light chain CDR sequences in SEQ ID NO: 23 according to the IMGT numbering as determined by ANARCI are listed in Table 5 below.

[0301] (Table 5) TIFF2025501978000010.tif30160

[0302] In some embodiments, the antibody or antigen-binding fragment thereof comprises CDR-H1 comprising the amino acid sequence of SEQ ID NO: 9, CDR-H2 comprising the amino acid sequence of SEQ ID NO: 10, CDR-H3 comprising the amino acid sequence of SEQ ID NO: 11, CDR-L1 comprising the amino acid sequence of SEQ ID NO: 12, CDR-L2 comprising the amino acid sequence of SEQ ID NO: 13, and CDR-L3 comprising the amino acid sequence of SEQ ID NO: 14.

[0303] In some embodiments, the antibody or antigen-binding fragment thereof comprises CDR-H1 comprising the amino acid sequence of SEQ ID NO: 16, CDR-H2 comprising the amino acid sequence of SEQ ID NO: 17, CDR-H3 comprising the amino acid sequence of SEQ ID NO: 18, CDR-L1 comprising the amino acid sequence of SEQ ID NO: 19, CDR-L2 comprising the amino acid sequence of SEQ ID NO: 20, and CDR-L3 comprising the amino acid sequence of SEQ ID NO: 21.

[0304] In some embodiments, the antibody or antigen-binding fragment thereof comprises CDR-H1 consisting of the amino acid sequence of SEQ ID NO: 9, CDR-H2 consisting of the amino acid sequence of SEQ ID NO: 10, CDR-H3 consisting of the amino acid sequence of SEQ ID NO: 11, CDR-L1 consisting of the amino acid sequence of SEQ ID NO: 12, CDR-L2 consisting of the amino acid sequence of SEQ ID NO: 13, and CDR-L3 consisting of the amino acid sequence of SEQ ID NO: 14.

[0305] In some embodiments, the antibody or antigen-binding fragment thereof comprises CDR-H1 consisting of the amino acid sequence of SEQ ID NO: 16, CDR-H2 consisting of the amino acid sequence of SEQ ID NO: 17, CDR-H3 consisting of the amino acid sequence of SEQ ID NO: 18, CDR-L1 consisting of the amino acid sequence of SEQ ID NO: 19, CDR-L2 consisting of the amino acid sequence of SEQ ID NO: 20, and CDR-L3 consisting of the amino acid sequence of SEQ ID NO: 21.

[0306] In some embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:22 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:23.

[0307] In some embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region consisting of the amino acid sequence of SEQ ID NO:22 and a light chain variable region consisting of the amino acid sequence of SEQ ID NO:23.

[0308] In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence ranging from amino acid 20 (glutamic acid) to amino acid 466 (lysine) of SEQ ID NO:7, and a light chain comprising the amino acid sequence ranging from amino acid 23 (aspartic acid) to amino acid 236 (cysteine) of SEQ ID NO:8.

[0309] In some embodiments, the antibody comprises a heavy chain consisting of the amino acid sequence ranging from amino acid 20 (glutamic acid) to amino acid 466 (lysine) of SEQ ID NO:7, and a light chain consisting of the amino acid sequence ranging from amino acid 23 (aspartic acid) to amino acid 236 (cysteine) of SEQ ID NO:8.

[0310] In some embodiments, amino acid sequence modification(s) of the antibodies described herein are contemplated. For example, it may be desirable to optimize the binding affinity and / or other biological properties of the antibody, including, but not limited to, specificity, thermal stability, expression level, effector function, glycosylation, reduced immunogenicity, or solubility. Thus, in addition to the antibodies described herein, it is contemplated that antibody variants can be prepared. For example, antibody variants can be prepared by introducing appropriate nucleotide changes into the encoding DNA and / or by synthesis of the desired antibody or polypeptide. Those skilled in the art will recognize that amino acid changes can alter post-translational processing of the antibody, for example, by changing the number or position of glycosylation sites or altering membrane anchoring properties.

[0311] In some embodiments, the antibodies provided herein are chemically modified, for example, by the covalent attachment of any type of molecule to the antibody. Antibody derivatives can include antibodies chemically modified by, for example, glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization with known protecting / blocking groups, proteolytic cleavage, conjugation to cellular ligands or other proteins, etc. Any of a number of chemical modifications can be performed by known techniques, including, but not limited to, specific chemical cleavage, acetylation, formulation, metabolic synthesis of tunicamycin, etc. In addition, the antibody can contain one or more non-classical amino acids.

[0312] Mutations can be substitutions, deletions, or insertions of one or more codons encoding a single domain antibody or polypeptide, resulting in a change in the amino acid sequence compared to the original antibody or polypeptide. Amino acid substitutions can be, for example, conservative amino acid substitutions, resulting from the replacement of one amino acid with another amino acid containing similar structural and / or chemical properties, such as the replacement of a leucine with a serine. Standard techniques known to those skilled in the art can be used to introduce mutations into the nucleotide sequences encoding the molecules provided herein, including, for example, site-directed mutagenesis and PCR-mediated mutagenesis, which result in amino acid substitutions. Insertions or deletions can range from about 1 to 5 amino acids. In certain embodiments, substitutions, deletions, or insertions comprise fewer than 25 amino acid substitutions, fewer than 20 amino acid substitutions, fewer than 15 amino acid substitutions, fewer than 10 amino acid substitutions, fewer than 5 amino acid substitutions, fewer than 4 amino acid substitutions, fewer than 3 amino acid substitutions, or fewer than 2 amino acid substitutions compared to the original molecule. In specific embodiments, substitutions are conservative amino acid substitutions made at one or more predicted non-essential amino acid residues. Tolerated mutations can be determined by systematically inserting, deleting, or substituting amino acids in a sequence and testing the resulting variants for activity exhibited by the parent antibody.

[0313] Amino acid sequence insertions include amino- and / or carboxyl-terminal fusions ranging in length from one residue to polypeptides containing multiple residues, as well as intrasequence insertions of single or multiple amino acid residues. An example of a terminal insertion is an antibody with an N-terminal methionyl residue.

[0314] Antibodies generated by conservative amino acid substitutions are included in the present disclosure. In conservative amino acid substitutions, an amino acid residue is replaced with an amino acid residue containing a side chain with a similar charge. As described above, families of amino acid residues containing side chains with similar charges have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine), and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Alternatively, mutations can be introduced randomly along all or part of the coding sequence, such as by saturation mutagenesis, and the resulting mutants can be screened for biological activity to identify mutants that retain activity. After mutagenesis, the encoded protein can be expressed, the activity of the protein can be determined, and conservative substitutions (e.g., within a group of amino acids with similar properties and / or side chains) can be made to maintain or not significantly change the properties.

[0315] Amino acids can be grouped according to similarities in the properties of their side chains (see, e.g., Lehninger, Biochemistry 73-75 (2d ed. 1975)): (1) nonpolar: Ala (A), Val (V), Leu (L), Ile (I), Pro (P), Phe (F), Trp (W), Met (M); (2) uncharged polar: Gly (G), Ser (S), Thr (T), Cys (C), Tyr (Y), Asn (N), Gln (Q); (3) acidic: Asp (D), Glu (E), and (4) basic: Lys (K), Arg (R), His (H). Alternatively, naturally occurring residues can be divided into groups based on common side chain properties: (1) hydrophobic: norleucine, Met, Ala, Val, Leu, Ile; (2) neutral hydrophilic: Cys, Ser, Thr, Asn, Gln; (3) acidic: Asp, Glu; (4) basic: His, Lys, Arg; (5) residues that influence chain orientation: Gly, Pro; and (6) aromatic: Trp, Tyr, Phe.

[0316] For example, any cysteine ​​residue not involved in maintaining the proper conformation of the antibody may be substituted with another amino acid, e.g., alanine or serine, to improve the oxidative stability of the molecule and prevent aberrant crosslinking.

[0317] Modifications can be made using methods known in the art, such as oligonucleotide-mediated (site-directed) mutagenesis, alanine scanning, and PCR mutagenesis. Site-directed mutagenesis (see, e.g., Carter, 1986, Biochem J. 237:1-7, and Zoller et al., 1982, Nucl. Acids Res. 10:6487-500), cassette mutagenesis (see, e.g., Wells et al., 1985, Gene 34:315-23), or other known techniques can be performed on cloned DNA to produce anti-anti-MSLN antibody variant DNA.

[0318] Covalent modifications of antibodies are included within the scope of the present disclosure. Covalent modifications include reacting targeted amino acid residues of the antibody with organic derivatizing agents capable of reacting with selected side chains or the N- or C-terminal residues of the antibody. Other modifications include deamidation of glutaminyl and asparaginyl residues to the corresponding glutamyl and aspartyl residues, respectively, hydroxylation of proline and lysine, phosphorylation of the hydroxyl group of seryl or threonyl residues, methylation of the α-amino groups of lysine, arginine, and histidine side chains (see, e.g., Creighton, Proteins: Structure and Molecular Properties 79-86 (1983)), acetylation of the N-terminal amine, and amidation of any C-terminal carboxyl group.

[0319] Other types of covalent modifications of antibodies within the scope of this disclosure include altering the native glycosylation pattern of the antibody or polypeptide (see, e.g., Beck et al., 2008, Curr. Pharm. Biotechnol. 9:482-501, and Walsh, 2010, Drug Discov. Today 15:773-80), and linking the antibody to one of a variety of nonproteinaceous polymers, e.g., polyethylene glycol (PEG), polypropylene glycol, or polyoxyalkylenes, in the manner described, for example, in U.S. Pat. Nos. 4,640,835, 4,496,689, 4,301,144, 4,670,417, 4,791,192, or 4,179,337.

[0320] In certain embodiments, the antibodies or antigen-binding fragments provided herein comprise a heavy chain that has a particular homology or identity to the heavy chain set forth in SEQ ID NO:7 and a light chain that has a particular homology or identity to the light chain set forth in SEQ ID NO:8. Embodiments of heavy / light chains with such homology or identity are further provided below. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a heavy chain that has greater than 70% homology or identity to the heavy chain set forth in SEQ ID NO:7. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a heavy chain that has greater than 75% homology or identity to the heavy chain set forth in SEQ ID NO:7. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a heavy chain that has greater than 80% homology or identity to the heavy chain set forth in SEQ ID NO:7. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a heavy chain that has greater than 85% homology or identity to the heavy chain set forth in SEQ ID NO:7. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a heavy chain having greater than 90% homology or identity to the heavy chain set forth in SEQ ID NO: 7. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a heavy chain having greater than 95% homology or identity to the heavy chain set forth in SEQ ID NO: 7. In certain embodiments, the antibodies or antigen-binding fragments provided herein comprise a heavy chain having any of the provided homology or identity to the heavy chain set forth in SEQ ID NO: 7, and its CDRs (CDR-H1, CDR-H2, and CDR-H3) are identical to the CDRs in the heavy chain set forth in SEQ ID NO: 7. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a light chain having greater than 70% homology or identity to the light chain set forth in SEQ ID NO: 8. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a light chain having greater than 75% homology or identity to the light chain set forth in SEQ ID NO: 8. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a light chain having greater than 80% homology or identity to the light chain set forth in SEQ ID NO:8.In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a light chain having greater than 85% homology or identity to the light chain set forth in SEQ ID NO: 8. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a light chain having greater than 90% homology or identity to the light chain set forth in SEQ ID NO: 8. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a light chain having greater than 95% homology or identity to the light chain set forth in SEQ ID NO: 8. In certain embodiments, the antibodies or antigen-binding fragments provided herein comprise a light chain having any of the provided homology or identity to the light chain set forth in SEQ ID NO: 8, and whose CDRs (CDR-L1, CDR-L2, and CDR-L3) are identical to the CDRs in the light chain set forth in SEQ ID NO: 8. In certain embodiments, the antibodies or antigen-binding fragments provided herein comprise any of the homologous light chains and any of the homologous heavy chains provided in this paragraph, in any combination or permutation.

[0321] In certain embodiments, the antibodies or antigen-binding fragments provided herein comprise a heavy chain variable region having particular homology or identity to the heavy chain variable region set forth in SEQ ID NO: 22 and a light chain variable region having particular homology or identity to the light chain variable region set forth in SEQ ID NO: 23. Embodiments of heavy chain variable regions and light chain variable regions having such homology or identity are further provided below. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a heavy chain variable region having greater than 70% homology or identity to the heavy chain variable region set forth in SEQ ID NO: 22. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a heavy chain variable region having greater than 75% homology or identity to the heavy chain variable region set forth in SEQ ID NO: 22. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a heavy chain variable region having greater than 80% homology or identity to the heavy chain variable region set forth in SEQ ID NO: 22. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a heavy chain variable region having greater than 85% homology or identity to the heavy chain variable region set forth in SEQ ID NO: 22. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a heavy chain variable region having greater than 90% homology or identity to the heavy chain variable region set forth in SEQ ID NO: 22. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a heavy chain variable region having greater than 95% homology or identity to the heavy chain variable region set forth in SEQ ID NO: 22. In certain embodiments, the antibodies or antigen-binding fragments provided herein comprise a heavy chain variable region having any of the provided homologies or identities to the heavy chain variable region set forth in SEQ ID NO: 22, and the CDRs (CDR-H1, CDR-H2, and CDR-H3) are identical to the CDRs in the heavy chain variable region set forth in SEQ ID NO: 22. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a light chain variable region having greater than 70% homology or identity to the light chain variable region set forth in SEQ ID NO: 23. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a light chain variable region having greater than 75% homology or identity to the light chain variable region set forth in SEQ ID NO:23.In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a light chain variable region having greater than 80% homology or identity to the light chain variable region set forth in SEQ ID NO: 23. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a light chain variable region having greater than 85% homology or identity to the light chain variable region set forth in SEQ ID NO: 23. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a light chain variable region having greater than 90% homology or identity to the light chain variable region set forth in SEQ ID NO: 23. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a light chain variable region having greater than 95% homology or identity to the light chain variable region set forth in SEQ ID NO: 23. In certain embodiments, the antibodies or antigen-binding fragments provided herein comprise a light chain variable region having any of the provided homology or identity to the light chain variable region set forth in SEQ ID NO: 23, and the CDRs (CDR-L1, CDR-L2, and CDR-L3) are identical to the CDRs in the light chain variable region set forth in SEQ ID NO: 23. In certain embodiments, the antibodies or antigen-binding fragments provided herein comprise any of the homologous light chain variable regions and any of the homologous heavy chain variable regions provided in this paragraph, in any combination or permutation.

[0322] In some embodiments, the anti-Nectin-4 antibody provided herein comprises the heavy and light chain CDR regions of the antibody designated Ha22-2(2,4)6.1 produced by the hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267, or heavy and light chain CDR regions comprising amino acid sequences homologous to the amino acid sequences of the heavy and light chain CDR regions of Ha22-2(2,4)6.1, and the antibody retains desirable functional properties of the anti-Nectin-4 antibody designated Ha22-2(2,4)6.1 produced by the hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267.

[0323] In some embodiments, the anti-Nectin-4 antibody provided herein comprises the heavy and light chain CDR regions (CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3) of the antibody designated Ha22-2(2,4)6.1 produced by the hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267, or heavy and light chain CDR regions consisting of amino acid sequences homologous to the amino acid sequences of the heavy and light chain CDR regions of Ha22-2(2,4)6.1, wherein the antibody retains desirable functional properties of the anti-Nectin-4 antibody designated Ha22-2(2,4)6.1 produced by the hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267.

[0324] In some embodiments, the antibodies or antigen-binding fragments thereof provided herein comprise a humanized heavy chain variable region and a humanized light chain variable region; (a) the heavy chain variable region comprises CDRs (CDR-H1, CDR-H2, and CDR-H3) comprising the amino acid sequences of the heavy chain variable region CDRs shown in the antibody produced by the hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267; (b) The light chain variable region comprises CDRs (CDR-L1, CDR-L2, and CDR-L3) comprising the amino acid sequences of the light chain variable region CDRs shown in the antibody produced by the hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267.

[0325] In some embodiments, the antibodies or antigen-binding fragments thereof provided herein comprise a humanized heavy chain variable region and a humanized light chain variable region; (a) the heavy chain variable region comprises CDRs (CDR-H1, CDR-H2, and CDR-H3) consisting of the amino acid sequences of the heavy chain variable region CDRs shown in the antibody produced by the hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267; (b) The light chain variable region contains CDRs (CDR-L1, CDR-L2, and CDR-L3) consisting of the amino acid sequences of the light chain variable region CDRs shown in the antibody produced by the hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267.

[0326] In some embodiments, the anti-Nectin-4 antibodies provided herein comprise the heavy and light chain variable regions of the antibody designated Ha22-2(2,4)6.1 produced by the hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267, or heavy and light chain variable regions comprising amino acid sequences homologous to the amino acid sequences of the heavy and light chain variable regions of Ha22-2(2,4)6.1, and the antibodies retain the desired functional properties of the anti-Nectin-4 antibodies provided herein. In some embodiments, the anti-Nectin-4 antibodies provided herein comprise the heavy and light chain variable regions of the antibody designated Ha22-2(2,4)6.1, produced by the hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267, or heavy and light chain variable regions consisting of amino acid sequences homologous to the amino acid sequences of the heavy and light chain variable regions of Ha22-2(2,4)6.1, and the antibodies retain the desired functional properties of the anti-Nectin-4 antibodies provided herein. The constant regions of any subclass can be selected for the constant regions of the antibodies of the present disclosure. In one embodiment, a human IgG1 constant region can be used as the heavy chain constant region, and a human Ig kappa constant region can be used as the light chain constant region.

[0327] In some embodiments, the anti-Nectin-4 antibodies provided herein comprise the heavy and light chains of the antibody designated Ha22-2(2,4)6.1 produced by the hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267, or heavy and light chains comprising amino acid sequences homologous to the amino acid sequences of the heavy and light chains of Ha22-2(2,4)6.1, and the antibodies retain the desired functional properties of the anti-Nectin-4 antibodies provided herein. In some embodiments, the anti-Nectin-4 antibodies provided herein comprise the heavy and light chains of the antibody designated Ha22-2(2,4)6.1 produced by the hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267, or heavy and light chains consisting of amino acid sequences homologous to the amino acid sequences of the heavy and light chains of Ha22-2(2,4)6.1, and the antibodies retain the desired functional properties of the anti-Nectin-4 antibodies provided herein.

[0328] In some embodiments, the antibodies or antigen-binding fragments thereof provided herein comprise a heavy chain variable region and a light chain variable region, (a) the heavy chain variable region comprises an amino acid sequence that is at least 80% homologous or identical to the heavy chain variable region amino acid sequence of the antibody produced by the hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267; (b) the light chain variable region comprises an amino acid sequence that is at least 80% homologous or identical to the light chain variable region amino acid sequence of the antibody produced by the hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267.

[0329] In certain embodiments, the antibodies or antigen-binding fragments provided herein comprise a heavy chain variable region having specific homology or identity to the heavy chain variable region amino acid sequence of an antibody produced by a hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267, and a light chain variable region having specific homology or identity to the light chain variable region amino acid sequence of an antibody produced by a hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267. Embodiments of heavy chain and light chain variable regions having such homology or identity are provided further below. In some embodiments, the heavy chain variable region comprises an amino acid sequence that is at least 85% homologous or identical to the heavy chain variable region amino acid sequence of an antibody produced by a hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267. In other embodiments, the heavy chain variable region comprises an amino acid sequence that is at least 90% homologous or identical to the heavy chain variable region amino acid sequence of the antibody produced by the hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267. In yet other embodiments, the heavy chain variable region comprises an amino acid sequence that is at least 95% homologous or identical to the heavy chain variable region amino acid sequence of the antibody produced by the hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267. In other embodiments, the heavy chain variable region may be 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% homologous or identical to the heavy chain variable region amino acid sequence of the antibody produced by the hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267.In some embodiments, the light chain variable region comprises an amino acid sequence that is at least 85% homologous or identical to the light chain variable region amino acid sequence of an antibody produced by a hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267. In other embodiments, the light chain variable region comprises an amino acid sequence that is at least 90% homologous or identical to the light chain variable region amino acid sequence of an antibody produced by a hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267. In yet other embodiments, the light chain variable region comprises an amino acid sequence that is at least 95% homologous or identical to the light chain variable region amino acid sequence of an antibody produced by a hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267. In other embodiments, the light chain variable region can be 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% homologous or identical to the light chain variable region amino acid sequence of the antibody produced by the hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA- 11267. In certain embodiments, the antibodies or antigen-binding fragments provided herein comprise any homologous light chain variable region and any homologous heavy chain variable region provided in this paragraph, in any combination or permutation.

[0330] In other embodiments, the antibodies or antigen-binding fragments thereof provided herein comprise a heavy chain and a light chain: (a) the heavy chain comprises an amino acid sequence that is at least 80% homologous or identical to the heavy chain amino acid sequence of the antibody produced by the hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267; (b) the light chain comprises an amino acid sequence that is at least 80% homologous or identical to the light chain amino acid sequence of an antibody produced by the hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267.

[0331] In certain embodiments, the antibodies or antigen-binding fragments provided herein comprise a heavy chain having specific homology or identity to the heavy chain amino acid sequence of the antibody produced by the hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267, and a light chain having specific homology or identity to the light chain amino acid sequence of the antibody produced by the hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267. Embodiments of heavy and light chains having such homology or identity are provided further below. In some embodiments, the heavy chain comprises an amino acid sequence that is at least 85% homologous or identical to the heavy chain amino acid sequence of the antibody produced by the hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267. In other embodiments, the heavy chain comprises an amino acid sequence that is at least 90% homologous or identical to the heavy chain amino acid sequence of the antibody produced by the hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267. In yet other embodiments, the heavy chain comprises an amino acid sequence that is at least 95% homologous or identical to the heavy chain amino acid sequence of the antibody produced by the hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267. In other embodiments, the heavy chain may be 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% homologous or identical to the heavy chain amino acid sequence of the antibody produced by the hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267.In some embodiments, the light chain comprises an amino acid sequence that is at least 85% homologous or identical to the light chain amino acid sequence of an antibody produced by a hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267. In other embodiments, the light chain comprises an amino acid sequence that is at least 90% homologous or identical to the light chain amino acid sequence of an antibody produced by a hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267. In yet other embodiments, the light chain comprises an amino acid sequence that is at least 95% homologous or identical to the light chain amino acid sequence of an antibody produced by a hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA-11267. In other embodiments, the light chain can be 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% homologous or identical to the light chain amino acid sequence of the antibody produced by the hybridoma deposited under American Type Culture Collection (ATCC) Accession No. PTA- 11267. In certain embodiments, the antibodies or antigen-binding fragments provided herein comprise any homologous light chain and any homologous heavy chain provided in this paragraph, in any combination or permutation.

[0332] In some embodiments, the antibodies or antigen-binding fragments provided herein bind to a specific epitope of 191P4D12. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to the VC1 domain of 191P4D12. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to the VC1 domain but not the C1C2 domain of 191P4D12. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to amino acid residues 1 to 147 of 191P4D12. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to an epitope located at amino acid residues 1 to 147 of 191P4D12. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to amino acid residues 1 to 10 of 191P4D12. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to amino acid residues 11 to 20 of 191P4D12. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to amino acid residues 21 to 30 of 191P4D12. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to amino acid residues 31 to 40 of 191P4D12. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to amino acid residues 41 to 50 of 191P4D12. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to amino acid residues 51 to 60 of 191P4D12. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to amino acid residues 61 to 70 of 191P4D12. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to amino acid residues 71 to 80 of 191P4D12. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to amino acid residues 81 to 90 of 191P4D12.In some embodiments, the antibodies or antigen-binding fragments provided herein bind to amino acid residues 91 to 100 of 191P4D12. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to amino acid residues 101 to 110 of 191P4D12. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to amino acid residues 111 to 120 of 191P4D12. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to amino acid residues 121 to 130 of 191P4D12. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to amino acid residues 131 to 140 of 191P4D12. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to amino acid residues 141 to 147 of 191P4D12. The binding epitopes of certain embodiments of the antibodies or antigen-binding fragments thereof provided herein are determined and described in WO2012 / 047724, which is incorporated herein by reference in its entirety.

[0333] In some embodiments, the antibodies or antigen-binding fragments provided herein bind to an epitope of 191P4D12 that is shared among 191P4D12 variants observed in humans. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to an epitope of 191P4D12 that is shared among 191P4D12 polymorphisms observed in humans. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to an epitope of 191P4D12 that is shared among 191P4D12 polymorphisms observed in human cancer. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to an epitope of 191P4D12 that will bind, internalize, disrupt, or modulate the biological function of 191P4D12 or a 191P4D12 variant. In some embodiments, the antibodies or antigen-binding fragments thereof provided herein bind to an epitope of 191P4D12 that will disrupt interactions between 191P4D12 and ligands, substrates, and binding partners.

[0334] The engineered antibodies provided herein include those in which modifications have been made to framework residues within the VH and / or VL (e.g., to improve the properties of the antibody). Typically, such framework modifications are made to reduce the immunogenicity of the antibody. For example, one approach is to "backmutate" one or more framework residues to the corresponding germline sequence. More specifically, antibodies that have undergone somatic mutation may contain framework residues that differ from the germline sequence from which the antibody is derived. Such residues can be identified by comparing the antibody framework sequence to the germline sequence from which the antibody is derived. To return framework region sequences to their germline configuration, somatic mutations can be "backmutated" to germline sequences (e.g., "backmutating" leucine to methionine),...

Claims

1. 1. A medicament for use in a method of treating muscle-invasive urothelial carcinoma (MIUC) or muscle-invasive bladder cancer (MIBC) in a human subject, comprising an effective amount of an antibody drug conjugate (ADC), The method comprises: (a) administering to the subject the effective amount of an ADC; the ADC comprises an antibody or antigen-binding fragment thereof that binds to 191P4D12 (Nectin-4) conjugated to one or more units of monomethyl auristatin E (MMAE); the antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising a CDR comprising the amino acid sequence of the complementarity-determining region (CDR) of the heavy chain variable region set forth in SEQ ID NO: 22, and a light chain variable region comprising a CDR comprising the amino acid sequence of the CDR of the light chain variable region set forth in SEQ ID NO: 23; the subject is ineligible to receive cisplatin therapy (cisplatin-ineligible), and (i) a pathological complete response rate (pCRR) of at least 30%, or (ii) a pathological downstaging rate (pDSR) of at least 50%; The medicine.

2. The method of claim 1, wherein the method comprises administering three cycles of the ADC to the subject.

3. The pharmaceutical described in claim 1, wherein the method of treatment further comprises radical cystectomy and pelvic lymph node dissection (RC+PLND).

4. The pharmaceutical described in claim 3, wherein the method involves administering the ADC to the human subject, and the human subject receiving the RC+PLND approximately 4 to approximately 12 weeks after the ADC is administered to the human subject.

5. The pharmaceutical described in claim 1, wherein the method further comprises (b) performing radical cystectomy and pelvic lymph node dissection (RC+PLND) on the subject.

6. The pharmaceutical described in claim 5, wherein, in the method, (b) is performed approximately 4 to approximately 12 weeks after (a).

7. 1. A medicament for use in a method of treating muscle-invasive urothelial carcinoma (MIUC) or muscle-invasive bladder cancer (MIBC) in a human subject, comprising an effective amount of an antibody drug conjugate (ADC), the method comprises administering to the subject multiple cycles of the effective amount of the ADC; and (a) the ADC comprises an antibody or antigen-binding fragment thereof that binds to 191P4D12 (Nectin-4) conjugated to one or more units of monomethyl auristatin E (MMAE); the antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising a CDR comprising the amino acid sequence of the complementarity-determining region (CDR) of the heavy chain variable region set forth in SEQ ID NO: 22, and a light chain variable region comprising a CDR comprising the amino acid sequence of the CDR of the light chain variable region set forth in SEQ ID NO: 23; (b) the subject is ineligible to receive cisplatin therapy (cisplatin-ineligible), and (c) the number of cycles of ADC therapy administered to the cisplatin-ineligible subject is less than or equal to the number of cycles of standard of care (SOC) therapy used to treat cisplatin-eligible subjects with MIUC or MIBC; The medicine.

8. The SOC regimen for a cisplatin-eligible subject comprises: i. cisplatin, ii. methotrexate, vinblastine, doxorubicin, and cisplatin (MVAC); iii. gemcitabine and cisplatin; iv. administering a programmed cell death 1 (PD-1) inhibitor, or v. Administering a programmed cell death-ligand 1 (PD-L1) inhibitor The pharmaceutical composition of claim 7, comprising:

9. The method of claim 7, wherein the number of cycles of ADC therapy administered to the cisplatin-ineligible subject is 3 or 4.

10. 1. A medicament for use in a method of administering neoadjuvant or perioperative therapy for treating muscle-invasive urothelial carcinoma (MIUC) or muscle-invasive bladder cancer (MIBC) in a human subject, comprising an effective amount of an antibody drug conjugate (ADC), the method comprises administering to the subject multiple cycles of the effective amount of the ADC; and (a) the ADC comprises an antibody or antigen-binding fragment thereof that binds to 191P4D12 (Nectin-4) conjugated to one or more units of monomethyl auristatin E (MMAE); the antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising a CDR comprising the amino acid sequence of the complementarity-determining region (CDR) of the heavy chain variable region set forth in SEQ ID NO: 22, and a light chain variable region comprising a CDR comprising the amino acid sequence of the CDR of the light chain variable region set forth in SEQ ID NO: 23; (b) the subject is ineligible to receive cisplatin therapy (cisplatin-ineligible), and (c) the subject remains eligible for radical cystectomy and pelvic lymph node dissection (RC+PLND) surgery after said multiple cycles of treatment with the ADC. The medicine.

11. The pharmaceutical described in claim 10, wherein in the method, the ADC is administered as neoadjuvant therapy (i) before the surgery, or (ii) before and after the surgery.

12. 1. A medicament for use in a method of treating muscle-invasive urothelial carcinoma (MIUC) or muscle-invasive bladder cancer (MIBC) in a human subject, comprising an effective amount of an antibody drug conjugate (ADC), the method comprises administering to the subject multiple cycles of the effective amount of the ADC; and (a) the ADC comprises an antibody or antigen-binding fragment thereof that binds to 191P4D12 (Nectin-4) conjugated to one or more units of monomethyl auristatin E (MMAE); the antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising a CDR comprising the amino acid sequence of the complementarity-determining region (CDR) of the heavy chain variable region set forth in SEQ ID NO: 22, and a light chain variable region comprising a CDR comprising the amino acid sequence of the CDR of the light chain variable region set forth in SEQ ID NO: 23; (b) the subject is ineligible to receive cisplatin therapy (cisplatin-ineligible); (c) the efficacy of said treatment of said subject with said effective amount of said ADC is similar to the efficacy of treatment observed in cisplatin-eligible patients treated with a standard of care (SOC) regimen used to treat cisplatin-eligible subjects with MIUC or MIBC; The medicine.

13. The method of claim 12, wherein the efficacy of the treatment is at least as effective as the efficacy of the treatment observed in a SOC for cisplatin-eligible subjects.

14. The pharmaceutical composition of claim 12, wherein the measure of efficacy of treatment is one or more of pathological complete response rate (pCRR), pathological downstaging rate (pDSR), disease-free survival (DFS), recurrence-free survival (EFS), overall survival (OS), progression-free survival (PFS), and duration of response (DoR). (i) the pCRR for the cisplatin-ineligible subjects is at least 30%, or (ii) the pDSR for the cisplatin-ineligible subjects is at least 50%; The pharmaceutical composition according to claim 14.

16. The SOC therapy for the cisplatin-eligible subject comprises: (i) cisplatin, (ii) methotrexate, vinblastine, doxorubicin, and cisplatin (MVAC); (iii) gemcitabine and cisplatin; (iv) a programmed cell death 1 (PD-1) inhibitor, optionally wherein the PD-1 inhibitor is nivolumab or pembrolizumab, or (v) a programmed cell death-ligand 1 (PD-L1) inhibitor, optionally wherein the PD-L1 inhibitor is selected from the group consisting of atezolizumab, avelumab, and durvalumab; The pharmaceutical composition of claim 12, comprising:

17. (i) the antibody or antigen-binding fragment thereof comprises CDR-H1 comprising the amino acid sequence of SEQ ID NO:9, CDR-H2 comprising the amino acid sequence of SEQ ID NO:10, CDR-H3 comprising the amino acid sequence of SEQ ID NO:11, CDR-L1 comprising the amino acid sequence of SEQ ID NO:12, CDR-L2 comprising the amino acid sequence of SEQ ID NO:13, and CDR-L3 comprising the amino acid sequence of SEQ ID NO:14; or (ii) the antibody or antigen-binding fragment thereof comprises CDR-H1 comprising the amino acid sequence of SEQ ID NO: 16, CDR-H2 comprising the amino acid sequence of SEQ ID NO: 17, CDR-H3 comprising the amino acid sequence of SEQ ID NO: 18, CDR-L1 comprising the amino acid sequence of SEQ ID NO: 19, CDR-L2 comprising the amino acid sequence of SEQ ID NO: 20, and CDR-L3 comprising the amino acid sequence of SEQ ID NO: 21; The pharmaceutical composition according to any one of claims 1 to 16.

18. The pharmaceutical according to any one of claims 1 to 16, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 22 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:

23.

19. The pharmaceutical according to any one of claims 1 to 16, wherein the antibody comprises a heavy chain comprising an amino acid sequence ranging from the 20th amino acid (glutamic acid) to the 466th amino acid (lysine) of SEQ ID NO: 7, and a light chain comprising an amino acid sequence ranging from the 23rd amino acid (aspartic acid) to the 236th amino acid (cysteine) of SEQ ID NO:

8.

20. the antigen-binding fragment: (i) is a Fab, F(ab')2, Fv, or scFv; or (ii) recombinantly produced; The pharmaceutical composition according to any one of claims 1 to 16.

21. The antibody (i) Is it a fully human antibody? (ii) IgG1, and the light chain is a kappa light chain; or (iii) recombinantly produced; The pharmaceutical composition according to any one of claims 1 to 16.

22. the antibody or antigen-binding fragment is conjugated to each unit of MMAE via a linker, optionally the linker is an enzyme-cleavable linker, and the linker forms a bond with a sulfur atom of the antibody or antigen-binding fragment thereof; optionally the linker has the formula -Aa-Ww-Yy-, where -A- is a Stretcher unit, a is 0 or 1, -W- is an amino acid unit, w is an integer ranging from 0 to 12, -Y- is a Spacer unit, and y is 0, 1, or 2; optionally the Stretcher unit has the structure of formula (1) below, the amino acid unit is valine-citrulline, and the Spacer unit is a PAB group comprising the structure of formula (2) below: Optionally, the Stretcher unit forms a bond with a sulfur atom of the antibody or antigen-binding fragment thereof, and the Spacer unit is linked to MMAE via a carbamate group.

23. The ADC is (i) 1 to 20 units of MMAE per antibody or antigen-binding fragment thereof; (ii) 1 to 10 units of MMAE per antibody or antigen-binding fragment thereof; (iii) 2 to 8 units of MMAE per antibody or antigen-binding fragment thereof; or (iv) 3 to 5 units of MMAE per antibody or antigen-binding fragment thereof The pharmaceutical composition according to any one of claims 1 to 16, comprising:

24. The ADC has the following structure: and wherein L- represents the antibody or antigen-binding fragment thereof; and (i) p is 1 to 10, (ii) p is 2 to 8, (iii) p is 3 to 5, (iv) p is 3 to 4, (v) p is about 4, or (vi) the effective amount of said antibody drug conjugate has a mean p-value of about 3.8; The pharmaceutical composition according to any one of claims 1 to 16.

25. The ADC is (i) a pharmaceutical composition comprising L-histidine, polysorbate-20, and trehalose dihydrate; (ii) a pharmaceutical composition comprising about 20 mM L-histidine, about 0.02% (w / v) polysorbate-20, about 5.5% (w / v) trehalose dihydrate, and hydrochloride, wherein the pH of the pharmaceutical composition is about 6.0 at 25°C; or (iii) A pharmaceutical composition comprising about 9 mM histidine, about 11 mM histidine hydrochloride monohydrate, about 0.02% (w / v) polysorbate-20, and about 5.5% (w / v) trehalose dihydrate, wherein the pH of the pharmaceutical composition is about 6.0 at 25°C. The pharmaceutical composition according to any one of claims 1 to 16, which is formulated in

26. The ADC has the following structure: and wherein L- represents the antibody or antigen-binding fragment thereof; p is about 3 to about 4; the antibody comprises a heavy chain comprising the amino acid sequence ranging from the 20th amino acid (glutamic acid) to the 466th amino acid (lysine) of SEQ ID NO: 7 and a light chain comprising the amino acid sequence ranging from the 23rd amino acid (aspartic acid) to the 236th amino acid (cysteine) of SEQ ID NO: 8; the ADC is administered at a dose of about 1.25 mg / kg of subject body weight, and the dose is administered by IV infusion on days 1 and 8 of a 3-week cycle.

27. The method of any one of claims 1 to 16, wherein the subject has cT2-T4aN0M0 stage MIBC.

28. The subject meets the following criteria: (a) a GFR of less than 60 mL / min but greater than or equal to 30 mL / min, wherein the GFR is measured by the Cockcroft-Gault formula, the Modification of Diet in Renal Disease formula (MDRD), or a 24-hour urine collection; (b) ECOG performance status of 2; (c) hearing loss of grade 2 or greater on the NCI CTCAE version 4.03; and (d) NYHA Class III heart failure The method according to any one of claims 1 to 16, wherein the patient is considered ineligible for cisplatin if one or more of the following conditions are met:

29. The method of any one of claims 1 to 16, wherein the subject has not received previous systemic treatment, chemoradiotherapy, and / or radiotherapy for MIBC.

30. The medicament of any one of claims 1 to 16, wherein the subject is not receiving an immune checkpoint inhibitor (CPI), and optionally the CPI is a programmed cell death 1 (PD-1) inhibitor or a programmed cell death-ligand 1 (PD-L1) inhibitor.

31. The method of any one of claims 1 to 16, wherein the subject is not receiving a CD137 agonist, a CTLA-4 inhibitor, or an OX-40 agonist.

32. The medicament of any one of claims 1 to 16, wherein the effective amount of the ADC is about 1 to about 10 mg / kg, about 1 to about 5 mg / kg, about 1 to about 2.5 mg / kg, about 1 to about 1.25 mg / kg, about 0.25 mg / kg, about 0.5 mg / kg, about 0.75 mg / kg, about 1.0 mg / kg, about 1.25 mg / kg, about 1.5 mg / kg, about 1.75 mg / kg, about 2.0 mg / kg, about 2.25 mg / kg, or about 2.5 mg / kg of the subject's body weight.

33. The pharmaceutical composition according to any one of claims 1 to 16, wherein the effective amount of the ADC is about 1 mg / kg of subject body weight.

34. The pharmaceutical composition of any one of claims 1 to 16, wherein the effective amount of the ADC is about 1.25 mg / kg of subject body weight.

35. The pharmaceutical composition of claim 1, wherein in (a), the effective amount of the ADC is administered to the subject on days 1 and 8 of a cycle every three weeks for a total of three cycles.

36. 36. The method of claim 35, wherein the effective amount of the ADC is about 1.25 mg / kg of subject body weight.

37. (a), wherein the effective amount of the ADC is administered to the subject on days 1 and 8 of a three-week cycle for a total of three cycles; and the method further comprising: (b) administering to the subject the effective amount of the ADC on days 1 and 8 of every 3-week cycle for a total of 6 cycles, starting about 8 weeks after the subject receives the RC+PLND. The pharmaceutical composition according to claim 3.

38. 38. The method of claim 37, wherein the effective amount of the ADC is about 1.25 mg / kg of subject body weight.

39. (a), wherein the effective amount of the ADC is administered to the subject on days 1 and 8 of a three-week cycle for a total of three cycles; and the method further comprising: (c) about 8 weeks after (b), administering to the subject the effective amount of the ADC on days 1 and 8 of every 3-week cycle for a total of 3 cycles; The pharmaceutical composition according to claim 5.

40. A pharmaceutical described in any one of claims 7 to 16, wherein in the method, the effective amount of the ADC is administered to the human subject on days 1 and 8 of a three-week cycle for a total of three cycles.

41. The pharmaceutical described in any one of claims 1 to 16, wherein in the method, the ADC is administered by intravenous (IV) injection or infusion.

42. The pharmaceutical agent according to any one of claims 1 to 16, wherein the ADC is enfortumab vedotin (EV).

43. The method of any one of claims 1 to 16, wherein the overall survival of the subject is extended by at least 2 months, at least 4 months, at least 6 months, at least 8 months, at least 10 months, or at least 12 months.

44. A pharmaceutical described in any one of claims 1 to 16, wherein in the method, the ADC is administered as monotherapy.

Images