Methods of treating patients suffering from locally advanced or metastatic urethral epithelial cancer with antibody drug conjugates (ADCs) that bind to 191P4D12 protein in combination with palbolizumab

Abstract

Provided herein are methods of treating cancer with an antibody drug conjugate (ADC) that binds to the 191P4D12 protein (connexin-4) and palbolizumab.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of U.S. Application No. 63 / 392,067, filed on July 25, 2022, U.S. Application No. 63 / 402,830, filed on August 31, 2022, and U.S. Application No. 63 / 504,183, filed on May 24, 2023, the disclosures of each of which are incorporated herein by reference in their entirety.

[0003] Sequence Listing

[0004] This application contains a computer-readable sequence listing submitted with this application in XML file format, the entire contents of which are incorporated herein by reference in their entirety. The sequence listing XML file submitted with this application is titled "14369-295-228_SEQLISTING.xml", was created on July 18, 2023, and is 62,806 bytes in size. 1. Field of the Invention

[0005] Provided herein are methods for treating cancer using an antibody drug conjugate (ADC) that binds to the 191P4D12 protein (Nectin-4). Specifically, provided herein are methods for treating patients with unresectable locally advanced or metastatic urothelial carcinoma who are unable to receive cisplatin-based chemotherapy using an antibody drug conjugate (ADC) that binds to the 191P4D12 protein in combination with pembrolizumab. 2. Background Technology

[0006] Cancer is the leading cause of death in the United States for people aged 35 to 65, and it is the second leading cause of death in the world. It is estimated that in 2019, the United States will have approximately 1.7 million new cancer cases and approximately 610,000 deaths from cancer (National Cancer Institute.2019.Cancer Stat Facts:Cancer of Any Site.seer.cancer.gov / statfacts / html / all.html.Accessed on June 5, 2019). In 2018, it was estimated that there were 18.1 million new cancer cases and approximately 9.6 million deaths from cancer in 2018 (World Health Organization.Press Release.September 2018.who.int / cancer / PRGlobocanFinal.pdf.Accessed on June 5, 2019). Most deaths now occur in patients with metastatic cancer. In fact, over the past 20 years, advances in treatment, including surgery, radiotherapy, and adjuvant chemotherapy have cured most patients with localized cancer. Patients whose cancer presents or relapses as metastatic disease derive modest benefit from conventional therapies only in terms of overall survival (OS), and cure is rare.

[0007] New therapeutic strategies for advanced and / or metastatic cancer include targeting molecular pathways important for cancer cell survival and novel cytotoxic compounds. The benefits of these novel drugs are reflected in long-term survival; however, the majority of patients with distant metastases still have poor outcomes and new therapies are needed.

[0008] 191P4D12 (also known as connexin-4) is a 66 kDa type I transmembrane protein that belongs to the connexin family of adhesion molecules. It consists of an extracellular domain (ECD) containing three immunoglobulin (Ig)-like domains, a transmembrane helix, and an intracellular region (Takai et al., Annu Rev Cell Dev Biol (2008); 24: 309-42). Connexins are believed to mediate Ca binding via both homophilic and heterophilic trans-interactions at adherens junctions. 2+ Independent cell-cell adhesion, where it recruits cadherins and regulates cytoskeletal rearrangements (Rikitake et al., Cell Mol Life Sci (2008); 65(2): 253-63.). Connexin-4 has low sequence identity with other connexin family members, and in the ECD, the sequence identity ranges from 25% to 30% (Reymond et al., Biol Chem (2001); 276(46): 43205-15).

[0009] The three Ig-like domains in the ECD of connexin-4 are called V, C1, and C2. The C1 domain is responsible for cis interactions (homodimerization), while the V domain of most connexin molecules promotes 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.).

[0010] Connexin-4 was originally identified bioinformatically and cloned from human trachea (Reymond et al., J Biol Chem (2001) 276(46):43205-15). Using suppression subtractive hybridization on a pool of urothelial carcinoma specimens, connexin-4 was identified as significantly upregulated in urothelial carcinoma. Characterization of expression in multiple tumor specimens at the RNA level and by immunohistochemistry (IHC) also demonstrated elevated levels of connexin-4 in breast, pancreatic, lung, and other cancers (Challita-Eid et al., Cancer Res (2016); 76(10):3003-13).

[0011] Connexin-4 has been found to be expressed in a variety of cancers, particularly urothelial, breast, lung, pancreatic, and ovarian cancers, with higher expression levels being associated with disease progression and / or poor prognosis (Fabre-Lafay et al., BMC Cancer (2007); 7:73).

[0012] PD-1

[0013] PD-1 is recognized as an important molecule for immune regulation and maintenance of peripheral tolerance. PD-1 is moderately expressed on naive T, B, and NKT cells and is upregulated by T / B cell receptor signaling on lymphocytes, monocytes, and myeloid cells (Sharpe, Arlene H. et al., The function of programmed cell death 1 and its ligands inregulating autoimmunity and infection. Nature Immunology (2007); 8: 239-245).

[0014] Two known ligands of PD-1, PD-L1 (B7-H1) and PD-L2 (B7-DC), are expressed in human cancers arising in various tissues. In large cohorts of cancers such as ovarian cancer, renal cancer, colorectal cancer, pancreatic cancer, liver cancer, and melanoma, PD-L1 expression has been shown to be associated with a poor prognosis and shortened overall survival regardless of subsequent treatment (Dong, Haidong et al., Tumor-associated B7-H1 promotes T-cell apoptosis: a potential mechanism of immune evasion. Nat Med. 2002 Aug;8(8):793-800; Yang, Wanhua et al., PD-1 interaction contributes to the functional suppression of T-cell responses to human uveal melanoma cells in vitro. Invest Ophthalmol Vis Sci. 2008 Jun;49(6(2008):49:2518-2525; Ghebeh, Hazem et al., The B7-H1 (PD-L1) T lymphocyte-inhibitory molecule is expressed in breast cancer patients with infiltrating ductal carcinoma: correlation with important high-risk prognostic factors.Neoplasia (2006) 8: 190-198; Hamanishi, Junzo et al., Programmed cell death 1 ligand 1 andtumor-infiltrating CD8+ T lymphocytes are prognostic factors of human ovariancancer. Proc. Natl. Acad. Sci. USA (2007): 104: 3360-3365; Thompson, R Houston and Eugene D Kwon, Significance of B7-H1 overexpression in kidney cancer.C“nical genitourinCancer(2006):5:206 - 211;Nomi,Takeo等人,Clinical significance and therapeuticpotential of the programmed death-1ligand / programmed death-1 pathway in humanpancreatic cancer.ClinicalCancer Research(2007);13:2151 - 2157;Ohigashi,Yuichiro等人,Clinical significance of programmed death-1ligand-1 andprogrammed death-1ligand 2 expression in human esophageal cancer.Clin.CancerResearch(2005):11:2947 - 2953;Inman,Brant A等人,PD-L1(B7-H1)expression byurothelial carcinoma of the bladder and BCG-induced granulomata:associationswith localized stage progression.Cancer(2007):109:1499 - 1505;Shimauchi,Takatoshi等人,Augmentedexpression of programmed death-1 in both neoplasmaticand nonneoplastic CD4+ T-cells in adult T-cell Leukemia / Lymphoma.Int.J.Cancer(2007):121:2585 - 2590;Gao,Qiang等人,Overexpression of PD-L1 significantlyassociates with tumor aggressiveness and postoperative recurrence in humanhepatocellular carcinoma.Clinical Cancer Research (2009) 15: 971-979; Nakanishi, Juro et al., Overexpression of B7-H1 (PD-L1) significantly associates with tumorgrade and postoperative prognosis in human urothelial cancers. Cancer ImmunolImmunother. (2007) 56: 1173-1182; Hino et al., Tumor cell expression of programmed cell death-1 is a prognostic factor for malignant melanoma. Cancer (2010): 00: 1-9). Similarly, PD-1 expression on tumor-infiltrating lymphocytes was found to mark dysfunctional T cells in breast cancer and melanoma (Ghebeh, Hazem et al., Foxp3+ tregs and B7-H1+ / PD-1+ T lymphocytes co-infiltrate the tumor tissues of high-risk breast cancer patients: implication for immunotherapy. BMC Cancer. 2008 Feb 23; 8: 57; Ahmadzadeh, Mojgan et al., Tumor antigen-specific CD8 Tcells infiltrating the tumor express high levels of PD-1 and are functionally impaired. Blood (2009) 114: 1537-1544) and was associated with poor prognosis in renal carcinoma (Thompson, R Houston et al., PD-1 is expressed by tumor infiltrating cells and is associated with poor outcome for patients with renal carcinoma. Clinical Cancer Research (2007) 15: 1757-1761). Therefore, it has been proposed that tumor cells expressing PD-L1 interact with T cells expressing PD-1, attenuating T cell activation and evading immune surveillance, thereby leading to impaired immune responses against the tumor.

[0015] Several monoclonal antibodies that inhibit the interaction between PD-1 and one or both of its ligands, PD-L1 and PD-L2, have been approved for the treatment of cancer. Merck & Co., Inc., Rahway, NJ, USA) is a potent humanized immunoglobulin G4 (IgG4) mAb that binds to the programmed cell death 1 (PD-1) receptor with high specificity, thereby inhibiting its interaction with programmed cell death ligand 1 (PD-L1) and programmed cell death ligand 2 (PD-L2). Based on preclinical in vitro data, pembrolizumab has high affinity and potent receptor blocking activity for PD-1. (pembrolizumab) is indicated for the treatment of patients across multiple indications and is indicated for the first-line treatment of patients with unresectable or metastatic CRC (microsatellite instability-high or mismatch repair-deficient tMSI-H / dMMR). Pembrolizumab is the current standard of care for first-line MSI-H / dMMR mCRC.

[0016] Urothelial carcinoma

[0017] According to the International Agency for Research on Cancer (IARC), more than 165,000 patients die from urothelial cancer each year, and overall, urothelial cancer is the ninth most common cancer worldwide. Approximately 151,000 new cases of urothelial cancer are diagnosed annually in Europe, with 52,000 deaths. In Japan, over 22,000 new cases are diagnosed annually, with 7,600 deaths (Cancer Fact Sheets: All cancers excluding Non-Melanoma Skin. International Agency for Research on Cancer 2017. Retrieved from gco.iarc.fr / today / fact-sheets-cancers?cancer=29&type=0&sex=0, accessed December 19, 2017). The National Cancer Institute estimates that in the United States, over 79,000 new cases of bladder cancer were diagnosed in 2017, and over 16,800 people died from the disease (National Cancer Institute 2018).

[0018] The 5-year mortality rate of metastatic urothelial carcinoma is approximately 85% (American Cancer Society (ACS) 2016).

[0019] Urothelial carcinoma is the most common type of bladder cancer (90% of cases) and is also found in the renal pelvis (the location in the kidney where urine collects), the ureters (the tubes connecting the kidneys to the bladder), and the urothelial cells lining the urethra.

[0020] First-line therapy for metastatic urothelial carcinoma in patients with adequate renal function consists of cis-diamminedichloroplatinum (II) (cisplatin)-based combinations such as methotrexate, vinblastine, doxorubicin, and cisplatin (MVAC), or gemcitabine plus cisplatin, which exhibit objective response rates (ORRs) as high as 55%, including approximately 12% complete responses (CRs) (von der Maase 2000). Despite initial chemosensitivity, patients are not cured, and metastatic urothelial carcinoma has poor outcomes following these regimens: median time to progression is 7 months and median overall survival (OS) is 14 months. Long-term survival is poor (approximately 15%), and the prognosis is particularly grim in patients with visceral metastases, who have a five-year survival rate of 7% (von der Maase 2005; Bellmunt 2011).

[0021] Almost half of patients with urothelial carcinoma are not suitable for cisplatin-containing chemotherapy due to impaired renal function, poor performance status, or comorbid conditions (Dash et al., Cancer (2006); 107(3): 506-13). In these patients, standard first-line treatment has not yet been established, but current options generally include cis-diamino(cyclobutane 1,1 dicarboxylate) platinum (carboplatin)-based regimens or single-agent taxanes or gemcitabine (Cathomas 2015). In this setting, long-term survival is even lower (De Santis et al., J Clin Oncol (200%; 27(33): 5634-9). In April 2017, the U.S. Food and Drug Administration (FDA) approved the anti-programmed death ligand 1 (PD-L1) immune checkpoint inhibitor (CPI) atezolizumab. It is a first-line treatment for patients with locally advanced or metastatic urothelial carcinoma (Ia / mUC) who are ineligible for cisplatin. The fast-track approval was based on an open-label, single-arm study that showed a long duration of response, suggesting activity in a refractory population with an objective response rate (ORR) of 23%, which was similar across target expression levels (Balar 2017). The median OS in these patients was 15.9 months, although this study was a single-arm study and any OS benefit needs to be confirmed in a randomized experience (Balar et al., Lancet (2017); 389(10064): 67-76).

[0022] In May 2017, pembrolizumab It received expedited FDA approval as a first-line treatment for patients with la / mUC who are ineligible for cisplatin. The study underlying the approval demonstrated an ORR of 29%, with the median duration of response not reached at the time of analysis (median follow-up of 7.8 months) (Pembrolizumab Prescribing Information, Merck Sharp and Dohme Corp., 2017). In May 2018, the FDA issued a warning that patients with low PD-L1 expression were associated with decreased survival when treated with pembrolizumab or atezolizumab in the first-line setting compared with platinum-based chemotherapy. Subsequently, the prescribing information for these two CPIs was revised to require high PD-L1 expression in patients with metastatic urothelial carcinoma who are eligible for first-line platinum-based chemotherapy. This development further limited options for patients with metastatic urothelial carcinoma who have low PD-L1 expression.

[0023] Other options for patients who are not candidates for first-line cisplatin typically include carboplatin-based regimens or single-agent taxanes or gemcitabine (Cathomas et al., Hematol Oncol Clin North Am (2015); 29(2): 329-40).

[0024] There are very few options available for second-line treatment of metastatic disease. In the EU, the small molecule tubulin inhibitor vinflunine It was licensed in 2009 based on its modest activity (overall response rate 9%), moderate benefit in 2-month survival (6.9 months with vinflunine plus best supportive care (BSC) versus 4.6 months with BSC alone, hazard ratio 0.88), and favorable safety profile (Bellmunt et al., Clin Oncol (200%); 27(27): 4454-61). In May 2016, the FDA granted fast-track approval to atezolizumab in the United States as first salvage therapy for la / mUC following platinum therapy, and this therapy was subsequently approved in the European Union in September 2017. In February 2017, nivolumab was approved for nivolumab in the United States. It became the second immunotherapy to receive FDA fast-track approval, and was subsequently approved by the European Union in June 2017. In March and May 2017, the FDA approved avelumab, and durvalumab (Imfinzi TM ) received rapid approval, and these two PD-L1 blocking antibodies are indicated for the treatment of patients with locally advanced or metastatic urothelial carcinoma whose disease has progressed during or after platinum-containing chemotherapy, or whose disease has progressed within 12 months of neoadjuvant or adjuvant treatment with platinum-containing chemotherapy. In May 2017, pembrolizumab received regular FDA approval as a second-line treatment (Keytruda Prescribing Information, Merck, May 2017). The approval was based on the first randomized experience reported for CPI in the setting of locally advanced or metastatic post-platinum urothelial carcinoma, a phase 3 study of 542 patients that showed an OS of 10.3 months, compared to 7.4 months for taxane chemotherapy or vinflunine. In addition, the ORR for pembrolizumab was 21%, while the ORR for chemotherapy was 11%. No statistically significant difference in progression-free survival (PFS) was observed between the two groups (Bellmunt et al., N Engl J Med (2017); 376(11): 1015-26). It received EU approval in September 2017 and Japanese approval in January 2018 for the same indication. Other programmed cell death protein 1 (PD-1) and PD-L1 inhibitors are currently being evaluated in clinical trials for urothelial carcinoma as first-line and second-line therapies (Mullane et al., Curr Opin Urol (2016); 26(6): 556-63).

[0025] Although CPI provides a new approach for treating metastatic urothelial carcinoma, a small number of patients have had tumor responses and long-term survival has only been extended by a few months. For example, in May 2017, Roche announced that the validation phase 3 trial of second-line atezolizumab failed to reach its primary endpoint of OS (Roche, press release "Roche provides update on phase III study of Tecentriq (atezolizumab) in people with previously treated advanced bladder cancer", May 10, 2017). Most patients with locally advanced or metastatic urothelial carcinoma do not respond to CPIs and many responsive patients eventually experience disease progression (Rosenberg et al., Lancet (2016); 387 (10031): 1909-20). Novel treatments are still needed, particularly for patients who have not responded to CPIs or have progressed after CPI therapy.

[0026] The lack of approved first-line therapies for patients with metastatic urothelial carcinoma and the limited activity observed with second-line chemotherapy highlight the significant unmet medical need in this population.

[0027] bladder cancer

[0028] Bladder cancer accounts for approximately 5% of all new cancer cases in the United States (the fifth most common neoplasm) in men and 3% (the eighth most common neoplasm) in women. The incidence rate is slowly increasing with the aging population. The American Cancer Society (cancer.org) estimates that there are 81,400 new cases each year, including 62,100 in men and 19,300 in women, representing 4.5% of all cancer cases. The age-adjusted incidence rate in the United States is 20 per 100,000 people for both men and women. An estimated 17,980 people die from bladder cancer each year (13,050 in men and 4,930 in women), accounting for 3% of cancer-related deaths. Bladder cancer incidence and mortality rates increase significantly with age and will become an increasingly serious problem as the population becomes older. Globally, approximately 580,000 people will be diagnosed with bladder cancer in 2020, and bladder cancer will cause approximately 210,000 deaths worldwide.

[0029] Most bladder cancers recur in the bladder. Bladder cancer is managed via a combination of transurethral resection (TUR) of the bladder and intravesical chemotherapy or immunotherapy. The multifocal and recurrent nature of bladder cancer points to the limitations of TUR. TUR alone cannot cure most muscle-invasive cancers. Radical cystectomy and urethral diversion are the most effective ways to eliminate the cancer, but have an undeniable impact on urinary and sexual function. There continues to be a significant need for treatment modalities that benefit bladder cancer patients.

[0030] There is a significant need for additional treatments for urothelial and bladder cancers. These approaches include the use of antibodies and antibody drug conjugates, including in combination with other agents, as therapeutic modalities. 3. Summary of the Invention

[0031] Provided herein are methods of treating various cancers in human subjects, including methods of treating patients with unresectable locally advanced or metastatic urothelial carcinoma who are unable to receive cisplatin-based chemotherapy with an antibody drug conjugate (ADC) that binds to the 191P4D12 protein in combination with pembrolizumab.

[0032] In certain embodiments, the human subject treated with the methods provided herein is not suitable for cisplatin treatment and has not previously been treated with an immune checkpoint inhibitor (CPI), e.g., a PD-1 inhibitor, a PD-L1 inhibitor, or a PD-L2 inhibitor (including but not limited to atezolizumab, pembrolizumab, nivolumab, durvalumab, or avelumab).

[0033] Embodiment 1. A method of treating cancer in a human subject, the method comprising administering to the subject:

[0034] (a) an effective amount of an antibody-drug conjugate (ADC) comprising an anti-191P4D12 antibody or an antigen-binding fragment thereof and

[0035] (b) an effective amount of an anti-PD-1 antibody;

[0036] wherein the anti-191P4D12 antibody or antigen-binding fragment thereof binds to 191P4D12 and is coupled to one or more units of monomethyl auristatin E (MMAE);

[0037] wherein the anti-PD-1 antibody comprises: (i) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 of SEQ ID NOs: 24, 25, and 26, respectively, and (ii) a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 of SEQ ID NOs: 29, 30, and 31, respectively;

[0038] wherein the anti-191P4D12 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;

[0039] wherein the subject has urothelial carcinoma or bladder cancer;

[0040] wherein the subject has not received immune checkpoint inhibitor (CPI) therapy; and

[0041] wherein the subject is not suitable for treatment with cisplatin (cisplatin ineligible).

[0042] Embodiment 2. The method of embodiment 1, wherein the subject has visceral metastases.

[0043] Embodiment 3. The method of embodiment 1, wherein the subject has lymph node-only disease.

[0044] Embodiment 4. The method of any one of embodiments 1 to 3, wherein the site of origin of the disease is in the upper urinary tract.

[0045] Embodiment 5. The method of any one of embodiments 1 to 3, wherein the site of origin of the disease is in the lower urinary tract.

[0046] Embodiment 6. The method of any one of embodiments 1 to 5, wherein the subject has a combined positive score (CPS) for PD-L1 expression greater than or equal to 10.

[0047] Embodiment 7. The method of any one of Embodiments 1 to 5, wherein the subject has a PD-L1 expression CPS of less than 10.

[0048] Embodiment 8. The method of any one of embodiments 1 to 6, wherein the subject has a Connexin-4H-Fraction between 0 and 300.

[0049] Embodiment 9. The method of any one of embodiments 1 to 7, wherein the subject has a Connexin-4H-Fraction between 0 and 200.

[0050] Embodiment 10. The method of any one of embodiments 1 to 9, wherein the subject has an ECOG performance status of 1 to 2.

[0051] Embodiment 11. The method of any one of embodiments 1 to 9, wherein the subject has one or more conditions selected from the group consisting of: an ECOG performance status of 2, impaired renal function, and hearing loss of grade 2 or higher.

[0052] Embodiment 12. The method of any one of embodiments 1 to 9, wherein the subject has NYHA class III heart failure.

[0053] Embodiment 13. The method of embodiment 11, wherein the subject has an ECOG performance status of 2, and

[0054] The subject has (i) hemoglobin ≥ 10 g / dL; (ii) GFR ≥ 50 mL / min; and (iii) does not have NYHA class III heart failure.

[0055] Embodiment 14. The method of embodiment 11, wherein the impaired renal function is determined based on a creatinine clearance (CrCl) of less than 60 mL / min.

[0056] Embodiment 15. The method of embodiment 11, wherein the impaired renal function is determined based on a CrCl of less than 60 but not less than 30 mL / min.

[0057] Embodiment 16. The method of embodiment 11, wherein the impaired renal function is determined based on a CrCl of less than 30 but not less than 15 mL / min.

[0058] Embodiment 17. The method of any one of embodiments 1 to 16, wherein the subject has one or more conditions selected from the group consisting of:

[0059] (i) absolute neutrophil count not less than 1500 / μL;

[0060] (ii) platelet count not less than 100,000 / μL;

[0061] (iii) hemoglobin level not less than 9 g / dL;

[0062] (iv) serum bilirubin not exceeding 1.5 times the upper limit of normal (ULN) or not exceeding 3 times the ULN for patients with Gilbert's disease;

[0063] (v) CrCl not less than 30 mL / min, and

[0064] (vi) Alanine aminotransferase and aspartate aminotransferase not exceeding 3 times the ULN.

[0065] Embodiment 18. The method of embodiment 17, wherein the subject has all of conditions (i) to (vi) of embodiment 15.

[0066] Embodiment 19. The method of any one of embodiments 14 to 18, wherein the CrCl is measured by 24-hour urine collection or estimated according to Cockcroft-Gault criteria.

[0067] Embodiment 20. The method of any one of embodiments 1 to 19, wherein the subject does not suffer from sensory or motor neuropathy greater than grade 2.

[0068] Embodiment 21. The method of any one of embodiments 1 to 20, wherein the subject does not have active central nervous system metastases.

[0069] Embodiment 22. The method of any one of embodiments 1 to 21, wherein the subject does not have uncontrolled diabetes.

[0070] Embodiment 23. The method of embodiment 22, wherein the uncontrolled diabetes is determined by a hemoglobin A1c (HbA1c) of not less than 8% or a HbA1c between 7% and 8% accompanied by associated diabetic symptoms not otherwise specified.

[0071] Embodiment 24. The method of embodiment 23, wherein the relevant diabetic symptoms comprise or consist of polyuria, polydipsia, or both polyuria and polydipsia.

[0072] Embodiment 25. The method of any one of embodiments 1 to 24, wherein the subject has locally advanced or metastatic urothelial carcinoma.

[0073] Embodiment 26. The method of any one of embodiments 1 to 25, wherein the subject has locally advanced or metastatic bladder cancer.

[0074] Embodiment 27. The method of any one of embodiments 1 to 26, wherein the anti-191P4D12 antibody or antigen-binding fragment thereof comprises: a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 9, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 10, a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 11; a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 12, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 13, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 14, or

[0075] The anti-191P4D12 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, and 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.

[0076] Embodiment 28. The method of any one of embodiments 1 to 26, wherein the anti-191P4D12 antibody or antigen-binding fragment thereof comprises: a CDR-H1 consisting of the amino acid sequence of SEQ ID NO: 9, a CDR-H2 consisting of the amino acid sequence of SEQ ID NO: 10, a CDR-H3 consisting of the amino acid sequence of SEQ ID NO: 11; a CDR-L1 consisting of the amino acid sequence of SEQ ID NO: 12, a CDR-L2 consisting of the amino acid sequence of SEQ ID NO: 13, and a CDR-L3 consisting of the amino acid sequence of SEQ ID NO: 14, or

[0077] The anti-191P4D12 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, and 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.

[0078] Embodiment 29. The method of any one of embodiments 1 to 28, wherein the anti-191P4D12 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.

[0079] Embodiment 30. The method of any one of embodiments 1 to 29, wherein the anti-191P4D12 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.

[0080] Embodiment 31. The method of any one of Embodiments 1 to 30, wherein the anti-191P4D12 antigen-binding fragment is Fab, F(abt)2, Fv, or scFv.

[0081] Embodiment 32. The method of any one of embodiments 1 to 30, wherein the anti-191P4D12 antibody is a fully human antibody.

[0082] Embodiment 33. The method of any one of embodiments 1 to 30 and 32, wherein the anti-191P4D12 antibody is IgG1 and the light chain is a kappa light chain.

[0083] Embodiment 34. The method of any one of embodiments 1 to 33, wherein the anti-191P4D12 antibody or antigen-binding fragment thereof is recombinantly produced.

[0084] Embodiment 35. The method of any one of embodiments 1 to 34, wherein the anti-191P4D12 antibody or antigen-binding fragment is coupled to each unit of MMAE via a linker.

[0085] Embodiment 36. The method of embodiment 35, wherein the linker is an enzymatically cleavable linker, and wherein the linker forms a bond with a sulfur atom of the antibody or antigen-binding fragment thereof.

[0086] Embodiment 37. The method of embodiment 35 or 36, wherein the linker has the formula: -Aa-Ww-Yy-; wherein -A- is a Stretcher unit, a is 0 or 1; -W- is an Amino Acid unit, w is an integer in the range of 0 to 12; and -Y- is a Spacer unit, y is 0, 1 or 2.

[0087] Embodiment 38. The method of embodiment 37, wherein the Extender unit has the structure of the following formula (1); the Amino Acid unit is Valine-Citrulline; and the Spacer unit is a PAB group comprising the structure of the following formula (2):

[0088]

[0089] Embodiment 39. The method of embodiment 37 or 38, wherein the Stretcher unit forms a bond with a sulfur atom of the antibody or antigen-binding fragment thereof; and wherein the Spacer unit is linked to MMAE via a carbamate group.

[0090] Embodiment 40. The method of any one of embodiments 1 to 39, wherein the ADC comprises 1 to 20 MMAE units per antibody or antigen-binding fragment thereof.

[0091] Embodiment 41. The method of any one of Embodiments 1 to 40, wherein the ADC comprises 1 to 10 MMAE units per antibody or antigen-binding fragment thereof.

[0092] Embodiment 42. The method of any one of embodiments 1 to 41, wherein the ADC comprises 2 to 8 MMAE units per antibody or antigen-binding fragment thereof.

[0093] Embodiment 43. The method of any one of Embodiments 1 to 42, wherein the ADC comprises 3 to 5 MMAE units per antibody or antigen-binding fragment thereof.

[0094] Embodiment 44. The method of any one of Embodiments 1 to 43, wherein the ADC has the structure:

[0095]

[0096] wherein L- represents the anti-191P4D12 antibody or an antigen-binding fragment thereof and p is 1 to 10.

[0097] Embodiment 45. The method of Embodiment 44, wherein p is 2 to 8.

[0098] Embodiment 46. The method of Embodiment 44 or 45, wherein p is 3 to 5.

[0099] Embodiment 47. The method of any one of Embodiments 44 to 46, wherein p is 3 to 4.

[0100] Embodiment 48. The method of any one of Embodiments 44 to 47, wherein p is about 4.

[0101] Embodiment 49. The method of any one of Embodiments 44 to 47, wherein the effective amount of the antibody drug conjugate has a mean p-value of about 3.8.

[0102] Embodiment 50. The method of any one of Embodiments 1 to 49, wherein the ADC is administered to the subject at a dose of 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.

[0103] Embodiment 51. The method of any one of Embodiments 1 to 50, wherein the ADC is administered to the subject at a dose of 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.

[0104] Embodiment 52. The method of any one of embodiments 1 to 51, wherein the ADC is administered to the subject at a dose of about 1 mg / kg of the subject's body weight.

[0105] Embodiment 53. The method of any one of Embodiments 1 to 51, wherein the ADC is administered to the subject at a dose of about 1.25 mg / kg of the subject's body weight.

[0106] Embodiment 54. The method of any one of embodiments 1 to 53, wherein the ADC is administered to the subject by intravenous (IV) injection or infusion.

[0107] Embodiment 55. The method of any one of embodiments 1 to 54, wherein the ADC is administered to the subject by IV injection or infusion on up to 2 days within a 21-day treatment cycle.

[0108] Embodiment 56. The method of any one of Embodiments 1 to 55, wherein the ADC is administered to the subject by IV injection or infusion on Days 1 and 8 of a 21-day treatment cycle.

[0109] Embodiment 57. The method of any one of Embodiments 1 to 56, wherein the ADC is administered to the subject by IV injection or infusion over about 30 minutes on up to 2 days within a 21-day treatment cycle.

[0110] Embodiment 58. The method of any one of embodiments 1 to 57, wherein the ADC is administered by IV injection or infusion over about 30 minutes on days 1 and 8 of a 21-day treatment cycle.

[0111] Embodiment 59. The method of any one of embodiments 1 to 58, wherein the ADC is formulated as a pharmaceutical composition comprising L-histidine, polysorbate-20 (TWEEN-20), and trehalose dihydrate.

[0112] Embodiment 60. The method of any one of Embodiments 1 to 59, wherein the ADC is formulated as 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 wherein the pharmaceutical composition has a pH of about 6.0 at 25°C.

[0113] Embodiment 61. The method of any one of Embodiments 1 to 59, wherein the ADC is formulated as 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 pharmaceutical composition has a pH of about 6.0 at 25°C.

[0114] Embodiment 62. The method of any one of Embodiments 1 to 61, wherein the ADC has the structure:

[0115]

[0116] wherein L- represents the antibody or antigen-binding fragment thereof and p is from about 3 to about 4, the anti-191P4D12 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, wherein the ADC is administered at a dose of about 1.25 mg / kg of the subject's body weight, and wherein the dose is administered by IV injection or infusion over about 30 minutes on days 1 and 8 of a 21-day treatment cycle.

[0117] Embodiment 63. The method of any one of Embodiments 1 to 60, wherein the anti-PD-1 antibody is administered to the subject at a dose of about 100 mg to about 400 mg.

[0118] Embodiment 64. The method of any one of Embodiments 1 to 61, wherein the anti-PD-1 antibody is administered to the subject at a dose of about 200 mg.

[0119] Embodiment 65. The method of Embodiment 62, wherein:

[0120] (a) administering the anti-PD-1 antibody to the subject at a dose of about 200 mg; and

[0121] (b) After step (a), the anti-PD-1 antibody is administered to the subject at a dose of about 400 mg every 42 days.

[0122] Embodiment 66. The method of any one of Embodiments 1 to 65, wherein the anti-PD-1 antibody is administered to the subject by IV infusion on day 1 of a 21-day treatment cycle.

[0123] Embodiment 67. The method of any one of Embodiments 1 to 66, wherein the anti-PD-1 antibody is administered to the subject by IV infusion on day 1 of each 21-day treatment cycle.

[0124] Embodiment 68. The method of any one of Embodiments 1 to 67, wherein the anti-PD-1 antibody is administered to the subject by IV infusion over about 30 minutes on day 1 of a 21-day treatment cycle.

[0125] Embodiment 69. The method of any one of Embodiments 1 to 66, wherein the anti-PD-1 antibody is administered to the subject by IV infusion over about 30 minutes on Day 1 of each 21-day treatment cycle.

[0126] Embodiment 70. The method of any one of Embodiments 1 to 69, wherein the subject has a complete response following treatment.

[0127] Embodiment 71. The method of any one of Embodiments 1 to 69, wherein the subject has a partial response following treatment.

[0128] Embodiment 72. The method of any one of Embodiments 1 to 69, wherein the subject has a complete response or a partial response following treatment.

[0129] Embodiment 73. The method of any one of Embodiments 1 to 69, wherein the subject's disease is stable following treatment.

[0130] Embodiment 74. The method of any one of Embodiments 1 to 69, wherein the subject has a duration of response of at least or about 6 months after treatment.

[0131] Embodiment 75. The method of any one of Embodiments 1 to 69, wherein the subject has a duration of response of at least or about 12 months after treatment.

[0132] Embodiment 76. The method of any one of Embodiments 1 to 69, wherein the subject has a duration of response of at least or about 24 months after treatment.

[0133] Embodiment 77. The method of any one of Embodiments 1 to 69, wherein the subject has a duration of response of at least or about 27 months after treatment.

[0134] Embodiment 78. The method of any one of Embodiments 1 to 69, wherein the subject has a progression-free survival of at least or about 6 months after treatment.

[0135] Embodiment 79. The method of any one of Embodiments 1 to 69, wherein the subject has a progression-free survival of at least or about 8 months after treatment.

[0136] Embodiment 80. The method of any one of Embodiments 1 to 69, wherein the subject has a progression-free survival of at least or about 12 months after treatment.

[0137] Embodiment 81. The method of any one of Embodiments 1 to 69, wherein the subject has a progression-free survival of at least or about 20 months after treatment.

[0138] Embodiment 82. The method of any one of Embodiments 1 to 69, wherein the subject has a progression-free survival of at least or about 29 months after treatment.

[0139] Embodiment 83. The method of any one of Embodiments 1 to 69, wherein the subject has an overall survival of at least or about 22 months after treatment.

[0140] Embodiment 84. The method of any one of Embodiments 1 to 69, wherein the subject has an overall survival of at least or about 27 months after treatment.

[0141] Embodiment 85. The method of any one of Embodiments 1 to 69, wherein the subject has an overall survival of at least or about 30 months after treatment.

[0142] Embodiment 86. The method of any one of Embodiments 1 to 69, wherein the subject has an overall survival in the range of 19 to 25 months following treatment.

[0143] Embodiment 87. The method of any one of Embodiments 1 to 69, wherein the subject has an overall survival in the range of 28 to 32 months following treatment.

[0144] Embodiment 88. The method of any one of embodiments 1 to 69, wherein a population of subjects is treated by the method, and wherein the percentage of subjects with a complete response in the treated population is at least or about 10%.

[0145] Embodiment 89. The method of any one of embodiments 1 to 69, wherein a population of subjects is treated by said method, and wherein the percentage of subjects with a partial response in the treated population is at least or about 54%.

[0146] Embodiment 90. The method of any one of embodiments 1 to 69, wherein a population of subjects is treated by said method, and wherein the objective response rate in the treated population is at least or about 65%.

[0147] Embodiment 91. The method of any one of embodiments 1 to 69, wherein a population of subjects is treated by said method, and wherein the objective response rate in the treated population is in the range of 53% to 75%.

[0148] Embodiment 92. The method of any one of embodiments 1 to 69, wherein a population of subjects is treated by said method, and wherein the percentage of subjects with stable disease in the treated population is at least or about 22%.

[0149] Embodiment 93. The method of any one of embodiments 1 to 69, wherein a population of subjects is treated by said method, and wherein the duration of response in the treated population is at least or about 6 months.

[0150] Embodiment 94. The method of any one of embodiments 1 to 69, wherein a population of subjects is treated by said method, and wherein the duration of response in the treated population is at least or about 12 months.

[0151] Embodiment 95. The method of any one of embodiments 1 to 69, wherein a population of subjects is treated by said method, and wherein the duration of response in the treated population is at least or about 24 months.

[0152] Embodiment 96. The method of any one of embodiments 1 to 69, wherein a population of subjects is treated by said method, and wherein the duration of response in the treated population is at least or about 27 months.

[0153] Embodiment 97. The method of any one of Embodiments 1 to 69, wherein a population of subjects is treated by said method, and wherein the progression-free survival in the treated population is at least or about 6 months.

[0154] Embodiment 98. The method of any one of embodiments 1 to 69, wherein a population of subjects is treated by said method, and wherein the progression-free survival in the treated population is at least or about 12 months.

[0155] Embodiment 99. The method of any one of embodiments 1 to 69, wherein a population of subjects is treated by said method, and wherein the progression-free survival in the treated population is at least or about 20 months.

[0156] Embodiment 100. The method of any one of embodiments 1 to 69, wherein a population of subjects is treated by said method, and wherein the progression-free survival in the treated population is at least or about 29 months.

[0157] Embodiment 101. The method of any one of Embodiments 1 to 69, wherein a population of subjects is treated by said method, and wherein the median overall survival in the treated population is at least or about 22 months.

[0158] Embodiment 102. The method of any one of embodiments 1 to 69, wherein a population of subjects is treated by the method, and wherein the median overall survival in the treated population is at least or about 27 months.

[0159] Embodiment 103. The method of any one of embodiments 1 to 69, wherein a population of subjects is treated by the method, and wherein the median overall survival in the treated population is at least or about 30 months.

[0160] Embodiment 104. The method of any one of embodiments 1 to 69, wherein a population of subjects is treated by said method, and wherein the overall survival in the treated population is in the range of 19 to 25 months.

[0161] Embodiment 105. The method of any one of embodiments 1 to 69, wherein a population of subjects is treated by said method, and wherein the overall survival in the treated population is in the range of 30 to 32 months.

[0162] Embodiment 106. The method of any one of embodiments 1 to 70 and 72, wherein the complete response rate in a population of subjects treated with the method is at least or about 10%.

[0163] Embodiment 107. The method of any one of embodiments 1 to 69, 71 and 72, wherein the partial response rate in the population of subjects treated with the method is at least or about 54%.

[0164] Embodiment 108. The method of any one of embodiments 1 to 72, wherein the objective response rate in a population of subjects treated with the method is at least or about 65%.

[0165] Embodiment 109. The method of any one of embodiments 1 to 72, wherein the objective response rate in the population of subjects treated with said method is 53% to 75%.

[0166] Embodiment 110. The method of any one of embodiments 1 to 69 and 73, wherein the rate of disease stabilization in a population of subjects treated with the method is at least or about 22%.

[0167] Embodiment 111. The method of any one of Embodiments 1 to 69 and 74 to 77, wherein the duration of response in the subject population treated with the method is at least or about 6 months.

[0168] Embodiment 112. The method of any one of Embodiments 1 to 69 and 74 to 77, wherein the duration of response in the subject population treated with the method is at least or about 12 months.

[0169] Embodiment 113. The method of any one of Embodiments 1 to 69 and 74 to 77, wherein the duration of response in the subject population treated with the method is at least or about 24 months.

[0170] Embodiment 114. The method of any one of Embodiments 1 to 69 and 74 to 77, wherein the duration of response in the population of subjects treated with the method is at least or about 27 months.

[0171] Embodiment 115. The method of any one of Embodiments 1 to 69 and 78 to 82, wherein the population of subjects treated with the method has a progression-free survival of at least or about 6 months.

[0172] Embodiment 116. The method of any one of Embodiments 1 to 69 and 78 to 82, wherein the progression-free survival of the subject population treated with the method is at least or about 12 months.

[0173] Embodiment 117. The method of any one of Embodiments 1 to 69 and 78 to 82, wherein the progression-free survival of the subject population treated with the method is at least or about 20 months.

[0174] Embodiment 118. The method of any one of Embodiments 1 to 69 and 78 to 82, wherein the progression-free survival of the population of subjects treated with the method is at least or about 29 months.

[0175] Embodiment 119. The method of any one of Embodiments 1 to 69 and 83 to 87, wherein the median overall survival of the population of subjects treated with the method is at least or about 22 months.

[0176] Embodiment 120. The method of any one of embodiments 1 to 69 and 83 to 87, wherein the median overall survival of the population of subjects treated with the method is at least or about 27 months.

[0177] Embodiment 121. The method of any one of Embodiments 1 to 69 and 83 to 87, wherein the median overall survival of the population of subjects treated with the method is at least or about 30 months.

[0178] Embodiment 122. The method of any one of embodiments 1 to 69, 79 and 80, wherein the overall survival of the subject population treated with said method is 19 to 25 months.

[0179] Embodiment 123. The method of any one of embodiments 1 to 69, 79 and 80, wherein the overall survival of the subject population treated with said method is 30 to 32 months. 4. Description of the Figures

[0180] Figures 1A-1E The nucleotide and amino acid sequences of the connexin-4 protein are depicted ( Figure 1A ), the heavy chain of Ha22-2(2.4)6.1 ( Figure 1B ) and light chain ( Figure 1C ) nucleotide and amino acid sequences, and the heavy chain of Ha22-2(2.4)6.1 ( Figure 1D ) and light chain ( Figure 1E )'s amino acid sequence.

[0181] Figure 2 Describes the overall study design for the clinical studies described in Section 6.1.

[0182] Figure 3 Describes the investigational phase of a Phase 1b / 2 study of enfortumab vedotin in combination with pembrolizumab as first-line treatment in patients with unresectable locally advanced or metastatic urothelial carcinoma (1a / mUC) who are unable to receive cisplatin-based chemotherapy, as described in Section 6.1.

[0183] Figure 4 The European Organisation for Research and Treatment of Cancer (EORTC) Core Quality of Life (QLQ-C-30) assessment (EORTC-QLQ-C-30, current version, version 3) was depicted as described in Section 6.1.

[0184] Figure 5 The EuroQol-5 dimensions (EQ-5D-5L) described in Section 6.1 are depicted.

[0185] Figure 6 Tumor reduction (ie, tumor size (% change from baseline)) is depicted for individual patients in the clinical studies, as described in Section 6.1.

[0186] Figure 7 Depicts the percent change from baseline in the sum of target lesion diameters over time (in months) based on blinded independent central review in clinical studies, as described in Section 6.1.

[0187] Figure 8Depicted are subgroup analyses of ORR for patients administered the combination of enrofloxacin and pembrolizumab (ie, EV+pembro arm) in the clinical study described in Section 6.1.

[0188] Figure 9 Depicted is a subgroup analysis of ORR in patients administered enrotumomab vedotin monotherapy (ie, the EVMono arm) in the clinical study described in Section 6.1.

[0189] Figure 10 Depicted are H-scores and best responses based on blinded independent central review of connexin-4 expression at baseline in clinical studies, as described in Section 6.1.

[0190] Figure 11 Depicted are H-scores and best overall responses based on blinded independent central review of connexin-4 expression at baseline in clinical studies, as described in Section 6.1.

[0191] Figure 12 Duration of response (DOR) based on blinded independent central review in the clinical studies described in Section 6.1 is depicted.

[0192] Figure 13 Depicts progression-free survival rates based on blinded independent central review in the clinical studies described in Section 6.1.

[0193] Figure 14 Depict overall survival rates from the clinical studies described in Section 6.1.

[0194] Figure 15 Duration of response (DOR) according to blinded independent central review in the clinical studies described in Section 6.2 is depicted.

[0195] Figure 16 Depicts progression-free survival rates based on blinded independent central review in the clinical studies described in Section 6.2.

[0196] Figure 17 Depict overall survival rates from the clinical studies described in Section 6.2. 5. Specific implementation methods

[0197] Before the present disclosure is further described, it is to be understood that this disclosure is not limited to the particular embodiments set forth herein, and it is to be further understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.

[0198] 5.1 Definition

[0199] The techniques and procedures described or referenced herein include those that are substantially well understood and / or commonly used by those skilled in the art using routine methods, such as the widely used methods described in the following references: 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, Volumes 1 and 2 (Kontermann and Dübel eds., 2nd ed., 2010).

[0200] Unless otherwise defined herein, the technical and scientific terms used in this specification shall have the meanings commonly understood by those of ordinary skill in the art. For the purpose of interpreting this specification, the following terminology will apply and, whenever appropriate, terms used in the singular will also include the plural and vice versa. In the event of a conflict between any description of a term set forth and any document incorporated herein by reference, the terminology set forth below shall prevail.

[0201] The terms "antibody," "immunoglobulin," or "Ig" are used interchangeably herein and are used in the broadest sense and specifically encompass, for example, monoclonal antibodies (including agonists, antagonists, neutralizing antibodies, full-length or intact monoclonal antibodies), antibody compositions with multiple or single epitope specificity, polyclonal or monovalent antibodies, multivalent antibodies, multispecific antibodies formed from at least two intact antibodies (e.g., bispecific antibodies, as long as they exhibit the desired biological activity), single-chain antibodies, and fragments thereof, as described below. The antibodies can be human, humanized, chimeric, and / or affinity matured, as well as antibodies from other species (e.g., mouse, rabbit, etc.). The term "antibody" is intended to include polypeptide products of B cells within the immunoglobulin class of polypeptides that are capable of binding to specific molecular antigens and are composed of two pairs of identical polypeptide chains, each pair having a heavy chain (about 50-70 kDa) and a light chain (about 25 kDa), each amino-terminal portion of each chain including a variable region of about 100 to about 130 or more amino acids, and each carboxyl-terminal portion of each chain including a constant region. See, for example, Antibody Engineering (Borrebaeck, ed., 2nd ed., 1995); and Kuby, Immunology (3rd ed., 1997). In specific embodiments, a specific molecular antigen can be bound by the antibodies (including polypeptides or epitopes) provided herein. Antibodies also include, but are not limited to, synthetic antibodies, recombinantly produced antibodies, camelized antibodies, intracellular antibodies, anti-idiotype (anti-Id) antibodies, and functional fragments (e.g., antigen-binding fragments) of any of the above, which refers to portions of antibody heavy or light chain polypeptides that retain 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 Fv (scFv) (e.g., including monospecific, bispecific, etc.), Fab fragments, F(ab') fragments, F(ab')2 fragments, F(ab')2 fragments, disulfide-linked Fv (dsFv), Fd fragments, Fv fragments, diabodies, triabodies, tetrabodies, and minibodies. In particular, the antibodies provided herein include immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, e.g., antigen-binding domains or molecules containing an antigen-binding 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; Plückthun and Skerra, 1989, Meth. Enzymol. 178: 497-515; and Day, Advanced Immunochemistry (2nd ed. 1990). The antibodies provided herein can belong to any class (e.g., IgG, IgE, IgM, IgD, and IgA) of immunoglobulin molecules or any subclass (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2). The antibodies can be agonistic or antagonistic antibodies.

[0202] 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 naturally occurring mutations that may be present in minor amounts. Monoclonal antibodies are highly specific for a single antigenic site. In contrast to polyclonal antibody preparations, which can include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen.

[0203] An "antigen" is a structure to which an antibody can selectively bind. A target antigen can be a polypeptide, carbohydrate, nucleic acid, lipid, hapten, or other naturally occurring or synthetic compound. In some embodiments, the target antigen is a polypeptide. In certain embodiments, the antigen is associated with a cell, for example, present on or in a cell (e.g., a cancer cell).

[0204] An "intact" antibody is one that comprises an antigen binding site, 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.

[0205] The terms "antigen-binding fragment," "antigen-binding domain," "antigen-binding region," and similar terms refer to a portion of an antibody that contains the amino acid residues that interact with an antigen and impart its specificity and affinity to the binding agent for the antigen (e.g., CDRs). As used herein, "antigen-binding fragment" includes "antibody fragments," which contain a portion of an intact antibody, such as the antigen-binding region or variable region of an intact antibody. Examples of antibody fragments include, but are not limited to, Fab, Fab', F(ab')2, and Fv fragments; diabodies and di-diabodies (see, 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., 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); and multispecific antibodies formed from antibody fragments.

[0206] The term "binds" or "binding" refers to interactions between molecules, including, for example, the formation of a complex. The interaction can be, for example, a non-covalent interaction, including hydrogen bonds, ionic bonds, hydrophobic interactions, and / or van der Waals interactions. A complex can also include the binding of two or more molecules held together by covalent or non-covalent bonds, interactions, or forces. The total non-covalent interaction strength between a single antigen binding site on an antibody and a single epitope of a target molecule (e.g., an antigen) is the affinity of the antibody or functional fragment for that epitope. The dissociation rate (k) of a binding molecule (e.g., an antibody) from a monovalent antigen is the rate of dissociation of the binding molecule (e.g., an antibody) from the monovalent antigen. off ) and binding rate (k on ) ratio (k off / k on ) is the dissociation constant K D , which is negatively correlated with affinity. K D The lower the value, the higher the antibody affinity. D The value varies for different complexes of antibody and antigen and depends on the k onWith k off Both. The dissociation constant K of the antibodies provided herein D Can use any method provided herein or any other method well-known to those skilled in the art to measure.The avidity at a binding site can not always reflect the true interaction intensity between antibody and antigen.When the complex antigen (such as multivalent antigen) containing multiple repeated antigenic determinants contacts with the antibody containing multiple binding sites, the interaction of antibody and antigen at one site will increase the reaction probability at a second site.The intensity of this multiple interaction between multivalent antibody and antigen is called avidity.

[0207] In connection with the antibodies or antigen-binding fragments thereof described herein, terms such as "bind to," "that specifically bind to," and similar terms are also used interchangeably herein and refer to a binding molecule having an antigen-binding domain that specifically binds to an antigen (e.g., a polypeptide). An antibody or antigen-binding fragment that binds to or specifically binds to an antigen may have cross-reactivity with related antigens. In certain embodiments, an antibody or antigen-binding fragment that binds to or specifically binds to an antigen does not have cross-reactivity with other antigens. An antibody or antigen-binding fragment that binds to or specifically binds to an antigen may be detected, for example, by immunoassay, Or other techniques known to those skilled in the art to identify. In some embodiments, when the antibody or antigen-binding fragment binds to the antigen with a higher affinity than to any cross-reactive antigen, as determined using experimental techniques (e.g., radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA)), the antibody or antigen-binding fragment binds to or specifically binds to the antigen. Typically, a specific or selective reaction will be at least twice the background signal or noise and may exceed 10 times the background. For a discussion of binding specificity, see, for example, Fundamental Immunology 332-36 (Paul, ed., 2nd 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, for example, as determined by fluorescence activated cell sorting (FACS) analysis or RIA. For example, terms such as "specifically bind," "specifically bind to," or "specific for..." mean a binding that is measurably different from a non-specific interaction. Specific binding can be measured, for example, by measuring the binding of a molecule compared to the binding of a control molecule, which is typically a molecule of similar structure that does not have binding activity. For example, specific binding can be measured by competition with a control molecule similar to the target (e.g., an excess of unlabeled target). In this case, if the binding of the labeled target to the probe is competitively inhibited by an excess of unlabeled target, specific binding is indicated. Antibodies or antigen-binding fragments that bind to an antigen include antibodies or antigen-binding fragments that can bind to the antigen with an affinity sufficient to make the binding molecule suitable for use as, for example, a diagnostic agent in the targeted antigen. In certain embodiments, the antibody or antigen-binding fragment that binds to the antigen has a dissociation constant (K) of less than or equal to . D ): 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. In certain embodiments, the antibody or antigen-binding fragment binds to an epitope of the antigen that is conserved between antigens from different species (e.g., between humans and cynomolgus monkey species).

[0208] "Binding affinity" generally refers to the total strength 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 can generally be represented by a dissociation constant (KD). Affinity can be measured by common methods known in the art, including those described herein. Low-affinity antibodies generally bind to the antigen slowly and tend to dissociate easily, while high-affinity antibodies generally bind to the antigen faster and tend to remain bound for a longer time. A variety of methods for measuring binding affinity are known in the art, any of which can be used for the purposes of the present disclosure. Specific illustrative embodiments include the following. In one embodiment, "KD" is a measure of the affinity of a binding molecule X to its binding partner Y. D ” or “K D The "value" can be measured by assays known in the art, such as by binding assays. D It can be measured using RIA, for example, using a Fab form of the antibody of interest and its antigen (Chen et al., 1999, J. Mol Biol 293:865-81). D or K D The value can also be determined by using biolayer interferometry (BLI) or surface plasmon resonance (SPR). Use e.g. QK384 system, or through Use e.g. TM-2000 or The "on-rate" or "association rate" or "kon" can also be measured using the same biolayer interferometry (BLI) or surface plasmon resonance (SPR) techniques described above, using, for example, QK384, TM-2000 or TM-3000 system for determination.

[0209] In certain embodiments, antibodies or antigen-binding fragments may comprise "chimeric" sequences in which a portion of the heavy and / or light chain is identical or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain is identical or homologous to corresponding sequences in antibodies derived 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).

[0210] In certain embodiments, antibody or antigen-binding fragment can comprise part of the " humanized " form of non-human (such as murine) antibody, and described non-human antibody is chimeric antibody (such as acceptor antibody) comprising human immunoglobulin, wherein native CDR residues are replaced by residues of corresponding CDR from non-human species (such as mouse, rat, rabbit or non-human primate) (such as donor antibody) with desired specificity, affinity and ability. In some cases, one or more FR region residues of human immunoglobulin are replaced by corresponding non-human residues. In addition, humanized antibody can be included in residues not found in recipient antibody or donor antibody. These modifications are carried out to further optimize antibody performance. Humanized antibody heavy chain or light chain can comprise substantially all at least one or more variable regions, wherein all or substantially all CDRs correspond to the CDRs of non-human immunoglobulins, and all or substantially all FRs are FRs of human immunoglobulin sequences. In certain embodiments, humanized antibody will comprise at least a portion of immunoglobulin constant region (Fc), typically at least a portion of 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.

[0211] In certain embodiments, an antibody or antigen-binding fragment may comprise a portion of a "fully human antibody" or "human antibody," wherein the terms are used interchangeably herein and refer to antibodies comprising human variable regions and, for example, human constant regions. In a specific embodiment, the term refers to an antibody comprising a variable region and a constant region of human origin. In certain embodiments, a "fully human" antibody may also encompass an antibody that binds to a polypeptide and is encoded by a nucleic acid sequence that is a naturally occurring somatic variant of a human germline immunoglobulin nucleic acid sequence. The term "fully human antibody" includes antibodies comprising variable and constant regions corresponding to human germline immunoglobulin sequences as described by Kabat et al. (see Kabat et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No. 91-3242). A "human antibody" is an antibody having an amino acid sequence corresponding to the amino acid sequence of an antibody produced by a human and / or has been prepared using any technology for preparing human antibodies. This definition of a human antibody specifically excludes humanized antibodies comprising non-human antigen-binding residues. Human antibodies can be produced using various 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). The methods described in Cole et al., Monoclonal Antibodies and Cancer Therapy 77 (1985); Boemer et al., 1991, J. Immunol. 147 (1): 86-95; and van Dijk and van de Winkel, 2001, Curr. Opin. Pharmacol. 5: 368-74 can also be used to prepare human monoclonal antibodies. Human antibodies can be prepared by administering an antigen to a transgenic animal (e.g., a mouse) that has been modified to produce such antibodies in response to antigenic stimulation, but whose endogenous loci have been disabled (see, e.g., Jakobovits, 1995, Curr. Opin. Biotechnol. 6(5):561-66; Brüggemann and Taussing, 1997, Curr. Opin. Biotechnol. 8(4):455-58; and U.S. Pat. Nos. 6,075,181 and 6,150,584, regarding XENOMOUSE TMTechnology). See also, for example, Li et al., 2006, Proc. Natl. Acad. Sci. USA, 103: 3557-62 for human antibodies generated via human B cell hybridoma technology.

[0212] In certain embodiments, the antibody or antigen-binding fragment may comprise a portion of a "recombinant human antibody," wherein the phrase includes human antibodies prepared, expressed, generated, or isolated by recombinant means, such as antibodies expressed using a recombinant expression vector transfected into a host cell; antibodies isolated from a recombinant combinatorial human antibody library; antibodies isolated from transgenic and / or transchromosomal animals (e.g., mice or cows) 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 involving splicing of human immunoglobulin gene sequences to other DNA sequences. Such recombinant human antibodies may have variable and constant regions derived from human germline immunoglobulin sequences (see Kabat, EA et al. (1991) Sequences of Proteins of Immunological Interest, 5th ed., US Department of Health and Human Services, NIH Publication No. 91-3242). However, in certain embodiments, such recombinant human antibodies are subjected to in vitro mutagenesis (or in vivo somatic mutagenesis when animals transgenic for human Ig sequences are used) so that the amino acid sequences of the VH and VL regions of the recombinant antibodies, while derived from and related to human germline VH and VL sequences, do not naturally exist within the human antibody germline antibody repertoire in vivo.

[0213] In certain embodiments, the antibody or antigen-binding fragment may comprise a portion of a "monoclonal antibody," wherein, as used herein, the term refers to an antibody obtained from a population of substantially homogeneous antibodies, e.g., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in small amounts, and each monoclonal antibody will generally recognize a single epitope on the antigen. In specific embodiments, as used herein, a "monoclonal antibody" is an antibody produced by a single hybridoma or other cell. The term "monoclonal" is not limited to any particular method for making an antibody. For example, monoclonal antibodies suitable for use in the present disclosure can be prepared by the hybridoma method first described by Kohler et al., 1975, Nature 256:495, or can be prepared using recombinant DNA methods in bacteria or eukaryotic or plant cells (see, e.g., U.S. Patent No. 4,816,567). Monoclonal antibodies can also be isolated from phage antibody libraries using techniques such as those described in Clackson et al., 1991, Nature 352:624-28 and Marks et al., 1991, J. Mol. Biol., 222:581-97. Other methods for preparing clonal cell lines and monoclonal antibodies expressed therefrom are well known in the art. See, for example, Short Protocols in Molecular Biology (Ausubel et al., ed., 5th ed. 2002).

[0214] The typical 4-chain antibody unit is a heterotetrameric glycoprotein, which is composed of two identical light (L) chains and two identical heavy (H) chains. In the case of IgG, the 4-chain unit is generally about 150,000 daltons. Each L chain is connected to the H chain by a covalent disulfide bond, and the two H chains are connected to each other by one or more disulfide bonds depending on the H chain isotype. Each H chain and L chain also have regularly spaced intrachain disulfide bridges. Each H chain has a variable domain (VH) at the N-terminus, followed by three constant domains (CH) of each α and γ chain and four CH domains of μ and ε isotypes. Each L chain has a variable domain (VL) at the N-terminus, followed by a constant domain (CL) at its other end. VL is compared with VH, and CL is compared with the first constant domain (CH1) of the heavy chain. It is believed that specific amino acid residues form an interface between the light chain variable domain and the heavy chain variable domain. VH and VL are paired together to form a single antigen binding site. For the structure and properties of different classes of antibodies, see, eg, Basic and Clinical Immunology 71 (Stites et al., eds., 8th ed. 1994); and Immunobiology (Janeway et al., eds., 5th ed. 2001).

[0215] The term "Fab" or "Fab region" refers to the region of an antibody that binds to an antigen. Conventional IgG typically comprises two Fab regions, each residing on one of the two arms of the Y-shaped IgG structure. Each Fab region is typically composed of one variable region and one constant region of each of the heavy and light chains. More specifically, the heavy chain variable region and constant region in the Fab region are the VH and CH1 regions, and the light chain variable region and constant region in the Fab region are the VL and CL regions. The VH, CH1, VL, and CL in the Fab region can be arranged in various ways to confer antigen binding ability according to the present disclosure. For example, the VH and CH1 regions can be located on one polypeptide, and the VL and CL regions can be located on separate polypeptides, similar to the Fab regions of conventional IgG. 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.

[0216] The terms "variable region," "variable domain," "V region," or "V domain" refer to a portion of an antibody's light or heavy chain that is typically located at the amino terminus of the light or heavy chain and is about 120 to 130 amino acids in length in the heavy chain and about 100 to 110 amino acids in length in the light chain, and is responsible for the binding and specificity of each particular antibody to its specific antigen. The variable region of the heavy chain may be referred to as "VH." The variable region of the light chain may be referred to as "VL." The term "variable" refers to the fact that certain segments of the variable region differ widely in sequence among antibodies. The V region mediates antigen binding and defines the specificity of a particular antibody for its specific antigen. However, variability is not evenly distributed across the 110 amino acid span of the variable region. In practice, the V region consists of a weakly variable (e.g., relatively constant) stretch of about 15-30 amino acids, called a framework region (FR), separated by shorter, more variable (e.g., extremely variable) regions, called "hypervariable regions," each about 9-12 amino acids in length. The variable regions of the heavy and light chains each comprise four FRs that largely adopt a β-pleated structure, and the FRs are connected by three hypervariable regions, which form a loop connecting the β-pleated structure and form a part of the β-pleated structure in some cases. The hypervariable region in each chain is tightly bound together with the hypervariable regions from other chains through the FRs, promoting the formation of the antigen-binding site of the antibody (see, for example, Kabat et al., Sequences of Proteins of Immunological Interest (5th edition, 1991)). The constant region is not directly involved in the binding of the antibody to the antigen, but exhibits a variety of effector functions, such as causing the antibody to participate in antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC). The variable region is widely different in sequence between different antibodies. In a specific embodiment, the variable region is a human variable region.

[0217] The term "variable region residue numbering according to Kabat" or "amino acid position numbering as in Kabat" and variations thereof refer to the numbering system used by Kabat et al. (see above) to compile antibody heavy chain variable regions or light chain variable regions. Using this numbering system, the actual linear amino acid sequence may contain shortening of the FR or CDR corresponding to the variable domain or fewer or other amino acids inserted therein. For example, the heavy chain variable domain may include 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, etc., according to Kabat). For a given antibody, the Kabat numbering of the residues can be determined by aligning the homologous region of the antibody sequence with the "standard" Kabat numbering sequence. The Kabat numbering system is typically 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., see above). The "EU numbering system" or "EU index" (e.g., the EU index as reported in Kabat et al., supra) is often used when referring to residues in the constant region of an immunoglobulin heavy chain. The "EU index as in Kabat" refers to the residue numbering of the human IgG1 EU antibody. Other numbering systems have been described, for example, by AbM, Chothia, Contact, IMGT, and AHon.

[0218] When used in reference to antibodies, the term "heavy chain" refers to a polypeptide chain of about 50-70 kDa, wherein the amino-terminal portion includes a variable region of about 120 to 130 or more amino acids, and the carboxyl-terminal portion includes a constant region. Based on the amino acid sequence of the heavy chain constant region, the constant region can be one of five different types (e.g., isotypes), referred to as α (alpha), δ (delta), ε (epsilon), γ (gamma), and μ (mu). Different heavy chains have different sizes: α, δ, and γ contain approximately 450 amino acids, while μ and ε contain approximately 550 amino acids. When combined with a light chain, these different types of heavy chains produce the five well-known classes (e.g., isotypes) of antibodies, namely IgA, IgD, IgE, IgG, and IgM, including the four subclasses of IgG, i.e., IgG1, IgG2, IgG3, and IgG4.

[0219] When used in reference to antibodies, the term "light chain" refers to a polypeptide chain of about 25 kDa, wherein the amino-terminal portion includes a variable region of about 100 to about 110 or more amino acids, and the carboxyl-terminal portion includes a constant region. The approximate length of a light chain is 211 to 217 amino acids. Based on the amino acid sequence of the constant domain, there are two different types, designated as κ (kappa) or λ (lambda).

[0220] 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 the VH β-sheet framework of an immunoglobulin (Ig or antibody), or one of the three hypervariable regions (L1, L2, or L3) within the non-framework region of the VL β-sheet framework of an antibody. Thus, CDRs are variable region sequences interspersed within framework region sequences.

[0221] CDR regions are well known to those skilled in the art and have been defined by well-known numbering systems. For example, the Kabat complementarity determining regions (CDRs) are based on sequence variability and are most commonly used (see, e.g., Kabat et al., supra). Chothia actually refers to the position of the structural loops (see, e.g., Chothia and Lesk, 1987, J. Mol. Biol. 196:901-17). When numbering using the Kabat numbering convention, the terminus 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 the insertion at H35A and H35B; if neither 35A nor 35B is present, the loop terminus is at 32; if only 35A is present, the loop terminus is at 33; if both 35A and 35B are present, the loop terminus is at 34). The AbM hypervariable regions represent a compromise between the Kabat CDRs and the Chothia structural loops and are used by Oxford Molecular's AbM antibody modeling software (see, e.g., Antibody Engineering, Vol. 2 (Kontermann and Dübel, eds., 2nd ed., 2010)). The "contact" hypervariable regions are based on 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 specific to immunoglobulins (IGs), T cell receptors (TCRs) and major histocompatibility complexes (MHCs) of humans and other vertebrates. Herein, CDRs are referred to in terms of amino acid sequence and position within the light or heavy chain. Since the "position" of CDRs within the immunoglobulin variable domain structure is conserved between species and is present in structures called loops, CDR and framework residues are readily identified by using a numbering system that aligns variable domain sequences based on structural features. This information can be used to transplant and replace CDR residues from immunoglobulins of one species into an acceptor framework, typically from a human antibody. Another numbering system (AHon) has been developed by Honegger and Plückthun, 2001, J. Mol. Biol. 309: 657-70. The correspondence between numbering systems (including, for example, Kabat numbering and the IMGT unique numbering system) is well known to those skilled in the art (see, for example, Kabat, supra; Chothia and Lesk, supra; Martin, supra; Lefranc et al., supra). The residues from each of these hypervariable regions or CDRs are indicated in Table 1 below.

[0222] Table 1

[0223]

[0224] The boundaries of a given CDR may vary according to the scheme used to identify it. Therefore, unless otherwise specified, the terms "CDR" and "complementary determining region" or its region of a given antibody, such as a variable region, and individual CDRs (such as CDR-H1, CDR-H2) or its region of an antibody should be understood to encompass complementary determining regions as defined by any one of the known schemes described above. In some cases, it is specified to identify one or more specific CDRs, such as the CDRs defined by Kabat, Chothia or Contact methods. In other cases, the specific amino acid sequence of a CDR is provided.

[0225] The hypervariable regions may comprise the following "extended hypervariable regions": 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.

[0226] The term "constant region" or "constant domain" refers to the carboxyl-terminal portion of the light and heavy chains that is not directly involved in binding the antibody to the antigen, but exhibits various effector functions, such as interactions with Fc receptors. The term refers to the portion of the immunoglobulin molecule that contains an amino acid sequence that is more conserved relative to the other parts of the immunoglobulin (the variable region, which contains the antigen binding site). The constant region may contain the CH1, CH2, and CH3 regions of the heavy chain and the CL region of the light chain.

[0227] The term "framework" or "FR" refers to those variable region residues that flank the CDRs. FR residues are found, for example, in chimeric antibodies, humanized antibodies, human antibodies, domain antibodies, diabodies, linear antibodies, and bispecific antibodies. FR residues are those variable domain residues other than the hypervariable region residues or CDR residues.

[0228] As used herein, the term "Fc region" 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 generally defined as extending from the amino acid residue at position Cys226 or from Pro230 to its carboxyl terminus. The C-terminal lysine (residue 447, according to the EU numbering system) of the Fc region can be removed, for example, during the preparation or purification of the antibody, or by recombinantly engineering the nucleic acid encoding the heavy chain of the antibody. Thus, a composition of a complete antibody can include an antibody population in which all K447 residues have been removed, an antibody population in which the K447 residue has not been removed, and an antibody population comprising a mixture of antibodies with and without the K447 residue. A "functional Fc region" possesses the "effector functions" of the native sequence Fc region. Exemplary "effector functions" include C1q binding; CDC; Fc receptor binding; ADCC; phagocytosis; downregulation of cell surface receptors (e.g., B cell receptors), etc. Such effector functions generally require an Fc region in combination with a binding region or binding domain (e.g., an antibody variable region or domain), and can be assessed using various assays known to those skilled in the art. A "variant Fc region" comprises an amino acid sequence that differs from a native sequence Fc region by at least one amino acid modification (e.g., substitution, addition, or deletion). In certain embodiments, the variant Fc region has at least one amino acid substitution in the native sequence Fc region or in the Fc region of the parent polypeptide, e.g., about one to about ten amino acid substitutions or about one to about five amino acid substitutions, compared to the native sequence Fc region or compared to the Fc region of the parent polypeptide. The variant Fc region herein may have at least about 80% homology to the Fc region of the native sequence Fc region and / or the parent polypeptide, or at least about 90% homology thereto, e.g., at least about 95% homology thereto.

[0229] As used herein, "epitope" is a term in the art and refers to a localized region of an antigen to which a binding molecule (e.g., an antibody) can specifically bind. An epitope may be a linear epitope or may be a conformational, non-linear, or discontinuous epitope. For example, in the case of a polypeptide antigen, an epitope may be a continuous amino acid of a polypeptide (a "linear" epitope), or an epitope may comprise amino acids from two or more discontinuous regions of a polypeptide (a "conformational," "non-linear," or "discontinuous" epitope). It will be appreciated by those skilled in the art that, in general, a linear epitope may or may not be dependent on a secondary, tertiary, or quaternary structure. For example, in some embodiments, a binding molecule is bound to a group of amino acids, regardless of whether they are folded in a native three-dimensional protein structure. In other embodiments, the binding molecule requires that the amino acid residues constituting the epitope present a specific conformation (e.g., bend, twist, turn, or fold) to recognize and bind to the epitope.

[0230] The terms "polypeptide" and "peptide" and "protein" are used interchangeably herein and refer to amino acid polymers of any length. The polymer may be straight or branched, it may contain modified amino acids, and it may be interrupted by non-amino acids. The term also encompasses amino acid polymers that have been modified naturally or by intervention, such as disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation or modification. Also included within the definition are, for example, polypeptides containing one or more analogs of amino acids, including but not limited to non-natural amino acids, as well as other modifications known in the art. It should be understood that because the polypeptides of the present disclosure may be based on antibodies or other members of the immunoglobulin superfamily, in certain embodiments, a "polypeptide" may appear as a single chain or as two or more related chains.

[0231] As used herein, the term "pharmaceutically acceptable" means approved by a regulatory agency of the Federal or a state government or listed in the US Pharmacopia, the European Pharmacopia, or other generally recognized pharmacopeia for use in animals, and more particularly in humans.

[0232] "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 accelerators, antioxidants, binders, buffers, carriers, coating agents, colorants, diluents, disintegrants, emulsifiers, extenders, fillers, flavorings, humectants, lubricants, fragrances, preservatives, propellants, release agents, sterilants, sweeteners, solubilizers, wetting agents and mixtures thereof. The term "excipient" may also refer to a diluent, adjuvant (e.g., Freund's adjuvant) (complete or incomplete) or vehicle.

[0233] In one embodiment, each component is "pharmaceutically acceptable" in the sense that it is compatible with the other ingredients of the pharmaceutical formulation and suitable for use in contact with tissues or organs of humans and animals without excessive toxicity, irritation, allergic response, immunogenicity, or other problems or complications commensurate with a reasonable benefit / risk ratio. See, for example, 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, a pharmaceutically acceptable excipient is an aqueous pH buffered solution.

[0234] The abbreviation "MMAE" refers to monomethylauristatin E.

[0235] Unless the context indicates otherwise, a hyphen (-) indicates the point of attachment to the side molecule.

[0236] The term "chemotherapeutic agent" refers to all compounds that are effective in inhibiting tumor growth. Non-limiting examples of chemotherapeutic agents include: alkylating agents, such as nitrogen mustards, ethyleneimine compounds, and alkyl sulfonates; antimetabolites, such as folic acid, purine or pyrimidine antagonists; mitotic inhibitors, such as anti-tubulin agents, such as vinca alkaloids, auristatins, and derivatives of podophyllotoxin; 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, biomolecules, and small molecules.

[0237] As used herein, the term "conservative substitution" refers to amino acid substitutions that are known to those skilled in the art and that can generally be performed without changing the biological activity of the resulting molecule. It is recognized by those skilled in the art that, in general, single amino acid substitutions in non-essential regions of a polypeptide do not substantially change biological activity (see, for example, Watson et al., MOLECULARBIOLOGY OF THE GENE, The Benjamin / Cummings Pub. Co., p. 224 (4th edition 1987)). Such exemplary substitutions are preferably performed according to those substitutions set forth in Tables 2 and 3. For example, such changes include substitutions of any one of isoleucine (I), valine (V), and leucine (L) for any of these hydrophobic amino acids; aspartic acid (D) for glutamic acid (E) and vice versa; glutamine (Q) for asparagine (N) and vice versa; and serine (S) for threonine (T) and vice versa. Depending on the environment of a particular amino acid and its role in the three-dimensional structure of the protein, other substitutions may also be considered conservative. For example, glycine (G) and alanine (A) are often interchangeable, and alanine (A) and valine (V) are also interchangeable. Methionine (M), which is relatively hydrophobic, is often interchangeable with leucine and isoleucine, and is sometimes interchangeable with valine. Lysine (K) and arginine (R) are often interchangeable in positions where the distinguishing characteristic of the amino acid residue is its charge, and the difference in the pK of the two amino acid residues is not significant. Still other changes can be considered "conservative" in specific circumstances (see, for example, Table 3 herein; "Biochemistry," 2nd edition, Lubert Stryer, ed. (Stanford University), pp. 13-15; 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 also permissible and can be determined empirically or based on known conservative substitutions.

[0238] Table 2 Amino acid abbreviations

[0239] Single letter Three letters Full name F Phe Phenylalanine L Leu Leucine S Ser Serine Y Tyr Tyrosine C Cys Cysteine W Trp Tryptophan P Pro Proline H His Histidine Q Gln glutamine R Arg Arginine I Ile Isoleucine M Met Methionine T Thr Threonine N Asn Asparagine K Lys Lysine V Val Valine A Ala Alanine D Asp Aspartic acid E Glu glutamate G Gly Glycine

[0240] Table 3 Amino acid substitution or similarity matrix

[0241] Adapted from GCG software 9.0 BLOSUM62 amino acid substitution matrix (block substitution matrix). The higher the value, the more likely the substitution is to be found in the related natural protein.

[0242]

[0243] The term "homology" or "homologous" is intended to mean the sequence similarity between two polynucleotides or between two polypeptides. Similarity can be determined by comparing the position in each sequence compared for comparison purposes. If the given position of two polypeptide sequences is different, the similarity or conservation of the position can be determined by, for example, assessing the amino acid similarity of the position according to Table 3. The degree of similarity between the sequences varies with the number of matching or homologous positions shared by the sequences. Comparing two sequences to determine their percentage sequence similarity can be performed using a software program known in the art, such as the software program described in Ausubel et al., Current Protocols in Molecular Biology, John Wiley and Sons, Baltimore, MD (1999). Preferably, comparison is performed using default parameters, an example of which is set forth below. A well-known alignment program that can be used in the art is BLAST, which is set to default parameters. Specifically, the programs were BLASTN and BLASTP, using the following default parameters: genetic code = standard; filter = none; strand = both; cutoff = 60; expectation = 10; matrix = BLOSUM62; description = 50 sequences; sorting = 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.

[0244] The term "homolog" of a given amino acid sequence or nucleic acid sequence is intended to indicate that the corresponding sequence of the "homolog" has substantial identity or homology with the given amino acid sequence or nucleic acid sequence.

[0245] 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 for comparing two sequences is the algorithm of Karlin and Altschul, 1990, Proc. Natl. Acad. Sci. USA 87:2264-2268, as modified in Karlin and Altschul, 1993, Proc. Natl. Acad. Sci. USA 90:5873-5877. Such algorithms are incorporated into the NBLAST and XBLAST programs of Altschul et al., 1990, J. Mol. Biol. 215:403. BLAST nucleotide searches can be performed using the NBLAST nucleotide program parameter set (e.g., for score=100, wordlength=12) to obtain nucleotide sequences homologous to the nucleic acid molecules described herein. BLAST protein searches can be performed using the XBLAST program parameter set, e.g., for a score of 50 and a wordlength of 3, to obtain amino acid sequences homologous to the protein molecules described herein. To perform gapped alignments for comparison purposes, Gapped BLAST can be used 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 (supra). When utilizing BLAST, Gapped BLAST, and PSIBlast 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, 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 4:1117. 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.

[0246] The percent identity between two sequences can be determined using techniques similar to those described above, with or without allowing gaps. When calculating percent identity, typically only exact matches are counted.

[0247] The term "cytotoxic agent" refers to a substance that inhibits or prevents cell expression activity, cell function and / or causes cell destruction. The term is intended to include radioactive isotopes, chemotherapeutic agents, and toxins, such as small molecule toxins or enzymatically active toxins derived from bacteria, fungi, plants or animals, including fragments and / or variants thereof. Examples of cytotoxic agents include, but are not limited to, auristatins (e.g., auristatin E, auristatin F, MMAE and MMAF), aureomycin, maytansinoids, ricin, ricin A chain, combrestatin, duocarmycins, dolastatins, doxorubicin, daunorubicin, taxols, cisplatin, cc1065, ethidium bromide. , mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicine, dihydroxyanthracin dione, actinomycin, diphtheria toxin, Pseudomonas aeruginosa exotoxin (PE) A, PE40, abrin, abrin A chain, modeccin A chain, α-sarcin, gelonin, mitogellin, retstrictocin, phenomycin, enomycin, curicin, crotin, calicheamicin, Sapaonaria officinalis inhibitors and glucocorticoids and other chemotherapeutic agents, and radioactive isotopes such as At 211 , I 131 , I 125 、Y 90 、Re 186 、Re 188 、Sm 153 、Bi 212 Or Bi 213 、p 32 and radioactive isotopes of Lu, including Lu 177 The antibody can also be coupled to an anticancer prodrug activating enzyme that is capable of converting the prodrug into its active form.

[0248] As used herein, the term "effective amount" or "therapeutically effective amount" refers to the amount of a binding molecule (eg, antibody) or pharmaceutical composition provided herein sufficient to produce a desired result.

[0249] The terms "subject" and "patient" are used interchangeably. As used herein, in certain embodiments, the subject is a mammal, such as a non-primate (e.g., a cow, pig, horse, cat, dog, rat, etc.) or a primate (e.g., a monkey and a human). In specific embodiments, the subject is a human. In one embodiment, the subject is a mammal, such as a human, diagnosed with a disease or condition. In another embodiment, the subject is a mammal, such as a human, at risk of developing a disease or condition.

[0250] "Administer" or "administration" refers to the act of injecting or otherwise physically delivering a substance present outside the body into a patient's body, for example, by transmucosal, intradermal, intravenous, intramuscular delivery, and / or any other physical delivery method described herein or known in the art.

[0251] As used herein, the terms "treat / treatment / treating" refer to a reduction or improvement in the progression, severity, and / or duration of a disease or condition resulting from the administration of one or more therapies. Treatment can be determined by assessing whether there has been a reduction, alleviation, and / or alleviation of one or more symptoms associated with the underlying condition, such that improvement is observed in the patient, even though the patient may still be suffering from the underlying condition. The term "treat" includes both handling and ameliorating a disease. The term "manage" refers to the beneficial effects a subject obtains from a therapy, which does not necessarily result in a cure of the disease.

[0252] The terms "prevent," "preventing," and "prevention" refer to reducing the likelihood of the onset (or recurrence) of a disease, disorder, condition, or related symptoms (eg, cancer).

[0253] The term "cancer" or "cancer cell" is used herein to refer to tissues or cells found in neoplasms that possess characteristics that distinguish them from normal tissues or tissue cells. Such characteristics include, but are not limited to, the degree of anaplasia, irregularity of shape, unclear cell outlines, changes in nuclear size, nuclear or cytoplasmic structure, other phenotypic changes, the presence of cellular proteins indicative of a cancerous or precancerous state, an increased number of mitoses, and the ability to metastasize. The term "cancer" includes carcinoma, sarcoma, tumor, epithelioma, leukemia, lymphoma, polyps and scirrhosis, transformation, neoplasm, and the like.

[0254] As used herein, "locally advanced" cancer refers to cancer that has spread from where it started to nearby tissues or lymph nodes.

[0255] As used herein, "metastatic" cancer refers to cancer that has spread from where it started to different parts of the body.

[0256] 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 stated value or range.

[0257] As used in this disclosure and the claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0258] It is to be understood that when an embodiment is described herein using the term "comprising," other similar embodiments described using the terms "consisting of" and / or "consisting essentially of" are also provided. It is also to be understood that when an embodiment is described herein using the phrase "consisting essentially of," other similar embodiments described using the term "consisting of" are also provided.

[0259] The term "and / or" as used herein in phrases such as "A and / or B" is intended to include A and B; A or B; A (alone); and B (alone). Similarly, the term "and / or" as used in phrases such as "A, B and / or C" 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 (alone); B (alone); and C (alone).

[0260] The term "variant" refers to a molecule that exhibits differences from a described type or norm, such as in a particular described protein (e.g. Figure 1A Analogs are examples of variant proteins. Splice isoforms and single nucleotide polymorphisms (SNPs) are other examples of variants.

[0261] The "191P4D12 proteins" and / or "191P4D12-related proteins" of the present disclosure include those proteins specifically identified herein (see Figure 1A), as well as allelic variants, conservative substitution variants, analogs, and homologs that can be isolated / generated and characterized without undue experimentation following methods outlined herein or readily available in the art. Fusion proteins combining portions of different 191P4D12 proteins or fragments thereof, as well as fusion proteins of 191P4D12 proteins with heterologous polypeptides are also included. 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 protein having 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or more amino acids; or at least 30, 35, 40, 45, 50, 55, 60, 65, 70, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 225, 250, 275, 300, 325, 330, 335, 339 or more amino acids or a 191P4D12 protein sequence. The term "191P4D12" can be used interchangeably with connexin-4.

[0262] 5.2 Methods for treating cancer

[0263] Urothelial carcinoma and bladder cancer (including locally advanced urothelial carcinoma, metastatic urothelial carcinoma, locally advanced bladder cancer and metastatic bladder cancer) in patients who are not suitable for cisplatin are particularly difficult to treat diseases. Typically, these patients are weak, suffer from a variety of comorbidities other than their urothelial carcinoma / bladder cancer, and cannot tolerate other treatments, causing many patients to completely interrupt therapy. Therefore, these patients have poor prognosis and very few treatment options. The present disclosure is partly based on the results of the first clinical trial to show objective response, wherein the combination of enroclozumab vedotin and pembrolizumab is applied as a first-line treatment to patients with unresectable locally advanced or metastatic urothelial carcinoma (1a / mUC) who cannot accept chemotherapy based on cisplatin. The present disclosure therefore provides a method for effectively treating urothelial carcinoma and / or bladder cancer (including locally advanced urothelial carcinoma, metastatic urothelial carcinoma, locally advanced bladder cancer and metastatic bladder cancer) patients, wherein the patient cannot accept chemotherapy based on cisplatin in this case due to insufficient renal function or other conditions as provided herein. Prior to the results described herein, there was considerable uncertainty as to whether the methods provided herein would be effective, given that the history of this patient population has proven so difficult to treat. As further described below, the levels of efficacy achieved are particularly remarkable and surprising.

[0264] 5.2.1 Methods of treating cancer in general and selected patients

[0265] Provided herein are first-line and second-line treatments for a variety of cancers in subjects, including subjects with unresectable locally advanced or metastatic urothelial carcinoma (la / mUC) who are unable to receive cisplatin-based chemotherapy, using antibody drug conjugates (ADCs) that bind 191P4D12 in combination with an anti-PD-1 antibody, such as pembrolizumab.

[0266] In one aspect, provided herein is a method for treating a subject's cancer using an ADC and an anti-PD-1 antibody (e.g., pembrolizumab) in combination with 191P4D12. In some embodiments, the human subject treated with the methods provided herein has not previously received cancer treatment other than the ADC in combination with 191P4D12. In certain embodiments, the human subject treated with the methods provided herein has not previously received a treatment comprising or consisting of an immune checkpoint inhibitor (CPI). In some embodiments, the CPI is an anti-PD-1 antibody (e.g., pembrolizumab). In some embodiments, the CPI is a PD-1 inhibitor, a PD-L1 inhibitor, or a PD-L2 inhibitor (including but not limited to atezolizumab, pembrolizumab, nivolumab, durvalumab, or avelumab). In specific embodiments, the CPI is atezolizumab, pembrolizumab, nivolumab, durvalumab, or avelumab. In certain embodiments, the human subjects treated with the methods provided herein have not received an agent (including but not limited to CD137 agonists, CTLA 4 inhibitors or OX-40 agonists) for another stimulating or co-inhibitory T cell receptor. In specific embodiments, the agent for another stimulating or co-inhibitory T cell receptor is a CD137 agonist, CTLA 4 inhibitor or OX-40 agonist. In some embodiments, the human subjects treated with the methods provided herein are not suitable for receiving cisplatin treatment. In other embodiments, the human subjects treated with the methods provided herein are not suitable for receiving cisplatin treatment and have not previously received treatment including or consisting of CPIs. In certain embodiments, the human subjects treated with the methods provided herein are not suitable for receiving cisplatin treatment, have not previously received treatment including or consisting of CPIs, and have not received auxiliary / neoadjuvant based platinum therapy within the 12 months prior to random assignment. In other embodiments, the human subjects treated with the methods provided herein are not suitable for cisplatin treatment, have not previously received treatment including or consisting of a CPI, have not received adjuvant / neoadjuvant platinum-based therapy within 12 months prior to randomization, and have not previously received systemic treatment for locally advanced or metastatic disease. In some embodiments, the cancer is urothelial carcinoma. In certain embodiments, the cancer is bladder cancer. In certain embodiments, the cancer is renal pelvis cancer. In certain embodiments, the cancer is ureteral cancer. In certain embodiments, the cancer is urethral cancer. In one embodiment, the cancer is locally advanced cancer. In another embodiment, the cancer is metastatic cancer. In another embodiment, the cancer is locally advanced urothelial carcinoma. In another embodiment, the cancer is unresectable locally advanced urothelial carcinoma. In another embodiment, the cancer is metastatic urothelial carcinoma. In one embodiment, the cancer is locally advanced bladder cancer. In another embodiment, the cancer is metastatic bladder cancer.

[0267] 5.2.1.1 Patients Not Suitable for Cisplatin

[0268] For the methods provided herein, including but not limited to the methods in the preceding paragraphs, different conditions can be used to determine the cisplatin ineligibility of a human subject. In one embodiment, the conditions for determining cisplatin ineligibility include or consist of: an ECOG performance status of 2 points. In some embodiments, the conditions for determining cisplatin ineligibility include or consist of: impaired renal function (e.g., glomerular filtration rate (GFR) or creatinine clearance <60 mL / min but ≥30 mL / min (estimated according to Croft-Gault formula, modified diet in renal disease [MDRD], or 24-hour urine)). In certain embodiments, the conditions for determining cisplatin ineligibility include or consist of: hearing loss of no less than grade 2 (e.g., hearing loss of ≥ grade 2 according to CI CTCAE version 4.03). In certain embodiments, the conditions for determining cisplatin ineligibility include or consist of: NYHA class III heart failure. In one embodiment, the conditions for determining cisplatin ineligibility include or consist of: an ECOG performance status of 2 points and impaired renal function. In some embodiments, the conditions for determining cisplatin ineligibility comprise or consist of an ECOG performance status of 2 and a hearing loss of no less than grade 2. In other embodiments, the conditions for determining cisplatin ineligibility comprise or consist of impaired renal function and a hearing loss of no less than grade 2. In other embodiments, the conditions for determining cisplatin ineligibility comprise or consist of impaired renal function and NYHA class III heart failure. In other embodiments, the conditions for determining cisplatin ineligibility comprise or consist of an ECOG performance status of 2, impaired renal function, and a hearing loss of no less than grade 2. In some embodiments, the conditions for determining cisplatin ineligibility comprise or consist of impaired renal function, a hearing loss of no less than grade 2, and NYHA class III heart failure. In some embodiments, the conditions for determining cisplatin ineligibility comprise or consist of any one of an ECOG performance status of 2, impaired renal function, and a hearing loss of no less than grade 2. In some embodiments, the conditions for determining cisplatin ineligibility comprise or consist of any one of impaired renal function, hearing loss of grade 2 or greater, and NYHA class III heart failure. In some embodiments, the conditions for determining cisplatin ineligibility comprise or consist of any combination or permutation of any two of an ECOG performance status of 2, impaired renal function, and hearing loss of grade 2 or greater. In some embodiments, the conditions for determining cisplatin ineligibility comprise or consist of any combination or permutation of any two of impaired renal function, hearing loss of grade 2 or greater, and NYHA class III heart failure.In some embodiments, the conditions for determining cisplatin ineligibility include or consist of: an ECOG performance status of 2, impaired renal function, and hearing loss of no less than grade 2. In some embodiments, the conditions for determining cisplatin ineligibility include or consist of: all three of impaired renal function, hearing loss of no less than grade 2, and NYHA class III heart failure. In some embodiments of the methods provided herein, the subject has not previously received systemic treatment for locally advanced or metastatic disease. In some embodiments of the methods provided herein, the subject has not received adjuvant / neoadjuvant platinum-based therapy within 12 months prior to randomization. In some embodiments of the methods provided herein, wherein the subject has an ECOG performance status of 2, the subject (i) has hemoglobin ≥ 10 g / dL; (ii) has a GFR ≥ 50 mL / min; and (iii) does not have NYHA class III heart failure.

[0269] Impaired renal function can be determined using different methods known and available in the art. For the methods provided herein, including but not limited to the methods in the aforementioned paragraphs, various embodiments are provided herein to determine the impaired renal function of human subjects. In one embodiment, impaired renal function is determined based on a glomerular filtration rate (GFR) of less than 60 mL / min. In some embodiments, impaired renal function is determined based on a GFR of less than 60 but not less than 30 mL / min. In certain embodiments, impaired renal function is determined based on a GFR of less than 30 but not less than 15 mL / min. In some embodiments of the method provided in this paragraph, GFR is measured by 24-hour urine collection. In other embodiments of the method provided in this paragraph, GFR is estimated according to the Croft-Gault standard. In other embodiments of the method provided in this paragraph, GFR is measured by Modification of Diet for Renal Disease [MDRD].

[0270] In another embodiment, impaired renal function is determined based on a creatinine clearance (CrCl) of less than 60 mL / min. In some embodiments, impaired renal function is determined based on a CrCl of less than 60 but not less than 30 mL / min. In certain embodiments, impaired renal function is determined based on a CrCl of less than 30 but not less than 15 mL / min. In some embodiments of the method provided in this paragraph, CrCl is measured by 24-hour urine collection. In other embodiments of the method provided in this paragraph, CrCl is estimated according to the Croft-Gault criteria.

[0271] Thus, for the methods provided herein, including but not limited to the methods in the preceding paragraphs, certain specific conditions based on GFR or creatinine clearance can be utilized to determine cisplatin ineligibility in a human subject. In some embodiments, the conditions for determining cisplatin ineligibility include or consist of: a GFR less than 60 mL / min. In one embodiment, the conditions for determining cisplatin ineligibility include or consist of: an ECOG performance status of 2 and CrCl less than 60 mL / min. In other embodiments, the conditions for determining cisplatin ineligibility include or consist of: a GFR less than 60 mL / min and a hearing loss of no less than grade 2. In other embodiments, the conditions for determining cisplatin ineligibility include or consist of: a GFR less than 60 mL / min and NYHA class III heart failure. In other embodiments, the conditions for determining cisplatin ineligibility include or consist of: an ECOG performance status of 2, a GFR less than 60 mL / min, and a hearing loss of no less than grade 2. In other embodiments, the conditions for determining cisplatin ineligibility comprise or consist of: a GFR of less than 60 mL / min and hearing loss of no less than grade 2 and NYHA class III heart failure. In some embodiments, the conditions for determining cisplatin ineligibility comprise or consist of: any one of an ECOG performance status of 2, a GFR of less than 60 mL / min, and hearing loss of no less than grade 2. In other embodiments, the conditions for determining cisplatin ineligibility comprise or consist of: any one of a GFR of less than 60 mL / min, and hearing loss of no less than grade 2 and NYHA class III heart failure. In some embodiments, the conditions for determining cisplatin ineligibility comprise or consist of: any combination or permutation of any two of an ECOG performance status of 2, a GFR of less than 60 mL / min, and hearing loss of no less than grade 2. In other embodiments, the conditions for determining cisplatin ineligibility comprise or consist of: any combination or permutation of any two of a GFR of less than 60 mL / min, and hearing loss of no less than grade 2 and NYHA class III heart failure. In some embodiments, the conditions for determining cisplatin unsuitability include or consist of: an ECOG performance status of 2, a GFR of less than 60 mL / min, and a hearing loss of no less than grade 2. In some embodiments, the conditions for determining cisplatin unsuitability include or consist of: a GFR of less than 60 mL / min, a hearing loss of no less than grade 2, and a NYHA class III heart failure. In some embodiments of the methods provided in this paragraph, GFR is measured by 24-hour urine collection. In other embodiments of the methods provided in this paragraph, GFR is estimated according to the Croft-Gault criteria.In other embodiments of the method provided in this paragraph, GFR is measured by Modification of Diet in Renal Disease [MDRD]. In some embodiments of the method provided herein, the subject has not previously received systemic treatment for locally advanced or metastatic disease. In some embodiments of the method provided herein, the subject has not received adjuvant / neoadjuvant platinum-based therapy within 12 months prior to randomization. In some embodiments of the method provided herein, wherein the subject has an ECOG performance status of 2 points, the subject (i) hemoglobin ≥ 10 g / dL; (ii) GFR ≥ 50 mL / min; and (iii) does not have NYHA Class III heart failure.

[0272] In other embodiments, the conditions for determining cisplatin ineligibility comprise or consist of: a CrCl of less than 60 mL / min. In one embodiment, the conditions for determining cisplatin ineligibility comprise or consist of: an ECOG performance status of 2 and a CrCl of less than 60 mL / min. In other embodiments, the conditions for determining cisplatin ineligibility comprise or consist of: a CrCl of less than 60 mL / min and a hearing loss of not less than grade 2. In other embodiments, the conditions for determining cisplatin ineligibility comprise or consist of: a CrCl of less than 60 mL / min and NYHA class III heart failure. In other embodiments, the conditions for determining cisplatin ineligibility comprise or consist of: an ECOG performance status of 2, a CrCl of less than 60 mL / min and a hearing loss of not less than grade 2. In other embodiments, the conditions for determining cisplatin ineligibility comprise or consist of: a CrCl of less than 60 mL / min and a hearing loss of not less than grade 2 and NYHA class III heart failure. In some embodiments, the conditions for determining cisplatin ineligibility include or consist of: an ECOG performance status of 2, a CrCl of less than 60 mL / min, and any one of a hearing loss of grade 2 or higher. In other embodiments, the conditions for determining cisplatin ineligibility include or consist of: a CrCl of less than 60 mL / min, and any one of a hearing loss of grade 2 or higher, and NYHA Class III heart failure. In some embodiments, the conditions for determining cisplatin ineligibility include or consist of: any combination or permutation of any two of an ECOG performance status of 2, a CrCl of less than 60 mL / min, and a hearing loss of grade 2 or higher. In other embodiments, the conditions for determining cisplatin ineligibility include or consist of: any combination or permutation of any two of a CrCl of less than 60 mL / min, and any one of a hearing loss of grade 2 or higher, and NYHA Class III heart failure. In some embodiments, the conditions for determining cisplatin unsuitability include or consist of: an ECOG performance status of 2, a CrCl of less than 60 mL / min, and a hearing loss of not less than grade 2. In some embodiments, the conditions for determining cisplatin unsuitability include or consist of: a CrCl of less than 60 mL / min, a hearing loss of not less than grade 2, and a NYHA class III heart failure. In some embodiments of the method provided in this paragraph, CrCl is measured by 24-hour urine collection. In other embodiments of the method provided in this paragraph, CrCl is estimated according to the Croft-Gault criteria. In some embodiments of the method provided herein, the subject has not previously received systemic treatment for locally advanced or metastatic disease.In some embodiments of the methods provided herein, the subject has not received adjuvant / neoadjuvant platinum-based therapy within 12 months prior to randomization. In some embodiments of the methods provided herein, wherein the subject has an ECOG performance status of 2, the subject (i) has a hemoglobin ≥ 10 g / dL; (ii) has a GFR ≥ 50 mL / min; and (iii) does not have NYHA Class III heart failure.

[0273] Alternatively, for the methods provided herein, including but not limited to the methods in the preceding paragraphs, other specific conditions based on GFR or creatinine clearance can be used to determine cisplatin ineligibility in a human subject. In some embodiments, the conditions for determining cisplatin ineligibility include or consist of: a GFR less than 60 but not less than 30 mL / min. In one embodiment, the conditions for determining cisplatin ineligibility include or consist of: an ECOG performance status of 2 and a GFR less than 60 but not less than 30 mL / min. In other embodiments, the conditions for determining cisplatin ineligibility include or consist of: a GFR less than 60 but not less than 30 mL / min and a hearing loss of not less than grade 2. In other embodiments, the conditions for determining cisplatin ineligibility include or consist of: a GFR less than 60 but not less than 30 mL / min and NYHA class III heart failure. In other embodiments, the conditions for determining cisplatin ineligibility include or consist of: an ECOG performance status of 2, a GFR less than 60 but not less than 30 mL / min, and a hearing loss of not less than grade 2. In other embodiments, the conditions for determining cisplatin ineligibility comprise or consist of: a GFR less than 60 but not less than 30 mL / min, a hearing loss of not less than grade 2, and NYHA class III heart failure. In some embodiments, the conditions for determining cisplatin ineligibility comprise or consist of: an ECOG performance status of 2, a GFR less than 60 but not less than 30 mL / min, and any one of a hearing loss of not less than grade 2. In some embodiments, the conditions for determining cisplatin ineligibility comprise or consist of: a GFR less than 60 but not less than 30 mL / min, a hearing loss of not less than grade 2, and NYHA class III heart failure. In some embodiments, the conditions for determining cisplatin ineligibility comprise or consist of: any combination or permutation of any two of an ECOG performance status of 2, a GFR less than 60 but not less than 30 mL / min, and a hearing loss of not less than grade 2. In some embodiments, the conditions for determining cisplatin ineligibility comprise or consist of any combination or permutation of any two of a GFR less than 60 but not less than 30 mL / min, a hearing loss of not less than grade 2, and NYHA class III heart failure. In some embodiments, the conditions for determining cisplatin ineligibility comprise or consist of all three of an ECOG performance status of 2, a GFR less than 60 but not less than 30 mL / min, and a hearing loss of not less than grade 2. In some embodiments, the conditions for determining cisplatin ineligibility comprise or consist of all three of a GFR less than 60 but not less than 30 mL / min, a hearing loss of not less than grade 2, and NYHA class III heart failure.In some embodiments of the method provided in this paragraph, GFR is measured by 24-hour urine collection. In other embodiments of the method provided in this paragraph, GFR is estimated according to the Croft-Gault criteria. In other embodiments of the method provided in this paragraph, GFR is measured by Modification of Diet in Renal Disease [MDRD]. In some embodiments of the method provided herein, the subject has not previously received systemic treatment for locally advanced or metastatic disease. In some embodiments of the method provided herein, the subject has not received adjuvant / neoadjuvant platinum-based therapy within 12 months prior to randomization. In some embodiments of the method provided herein, wherein the subject has an ECOG performance status of 2 points, the subject (i) hemoglobin ≥ 10 g / dL; (ii) GFR ≥ 50 mL / min; and (iii) does not have NYHA class III heart failure.

[0274] In other embodiments, the conditions for determining cisplatin unsuitability include or consist of: CrCl less than 60 but not less than 30 mL / min. In one embodiment, the conditions for determining cisplatin unsuitability include or consist of: an ECOG performance status of 2 and a CrCl less than 60 but not less than 30 mL / min. In other embodiments, the conditions for determining cisplatin unsuitability include or consist of: a CrCl less than 60 but not less than 30 mL / min and a hearing loss of not less than grade 2. In other embodiments, the conditions for determining cisplatin unsuitability include or consist of: a CrCl less than 60 but not less than 30 mL / min and NYHA class III heart failure. In other embodiments, the conditions for determining cisplatin unsuitability include or consist of: an ECOG performance status of 2, a CrCl less than 60 but not less than 30 mL / min and a hearing loss of not less than grade 2. In other embodiments, the conditions for determining cisplatin ineligibility comprise or consist of: a CrCl of less than 60 but not less than 30 mL / min, a hearing loss of not less than grade 2, and NYHA class III heart failure. In some embodiments, the conditions for determining cisplatin ineligibility comprise or consist of: an ECOG performance status of 2, a CrCl of less than 60 but not less than 30 mL / min, and any one of a hearing loss of not less than grade 2. In some embodiments, the conditions for determining cisplatin ineligibility comprise or consist of: a CrCl of less than 60 but not less than 30 mL / min, a hearing loss of not less than grade 2, and NYHA class III heart failure. In some embodiments, the conditions for determining cisplatin ineligibility comprise or consist of: any combination or permutation of any two of an ECOG performance status of 2, a CrCl of less than 60 but not less than 30 mL / min, and a hearing loss of not less than grade 2. In some embodiments, the conditions for determining cisplatin ineligibility include or consist of any combination or permutation of any two of a CrCl less than 60 but not less than 30 mL / min, a hearing loss of not less than grade 2, and NYHA class III heart failure. In some embodiments, the conditions for determining cisplatin ineligibility include or consist of all three of a CrCl less than 60 but not less than 30 mL / min, and a hearing loss of not less than grade 2. In some embodiments, the conditions for determining cisplatin ineligibility include or consist of all three of a CrCl less than 60 but not less than 30 mL / min, a hearing loss of not less than grade 2, and NYHA class III heart failure. In some embodiments of the methods provided in this paragraph, CrCl is measured by 24-hour urine collection. In other embodiments of the methods provided in this paragraph, CrCl is estimated according to the Croft-Gault criteria.In some embodiments of the methods provided herein, the subject has not previously received systemic treatment for locally advanced or metastatic disease. In some embodiments of the methods provided herein, the subject has not received adjuvant / neoadjuvant platinum-based therapy within 12 months prior to randomization. In some embodiments of the methods provided herein, wherein the subject has an ECOG performance status of 2, the subject (i) has hemoglobin ≥ 10 g / dL; (ii) has a GFR ≥ 50 mL / min; and (iii) does not have NYHA Class III heart failure.

[0275] Similarly, for the methods provided herein, including but not limited to the methods in the preceding paragraphs, other specific conditions based on GFR or creatinine clearance can be used to determine cisplatin ineligibility in a human subject. In some embodiments, the conditions for determining cisplatin ineligibility include or consist of: a GFR less than 30 but not less than 15 mL / min. In one embodiment, the conditions for determining cisplatin ineligibility include or consist of: an ECOG performance status of 2 and a GFR less than 30 but not less than 15 mL / min. In other embodiments, the conditions for determining cisplatin ineligibility include or consist of: a GFR less than 30 but not less than 15 mL / min and a hearing loss of not less than grade 2. In other embodiments, the conditions for determining cisplatin ineligibility include or consist of: a GFR less than 30 but not less than 15 mL / min and NYHA class III heart failure. In other embodiments, the conditions for determining cisplatin ineligibility include or consist of: an ECOG performance status of 2, a GFR less than 30 but not less than 15 mL / min, and a hearing loss of not less than grade 2. In other embodiments, the conditions for determining cisplatin ineligibility comprise or consist of: a GFR less than 30 but not less than 15 mL / min, a hearing loss of not less than grade 2, and NYHA class III heart failure. In some embodiments, the conditions for determining cisplatin ineligibility comprise or consist of: an ECOG performance status of 2, a GFR less than 30 but not less than 15 mL / min, and any one of a hearing loss of not less than grade 2. In some embodiments, the conditions for determining cisplatin ineligibility comprise or consist of: a GFR less than 30 but not less than 15 mL / min, a hearing loss of not less than grade 2, and NYHA class III heart failure. In some embodiments, the conditions for determining cisplatin ineligibility comprise or consist of: any combination or permutation of any two of an ECOG performance status of 2, a GFR less than 30 but not less than 15 mL / min, and a hearing loss of not less than grade 2. In some embodiments, the conditions for determining cisplatin ineligibility comprise or consist of any combination or permutation of any two of a GFR less than 30 but not less than 15 mL / min, a hearing loss of not less than grade 2, and NYHA class III heart failure. In some embodiments, the conditions for determining cisplatin ineligibility comprise or consist of all three of an ECOG performance status of 2, a GFR less than 30 but not less than 15 mL / min, and a hearing loss of not less than grade 2. In some embodiments, the conditions for determining cisplatin ineligibility comprise or consist of all three of a GFR less than 30 but not less than 15 mL / min, a hearing loss of not less than grade 2, and NYHA class III heart failure.In some embodiments of the method provided in this paragraph, GFR is measured by 24-hour urine collection. In other embodiments of the method provided in this paragraph, GFR is estimated according to the Croft-Gault criteria. In other embodiments of the method provided in this paragraph, GFR is measured by Modification of Diet in Kidney Disease [MDRD]. In some embodiments of the method provided herein, the subject has not previously received systemic treatment for locally advanced or metastatic disease. In some embodiments of the method provided herein, the subject has not received adjuvant / neoadjuvant platinum-based therapy within 12 months prior to randomization. In some embodiments of the method provided herein, wherein the subject has an ECOG performance status of 2 points, the subject (i) hemoglobin ≥ 10 g / dL; (ii) GFR ≥ 50 mL / min; and (iii) does not have NYHA grade III heart failure.

[0276] In some embodiments, the conditions for determining cisplatin unsuitability include or consist of: CrCl less than 30 but not less than 15 mL / min. In one embodiment, the conditions for determining cisplatin unsuitability include or consist of: an ECOG performance status of 2 and a CrCl less than 30 but not less than 15 mL / min. In other embodiments, the conditions for determining cisplatin unsuitability include or consist of: CrCl less than 30 but not less than 15 mL / min and hearing loss of not less than grade 2. In other embodiments, the conditions for determining cisplatin unsuitability include or consist of: CrCl less than 30 but not less than 15 mL / min and NYHA class III heart failure. In other embodiments, the conditions for determining cisplatin unsuitability include or consist of: an ECOG performance status of 2, a CrCl less than 30 but not less than 15 mL / min and hearing loss of not less than grade 2. In other embodiments, the conditions for determining cisplatin ineligibility comprise or consist of: a CrCl of less than 30 but not less than 15 mL / min, a hearing loss of not less than grade 2, and NYHA class III heart failure. In some embodiments, the conditions for determining cisplatin ineligibility comprise or consist of: an ECOG performance status of 2, a CrCl of less than 30 but not less than 15 mL / min, and any one of a hearing loss of not less than grade 2. In some embodiments, the conditions for determining cisplatin ineligibility comprise or consist of: a CrCl of less than 30 but not less than 15 mL / min, a hearing loss of not less than grade 2, and NYHA class III heart failure. In some embodiments, the conditions for determining cisplatin ineligibility comprise or consist of: any combination or permutation of any two of an ECOG performance status of 2, a CrCl of less than 30 but not less than 15 mL / min, and a hearing loss of not less than grade 2. In some embodiments, the conditions for determining cisplatin ineligibility include or consist of any combination or permutation of any two of a CrCl less than 30 but not less than 15 mL / min, a hearing loss of not less than grade 2, and NYHA class III heart failure. In some embodiments, the conditions for determining cisplatin ineligibility include or consist of all three of a CrCl less than 30 but not less than 15 mL / min, and a hearing loss of not less than grade 2. In some embodiments, the conditions for determining cisplatin ineligibility include or consist of all three of a CrCl less than 30 but not less than 15 mL / min, a hearing loss of not less than grade 2, and NYHA class III heart failure. In some embodiments of the methods provided in this paragraph, CrCl is measured by 24-hour urine collection. In other embodiments of the methods provided in this paragraph, CrCl is estimated according to the Croft-Gault criteria.In some embodiments of the methods provided herein, the subject has not previously received systemic treatment for locally advanced or metastatic disease. In some embodiments of the methods provided herein, the subject has not received adjuvant / neoadjuvant platinum-based therapy within 12 months prior to randomization. In some embodiments of the methods provided herein, wherein the subject has an ECOG performance status of 2, the subject (i) has hemoglobin ≥ 10 g / dL; (ii) has a GFR ≥ 50 mL / min; and (iii) does not have NYHA Class III heart failure.

[0277] 5.2.1.2 Other patient demographics

[0278] In addition, the human subjects to which the methods provided herein can be used are human subjects with various other conditions. In one embodiment, the human subjects to which the methods provided herein are used may suffer from histologically confirmed locally advanced or metastatic urothelial (formerly known as transitional cell) cancer (e.g., bladder cancer, renal pelvis cancer, ureteral cancer, or urethral cancer). In some embodiments, the human subjects to which the methods provided herein are used may be suitable for CPI therapy. In certain embodiments, the human subjects to which the methods provided herein are used may suffer from measurable disease according to RECIST version 1.1. In other embodiments, the human subjects to which the methods provided herein are used may have lesions in the previous irradiation field, and the lesions have developed to the point where they are considered measurable. In other embodiments, the human subjects to which the methods provided herein are used may have an ECOG performance status of 0. In other embodiments, the human subjects to which the methods provided herein are used may have an ECOG performance status of 1. In other embodiments, the human subjects to which the methods provided herein are used may have an ECOG performance status of 2. In other embodiments, the human subjects used in the methods provided herein may have an ECOG performance status of 1 to 2. In other embodiments, the human subjects used in the methods provided herein may have an ECOG performance status of 1 or 2. In other embodiments, the human subjects used in the methods provided herein may have an ECOG performance status of 0 to 1. In other embodiments, the human subjects used in the methods provided herein may have an ECOG performance status of 0 or 1. In other embodiments, the human subjects used in the methods provided herein may have an ECOG performance status of 0 to 2. In other embodiments, the human subjects used in the methods provided herein may have an ECOG performance status of 0 or 2. In other embodiments, the human subjects used in the methods provided herein may have an ECOG performance status of 0, 1, or 2. In one embodiment, the human subjects used in the methods provided herein may have a life expectancy of ≥ 3 months. Some embodiments of the methods provided in this paragraph may have any combination or permutation of any of the foregoing embodiments.

[0279] In other embodiments of the methods provided herein, including the methods of the preceding paragraphs, the human subjects to whom the methods provided herein may be applied are human subjects with various other conditions. In one embodiment, the human subjects to whom the methods provided herein may be applied also have the following conditions: an absolute neutrophil count (ANC) of not less than 1500 / μL. In some embodiments, the human subjects to whom the methods provided herein may be applied also have the following conditions: a platelet count of not less than 100,000 / μL. In certain embodiments, the human subjects to whom the methods provided herein may be applied also have the following conditions: a hemoglobin of not less than 9 g / dL. In certain embodiments, the human subjects to whom the methods provided herein may be applied also have the following conditions: a hemoglobin of not less than 9 g / dL, wherein the subject is not erythropoietin-dependent and wherein the subject has not received a transfusion of packed red blood cells (pRBC) within 2 weeks prior to treatment. In certain embodiments, the human subjects to whom the methods provided herein may be applied also have the following conditions: a hemoglobin of not less than 5.6 mmol / L. In certain embodiments, the human subjects for whom the methods provided herein can be used also have a hemoglobin level of no less than 5.6 mmol / L, wherein the subject is not erythropoietin-dependent and wherein the subject has not received a transfusion of packed red blood cells (pRBCs) within 2 weeks prior to treatment. In certain embodiments, the human subjects for whom the methods provided herein can be used also have a hemoglobin level of no less than 9 g / dL or 5.6 mmol / L. In other embodiments, the human subjects for whom the methods provided herein can be used also have a serum bilirubin level of no more than 1.5 times the upper limit of normal (ULN), direct bilirubin ≤ ULN, wherein the human subject has a total bilirubin level > 1.5 x ULN, or for patients with Gilbert's disease, a serum bilirubin level of no more than 3 times ULN. In other embodiments, the human subjects for whom the methods provided herein can be used also have a CrCl level of no less than 30 mL / min. In other embodiments, the human subjects for whom the methods provided herein can be used also have a GFR of no less than 30 mL / min. In other embodiments, the human subjects for whom the methods provided herein can be used further have the following conditions: CrCl or GFR is not less than 30 mL / min. In other embodiments, the human subjects for whom the methods provided herein can be used further have the following conditions: CrCl is not less than 30 mL / min, wherein the subject's creatinine level is >1.5×institutional ULN and / or ≤1.5 ULN. In other embodiments, the human subjects for whom the methods provided herein can be used further have the following conditions: GFR is not less than 30 mL / min, wherein the subject's creatinine level is >1.5×institutional ULN and / or ≤1.5 ULN.In another embodiment, the human subjects for whom the methods provided herein can be used also have the following conditions: alanine aminotransferase (ALT) and aspartate aminotransferase (AST) are no more than 3 times ULN. In one embodiment, the human subjects for whom the methods provided herein can be used also have the following conditions: absolute neutrophil count is no less than 1500 / μL and platelet count is no less than 100,000 / μL. In some embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: absolute neutrophil count is no less than 1500 / μL and hemoglobin is no less than 9 g / dL or 5.6 mmol / L. In certain embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: absolute neutrophil count is no less than 1500 / μL and serum bilirubin is no more than 1.5 times ULN, direct bilirubin ≤ ULN, wherein the human subject has the condition that the total bilirubin level is >1.5×ULN, or for patients with Gilbert's disease, the serum bilirubin is no more than 3 times ULN. In other embodiments, the human subjects to whom the methods provided herein can be applied also have the following conditions: the absolute neutrophil count is not less than 1500 / μL and CrCl or GFR is not less than 30 mL / min. In other embodiments, the human subjects to whom the methods provided herein can be applied also have the following conditions: the absolute neutrophil count is not less than 1500 / μL and CrCl is not less than 30 mL / min, wherein the subject's creatinine level is >1.5×institutional ULN and / or ≤1.5 ULN. In some embodiments, the human subjects to whom the methods provided herein can be applied also have the following conditions: the absolute neutrophil count is not less than 1500 / μL and ALT and AST are not more than 3 times ULN. In other embodiments, the human subjects to whom the methods provided herein can be applied also have the following conditions: the platelet count is not less than 100,000 / μL and the hemoglobin is not less than 9 g / dL or 5.6 mmol / L. In one embodiment, the human subjects for whom the methods provided herein can be used also have the following conditions: platelet count is not less than 100,000 / μL and serum bilirubin is not more than 1.5 times ULN, direct bilirubin ≤ ULN, wherein the human subject has the condition of total bilirubin level> 1.5×ULN, or for patients with Gilbert's disease, serum bilirubin is not more than 3 times ILN. In some embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: platelet count is not less than 100,000 / μL and CrCl or GFR is not less than 30 mL / min. In certain embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: platelet count is not less than 100,000 / μL and ALT and AST are not more than 3 times ULN.In other embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: hemoglobin is not less than 9 g / dL or 5.6 mmol / L and serum bilirubin is not more than 1.5 times ULN, direct bilirubin ≤ ULN, wherein the human subject has a total bilirubin level> 1.5 x ULN, or for patients with Gilbert's disease, serum bilirubin is not more than 3 times ULN. In other embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: hemoglobin is not less than 9 g / dL or 5.6 mmol / L and CrCl or GFR is not less than 30 mL / min. In some embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: hemoglobin is not less than 9 g / dL or 5.6 mmol / L and ALT and AST are not more than 3 times ULN. In one embodiment, the human subjects for whom the methods provided herein can be used further have the following conditions: serum bilirubin is no more than 1.5 times ULN, direct bilirubin ≤ ULN, wherein the human subject has a total bilirubin level > 1.5 x ULN, or for patients with Gilbert's disease, serum bilirubin is no more than 3 times ULN, and CrCl is no less than 30 mL / min. In another embodiment, the human subjects for whom the methods provided herein can be used further have the following conditions: serum bilirubin is no more than 1.5 times ULN, direct bilirubin ≤ ULN, wherein the human subject has a total bilirubin level > 1.5 x ULN, or for patients with Gilbert's disease, serum bilirubin is no more than 3 times ULN, and ALT and AST are no more than 3 times ULN. In another embodiment, the human subjects for whom the methods provided herein can be used further have the following conditions: CrCl or GFR is no less than 30 mL / min and ALT and AST are no more than 3 times ULN. In other embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: an absolute neutrophil count of not less than 1500 / μL, a platelet count of not less than 100,000 / μL, and a hemoglobin of not less than 9 g / dL or 5.6 mmol / L. In other embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: an absolute neutrophil count of not less than 1500 / μL, a platelet count of not less than 100,000 / μL, and a serum bilirubin of not more than 1.5 times ULN, direct bilirubin ≤ ULN, wherein the human subject has a total bilirubin level > 1.5 x ULN, or for patients with Gilbert's disease, serum bilirubin of not more than 3 times ULN. In other embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: an absolute neutrophil count of not less than 1500 / μL, a platelet count of not less than 100,000 / μL, and a CrCl or GFR of not less than 30 mL / min.In some embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: the absolute neutrophil count is not less than 1500 / μL, the platelet count is not less than 100,000 / μL, and ALT and AST are not more than 3 times ULN. In some embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: the absolute neutrophil count is not less than 1500 / μL, the hemoglobin is not less than 9 g / dL or 5.6 mmol / L, and the serum bilirubin is not more than 1.5 times ILN or for patients with Gilbert's disease, not more than 3 times ULN. In certain embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: the absolute neutrophil count is not less than 1500 / μL, the hemoglobin is not less than 9 g / dL or 5.6 mmol / L, and CrCl or GFR is not less than 30 mL / min. In some embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: an absolute neutrophil count of not less than 1500 / μL, hemoglobin of not less than 9 g / dL or 5.6 mmol / L, and ALT and AST of not more than 3 times ULN. In other embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: an absolute neutrophil count of not less than 1500 / μL, serum bilirubin of not more than 1.5 times ULN, direct bilirubin ≤ ULN, wherein the human subject has a total bilirubin level > 1.5 × ULN, or for patients with Gilbert's disease, serum bilirubin of not more than 3 times ULN, and CrCl or GFR of not less than 30 mL / min. In some embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: absolute neutrophil count is not less than 1500 / μL, serum bilirubin is not more than 1.5 times ULN, direct bilirubin ≤ ULN, wherein the human subject has a total bilirubin level> 1.5×ULN, or for patients with Gilbert's disease, serum bilirubin is not more than 3 times ILN, and ALT and AST are not more than 3 times ULN. In some embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: absolute neutrophil count is not less than 1500 / μL, CrCl or GFR is not less than 30 mL / min, and ALT and AST are not more than 3 times ULN. In some embodiments, the human subject for whom the methods provided herein can be used also has the following conditions: platelet count is not less than 100,000 / μL, hemoglobin is not less than 9 g / dL or 5.6 mmol / L, and serum bilirubin is not more than 1.5 times ULN, direct bilirubin ≤ ULN, wherein the human subject has the condition of total bilirubin level > 1.5×ULN, or for patients with Gilbert's disease, serum bilirubin is not more than 3 times ULN.In certain embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: platelet count is not less than 100,000 / μL, hemoglobin is not less than 9 g / dL or 5.6 mmol / L, and CrCl or GFR is not less than 30 mL / min. In some embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: platelet count is not less than 100,000 / μL, hemoglobin is not less than 9 g / dL or 5.6 mmol / L, and ALT and AST are not more than 3 times ULN. In other embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: platelet count is not less than 100,000 / μL, serum bilirubin is not more than 1.5 times ULN, direct bilirubin ≤ ULN, wherein the human subject has a total bilirubin level > 1.5 x ULN, or for patients with Gilbert's disease, serum bilirubin is not more than 3 times ULN, and CrCl or GFR is not less than 30 mL / min. In other embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: platelet count is not less than 100,000 / μL, serum bilirubin is not more than 1.5 times ULN, direct bilirubin ≤ ULN, wherein the human subject has a total bilirubin level> 1.5×ULN, or for patients with Gilbert's disease, serum bilirubin is not more than 3 times ULN, and ALT and AST are not more than 3 times ULN. In some embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: platelet count is not less than 100,000 / μL, CrCl or GFR is not less than 30 mL / min, and ALT and AST are not more than 3 times ULN. In other embodiments, the human subjects for whom the methods provided herein can be used further have the following conditions: hemoglobin is not less than 9 g / dL or 5.6 mmol / L, serum bilirubin is not more than 1.5 times ULN, direct bilirubin ≤ ULN, wherein the human subject has a total bilirubin level> 1.5 x ULN, or for patients with Gilbert's disease, serum bilirubin is not more than 3 times ULN, and CrCl or GFR is not less than 30 mL / min. In other embodiments, the human subjects for whom the methods provided herein can be used further have the following conditions: hemoglobin is not less than 9 g / dL or 5.6 mmol / L, serum bilirubin is not more than 1.5 times ULN, direct bilirubin ≤ ULN, wherein the human subject has a total bilirubin level> 1.5 x ULN, or for patients with Gilbert's disease, serum bilirubin is not more than 3 times ULN, and ALT and AST are not more than 3 times ULN. In certain embodiments, human subjects for whom the methods provided herein can be used also have the following conditions: hemoglobin is no less than 9 g / dL or 5.6 mmol / L, CrCl or GFR is no less than 30 mL / min, and ALT and AST are no more than 3 times ULN.In some embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: serum bilirubin is no more than 1.5 times ULN, direct bilirubin ≤ ULN, wherein the human subject has a total bilirubin level> 1.5 x ULN, or for patients with Gilbert's disease, serum bilirubin is no more than 3 times ULN, CrCl or GFR is no less than 30 mL / min, and ALT and AST are no more than 3 times ULN. In other embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: absolute neutrophil count is no less than 1500 / μL, platelet count is no less than 100,000 / μL, hemoglobin is no less than 9 g / dL or 5.6 mmol / L, and serum bilirubin is no more than 1.5 times uLN, direct bilirubin ≤ ULN, wherein the human subject has a total bilirubin level> 1.5 x ULN, or for patients with Gilbert's disease, serum bilirubin is no more than 3 times ULN. In other embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: absolute neutrophil count is not less than 1500 / μL, platelet count is not less than 100,000 / μL, hemoglobin is not less than 9 g / dL or 5.6 mmol / L, and CrCl or GFR is not less than 30 mL / min. In some embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: absolute neutrophil count is not less than 1500 / μL, platelet count is not less than 100,000 / μL, hemoglobin is not less than 9 g / dL or 5.6 mmol / L, and ALT and AST are no more than 3 times ULN. In some embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: an absolute neutrophil count of no less than 1500 / μL, a platelet count of no less than 100,000 / μL, serum bilirubin no more than 1.5 times ULN, direct bilirubin ≤ ULN, wherein the human subject has a total bilirubin level > 1.5×ULN, or for patients with Gilbert's disease, serum bilirubin no more than 3 times ULN, and CrCl or GFR no less than 30 mL / min. In some embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: an absolute neutrophil count of no less than 1500 / μL, a platelet count no less than 100,000 / μL, serum bilirubin no more than 1.5 times ULN, direct bilirubin ≤ ULN, wherein the human subject has a total bilirubin level > 1.5×ULN, or for patients with Gilbert's disease, serum bilirubin no more than 3 times ULN, and ALT and AST no more than 3 times ULN.In some embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: absolute neutrophil count is not less than 1500 / μL, platelet count is not less than 100,000 / μL, CrCl or GFR is not less than 30 mL / min, and ALT and AST are not more than 3 times ULN. In certain embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: absolute neutrophil count is not less than 1500 / μL, hemoglobin is not less than 9 g / dL or 5.6 mmol / L, serum bilirubin is not more than 1.5 times ULN, direct bilirubin ≤ ULN, wherein the human subject has a total bilirubin level> 1.5×ULN, or for patients with Gilbert's disease, serum bilirubin is not more than 3 times ULN, and CrCl or GFR is not less than 30 mL / min. In certain embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: an absolute neutrophil count of not less than 1500 / μL, hemoglobin of not less than 9 g / dL or 5.6 mmol / L, serum bilirubin of not more than 1.5 times ULN, direct bilirubin ≤ ULN, wherein the human subject has a total bilirubin level > 1.5 x ULN, or for patients with Gilbert's disease, serum bilirubin of not more than 3 times ULN, and ALT and AST of not more than 3 times ULN. In certain embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: an absolute neutrophil count of not less than 1500 / μL, hemoglobin of not less than 9 g / dL or 5.6 mmol / L, CrCl or GFR of not less than 30 mL / min, and ALT and AST of not more than 3 times ULN. In other embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: an absolute neutrophil count of not less than 1500 / μL, serum bilirubin of not more than 1.5 times ULN, direct bilirubin ≤ ULN, wherein the human subject has a total bilirubin level> 1.5×ULN, or for patients with Gilbert's disease, serum bilirubin of not more than 3 times ULN, CrCl or GFR of not less than 30 mL / min, and ALT and AST of not more than 3 times ULN. In other embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: a platelet count of not less than 100,000 / μL, hemoglobin of not less than 9 g / dL or 5.6 mmol / L, serum bilirubin of not more than 1.5 times ULN, direct bilirubin ≤ ULN, wherein the human subject has a total bilirubin level> 1.5×ULN, or for patients with Gilbert's disease, serum bilirubin of not more than 3 times ULN, and CrCl or GFR of not less than 30 mL / min.In some embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: platelet count is not less than 100,000 / μL, hemoglobin is not less than 9 g / dL or 5.6 mmol / L, serum bilirubin is not more than 1.5 times ULN, direct bilirubin ≤ ULN, wherein the human subject has a total bilirubin level> 1.5×ULN, or for patients with Gilbert's disease, serum bilirubin is not more than 3 times ULN, and ALT and AST are not more than 3 times ULN. In other embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: platelet count is not less than 100,000 / μL, hemoglobin is not less than 9 g / dL or 5.6 mmol / L, CrCl or GFR is not less than 30 mL / min, and ALT and AST are not more than 3 times ULN. In some embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: platelet count is not less than 100,000 / μL, serum bilirubin is not more than 1.5 times ULN, direct bilirubin ≤ ULN, wherein the human subject has a total bilirubin level> 1.5×ULN, or for patients with Gilbert's disease, serum bilirubin is not more than 3 times ULN, CrCl or GFR is not less than 30 mL / min, and ALT and AST are not more than 3 times ULN. In some embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: hemoglobin is not less than 9 g / dL or 5.6 mmol / L, serum bilirubin is not more than 1.5 times ULN, direct bilirubin ≤ ULN, wherein the human subject has a total bilirubin level> 1.5×ULN, or for patients with Gilbert's disease, serum bilirubin is not more than 3 times ULN, CrCl or GFR is not less than 30 mL / min, and ALT and AST are not more than 3 times ULN. In certain embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: an absolute neutrophil count of no less than 1500 / μL, a platelet count of no less than 100,000 / μL, a hemoglobin of no less than 9 g / dL or 5.6 mmol / L, a serum bilirubin of no more than 1.5 times ULN, direct bilirubin ≤ ULN, wherein the human subject has a total bilirubin level > 1.5 x ULN, or for patients with Gilbert's disease, the serum bilirubin does not exceed 3 times ULN, and CrCl or GFR is no less than 30 mL / min.In some embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: an absolute neutrophil count of not less than 1500 / μL, a platelet count of not less than 100,000 / μL, hemoglobin of not less than 9 g / dL or 5.6 mmol / L, serum bilirubin of not more than 1.5 times ULN, direct bilirubin ≤ ULN, wherein the human subject has a total bilirubin level > 1.5×ULN, or for patients with Gilbert's disease, serum bilirubin of not more than 3 times ULN, and ALT and AST of not more than 3 times ULN. In some embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: an absolute neutrophil count of not less than 1500 / μL, a platelet count of not less than 100,000 / μL, hemoglobin of not less than 9 g / dL or 5.6 mmol / L, CrCl or GFR of not less than 30 mL / min, and ALT and AST of not more than 3 times ULN. In certain embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: an absolute neutrophil count of no less than 1500 / μL, a platelet count of no less than 100,000 / μL, a serum bilirubin of no more than 1.5 times the ILN, and direct bilirubin ≤ ULN, wherein the human subject has a total bilirubin level > 1.5×ULN, or for patients with Gilbert's disease, serum bilirubin of no more than 3 times the ILN, CrCl or GFR of no less than 30 mL / min, and ALT and AST of no more than 3 times the ULN. In certain embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: absolute neutrophil count is not less than 1500 / μL, hemoglobin is not less than 9 g / dL or 5.6 mmol / L, serum bilirubin is not more than 1.5 times ULN, direct bilirubin ≤ ULN, wherein the human subject has the condition of total bilirubin level > 1.5×ULN, or for patients with Gilbert's disease, serum bilirubin is not more than 3 times ULN, CrCl or GFR is not less than 30 mL / min, and ALT and AST are not more than 3 times ULN. In some embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: platelet count is not less than 100,000 / μL, hemoglobin is not less than 9 g / dL or 5.6 mmol / L, serum bilirubin is not more than 1.5 times ILN, direct bilirubin ≤ ULN, wherein the human subject has the condition of total bilirubin level > 1.5×ULN, or for patients with Gilbert's disease, serum bilirubin is not more than 3 times ILN, CrCl or GFR is not less than 30 mL / min, and ALT and AST are not more than 3 times ULN.In some embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: absolute neutrophil count is not less than 1500 / μL, platelet count is not less than 100,000 / μL, hemoglobin is not less than 9 g / dL or 5.6 mmol / L, serum bilirubin is not more than 1.5 times ULN, direct bilirubin ≤ ULN, wherein the human subject has the condition of total bilirubin level > 1.5×ILN, or for patients with Gilbert's disease, serum bilirubin is not more than 3 times ILN, CrCl or GFR is not less than 30 mL / min, and ALT and AST are not more than 3 times ULN. In some embodiments, the human subjects for whom the methods provided herein can be used also have the following conditions: absolute neutrophil count is not less than 1500 / μL, platelet count is not less than 100,000 / μL, hemoglobin is not less than 9 g / dL or 5.6 mmol / L, serum bilirubin is not more than 1.5 times ILN, direct bilirubin ≤ ILN, wherein the human subject has the condition of total bilirubin level >1.5×ULN, or for patients with Gilbert's disease, serum bilirubin is not more than 3 times ULN, CrCl or GFR is not less than 30 mL / min, and ALT and AST are not more than 3 times ULN. In some embodiments, the human subjects for whom the methods provided herein can be used further have the following conditions: absolute neutrophil count no less than 1500 / μL, platelet count no less than 100,000 / μL, hemoglobin no less than 9 g / dL or 5.6 mmol / L, serum bilirubin no more than 1.5 times ULN, direct bilirubin ≤ ULN, wherein the human subject has the condition of total bilirubin level > 1.5×ILN, or for patients with Gilbert's disease, serum bilirubin no more than 3 times ILN, CrCl or GFR no less than 30 mL / min, and any combination or permutation of any two of ALT and AST no more than 3 times ULN. In some embodiments, the human subjects for whom the methods provided herein can be used also have any combination or permutation of any three of the following: an absolute neutrophil count of no less than 1500 / μL, a platelet count of no less than 100,000 / μL, a hemoglobin of no less than 9 g / dL or 5.6 mmol / L, a serum bilirubin of no more than 1.5 times the ULN or, for patients with Gilbert's disease, no more than 3 times the ULN, a CrCl or GFR of no less than 30 mL / min, and an ALT and AST of no more than 3 times the ULN.In some embodiments, the human subjects for whom the methods provided herein can be used further have the following conditions: absolute neutrophil count no less than 1500 / μL, platelet count no less than 100,000 / μL, hemoglobin no less than 9 g / dL or 5.6 mmol / L, serum bilirubin no more than 1.5 times ULN, direct bilirubin ≤ ULN, wherein the human subject has the condition of total bilirubin level > 1.5×ULN, or for patients with Gilbert's disease, serum bilirubin no more than 3 times ULN, CrCl or GFR no less than 30 mL / min, and ALT and AST no more than 3 times ULN in any combination or permutation. In some embodiments, the human subjects for whom the methods provided herein can be used further have the following conditions: absolute neutrophil count no less than 1500 / μL, platelet count no less than 100,000 / μL, hemoglobin no less than 9 g / dL or 5.6 mmol / L, serum bilirubin no more than 1.5 times ULN, direct bilirubin ≤ ILN, wherein the human subject has the condition of total bilirubin level > 1.5×ULN, or for patients with Gilbert's disease, serum bilirubin no more than 3 times ULN, CrCl or GFR no less than 30 mL / min, and ALT and AST no more than 3 times ULN in any combination or permutation. In some embodiments, the human subjects for whom the methods provided herein can be used further have the following conditions: absolute neutrophil count no less than 1500 / μL, platelet count no less than 100,000 / μL, hemoglobin no less than 9 g / dL or 5.6 mmol / L, serum bilirubin no more than 1.5 times ULN, direct bilirubin ≤ ULN, wherein the human subject has the condition of total bilirubin level > 1.5×ULN, or for patients with Gilbert's disease, serum bilirubin no more than 3 times ULN, CrCl or GFR no less than 30 mL / min, and ALT and AST no more than all six of 3 times ULN.

[0280] In certain embodiments, the methods provided herein are used in human subjects who have been previously treated for prostate cancer by surgery or radiation therapy at least 1 year prior to treatment with any of the methods provided herein, wherein the subject does not have prostate cancer and wherein (i) if the subject has undergone or received radical prostatectomy, the subject must have had an undetectable PSA > 1 year prior to treatment with any of the methods provided herein, (ii) if the subject has received radiation, the subject's PSA doubling time is > 1 year (based on at least 3 values ​​determined at intervals > 1 month) and the total PSA value does not meet the Phoenix criteria for biochemical recurrence (e.g., < 2.0 ng / mL above the nadir). In certain embodiments, the methods provided herein are used in human subjects who have untreated low-risk prostate cancer, wherein the subject has a Gleason score ≤ 6, is on active surveillance, and has a PSA doubling time > 1 year (based on at least 3 values ​​determined at intervals > 1 month).

[0281] In other embodiments of the methods provided herein, including the methods of the aforementioned paragraphs, the human subjects to whom the methods provided herein may be applied are human subjects without certain conditions. In one embodiment, the human subjects for whom the methods provided herein are applied have not received any CPI treatments before. CPIs are defined as PD-1 inhibitors, PD-L1 inhibitors, or PD-L2 inhibitors (including but not limited to atezolizumab, pembrolizumab, nivolumab, durvalumab, or avelumab). In some embodiments, the human subjects for whom the methods provided herein are applied have not received PD-1 inhibitors, PD-L1 inhibitors, or PD-L2 inhibitors. In certain embodiments, the human subjects for whom the methods provided herein are applied have not received atezolizumab, pembrolizumab, nivolumab, durvalumab, or avelumab. In certain embodiments, the human subjects for whom the methods provided herein are applied have not received any treatment with agents (including but not limited to CD137 agonists, CTLA 4 inhibitors, or OX-40 agonists) directed against another stimulatory or co-inhibitory T cell receptor. In certain embodiments, the human subjects to which the methods provided herein are applied have not previously been treated with any CD137 agonists, CTLA 4 inhibitors, or OX-40 agonists. In one embodiment, the human subjects to which the methods provided herein are applied may not have sensory or motor neuropathy exceeding grade 2. In one embodiment, the human subjects to which the methods provided herein are applied may not have active central nervous system metastases. In one embodiment, the human subjects to which the methods provided herein are applied may not have uncontrolled diabetes. In one embodiment, the human subjects to which the methods provided herein are applied may not have sensory or motor neuropathy exceeding grade 2 and active central nervous system metastases. In some embodiments, the human subjects to which the methods provided herein are applied may not have sensory or motor neuropathy exceeding grade 2 and uncontrolled diabetes. In other embodiments, the human subjects to which the methods provided herein are applied may not have sensory or motor neuropathy exceeding grade 2, active central nervous system metastases, and uncontrolled diabetes. In some embodiments, the human subjects for which the methods provided herein are used may not have sensory or motor neuropathy exceeding Grade 2, not have active central nervous system metastases, and not have uncontrolled diabetes. In some embodiments, the human subjects for which the methods provided herein are used may have any two of the following: not having sensory or motor neuropathy exceeding Grade 2, not having active central nervous system metastases, and not having uncontrolled diabetes in any combination or permutation. In some embodiments, the human subjects for which the methods provided herein are used may have all three of the following: not having sensory or motor neuropathy exceeding Grade 2, not having active central nervous system metastases, and not having uncontrolled diabetes.In one embodiment of the method provided in this paragraph, uncontrolled diabetes is determined by a hemoglobin A1c (HbA1c) of not less than 8%. In some embodiments of the method provided in this paragraph, uncontrolled diabetes is determined by an HbA1c between 7% and 8% and accompanied by related diabetes symptoms not otherwise specified. In other embodiments of the method provided in this paragraph, the related diabetes symptoms comprise or consist of polyuria. In some other embodiments of the method provided in this paragraph, the related diabetes symptoms comprise or consist of polydipsia. In other embodiments of the method provided in this paragraph, the related diabetes symptoms comprise or consist of both polyuria and polydipsia.

[0282] In certain embodiments, the human subjects for whom the methods provided herein are used may not have persistent clinically significant toxicity (grade 2 or higher) associated with a previous treatment. In certain embodiments, the human subjects for whom the methods provided herein are used may not have persistent clinically significant toxicity (grade 2 or higher) associated with a previous treatment, wherein the previous treatment was radiation therapy or surgery. In one embodiment, the human subjects for whom the methods provided herein are used may not have a condition requiring high-dose steroids (e.g., >10 mg / day of prednisone or equivalent) or other immunosuppressive drugs. In one embodiment, the human subjects for whom the methods provided herein are used may not have a condition requiring high-dose steroids or other immunosuppressive drugs, wherein the steroids or other immunosuppressive drugs are not inhaled or topical steroids. In certain embodiments, the human subjects for whom the methods provided herein are used may not have been treated for urothelial carcinoma with enroclozumab or other MMAE-based ADCs. In certain embodiments, the human subjects for whom the methods provided herein are used may not have a history of another aggressive malignancy within 3 years prior to treatment with the methods provided herein. In certain embodiments, the human subjects for whom the methods provided herein are used may not have evidence of residual disease of a previously diagnosed malignancy. In certain embodiments, when taking enrobinzumab vedotin for the first time, the human subjects for whom the methods provided herein are used may not have received systemic antimicrobial therapy for an active infection, wherein the infection is a viral, bacterial, or fungal infection. When taking enrobinzumab vedotin for the first time, systemic antimicrobial therapy for an active infection (viral, bacterial, or fungal). In certain embodiments, the human subjects for whom the methods provided herein are used may not have positive hepatitis B surface antigen and / or anti-hepatitis B core antibodies. In certain embodiments, the human subjects for whom the methods provided herein are used may not have active hepatitis C infection or known human immunodeficiency virus (HIV) infection. In certain embodiments, the human subjects for whom the methods provided herein are used may not have active tuberculosis. In certain embodiments, the human subjects for whom the methods provided herein are used may not have a documented history of cerebrovascular events (e.g., stroke or transient ischemic attack), unstable angina, myocardial infarction, or cardiac symptoms consistent with NYHA class IV (including congestive heart failure) within 6 months prior to the first administration of enroku. In certain embodiments, the human subjects for whom the methods provided herein are used may not have received radiation therapy or major surgery within 2 weeks prior to treatment with the methods provided herein. In certain embodiments, the human subjects for whom the methods provided herein are used may not have completed chemotherapy, biologics, or investigational drug treatment within 4 weeks prior to treatment with the methods provided herein.In certain embodiments, the human subjects used in the methods provided herein may not have a known severe (≥ Grade 3) hypersensitivity reaction to enroku or any excipients contained in the enroku pharmaceutical formulations (including histidine, trehalose dihydrate, and polysorbate 20). In certain embodiments, the human subjects used in the methods provided herein may not have a known severe (≥ Grade 3) hypersensitivity reaction to pembrolizumab or any excipients contained in the pembrolizumab pharmaceutical formulations. In certain embodiments, the human subjects used in the methods provided herein may not have active keratitis or corneal ulcers. In certain embodiments, the human subjects used in the methods provided herein may not have an active autoimmune disease requiring systemic treatment (e.g., use of disease-modifying agents, corticosteroids, or immunosuppressive drugs) within the past two years. In certain embodiments, the human subjects used in the methods provided herein may not have an active autoimmune disease requiring systemic treatment within the past two years, wherein the systemic treatment comprises a disease-modifying agent, corticosteroids, or immunosuppressive drugs. In some embodiments, the systemic treatment is not replacement therapy (e.g., thyroxine, insulin, or physiological corticosteroid replacement therapy for the treatment of adrenal or pituitary insufficiency). In some embodiments, the systemic treatment is not thyroxine, insulin, or physiological corticosteroid replacement therapy for the treatment of adrenal or pituitary insufficiency. In certain embodiments, the human subjects used for the methods provided herein may not have a history of idiopathic pulmonary fibrosis, organizing pneumonia, drug-induced pneumonia, idiopathic pneumonia, or no evidence of active pneumonia on a screening chest CT scan. In certain embodiments, the human subjects used for the methods provided herein may not have previously undergone an allogeneic stem cell or solid organ transplant. In certain embodiments, the human subjects used for the methods provided herein may not have received a live attenuated vaccine within 30 days prior to treatment with the methods provided herein. In specific embodiments, the live vaccine is a measles, mumps, rubella, varicella / zoster (chickenpox), yellow fever, rabies, BCG, or typhoid vaccine. In specific embodiments, the live attenuated vaccine is an intranasal influenza vaccine. In certain embodiments, the human subjects for whom the methods provided herein are used may not have an underlying medical condition that would impair the subject's ability to undergo or tolerate the methods provided herein.

[0283] In some embodiments of the methods provided herein, CrCl is measured by 24-hour urine collection. In other embodiments of the methods provided herein, CrCl is estimated according to the Croft-Gault criteria.

[0284] In some embodiments of the methods provided herein, GFR is measured by 24-hour urine collection. In other embodiments of the methods provided herein, GFR is estimated according to the Croft-Gault criteria. In other embodiments of the methods provided in this paragraph, GFR is measured by Modification of Diet in Renal Disease [MDRD].

[0285] In some embodiments of the methods provided herein, the subject has been treated with one or more other cancer therapies. In certain embodiments of the methods provided herein, urothelial carcinoma, including locally advanced or metastatic urothelial carcinoma, has been treated with one or more other cancer therapies.

[0286] In some embodiments, a CPI described herein may comprise or consist of any of the CPIs described in this section (Section 5.2.1).

[0287] In all methods provided herein and those methods specifically described in the preceding paragraphs: ADCs that can be used are described in Sections 3, 5.2, 5.3, 5.4, 5.5, and 6, selecting patients for treatment is exemplified herein and in this section (Section 5.2) and 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, 5.6, 5.7, and 6 below, and methods that can be used to identify therapeutic agents, select patients, determine the results of these methods are described herein and exemplified in this section (Section 5.2, including 5.2.1 and 5.2.2) and Section 6. And / or in any way serve as a biomarker for criteria of these methods, biomarkers can be determined as described in Section 5.8 or as known in the art, the treatment outcomes of the methods provided herein are described in this section (Section 5.2, including Section 5.2.1.4) and Sections 3 and 6, other treatment outcomes of the methods provided herein can be improvements of the biomarkers described herein, such as those described and exemplified in this section (Section 5.2, including 5.2.2) and Sections 3 and 6, and combination therapies including ADCs and other therapeutic agents are described in this section (Section 5.2) and Section 5.5. Therefore, it will be understood by those skilled in the art 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.

[0288] In certain embodiments, the methods provided herein are used to treat a subject with urothelial cancer that expresses 191P4D12 RNA, 191P4D12 protein, or both. In one embodiment, the methods provided herein are used to treat a subject with urothelial cancer that expresses 191P4D12 RNA, 191P4D12 protein, or both.

[0289] In certain embodiments, the methods provided herein are used to treat a subject with locally advanced urothelial carcinoma that expresses 191P4D12 RNA, 191P4D12 protein, or both. In one embodiment, the methods provided herein are used to treat a subject with locally advanced urothelial carcinoma that expresses 191P4D12 RNA, 191P4D12 protein, or both.

[0290] In certain embodiments, the methods provided herein are used to treat a subject having metastatic cancer that expresses 191P4D12 RNA, 191P4D12 protein, or both. In one embodiment, the methods provided herein are used to treat a subject having metastatic urothelial carcinoma that expresses 191P4D12 RNA, 191P4D12 protein, or both.

[0291] In some embodiments, 191P4D12 RNA expression in cancer is measured by polynucleotide hybridization, sequencing (assessing the relative abundance of the sequence) and / or PCR (including RT-PCR). In some embodiments, 191P4D12 protein expression in cancer is measured by IHC, fluorescence activated cell sorting (FACS) analysis and / or western blotting. In some embodiments, 191P4D12 protein expression in cancer is measured by more than one method. In some embodiments, 191P4D12 protein expression in cancer is measured by two IHC methods.

[0292] In some embodiments, locally advanced or metastatic urothelial carcinoma is confirmed histologically, cytologically, or both histologically and cytologically. In some embodiments, locally advanced or metastatic bladder cancer is confirmed histologically, cytologically, or both histologically and cytologically.

[0293] In some embodiments, the subject has visceral metastases. In some embodiments, the subject has lymph node-only disease. In some embodiments, the site of disease origin is in the upper urinary tract. In some embodiments, the site of disease origin is in the lower urinary tract.

[0294] In some embodiments, the subject has a PD-L1 expression combined positive score (CPS) greater than or equal to 10. PD-L1 protein expression is determined by using a combined positive score (CPS), i.e., the number of PD-L1 positive cells (i.e., tumor cells, lymphocytes, and macrophages) divided by the total number of viable tumor cells and then multiplied by 100. In some embodiments, the subject has a PD-L1 expression CPS less than 10. The connexin H-score ranges from 0 to 300, based on the following calculation: H-score = [(0 × negative cell %) + (1 × weakly positive cell %) + (2 × moderately positive cell %) + (3 × strongly positive cell %)]. The median H score for urothelial carcinoma is generally 260-270. In some embodiments, the subject has a connexin-4 H-score between 0 and 300. In some embodiments, the subject has a connexin-4 H-score between 0 and 250, between 0 and 200, between 0 and 150, between 0 and 100, or between 0 and 50. In some embodiments, the subject has a Connexin-4H-Fraction between 0 and 200.

[0295] 5.2.1.3 Checkpoint Inhibitors and Combination Therapy with Pembrolizumab

[0296] Checkpoint inhibitors

[0297] In some embodiments, the subject that can be treated with the methods provided herein has certain phenotypic or genotypic characteristics. In some embodiments, the subject has any permutation and combination of the phenotypic or genotypic characteristics described herein.

[0298] In some embodiments, the phenotypic or genotypic characteristics are determined histologically, cytologically, or both histologically and cytologically. In some embodiments of the methods provided herein, the phenotypic and / or genotypic characteristics are determined histologically and / or cytologically, based on the most recently analyzed tissue, as described in the American Society of Clinical Oncology / College of American Pathologists (ASCO / CAP) criteria, which are incorporated herein by reference in their entirety. In some embodiments, the phenotypic or genotypic characteristics are determined by sequencing, Southern hybridization, and / or Northern hybridization, including next-generation sequencing (e.g., NGS from Illumina, Inc).

[0299] In various aspects or embodiments of the methods provided herein (including the methods provided in this section (Section 5.2), such as the methods provided in this paragraph and the preceding paragraphs), the human subjects of the methods provided herein have not previously received any CPI treatment. A CPI is defined as a PD-1 inhibitor, a PD-L1 inhibitor, or a PD-L2 inhibitor (including but not limited to atezolizumab, pembrolizumab, nivolumab, durvalumab, or avelumab). In some embodiments, the human subjects for whom the methods provided herein are used have not previously received treatment with a PD-1 inhibitor, a PD-L1 inhibitor, or a PD-L2 inhibitor. In certain embodiments, the human subjects for whom the methods provided herein are used have not previously received treatment with atezolizumab, pembrolizumab, nivolumab, durvalumab, or avelumab. As used herein, the term "immune checkpoint inhibitor" or "checkpoint inhibitor" (CPI) refers to a molecule that completely or partially reduces, inhibits, interferes with, or regulates one or more checkpoint proteins. A variety of checkpoint proteins are known, such as 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 responsible for the co-stimulatory or inhibitory interactions of T cell responses. Immune checkpoint proteins appear to regulate and maintain self-tolerance and the duration and amplitude of physiological immune responses. Immune checkpoint inhibitors include antibodies or are derived from antibodies.

[0300] In certain embodiments, the check point inhibitors used in the methods provided herein may be inhibitors or activators for check point proteins that are upregulated in cancer. In some specific embodiments, the check point inhibitors used in the methods provided herein may be inhibitors or activators for check point proteins, and the check point 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. In some embodiments, the checkpoint inhibitors for the methods provided herein may be inhibitors or activators selected from the group consisting of: PD-1 inhibitors, PD-L1 inhibitors, PD-L2 inhibitors, CTLA-4 inhibitors, LAG-3 inhibitors, B7 inhibitors, TIM3 (HAVCR2) inhibitors, OX40 (CD134) inhibitors, GITR agonists, CD137 agonists, or CD40 agonists, VTCN1 inhibitors, IDO1 inhibitors, CD276 inhibitors, PVRIG inhibitors, TIGIT inhibitors, CD25 (IL2RA) inhibitors, IFNAR2 inhibitors, IFNAR1 inhibitors, CSF1R inhibitors, VSIR (VISTA) inhibitors, or therapeutic agents targeting HLA. Such inhibitors, activators, or therapeutic agents are further provided below.

[0301] 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. Patents 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 by reference in their entirety. 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. TM Commercially available.

[0302] 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, U.S. Patent Nos. 7,488,802, 7,943,743, 8,008,449, 8,168,757, 8,217,149, and PCT Patent Application Publication Nos. WO2003042402, WO2008156712, WO2010089411, WO2010036959, WO2011066342, WO2011159877, WO2011082400, and WO2011161699, all of which are incorporated herein by reference in their entirety.

[0303] "PD-1 antagonist" refers to any compound or biological molecule that blocks the binding of PD-L1 expressed on cancer cells to PD-1 expressed on immune cells (T cells, B cells, or natural killer T cells), and in specific embodiments, also blocks the binding of PD-L2 expressed on cancer cells to PD-1 expressed on immune cells. Alternative names or synonyms for PD-1 and its ligands include: Alternative names or synonyms for PD-1 include: PDCD1, PD1, CD279, and SLEB2; Alternative names or synonyms for PD-L1 include PDCD1L1, PDL1, B7H1, B7-4, CD274, and B7-H; and Alternative names or synonyms for PD-L2 include PDCD1L2, PDL2, B7-DC, Btdc, and CD273. In any of the methods of treatment, medicaments, and uses of the present invention for treating a human subject, the PD-1 antagonist blocks the binding of human PD-L1 to human PD-1, and in certain embodiments, blocks the binding of human PD-L1 and PD-L2 to human PD-1. The human PD-1 amino acid sequence can be found in NCBI Locus No.: NP_005009. The human PD-L1 and PD-L2 amino acid sequences can be found in NCBI Locus No.: NP_054862 and NP_079515, respectively.

[0304] In some embodiments, the checkpoint inhibitor is a PD-1 inhibitor or antagonist. In one embodiment, the PD-1 inhibitor or antagonist 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, SCH 900475, or lambrolizumab). In one embodiment, the anti-PD-1 antibody is nivolumab. Nivolumab is a human IgG4 anti-PD-1 monoclonal antibody and is marketed under the trade name Opdivo.TM In another embodiment, the anti-PD-1 antibody is pembrolizumab. Pembrolizumab is a humanized monoclonal IgG4 antibody and is marketed under the trade name Keytruda®. TM Commercially available. In another embodiment, the anti-PD-1 antibody is the humanized antibody CT-011. CT-011 alone did not show a response in the treatment of relapsed acute myeloid leukemia (AML). In 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 whose ability to bind to Fcγ receptor I is specifically engineered, and it has a unique binding tag, high affinity, and excellent target specificity for PD-1. In one embodiment, the PD-1 antibody is cemiplimab. In another embodiment, the PD-1 antibody is camrelizumab. In another 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 another embodiment, the PD-1 antibody is PDR001. In another embodiment, the PD-1 antibody is toripalimab.

[0305] 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 another embodiment, the PD-L1 inhibitor is atezolizumab (also known as MPDL3280A, and In another embodiment, the PD-L1 inhibitor is avelumab.

[0306] 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.

[0307] In one embodiment, the checkpoint inhibitor is a lymphocyte activation gene-3 (LAG-3) inhibitor. In one embodiment, the LAG-3 inhibitor is a soluble Ig fusion protein IMP321 (Brignone et al., J. Immunol., 2007, 179, 4202-4211). In another embodiment, the LAG-3 inhibitor is BMS-986016.

[0308] 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 the anti-B7-H3 antibody MGA271 (Loo et al., Clin. Cancer Res., 2012, 3834).

[0309] 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).

[0310] 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.

[0311] 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.

[0312] 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.

[0313] 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.

[0314] In one embodiment, the checkpoint inhibitor is recombinant human leukocyte inhibitory factor-15 (rhIL-15).

[0315] In one embodiment, the checkpoint inhibitor is a VTCN inhibitor. In one embodiment, the VTCN inhibitor is FPA150.

[0316] 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 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 another embodiment, the IDO inhibitor is RG70099. In one embodiment, the IDO inhibitor is IOM-E. In another embodiment, the IDO inhibitor is or IOM-D.

[0317] 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 M TIG7192A. In another embodiment, the TIGIT inhibitor is BMS-986207. In 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 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 another embodiment, the TIGIT inhibitor is AJUD008.

[0318] 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 another embodiment, the VSIR inhibitor is JNJ 61610588. In one embodiment, the VSIR inhibitor is HMBD-002.

[0319] 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.

[0320] 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.

[0321] 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.

[0322] 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 another embodiment, the CSF1R inhibitor is cabiralizumab. In one embodiment, the CSF1R inhibitor is ARRY-382. In another embodiment, the CSF1R inhibitor is BLZ945. In 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 another embodiment, the CSF1R inhibitor is JNJ-40346527. In one embodiment, the CSF1R inhibitor is PLX5622. In another embodiment, the CSF1R inhibitor is FPA008.

[0323] In some embodiments, the checkpoint inhibitor is an HLA-targeting therapeutic. In certain embodiments, the HLA-targeting therapeutic is an anti-HLA antibody. In one embodiment, the HLA-targeting therapeutic is GSK01. In another embodiment, the HLA-targeting therapeutic is IMC-C103C. In another embodiment, the HLA-targeting therapeutic is IMC-F106C. In one embodiment, the HLA-targeting therapeutic is IMC-G107C. In another embodiment, the HLA-targeting therapeutic is ABBV-184.

[0324] Where appropriate, the methods described herein can be used in combination with one or more second active agents as described herein to treat diseases described herein and understood in the art.

[0325] PD-1 antagonists and pembrolizumab

[0326] Provided herein are methods for treating various cancers of subjects using an antibody drug conjugate (ADC) in combination with 191P4D12 and pembrolizumab, including subjects with unresectable locally advanced or metastatic urothelial carcinoma (1a / mUC) who cannot receive cisplatin-based chemotherapy. In certain embodiments, treatment is first-line treatment. In other embodiments, treatment is second-line treatment.

[0327] "Pembrolizumab" (formerly known as MK-3475, SCH900475, and lambliozumab), also referred to herein as "pembro," is a humanized IgG4 mAb whose structure is described in WHO Drug Information, Vol. 27, No. 2, pp. 161-162 (2013), and comprises the heavy and light chain amino acid sequences and CDRs described in Table 4. Pembrolizumab has been approved by the U.S. FDA as KEYTRUDA TM (Merck & Co., Inc., Rahway, NJ, USA; first approved in the U.S. in 2014, updated in March 2021) as described in the prescribing information.

[0328] As used herein, "pembrolizumab variant" or "variant thereof" in relation to the pembrolizumab sequence means a monoclonal antibody comprising heavy and light chain sequences that are substantially identical to those in pembrolizumab, except that the variant positions have three, two, or one conservative amino acid substitution at positions outside the light chain CDRs and six, five, four, three, two, or one conservative amino acid substitution outside the heavy chain CDRs, e.g., the variant positions are located in the FR region or the constant region, and the C-terminal lysine residue of the heavy chain is optionally deleted. In other words, pembrolizumab and pembrolizumab variants comprise the same CDR sequences but differ from each other by having conservative amino acid substitutions at no more than three or six other positions in their full-length light and heavy chain sequences, respectively. The pembrolizumab variants are substantially identical to pembrolizumab in terms of binding affinity to PD-1 and the ability to block the binding of PD-L1 and PD-L2 to PD-1.

[0329] In one embodiment, the PD-1 antagonists useful in the treatment, medicine, and use of the present invention include monoclonal antibodies (mAbs) or antigen-binding fragments thereof that specifically bind to PD-1 or PD-L1, and preferably specifically bind to human PD-1 or human PD-L1. mAbs can be human antibodies, humanized antibodies, or chimeric antibodies, and can include human constant regions. In some embodiments, the human constant region is selected from the group consisting of IgG1, IgG2, IgG3, and IgG4 constant regions, and in some embodiments, the human constant region is an IgG1 or IgG4 constant region. In some embodiments, the antigen-binding fragment is selected from the group consisting of Fab, Fab'-SH, F(ab')2, scFv, and Fv fragments.

[0330] Examples of mAbs that bind to human PD-1 and can be used in the methods of treatment, medicaments, and uses of the present invention are described in U.S. Pat. Nos. 7,488,802, 7,521,051, 8,008,449, 8,354,509, and 8,168,757, and International Application Publication Nos. WO2004 / 004771, WO2004 / 072286, WO2004 / 056875, US2011 / 0271358, and WO2008 / 156712. Specific anti-human PD-1 mAbs that can be used as PD-1 antagonists in the methods of treatment, medicaments, and uses of the present invention include: pembrolizumab (also known as MK-3475), a humanized IgG4 mAb whose structure is described in WHO Drug Information, Vol. 27, No. 2, pp. 161-162 (2013), and comprising the heavy and light chain amino acid sequences described in Table 2; nivolumab (BMS-936558), a human IgG4 mAb whose structure is described in WHO Drug Information, Vol. Information, Vol. 27, No. 1, pp. 68-69 (2013), and comprising the heavy and light chain amino acid sequences shown in Table 2; humanized antibodies h409A11, h409A16, and h409A17, which are described in WO2008 / 156712; and AMP-514 being developed by MedImmune; simvastatin; connelizumab; sintiuzumab; tislelizumab; and toripalimab. Other anti-PD-1 antibodies contemplated for use herein include MEDI0680 (U.S. Patent No. 8,609,089), BGB-A317 (U.S. Patent Publication No. 2015 / 0079109), INCSHR1210 (SHR-1210) (PCT International Application Publication No. WO2015 / 085847), REGN-2810 (PCT International Application Publication No. WO2015 / 112800), PDR001 (PCT International Application Publication No. WO2015 / 112900), TSR-042 (ANB011) (PCT International Application Publication No. WO2014 / 179664), and STI-1110 (PCT International Application Publication No. WO2014 / 194302).

[0331] Examples of mAbs that bind to human PD-L1 and can be used in the methods of treatment, medicines, and uses of the present invention are described in US 8383796. Specific anti-human PD-L1 mAbs that can be used as PD-1 antagonists in the methods of treatment, medicines, and uses of the present invention include BMS-936559, MEDI4736, and MSB0010718C.

[0332] In some embodiments, the PD-1 antagonist is pembrolizumab (KEYTRUDA TM ; Merck & Co., Inc., Rahway, NJ, USA), nivolumab (OPDIVO TM ; Bristol-Myers Squibb Company, Princeton, NJ, USA), atezolizumab (TECENTRIQ TM ; Genentech, San Francisco, CA, USA), durvalumab (IMFINZI TM ; AstraZeneca Pharmaceuticals LP, Wilmington, DE), simvastatin (LIBTAYO TM ; Regeneron Pharmaceuticals, Tarrytown, NY, USA), avelumab (BAVENCIO TM ; Merck KGaA, Darmstadt, Germany) or dostarlimab (JEMPERLI TM ; GlaxoSmithKline LLC, Philadelphia, PA). In other embodiments, the PD-1 antagonist is pidilizumab (U.S. Pat. No. 7,332,582), AMP-514 (MedImmune LLC, Gaithersburg, MD, USA), PDR001 (U.S. Pat. No. 9,683,048), BGB-A317 (U.S. Pat. No. 8,735,553), or MGA012 (MacroGenics, Rockville, MD).

[0333] In one embodiment, the PD-1 antagonist useful in the methods of the present invention is an anti-PD-1 antibody that blocks the binding of PD-1 to PD-L1 and PD-L2. In some embodiments of the methods of treatment, medicaments, and uses of the present invention, the PD-1 antagonist is a monoclonal antibody or antigen-binding fragment thereof comprising: (a) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 of SEQ ID NOs: 24, 25, and 26, respectively, and (b) a heavy chain variable region comprising heavy chain CDR1, CDR2, and CDR3 of SEQ ID NOs: 29, 30, and 31, respectively.

[0334] In other embodiments of the methods of treatment, medicaments, and uses of the present invention, the PD-1 antagonist is a monoclonal antibody or antigen-binding fragment thereof that specifically binds to human PD-1 and comprises: (a) a heavy chain variable region comprising SEQ ID NO: 32 or a variant thereof, and (b) a light chain variable region comprising SEQ ID NO: 37 or a variant thereof. The heavy chain variable region sequence variant is identical to the reference sequence except for up to six conservative amino acid substitutions in the framework regions (i.e., outside the CDRs). The light chain variable region sequence variant is identical to the reference sequence except for up to three conservative amino acid substitutions in the framework regions (i.e., outside the CDRs).

[0335] In another embodiment of the methods of treatment, medicaments, and uses of the present invention, the PD-1 antagonist is a monoclonal antibody that specifically binds to human PD-1 and comprises: (a) a heavy chain comprising SEQ ID NO: 33 and (b) a light chain comprising SEQ ID NO: 28. In one embodiment, the PD-1 antagonist is an anti-PD-1 antibody comprising two heavy chains and two light chains, wherein the heavy chain and the light chain comprise the amino acid sequences of SEQ ID NO: 33 and SEQ ID NO: 28, respectively.

[0336] In all of the above methods of treatment, medicaments, and uses, the PD-1 antagonist inhibits the binding of PD-L1 to PD-1, and in specific embodiments, also inhibits the binding of PD-L2 to PD-1. In some embodiments of the above methods of treatment, medicaments, and uses, the PD-1 antagonist is a monoclonal antibody or an antigen-binding fragment thereof that specifically binds to PD-1 or PD-L1 and blocks the binding of PD-L1 to PD-1.

[0337] Table 4 below provides a list of amino acid sequences of exemplary anti-PD-1 mAbs for use in the treatment methods, medicines, and uses of the present invention.

[0338] Table 4 Exemplary PD-1 Antibody Sequences

[0339]

[0340]

[0341] Table 5. Other PD-1 antibodies and antigen-binding fragments that can be used in the formulations, methods, and uses provided herein.

[0342]

[0343]

[0344]

[0345] In one embodiment, the anti-PD-1 antibody or its antigen-binding fragment comprises a heavy chain constant region, such as a human constant region, such as a g1, g2, g3 or g4 human heavy chain constant region or a variant thereof. In another embodiment, the anti-PD-1 antibody or its antigen-binding fragment comprises a light chain constant region, such as a human light chain constant region, such as a lambda or kappa human light chain region or a variant thereof. For example and without limitation, the human heavy chain constant region may be g4, and the human light chain constant region may be kappa. In an alternative embodiment, the Fc region of the antibody is g4 with a Ser228Pro mutation (Schuurman, J et al., Mol. Immunol. 38: 1-8, 2001). In some embodiments, different constant domains can be attached to the humanized VL and VH regions derived from the CDRs provided herein. For example, if the specific intended use of the antibody (or fragment) of the present invention requires an altered effector function, a heavy chain constant domain other than human IgG1 can be used, or a hybrid IgG1 / IgG4 can be used. Although human IgG1 antibodies have long half-life and effector functions, such as complement activation and antibody-dependent cellular toxicity, these activities may not be applicable to all uses of antibodies. In this case, for example, a human IgG4 constant region can be used. The present invention includes the use of an anti-PD-1 antibody or antigen-binding fragment thereof comprising an IgG4 constant domain. In one embodiment, the IgG4 constant domain may be different from the natural human IgG4 constant domain (Swiss-Prot accession number P01861.1) at a position corresponding to position 228 in the EU system and position 241 in the KABAT system, wherein the native Ser108 is replaced by Pro to prevent a potential interchain disulfide bond between Cys106 and Cys109 (corresponding to positions Cys 226 and Cys 229 in the EU system and positions Cys239 and Cys 242 in the KABAT system), which may interfere with the correct intrachain disulfide bond formation. See Angal et al. (1993) Mol.Imunol.30:105. In other cases, modified IgG1 constant domains may be used, where the constant region has been modified to increase half-life or reduce effector function.

[0346] In another embodiment, the PD-1 antagonist is an antibody or antigen-binding protein having a variable light domain and / or a variable heavy domain having at least 95%, 90%, 85%, 80%, 75% or 50% sequence identity with one of the above-mentioned variable light domains or variable heavy domains, and exhibiting specific binding to PD-1. In another embodiment of the treatment method of the present invention, the PD-1 antagonist is an antibody or antigen-binding protein comprising a variable light domain and a variable heavy domain having at most 1, 2, 3, 4 or 5 or more amino acid substitutions, and exhibiting specific binding to PD-1.

[0347] In some embodiments, after administering the ADC provided herein, a checkpoint inhibitor is administered. In other embodiments, the checkpoint inhibitor is administered simultaneously with the ADC provided herein (e.g., in the same administration time period). In other embodiments, after administering the ADC provided herein, a checkpoint inhibitor is administered.

[0348] In some embodiments, the amount of a checkpoint inhibitor for use in the various methods provided herein can be determined by standard clinical techniques. In certain embodiments, the amount of a PD-1 antagonist, such as pembrolizumab, for use in the various methods is provided in Section 5.6.

[0349] In some embodiments, the subject that can be treated by the methods provided herein is a mammal. In some embodiments, the subject that can be treated by the methods provided herein is a human.

[0350] 5.2.1.4 Treatment outcomes of the methods provided herein

[0351] Although there is a poor prognosis for cisplatin-unsuitable human subjects who are frail and have multiple comorbidities in addition to urothelial carcinoma / bladder cancer as described above, the methods provided herein, including the methods described in this section (Section 5.2) and Sections 3 and 6, can provide beneficial treatment results to these cisplatin-unsuitable human subjects. In one embodiment, a human subject has a complete response after treatment with the methods provided herein. In another embodiment, a human subject has a partial response after treatment with the methods provided herein. In another embodiment, a human subject has a complete response or a partial response after treatment with the methods provided herein.

[0352] In some embodiments, the response (complete or partial response) 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 described in Table 10.

[0353] In some embodiments, human subjects and patients can be used interchangeably.Thus, it will be understood by those skilled in the art that in any of the methods provided herein, human subjects can be interchanged with patients.

[0354] Thus, the therapeutic outcome of the methods provided herein can be assessed based on any one or more of the above-mentioned response criteria.

[0355] In one embodiment, a human subject experiences a partial response after treatment with the methods provided herein. In one embodiment, a human subject experiences a partial response after treatment with the methods provided herein, wherein the partial response is defined as a reduction in the sum of the diameters of the target lesions by at least or about 30%, relative to the sum of the baseline diameters. In another embodiment, a human subject experiences a partial response after treatment with the methods provided herein, wherein the partial response is defined as a reduction in the sum of the diameters of the target lesions by at least or about 35%, relative to the sum of the baseline diameters. In another embodiment, a human subject experiences a partial response after treatment with the methods provided herein, wherein the partial response is defined as a reduction in the sum of the diameters of the target lesions by at least or about 40%, relative to the sum of the baseline diameters. In another embodiment, a human subject experiences a partial response after treatment with the methods provided herein, wherein the partial response is defined as a reduction in the sum of the diameters of the target lesions by at least or about 45%, relative to the sum of the baseline diameters. In one embodiment, a human subject experiences a partial response after treatment with the methods provided herein, wherein the partial response is defined as a reduction in the sum of the diameters of the target lesions by at least or about 50%, relative to the sum of the baseline diameters. In another embodiment, a human subject experiences a partial response after treatment with the methods provided herein, wherein the partial response is defined as a reduction in the sum of the diameters of the target lesions by at least or about 55% relative to the baseline sum of the diameters. In another embodiment, a human subject experiences a partial response after treatment with the methods provided herein, wherein the partial response is defined as a reduction in the sum of the diameters of the target lesions by at least or about 60% relative to the baseline sum of the diameters. In another embodiment, a human subject experiences a partial response after treatment with the methods provided herein, wherein the partial response is defined as a reduction in the sum of the diameters of the target lesions by at least or about 65% relative to the baseline sum of the diameters. In one embodiment, a human subject experiences a partial response after treatment with the methods provided herein, wherein the partial response is defined as a reduction in the sum of the diameters of the target lesions by at least or about 70% relative to the baseline sum of the diameters. In another embodiment, a human subject experiences a partial response after treatment with the methods provided herein, wherein the partial response is defined as a reduction in the sum of the diameters of the target lesions by at least or about 75% relative to the baseline sum of the diameters. In another embodiment, a human subject experiences a partial response after treatment by the methods provided herein, wherein the partial response is defined as a reduction in the sum of the diameters of the target lesions by at least or about 80%, relative to the sum of the baseline diameters. In another embodiment, a human subject experiences a partial response after treatment by the methods provided herein, wherein the partial response is defined as a reduction in the sum of the diameters of the target lesions by at least or about 85%, relative to the sum of the baseline diameters.In one embodiment, a human subject experiences a partial response after treatment by the methods provided herein, wherein the partial response is defined as a reduction of at least or about 90% in the sum of the diameters of the target lesions relative to the baseline sum of the diameters. In another embodiment, a human subject experiences a partial response after treatment by the methods provided herein, wherein the partial response is defined as a reduction of at least or about 95% in the sum of the diameters of the target lesions relative to the baseline sum of the diameters. In some embodiments, the diameter is measured based on the longest diameter of the lesions. In certain embodiments, the diameter is measured based on the longest diameter of the lesions in a measurement plane. In some embodiments, the diameter is measured based on the longest diameter of the lesions in a measurement plane, wherein the minimum dimension is 10 mm by CT scanning. In certain embodiments, the diameter is measured based on the longest diameter of the lesions in a measurement plane, wherein the minimum dimension is 10 mm by CT scanning and the CT slice thickness does not exceed 5 mm.

[0356] The therapeutic outcome of the method provided herein can also be evaluated based on whether the disease is stable after treatment. In one embodiment, the disease of the human subject is stable after the method treatment provided by this paper. In another embodiment, the human subject does not have progressive disease after the method treatment provided by this paper.

[0357] Alternatively, the treatment outcome can be evaluated based on complete response, partial response or stable disease for a human subject population treated by the methods provided herein, by evaluating the percentage of subjects with complete response, partial response or stable disease in the treated population. Therefore, in some embodiments, the treatment outcome or efficacy measurement is applicable to the results achieved by actually treating a subject population. In other embodiments, the treatment outcome or efficacy measurement refers to the results or efficacy that can be achieved if a human subject population is treated with a method as disclosed herein. Although the following sections discuss the actual treatment of a human subject population, it should be understood that the corresponding methods that can achieve results or efficacy measurements in a patient population are also encompassed herein. Simply put, both of the above situations apply to the following sections; for simplicity and to avoid redundancy, only one situation is described below.

[0358] In some embodiments of the methods provided herein (including but not limited to, Sections 3, 5.3, 5.8, and 6, and this section (Section 5.2)), the ADC is enroku vedotin. In certain embodiments of the methods provided herein (including but not limited to, Sections 3, 5.3, 5.8, and 6, and this section (Section 5.2)), the ADC is a biosimilar of enroku vedotin. In some embodiments of the methods provided herein (including but not limited to, Sections 3, 5.2.1.3, 5.8, and 6, and this section (Section 5.2)), the PD-1 antagonist or anti-PD-1 antibody is pembrolizumab.

[0359] In one embodiment, a human subject population is treated by the method provided herein, wherein the percentage of the subject with a complete response in the treated population is at least or about 2%. In another embodiment, a human subject population is treated by the method provided herein, wherein the percentage of the subject with a complete response in the treated population is at least or about 5%. In another embodiment, a human subject population is treated by the method provided herein, wherein the percentage of the subject with a complete response in the treated population is at least or about 10%. In another embodiment, a human subject population is treated by the method provided herein, wherein the percentage of the subject with a complete response in the treated population is at least or about 10.5%. In another embodiment, a human subject population is treated by the method provided herein, wherein the percentage of the subject with a complete response in the treated population is at least or about 15%. In one embodiment, a human subject population is treated by the method provided herein, wherein the percentage of the subject with a complete response in the treated population is at least or about 20%.

[0360] In one embodiment, the complete response rate in a subject population treated with the method is at least or about 10%. In another embodiment, the complete response rate in a subject population treated with the method is at least or about 2%. In another embodiment, the complete response rate in a subject population treated with the method is at least or about 5%. In another embodiment, the complete response rate in a subject population treated with the method is at least or about 10%. In another embodiment, the complete response rate in a subject population treated with the method is at least or about 10.5%. In another embodiment, the complete response rate in a subject population treated with the method is at least or about 15%. In another embodiment, the complete response rate in a subject population treated with the method is at least or about 20%.

[0361] Similarly, using the percentage of partial responses as a criterion, in one embodiment, a population of human subjects is treated by the methods provided herein, wherein the percentage of subjects in the treated population that have a partial response is at least or about 25%. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the percentage of subjects in the treated population that have a partial response is at least or about 30%. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the percentage of subjects in the treated population that have a partial response is at least or about 35%. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the percentage of subjects in the treated population that have a partial response is at least or about 40%. In one embodiment, a population of human subjects is treated by the methods provided herein, wherein the percentage of subjects in the treated population that have a partial response is at least or about 45%. In one embodiment, a population of human subjects is treated by the methods provided herein, wherein the percentage of subjects in the treated population that have a partial response is at least or about 50%. In one embodiment, a population of human subjects is treated by the methods provided herein, wherein the percentage of subjects in the treated population that have a partial response is at least or about 53.9%. In one embodiment, a human subject population is treated by the methods provided herein, wherein the percentage of subjects with partial responses in the treated population is at least or about 54%. In another embodiment, a human subject population is treated by the methods provided herein, wherein the percentage of subjects with partial responses in the treated population is at least or about 55%. In another embodiment, a human subject population is treated by the methods provided herein, wherein the percentage of subjects with partial responses in the treated population is at least or about 60%. In another embodiment, a human subject population is treated by the methods provided herein, wherein the percentage of subjects with partial responses in the treated population is at least or about 65%. In another embodiment, a human subject population is treated by the methods provided herein, wherein the percentage of subjects with partial responses in the treated population is at least or about 70%. In another embodiment, a human subject population is treated by the methods provided herein, wherein the percentage of subjects with partial responses in the treated population is at least or about 75%.

[0362] In one embodiment, the partial response rate in a subject population treated with the method is at least or about 25%. In another embodiment, the partial response rate in a subject population treated with the method is at least or about 30%. In another embodiment, the partial response rate in a subject population treated with the method is at least or about 35%. In another embodiment, the partial response rate in a subject population treated with the method is at least or about 40%. In another embodiment, the partial response rate in a subject population treated with the method is at least or about 45%. In another embodiment, the partial response rate in a subject population treated with the method is at least or about 50%. In another embodiment, the partial response rate in a subject population treated with the method is at least or about 53.9%. In another embodiment, the partial response rate in a subject population treated with the method is at least or about 54%. In another embodiment, the partial response rate in a subject population treated with the method is at least or about 55%. In another embodiment, the partial response rate in a subject population treated with the method is at least or about 60%. In another embodiment, the partial response rate in a subject population treated with the method is at least or about 65%. In another embodiment, the partial response rate in a subject population treated with the method is at least or about 70%. In another embodiment, the partial response rate in a population of subjects treated with the method is at least or about 75%.

[0363] In addition, the percentage of the subject of stable disease can be used as the evaluation standard of the treatment result of the method treatment human subject that this paper provides.In one embodiment, by the method treatment human subject colony that this paper provides, the percentage of the subject of stable disease in the wherein treated colony is at least or about 10%.In one embodiment, by the method treatment human subject colony that this paper provides, the percentage of the subject of stable disease in the wherein treated colony is at least or about 15%.In one embodiment, by the method treatment human subject colony that this paper provides, the percentage of the subject of stable disease in the wherein treated colony is at least or about 20%.In one embodiment, by the method treatment human subject colony that this paper provides, the percentage of the subject of stable disease in the wherein treated colony is at least or about 22%.In one embodiment, by the method treatment human subject colony that this paper provides, the percentage of the subject of stable disease in the wherein treated colony is at least or about 22.4%.In one embodiment, by the method treatment human subject colony that this paper provides, the percentage of the subject of stable disease in the wherein treated colony is at least or about 25%.In one embodiment, by the method treatment human subject colony that this paper provides, the percentage of the subject of stable disease in the wherein treated colony is at least or about 30%. In one embodiment, by the method treatment human subject colony that this paper provides, the percentage of the experimenter of wherein disease stable in the treated colony is at least or about 35%.In one embodiment, by the method treatment human subject colony that this paper provides, the percentage of the experimenter of wherein disease stable in the treated colony is at least or about 40%.In one embodiment, by the method treatment human subject colony that this paper provides, the percentage of the experimenter of wherein disease stable in the treated colony is at least or about 45%.In one embodiment, by the method treatment human subject colony that this paper provides, the percentage of the experimenter of wherein disease stable in the treated colony is at least or about 50%.

[0364] In one embodiment, the disease stabilization rate in a subject population treated with the method is at least or about 10%. In one embodiment, the disease stabilization rate in a subject population treated with the method is at least or about 15%. In one embodiment, the disease stabilization rate in a subject population treated with the method is at least or about 20%. In one embodiment, the disease stabilization rate in a subject population treated with the method is at least or about 22%. In one embodiment, the disease stabilization rate in a subject population treated with the method is at least or about 22.4%. In one embodiment, the disease stabilization rate in a subject population treated with the method is at least or about 25%. In one embodiment, the disease stabilization rate in a subject population treated with the method is at least or about 30%. In one embodiment, the disease stabilization rate in a subject population treated with the method is at least or about 35%. In one embodiment, the disease stabilization rate in a subject population treated with the method is at least or about 40%. In one embodiment, the disease stabilization rate in a subject population treated with the method is at least or about 45%. In one embodiment, the disease stabilization rate in a subject population treated with the method is at least or about 50%.

[0365] Similarly, the objective response rate, which is the sum of the percentage of subjects who have experienced a complete response and the percentage of subjects who have experienced a partial response, can be used as an evaluation criterion for the treatment results of human subjects treated by the methods provided herein. In another embodiment, a human subject population is treated by the methods provided herein, wherein the objective response rate of the treated population is at least or about 30%. In one embodiment, a human subject population is treated by the methods provided herein, wherein the objective response rate of the treated population is at least or about 35%. In another embodiment, a human subject population is treated by the methods provided herein, wherein the objective response rate of the treated population is at least or about 40%. In one embodiment, a human subject population is treated by the methods provided herein, wherein the objective response rate of the treated population is at least or about 45%. In another embodiment, a human subject population is treated by the methods provided herein, wherein the objective response rate of the treated population is at least or about 50%. In another embodiment, a human subject population is treated by the methods provided herein, wherein the objective response rate of the treated population is at least or about 52.7%. In another embodiment, a human subject population is treated by the methods provided herein, wherein the objective response rate of the treated population is at least or about 53%. In another embodiment, a human subject population is treated by the method provided herein, wherein the objective response rate of the treated population is at least or about 55%. In one embodiment, a human subject population is treated by the method provided herein, wherein the objective response rate of the treated population is at least or about 60%. In another embodiment, a human subject population is treated by the method provided herein, wherein the objective response rate of the treated population is at least or about 64.5%. In another embodiment, a human subject population is treated by the method provided herein, wherein the objective response rate of the treated population is at least or about 65%. In another embodiment, a human subject population is treated by the method provided herein, wherein the objective response rate of the treated population is at least or about 70%. In one embodiment, a human subject population is treated by the method provided herein, wherein the objective response rate of the treated population is at least or about 75%. In one embodiment, a human subject population is treated by the method provided herein, wherein the objective response rate of the treated population is at least or about 75.1%. In another embodiment, a human subject population is treated by the method provided herein, wherein the objective response rate of the treated population is at least or about 80%. In another embodiment, a human subject population is treated by the method provided herein, wherein the objective response rate of the treated population is at least or about 85%. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the objective response rate in the treated population is at least or about 90%.

[0366] In one embodiment, a population of human subjects is treated by the methods provided herein, wherein the objective response rate of the treated population is in the range of 40% to 80%. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the objective response rate of the treated population is in the range of 40% to 75%. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the objective response rate of the treated population is in the range of 40% to 70%. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the objective response rate of the treated population is in the range of 45% to 80%. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the objective response rate of the treated population is in the range of 45% to 75%. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the objective response rate of the treated population is in the range of 45% to 70%. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the objective response rate of the treated population is in the range of 50% to 80%. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the objective response rate of the treated population is in the range of 50% to 75%. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the objective response rate of the treated population is in the range of 52.7% to 75.1%. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the objective response rate of the treated population is in the range of 55% to 80%. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the objective response rate of the treated population is in the range of 55% to 75%. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the objective response rate of the treated population is in the range of 55% to 70%. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the objective response rate of the treated population is in the range of 55% to 65%. In one embodiment, a population of human subjects is treated by the methods provided herein, wherein the objective response rate of the treated population is in the range of 60% to 80%. In one embodiment, a human subject population is treated by the methods provided herein, wherein the objective response rate of the treated population is in the range of 60% to 75%. In one embodiment, a human subject population is treated by the methods provided herein, wherein the objective response rate of the treated population is in the range of 60% to 70%. In one embodiment, a human subject population is treated by the methods provided herein, wherein the objective response rate of the treated population is in the range of 60% to 65%.

[0367] In one embodiment, the objective response rate of the subject population treated with the method is at least or about 30%. In another embodiment, the objective response rate of the subject population treated with the method is at least or about 30%. In another embodiment, the objective response rate of the subject population treated with the method is at least or about 35%. In another embodiment, the objective response rate of the subject population treated with the method is at least or about 40%. In another embodiment, the objective response rate of the subject population treated with the method is at least or about 45%. In another embodiment, the objective response rate of the subject population treated with the method is at least or about 50%. In another embodiment, the objective response rate of the subject population treated with the method is at least or about 55%. In another embodiment, the objective response rate of the subject population treated with the method is at least or about 60%. In another embodiment, the objective response rate of the subject population treated with the method is at least or about 64.5%. In another embodiment, the objective response rate of the subject population treated with the method is at least or about 65%. In another embodiment, the objective response rate of the subject population treated with the method is at least or about 70%. In another embodiment, the objective response rate in a subject population treated with the method is at least or about 75%. In another embodiment, the objective response rate in a subject population treated with the method is at least or about 80%. In another embodiment, the objective response rate in a subject population treated with the method is at least or about 85%. In another embodiment, the objective response rate in a subject population treated with the method is at least or about 90%.

[0368] In one embodiment, the objective response rate in a subject population treated with the method is 40% to 80%. In another embodiment, the objective response rate in a subject population treated with the method is 40% to 75%. In another embodiment, the objective response rate in a subject population treated with the method is 40% to 70%. In another embodiment, the objective response rate in a subject population treated with the method is 45% to 80%. In another embodiment, the objective response rate in a subject population treated with the method is 45% to 75%. In another embodiment, the objective response rate in a subject population treated with the method is 45% to 70%. In another embodiment, the objective response rate in a subject population treated with the method is 50% to 80%. In another embodiment, the objective response rate in a subject population treated with the method is 50% to 75%. In another embodiment, the objective response rate in a subject population treated with the method is 52.7% to 75.1%. In another embodiment, the objective response rate in a subject population treated with the method is 53% to 75%. In another embodiment, the objective response rate in a subject population treated with the method is 55% to 80%. In another embodiment, the objective response rate in the subject population treated with the method is 55% to 75%. In another embodiment, the objective response rate in the subject population treated with the method is 55% to 70%. In another embodiment, the objective response rate in the subject population treated with the method is 55% to 65%. In another embodiment, the objective response rate in the subject population treated with the method is 60% to 80%. In another embodiment, the objective response rate in the subject population treated with the method is 60% to 75%. In another embodiment, the objective response rate in the subject population treated with the method is 60% to 70%. In another embodiment, the objective response rate in the subject population treated with the method is 60% to 65%.

[0369] In addition, the treatment outcome of the methods provided herein can be evaluated based on the duration of response as described in Section 6.1.8.2(ii). In one embodiment, the duration of response of the human subject after treatment is at least or about 5 months. In another embodiment, the duration of response of the human subject after treatment is at least or about 6 months. In another embodiment, the duration of response of the human subject after treatment is at least or about 7 months. In another embodiment, the duration of response of the human subject after treatment is at least or about 8 months. In one embodiment, the duration of response of the human subject after treatment is at least or about 9 months. In another embodiment, the duration of response of the human subject after treatment is at least or about 10 months. In another embodiment, the duration of response of the human subject after treatment is at least or about 11 months. In one embodiment, the duration of response of the human subject after treatment is at least or about 12 months. In another embodiment, the duration of response of the human subject after treatment is at least or about 13 months. In another embodiment, the duration of response of the human subject after treatment is at least or about 14 months. In another embodiment, the duration of response of the human subject after treatment is at least or about 15 months. In one embodiment, the duration of response in a human subject after treatment is at least or about 16 months. In another embodiment, the duration of response in a human subject after treatment is at least or about 17 months. In another embodiment, the duration of response in a human subject after treatment is at least or about 18 months. In another embodiment, the duration of response in a human subject after treatment is at least or about 19 months. In another embodiment, the duration of response in a human subject after treatment is at least or about 20 months. In another embodiment, the duration of response in a human subject after treatment is at least or about 21 months. In another embodiment, the duration of response in a human subject after treatment is at least or about 22 months. In another embodiment, the duration of response in a human subject after treatment is at least or about 23 months. In another embodiment, the duration of response in a human subject after treatment is at least or about 24 months. In another embodiment, the duration of response in a human subject after treatment is at least or about 25 months. In another embodiment, the duration of response in a human subject after treatment is at least or about 26 months. In another embodiment, the duration of response in a human subject after treatment is at least or about 27 months. In another embodiment, the duration of response in a human subject after treatment is at least or about 28 months. In another embodiment, the duration of response in a human subject following treatment is at least or about 29 months. In another embodiment, the duration of response in a human subject following treatment is at least or about 30 months.

[0370] In another embodiment, the duration of response in a human subject after treatment ranges from 5 to 30 months. In another embodiment, the duration of response in a human subject after treatment ranges from 5 to 29 months. In another embodiment, the duration of response in a human subject after treatment ranges from 5 to 28 months. In another embodiment, the duration of response in a human subject after treatment ranges from 5 to 27 months. In another embodiment, the duration of response in a human subject after treatment ranges from 5 to 26 months. In another embodiment, the duration of response in a human subject after treatment ranges from 5 to 25 months. In another embodiment, the duration of response in a human subject after treatment ranges from 5 to 24 months. In another embodiment, the duration of response in a human subject after treatment ranges from 5 to 23 months. In some embodiments, the duration of response in a human subject after treatment ranges from 5 to 22 months. In another embodiment, the duration of response in a human subject after treatment ranges from 5 to 21 months. In another embodiment, the duration of response in a human subject after treatment ranges from 5 to 20 months. In another embodiment, the duration of response in a human subject after treatment ranges from 5 to 19 months. In one embodiment, the duration of response in a human subject after treatment ranges from 5 to 18 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 5 to 17 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 5 to 16 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 5 to 15 months. In one embodiment, the duration of response in a human subject after treatment is in the range of 5 to 14 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 5 to 13 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 5 to 12 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 6 to 30 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 6 to 29 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 6 to 28 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 6 to 27 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 6 to 26 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 6 to 25 months. In another embodiment, the duration of response in a human subject following treatment ranges from 6 to 24 months.In another embodiment, the duration of response in a human subject after treatment is in the range of 6 to 23 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 6 to 22 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 6 to 21 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 6 to 20 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 6 to 19 months. In one embodiment, the duration of response in a human subject after treatment is in the range of 6 to 18 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 6 to 17 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 6 to 16 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 6 to 15 months. In one embodiment, the duration of response in a human subject after treatment is in the range of 6 to 14 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 6 to 13 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 6 to 12 months. In one embodiment, the duration of response in a human subject after treatment is in the range of 7 to 22 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 7 to 21 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 7 to 20 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 7 to 19 months. In one embodiment, the duration of response in a human subject after treatment is in the range of 7 to 18 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 7 to 17 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 7 to 16 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 7 to 15 months. In one embodiment, the duration of response in a human subject after treatment is in the range of 7 to 14 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 7 to 13 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 7 to 12 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 8 to 30 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 9 to 30 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 10 to 30 months.In another embodiment, the duration of response of a human subject after treatment ranges from 11 to 30 months. In one embodiment, the duration of response of a human subject after treatment ranges from 12 to 30 months. In another embodiment, the duration of response of a human subject after treatment ranges from 13 to 30 months. In another embodiment, the duration of response of a human subject after treatment ranges from 14 to 30 months. In another embodiment, the duration of response of a human subject after treatment ranges from 15 to 30 months. In another embodiment, the duration of response of a human subject after treatment ranges from 16 to 30 months. In one embodiment, the duration of response of a human subject after treatment ranges from 17 to 30 months. In another embodiment, the duration of response of a human subject after treatment ranges from 18 to 30 months. In another embodiment, the duration of response of a human subject after treatment ranges from 8 to 27 months. In another embodiment, the duration of response of a human subject after treatment ranges from 9 to 27 months. In another embodiment, the duration of response of a human subject after treatment ranges from 10 to 27 months. In another embodiment, the duration of response of a human subject after treatment ranges from 11 to 27 months. In one embodiment, the duration of response in a human subject after treatment is in the range of 12 to 27 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 13 to 27 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 14 to 27 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 15 to 27 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 16 to 27 months. In one embodiment, the duration of response in a human subject after treatment is in the range of 17 to 27 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 18 to 27 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 8 to 22 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 9 to 22 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 10 to 22 months. In another embodiment, the duration of response in a human subject after treatment is in the range of 11 to 22 months. In one embodiment, the duration of response in a human subject after treatment is in the range of 12 to 22 months. In another embodiment, the duration of response in human subjects following treatment ranges from 13 to 22 months.In another embodiment, the duration of response in a human subject after treatment ranges from 14 to 22 months. In another embodiment, the duration of response in a human subject after treatment ranges from 15 to 22 months. In another embodiment, the duration of response in a human subject after treatment ranges from 16 to 22 months. In one embodiment, the duration of response in a human subject after treatment ranges from 17 to 22 months. In another embodiment, the duration of response in a human subject after treatment ranges from 18 to 22 months. In another embodiment, the duration of response in a human subject after treatment ranges from 6 to 21 months. In another embodiment, the duration of response in a human subject after treatment ranges from 7 to 20 months. In another embodiment, the duration of response in a human subject after treatment ranges from 8 to 19 months. In one embodiment, the duration of response in a human subject after treatment ranges from 9 to 18 months. In another embodiment, the duration of response in a human subject after treatment ranges from 10 to 17 months. In another embodiment, the duration of response in a human subject after treatment ranges from 11 to 16 months. In another embodiment, the duration of response in a human subject after treatment ranges from 12 to 15 months. In another embodiment, the duration of response in human subjects following treatment ranges from 13 to 14 months.

[0371] In certain embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response of the treated population after treatment is at least or about 5 months. In certain embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response of the treated population after treatment is at least or about 6 months. In certain embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response of the treated population after treatment is at least or about 7 months. In certain embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response of the treated population after treatment is at least or about 8 months. In certain embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response of the treated population after treatment is at least or about 9 months. In certain embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response of the treated population after treatment is at least or about 10 months. In certain embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response of the treated population after treatment is at least or about 11 months. In certain embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response of the treated population after treatment is at least or about 12 months. In certain embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response of the treated population after treatment is at least or about 13 months. In certain embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response of the treated population after treatment is at least or about 14 months. In certain embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response of the treated population after treatment is at least or about 15 months. In certain embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response of the treated population after treatment is at least or about 16 months. In certain embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response of the treated population after treatment is at least or about 17 months. In certain embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response of the treated population after treatment is at least or about 18 months. In certain embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response of the treated population after treatment is at least or about 19 months. In certain embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response of the treated population after treatment is at least or about 20 months. In certain embodiments, a population of human subjects is treated by the methods provided herein, wherein the duration of response in the treated population following treatment is at least or about 21 months.In certain embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response of the treated population after treatment is at least or about 22 months. In certain embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response of the treated population after treatment is at least or about 23 months. In certain embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response of the treated population after treatment is at least or about 24 months. In certain embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response of the treated population after treatment is at least or about 25 months. In certain embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response of the treated population after treatment is at least or about 26 months. In certain embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response of the treated population after treatment is at least or about 27 months. In certain embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response of the treated population after treatment is at least or about 28 months. In certain embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response of the treated population after treatment is at least or about 29 months. In certain embodiments, a population of human subjects is treated by the methods provided herein, wherein the duration of response in the treated population following treatment is at least or about 30 months.

[0372] In certain embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 5 to 30 months. In some embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 5 to 29 months. In some embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 5 to 28 months. In some embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 5 to 27 months. In some embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 5 to 26 months. In some embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 5 to 25 months. In some embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 5 to 24 months. In some embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 5 to 23 months. In certain embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 5 to 22 months. In some embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 5 to 21 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 5 to 20 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 5 to 19 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 5 to 18 months. In one embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 5 to 17 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 5 to 16 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 5 to 15 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 5 to 14 months. In one embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 5 to 13 months.In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 5 to 12 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 6 to 30 months. In some embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 6 to 29 months. In some embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 6 to 28 months. In some embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 6 to 27 months. In some embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 6 to 26 months. In some embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 6 to 25 months. In some embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 6 to 24 months. In some embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 6 to 23 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 6 to 22 months. In some embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 6 to 21 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 6 to 20 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 6 to 19 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 6 to 18 months. In one embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 6 to 17 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 6 to 16 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 6 to 15 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 6 to 14 months.In one embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 6 to 13 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 6 to 12 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 7 to 30 months. In some embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 7 to 29 months. In some embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 7 to 28 months. In some embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 7 to 27 months. In some embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 7 to 26 months. In some embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 7 to 25 months. In some embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 7 to 24 months. In some embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 7 to 23 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 7 to 22 months. In some embodiments, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 7 to 21 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 7 to 20 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 7 to 19 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 7 to 18 months. In one embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 7 to 17 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 7 to 16 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 7 to 15 months.In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 7 to 14 months. In one embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 7 to 13 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 7 to 12 months. In one embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 6.41 to 22 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 8 to 22 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 9 to 22 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 10 to 22 months. In one embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 11 to 22 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is within the range of 12 to 12 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is within the range of 13 to 22 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is within the range of 14 to 22 months. In one embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is within the range of 15 to 22 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is within the range of 16 to 22 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is within the range of 17 to 22 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is within the range of 18 to 22 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is within the range of 7 to 27 months. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the duration of response in the treated population ranges from 8 to 27 months.In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 9 to 27 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 10 to 27 months. In one embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 11 to 27 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 12 to 27 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 13 to 27 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 14 to 27 months. In one embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 15 to 27 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 16 to 27 months. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 17 to 27 months. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 18 to 27 months. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 19 to 27 months. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 20 to 27 months. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 21 to 27 months. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 22 to 27 months. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 23 to 27 months. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 24 to 27 months. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the duration of response in the treated population ranges from 7 to 30 months.In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 8 to 30 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 9 to 30 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 10 to 30 months. In one embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 11 to 30 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 12 to 30 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 13 to 30 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 14 to 30 months. In one embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 15 to 30 months. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 16 to 30 months. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 17 to 30 months. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 18 to 30 months. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 19 to 30 months. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 20 to 30 months. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 21 to 30 months. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 22 to 30 months. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 23 to 30 months. In another embodiment, a population of human subjects is treated by the methods provided herein, wherein the duration of response in the treated population ranges from 24 to 30 months.In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 25 to 30 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 26 to 30 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 27 to 30 months. In one embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 6 to 21 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 7 to 20 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 8 to 19 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 9 to 18 months. In one embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 10 to 17 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 11 to 16 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 12 to 15 months. In another embodiment, a human subject population is treated by the methods provided herein, wherein the duration of response in the treated population is in the range of 13 to 24 months.

[0373] In certain embodiments, the duration of response in a subject population treated with the method is at least or about 5 months. In certain embodiments, the duration of response in a subject population treated with the method is at least or about 6 months. In certain embodiments, the duration of response in a subject population treated with the method is at least or about 7 months. In certain embodiments, the duration of response in a subject population treated with the method is at least or about 8 months. In certain embodiments, the duration of response in a subject population treated with the method is at least or about 9 months. In certain embodiments, the duration of response in a subject population treated with the method is at least or about 10 months. In certain embodiments, the duration of response in a subject population treated with the method is at least or about 11 months. In certain embodiments, the duration of response in a subject population treated with the method is at least or about 12 months. In certain embodiments, the duration of response in a subject population treated with the method is at least or about 13 months. In certain embodiments, the duration of response in a subject population treated with the method is at least or about 14 months. In certain embodiments, the duration of response in a subject population treated with the method is at least or about 15 months. In certain embodiments, the duration of response in a subject population treated with the method is at least or about 16 months. In certain embodiments, the duration of response in a subject population treated with the method is at least or about 17 months. In certain embodiments, the duration of response in a subject population treated with the method is at least or about 18 months. In certain embodiments, the duration of response in a subject population treated with the method is at least or about 19 months. In certain embodiments, the duration of response in a subject population treated with the method is at least or about 20 months. In certain embodiments, the duration of response in a subject population treated with the method is at least or about 21 months. In certain embodiments, the duration of response in a subject population treated with the method is at least or about 22 months. In certain embodiments, the duration of response in a subject population treated with the method is at least or about 23 months. In certain embodiments, the duration of response in a subject population treated with the method is at least or about 24 months. In certain embodiments, the duration of response in a subject population treated with the method is at least or about 25 months. In certain embodiments, the duration of response in a subject population treated with the method is at least or about 26 months. In certain embodiments, the duration of response in a subject population treated with the methods is at least or about 27 months. In certain embodiments, the duration of response in a subject population treated with the methods is at least or about 28 months. In certain embodiments, the duration of response in a subject population treated with the methods is at least or about 29 months.In certain embodiments, the duration of response in a population of subjects treated with the methods is at least or about 30 months.

[0374] Alternatively, the treatment outcome of the methods provided herein can be evaluated based on progression-free survival as described in Section 6.1.8.2 (iv). In one embodiment, the human subject has a progression-free survival of at least or about 5 months after treatment. In another embodiment, the human subject has a progression-free survival of at least or about 6 months after treatment. In another embodiment, the human subject has a progression-free survival of at least or about 7 months after treatment. In another embodiment, the human subject has a progression-free survival of at least or about 8 months after treatment. In one embodiment, the human subject has a progression-free survival of at least or about 9 months after treatment. In another embodiment, the human subject has a progression-free survival of at least or about 10 months after treatment. In another embodiment, the human subject has a progression-free survival of at least or about 11 months after treatment. In another embodiment, the human subject has a progression-free survival of at least or about 12 months after treatment. In one embodiment, the human subject has a progression-free survival of at least or about 13 months after treatment. In another embodiment, the human subject has a progression-free survival of at least or about 14 months after treatment. In another embodiment, the human subject has a progression-free survival period of at least or about 15 months after treatment. In another embodiment, the human subject has a progression-free survival period of at least or about 16 months after treatment. In one embodiment, the human subject has a progression-free survival period of at least or about 17 months after treatment. In another embodiment, the human subject has a progression-free survival period of at least or about 18 months after treatment. In another embodiment, the human subject has a progression-free survival period of at least or about 19 months after treatment. In another embodiment, the human subject has a progression-free survival period of at least or about 20 months after treatment. In another embodiment, the human subject has a progression-free survival period of at least or about 21 months after treatment. In another embodiment, the human subject has a progression-free survival period of at least or about 22 months after treatment. In another embodiment, the human subject has a progression-free survival period of at least or about 23 months after treatment. In another embodiment, the human subject has a progression-free survival period of at least or about 24 months after treatment. In another embodiment, the human subject has a progression-free survival period of at least or about 25 months after treatment. In another embodiment, the human subject has a progression-free survival of at least or about 26 months following treatment. In another embodiment, the human subject has a progression-free survival of at least or about 27 months following treatment. In another embodiment, the human subject has a progression-free survival of at least or about 28 months following treatment. In another embodiment, the human subject has a progression-free survival of at least or about 29 months following treatment.In another embodiment, the human subject has a progression-free survival of at least or about 30 months following treatment. In another embodiment, the human subject has a progression-free survival of at least or about 31 months following treatment. In another embodiment, the human subject has a progression-free survival of at least or about 32 months following treatment. In another embodiment, the human subject has a progression-free survival of at least or about 33 months following treatment.

[0375] In another embodiment, the human subject has a progression-free survival period in the range of 5 to 33 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 5 to 32 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 5 to 31 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 5 to 30 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 5 to 29 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 5 to 28 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 5 to 27 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 5 to 26 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 5 to 25 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 5 to 24 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 5 to 23 months after treatment. In some embodiments, the human subject has a progression-free survival period in the range of 5 to 22 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 5 to 21 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 5 to 20 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 5 to 19 months after treatment. In one embodiment, the human subject has a progression-free survival period in the range of 5 to 18 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 5 to 17 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 5 to 16 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 5 to 15 months after treatment. In one embodiment, the human subject has a progression-free survival period in the range of 5 to 14 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 5 to 13 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 5 to 12 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 6 to 33 months following treatment. In another embodiment, the human subject has a progression-free survival period in the range of 6 to 32 months following treatment. In another embodiment, the human subject has a progression-free survival period in the range of 6 to 31 months following treatment.In another embodiment, the human subject has a progression-free survival period in the range of 6 to 30 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 6 to 29 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 6 to 28 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 6 to 27 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 6 to 26 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 6 to 25 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 6 to 22 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 6 to 24 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 6 to 23 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 6 to 22 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 6 to 21 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 6 to 20 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 6 to 19 months after treatment. In one embodiment, the human subject has a progression-free survival period in the range of 6 to 18 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 6 to 17 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 6 to 16 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 6 to 15 months after treatment. In one embodiment, the human subject has a progression-free survival period in the range of 6 to 14 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 6 to 13 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 6 to 12 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 7 to 33 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 7 to 32 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 7 to 31 months following treatment. In another embodiment, the human subject has a progression-free survival period in the range of 7 to 30 months following treatment. In another embodiment, the human subject has a progression-free survival period in the range of 7 to 29 months following treatment.In another embodiment, the human subject has a progression-free survival period in the range of 7 to 28 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 7 to 27 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 7 to 26 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 7 to 25 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 7 to 24 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 7 to 23 months after treatment. In one embodiment, the human subject has a progression-free survival period in the range of 7 to 22 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 7 to 21 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 7 to 20 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 7 to 19 months after treatment. In one embodiment, the human subject has a progression-free survival period in the range of 7 to 18 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 7 to 17 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 7 to 16 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 7 to 15 months after treatment. In one embodiment, the human subject has a progression-free survival period in the range of 7 to 14 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 7 to 13 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 7 to 12 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 8 to 22 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 9 to 22 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 10 to 22 months after treatment. In another embodiment, the human subject has a progression-free survival period in the range of 11 to 22 month...

Claims

1. A method of treating cancer in a human subject, the method comprising administering to the subject: (a) an effective amount of an antibody-drug conjugate (ADC) comprising an anti-191P4D12 antibody or an antigen-binding fragment thereof and (b) an effective amount of an anti-PD-1 antibody; wherein the anti-191P4D12 antibody or antigen-binding fragment thereof binds to 191P4D12 and is coupled to one or more units of monomethyl auristatin E (MMAE); wherein the anti-PD-1 antibody comprises: (i) a light chain variable region comprising light chain CDR1, CDR2 and CDR3 of SEQ ID NOs: 24, 25 and 26, respectively, and (ii) a heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 of SEQ ID NOs: 29, 30 and 31, respectively; wherein the anti-191P4D12 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 shown 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 shown in SEQ ID NO: 23; wherein the subject suffers from urothelial carcinoma or bladder cancer; wherein the subject has not received immune checkpoint inhibitor (CPI) therapy; and wherein the subject is not suitable for treatment with cisplatin (cisplatin ineligible).

2. The method of claim 1, wherein the subject has visceral metastases.

3. The method of claim 1, wherein the subject has lymph node-only disease.

4. The method of any one of claims 1 to 3, wherein the site of origin of the disease is in the upper urinary tract.

5. The method of any one of claims 1 to 3, wherein the site of origin of the disease is in the lower urinary tract.

6. The method of any one of claims 1 to 5, wherein the subject has a combined positive score (CPS) for PD-L1 expression greater than or equal to 10.

7. The method of any one of claims 1 to 5, wherein the subject has a PD-L1 expression CPS of less than 10.

8. The method of any one of claims 1 to 6, wherein the subject has a Connexin-4H-Fraction between 0 and 300.

9. The method of any one of claims 1 to 7, wherein the subject has a Connexin-4H-Fraction between 0 and 200.

10. The method of any one of claims 1 to 9, wherein the subject has an ECOG performance status of 1 to 2.

11. The method of any one of claims 1 to 9, wherein the subject has one or more conditions selected from the group consisting of: an ECOG performance status of 2, impaired renal function, and hearing loss of not less than grade 2.

12. The method of any one of claims 1 to 9, wherein the subject has NYHA class III heart failure.

13. The method of claim 11, wherein the subject has an ECOG performance status of 2, and The subject has (i) hemoglobin ≥ 10 g / dL; (ii) GFR ≥ 50 mL / min; and (iii) no NYHA class III heart failure.

14. The method of claim 11, wherein the impaired renal function is determined based on a creatinine clearance (CrCl) of less than 60 mL / min.

15. The method of claim 11, wherein the impaired renal function is determined based on a CrCl of less than 60 but not less than 30 mL / min.

16. The method of claim 11, wherein the impaired renal function is determined based on a CrCl of less than 30 but not less than 15 mL / min.

17. The method of any one of claims 1 to 16, wherein the subject has one or more conditions selected from the group consisting of: (i) absolute neutrophil count not less than 1500 / μL; (ii) platelet count not less than 100,000 / μL; (iii) hemoglobin not less than 9 g / dL; (iv) serum bilirubin not exceeding 1.5 times the upper limit of normal (ULN) or not exceeding 3 times the ULN for patients with Gilbert's disease; (v) CrCl not less than 30 mL / min, and (vi) Alanine aminotransferase and aspartate aminotransferase not exceeding 3 times the ULN.

18. The method of claim 17, wherein the subject has all of conditions (i) to (vi) of claim 15.

19. The method of any one of claims 14 to 18, wherein the CrCl is measured by 24-hour urine collection or estimated according to the Croft-Gault criteria.

20. The method of any one of claims 1 to 19, wherein the subject does not suffer from sensory or motor neuropathy greater than grade 2.

21. The method of any one of claims 1 to 20, wherein the subject does not have active central nervous system metastases.

22. The method of any one of claims 1 to 21, wherein the subject does not have uncontrolled diabetes.

23. The method of claim 22, wherein the uncontrolled diabetes is determined by a hemoglobin A1c (HbA1c) not less than 8% or a HbA1c between 7% and 8% accompanied by associated diabetic symptoms not otherwise specified.

24. The method of claim 23, wherein the associated diabetes symptoms comprise or consist of polyuria, polydipsia, or both polyuria and polydipsia.

25. The method of any one of claims 1 to 24, wherein the subject has locally advanced or metastatic urothelial carcinoma.

26. The method of any one of claims 1 to 25, wherein the subject has locally advanced or metastatic bladder cancer.

27. The method of any one of claims 1 to 26, wherein the anti-191P4D12 antibody or antigen-binding fragment thereof comprises: a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 9, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 10, a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 11; a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 12, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 13, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 14, or The anti-191P4D12 antibody or its antigen-binding fragment 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, and 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.

28. The method of any one of claims 1 to 26, wherein the anti-191P4D12 antibody or antigen-binding fragment thereof comprises: a CDR-H1 consisting of the amino acid sequence of SEQ ID NO: 9, a CDR-H2 consisting of the amino acid sequence of SEQ ID NO: 10, a CDR-H3 consisting of the amino acid sequence of SEQ ID NO: 11; a CDR-L1 consisting of the amino acid sequence of SEQ ID NO: 12, a CDR-L2 consisting of the amino acid sequence of SEQ ID NO: 13, and a CDR-L3 consisting of the amino acid sequence of SEQ ID NO: 14, or The anti-191P4D12 antibody or its antigen-binding fragment comprises: CDR-H1 composed of the amino acid sequence SEQ ID NO: 16, CDR-H2 composed of the amino acid sequence SEQ ID NO: 17, CDR-H3 composed of the amino acid sequence SEQ ID NO: 18; CDR-L1 composed of the amino acid sequence SEQ ID NO: 19, CDR-L2 composed of the amino acid sequence SEQ ID NO: 20 and CDR-L3 composed of the amino acid sequence SEQ ID NO:

21.

29. The method of any one of claims 1 to 28, wherein the anti-191P4D12 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.

30. The method of any one of claims 1 to 29, wherein the anti-191P4D12 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.

31. The method of any one of claims 1 to 30, wherein the anti-191P4D12 antigen-binding fragment is Fab, F(abt)2, Fv or scFv.

32. The method of any one of claims 1 to 30, wherein the anti-191P4D12 antibody is a fully human antibody.

33. The method of any one of claims 1 to 30 and 32, wherein the anti-191P4D12 antibody is IgG1 and the light chain is a kappa light chain.

34. The method of any one of claims 1 to 33, wherein the anti-191P4D12 antibody or antigen-binding fragment thereof is recombinantly produced.

35. The method of any one of claims 1 to 34, wherein the anti-191P4D12 antibody or antigen-binding fragment is coupled to each unit of MMAE via a linker.

36. The method of claim 35, wherein the linker is an enzyme-cleavable linker, and wherein the linker forms a bond with a sulfur atom of the antibody or antigen-binding fragment thereof.

37. A method as claimed in claim 35 or 36, wherein the linker has the formula: -Aa-Ww-Yy-; wherein -A- is a stretcher unit, a is 0 or 1; -W- is an amino acid unit, w is an integer in the range of 0 to 12; and -Y- is a spacer unit, y is 0, 1 or 2.

38. The method of claim 37, wherein the stretcher unit has the structure of formula (1); the amino acid unit is valine-citrulline; and the spacer unit is a PAB group comprising the structure of formula (2):

39. The method of claim 37 or 38, wherein the Stretcher unit forms a bond with a sulfur atom of the antibody or antigen-binding fragment thereof; and wherein the Spacer unit is linked to MMAE via a carbamate group.

40. The method of any one of claims 1 to 39, wherein the ADC comprises 1 to 20 MMAE units per antibody or antigen-binding fragment thereof.

41. The method of any one of claims 1 to 40, wherein the ADC comprises 1 to 10 MMAE units per antibody or antigen-binding fragment thereof.

42. The method of any one of claims 1 to 41, wherein the ADC comprises 2 to 8 MMAE units per antibody or antigen-binding fragment thereof.

43. The method of any one of claims 1 to 42, wherein the ADC comprises 3 to 5 MMAE units per antibody or antigen-binding fragment thereof.

44. The method of any one of claims 1 to 43, wherein the ADC has the following structure: wherein L- represents the anti-191P4D12 antibody or an antigen-binding fragment thereof and p is 1 to 10.

45. The method of claim 44, wherein p is 2 to 8.

46. ​​The method of claim 44 or 45, wherein p is 3 to 5.

47. The method of any one of claims 44 to 46, wherein p is 3 to 4.

48. The method of any one of claims 44 to 47, wherein p is about 4.

49. The method of any one of claims 44 to 47, wherein the effective amount of the antibody drug conjugate has a mean p-value of about 3.

8.

50. The method of any one of claims 1 to 49, wherein the ADC is administered to the subject at a dose of 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.

51. The method of any one of claims 1 to 50, wherein the ADC is administered to the subject at a dose of 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.

52. The method of any one of claims 1 to 51, wherein the ADC is administered to the subject at a dose of about 1 mg / kg of the subject's body weight.

53. The method of any one of claims 1 to 51, wherein the ADC is administered to the subject at a dose of about 1.25 mg / kg of the subject's body weight.

54. The method of any one of claims 1 to 53, wherein the ADC is administered to the subject by intravenous (IV) injection or infusion.

55. The method of any one of claims 1 to 54, wherein the ADC is administered to the subject by IV injection or infusion on up to 2 days within a 21 day treatment cycle.

56. The method of any one of claims 1 to 55, wherein the ADC is administered to the subject by IV injection or infusion on Days 1 and 8 of a 21 -day treatment cycle.

57. The method of any one of claims 1 to 56, wherein the ADC is administered to the subject by IV injection or infusion over about 30 minutes on up to 2 days within a 21 day treatment cycle.

58. The method of any one of claims 1 to 57, wherein the ADC is administered by IV injection or infusion over about 30 minutes on Days 1 and 8 of a 21 -day treatment cycle.

59. The method of any one of claims 1 to 58, wherein the ADC is formulated as a pharmaceutical composition comprising L-histidine, polysorbate-20 (TWEEN-20), and trehalose dihydrate.

60. The method of any one of claims 1 to 59, wherein the ADC is formulated as 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 wherein the pharmaceutical composition has a pH of about 6.0 at 25°C.

61. The method of any one of claims 1 to 59, wherein the ADC is formulated as 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 pharmaceutical composition has a pH of about 6.0 at 25°C.

62. The method of any one of claims 1 to 61, wherein the ADC has the following structure: wherein L- represents the antibody or antigen-binding fragment thereof and p is from about 3 to about 4, the anti-191P4D12 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, wherein the ADC is administered at a dose of about 1.25 mg / kg of the subject's body weight, and wherein the dose is administered by IV injection or infusion over about 30 minutes on days 1 and 8 of a 21-day treatment cycle.

63. The method of any one of claims 1 to 60, wherein the anti-PD-1 antibody is administered to the subject at a dose of about 100 mg to about 400 mg.

64. The method of any one of claims 1 to 61, wherein the anti-PD-1 antibody is administered to the subject at a dose of about 200 mg.

65. The method of claim 62, wherein: (a) the anti-PD-1 antibody is administered to the subject at a dose of about 200 mg; as well as (b) after step (a), the anti-PD-1 antibody is administered to the subject at a dose of about 400 mg every 42 days.

66. The method of any one of claims 1 to 65, wherein the anti-PD-1 antibody is administered to the subject by IV infusion on day 1 of the 21-day treatment cycle.

67. The method of any one of claims 1 to 66, wherein the anti-PD-1 antibody is administered to the subject by IV infusion on Day 1 of each 21 -day treatment cycle.

68. The method of any one of claims 1 to 67, wherein the anti-PD-1 antibody is administered to the subject by IV infusion over about 30 minutes on day 1 of the 21 -day treatment cycle.

69. The method of any one of claims 1 to 66, wherein the anti-PD-1 antibody is administered to the subject by IV infusion over about 30 minutes on Day 1 of each 21 -day treatment cycle.

70. The method of any one of claims 1 to 69, wherein the subject has a complete response following the treatment.

71. The method of any one of claims 1 to 69, wherein the subject has a partial response following the treatment.

72. The method of any one of claims 1 to 69, wherein the subject has a complete response or a partial response following the treatment.

73. The method of any one of claims 1 to 69, wherein the subject's disease is stabilized following the treatment.

74. The method of any one of claims 1 to 69, wherein the subject has a duration of response of at least or about 6 months following the treatment.

75. The method of any one of claims 1 to 69, wherein the subject has a duration of response of at least or about 12 months following the treatment.

76. The method of any one of claims 1 to 69, wherein the subject has a duration of response of at least or about 24 months following the treatment.

77. The method of any one of claims 1 to 69, wherein the subject has a duration of response of at least or about 27 months following the treatment.

78. The method of any one of claims 1 to 69, wherein the subject has a progression-free survival of at least or about 6 months following the treatment.

79. The method of any one of claims 1 to 69, wherein the subject has a progression-free survival of at least or about 8 months following the treatment.

80. The method of any one of claims 1 to 69, wherein the subject has a progression-free survival of at least or about 12 months following the treatment.

81. The method of any one of claims 1 to 69, wherein the subject has a progression-free survival of at least or about 20 months following the treatment.

82. The method of any one of claims 1 to 69, wherein the subject has a progression-free survival of at least or about 29 months following the treatment.

83. The method of any one of claims 1 to 69, wherein the subject has an overall survival of at least or about 22 months following the treatment.

84. The method of any one of claims 1 to 69, wherein the subject has an overall survival of at least or about 27 months following the treatment.

85. The method of any one of claims 1 to 69, wherein the subject has an overall survival of at least or about 30 months after treatment.

86. The method of any one of claims 1 to 69, wherein the subject has an overall survival in the range of 19 to 25 months following treatment.

87. The method of any one of claims 1 to 69, wherein the subject has an overall survival in the range of 28 to 32 months following treatment.

88. The method of any one of claims 1 to 69, wherein a population of subjects is treated by the method and wherein the percentage of subjects with a complete response in the treated population is at least or about 10%.

89. The method of any one of claims 1 to 69, wherein a population of subjects is treated by the method and wherein the percentage of subjects with a partial response in the treated population is at least or about 54%.

90. The method of any one of claims 1 to 69, wherein a population of the subjects is treated by the method, and wherein the objective response rate in the treated population is at least or about 65%.

91. The method of any one of claims 1 to 69, wherein a population of said subjects is treated by said method, and wherein the objective response rate in the treated population ranges from 53% to 75%.

92. The method of any one of claims 1 to 69, wherein a population of subjects is treated by the method and wherein the percentage of subjects with stable disease in the treated population is at least or about 22%.

93. The method of any one of claims 1 to 69, wherein a population of subjects is treated by the method, and wherein the duration of response in the treated population is at least or about 6 months.

94. The method of any one of claims 1 to 69, wherein a population of subjects is treated by the method, and wherein the duration of response in the treated population is at least or about 12 months.

95. The method of any one of claims 1 to 69, wherein a population of subjects is treated by the method, and wherein the duration of response in the treated population is at least or about 24 months.

96. The method of any one of claims 1 to 69, wherein a population of subjects is treated by the method, and wherein the duration of response in the treated population is at least or about 27 months.

97. The method of any one of claims 1 to 69, wherein a population of subjects is treated by the method, and wherein the progression-free survival in the treated population is at least or about 6 months.

98. The method of any one of claims 1 to 69, wherein a population of subjects is treated by the method, and wherein the progression-free survival in the treated population is at least or about 12 months.

99. The method of any one of claims 1 to 69, wherein a population of subjects is treated by the method, and wherein the progression-free survival in the treated population is at least or about 20 months.

100. The method of any one of claims 1 to 69, wherein a population of subjects is treated by the method, and wherein the progression-free survival in the treated population is at least or about 29 months.

101. The method of any one of claims 1 to 69, wherein a population of subjects is treated by the method, and wherein the median overall survival in the treated population is at least or about 22 months.

102. The method of any one of claims 1 to 69, wherein a population of subjects is treated by the method, and wherein the median overall survival in the treated population is at least or about 27 months.

103. The method of any one of claims 1 to 69, wherein a population of subjects is treated by the method, and wherein the median overall survival in the treated population is at least or about 30 months.

104. The method of any one of claims 1 to 69, wherein a population of subjects is treated by said method, and wherein the overall survival in the treated population ranges from 19 to 25 months.

105. The method of any one of claims 1 to 69, wherein a population of subjects is treated by said method, and wherein the overall survival in the treated population ranges from 30 to 32 months.

106. The method of any one of claims 1 to 70 and 72, wherein the complete response rate in a population of subjects treated with the method is at least or about 10%.

107. The method of any one of claims 1 to 69, 71 and 72, wherein the partial response rate in a population of subjects treated with the method is at least or about 54%.

108. The method of any one of claims 1 to 72, wherein the objective response rate in a population of subjects treated with the method is at least or about 65%.

109. The method of any one of claims 1 to 72, wherein the objective response rate in a population of subjects treated with said method is 53% to 75%.

110. The method of any one of claims 1 to 69 and 73, wherein the disease stabilization rate in a population of subjects treated with the method is at least or about 22%.

111. The method of any one of claims 1 to 69 and 74 to 77, wherein the duration of response in a population of subjects treated with the method is at least or about 6 months.

112. The method of any one of claims 1 to 69 and 74 to 77, wherein the duration of response in a population of subjects treated with the method is at least or about 12 months.

113. The method of any one of claims 1 to 69 and 74 to 77, wherein the duration of response in a population of subjects treated with the method is at least or about 24 months.

114. The method of any one of claims 1 to 69 and 74 to 77, wherein the duration of response in a population of subjects treated with the method is at least or about 27 months.

115. The method of any one of claims 1 to 69 and 78 to 82, wherein the progression-free survival of a population of subjects treated with the method is at least or about 6 months.

116. The method of any one of claims 1 to 69 and 78 to 82, wherein the progression-free survival of a population of subjects treated with the method is at least or about 12 months.

117. The method of any one of claims 1 to 69 and 78 to 82, wherein the progression-free survival of a population of subjects treated with the method is at least or about 20 months.

118. The method of any one of claims 1 to 69 and 78 to 82, wherein the progression-free survival of a population of subjects treated with the method is at least or about 29 months.

119. The method of any one of claims 1 to 69 and 83 to 87, wherein the median overall survival of a population of subjects treated with the method is at least or about 22 months.

120. The method of any one of claims 1 to 69 and 83 to 87, wherein the median overall survival of a population of subjects treated with the method is at least or about 27 months.

121. The method of any one of claims 1 to 69 and 83 to 87, wherein the median overall survival of a population of subjects treated with the method is at least or about 30 months.

122. The method of any one of claims 1 to 69, 79 and 80, wherein the overall survival of a population of subjects treated with said method is 19 to 25 months.

123. The method of any one of claims 1 to 69, 79 and 80, wherein the overall survival of a population of subjects treated with said method is 30 to 32 months.

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