Dosing regimen for anti-DLL3 agents

Abstract

The present invention provides a method for treating DLL3-positive cancer or SCLC, the method comprising administering an anti-DLL3 agent to a subject in need thereof once every two weeks at a dose of between about 0.3 mg and about 100 mg or between about 3 mg and about 200 mg. Also disclosed is a stepwise administration of an anti-DLL3 agent for treating SCLC.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of European Patent Application No. 19208214, filed on November 10, 2019, and U.S. Provisional Application No. 63 / 078,131, filed on September 14, 2020. The contents of both applications are incorporated herein by reference in their entirety.

[0003] Submit sequence listing as ASCII text file

[0004] The following content submitted in an ASCII text file is incorporated herein by reference in its entirety: Computer Readable Form (CRF) of the Sequence Listing (File Name: A-2520-WO-PCT_ST25.txt, Creation Date: October 28, 2020, Size: 1,218,887 bytes). Technical Field

[0005] The present application relates to dosage and administration of anti-DLL3 agents for treating cancer. Background Art

[0006] Small cell lung cancer (SCLC) is an aggressive form of lung cancer with a poor prognosis and limited treatment options, and represents approximately 10%-15% of lung cancers. For decades, survival rates have remained low, with only 5% of SCLC patients surviving five years, largely due to the lack of new therapies to combat this form of lung cancer. Approximately one-third of patients present with limited-stage disease. Most patients present with extensive-stage disease, defined as tumors present on only one side of the chest and which are amenable to a single field of radiation. These stages influence the available treatment options, with limited-stage disease being treated with chemotherapy and radiation and extensive-stage disease being treated with chemotherapy alone. Disseminated, metastatic tumors with lymphomatous features are the hallmark of SCLC.

[0007] Patients typically respond well to current first-line therapies, including etoposide and cisplatin, but chemoresistant disease invariably relapses quickly, for which there are currently no available treatment options. The prognosis for the relapsed refractory case is very poor, with rapid disease progression and a short median survival of less than six months. In addition, SCLC patients have a high rate of comorbidities, including hypertension, heart disease, diabetes, and tumor-associated syndromes. These, coupled with the typically advanced age of SCLC patients, affect the patient's ability to tolerate harsh chemotherapy regimens, further limiting treatment options.

[0008] Delta-like 3 (DLL3) is a type 1 transmembrane protein and non-classical Notch ligand that is differentially expressed in SCLC. Using immunohistochemistry (IHC), 85% of SCLC tumors stain positive for DLL3 (patterns consistent with membranous and cytoplasmic expression). In contrast, low levels of DLL3 protein expression are detected with a cytoplasmic staining pattern in normal brain, pancreatic islets, and pituitary (Saunders et al., Sci Transl Med. 7:302ra136 (2015)). DLL3 is a novel and promising target for the development of T cell-targeted therapies for SCLC.

[0009] There is an unmet medical need for the development of therapies to treat SCLC. Summary of the invention

[0010] Based on the disclosure provided herein, those skilled in the art will recognize, or be able to ascertain using only routine experimentation, many equivalent embodiments of the specific embodiments of the invention described herein. Such equivalent embodiments are intended to be encompassed by the following Example (E).

[0011] E1: A method of treating a DLL3-positive cancer, the method comprising administering an anti-DLL3 agent to a subject in need thereof, wherein the anti-DLL3 agent is administered at a dose of 0.3 mg to 100 mg, from 3 mg to 200 mg, or 100 mg once every two weeks.

[0012] E2: A method of treating a DLL3-positive cancer, the method comprising administering an anti-DLL3 agent to a subject in need thereof, wherein the anti-DLL3 agent is administered according to the following regimen: a) a first dose of from 0.3 mg to 100 mg, 0.5 mg to 10 mg, or 1 mg on day 1, b) a second dose of from 0.3 mg to 100 mg, from 3 to 200 mg, or 100 mg on day 8, and c) subsequent doses of one or more of from 0.3 mg to 100 mg, from 3 mg to 200 mg, or 100 mg starting on day 15 and every two weeks thereafter, and wherein the second dose and subsequent doses are the same and higher than the first dose.

[0013] E3: A method of treating a DLL3-positive cancer, the method comprising administering an anti-DLL3 agent to a subject in need thereof, wherein the anti-DLL3 agent is administered according to one of the following two regimens:

[0014] Regimen I: a) a first dose of 0.5 mg to 10 mg or 1 mg on day 1 (run-in dose), b) a second dose of 3 mg to 100 mg or 25 mg to 50 mg on day 4 (step dose), c) a third dose of 3 mg to 200 mg or 100 mg on day 8 (step dose), and d) one or more subsequent doses of 3 mg to 200 mg or 100 mg starting on day 15 and every two weeks thereafter (target dose), wherein the second dose is higher than the first dose, and the third dose and the subsequent dose are the same and are the same as or higher than the second dose; or

[0015] Regimen II: a) a first dose (run-in dose) from 0.5 mg to 10 mg or 1 mg on day 1, b) a second dose (step dose) from 3 mg to 100 mg or 25 mg to 50 mg on day 8, c) a third dose (step dose) from 3 mg to 200 mg or 100 mg on day 15, and c) one or more subsequent doses (target dose) from 3 mg to 200 mg or 100 mg starting on day 29 and every two weeks thereafter, and wherein the second dose is higher than the first dose, and the third dose and the subsequent dose are the same and higher than the second dose.

[0016] E4: A method of treating DLL3-positive cancer, the method comprising administering an anti-DLL3 agent to a subject in need thereof, wherein the anti-DLL3 agent is administered according to the following regimen: a) a first dose from 0.5 mg to 10 mg or 1 mg on day 1 (run-in dose), b) a second dose from 3 mg to 100 mg or 25 mg on day 4 (step dose), c) a third dose from 3 mg to 100 mg or 50 mg on day 8 (step dose), d) a fourth dose from 3 mg to 200 mg or 100 mg on day 15 (step dose), and e) one or more subsequent doses from 3 mg to 200 mg or 100 mg starting on day 29 and every two weeks thereafter (target dose), and wherein the second dose is higher than the first dose, the third dose is higher than the second dose, and the fourth dose and the subsequent dose are the same, and are the same as the third dose and higher than the third dose.

[0017] E5: A method for treating DLL3-positive cancer, the method comprising administering an anti-DLL3 agent to a subject in need thereof, wherein the anti-DLL3 agent is administered according to the following regimen: a) a first dose from 0.5 mg to 10 mg on day 1 (run-in dose), b) a second dose from 3 mg to 200 mg on day 4 (step dose), c) a third dose from 3 mg to 200 mg on day 8 (target dose), and d) one or more subsequent doses from 3 mg to 200 mg starting on day 15 and every two weeks thereafter (target dose), and wherein the second dose is higher than the first dose, and the third dose and the subsequent dose are the same, and are the same as the second dose.

[0018] E6: A method of treating DLL3-positive cancer, the method comprising administering an anti-DLL3 agent to a subject in need thereof, wherein the anti-DLL3 agent is administered according to the following regimen: a) a first dose from 0.5 mg to 10 mg on day 1 (run-in dose), b) a second dose from 3 mg to 100 mg on day 4 (step dose), c) a third dose from 3 mg to 200 mg on day 8 (step dose), d) a fourth dose from 3 mg to 200 mg on day 15 (target dose), and e) one or more subsequent doses from 3 mg to 200 mg starting on day 29 and every two weeks thereafter (target dose), and wherein the second dose is higher than the first dose, the third dose is higher than the second dose, and the fourth dose and the subsequent doses are the same, and are the same as the third dose.

[0019] E7: The method of any one of E1-E6, wherein the anti-DLL3-positive cancer is small cell lung cancer (SCLC).

[0020] E8: The method of any one of E1-E7, wherein the anti-DLL3-positive cancer is relapsed / refractory (RR) SCLC or extensive disease (ED) SCLC.

[0021] E9: The method of any one of E1-E8, wherein the anti-DLL3 agent is a bispecific antibody construct comprising two binding domains: the first domain binds to human DLL3, and the second domain binds to human CD3.

[0022] E10: The method of E9, wherein the DLL3 binding domain binds to an epitope of human DLL3 contained within the amino acid sequence of SEQ ID NO:258.

[0023] E11: A method as described in E9 or E10, wherein the DLL3 binding domain comprises (a) a heavy chain variable region (VH), the heavy chain variable region comprising: (i) VH complementarity determining region 1 (CDR-H1), which comprises the amino acid sequence of SEQ ID NO:31; (ii) CDR-H2, which comprises the amino acid sequence of SEQ ID NO:32; and (iii) CDR-H3, which comprises the amino acid sequence of SEQ ID NO:33; and (b) a light chain variable region (VL), the light chain variable region comprising: (i) VL complementarity determining region 1 (CDR-L1), which comprises the amino acid sequence of SEQ ID NO:34; (ii) CDR-L2, which comprises the amino acid sequence of SEQ ID NO:35; and (iii) CDR-L3, which comprises the amino acid sequence of SEQ ID NO:36.

[0024] E12: The method of any one of E9-E11, wherein the DLL3 binding domain comprises: (1) a VH comprising the amino acid sequence of SEQ ID NO:37 and a VL comprising the amino acid sequence of SEQ ID NO:38, or (2) a VH comprising the amino acid sequence of SEQ ID NO:435 and a VL comprising the amino acid sequence of SEQ ID NO:436.

[0025] E13: The method of any one of E9-E12, wherein the VH and VL of the DLL3 binding domain are connected by a linker to form a single-chain Fv (scFv).

[0026] E14: The method as described in E13, wherein the linker comprises a sequence selected from any one of SEQ ID NOs: 285-293.

[0027] E15: The method of E13 or E14, wherein the linker comprises (Gly4Ser)x, wherein x is an integer of 1 or greater (eg, 1, 2, 3 or 4).

[0028] E16: The method of any one of E9-E15, wherein the DLL3 binding domain comprises the amino acid sequence of SEQ ID NO:39 or SEQ ID NO:437.

[0029] E17: A method as described in any one of E9-E16, wherein the CD3 binding domain comprises: (a) VH, which comprises: CDR-H1 comprising the amino acid sequence of SEQ ID NO:426, CDR-H2 comprising the amino acid sequence of SEQ ID NO:427, and CDR-H3 comprising the amino acid sequence of SEQ ID NO:428; and VL, which comprises: CDR-L1 comprising the amino acid sequence of SEQ ID NO:423, CDR-L2 comprising the amino acid sequence of SEQ ID NO:424, and CDR-L3 comprising the amino acid sequence of SEQ ID NO:425.

[0030] E18: The method of any one of E9-E17, wherein the CD3 binding domain comprises: a VH comprising the amino acid sequence of SEQ ID NO: 429 and a VL comprising the amino acid sequence of SEQ ID NO: 430.

[0031] E19: The method of E17 or E18, wherein the VH and VL of the CD3 binding domain are connected by a linker to form a single-chain Fv (scFv).

[0032] E20: The method as described in E19, wherein the linker comprises a sequence selected from any one of SEQ ID NOs: 285-293.

[0033] E21: The method of E19 or E20, wherein the linker comprises (Gly4Ser)x, wherein x is an integer of 1 or greater (eg, 1, 2, 3 or 4).

[0034] E22: The method of any one of E17-E21, wherein the CD3 binding domain comprises the amino acid sequence of SEQ ID NO:431.

[0035] E23: The method of any one of E9-E22, wherein the DLL3 binding domain and the CD3 binding domain are connected by a linker.

[0036] E24: The method of E23, wherein the linker is a peptide linker comprising a sequence selected from any one of SEQ ID NOs: 285-293.

[0037] E25: The method of E23 or E24, wherein the linker is a peptide linker comprising (Gly4Ser)x, wherein x is an integer of 1 or greater (eg, 1, 2, 3 or 4).

[0038] E26: The method of any one of E9-E25, wherein the anti-DLL3 agent comprises a DLL3 binding domain and a CD3 binding domain. The DLL3 binding domain comprises (a) a heavy chain variable region (VH), the heavy chain variable region comprising: (i) a VH complementarity determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO: 31; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 32; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 33; and (b) a light chain variable region (VL), the light chain variable region comprising: (i) a VL complementarity determining region 1 (CDR-L1) comprising the amino acid sequence of SEQ ID NO: 34; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 35; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 36. The CD3 binding domain comprises (a) a VH comprising: (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 426, (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 427, and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 428; and (b) a VL comprising: (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 423, (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 424, and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 425.

[0039] E27: The method as described in any one of E9-E26, wherein the DLL3 binding domain comprises a VH comprising the amino acid sequence of SEQ ID NO:37 and a VL comprising the amino acid sequence of SEQ ID NO:38, and the CD3 binding domain comprises a VH comprising the amino acid sequence of SEQ ID NO:429 and a VL comprising the amino acid sequence of SEQ ID NO:430.

[0040] E28: The method as described in any one of E9-E26, wherein the DLL3 binding domain comprises a VH comprising the amino acid sequence of SEQ ID NO:435 and a VL comprising the amino acid sequence of SEQ ID NO:436, and the CD3 binding domain comprises a VH comprising the amino acid sequence of SEQ ID NO:429 and a VL comprising the amino acid sequence of SEQ ID NO:430.

[0041] E29: The method of any one of E9-E27, wherein the DLL3 binding domain comprises the amino acids of SEQ ID NO:39 and the CD3 binding domain comprises the amino acids of SEQ ID NO:431.

[0042] E30: The method of any one of E9-E26 or E28, wherein the DLL3 binding domain comprises the amino acids of SEQ ID NO: 437 and the CD3 binding domain comprises the amino acids of SEQ ID NO: 431.

[0043] E31: The method of E29, wherein the anti-DLL3 agent comprises the amino acid sequence of SEQ ID NO:40.

[0044] E32: The method of E30, wherein the anti-DLL3 agent comprises the amino acid sequence of SEQ ID NO:438.

[0045] E33: The method of any one of E9-E32, wherein the anti-DLL3 agent further comprises a third domain that prolongs or enhances the serum half-life of the anti-DLL3 agent.

[0046] E34: The method of E33, wherein the third domain comprises an amino acid sequence selected from any one of SEQ ID NOs: 541-548.

[0047] E35: The method of any one of E9-E26, E28, E30, E32, E33 or E35, wherein the anti-DLL3 agent comprises the amino acid of SEQ ID NO:520.

[0048] E36: The method of any one of E1 or E7-E35, wherein the anti-DLL3 agent is administered once every two weeks at a dose of from about 0.3 mg to about 90 mg, from about 0.3 mg to about 80 mg, from about 0.3 mg to about 70 mg, from about 0.3 mg to about 60 mg, from about 0.3 mg to about 50 mg, from about 0.3 mg to about 40 mg, from about 0.3 mg to about 30 mg, from about 0.3 mg to about 20 mg, from about 0.3 mg to about 10 mg, from about 0.3 mg to about 3 mg, from about 0.3 mg to about 10 ...50 mg, from about 0.3 mg to about 50 mg, from about 0.3 mg to about 40 mg, from about 0.3 mg to about from about 1 mg to about 100 mg, from about 1 mg to about 90 mg, from about 1 mg to about 80 mg, from about 1 mg to about 70 mg, from about 1 mg to about 60 mg, from about 1 mg to about 50 mg, from about 1 mg to about 40 mg, from about 1 mg to about 30 mg, from about 1 mg to about 20 mg, from about 1 mg to about 10 mg, from about 1 mg to about 3 mg, from about 3 mg to about 100 mg, from about 3 mg to about 90 mg, from about 3 mg to about 80 mg, from about 3 mg to about 70 mg, from about 3 mg to about 60 mg, from about 3 mg to about 50 mg. g, from about 3 mg to about 40 mg, from about 3 mg to about 30 mg, from about 3 mg to about 20 mg, from about 3 mg to about 10 mg, from about 3 mg to about 12 mg, from about 3 mg to about 15 mg, from about 10 mg to about 100 mg, from about 10 mg to about 90 mg, from about 10 mg to about 80 mg, from about 10 mg to about 70 mg, from about 10 mg to about 60 mg, from about 10 mg to about 50 mg, from about 10 mg to about 40 mg, from about 10 mg to about 30 mg, from about 10 mg to about 20 mg, from about 10 mg to about 1 From about 20 mg to about 100 mg, from about 20 mg to about 90 mg, from about 20 mg to about 80 mg, from about 20 mg to about 70 mg, from about 20 mg to about 60 mg, from about 20 mg to about 50 mg, from about 20 mg to about 40 mg, from about 20 mg to about 30 mg, from about 30 mg to about 100 mg, from about 30 mg to about 90 mg, from about 30 mg to about 80 mg, from about 30 mg to about 70 mg, from about 30 mg to about 60 mg, from about 30 mg to about 50 mg, or from about 30 mg to about 40 mg.

[0049] E37: The method of any one of E1 or E7-E35, wherein the anti-DLL3 agent is administered once every two weeks at a dose of from about 3 mg to about 100 mg, from about 10 to about 180 mg, from about 10 to about 150 mg, from about 30 mg to about 200 mg, from about 30 mg to about 180 mg, from about 30 mg to about 150 mg, from about 30 mg to about 120 mg, from about 50 mg to about 200 mg, from about 50 mg to about 180 mg, from about 50 mg to about 150 mg, from about 50 mg to about 120 mg, from about from about 70 mg to about 200 mg, from about 70 mg to about 180 mg, from about 70 mg to about 150 mg, from about 70 mg to about 120 mg, from about 90 mg to about 200 mg, from about 90 mg to about 180 mg, from about 90 mg to about 150 mg, from about 90 mg to about 120 mg, from about 100 mg to about 180 mg, from about 100 mg to about 150 mg, or from about 100 mg to about 120 mg; or wherein the anti-DLL3 agent is administered at a dose of 3 mg, 10 mg, 30 mg or 100 mg once every two weeks.

[0050] E38: The method of any one of E1 or E7-E37, wherein the anti-DLL3 agent is administered on days 1 and 15 of a 28-day cycle.

[0051] E39: A method as described in any of E2 or E7-E35, wherein the second dose and the one or more subsequent doses are the same and are at least 1.5 times, at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 20 times, at least 25 times, at least 30 times, at least 35 times, at least 40 times, at least 45 times, at least 50 times, at least 55 times, at least 60 times, at least 65 times, at least 70 times, at least 75 times, at least 80 times, at least 85 times, at least 90 times, at least 95 times, at least 100 times, at least 120 times, at least 150 times, at least 200 times higher than the first dose.

[0052] E40: The method of any one of E2, E7-E35 or E39, wherein each of the first doses of the anti-DLL3 agent is from about 0.5 mg to about 10 mg, from about 0.5 mg to about 8 mg, from about 0.5 mg to about 6 mg, from about 0.5 mg to about 4 mg, from about 0.5 mg to about 2 mg, or about 1 mg, and the second and subsequent doses of the anti-DLL3 agent can be any of: from about 0.3 mg to about 90 mg, from about 0.3 mg to about 80 mg, from about 0.3 mg to about 70 mg, from about 0.3 mg to about 60 mg, from about 0.3 mg to about 50 mg, from about 0.3 mg to about 40 mg, from about 0.3 mg to about 30 mg, from about 0.3 mg to about 20 mg, from about 0.3 mg to about 10 mg, from about 0.3 mg to about 30 mg, from about 0.3 mg to about 40 mg,3 mg to about 1 mg, from about 1 mg to about 100 mg, from about 1 mg to about 90 mg, from about 1 mg to about 80 mg, from about 1 mg to about 70 mg, from about 1 mg to about 60 mg, from about 1 mg to about 50 mg, from about 1 mg to about 40 mg, from about 1 mg to about 30 mg, from about 1 mg to about 20 mg, from about 1 mg to about 10 mg, from about 1 mg to about 3 mg, from about 3 mg to about 100 mg, from about 3 mg to about 90 mg, from about 3 mg to about 80 mg, from about 3 mg to about 70 mg, from about 3 mg to about 60 mg, from about 3 mg to about 50 mg, from about 3 mg to about 40 mg, from about 3 mg to about to about 30 mg, from about 3 mg to about 20 mg, from about 3 mg to about 10 mg, from about 3 mg to about 12 mg, from about 3 mg to about 15 mg, from about 10 mg to about 100 mg, from about 10 mg to about 90 mg, from about 10 mg to about 80 mg, from about 10 mg to about 70 mg, from about 10 mg to about 60 mg, from about 10 mg to about 50 mg, from about 10 mg to about 40 mg, from about 10 mg to about 30 mg, from about 10 mg to about 20 mg, from about 10 mg to about 15 mg, from about 20 mg to about 100 mg, from about 20 mg to about 90 mg, from about 20 mg to about 80 mg, from about 20 mg to about 70 mg, from about 20 mg to about 60 mg, from about 20 mg to about 50 mg, from about 20 mg to about 40 mg, from about 20 mg to about 30 mg, from about 30 mg to about 100 mg, from about 30 mg to about 90 mg, from about 30 mg to about 80 mg, from about 30 mg to about 70 mg, from about 30 mg to about 60 mg, from about 30 mg to about 50 mg, from about 30 mg to about 40 mg, from about 10 mg to about 180 mg, from about 10 mg to about 150 mg, from about 10 mg to about 120 mg, from about 30 mg to about 200 mg, from about 30 mg to about 180 mg, from about 30 mg to about 150 mg, from about 30 mg to about 120 mg, from about 50 mg to about 200 mg, from about 50 mg to about 180 mg, from about 50 mg to about 150 mg, from about 50 mg to about 120 mg, from about 70 mg to about 200 mg, from about 70 mg to about 180 mg, from about 70 mg to about 150 mg, from about 70 mg to about 120 mg, from about 90 mg to about 200 mg, from about 90 mg to about 180 mg, from about 90 mg to about 150 mg, from about 90 mg to about 120 mg, from about 100 mg to about 180 mg, from about 100 mg to about 150 mg, and from about 100 mg to about 120 mg. .

[0053] E41: The method of any one of E2, E7-E35, E39 or E40, wherein the first dose of the anti-DLL3 agent is 1 mg and the second and subsequent doses of the anti-DLL3 agent are each 3 mg, 10 mg, 30 mg or 100 mg.

[0054] E42: The method of any one of E3 or E7-E35, wherein the first dose of the anti-DLL3 agent is from about 0.5 mg to about 8 mg, from about 0.5 mg to about 6 mg, from about 0.5 mg to about 4 mg, from about 0.5 mg to about 2 mg, or 1 mg, the second dose of the anti-DLL3 agent is from about 3 mg to about 10 mg, from about 10 mg to about 100 mg, from about 10 mg to about 80 mg, from about 10 mg to about 60 mg, from about 20 mg to about 80 mg, from about 20 mg to about 60 mg, from about 25 mg to about 50 mg, from about 30 mg to about 80 mg, from about 30 mg to about 60 mg, from about 30 mg to about 40 mg, from about 40 mg to about 80 mg, or from about 40 mg to about 60 mg, and the third and subsequent doses of the anti-DLL3 agent can be any of the following: from about 3 mg to about 10 From about 10 mg to about 180 mg, from about 10 mg to about 150 mg, from about 10 mg to about 120 mg, from about 30 mg to about 200 mg, from about 30 mg to about 180 mg, from about 30 mg to about 150 mg, from about 30 mg to about 120 mg, from about 50 mg to about 200 mg, from about 50 mg to about 180 mg, from about 50 mg to about 150 mg, from about 70 mg to about 200 mg, from about 70 mg to about 180 mg, from about 70 mg to about 150 mg, from about 70 mg to about 120 mg, from about 90 mg to about 200 mg, from about 90 mg to about 180 mg, from about 90 mg to about 150 mg, from about 90 mg to about 120 mg, from about 100 mg to about 180 mg, from about 100 mg to about 150 mg, from about 100 mg to about 120 mg, or about 100 mg.

[0055] E43: The method of any one of E3, E7-E35 or E42, wherein the first dose of the anti-DLL3 agent is 1 mg, the second dose of the anti-DLL3 agent is from 25 mg to 50 mg or from 45 mg to 70 mg, and the third and subsequent doses of the anti-DLL3 agent are 100 mg.

[0056] E44: A method as described in any of E3, E7-E35 or E42, wherein the first dose of the anti-DLL3 agent on day 1 is 1 mg, the second dose of the anti-DLL3 agent on day 4 is 25 mg or 50 mg, the third dose of the anti-DLL3 agent on day 8 is 100 mg, and starting from day 15 and once every two weeks thereafter, the subsequent dose of the anti-DLL3 agent is 100 mg.

[0057] E45: The method of any one of E4 or E7-E35, wherein the first dose of the anti-DLL3 agent is from about 0.5 mg to about 8 mg, from about 0.5 mg to about 6 mg, from about 0.5 mg to about 4 mg, from about 0.5 mg to about 2 mg, or 1 mg, the second dose of the anti-DLL3 agent is from about 3 mg to about 10 mg, from about 10 mg to about 100 mg, from about 10 mg to about 80 mg, from about 10 mg to about 60 mg, from about 10 mg to about 40 mg, from about 20 mg to about 80 mg, from about 20 mg to about 60 mg, from about 20 mg to about 40 mg, or about 25 mg, and the third dose of the anti-DLL3 agent is From about 3 mg to about 10 mg, from about 10 mg to about 100 mg, from about 10 mg to about 80 mg, from about 10 mg to about 60 mg, from about 20 mg to about 100 mg, from about 20 mg to about 80 mg, from about 20 mg to about 60 mg, from about 30 mg to about 100 mg, from about 30 mg to about 80 mg, from about 30 mg to about 60 mg, from about 40 mg to about 100 mg, from about 40 mg to about 80 mg, from about 40 mg to about 60 mg, or about 50 mg, the fourth and subsequent doses of the anti-DLL3 agent may be any of the following: from about 3 mg to about 100 mg, from about 10 mg to about 150 mg, from about 30 mg to about 100 mg, from about 20 mg to about 80 mg, from about 20 mg to about 60 mg, From about 30 mg to about 150 mg, from about 30 mg to about 100 mg, from about 50 mg to about 200 mg, from about 50 mg to about 180 mg, from about 50 mg to about 150 mg, from about 50 mg to about 120 mg, from about 70 mg to about 200 mg, from about 70 mg to about 180 mg, from about 70 mg to about 120 mg, from about 90 mg to about 200 mg, from about 90 mg to about 180 mg, from about 90 mg to about 150 mg, from about 90 mg to about 120 mg, or about 100 mg.

[0058] E46: The method of any one of E4, E7-E35 or E45, wherein the first dose of the anti-DLL3 agent is 1 mg, the second dose of the anti-DLL3 agent is 25 mg, the third dose of the anti-DLL3 agent is 50 mg, and the fourth and subsequent doses of the anti-DLL3 agent are 100 mg.

[0059] E47: A method as described in any of E5 or E7-E35, wherein the first dose of the anti-DLL3 agent is 0.5 mg to 8 mg, from 0.5 mg to 6 mg, from 0.5 mg to 4 mg, from 0.5 mg to 2 mg, or 1 mg, and the second, third and subsequent doses are each from 3 mg to 100 mg, from 10 mg to 150 mg, from 30 mg to 200 mg, from 30 mg to 100 mg, from 50 mg to 150 mg, from 70 mg to 120 mg, from 90 mg to 120 mg, from 100 mg to 200 mg, or 100 mg.

[0060] E48: A method as described in any of E6 or E7-E35, wherein the first dose is from 0.5 mg to 8 mg, from 0.5 mg to 6 mg, from 0.5 mg to 4 mg, from 0.5 mg to 2 mg, or 1 mg, the second dose is from 3 mg to 10 mg, from 10 mg to 80 mg, from 10 mg to 60 mg, from 20 mg to 80 mg, from 20 mg to 60 mg, from 20 mg to 40 mg, 25 mg, or 50 mg, and the third, fourth and subsequent doses are each from 3 mg to 100 mg, from 10 mg to 150 mg, from 30 mg to 200 mg, from 30 mg to 100 mg, from 50 mg to 150 mg, from 70 mg to 120 mg, from 90 mg to 120 mg, 100 mg to 200 mg, or 100 mg.

[0061] E49: The method of any one of E1-E48, wherein the method comprises administering one or more additional therapeutic agents to the subject.

[0062] E50: The method of E49, wherein the one or more additional therapeutic agents is a steroid.

[0063] E51: The method of any of E49 or E50, wherein the additional therapeutic agent is dexamethasone.

[0064] E52: The method of any one of E49-E51, wherein the additional therapeutic agent is administered prior to treatment with the anti-DLL3 agent.

[0065] E53: The method of any one of E48-E51, wherein the additional therapeutic agent is administered concurrently with the anti-DLL3 agent.

[0066] E54: The method of any one of E2-E53, wherein the anti-DLL3 agent is administered in a 28-day cycle, and the method further comprises administering a fluid (e.g., saline), an anti-inflammatory agent, tocilizumab, or etanercept to the subject during the first cycle of administering the anti-DLL3 agent.

[0067] E55: The method of E54, wherein after the run-in dose and the step dose of the anti-DLL3 agent, one liter of saline is administered to the subject by intravenous infusion.

[0068] E56: The method of E55, wherein one liter of saline is administered over about 4-5 hours.

[0069] E57: The method according to E54, wherein the anti-inflammatory agent is a corticosteroid or acetaminophen.

[0070] E58: The method of E57, wherein the corticosteroid is dexamethasone.

[0071] E59: The method of any one of E54, E57 or E58, wherein an anti-inflammatory agent or tocilizumab is administered to the subject prior to the run-in dose and the stepped dose of the anti-DLL3 agent.

[0072] E60: The method of E59, wherein a corticosteroid is administered to the subject about 6-16 hours before the run-in dose and the stepped dose of the anti-DLL3 agent.

[0073] E61: The method of E59, wherein tocilizumab or acetaminophen is administered to the subject about one hour before the run-in dose and the stepped dose of the anti-DLL3 agent.

[0074] E62: The method of E54, wherein etanercept is administered to the subject about 36-60 hours before the run-in dose and the step doses of the anti-DLL3 agent (except the step dose on day 4).

[0075] E63: The method of E62, wherein etanercept is administered 2 days before the run-in dose and the step dose (except the step dose on day 4) of the anti-DLL3 agent.

[0076] E64: The method of any one of E1-E63, wherein the subject is a human.

[0077] E65: An anti-DLL3 agent for use in the method as described in any one of embodiments E1-E64.

[0078] E66: An anti-DLL3 agent for use in treating a DLL3-positive cancer (eg, SCLC), wherein the anti-DLL3 agent is administered as described in any one of Examples E1-E64.

[0079] E67: Use of an anti-DLL3 agent for the preparation of a medicament for the treatment of SCLC, wherein the medicament is to be administered as described in any one of Examples E1-E64.

[0080] E68: Use of an anti-DLL3 agent in the preparation of a medicament for the treatment of a DLL3-positive cancer, wherein the anti-DLL3 agent is administered as described in any one of Examples E1-E64. BRIEF DESCRIPTION OF THE DRAWINGS

[0081] Figure 1 Shown is the predicted serum concentration-time curve of AMG 757 in humans for a single cycle after administration of a short intravenous infusion once every two weeks. The dashed lines represent the concentrations required for 50% and 90% maximal effects of AMG 757-mediated CD69 upregulation (EC50 and EC90 values ​​of 0.61 ng / mL and 4.6 ng / mL, respectively), which were identified as the most sensitive marker of AMG757 activity (Study 123564).

[0082] Figure 2 Shown is the predicted lung concentration-time curve of AMG 757 in humans for a single cycle following administration of a short intravenous infusion once every two weeks. The dashed lines represent the concentrations required for 50% and 90% maximal effect of AMG 757-mediated cell killing in SHP-77 cells (EC50 and EC90 values ​​are 2.8 ng / mL and 5.7 ng / mL, respectively).

[0083] Figure 3 An example of stepped dosing is shown, assuming that adverse events associated with the first dose effect occur at 0.03 mg.

[0084] Figure 4 Confirmed partial responses (PR) in SCLC patients treated with AMG 757 were shown.

[0085] Figure 5 Shown are the mean steady-state serum AMG 757 concentration-time curves for SCLC patients treated with the molecule. Figure 5 In the table, * indicates stepped dosing, and data are shown during the 2-week dosing interval after the Day 15 dose in Cycle 2. Data from the only patient enrolled in the 0.1 mg group were not available because the patient withdrew during Cycle 1.

[0086] Figure 6 Showing a summary of objective responses in SCLC patients treated with AMG 757. DETAILED DESCRIPTION

[0087] As disclosed and exemplified herein, a Phase 1 clinical study was conducted for the treatment of SCLC using a bispecific protein targeting DLL3 and CD3 (AMG 757).

[0088] AMG 757 is a half-life-extended drug developed for the treatment of SCLC (bispecific T cell engager) molecule. The activity of AMG 757 requires simultaneous binding to target cells (DLL3 + AMG 757 has a pharmacological effect through previously primed cytotoxic CD8 + or CD4 + Specific redirection of T lymphocytes to mediate killing by DLL3 + The starting dose for the first-in-human (FIH) study was selected based on the minimum anticipated biological effect level (MABEL), which was identified as the EC for AMG 757-mediated CD69 upregulation in SHP-77 cells. 50 (Study 123564). Based on human PK predictions, a starting dose of 0.003 mg once every two weeks (Q2W) was selected, which is expected to produce a maximum serum concentration equivalent to MABEL (0.61 ng / mL). This regimen is expected to achieve adequate exposure to target tissues (such as the lungs) throughout the dosing interval while minimizing the peak-to-trough ratio after multiple treatment cycles of AMG 757. Based on clinical experience from the FIH study, the expected dose is at least 0.3 mg Q2W.

[0089] 1. Definition

[0090] Some exemplary bispecific anti-DLL3 agents disclosed herein (e.g. The antigen binding fragment (molecule) is a recombinant protein construct comprising two binding domains, each domain being derived from an antigen binding fragment of a full-length antibody. Such antigen binding fragments retain the ability to specifically bind to an antigen (preferably with substantially the same binding affinity). Examples of antigen binding fragments include (i) a Fab fragment, a monovalent fragment consisting of a VL, VH, CL, and CH1 domain; (ii) a F(ab')2 fragment, which is a bivalent fragment comprising two Fab fragments connected by a disulfide bridge at the hinge region; (iii) a Fd fragment consisting of a VH and CH1 domain; (iv) a Fv fragment consisting of the VL and VH domains of a single arm of an antibody, and (v) a dAb fragment (Ward et al., 1989 Nature 341: 544-546), which consists of a VH domain. In addition, although the two domains of the Fv fragment, VL and VH, are encoded by separate genes, recombinant methods can be used to connect the two domains through a synthetic linker that enables them to form a single protein chain, in which the VL region and the VH region pair to form a monovalent molecule (called a single-chain Fv (scFv); see, e.g., Bird et al. Science 242:423-426 (1988) and Huston et al., 1988, Proc. Natl. Acad. Sci. USA 85:5879-5883).

[0091] "Variable domain" refers to the variable region of an antibody light chain (VL) or the variable region of an antibody heavy chain (VH), whether alone or in combination. As known in the art, the variable regions of the heavy and light chains are each composed of four framework regions (FR) connected by three complementarity determining regions (CDRs), and contribute to the formation of the antigen binding site of an antibody.

[0092] "Complementarity determining regions" (CDRs) of exemplary DLL3 binding domains and CD3 binding domains are provided in the sequence listing. CDRs can be defined according to the Kabat, Chothia, cumulative, AbM, contact, North, and / or conformational definitions of Kabat and Chothia, or any CDR determination method well known in the art. See, e.g., Kabat et al., 1991, Sequences of Proteins of Immunological Interest, 5th Ed. (hypervariable regions); Chothia et al., 1989, Nature 342:877-883 (structural loop structures). The AbM definition of CDRs is a compromise between Kabat and Chothia and uses the AbM antibody modeling software from Oxford Molecular. The identity of the amino acid residues in a particular antibody that comprise a CDR can be determined using methods well known in the art.

[0093] The term "treatment" includes prophylactic and / or therapeutic treatment. If administered prior to clinical manifestation of a disorder, the treatment is considered prophylactic. Therapeutic treatment includes, for example, ameliorating or reducing the severity of a disease, or shortening the length of a disease.

[0094] "About" or "approximately," when used in conjunction with a measurable numerical variable, refers to the indicated value of the variable and all values ​​of the variable that are within the experimental error range of the indicated value (e.g., within a 95% confidence interval for the mean) or ±10% of the indicated value, whichever is greater. Numerical ranges are inclusive of the numbers defining the range.

[0095] A "run-in dose" when used in connection with administration of an anti-DLL3 agent for the treatment of cancer (e.g., small cell lung cancer (SCLC)) refers to an initial dose of the anti-DLL3 agent that is equal to or lower than the dose at which a first dose effect (e.g., cytokine release syndrome (CRS)) is observed. As is known in the art, a run-in dose can be determined by modeling and simulation of safety and pharmacokinetic data. For example, a run-in dose can be the maximum tolerated dose (MTD) of the anti-DLL3 agent at which no CRS or less than a certain grade (e.g., Grade 2) of CRS is observed.

[0096] "Target dose" when referring to the administration of an anti-DLL3 agent for treating cancer (eg, SCLC) refers to the dose that achieves the target effect of the anti-DLL3 agent (eg, improving or reducing the severity of SCLC, or shortening the length of SCLC).

[0097] "Step dose" when referring to the administration of an anti-DLL3 agent for the treatment of cancer (eg, SCLC) refers to a dose that is higher than the dose of the previously administered anti-DLL3 agent. Step dose includes one or more doses that increase from a run-in dose to reach a target dose.

[0098] 2. Anti-DLL3 Agents

[0099] DLL3 is an atypical Notch ligand that is primarily expressed during embryonic development (playing a role in somitogenesis). Compared to other Notch ligands expressed on the cell surface, DLL3 accumulates in the Golgi apparatus in normal tissues (Geffers et al., J Cell Biol. [Journal of Cell Biology] 178: 465-476 (2007)). By analyzing the differential expression of DLL3 in 28 SCLC tumors and a large number of normal tissues, DLL3 was identified as a tumor-associated antigen and an attractive target for T cell-based therapy (Study 123658).

[0100] The human DLL3 protein contains eight extracellular domains: signal peptide, N-terminus, DSL, EGF1, EGF2, EGF3, EGF4, EGF5 and EGF6. The amino acid sequences of human DLL3, EGF3 domain, EGF4 domain and combined EGF3 and EGF4 domains are shown in the sequence listing as SEQ ID NOs: 252, 258, 259 and 260, respectively.

[0101] Exemplary anti-DLL3 agents are bispecific molecules that bind DLL3 and CD3, e.g. (Bispecific T cell engager) molecules. The molecule is a recombinant protein construct made of two flexibly linked binding domains, each derived from an antibody. One binding domain of the molecule is specific for a tumor-associated surface antigen (e.g., DLL3); the second binding domain is specific for CD3, a subunit of the T cell receptor complex on T cells. The antibody constructs are uniquely suited to transiently link T cells to target cells and simultaneously strongly activate the intrinsic cytolytic potential of T cells against target cells. See, for example, WO 99 / 54440, WO 2005 / 040220, and WO 2008 / 119567.

[0102] Thus, in some embodiments, the anti-DLL3 agent comprises two binding domains: a first domain binds to DLL3 (preferably human DLL3), and a second domain binds to CD3 (preferably human CD3). Preferably, the first domain binds to an epitope of DLL3 contained within the amino acid sequence of SEQ ID NO: 260. More preferably, the first domain binds to an epitope of DLL3 contained within the amino acid sequence of SEQ ID NO: 258.

[0103] In certain embodiments, the DLL3 binding domain comprises (a) a heavy chain variable region (VH), comprising: (i) a VH complementarity determining region 1 (CDR-H1) comprising an amino acid sequence of SEQ ID NO:31; (ii) a CDR-H2 comprising an amino acid sequence of SEQ ID NO:32; and (iii) a CDR-H3 comprising an amino acid sequence of SEQ ID NO:33; and (b) a light chain variable region (VL), comprising: (i) a VL complementarity determining region 1 (CDR-L1) comprising an amino acid sequence of SEQ ID NO:34; (ii) a CDR-L2 comprising an amino acid sequence of SEQ ID NO:35; and (iii) a CDR-L3 comprising an amino acid sequence of SEQ ID NO:36.

[0104] In certain embodiments, the DLL3 binding domain comprises: a VH comprising the amino acid sequence of SEQ ID NO: 37 and a VL comprising the amino acid sequence of SEQ ID NO: 38. In certain preferred embodiments, the DLL3 binding domain comprises: a VH comprising the amino acid sequence of SEQ ID NO: 435 and a VL comprising the amino acid sequence of SEQ ID NO: 436.

[0105] In some embodiments, VH and VL are connected by a linker to form a single-chain Fv (scFv). In some embodiments, the linker is a peptide linker comprising a sequence selected from any one of SEQ ID NOs: 285-293. In some embodiments, the linker is a GS linker, such as Gly-Gly-Gly-Gly-Ser (G4S, SEQ ID NO: 286) or a polymer thereof, i.e. (Gly4Ser)x, wherein x is an integer of 1 or greater (e.g., 2 or 3) (e.g., SEQ ID NOs: 292, 293).

[0106] In certain embodiments, the DLL3 binding domain comprises the amino acid sequence of SEQ ID NO: 39. In certain preferred embodiments, the DLL3 binding domain comprises the amino acid sequence of SEQ ID NO: 437.

[0107] In certain embodiments, the CD3 binding domain comprises: (a) a VH comprising: a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 426, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 427, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 428; and a VL comprising: a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 423, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 424, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 425.

[0108] In certain embodiments, the CD3 binding domain comprises: a VH comprising the amino acid sequence of SEQ ID NO: 429 and a VL comprising the amino acid sequence of SEQ ID NO: 430. In some embodiments, VH and VL are connected by a linker to form a single-chain Fv (scFv). In some embodiments, the linker is a peptide linker comprising a sequence selected from any one of SEQ ID NOs: 285-293. In some embodiments, the linker is a GS linker, such as Gly-Gly-Gly-Gly-Ser (G4S, SEQ ID NO: 286) or a polymer thereof, i.e. (Gly4Ser)x, wherein x is an integer of 1 or greater (e.g., 2 or 3).

[0109] In certain embodiments, the CD3 binding domain comprises the amino acid sequence of SEQ ID NO:431.

[0110] In certain embodiments, the DLL3 binding domain and the CD3 binding domain are connected by a linker. In some embodiments, the linker is a peptide linker comprising a sequence selected from any one of SEQ ID NOs: 285-293. In some embodiments, the linker is a GS linker, such as Gly-Gly-Gly-Gly-Ser (G4S, SEQ ID NO: 286) or a polymer thereof, i.e., (Gly4Ser)x, wherein x is an integer of 1 or greater (e.g., 2 or 3).

[0111] In certain embodiments, the anti-DLL3 agents disclosed herein comprise two domains. The first domain binds to DLL3 (preferably human DLL3) and comprises (a) a heavy chain variable region (VH) comprising: (i) a VH complementarity determining region 1 (CDR-H1) comprising the amino acid sequence of SEQ ID NO: 31; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 32; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 33; and (b) a light chain variable region (VL) comprising: (i) a VL complementarity determining region 1 (CDR-L1) comprising the amino acid sequence of SEQ ID NO: 34; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 35; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 36. The second domain binds to CD3 (preferably human CD3) and comprises (a) a VH comprising: (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 426, (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 427, and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 428; and (b) a VL comprising: (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 423, (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 424, and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 425.

[0112] In certain embodiments, the anti-DLL3 agents described herein comprise two domains: (a) the first domain binds to DLL3 (preferably human DLL3) and comprises: a VH comprising the amino acid sequence of SEQ ID NO: 37 and a VL comprising the amino acid sequence of SEQ ID NO: 38; and (b) the second domain binds to CD3 (preferably human CD3) and comprises: a VH comprising the amino acid sequence of SEQ ID NO: 429 and a VL comprising the amino acid sequence of SEQ ID NO: 430. In certain preferred embodiments, the anti-DLL3 agents described herein comprise two domains: (a) the first domain binds to DLL3 (preferably human DLL3) and comprises: a VH comprising the amino acid sequence of SEQ ID NO: 435 and a VL comprising the amino acid sequence of SEQ ID NO: 436; and (b) the second domain binds to CD3 (preferably human CD3) and comprises: a VH comprising the amino acid sequence of SEQ ID NO: 429 and a VL comprising the amino acid sequence of SEQ ID NO: 430.

[0113] In certain embodiments, the anti-DLL3 agents described herein comprise two domains: (a) the first domain binds to DLL3 (preferably human DLL3) and comprises the amino acid sequence of SEQ ID NO: 39, (b) the second domain binds to CD3 (preferably human CD3) and comprises the amino acid sequence of SEQ ID NO: 431. In certain embodiments, the anti-DLL3 agents described herein comprise two domains: (a) the first domain binds to DLL3 (preferably human DLL3) and comprises the amino acid sequence of SEQ ID NO: 437, (b) the second domain binds to CD3 (preferably human CD3) and comprises the amino acid sequence of SEQ ID NO: 431.

[0114] In certain embodiments, an anti-DLL3 agent described herein comprises the amino acid sequence of SEQ ID NO: 40. In certain embodiments, an anti-DLL3 agent described herein comprises the amino acid sequence of SEQ ID NO: 438.

[0115] In certain embodiments, the anti-DLL3 agents described herein further comprise a third domain that prolongs or enhances the serum half-life of the anti-DLL3 agent. In certain embodiments, the third domain comprises two polypeptides connected by a linker, each peptide comprising the hinge, CH2, and CH3 domains of human IgG. In certain embodiments, the third domain comprises, in order from N-terminus to C-terminus: hinge-CH2-CH3-linker-hinge-CH2-CH3. In some embodiments, the linker is a GS linker, such as Gly-Gly-Gly-Gly-Ser (G4S, SEQ ID NO: 286) or a polymer thereof, i.e., (Gly4Ser)x, wherein x is an integer of 1 or greater (e.g., 6). In certain embodiments, the third domain comprises an amino acid sequence selected from any one of SEQ ID NOs: 541-548.

[0116] In certain embodiments, the DLL3 binding domain and the CD3 binding domain are connected by a first linker to form a peptide, which is connected to the third domain by a second linker. In certain embodiments, the first linker is a peptide linker comprising a sequence selected from any one of SEQ ID NOs: 285-293, and the second linker comprises a sequence selected from any one of SEQ ID NOs: 285, 286, 288, 289, 290, 292, and 293. In some embodiments, the first linker is a GS linker, such as Gly-Gly-Gly-Gly-Ser (G4S, SEQ ID NO: 286) or a polymer thereof, i.e., (Gly4Ser)x, wherein x is an integer of 1 or greater (e.g., 2 or 3) and the second linker comprises a sequence selected from any one of SEQ ID NOs: 285, 286, 288, 289, 290, 292, and 293.

[0117] In certain embodiments, the anti-DLL3 agents described herein comprise three domains: (a) the first domain binds to DLL3 (preferably human DLL3) and comprises the amino acid sequence of SEQ ID NO: 39, (b) the second domain binds to CD3 (preferably human CD3) and comprises the amino acid sequence of SEQ ID NO: 431, and (c) the third domain comprises an amino acid sequence selected from any one of SEQ ID NOs: 541-548. In certain embodiments, the anti-DLL3 agents described herein comprise three domains: (a) the first domain binds to DLL3 (preferably human DLL3) and comprises the amino acid sequence of SEQ ID NO: 437, (b) the second domain binds to CD3 (preferably human CD3) and comprises the amino acid sequence of SEQ ID NO: 431, and (c) the third domain comprises an amino acid sequence selected from any one of SEQ ID NOs: 541-548.

[0118] In certain embodiments, an anti-DLL3 agent described herein comprises the amino acid sequence of SEQ ID NO:520.

[0119] 3. Administration of Anti-DLL3 Agents

[0120] Disclosed herein are methods of treating a DLL3-positive cancer comprising administering an anti-DLL3 agent to a subject in need thereof at a dose of from about 0.3 mg to about 100 mg, from about 3 mg to about 200 mg, or about 100 mg, once every two weeks. In certain embodiments, the DLL3-positive cancer is small cell lung cancer (SCLC). In certain embodiments, the SCLC is relapsed / refractory SCLC (RR SCLC) or extensive disease SCLC (ED SCLC). In certain embodiments, the subject is a human with SCLC (e.g., RR SCLC or ED SCLC).

[0121] In certain embodiments, the anti-DLL3 agent is administered once every two weeks at a dose of from about 0.3 mg to about 100 mg, from about 0.3 mg to about 90 mg, from about 0.3 mg to about 80 mg, from about 0.3 mg to about 70 mg, from about 0.3 mg to about 60 mg, from about 0.3 mg to about 50 mg, from about 0.3 mg to about 40 mg, from about 0.3 mg to about 30 mg, from about 0.3 mg to about 20 mg, from about 0.3 mg to about 10 mg, from about 0.3 mg to about 30 mg, from about 0.3 mg to about 20 mg, from about 0.3 mg to about 10 mg, from about 0.3 mg to about 10 mg, from about 0.3 mg to about 10 mg. to about 100 mg, from about 1 mg to about 90 mg, from about 1 mg to about 80 mg, from about 1 mg to about 70 mg, from about 1 mg to about 60 mg, from about 1 mg to about 50 mg, from about 1 mg to about 40 mg, from about 1 mg to about 30 mg, from about 1 mg to about 20 mg, from about 1 mg to about 10 mg, from about 1 mg to about 3 mg, from about 3 mg to about 100 mg, from about 3 mg to about 90 mg, from about 3 mg to about 80 mg, from about 3 mg to about 70 mg, from about 3 mg to about 60 mg, from about 3 mg to about 50 mg, From about 3 mg to about 40 mg, from about 3 mg to about 30 mg, from about 3 mg to about 20 mg, from about 3 mg to about 10 mg, from about 3 mg to about 12 mg, from about 3 mg to about 15 mg, from about 10 mg to about 100 mg, from about 10 mg to about 90 mg, from about 10 mg to about 80 mg, from about 10 mg to about 70 mg, from about 10 mg to about 60 mg, from about 10 mg to about 50 mg, from about 10 mg to about 40 mg, from about 10 mg to about 30 mg, from about 10 mg to about 20 mg, from about 10 mg to about 15 From about 20 mg to about 100 mg, from about 20 mg to about 90 mg, from about 20 mg to about 80 mg, from about 20 mg to about 70 mg, from about 20 mg to about 60 mg, from about 20 mg to about 50 mg, from about 20 mg to about 40 mg, from about 20 mg to about 30 mg, from about 30 mg to about 100 mg, from about 30 mg to about 90 mg, from about 30 mg to about 80 mg, from about 30 mg to about 70 mg, from about 30 mg to about 60 mg, from about 30 mg to about 50 mg, or from about 30 mg to about 40 mg.

[0122] In certain embodiments, the anti-DLL3 agent is administered biweekly at a dose of from about 3 mg to about 100 mg, from about 10 mg to about 200 mg, from about 10 mg to about 180 mg, from about 10 mg to about 150 mg, from about 10 mg to about 120 mg, from about 30 mg to about 200 mg, from about 30 mg to about 180 mg, from about 30 mg to about 150 mg, from about 30 mg to about 120 mg, from about 50 mg to about 200 mg, from about 50 mg to about 180 mg, from about From about 50 mg to about 150 mg, from about 50 mg to about 120 mg, from about 70 mg to about 200 mg, from about 70 mg to about 180 mg, from about 70 mg to about 150 mg, from about 70 mg to about 120 mg, from about 90 mg to about 200 mg, from about 90 mg to about 180 mg, from about 90 mg to about 150 mg, from about 90 mg to about 120 mg, from about 100 mg to about 180 mg, from about 100 mg to about 150 mg, or from about 100 mg to about 120 mg.

[0123] In certain embodiments, the anti-DLL3 agent is administered at a dose of about 3 mg, 10 mg, 30 mg, or 100 mg once every two weeks.

[0124] In certain embodiments, the anti-DLL3 agent is administered on Days 1 and 15 of a 28-day cycle.

[0125] As described in Example 2, the starting dose of the first human (FIH) study is based on the minimum expected biological effect level (MABEL), which is identified as the EC50 of CD69 upregulation in SHP-77 cells mediated by AMG 757 (study 123564). Based on human PK predictions, a starting dose of 0.003 mg is selected, which is expected to produce a maximum serum concentration (0.61 ng / mL) equivalent to MABEL. In a Phase 1 clinical study, the dose of AMG 757 is 0.003 mg and higher, and is administered as an intravenous (IV) infusion once every two weeks (Q2W) in patients with SCLC. It is believed that this regimen achieves sufficient exposure to target tissues (such as lungs) throughout the dosing interval while minimizing the peak-to-trough ratio after multiple treatment cycles of AMG 757.

[0126] The predicted human PK parameters were used to predict AMG 757 concentrations in the lung, which was designated as a representative site of action and assumed to achieve approximately 1% of serum exposure (Vugmeyster et al., 2010). The mean EC values ​​for cell killing in SHP-77 cells were 90 Trough coverage (assuming 1% lung exposure) at 10 mg every 2 weeks predicts early signs of efficacy, see Example 2.

[0127] In clinical studies, there was a confirmed response to the administration of 0.3 mg (cohort 5) by intravenous infusion once every two weeks. This observed efficacy of AMG 757 is surprising because it is significantly lower than the dose of 10 mg Q2W predicted to be effective based on preclinical results. Additional confirmed responses were observed in the high-dose cohort. It is believed that the effective dose of AMG 757 can be at least 0.3 mg (e.g., from about 0.3 mg to about 100 mg or from about 3 mg to about 200 mg) administered once every two weeks.

[0128] The anti-DLL3 agent can be administered by any suitable means, including parenteral, subcutaneous, intraperitoneal, intrapulmonary, intranasal and / or intralesional administration. Parenteral administration includes intramuscular, intravenous, intraarterial, intraperitoneal or subcutaneous administration. In some embodiments, the anti-DLL3 agent is administered once every two weeks by intravenous (IV) infusion, such as a short intravenous infusion (approximately 60 minutes).

[0129] 3.1 Step-by-step dosing

[0130] Due to its mechanism of action, after the initial infusion of AMG 757, the risk of the subject's first dose effect (such as cytokine release syndrome) may increase. It is believed that the optimal maximum tolerated dose (MTD) may require a step-by-step dosing method (e.g., the initial dose on day 1, the step-by-step dose on day 8, and the subsequent doses starting on day 15 and thereafter). In some aspects, two MTDs can be estimated or established, one for initial dosing (MTD1, run-in dose) and one for subsequent dosing (MTD2). In some aspects, multiple MTDs can be estimated or established, one for the initial run-in dose (MTD1) and one for each one or more step doses and subsequent doses (if applicable).

[0131] If the subject experiences a first dose effect (e.g., cytokine release syndrome (CRS)), an appropriate first dose (MTD1) not exceeding the dose at which the CRS event is observed may be determined and implemented. A second dose and subsequent doses may also be determined and implemented. In addition, according to the number of steps in the step-dosing regimen, a second dose, a third dose, a fourth dose, and subsequent doses may be determined and implemented. In certain embodiments, the second dose is the same as the subsequent dose and is higher than the first dose. In certain embodiments, the second dose is higher than the first dose and the third dose is the same as the subsequent dose and is higher than the second dose. In certain embodiments, the second dose is higher than the first dose, the third dose is higher than the second dose, and the fourth dose is the same as the subsequent dose and is higher than the third dose. Due to the increased frequency of administration of the step-dosing regimen and possible drug accumulation, these doses and dosing regimens may be guided by modeling and simulation of clinical data (such as pharmacokinetics, safety data, etc.) to ensure that the systemic exposure of AMG757 does not exceed the dose associated with the observed first dose effect (such as CRS).

[0132] The table below shows an exemplary step-dosing schedule for an anti-DLL3 agent (eg, AMG 757) in a 28-day cycle (cycle 1 only), with the anti-DLL3 agent administered every two weeks thereafter.

[0133] Exemplary Single and Multiple Step Dosing Regimens (Cycle 1 Only)

[0134]

[0135] Accordingly, disclosed herein are methods of treating a DLL3-positive cancer, the methods comprising administering an anti-DLL3 agent to a subject in need thereof, wherein the anti-DLL3 agent is administered according to a step-dosing regimen.

[0136] In certain embodiments, disclosed herein is a method of treating a DLL3-positive cancer, the method comprising administering an anti-DLL3 agent to a subject in need thereof, wherein the anti-DLL3 agent is administered according to the following regimen: a) a first dose of from 0.3 mg to 100 mg on day 1 (run-in dose), b) a second dose of from 0.3 mg to 100 mg on day 8 (step dose), and c) one or more subsequent doses of from 0.3 mg to 100 mg starting on day 15 and every two weeks thereafter (target dose), and wherein the second dose and subsequent doses are the same and higher than the first dose.

[0137] In certain embodiments, disclosed herein is a method of treating a DLL3-positive cancer, the method comprising administering an anti-DLL3 agent to a subject in need thereof, wherein the anti-DLL3 agent is administered according to the following regimen: a) a first dose from 0.5 mg to 10 mg on day 1 (run-in dose), b) a second dose from 3 mg to 200 mg on day 8 (step dose), and c) one or more subsequent doses from 3 mg to 200 mg starting on day 15 and every two weeks thereafter (target dose), and wherein the second dose and the subsequent dose are the same and higher than the first dose.

[0138] In certain embodiments of a step dosing regimen, the second dose and subsequent doses are the same and are at least 1.5 times, at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 20 times, at least 25 times, at least 30 times, at least 35 times, at least 40 times, at least 45 times, at least 50 times, at least 55 times, at least 60 times, at least 65 times, at least 70 times, at least 75 times, at least 80 times, at least 85 times, at least 90 times, at least 95 times, at least 100 times, at least 120 times, at least 150 times, or at least 200 times higher than the first dose.

[0139] In certain embodiments of a step dosing regimen, the second dose and subsequent doses are the same and are at least 1.5 times, at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 20 times, at least 25 times, at least 30 times, at least 35 times, at least 40 times, at least 45 times, at least 50 times, at least 55 times, at least 60 times, at least 65 times, at least 70 times, at least 75 times, at least 80 times, at least 85 times, at least 90 times, at least 95 times, or at least 100 times higher than the first dose. And each of the first, second and subsequent doses of the anti-DLL3 agent can be any of the following: from about 0.3 mg to about 100 mg, from about 0.3 mg to about 90 mg, from about 0.3 mg to about 80 mg, from about 0.3 mg to about 70 mg, from about 0.3 mg to about 60 mg, from about 0.3 mg to about 50 mg, from about 0.3 mg to about 40 mg, from about 0.3 mg to about 30 mg, from about 0.3 mg to about 20 mg, from about 0.3 mg to about 10 mg, from about 0.3 mg to about 30 mg, from about 0.3 mg to about 20 mg, from about 0.3 mg to about 10 mg, from about 0.3 mg to about 30 mg, from about 0.3 mg to about 20 mg.from about 1 mg to about 100 mg, from about 1 mg to about 90 mg, from about 1 mg to about 80 mg, from about 1 mg to about 70 mg, from about 1 mg to about 60 mg, from about 1 mg to about 50 mg, from about 1 mg to about 40 mg, from about 1 mg to about 30 mg, from about 1 mg to about 20 mg, from about 1 mg to about 10 mg, from about 1 mg to about 3 mg, from about 3 mg to about 100 mg, from about 3 mg to about 90 mg, from about 3 mg to about 80 mg, from about 3 mg to about 70 mg, from about 3 mg to about 60 mg, from about 3 mg to about 50 mg, from about 1 mg to about 40 mg, from about 1 mg to about 30 mg, from about 1 mg to about 20 mg, from about 1 mg to about 10 mg, from about 3 mg to about 12 mg, from about 3 mg to about 15 mg, from about 10 mg to about 100 mg, from about 10 mg to about From about 90 mg, from about 10 mg to about 80 mg, from about 10 mg to about 70 mg, from about 10 mg to about 60 mg, from about 10 mg to about 50 mg, from about 10 mg to about 40 mg, from about 10 mg to about 30 mg, from about 10 mg to about 20 mg, from about 10 mg to about 15 mg, from about 20 mg to about 100 mg, from about 20 mg to about 90 mg, from about 20 mg to about 80 mg, from about 20 mg to about 70 mg, from about 20 mg to about 60 mg, from about 20 mg to about 50 mg, from about 20 mg to about 40 mg, from about 20 mg to about 30 mg, from about 30 mg to about 100 mg, from about 30 mg to about 90 mg, from about 30 mg to about 80 mg, from about 30 mg to about 70 mg, from about 30 mg to about 60 mg, from about 30 mg to about 50 mg, and from about 30 mg to about 40 mg. .

[0140] In certain embodiments of a step-dosing regimen, a first dose of the anti-DLL3 agent is from about 0.5 mg to about 10 mg, from about 0.5 mg to about 8 mg, from about 0.5 mg to about 6 mg, from about 0.5 mg to about 4 mg, from about 0.5 mg to about 2 mg, or about 1 mg; a second dose and subsequent doses are the same and are at least 3 times, at least 5 times, at least 10 times, at least 20 times, at least 30 times, at least 40 times, at least 50 times, at least 60 times, at least 70 times, at least 80 times, at least 90 times, at least 100 times, at least 120 times, at least 150 times, or at least 200 times higher than the first dose, and each of the second dose and subsequent doses can be any one of the following: from about 3 mg to about 200 mg. g, from about 3 mg to about 100 mg, from about 10 mg to about 180 mg, from about 10 mg to about 150 mg, from about 30 mg to about 200 mg, from about 30 mg to about 180 mg, from about 30 mg to about 150 mg, from about 30 mg to about 120 mg, from about 50 mg to about 200 mg, from about 50 mg to about 180 mg, from about 50 mg to about 150 mg, from about 50 mg to about 120 mg, from about 70 mg to about 200, from about 70 mg to about 180, from about 70 mg to about 150, from about 70 mg to about 120 mg, from about 90 mg to about 200, from about 90 mg to about 180, from about 90 mg to about 120 mg, or about 100 mg.

[0141] In certain embodiments of a step-dosing regimen, the first dose of the anti-DLL3 agent is 1 mg, and the second and subsequent doses are the same and are 3 mg, 10 mg, 30 mg, or 100 mg.

[0142] Disclosed herein, in certain embodiments, are methods of treating a DLL3-positive cancer, the methods comprising administering an anti-DLL3 agent to a subject in need thereof, wherein the anti-DLL3 agent is administered according to a two-step dosing regimen.

[0143] In certain embodiments, disclosed herein is a method of treating a DLL3-positive cancer, the method comprising administering an anti-DLL3 agent to a subject in need thereof, wherein the anti-DLL3 agent is administered according to one of the following two regimens:

[0144] Regimen I: a) a first dose of 0.5 mg to 10 mg on day 1 (run-in dose), b) a second dose of 3 mg to 100 mg on day 4 (step dose), c) a third dose of 3 mg to 200 mg on day 8 (step dose), and d) one or more subsequent doses from 3 mg to 200 mg starting on day 15 and every two weeks thereafter (target dose), wherein the second dose is higher than the first dose, and the third dose and the subsequent dose are the same and higher than the second dose; or

[0145] Regimen II: a) a first dose of 0.5 mg to 10 mg on day 1 (run-in dose), b) a second dose of 3 mg to 100 mg on day 8 (step dose), c) a third dose of 3 mg to 200 mg on day 15 (step dose), and c) one or more subsequent doses of 3 mg to 200 mg starting on day 29 and every two weeks thereafter (target dose), wherein the second dose is higher than the first dose, and the third dose and the subsequent dose are the same and higher than the second dose.

[0146] In certain embodiments of the two-step dosing regimen, the second dose is at least 20 times, at least 30 times, at least 40 times, at least 50 times, at least 60 times, at least 70 times, at least 80 times, or at least 90 times higher than the first dose, and the third dose and the subsequent dose are the same and are at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, or at least 10 times higher than the second dose.

[0147] In certain embodiments of the two-step dosing regimen, the second dose is from about 20 times to about 100 times, from about 20 times to about 90 times, from about 20 times to about 70 times, from about 20 times to about 50 times, from about 25 times to about 50 times, from about 30 times to about 100 times, from about 30 times to about 90 times, from about 30 times to about 70 times, or from about 30 times to about 50 times higher than the first dose, and the third dose and the subsequent dose are the same and are from about 2 times to about 10 times, from about 2 times to about 8 times, from about 2 times to about 6 times, from about 2 times to about 4 times, from about 4 times to about 8 times, or from about 4 times to about 6 times higher than the second dose.

[0148] In certain embodiments of the two-step dosing regimen, the first dose is from about 0.5 mg to about 10 mg, from about 0.5 mg to about 8 mg, from about 0.5 mg to about 6 mg, from about 0.5 mg to about 4 mg, from about 0.5 mg to about 2 mg, or 1 mg, the second dose is from about 3 mg to about 10 mg, from about 10 mg to about 100 mg, from about 10 mg to about 80 mg, from about 10 mg to about 60 mg, from about 20 mg to about 80 mg, from about 20 mg to about 60 mg, from about 25 mg to about 50 mg, from about 30 mg to about 80 mg, from about 30 mg to about 60 mg, from about 30 mg to about 40 mg, from about 40 mg to about 80 mg, or from about 40 mg to about 60 mg, and the third dose and the subsequent dose are the same and range from about 3 mg to about 100 mg, from about 10 mg to about 200 mg, from about 1 From about 10 mg to about 150 mg, from about 10 mg to about 120 mg, from about 30 mg to about 200 mg, from about 30 mg to about 180 mg, from about 30 mg to about 150 mg, from about 30 mg to about 120 mg, from about 50 mg to about 200 mg, from about 50 mg to about 180 mg, from about 50 mg to about 150 mg, from about 70 mg to about 200 mg, from about 70 mg to about 180 mg, from about 70 mg to about 150 mg, from about 70 mg to about 120 mg, from about 90 mg to about 200 mg, from about 90 mg to about 180 mg, from about 90 mg to about 150 mg, from about 90 mg to about 120 mg, from about 100 mg to about 180 mg, from about 100 mg to about 150 mg, from about 100 mg to about 120 mg, or about 100 mg.

[0149] In certain embodiments of the two-step dosing regimen, the first dose is 1 mg, the second dose is from about 25 mg to about 50 mg, and the third dose is the same as the subsequent dose and is 100 mg. In certain embodiments of the two-step dosing regimen, the first dose is 1 mg, the second dose is from about 45 mg to about 70 mg, and the third dose is the same as the subsequent dose and is 100 mg.

[0150] In certain embodiments of the two-step dosing regimen, the first dose on day 1 is 1 mg, the second dose on day 4 is 25 mg or 50 mg, the third dose on day 8 is 100 mg, and starting on day 15 and every two weeks thereafter, one or more subsequent doses are 100 mg.

[0151] In certain embodiments of Regimen I, the target dose is administered on Day 4. In such embodiments, the anti-DLL3 agent is administered according to the following regimen: a) a first dose from 0.5 mg to 10 mg on Day 1 (run-in dose), b) a second dose from 3 mg to 200 mg on Day 4 (step dose, equal to the target dose), c) a third dose from 3 mg to 200 mg on Day 8 (target dose), and d) one or more subsequent doses from 3 mg to 200 mg starting on Day 15 and every two weeks thereafter (target dose), wherein the second dose is higher than the first dose, and the third dose and the subsequent dose are the same, and are the same as the second dose. In certain embodiments, the first dose is from 0.5 mg to 8 mg, from 0.5 mg to 6 mg, from 0.5 mg to 4 mg, from 0.5 mg to 2 mg, or 1 mg, and the second, third, and subsequent doses are each from 3 mg to 100 mg, from 10 mg to 150 mg, from 30 mg to 200 mg, from 30 mg to 100 mg, from 50 mg to 150 mg, from 70 mg to 120 mg, from 90 mg to 120 mg, from 100 mg to 200 mg, or 100 mg. It is believed that such a dosing regimen (target doses administered on days 4, 8, and 15 of the first cycle) is beneficial because it helps to quickly achieve the desired serum levels of the anti-DLL3 agent.

[0152] Disclosed herein, in certain embodiments, are methods of treating a DLL3-positive cancer, the methods comprising administering an anti-DLL3 agent to a subject in need thereof, wherein the anti-DLL3 agent is administered according to a three-step dosing regimen.

[0153] In certain embodiments, disclosed herein is a method for treating DLL3-positive cancer, the method comprising administering an anti-DLL3 agent to a subject in need thereof, wherein the anti-DLL3 agent is administered according to the following regimen: a) a first dose from 0.5 mg to 10 mg on day 1 (run-in dose), b) a second dose from 3 mg to 100 mg on day 4 (step dose), c) a third dose from 3 mg to 100 mg on day 8 (step dose), d) a fourth dose from 3 mg to 200 mg on day 15 (step dose), and e) one or more subsequent doses from 3 mg to 200 mg starting on day 29 and every two weeks thereafter (target dose), and wherein the second dose is higher than the first dose, the third dose is higher than the second dose, and the fourth dose and the subsequent dose are the same and higher than the third dose.

[0154] In certain embodiments of the three-step dosing regimen, the second dose is at least 5 times, at least 10 times, at least 20 times, at least 30 times, at least 40 times, or at least 50 times higher than the first dose, the third dose is at least 0.5 times, at least 1 times, at least 2 times, at least 3 times, or at least 5 times higher than the second dose, and the fourth and subsequent doses are the same and are at least 1 times, at least 2 times, at least 3 times, or at least 5 times higher than the third dose.

[0155] In certain embodiments of the three-step dosing regimen, the second dose is from about 5 to about 70 times, from about 20 to about 70 times, from about 20 to about 60 times, from about 25 to about 50 times higher than the first dose, the third dose is from about 0.5 to about 4 times, from about 0.5 to about 2.5 times, from about 1 to about 4 times, or from about 1 to about 2 times higher than the second dose, and the fourth and subsequent doses are the same and are from about 1 to about 4 times or from about 2 to about 3 times higher than the third dose.

[0156] In certain embodiments of the three-step dosing regimen, the first dose is from about 0.5 mg to about 10 mg, from about 0.5 mg to about 8 mg, from about 0.5 mg to about 6 mg, from about 0.5 mg to about 4 mg, from about 0.5 mg to about 2 mg, or 1 mg, the second dose is from about 3 mg to about 10 mg, from about 10 mg to about 100 mg, from about 10 mg to about 80 mg, from about 10 mg to about 60 mg, from about 10 mg to about 40 mg, from about 20 mg to about 80 mg, from about 20 mg to about 60 mg, from about 20 mg to about 40 mg, or about 25 mg, and the third dose is from about 3 mg to about 10 mg, from about 10 mg to about 100 mg, from about 10 mg to about 80 mg, from about 10 mg to about 60 mg, from about 20 mg to about 100 mg, from about 20 mg to about 80 mg, from about 20 mg to about 60 mg, from about 30 mg to about 100 mg, from about 30 mg to about 30 mg g to about 80 mg, from about 30 mg to about 60 mg, from about 40 mg to about 100 mg, from about 40 mg to about 80 mg, from about 40 mg to about 60 mg, or about 50 mg, and the fourth and subsequent doses are the same and are from about 3 mg to about 10 mg, from about 10 mg to about 200 mg, from about 10 mg to about 150 mg, from about 30 mg to 200 mg, from about 30 mg to about 150 mg, from about 30 mg to about 100 mg, From about 50 mg to about 200 mg, from about 50 mg to about 180 mg, from about 50 mg to about 150 mg, from about 50 mg to about 120 mg, from about 70 mg to about 200 mg, from about 70 mg to about 180 mg, from about 70 mg to about 120 mg, from about 90 mg to about 200 mg, from about 90 mg to about 180 mg, from about 90 mg to about 150 mg, from about 90 mg to about 120 mg, or about 100 mg.

[0157] In certain embodiments of the three-step dosing regimen, the first dose is 1 mg, the second dose is 25 mg, the third dose is 50 mg, and the fourth and subsequent doses are the same and are 100 mg.

[0158] In certain embodiments of the three-step dosing regimen, the target dose is administered on day 8. In such embodiments, the anti-DLL3 agent is administered according to the following regimen: a) a first dose from 0.5 mg to 10 mg on day 1 (run-in dose), b) a second dose from 3 mg to 100 mg on day 4 (step dose), c) a third dose from 3 mg to 200 mg on day 8 (step dose, equal to the target dose), d) a fourth dose from 3 mg to 200 mg on day 15 (target dose), and e) one or more subsequent doses from 3 mg to 200 mg starting on day 29 and every two weeks thereafter (target dose), and wherein the second dose is higher than the first dose, the third dose is higher than the second dose, and the fourth dose and the subsequent dose are the same, and are the same as the third dose. In certain embodiments, the first dose is from 0.5 mg to 8 mg, from 0.5 mg to 6 mg, from 0.5 mg to 4 mg, from 0.5 mg to 2 mg, or 1 mg, the second dose is from 3 mg to 10 mg, from 10 mg to 80 mg, from 10 mg to 60 mg, from 20 mg to 80 mg, from 20 mg to 60 mg, from 20 mg to 40 mg, 25 mg, or 50 mg, and the third, fourth, and subsequent doses are each from 3 mg to 100 mg, from 10 mg to 150 mg, from 30 mg to 200 mg, from 30 mg to 100 mg, from 50 mg to 150 mg, from 70 mg to 120 mg, from 90 mg to 120 mg, 100 mg to 200 mg, or 100 mg. It is believed that such a dosing regimen is beneficial because it helps to quickly achieve the desired serum level of the anti-DLL3 agent.

[0159] The anti-DLL3 agent can be administered by any suitable means, including parenteral, subcutaneous, intraperitoneal, intrapulmonary, intranasal and / or intralesional administration. Parenteral administration includes intramuscular, intravenous, intraarterial, intraperitoneal or subcutaneous administration. In certain embodiments, the anti-DLL3 agent is administered by intravenous (IV) infusion.

[0160] In certain embodiments, the DLL3-positive cancer is small cell lung cancer (SCLC). In certain embodiments, the SCLC is relapsed / refractory SCLC (RR SCLC) or extensive disease SCLC (ED SCLC). In certain embodiments, the subject is a human with SCLC (e.g., RR SCLC or ED SCLC).

[0161] 3.2 Co-administration

[0162] In some embodiments, the compositions and methods of the present invention provide for the use of an anti-DLL3 agent in combination with one or more additional therapeutic agents. In certain embodiments, one or more additional therapeutic agents are agents that alleviate CRS of anti-DLL3 agents (e.g., AMG 757). In certain embodiments, one or more additional therapeutic agents include anti-inflammatory agents, fluids (e.g., saline), anti-IL6 antibodies (e.g., tocilizumab) or anti-TNF agents (e.g., etanercept). In certain embodiments, the methods disclosed herein include administering an anti-DLL3 agent (e.g., AMG 757) in combination with one or more anti-inflammatory agents, fluids (e.g., saline), anti-IL6 antibodies (e.g., tocilizumab) and anti-TNF agents (e.g., etanercept).

[0163] In some embodiments, one or more additional therapeutic agents may be an anti-inflammatory agent (e.g., a prophylactic treatment of CRS). The anti-inflammatory agent may be administered prior to, concurrently with, or after the administration of an anti-DLL3 agent. Exemplary anti-inflammatory agents include acetaminophen, naproxen sodium, ibuprofen, tramadol, aspirin, celecoxib, valdecoxib, indomethacin, or other nonsteroidal anti-inflammatory drugs (NSAIDs). Other anti-inflammatory agents include, for example, beclomethasone, hydroxycortisone, betamethasone, methylprednisolone, budesonide, prednisolone, cortisone, prednisone, dexamethasone, and triamcinolone acetonide or other glucocorticoids. In certain embodiments, the anti-inflammatory agent is a corticosteroid. In certain embodiments, the corticosteroid is dexamethasone. In certain embodiments, the anti-inflammatory agent is acetaminophen. In certain embodiments, an anti-inflammatory agent (e.g., a corticosteroid such as dexamethasone or acetaminophen) is administered prior to the administration of an anti-DLL3 agent. In certain embodiments, dexamethasone is administered intravenously, for example, prior to the cycle 1 dosing of AMG 757. In certain embodiments, dexamethasone is administered orally.

[0164] In some embodiments, one or more additional therapeutic agents that alleviate CRS are fluids (e.g., saline), anti-IL6 antibodies (e.g., tocilizumab), or anti-TNF agents (e.g., etanercept). Each of these agents may be administered prior to, concurrently with, or after the administration of the anti-DLL3 agent. In certain embodiments, saline is administered after the anti-DLL3 agent is administered (e.g., by intravenous administration). In certain embodiments, an anti-IL6 antibody (e.g., tocilizumab) or anti-TNF agent (e.g., etanercept) is administered prior to the administration of the anti-DLL3 agent.

[0165] When steroids (eg, dexamethasone) are used, higher doses of the anti-DLL3 agent may be required. Thus, in some aspects, the invention provides a method of treating SCLC or DLL3-positive cancer, the method comprising administering a steroid (e.g., a corticosteroid (e.g., dexamethasone)) and an anti-DLL3 agent to a subject in need thereof, wherein the anti-DLL3 agent is administered at a dose of from about 0.3 mg to 100 mg once every two weeks (e.g., from about 0.3 mg to about 90 mg, from about 0.3 mg to about 80 mg, from about 0.3 mg to about 70 mg, from about 0.3 mg to about 60 mg, from about 0.3 mg to about 50 mg, from about 0.3 mg to about 40 mg, from about 0.3 mg to about 30 mg, from about 0.3 mg to about 20 mg, from about 0.3 mg to about 10 mg, from about 0.3 mg to about 3 mg, from about 0.3 mg to about 1 mg, from about 1 mg to about 30 mg, from about 1 mg to about 20 mg, from about 3 mg to about 15 mg, or from about 3 mg to about 12 mg, once every two weeks).

[0166] In some aspects, the invention provides a method of treating SCLC or DLL3-positive cancer, the method comprising administering a steroid (e.g., a corticosteroid (e.g., dexamethasone)) and an anti-DLL3 agent to a subject in need thereof, wherein the anti-DLL3 agent is administered once every two weeks at a dose of from about 0.3 mg to 100 mg or from about 3 mg to about 200 mg (e.g., from about 3 mg to about 100 mg, from about 10 mg to about 180 mg, from about 10 mg to about 150 mg, from about 10 mg to about 120 mg, from about 30 mg to about 200 mg, from about 3 ...50 mg, from about 10 mg to about 120 mg, from about 30 mg to about 150 mg, from about 10 mg to about 150 mg, from about 10 mg to about 150 mg, from about 10 mg to about 150 mg, from about 10 mg to about 150 mg, from about From about 30 mg to about 150 mg, from about 30 mg to about 120 mg, from about 50 mg to about 200 mg, from about 50 mg to about 180 mg, from about 50 mg to about 150 mg, from about 50 mg to about 120 mg, from about 70 mg to about 180 mg, from about 70 mg to about 150 mg, from about 70 mg to about 120 mg, from about 90 mg to about 180 mg, from about 90 mg to about 150 mg, from about 90 mg to about 120 mg, or about 3 mg, 10 mg, 30 mg, or 100 mg once every two weeks). In certain embodiments, the anti-DLL3 agent is administered in a 28-day cycle. In certain embodiments, the steroid is administered in the first cycle of administration of the anti-DLL3 agent.

[0167] In some aspects, the invention provides a method of treating SCLC or DLL3-positive cancer, the method comprising administering a steroid (e.g., a corticosteroid (e.g., dexamethasone)) and an anti-DLL3 agent to a subject in need thereof, wherein the anti-DLL3 agent is administered according to the following regimen: (a) a first dose of from 0.3 mg to 100 mg on day 1 (e.g., from about 0.3 mg to about 90 mg, from about 0.3 mg to about 80 mg, from about 0.3 mg to about 70 mg, from about 0.3 mg to about 60 mg, from about 0.3 mg to about 50 mg, from about 0.3 mg to about 40 mg, from about 0.3 mg to about 55 mg, from about 0.3 mg to about 60 mg, from about 0.3 mg to about 70 mg, from about 0.3 mg to about 80 mg, from about 0.3 mg to about 90 mg, from about 0.3 mg to about 100 mg, from about 0.3 mg to about 110 mg, from about 0.3 mg to about 120 mg, from about 0.3 mg to about 130 mg, from about 0.3 mg to about 140 mg, from about 0.3 mg to about 150 mg, from about 0.3 mg to about 160 mg, from about 0.3 mg to about 170 mg, from about 0.3 mg to about 180 mg, from about 0.3 mg to about 190 mg, from about 0.3 mg to about 200 mg, from about 0.3 mg to about 210 mg, from about 0.3 mg to about 220 mg, from about 0.3 mg to about 230 mg, from about 0.3 mg to about 240 mg, from about 0.3 mg to about 250 mg, from about 0. from about 0.3 mg to about 30 mg, from about 0.3 mg to about 20 mg, from about 0.3 mg to about 10 mg, from about 0.3 mg to about 3 mg, from about 0.3 mg to about 1 mg, from about 1 mg to about 30 mg, from about 1 mg to about 20 mg, from about 3 mg to about 15 mg, or from about 3 mg to about 12 mg on day 1), (b) a second dose of from 0.3 mg to 100 mg on day 8 (e.g., from about 0.3 mg to about 90 mg, from about 0.3 mg to about 80 mg, from about 0.3 mg to about 70 mg, from about 0.3 mg to about 60 mg, from about 0.3 mg to about 50 mg g, from about 0.3 mg to about 40 mg, from about 0.3 mg to about 30 mg, from about 0.3 mg to about 20 mg, from about 0.3 mg to about 10 mg, from about 0.3 mg to about 3 mg, from about 0.3 mg to about 1 mg, from about 1 mg to about 30 mg, from about 1 mg to about 20 mg, from about 3 mg to about 15 mg, or from about 3 mg to about 12 mg, on day 8), and (c) one or more subsequent doses of from 0.3 mg to 100 mg (e.g., from about 0.3 mg to about 90 mg, from about 0.3 mg to about 80 mg) starting on day 15 and every two weeks thereafter g, from about 0.3 mg to about 70 mg, from about 0.3 mg to about 60 mg, from about 0.3 mg to about 50 mg, from about 0.3 mg to about 40 mg, from about 0.3 mg to about 30 mg, from about 0.3 mg to about 20 mg, from about 0.3 mg to about 10 mg, from about 0.3 mg to about 3 mg, from about 0.3 mg to about 1 mg, or from about 1 mg to about 30 mg, from about 1 mg to about 20 mg, from about 3 mg to about 15 mg, or from about 3 mg to about 12 mg), and wherein the second dose and the one or more subsequent doses are the same and higher than the first dose.In certain embodiments, the second dose and the one or more subsequent doses are at least 1.5 times, at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 20 times, at least 25 times, at least 30 times, at least 35 times, at least 40 times, at least 45 times, at least 50 times, at least 55 times, at least 60 times, at least 65 times, at least 70 times, at least 75 times, at least 80 times, at least 85 times, at least 90 times, at least 95 times, or at least 100 times higher than the first dose.

[0168] In certain embodiments, an anti-DLL3 agent (e.g., AMG 757) is administered to a subject with a DLL3-positive cancer (e.g., SCLC) in 28-day cycles, wherein the anti-DLL3 agent is administered in the first cycle with an anti-inflammatory agent, a fluid (e.g., saline), an anti-IL6 antibody (e.g., tocilizumab), an anti-TNF agent (e.g., etanercept), or a combination thereof. In certain embodiments, a corticosteroid (e.g., dexamethasone) is further administered in the first cycle with the anti-DLL3 agent and the anti-inflammatory agent, a fluid (e.g., saline), an anti-IL6 antibody (e.g., tocilizumab), an anti-TNF agent (e.g., etanercept), or a combination thereof. In certain embodiments, a corticosteroid (e.g., dexamethasone) is administered by intravenous infusion in the first cycle of AMG 757 administration.

[0169] In certain embodiments, fluid (e.g., saline) is administered by intravenous infusion after administration of the anti-DLL3 agent in the first cycle. In certain embodiments, one liter of saline is administered by intravenous infusion after administration of the anti-DLL3 agent in the first cycle. In certain embodiments, one liter of saline is administered by intravenous infusion after a run-in dose and one or more step doses of the anti-DLL3 agent in the first cycle. In some embodiments, one liter of saline is administered by intravenous infusion over about 4-5 hours after administration of the anti-DLL3 agent.

[0170] In certain embodiments, the anti-inflammatory agent is administered prior to administration of the anti-DLL3 agent in the first cycle. In certain embodiments, the anti-inflammatory agent is a corticosteroid (e.g., dexamethasone) and is administered from about 6 to about 16 hours prior to administration of the anti-DLL3 agent in the first cycle. In certain embodiments, about 8 mg of dexamethasone is administered from about 6 to about 16 hours prior to administration of the anti-DLL3 agent in the first cycle. In certain embodiments, about 8 mg of dexamethasone is administered from about 6 to about 16 hours prior to the run-in dose and one or more step doses of the anti-DLL3 agent in the first cycle. In some embodiments, the dexamethasone is administered orally, and in other embodiments, the dexamethasone is administered intravenously.

[0171] In certain embodiments, the anti-inflammatory agent is acetaminophen and is administered about 1 hour prior to administration of the anti-DLL3 agent in the first cycle. In certain embodiments, about 650 mg of acetaminophen is administered about 1 hour prior to administration of the anti-DLL3 agent in the first cycle. In certain embodiments, about 650 mg of acetaminophen is administered about 1 hour prior to administration of the anti-DLL3 agent in the first cycle. In certain embodiments, about 650 mg of acetaminophen is administered about 1 hour prior to the run-in dose and one or more step doses of the anti-DLL3 agent in the first cycle. In some embodiments, acetaminophen is administered orally.

[0172] In certain embodiments, the anti-IL6 antibody is tocilizumab and is administered prior to administration of the anti-DLL3 agent in the first cycle. In certain embodiments, about 8 mg / kg of tocilizumab is administered to the subject by intravenous infusion about one hour prior to administration of the anti-DLL3 agent in the first cycle. In certain embodiments, about 8 mg / kg of tocilizumab is administered to the subject by intravenous infusion about one hour prior to the run-in dose and one or more step doses of the anti-DLL3 agent in the first cycle.

[0173] In certain embodiments, the anti-TNF agent is etanercept (e.g., ), and in the first cycle, prior to administration of the anti-DLL3 agent. In certain embodiments, in the first cycle, etanercept is administered from about 36 hours to about 60 hours prior to administration of the anti-DLL3 agent. In certain embodiments, in the first cycle, etanercept is administered from about 36 hours to about 60 hours prior to a run-in dose and one or more step doses of an anti-DLL3 agent. In certain embodiments, in the first cycle, etanercept is administered 2 days prior to administration of the anti-DLL3 agent. In certain embodiments, in the first cycle, about 50 mg of etanercept is administered subcutaneously 2 days prior to administration of the anti-DLL3 agent. In certain embodiments, in the first cycle, about 50 mg of etanercept is administered subcutaneously 2 days prior to a run-in dose and one or more step doses of an anti-DLL3 agent, except for the step dose on day 4 in the case of a two-step or three-step dosing regimen.

[0174] Exemplary CRS relief strategies for anti-DLL3 agents (e.g., AMG 757) using anti-inflammatory agents (e.g., dexamethasone or acetaminophen), saline, anti-IL6 antibodies (e.g., tocilizumab), or anti-TNF agents (e.g., etanercept) are shown in the table below. In certain embodiments, in addition to the CRS relief strategies listed below, a corticosteroid (e.g., dexamethasone) is further administered (e.g., by intravenous infusion) during the first cycle of AMG 757 administration (e.g., administered prior to the first cycle of AMG 757 administration).

[0175] Exemplary CRS mitigation strategy (step-by-step dosing, cycle 1 only)

[0176]

[0177] * :Etanercept can be administered within ±12 hours from the scheduled dose in the table above.

[0178] Exemplary CRS Mitigation Strategy (Two-Step Dosing, Option 1, Cycle 1 Only)

[0179]

[0180] * : Etanercept can be administered within ±12 hours of the scheduled dose in the table above. Exemplary CRS relief strategy (two-step dosing, option 2, cycle 1 only)

[0181]

[0182]

[0183] * : Etanercept can be administered within ±12 hours of the scheduled dose in the table above. Exemplary CRS relief strategy (three-step dosing, cycle 1 only)

[0184]

[0185]

[0186] * :Etanercept can be administered within ±12 hours from the scheduled dose in the table above.

[0187] In some aspects, the invention provides a method of treating SCLC, or DLL3-positive cancer, comprising administering to a subject in need thereof an anti-DLL3 agent and an anti-inflammatory agent (e.g., a corticosteroid (e.g., dexamethasone), or acetaminophen), saline, an anti-IL6 antibody (e.g., tocilizumab), or an anti-TNF agent (e.g., etanercept), and wherein the anti-DLL3 agent is administered in a 28-day cycle according to the following one-step schedule: a) from 0.5 mg to 10 mg (e.g., from about 0.5 mg to about 8 mg, from about 0.5 mg to about 6 mg, from about 0.5 mg to about 4 mg, from about 0.5 mg to about 2 mg, or about 1 mg) on ​​day 1 ), b) a first dose (run-in dose) on day 8 from 3 mg to 200 mg (e.g., from about 3 mg to about 100 mg, from about 10 mg to about 180 mg, from about 10 mg to about 150 mg, from about 30 mg to about 200 mg, from about 30 mg to about 180 mg, from about 30 mg to about 150 mg, from about 30 mg to about 120 mg, from about 50 mg to about 200 mg, from about 50 mg to about 180 mg, from about 50 mg to about 150 mg, from about 50 mg to about 120 mg, from about 70 mg to about 200 mg, from about 70 mg to about 180 mg, from about 70 mg to about 150 mg, from about 70 mg to about 1 c) a second dose (step dose) from about 3 mg to 200 mg (e.g., from about 3 mg to about 100 mg, from about 10 mg to about 180 mg, from about 10 mg to about 150 mg, from about 30 mg to about 200 mg, from about 30 mg to about 180 mg, from about 30 mg to about 150 mg, from about 30 mg to about 120 mg, from about 50 mg to about 200 mg, from about 50 mg to about 120 mg), or about 100 mg) starting on day 15 and every two weeks thereafter. from about 50 mg to about 150 mg, from about 50 mg to about 120 mg, from about 70 mg to about 200, from about 70 mg to about 180, from about 70 mg to about 150, from about 70 mg to about 120 mg, from about 90 mg to about 200, from about 90 mg to about 180, from about 90 mg to about 150 mg, from about 90 mg to about 120 mg, or about 100 mg), wherein the second dose and subsequent doses are the same and are higher than the first dose, and wherein the anti-inflammatory agent, saline, anti-IL6 antibody, or anti-TNF agent is administered during the first cycle of administration of the anti-DLL3 agent.In certain embodiments, the second dose and subsequent doses are the same and are at least 3 times, at least 5 times, at least 10 times, at least 20 times, at least 30 times, at least 40 times, at least 50 times, at least 60 times, at least 70 times, at least 80 times, at least 90 times, at least 100 times, at least 120 times, at least 150 times, or at least 200 times greater than the first dose.

[0188] In some embodiments of a one-step regimen, the method comprises administering an anti-DLL3 agent and dexamethasone, acetaminophen, saline, tocilizumab, or etanercept, wherein the first dose of the anti-DLL3 agent is 1 mg, the second and subsequent doses of the anti-DLL3 agent are the same and are 3 mg, 10 mg, 30 mg, or 100 mg, wherein dexamethasone, acetaminophen, saline, tocilizumab, or etanercept is administered in the first cycle of administration of the anti-DLL3 agent. In some embodiments of such a one-step regimen, a corticosteroid is further administered by intravenous infusion in the first cycle of AMG 757 administration.

[0189] In some aspects, the invention provides a method of treating SCLC, or DLL3-positive cancer, comprising administering to a subject in need thereof an anti-DLL3 agent and an anti-inflammatory agent (e.g., a corticosteroid (e.g., dexamethasone), or acetaminophen), saline, an anti-IL6 antibody (e.g., tocilizumab), or an anti-TNF agent (e.g., etanercept), wherein the anti-DLL3 agent is administered in a 28-day cycle according to the following two-step regimen: Regimen I: a) from 0.5 mg to 10 mg (e.g., from about 0.5 mg to about 8 mg, from about 0.5 mg to about 6 mg, from about 0.5 mg to about 4 mg, from about 0.5 mg to about 2 mg, or 1 mg) on ​​Day 1 The invention provides a first dose (run-in dose) of 3 mg to 100 mg (e.g., from about 3 mg to about 10 mg, from about 10 mg to about 80 mg, from about 10 mg to about 60 mg, from about 20 mg to about 80 mg, from about 20 mg to about 60 mg, from about 25 mg to about 50 mg, from about 30 mg to about 80 mg, from about 30 mg to about 60 mg, from about 30 mg to about 40 mg, from about 40 mg to about 80 mg, or from about 40 mg to about 60 mg) on ​​day 4, c) a third dose (step dose) from 3 mg to 200 mg on day 8, and d) a third dose (step dose) from 3 mg to 200 mg starting on day 15 and every two weeks thereafter. one or more subsequent doses (target doses) from 3 mg to 200 mg, and wherein the second dose is higher than the first dose, and the third dose and the subsequent dose are the same and higher than the second dose, and the third dose and the subsequent dose can be any of the following: from about 3 mg to about 100 mg, from about 10 mg to about 180 mg, from about 10 mg to about 150 mg, from about 10 mg to about 120 mg, from about 30 mg to about 200 mg, from about 30 mg to about 180 mg, from about 30 mg to about 150 mg, from about 30 mg to about 120 mg, from about 50 mg to about 200 mg, from about 50 mg to about 150 mg, In some embodiments of the two-step regimen, the anti-inflammatory agent, saline, anti-IL6 antibody, or anti-TNF agent is administered in the first cycle of administration of the anti-DLL3 agent. In some embodiments of such a two-step regimen, the corticosteroid is further administered by intravenous infusion in the first cycle of AMG 757 administration.

[0190] In certain embodiments, the method comprises administering to a subject in need thereof an anti-DLL3 agent (e.g., AMG757) and dexamethasone, acetaminophen, saline, tocilizumab, or etanercept, wherein the anti-DLL3 agent is administered in a 28-day cycle according to the following two-step schedule: a first dose of 1 mg on day 1, a second dose of 25 mg or 50 mg on day 4, a third dose of 100 mg on day 8, and one or more subsequent doses of 100 mg starting on day 15 and every two weeks thereafter, and wherein dexamethasone, acetaminophen, saline, tocilizumab, or etanercept is administered in the first cycle of administration of the anti-DLL3 agent. In certain embodiments, dexamethasone, acetaminophen, saline, tocilizumab, or etanercept is administered on the same day or before one or more run-in doses and one or more step doses of AMG 757 in cycle 1. In certain embodiments, dexamethasone is further administered by intravenous infusion in cycle 1 of AMG 757 administration.

[0191] In certain embodiments of Regimen I, the target dose is administered on Day 4. In such embodiments, the method comprises administering an anti-DLL3 agent and dexamethasone, acetaminophen, saline, tocilizumab, or etanercept to a subject in need thereof, wherein the anti-DLL3 agent is administered in a 28-day cycle according to the following two-step regimen: Regimen I: a) a first dose (run-in dose) from 0.5 mg to 10 mg (e.g., from about 0.5 mg to about 8 mg, from about 0.5 mg to about 6 mg, from about 0.5 mg to about 4 mg, from about 0.5 mg to about 2 mg, or 1 mg) on ​​Day 1, b) a first dose (run-in dose) from 0.5 mg to 10 mg (e.g., from about 0.5 mg to about 8 mg, from about 0.5 mg to about 6 mg, from about 0.5 mg to about 4 mg, from about 0.5 mg to about 2 mg, or 1 mg) on ​​Day 1, c) a second dose (step dose) from 3 mg to 200 mg on day 4, c) a third dose (target dose) from 3 mg to 200 mg on day 8, and d) one or more subsequent doses (target doses) from 3 mg to 200 mg starting on day 15 and every two weeks thereafter, wherein the second dose is higher than the first dose, and the third dose and the subsequent dose are the same and the same as the second dose, and the second dose, the third dose and the subsequent dose can be any of the following: from about 3 mg to 200 mg on day 8, and the third dose and the subsequent dose can be any of the following: from about 3 mg to 200 mg on day 15, and ... g to about 100 mg, from about 10 mg to about 180 mg, from about 10 mg to about 150 mg, from about 10 mg to about 120 mg, from about 30 mg to about 200 mg, from about 30 mg to about 180 mg, from about 30 mg to about 150 mg, from about 30 mg to about 120 mg, from about 50 mg to about 200 mg, from about 50 mg to about 180 mg, from about 50 mg to about 150 mg, from about 70 mg to about 200 mg, from about 70 mg to about 180 mg, from about 70 In some embodiments, the dosage of dexamethasone, acetaminophen, saline, tocilizumab, or etanercept is from about 100 mg to about 150 mg, from about 70 mg to about 120 mg, from about 90 mg to about 200 mg, from about 90 mg to about 180 mg, from about 90 mg to about 150 mg, from about 90 mg to about 120 mg, from about 100 mg to about 180 mg, from about 100 mg to about 150 mg, from about 100 mg to about 120 mg, or about 100 mg, and wherein dexamethasone, acetaminophen, saline, tocilizumab, or etanercept is administered in the first cycle of administration of the anti-DLL3 agent. In certain embodiments, dexamethasone, acetaminophen, saline, or tocilizumab is administered on days 1 and 4, or on days 1, 4, and 8 of AMG 757 administration in cycle 1. In certain embodiments, etanercept is administered two days prior to days 1 and 8 of AMG 757 administration in cycle 1. In certain embodiments, etanercept is administered two days prior to day 1 of AMG 757 administration in cycle 1. In certain embodiments, the method further comprises administering dexamethasone by intravenous infusion in cycle 1 of AMG 757 administration.

[0192] In some aspects, the invention provides a method of treating SCLC, or DLL3-positive cancer, comprising administering to a subject in need thereof an anti-DLL3 agent and an anti-inflammatory agent (e.g., a corticosteroid (e.g., dexamethasone), or acetaminophen), saline, an anti-IL6 antibody (e.g., tocilizumab), or an anti-TNF agent (e.g., etanercept), wherein the anti-DLL3 agent is administered in a 28-day cycle according to the following two-step regimen: Regimen II: a) from 0.5 mg to 10 mg on Day 1 (e.g., from about 0.5 mg to about 8 mg, from about 0.5 mg to about 6 mg, from about 0.5 mg to about 4 mg, from about 0.5 mg to about 2 mg, or 1 mg ), b) a first dose (run-in dose) on day 8 from 3 mg to 100 mg (e.g., from about 3 mg to about 10 mg, from about 10 mg to about 80 mg, from about 10 mg to about 60 mg, from about 20 mg to about 80 mg, from about 20 mg to about 60 mg, from about 25 mg to about 50 mg, from about 30 mg to about 80 mg, from about 30 mg to about 60 mg, from about 30 mg to about 40 mg, from about 40 mg to about 80 mg, or from about 40 mg to about 60 mg), c) a third dose (step dose) from 3 mg to 200 mg on day 15, and c) starting on day 29 and every two days thereafter One or more subsequent doses (target dose) from 3 mg to 200 mg once a week, and wherein the second dose is higher than the first dose, and the third dose and the subsequent dose are the same and higher than the second dose, and the third dose and the subsequent dose can be any of the following: from about 3 mg to about 100 mg, from about 10 mg to about 180 mg, from about 10 mg to about 150 mg, from about 10 mg to about 120 mg, from about 30 mg to about 200 mg, from about 30 mg to about 180 mg, from about 30 mg to about 150 mg, from about 30 mg to about 120 mg, from about 50 mg to about 200 mg, from about 5 ... In some embodiments of the two-step regimen, the anti-inflammatory agent, saline, anti-IL6 antibody, or anti-TNF agent is administered in the first cycle of administration of the anti-DLL3 agent. In some embodiments of such a two-step regimen, the corticosteroid is further administered by intravenous infusion in the first cycle of AMG 757 administration.

[0193] In some embodiments of the two-step dosing regimen, the method comprises administering to the subject an anti-DLL3 agent and dexamethasone, acetaminophen, saline, tocilizumab, or etanercept, wherein the first dose of the anti-DLL3 agent is 1 mg, the second dose of the anti-DLL3 agent is from about 25 mg to about 50 mg, and the third and subsequent doses of the anti-DLL3 agent are the same and are 100 mg, and wherein the dexamethasone, acetaminophen, saline, tocilizumab, or etanercept is administered in the first cycle of administration of the anti-DLL3 agent.

[0194] In some embodiments of the two-step dosing regimen, the method comprises administering to the subject an anti-DLL3 agent and dexamethasone, acetaminophen, saline, tocilizumab, or etanercept, wherein the first dose of the anti-DLL3 agent is 1 mg, the second dose of the anti-DLL3 agent is from about 45 mg to about 70 mg, and the third and subsequent doses of the anti-DLL3 agent are the same and are 100 mg, and wherein the dexamethasone, acetaminophen, saline, tocilizumab, or etanercept is administered in the first cycle of administration of the anti-DLL3 agent.

[0195] In certain embodiments of the two-step dosing regimen, the method comprises administering to the subject an anti-DLL3 agent and dexamethasone, acetaminophen, saline, tocilizumab, or etanercept, wherein a first dose of the anti-DLL3 agent on day 1 is 1 mg, a second dose of the anti-DLL3 agent on day 4 is 25 mg or 50 mg, a third dose on day 8 is 100 mg, and starting on day 15 and once every two weeks thereafter, subsequent doses of the anti-DLL3 agent are 100 mg, and wherein dexamethasone, acetaminophen, saline, tocilizumab, or etanercept is administered in the first cycle of administration of the anti-DLL3 agent.

[0196] In some aspects, the invention provides a method of treating SCLC, or DLL3-positive cancer, comprising administering to a subject in need thereof an anti-DLL3 agent and an anti-inflammatory agent (e.g., a corticosteroid (e.g., dexamethasone), or acetaminophen), saline, an anti-IL6 antibody (e.g., tocilizumab), or an anti-TNF agent (e.g., etanercept), and wherein the anti-DLL3 agent is administered in a 28-day cycle according to the following three-step regimen: a) a first dose (run-in dose) from 0.5 mg to 10 mg (e.g., from about 0.5 mg to about 8 mg, from about 0.5 mg to about 6 mg, from about 0.5 mg to about 4 mg, from about 0.5 mg to about 2 mg, or 1 mg) on ​​day 1, b) a first dose (run-in dose) from 3 mg to 10 mg on day 4, c) a first dose (run-in dose) from 0.5 mg to 10 mg on day 4, e) a first dose (run-in dose) from 0.5 mg to 10 mg on day 1, and f) a first dose (run-in dose) from 0.5 mg to 10 mg on day 4. g to 100 mg (e.g., from about 3 mg to about 10 mg, from about 10 mg to about 100 mg, from about 10 mg to about 80 mg, from about 10 mg to about 60 mg, from about 10 mg to about 40 mg, from about 20 mg to about 80 mg, from about 20 mg to about 60 mg, from about 20 mg to about 40 mg, or about 25 mg) a second dose (step dose) on day 8, c) 3 mg to 100 mg (e.g., from about 3 mg to about 10 mg, from about 10 mg to about 100 mg, from about 10 mg to about 80 mg, from about 10 mg to about 60 mg, from about 10 mg to about 40 mg, from about 20 mg to about 80 mg, from about 20 mg to about 60 mg, from about 20 mg to about 40 mg, or about 25 mg) a second dose (step dose) on day 8, from about 100 mg, from about 30 mg to about 80 mg, from about 30 mg to about 60 mg, from about 40 mg to about 100 mg, from about 40 mg to about 80 mg, from about 40 mg to about 60 mg, or about 50 mg) on ​​day 15 (step dose), d) a fourth dose from 3 mg to 200 mg on day 15 (step dose), and e) one or more subsequent doses (target dose) from 3 mg to 200 mg starting on day 29 and every two weeks thereafter, and wherein the second dose is higher than the first dose, the third dose is higher than the second dose, and the fourth dose and the subsequent doses are the same and higher than the third dose, and the fourth and the subsequent doses can be the following: Any of: from about 3 mg to about 100 mg, from about 10 mg to about 150 mg, from about 30 mg to 200 mg, from about 50 mg to about 200 mg, from about 50 mg to about 180 mg, from about 50 mg to about 150 mg, from about 50 mg to about 120 mg, from about 70 mg to about 200 mg, from about 70 mg to about 180 mg, from about 70 mg to about 120 mg, from about 90 mg to about 200 mg, from about 90 mg to about 180 mg, from about 90 mg to about 150 mg, from about 90 mg to about 120 mg, or about 100 mg, and wherein the anti-inflammatory agent, saline, anti-IL6 antibody, or anti-TNF agent is administered in the first cycle of administration of the anti-DLL3 agent.In some embodiments of such a three-step regimen, corticosteroids are further administered by intravenous infusion during cycle 1 of AMG 757 administration.

[0197] In some embodiments of the three-step dosing regimen, the method comprises administering to the subject an anti-DLL3 agent and dexamethasone, acetaminophen, saline, tocilizumab, or etanercept, wherein the first dose of the anti-DLL3 agent is 1 mg, the second dose of the anti-DLL3 agent is 25 mg, the third dose of the anti-DLL3 agent is 50 mg and the fourth and subsequent doses of the anti-DLL3 agent are the same and are 100 mg, and wherein the dexamethasone, acetaminophen, saline, tocilizumab, or etanercept is administered in the first cycle of administration of the anti-DLL3 agent.

[0198] In certain embodiments of the three-step dosing regimen, the target dose is administered on day 8. In such embodiments, the method comprises administering to a subject in need thereof an anti-DLL3 agent and dexamethasone, acetaminophen, saline, tocilizumab, or etanercept, wherein the anti-DLL3 agent is administered in a 28-day cycle according to the following three-step regimen: a) a first dose (run-in dose) from 0.5 mg to 10 mg (e.g., from about 0.5 mg to about 8 mg, from about 0.5 mg to about 6 mg, from about 0.5 mg to about 4 mg, from about 0.5 mg to about 2 mg, or 1 mg) on ​​day 1, b) a second dose (step dose) from 3 mg to 100 mg (e.g., from 3 mg to 10 mg, from 10 mg to 80 mg, from 10 mg to 60 mg, from 20 mg to 80 mg, from 20 mg to 60 mg, from 20 mg to 40 mg, 25 mg, or 50 mg) on ​​day 4, c) a third dose (step dose) from 3 mg to 200 mg on day 8, d) a fourth dose (target dose) from 3 mg to 200 mg on day 15, and e) one or more subsequent doses (target doses) from 3 mg to 200 mg starting on day 29 and every two weeks thereafter, and wherein the second dose is higher than the first dose, the third dose, the fourth dose, and the subsequent doses are the same and higher than the second dose, and the third dose, the fourth dose, and the subsequent doses can be any of: from 3 mg to 100 mg, from 10 mg to 150 mg, from 30 mg to 200 mg, from 30 mg to 100 mg, from 50 mg to 150 mg, from 70 mg to 120 mg, from 90 mg to 120 mg, 100 mg to 200 mg, or 100 mg, and wherein dexamethasone, acetaminophen, saline, tocilizumab, or etanercept is administered in the first cycle of administration of the anti-DLL3 agent. In certain embodiments, dexamethasone, acetaminophen, saline, or tocilizumab is administered on days 1, 4, and 8, or on days 1, 4, 8, and 15 of AMG 757 administration in cycle 1. In certain embodiments, etanercept is administered two days prior to days 1, 8, and 15 of AMG 757 administration in cycle 1. In certain embodiments, etanercept is administered two days prior to days 1 and 8 of AMG 757 administration in cycle 1. In some embodiments of such a three-step regimen, corticosteroids are further administered by intravenous infusion in cycle 1 of AMG 757 administration.

[0199] In certain embodiments of any of the step-dosing regimens, saline (e.g., about 1 liter) is administered by intravenous infusion following administration of the anti-DLL3 agent in the first cycle. In certain embodiments, one liter of saline is administered by intravenous infusion following the run-in dose and one or more step doses of the anti-DLL3 agent in the first cycle. In some embodiments, one liter of saline is administered by intravenous infusion over about 4-5 hours following administration of the anti-DLL3 agent.

[0200] In certain embodiments of any of the stepped dosing regimens, the anti-inflammatory agent is a corticosteroid (e.g., dexamethasone) and is administered about 6 to about 16 hours prior to administration of the anti-DLL3 agent in the first cycle. In certain embodiments, about 8 mg of dexamethasone is administered from about 6 to about 16 hours prior to administration of the anti-DLL3 agent in the first cycle. In certain embodiments, 8 mg of dexamethasone is administered about 6 to about 16 hours prior to a run-in dose and one or more stepped doses of the anti-DLL3 agent in the first cycle. In some embodiments, dexamethasone is administered orally.

[0201] In certain embodiments of any of the stepped dosing regimens, the anti-inflammatory agent is acetaminophen and is administered about 1 hour prior to administration of the anti-DLL3 agent in the first cycle. In certain embodiments, about 650 mg of acetaminophen is administered about 1 hour prior to administration of the anti-DLL3 agent in the first cycle. In certain embodiments, about 650 mg of acetaminophen is administered about 1 hour prior to administration of the anti-DLL3 agent in the first cycle. In certain embodiments, about 650 mg of acetaminophen is administered about 1 hour prior to the run-in dose and one or more stepped doses of the anti-DLL3 agent in the first cycle. In some embodiments, the acetaminophen is administered orally.

[0202] In certain embodiments of any of the stepped dosing regimens, the anti-IL6 antibody tocilizumab is administered prior to administration of the anti-DLL3 agent in the first cycle. In certain embodiments, in the first cycle, about 8 mg / kg of tocilizumab is administered to the subject by intravenous infusion about one hour prior to administration of the anti-DLL3 agent. In certain embodiments, in the first cycle, about 8 mg / kg of tocilizumab is administered to the subject by intravenous infusion about one hour prior to the run-in dose and one or more stepped doses of the anti-DLL3 agent.

[0203] In certain embodiments of any of the step-dosing regimens, the anti-TNF agent etanercept (e.g., ). In certain embodiments, etanercept is administered from about 36 hours to about 60 hours prior to administration of the anti-DLL3 agent in the first cycle. In certain embodiments, etanercept is administered from about 36 hours to about 60 hours prior to the run-in dose and one or more step doses of the anti-DLL3 agent in the first cycle, except for the step dose on day 4 in the case of a two-step or three-step dosing regimen. In certain embodiments, about 50 mg of etanercept is administered 2 days prior to administration of the anti-DLL3 agent in the first cycle. In certain embodiments, about 50 mg of etanercept is administered subcutaneously 2 days prior to the run-in dose and one or more step doses of the anti-DLL3 agent in the first cycle, except for the step dose on day 4 in the case of a two-step or three-step dosing regimen.

[0204] In certain embodiments of any of the step dosing regimens, the subject is a human.

[0205] 4. Products

[0206] Disclosed herein are articles of manufacture comprising: (a) a container comprising an anti-DLL3 agent; and (b) a package insert of instructions for treating a DLL3-positive cancer (or treating SCLC) in a subject, wherein the instructions specify a dose of from about 0.3 mg to about 100 mg, from about 3 mg to about 200 mg, or 100 mg (or any dose range disclosed herein) of the anti-DLL3 agent to be administered to the subject once every two weeks (e.g., on days 1 and 15 of a 28-day cycle). The instructions may also specify administration of the anti-DLL3 agent according to the following schedule: a) a first dose of from 0.3 mg to 100 mg (or any dose range disclosed herein) on day 1, b) a second dose of from 0.3 mg to 100 mg (or any dose range disclosed herein) on day 8, and c) one or more subsequent doses of from 0.3 mg to 100 mg (or any dose range disclosed herein) starting on day 15 and once every two weeks thereafter, and wherein the second dose and the one or more subsequent doses are the same and higher than the first dose.

[0207] The instructions may also specify that the anti-DLL3 agent is administered according to the following regimen: a) a first dose from 0.5 mg to 10 mg (or any dose range disclosed herein) on day 1, b) a second dose from 3 mg to 200 mg (or any dose range disclosed herein) on day 8, and c) one or more subsequent doses from 3 mg to 200 mg (or any dose range disclosed herein) starting on day 15 and every two weeks thereafter, and wherein the second dose and the one or more subsequent doses are the same and higher than the first dose.

[0208] In certain embodiments, the instructions specify administration of the anti-DLL3 agent according to one of the following two schedules: Schedule I: a) a first dose of 0.5 mg to 10 mg (or any dose range disclosed herein) on day 1 (run-in dose), b) a second dose of 3 mg to 100 mg (or any dose range disclosed herein) on day 4 (step dose), c) a third dose of 3 mg to 200 mg (or any dose range disclosed herein) on day 8 (step dose), and d) one or more subsequent doses (target dose) from 3 mg to 200 mg (or any dose range disclosed herein) starting on day 15 and every two weeks thereafter, and wherein the second dose is higher than the first dose, and the third dose and the target dose are higher than the first dose. The subsequent doses are the same and higher than the second dose; or regimen II: a) a first dose (run-in dose) from 0.5 mg to 10 mg (or any dose range disclosed herein) on day 1, b) a second dose (step dose) from 3 mg to 100 mg (or any dose range disclosed herein) on day 8, c) a third dose (step dose) from 3 mg to 200 mg (or any dose range disclosed herein) on day 15, and c) one or more subsequent doses (target dose) from 3 mg to 200 mg (or any dose range disclosed herein) starting from day 29 and once every two weeks thereafter, and wherein the second dose is higher than the first dose, the third dose and the subsequent dose are the same and higher than the second dose.

[0209] In certain embodiments, the instructions specify administration of the anti-DLL3 agent according to the following schedule: a) a first dose (run-in dose) from 0.5 mg to 10 mg (or any dose range disclosed herein) on day 1, b) a second dose (step dose) from 3 mg to 200 mg (or any dose range disclosed herein) on day 4, c) a third dose (step dose) from 3 mg to 200 mg (or any dose range disclosed herein) on day 8, and d) one or more subsequent doses (target dose) from 3 mg to 200 mg (or any dose range disclosed herein) starting on day 15 and every two weeks thereafter, and wherein the second dose is higher than the first dose, and the third dose and the subsequent dose are the same, and are the same as the second dose.

[0210] In certain embodiments, the instructions specify administration of the anti-DLL3 agent according to the following schedule: a) a first dose (run-in dose) from 0.5 mg to 10 mg (or any dose range disclosed herein) on day 1, b) a second dose (step dose) from 3 mg to 100 mg (or any dose range disclosed herein) on day 4, c) a third dose (step dose) from 3 mg to 100 mg (or any dose range disclosed herein) on day 8, d) a fourth dose (step dose) from 3 mg to 200 mg (or any dose range disclosed herein) on day 15, and e) one or more subsequent doses (target dose) from 3 mg to 200 mg (or any dose range disclosed herein) starting on day 29 and every two weeks thereafter, wherein the second dose is higher than the first dose, the third dose is higher than the second dose, and the fourth dose and the subsequent dose are the same and higher than the third dose.

[0211] In certain embodiments, the instructions specify administration of the anti-DLL3 agent according to the following schedule: a) a first dose (run-in dose) from 0.5 mg to 10 mg (or any dose range disclosed herein) on day 1, b) a second dose (step dose) from 3 mg to 100 mg (or any dose range disclosed herein) on day 4, c) a third dose (step dose) from 3 mg to 200 mg (or any dose range disclosed herein) on day 8, d) a fourth dose (step dose) from 3 mg to 200 mg (or any dose range disclosed herein) on day 15, and e) one or more subsequent doses (target dose) from 3 mg to 200 mg (or any dose range disclosed herein) starting on day 29 and every two weeks thereafter, wherein the second dose is higher than the first dose, the third dose is higher than the second dose, and the fourth dose and the subsequent dose are the same, and are the same as the third dose.

[0212] In certain embodiments, the instructions specify that an anti-inflammatory agent (e.g., a corticosteroid (e.g., dexamethasone), or acetaminophen), saline, an anti-IL6 antibody (e.g., tocilizumab), or an anti-TNF agent (e.g., etanercept) is also administered during the first cycle of administration of the anti-DLL3 agent. In certain embodiments, the instructions specify that dexamethasone is further administered during the first cycle of administration of the anti-DLL3 agent (e.g., administered intravenously prior to the cycle of dosing of the anti-DLL3 agent).

[0213] Example

[0214] Example 1 Prediction of human AMG 757 pharmacokinetics and first-in-human dose selection

[0215] The human PK parameters of AMG 757 were predicted using allometric scaling of PK parameters obtained from studies in cynomolgus monkeys over a dose range of 12 to 4500 μg / kg. A two-compartment model with linear elimination was used to characterize the pharmacokinetics of AMG 757 from the pooled cynomolgus monkey data that excluded data from animals that were positive for anti-drug antibodies after administration of the first dose. The model employed linear clearance (CL), volume of distribution in the central compartment (V C ), distribution clearance (CL D ) and the peripheral volume of distribution (V T ) parameterization. Allometry was used to predict human AMG 757 PK (using exponents 0.75 and 1 for clearance and volume parameters, respectively). The body weights of humans and cynomolgus monkeys were assumed to be 60 kg and 3 kg, respectively. The derived monkey and predicted human AMG 757 PK parameters are provided in Table 1.

[0216] Table 1 Pharmacokinetic parameters in cynomolgus monkeys and predicted humans for human AMG 757 exposure prediction

[0217]

[0218] CL = systemic clearance; CL D = distribution clearance; t 1 / 2 = terminal half-life; V C = the central volume of the distribution; V T = peripheral volume of distribution

[0219] a Assume the monkey weighs 3kg

[0220] b Assume that the person weighs 60 kg

[0221] In conjunction with these PK predictions, the FIH starting dose was selected based on the lowest anticipated biological effect level (MABEL) identified in vitro. This concentration was determined by assessing the most sensitive marker of AMG 757 activity in the most sensitive DLL3-expressing cell line (Study 123564). In conjunction with animal exposures in GLP toxicology studies, a 50-mg / mL AUC-based toxicity test was used. tau (168 hours in cynomolgus monkeys and 336 hours in humans in GLP toxicology studies) and the recommended dose of C in FIH studies max The exposure margin is calculated using the standard ratio calculation.

[0222] The FIH study evaluated the safety, tolerability, and pharmacokinetics of AMG 757 in patients with small cell lung cancer. The predicted human PK parameters described above were used to simulate the predicted exposure at the proposed FIH dose ( Figure 1The FIH study of AMG757 doses of 0.003 mg, 0.01 mg, 0.03 mg, 0.1 mg, 0.3 mg, 1 mg, 3 mg, 10 mg, 30 mg, 100 mg, and higher doses were administered as a short-term intravenous infusion (approximately 1 hour) every two weeks in patients with small cell lung cancer (SCLC) (Study 20160323). This regimen ensured that in vitro EC values ​​exceeding AMG757-mediated cell killing (5.2 ng / mL) were achieved in the lung over the course of multiple treatment cycles of AMG757. 90 adequate exposure while minimizing the peak-to-valley ratio.

[0223] The starting dose for the FIH study was selected based on the in vitro MABEL. Briefly, AMG 757-mediated cell killing of SHP-77 tumor cells and induction of EC activation (de novo expression of CD69 and CD25) in human peripheral blood mononuclear cells (PBMCs) were compared. 50 The MABEL (mean EC 50 ) was 0.61 ng / mL (5.8 pM; Study 123564).

[0224] Prior safe implementation of this strategy supports the use of in vitro EC 50 As a basis for MABEL and FIH starting doses to identify previously unsuccessful studies in clinical development The maximum recommended starting dose of the molecule.

[0225] Example 2 Prediction of the Minimum Effective Exposure of AMG 757 in Humans

[0226] The effective exposure of AMG 757 was predicted based on in vitro data generated in Study 122717, which evaluated the in vitro pharmacology of AMG 757. The half-maximal effect (EC 50 ) and 90% of maximum effect (EC 90 ) concentration to assess the concentration range in which therapeutic effects can be expected.

[0227] The lung was used as a representative site of action for AMG 757 and was assumed to achieve an exposure of approximately 1% of free serum exposure (Vugmeyser et al., J Pharm Sci. 99:1028-1045 (2010)). Based on this assumption, the ECs that provide AMG 757-mediated cell killing in SHP-77 cells were 50The dose of AMG757 with the trough coverage was considered the lowest effective. The mean EC for cell killing in SHP-77 cells (assuming 1% lung exposure) over the entire treatment cycle was 90 Trough coverage at 10 mg IV every 2 weeks predicts early signs of efficacy ( Figure 2 ).

[0228] Example 3 Phase 1 study evaluating the safety, tolerability, and pharmacokinetics of AMG 757 in subjects with SCLC

[0229] background

[0230] Small cell lung cancer (SCLC), which accounts for 10%-15% of lung cancers (Rudin et al., J Clin Oncol. 33:4106-4111 (2015)), is an aggressive subtype of lung cancer with neuroendocrine differentiation and is closely associated with smoking (Koinis et al., Transl Lung Cancer Res. 5:39-50 (2016)). It shows a distinct natural history characterized by high growth fractions, rapid doubling times, and early establishment of extensive metastatic lesions (Gustafsson et al., Cancer. 113:5-21 (2008)). Although 30% of patients present with disease confined to one hemithorax [limited disease (LD)], the majority of cases have disease that is not covered by one field of radiotherapy [extended disease (ED)]. SCLC is extremely sensitive to first-line chemotherapy (approximately 60%-70% efficacy) and radiation therapy, which is in stark contrast to subsequent resistance to second-line and subsequent therapies after disease recurrence (Byers et al., Cancer. [Cancer] 121: 664672 (2015)). ED patients develop drug resistance and die from the disease in a median time of 10 to 12 months from diagnosis (Rudin et al., 2015). For patients with ED SCLC, first-line treatment is platinum-based chemotherapy. In the United States, most patients receive platinum-etoposide (EP) chemotherapy (carboplatin or cisplatin), and some patients receive platinum-irinotecan as an alternative, especially outside the United States. In March 2019, the U.S. Food and Drug Administration (US FDA) approved atezolizumab for use with carboplatin and etoposide as a first-line treatment for adult patients with ED-SCLC ( U.S. Prescribing Information [USPI], 2019). After relapse, topotecan is the only second-line drug approved by the US FDA. However, despite this, topotecan has produced disappointing response rates in this setting (Byers et al., Cancer. 121:664-672 (2015)).

[0231] AMG 757 is a half-life extended (HLE) AMG 757 is a molecule that targets DLL3 as a tumor-specific antigen and the T-cell receptor-associated complex cluster of differentiation antigen 3 (CD3) on T cells. AMG 757 is developed for the treatment of SCLC and is a potent molecule that works by forming an immunological synapse between CD3-positive T cells and cancer cells expressing the DLL3 protein. The resulting proximity triggers redirected lysis of DLL3-positive target cells by T cells. AMG 757 monotherapy significantly inhibited the growth of subcutaneously implanted DLL3-expressing human melanoma WM266-4 cells and induced regression of orthotopic SHP-77-Luc lung cancers.

[0232] Study Design

[0233] Study 20160323 is an open-label, ascending, multiple-dose Phase 1 study evaluating AMG 757 administered as a short-term intravenous (IV) infusion every 2 weeks (e.g., with or without step dosing on day 8) in subjects with small cell lung cancer. There are two indications in this study: A: relapsed / refractory small cell lung cancer (RR-SCLC) and B: extensive disease SCLC (ED-SCLC).

[0234] Due to its known mechanism of action, subjects are at increased risk of CRS during the initiation of AMG 757 treatment.

[0235] This study consists of three parts:

[0236] (1) Part A: AMG 757 was evaluated in subjects with relapsed / refractory small cell lung cancer (RR SCLC). Part A consisted of two phases: (A1) a dose-finding phase to determine the maximum tolerated dose (MTD) or recommended phase 2 dose (RP2D) of AMG 757, and (A2) a dose-expansion phase to confirm the safety and tolerability of the selected dose.

[0237] (2) Part B: Evaluate AMG 757 in patients with extensive-disease SCLC (ED SCLC). Once the MTD or RP2D is identified in Part A, Part B will begin.

[0238] (3) Part C (Evaluation of additional CRS relief strategies): AMG 757 monotherapy for subjects with RR SCLC that has progressed after at least 1 platinum-based regimen. To mitigate the risk of CRS, one or more of the following preventive measures may be implemented (during cycle 1 only): intravenous rehydration, additional corticosteroid prophylaxis with oral dexamethasone, administration of tocilizumab prophylaxis, etanercept prophylaxis, or acetaminophen prophylaxis.

[0239] The primary objectives of both Part A and Part B of this study are to evaluate the safety and tolerability of AMG 757 and to determine the MTD or RP2D of AMG 757. The secondary objectives of Part A and Part B of this study are to characterize the pharmacokinetics (PK) of AMG 757 and to preliminarily evaluate the anti-tumor activity of AMG 757.

[0240] Part A1: Dose-finding phase

[0241] AMG 757 is administered as a short intravenous infusion (approximately 60 minutes). The pre-specified doses used in the dose escalation (the planned dose levels for the study) are: 0.003 mg (Group 1), 0.01 mg (Group 2), 0.03 mg (Group 3), 0.1 mg (Group 4), 0.3 mg (Group 5), 1 mg (Group 6), 3 mg (Group 7), 10 mg (Group 8), 30 mg (Group 9), and 100 mg (Group 10), administered once every two weeks. The dosing regimen in the first cycle can be adjusted to include one or more step doses, as described below. If a convincing clinical response is observed during the incremental phase, further dose escalation can be stopped. Alternative dose levels or one or more dosing regimens of AMG 757 can be explored based on emerging pharmacokinetics (PK), pharmacodynamics (PD) and safety data. If the MTD is not reached at the planned dose group level (Groups 1-10), and safety and PK / PD data support is obtained, a higher dose can be explored.

[0242] Step-dosing: After the initial infusion of AMG 757, subjects may experience a first dose effect (e.g., cytokine release syndrome with associated manifestations and any other potentially evolving and unknown first dose effects). It is believed that the optimal MTD may require a step-dosing approach (e.g., initial dose on day 1 and step dose on day 8). Two MTDs can be estimated, one for the initial dose (MTD1) and one for subsequent doses (MTD2).

[0243] At any time during the study, when a subject experiences a first dose effect (such as a CRS event of any grade) for the first time, the safety data needs to be reviewed to determine the appropriate dose to implement as the initial dose (MTD1) that does not exceed the dose at which Grade 2 or higher CRS is observed. These doses and dosing schedules are guided by modeling and simulations of emerging clinical data (such as pharmacokinetics, safety data, etc.) to ensure that the systemic exposure of AMG 757 does not exceed the dose associated with the observed first dose effect. The increased dosing frequency and possible drug accumulation of the step-dosing schedule need to be considered.

[0244] For all subjects enrolled in the follow-up cohort throughout the study, dose escalation continues at a constant dose for the first dose of the first cycle of AMG 757 (MTD1), and only step doses will be escalated according to the pre-specified doses in the dose escalation to determine MTD2. Figure 3 An example of stepped dosing is shown in .

[0245] The step-dose regimens are summarized below. According to DLRT recommendations based on emerging safety data, the dosage regimen may be adapted to include one or more of the following measures.

[0246] • Single-step dosing involves a run-in dose on Day 1, followed by a step dose (equal to the target dose) on Day 8 and the target dose on Day 15 and then Q2W.

[0247] Two-step dosing (option 1) involves a run-in dose on day 1, followed by a step dose on day 4, a step dose on day 8 (equal to the target dose), and a target dose on day 15, then Q2W.

[0248] Two-step dosing (Option 2) involves a run-in dose on Day 1, followed by a step dose on Day 8, a step dose on Day 15 (equal to the target dose), and the target dose on Day 1 of Cycle 2, then Q2W.

[0249] Three-step dosing involves a run-in dose on Day 1, followed by a step dose on Day 4, a step dose on Day 8, a step dose on Day 15 (equal to the target dose), and a target dose on Day 1 of Cycle 2, then Q2W.

[0250] Part A2: Dose Expansion Phase: Once the MTD or RP2D is selected based on the dose-finding phase (Part A1), Part A2 begins.

[0251] Part B: Part B will begin once the preliminary MTD or RP2D in Indication A (Part A1) is established.

[0252] Part C: Following the DLRT recommendations based on emerging safety data, one or more additional CRS mitigation strategies as outlined below may be evaluated. Part C will begin while Part A1 is ongoing.

[0253] Immediately after the run-in dose and one or more step doses in cycle 1, give 1 L of normal saline intravenously over 4-5 hours for prophylaxis

[0254] Additional corticosteroid prophylaxis with 8 mg of dexamethasone orally 6-16 hours before the run-in dose and one or more step doses in cycle 1

[0255] 8 mg / kg tocilizumab administered intravenously 1 hour before the run-in dose and one or more step doses in cycle 1

[0256] Etanercept 50 mg subcutaneously on Day -2 and then 2 days before the run-in dose and one or more step doses of Cycle 1 (except the step dose on Day 4 in the case of two-step or three-step dosing)

[0257] Acetaminophen 650 mg orally 1 hour before the run-in dose and one or more step doses in cycle 1

[0258] The above CRS mitigation strategies will initially be administered using intravenous dexamethasone. Based on the emerging safety profile, intravenous dexamethasone may be discontinued while continuing only one of the above CRS strategies to evaluate the safety profile without steroid premedication.

[0259] Each subject enrolled in Part C will receive only one of the additional CRS mitigation strategies described above in addition to their AMG 757 therapy as described above. Subjects will begin taking a dose of AMG 757 that is considered safe and tolerable. If the incidence of CRS decreases based on emerging safety data, one or more of the above strategies may be implemented into Part A or B.

[0260] Table 2 summarizes the eligibility criteria for study 20160323

[0261] Table 2 Key eligibility criteria

[0262]

[0263]

[0264] Example 4 Study Endpoints of Trial 20160323

[0265] The hypothesis of Study 20160323 was that AMG 757 would be safe and tolerable in subjects with Indications A and B.

[0266] Primary endpoints: dose-limiting toxicity (DLT), treatment-emergent adverse events (AEs), treatment-related AEs, and clinically significant changes in vital signs, ECG, physical examination, and clinical laboratory tests.

[0267] Secondary endpoints: For indications A and B: (1) PK parameters of AMG 757 after intravenous administration, including but not limited to maximum observed concentration (C max ), minimum observed concentration (C min ), area under the concentration-time curve (AUC) over a 2-week dosing interval, accumulation after multiple dosing, and, if available, half-life (t1 / 2 ), (2) objective response (OR) according to modified Response Evaluation Criteria in Solid Tumors (RECIST) 1.1, (3) duration of response (DOR), and (4) 1-year progression-free survival (PFS) and (5) 1-year overall survival (OS). For indication B only: relapse-free survival (RFS).

[0268] Exploratory endpoints: For Indications A and B: (1) incidence of anti-AMG 757 antibody formation, (2) changes in protein, nucleic acid, and cellular biomarkers (e.g., cytokines, lymphocyte status, CTCs, sDLL3) in blood; (3) cell surface protein expression (e.g., DLL3) and tumor-infiltrating lymphocyte status in tumor tissue at baseline. For Indication B only: effect of prior chemotherapy on T-cell factor production before AMG 757 treatment. For Part C only: incidence of CRS.

[0269] Example 5: The role of people

[0270] Pharmacokinetics in humans: A preliminary PK analysis of AMG 757 was performed in subjects in cohorts 1-5 using a non-compartmental model approach. Preliminary PK results showed that AMG 757 exposure increased with dose from 0.003 mg to 0.3 mg. After multiple Q2W dosing, the estimated terminal half-life (t 1 / 2 ) was approximately one week. No significant accumulation (<2-fold) was observed.

[0271] There was one confirmed response at 0.3 mg IV Q2W (Group 5).

[0272] The following is a summary of the AMG 757 doses and dosing regimens used in the different cohorts in the study.

[0273]

[0274] Antitumor activity was assessed using modified Response Evaluation Criteria in Solid Tumors (RECIST) 1.1; assessments were performed at screening and every 8±1 weeks after AMG757 treatment until disease progression, withdrawal of consent, or initiation of new anticancer therapy.

[0275] Forty patients (median age [range], 64 years [44-80]; ECOG PS: 0-1, n=39 [97.5%], median prior line: 2.0 [1-6]; prior PD-1 / PD-L1 therapy: n=17 [42.5%]) enrolled at eight dose levels (DL) received ≥1 dose of AMG 757. The median treatment duration was 6.1 weeks (0.1-59.4). Adverse events (AEs) occurred in 39 (97.5%) patients, leading to discontinuation in 4 (10.0%); 32 (80.0%) were treatment-related, including 7 (17.5%) ≥ Grade 3 and 1 (2.5%) Grade 5 (pneumonitis; DL5 [0.3 mg]). Cytokine release syndrome (CRS) was reported in 18 (45.0%) patients; grade 2 CRS in 5 (12.5%); no CRS was ≥ grade 3. CRS was primarily manifested by fever ± hypotension, was reversible, did not lead to treatment interruption or discontinuation, occurred primarily within 24 hours of the first two doses of AMG 757, and was managed with supportive care, corticosteroids, and / or anti-IL-6 therapy. Six (17.6%) patients developed treatment-emergent anti-AMG 757 binding antibodies. Anti-AMG 757 antibodies were not associated with AEs and had no clear effect on drug exposure. Figure 5 The mean (+SD) steady-state serum concentration-time curves for AMG 757 are shown in Figure 2. AMG 757 showed a dose-proportional increase in exposure ( Figure 5 ).

[0276] Confirmed partial responses (PRs) were reported in 6 (15.8%) patients (1 / 12 [8.3%] in DL5, 1 / 8 [12.5%] in DL6, 3 / 7 [42.9%] in DL7, and 1 / 7 [14.3%] in DL8). Figure 4 ]). Stable disease was reported in 11 (28.9%), including 1 patient with an ongoing unconfirmed PR in DL8. Patients with confirmed PR had a median of 2 (1-4) prior lines of therapy, and duration of response ranged from 1.9+ to 9.4+ months. DLL3 expression at any level was observed in 31 / 32 (96.9%) of patient tumor samples, with an overall H-score of 40-300. Tumor shrinkage occurred across a wide range of DLL3 expression (H-score, 55-300). The results suggest that AMG 757 has an acceptable safety profile at doses up to 10 mg and shows antitumor activity in patients with SCLC.

[0277] Fifty-nine patients enrolled across ten dose levels (DL) received ≥1 dose of AMG 757. Figure 6A summary of objective responses is shown for patients who met the criteria for the overall response rate (ORR) analysis at the relevant data cutoff date. Confirmed partial responses (PRs) were reported in 7 (14.6%) patients (1 / 12 [8.3%] in DL5, 1 / 8 [12.5%] in DL6, 3 / 7 [42.9%] in DL7, 2 / 10 [20%] in DL8, and 0 / 7 [0%] in DL9 [ Figure 6 ]). Stable disease was reported in 12 patients (25%), including two patients in cohort 9 (2 / 7 [28.6%]). In cohort 9, grade 3 CRS was observed in one patient and was managed with supportive care, corticosteroids, and vasopressor therapy. The results showed that AMG 757 had an acceptable safety profile at doses up to 30 mg and showed antitumor activity in patients with SCLC.

[0278] Seven patients enrolled in Cohort 10 received ≥1 dose of AMG 757, with 4 subjects receiving 100 mg at least once (Day 8). No new safety signals were observed in these patients compared to earlier cohorts.

[0279] According to the teaching of the references cited in the specification, this specification will be most thoroughly understood. The embodiments in the specification provide demonstrations of embodiments of the present invention, and should not constitute limitations on the scope of the invention. It will be readily appreciated by the technician that many other embodiments are encompassed by the present invention. All publications, patents and sequences cited in this disclosure are combined by reference in their entirety. When the material incorporated by reference conflicts or is inconsistent with this specification to a certain extent, this specification will replace any such material. The citation of any reference herein is not to admit that such reference is prior art of the present invention.

[0280] Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation many equivalents to the specific embodiments of the invention described herein. Such equivalent embodiments are intended to be encompassed by the following embodiments.

[0281] Table 3 Sequence Listing

[0282]

[0283]

[0284]

[0285]

[0286]

[0287]

[0288]

[0289]

[0290]

[0291]

[0292]

[0293]

[0294]

[0295]

[0296]

[0297]

[0298]

[0299]

[0300]

[0301]

[0302]

[0303]

[0304]

[0305]

[0306]

[0307]

[0308]

[0309]

[0310]

[0311]

[0312]

[0313]

[0314]

[0315]

[0316]

[0317]

[0318]

[0319]

[0320]

[0321]

[0322]

[0323]

[0324]

[0325]

[0326]

[0327]

[0328]

[0329]

[0330]

[0331]

[0332]

[0333]

[0334]

[0335]

[0336]

[0337]

[0338]

Claims

1. Use of an anti-DLL3 agent comprising the amino acid sequence of SEQ ID NO: 438 in the preparation of a medicament for treating DLL3-positive extensive-stage small cell lung cancer (SCLC) in a human subject in need thereof, wherein the anti-DLL3 agent is a bispecific antibody construct that binds human DLL3 and human CD3 and comprises a DLL3 binding domain, the DLL3 binding domain comprising (a) a heavy chain variable region (VH) comprising: (i) a VH complementarity determining region 1 (CDR-H1) consisting of the amino acid sequence of SEQ ID NO: 31; (ii) a CDR-H2 consisting of the amino acid sequence of SEQ ID NO: 32; and (iii) a CDR-H3 consisting of the amino acid sequence of SEQ ID NO: 33; and (b) a light chain variable region (VL) comprising: (i) a VL complementarity determining region 1 (CDR-L1) consisting of the amino acid sequence of SEQ ID NO: 34; (ii) a CDR-L2 consisting of the amino acid sequence of SEQ ID NO: NO:35; and (iii) CDR-L3, which consists of the amino acid sequence of SEQ ID NO:36, and the medicament is formulated to administer the anti-DLL3 agent once every two weeks at a dose of from 10 mg to 100 mg.

2. The use of claim 1, wherein the medicament is formulated for administration of the anti-DLL3 agent at a dose of 10 mg, 30 mg or 100 mg once every two weeks.

3. The use of claim 1 or claim 2, wherein the medicament is formulated to administer the anti-DLL3 agent on days 1 and 15 of a 28-day cycle.

4. The use of claim 1 or claim 2, wherein the medicament is formulated for administration of the anti-DLL3 agent by intravenous (IV) infusion.

5. The use of claim 1 or claim 2, wherein the cancer is relapsed / refractory (RR SCLC).

6. The use of claim 1 or claim 2, wherein the anti-DLL3 agent comprises the amino acid sequence of SEQ ID NO:

520.

7. Use of an anti-DLL3 agent comprising the amino acid sequence of SEQ ID NO: 438 in the preparation of a kit for treating DLL3-positive extensive-stage small cell lung cancer (SCLC) in a human subject in need thereof, wherein the anti-DLL3 agent is a bispecific antibody construct that binds human DLL3 and human CD3 and comprises a DLL3 binding domain, the DLL3 binding domain comprising (a) a heavy chain variable region (VH) comprising: (i) a VH complementarity determining region 1 (CDR-H1) consisting of the amino acid sequence of SEQ ID NO: 31; (ii) a CDR-H2 consisting of the amino acid sequence of SEQ ID NO: 32; and (iii) a CDR-H3 consisting of the amino acid sequence of SEQ ID NO: 33; and (b) a light chain variable region (VL) comprising: (i) a VL complementarity determining region 1 (CDR-L1) consisting of the amino acid sequence of SEQ ID NO: 34; (ii) a CDR-L2 consisting of the amino acid sequence of SEQ ID NO: NO:35; and (iii) CDR-L3, which consists of the amino acid sequence of SEQ ID NO:36, and the kit comprises a drug formulated to administer the anti-DLL3 agent once every two weeks in a 28-day cycle at a dose from 10 mg to 100 mg.

8. The use of claim 7, wherein the kit is used in combination with saline, an anti-inflammatory agent, tocilizumab or etanercept, which are administered to the subject in a first cycle of administration of the anti-DLL3 agent.

9. The use of claim 7, wherein the kit further comprises saline and wherein 1 L of saline is administered to the subject by IV infusion.

10. The use of claim 9, wherein 1 L of saline is administered by IV infusion over 4 to 5 hours.

11. The use of claim 7, wherein the kit further comprises an anti-inflammatory agent, the anti-inflammatory agent being a corticosteroid or acetaminophen.

12. The use according to claim 7, wherein the kit further comprises an anti-inflammatory agent, and the anti-inflammatory agent is dexamethasone.

13. The use of claim 12, wherein the anti-DLL3 agent is administered via a 1 hour intravenous (IV) infusion.

14. Use of an anti-DLL3 agent comprising the amino acid sequence of SEQ ID NO: 438 in combination with saline, an anti-inflammatory agent, tocilizumab or etanercept in the manufacture of a kit for treating DLL3-positive extensive-stage small cell lung cancer (SCLC) in a human subject in need thereof, wherein the anti-DLL3 agent is a bispecific antibody construct that binds human DLL3 and human CD3 and comprises a DLL3 binding domain comprising (a) a heavy chain variable region (VH) comprising: (i) a VH complementarity determining region 1 (CDR-H1) consisting of the amino acid sequence of SEQ ID NO: 31; (ii) a CDR-H2 consisting of the amino acid sequence of SEQ ID NO: 32; and (iii) a CDR-H3 consisting of the amino acid sequence of SEQ ID NO: 33; and (b) a light chain variable region (VL) comprising: (i) a VL complementarity determining region 1 (CDR-L1) consisting of the amino acid sequence of SEQ ID NO:

34. NO:34; (ii) CDR-L2, which consists of the amino acid sequence of SEQ ID NO:35; and (iii) CDR-L3, which consists of the amino acid sequence of SEQ ID NO:36, and the kit is formulated to administer the anti-DLL3 agent once every two weeks in a 28-day cycle at a dose of from 10 mg to 100 mg and administer the saline, anti-inflammatory agent, tocilizumab or etanercept to the subject in the first cycle of administering the anti-DLL3 agent.

15. The use according to any one of claims 7 to 14, wherein the cancer is relapsed / refractory SCLC (RRSCLC).

16. The use of any one of claims 7-14, wherein the anti-DLL3 agent comprises the amino acid sequence of SEQ ID NO:

520.

17. The use of claim 15, wherein the anti-DLL3 agent comprises the amino acid sequence of SEQ ID NO:

520.

18. Use of an anti-DLL3 agent comprising the amino acid sequence of SEQ ID NO: 438 in the preparation of a kit for treating DLL3-positive extensive-stage small cell lung cancer (SCLC) in a human subject in need thereof, wherein the anti-DLL3 agent is a bispecific antibody construct that binds human DLL3 and human CD3 and comprises a DLL3 binding domain, the DLL3 binding domain comprising (a) a heavy chain variable region (VH) comprising: (i) a VH complementarity determining region 1 (CDR-H1) consisting of the amino acid sequence of SEQ ID NO: 31; (ii) a CDR-H2 consisting of the amino acid sequence of SEQ ID NO: 32; and (iii) a CDR-H3 consisting of the amino acid sequence of SEQ ID NO: 33; and (b) a light chain variable region (VL) comprising: (i) a VL complementarity determining region 1 (CDR-L1) consisting of the amino acid sequence of SEQ ID NO: 34; (ii) a CDR-L2 consisting of the amino acid sequence of SEQ ID NO: NO:35; and (iii) CDR-L3, which consists of the amino acid sequence of SEQ ID NO:36, and the kit comprises a drug formulated to administer the anti-DLL3 agent according to the following regimen: a) First dose from 0.5 mg to 2 mg on day 1, b) a second dose from 3 mg to 100 mg on day 8, and c) one or more subsequent doses from 3 mg to 100 mg starting on day 15 and every two weeks thereafter, and wherein the second dose and the subsequent dose are the same, and wherein the kit further comprises an anti-inflammatory agent formulated to be administered on day 1 prior to the first dose of the anti-DLL3 agent and on day 8 prior to the second dose of the anti-DLL3 agent.

19. The use of claim 18, wherein the first dose is 1 mg, and the second dose and the subsequent dose are each 3 mg, 10 mg, 30 mg or 100 mg.

20. The use of claim 19, wherein the second dose and the subsequent dose are 10 mg.

21. The use of claim 18, wherein the anti-DLL3 agent is administered in a 28-day cycle and the kit is used in combination with saline, tocilizumab or etanercept, which is administered to the subject in the first cycle in which the anti-DLL3 agent is administered.

22. The use of claim 21, wherein the kit further comprises saline, and wherein one liter of saline is administered to the subject by IV infusion following the first and second doses of the anti-DLL3 agent.

23. The use of claim 22, wherein one liter of saline is administered by IV infusion over 4 to 5 hours.

24. The use of claim 22, wherein one liter of saline is administered by IV infusion on day 15.

25. The use of claim 23, wherein one liter of saline is administered by IV infusion on day 15.

26. The use of claim 21, wherein the kit comprises tocilizumab and tocilizumab is administered to the subject by IV infusion prior to the first and second doses of the anti-DLL3 agent.

27. The use of claim 18, wherein the kit comprises an anti-inflammatory agent which is a corticosteroid or acetaminophen.

28. The use of claim 18, wherein the kit comprises an anti-inflammatory agent, and the anti-inflammatory agent is dexamethasone.

29. The use of claim 20, wherein the kit comprises an anti-inflammatory agent, and the anti-inflammatory agent is dexamethasone.

30. The use of claim 22, wherein the kit comprises an anti-inflammatory agent, and the anti-inflammatory agent is dexamethasone.

31. The use of claim 18, wherein the anti-DLL3 agent is administered by IV infusion.

32. The use of claim 18, wherein the anti-DLL3 agent is administered via a one-hour intravenous (IV) infusion.

33. The use of any one of claims 18-32, wherein the cancer is relapsed / refractory SCLC (RRSCLC).

34. The use of any one of claims 18-32, wherein the anti-DLL3 agent comprises the amino acid sequence of SEQ ID NO:

520.

35. The use of claim 33, wherein the anti-DLL3 agent comprises the amino acid sequence of SEQ ID NO:

520.

36. Use of an anti-DLL3 agent comprising the amino acid sequence of SEQ ID NO: 438 in the preparation of a kit for treating DLL3-positive extensive-stage small cell lung cancer (SCLC) in a human subject in need thereof, wherein the anti-DLL3 agent is a bispecific antibody construct that binds human DLL3 and human CD3 and comprises a DLL3 binding domain, the DLL3 binding domain comprising (a) a heavy chain variable region (VH) comprising: (i) a VH complementarity determining region 1 (CDR-H1) consisting of the amino acid sequence of SEQ ID NO: 31; (ii) a CDR-H2 consisting of the amino acid sequence of SEQ ID NO: 32; and (iii) a CDR-H3 consisting of the amino acid sequence of SEQ ID NO: 33; and (b) a light chain variable region (VL) comprising: (i) a VL complementarity determining region 1 (CDR-L1) consisting of the amino acid sequence of SEQ ID NO: 34; (ii) a CDR-L2 consisting of the amino acid sequence of SEQ ID NO: NO:35; and (iii) CDR-L3, which consists of the amino acid sequence of SEQ ID NO:36, and the kit comprises a drug formulated to administer the anti-DLL3 agent according to the following regimen: a) First dose from 0.3 mg to 3 mg on day 1, b) a second dose from 10 mg to 100 mg on day 8, and c) one or more subsequent doses from 10 mg to 100 mg starting on day 15 and every two weeks thereafter, and wherein the second dose and the subsequent dose are the same.

37. The use of claim 36, wherein the first dose is from 1 mg to 3 mg and The second and subsequent doses are from 10 mg to 15 mg.

38. The use of claim 36, wherein the anti-DLL3 agent is administered by IV infusion.

39. The use of claim 36, wherein the kit further comprises an anti-inflammatory agent.

40. The use of claim 39, wherein the anti-inflammatory agent is a corticosteroid.

41. The use of claim 40, wherein the corticosteroid is dexamethasone.

42. The use of claim 36, wherein the anti-DLL3 agent is administered in a 28-day cycle and the kit is used in combination with saline, tocilizumab or etanercept, which is administered to the subject in the first cycle of administration of the anti-DLL3 agent.

43. The use of claim 42, wherein after the first and second doses of the anti-DLL3 agent, one liter of saline is administered to the subject by IV infusion.

44. The use of any one of claims 36-43, wherein the cancer is relapsed / refractory SCLC (RRSCLC).

45. The use of any one of claims 36-43, wherein the anti-DLL3 agent comprises the amino acid sequence of SEQ ID NO:

520.

46. ​​The use of claim 44, wherein the anti-DLL3 agent comprises the amino acid sequence of SEQ ID NO: 520.

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