Anti-ceacam5 antibody drug conjugates for the treatment of cancer

Abstract

The present application relates to methods of treating cancer by administering a therapeutically effective dose of antibody drug conjugates of the formula (I) comprising anti- CEACAM5 antibodies, antigen binding portions thereof that are conjugated with a linker and exatecan. The methods comprising administering the antibody drug conjugates described herein may be suitable for combination therapy with an anti-VEGF treatment.

Description

[0001] Docket No. 14887-WO

[0002] ANTI-CEACAM5 ANTIBODY DRUG CONJUGATES

[0003] FOR THE TREATMENT OF CANCER

[0004] CROSS-REFERENCE TO RELATED APPLICATIONS

[0005]

[0001] This application claims the benefit of U.S. provisional patent application serial number 63 / 720.489, filed November 14, 2024, and U.S. provisional patent application serial number 63 / 827,494, filed June 20, 2025, each of which is incorporated herein by reference in its entirety.

[0006] REFERENCE TO SEQUENCE LISTING SUBMITTED ELECTRONICALLY

[0007]

[0002] The content of the electronically submitted sequence listing (Name: 14887-WO-PCT Sequence Listing.xmk Size: 21 kilobytes; and Date of Creation: November 3. 2025), filed with the application, is incorporated herein by reference in its entirety.

[0008] FIELD OF DISCLOSURE

[0009]

[0003] The disclosure provides methods and kits comprising a treatment of antibody drug conjugates (ADCs) comprising antibodies or antigen binding portions that specifically bind to carcinoembryonic antigen-related cell adhesion molecule-5 (CEACAM5), or combination therapies, for use in a cancer therapy.

[0010] BACKGROUND

[0011]

[0004] Recurrent or refractory advanced cancer remains a major health problem worldwide, and, according to the World Health Organization (WHO), ranks first or second to cardiovascular disease as an overall cause of mortality. Although there has been significant progress in treating a variety of malignancies over the last few decades, patients with recunent or refractory advanced solid tumors still have a poor prognosis. These patients have a high unmet medical need.

[0012]

[0005] Antibody-drug conjugates (ADC) have show n significant clinical efficacy in a wide range of hematologic and solid tumors leading to regulatory approvals. ADCs are designed to deliver the drug payload directly to tumors. CEACAM5 is a cell surface glycoprotein repressed in normal adult human gastrointestinal and respiratory tissue but Docket No. 14887-WO

[0013] present on tissue w ith malignant transformation providing the rationale for targeted delivery' of a cytotoxic pay load. In addition, circulating carcinoembryonic antigen, CEA (a synony m for CEACAM5) is a validated, FDA- approved biomarker for cancer often clinically used as a predictive marker of disease progression and survival.

[0014]

[0006] ADCs are in clinical development for treatment of solid tumors with CEACAM5 expression. However, there are no FDA-approved CEACAM5-targeted therapies for cancer. ADCs have shown significant clinical efficacy in a wide range of hematologic and solid tumors with high unmet medical need. Thus, there is a need for effective treatments for CEACAM5-expressing cancers.

[0015] SUMMARY

[0016]

[0007] An aspect of the disclosure provides a method of treating a CEACAM5 -expressing cancer in a subject in need thereof, wherein the method comprises administering a therapeutically effective dose of an ADC having the formula (I):

[0017]

[0018] or a pharmaceutically acceptable salt, a stereoisomer, or a solvate thereof, wherein n ranges from 3 to 8; represents that the configuration of the double bond may be E or Z; AB is an anti-CEAC AM5 antibody or antigen binding portion thereof comprising (a) a heavy chain variable region (VH) comprising complementarity determining region (CDR)l. CDR2, and CDR3 regions comprising the amino acid sequences set forth in SEQ ID NOs: 1, 2, and 3, respectively, and a light chain variable region (VL) comprising CDR1 , CDR2, and CDR3 regions comprising the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively.

[0019] The ADC having the formula (I): Docket No. 14887-WO

[0020]

[0021] wherein n ranges from 3 to 8,

[0022] is also understood to be identical to the ADC having the formula:

[0023]

[0024]

[0008] As the two structures are presumed to be identical, the two representations are interchangeable. Either structure can be used to represent the same structure. Likew ise, when n is 8, the structure

[0025]

[0026] ADC001,

[0027] is also identical to the structure Docket No. 14887-WO

[0028]

[0029] , and the two structures are interchangeable. Another aspect of the disclosure provides a method of treating a CEACAM5-expressing cancer in a subject in need thereof, wherein the method comprises administering a therapeutically effective dose of an ADC having the structure:

[0030]

[0031] ADC001,

[0032] or a pharmaceutically acceptable salt, a stereoisomer, or a solvate thereof, wherein ' / vwrepresents that the configuration of the double bond may be E or Z; AB is an anti-CEACAM5 antibody or antigen binding portion thereof comprising (a) a heavy chain variable region (VH) comprising complementarity determining region (CDR)l, CDR2, and CDR3 regions comprising the amino acid sequences set forth in SEQ ID NOs: 1, 2, and 3, respectively, and a light chain variable region (VL) comprising CDR1, CDR2, and CDR3 regions comprising the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively. In various embodiments, the ADC is ADC001.

[0033]

[0010] In an embodiment, the method comprises administering the ADC at a dose. In an embodiment of the method, the ADC is administered at a dose ranging from about 0.5 mg / kg body weight to about 8 mg / kg body weight. In an embodiment, the ADC could be administered at an intermediate or higher dose based on the safety and tolerability of Docket No. 14887-WO

[0034] administered doses. In an embodiment of the method, the ADC is administered at a dose ranging from about 0.5 mg / kg body weight to about 6 mg / kg body weight. In an embodiment of the method, the ADC is administered at a dose ranging from about 1 mg / kg body weight to about 6 mg / kg body weight. In an embodiment of the method, the ADC is administered at a dose ranging from about 1 mg / kg body weight to about 4 mg / kg body weight.

[0035] [on] As used herein, numerical values are often presented in a range format throughout this document. The use of a range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention unless the context clearly indicates otherwise. Accordingly, the use of a range expressly includes all possible subranges, all individual numerical values within that range, and all numerical values or numerical ranges including integers within such ranges and fractions of the values or the integers within ranges unless the context clearly indicates otherwise.

[0036]

[0012] In an embodiment, the method comprises administering the ADC at a dose. In an embodiment of the method, the ADC is administered at a dose ranging from about 0.5 mg / kg body weight to about 1 mg / kg body weight. In an embodiment of the method, the ADC is administered at a dose ranging from about 1 mg / kg body weight to about 2 mg / kg body weight. In an embodiment of the method, the ADC is administered at a dose ranging from about 2 mg / kg body weight to about 3 mg / kg body weight. In an embodiment, the ADC is administered at about 2.8 mg / kg body weight. In an embodiment of the method, the ADC is administered at a dose ranging from about 3 mg / kg body weight to about 4 mg / kg body weight. In an embodiment of the method, the ADC is administered at a dose ranging from about 4 mg / kg body weight to about 5 mg / kg body weight. In an embodiment of the method, the ADC is administered at a dose ranging from about 5 mg / kg body weight to about 6 mg / kg body weight. In an embodiment of the method, the ADC is administered at a dose ranging from about 6 mg / kg body weight to about 7 mg / kg body weight. In an embodiment of the method, the ADC is administered at a dose ranging from about 7 mg / kg body weight to about 8 mg / kg body weight. For example, the dose of ADC administered by the method is about 7.8 mg / kg body weight. In an embodiment of the method, the ADC is administered at a dose of 7.8 mg / kg body weight. Docket No. 14887-WO

[0037]

[0013] In an embodiment, the method comprises administering the ADC at a dose. In an embodiment, the method comprises administering the ADC at about 0.5 mg / kg body weight. In an embodiment, the method comprises administering the ADC at about 1 mg / kg body weight. In an embodiment, the method comprises administering the ADC at about 2 mg / kg body weight. In an embodiment, the method comprises administering the ADC at about 2.5 mg / kg body weight. In an embodiment, the method comprises administering the ADC at about 2.8 mg / kg body weight. In an embodiment, the method comprises administering the ADC at about 3 mg / kg body weight. In an embodiment, the method comprises administenng about 3.5 mg / kg body weight. In an embodiment, the method comprises administering the ADC at about 4 mg / kg body weight. In an embodiment, the method comprises administering about 4.5 mg / kg body weight. In an embodiment, the method comprises administering the ADC at about 5 mg / kg body weight. In an embodiment, the method comprises administering the ADC at about 6 mg / kg body weight. In an embodiment, the method comprises administering the ADC at about 7 mg / kg body weight. In an embodiment, the method comprises administering the ADC at about 8 mg / kg body weight.

[0038]

[0014] In an embodiment, the method comprises administering the ADC at a dose. In an embodiment, the method comprises administering the ADC at 0.5 mg / kg body weight. In an embodiment, the method comprises administering the ADC at 1 mg / kg body weight. In an embodiment, the method comprises administering the ADC at 2 mg / kg body weight. In an embodiment, the method comprises administering the ADC at 2.5 mg / kg body weight. In an embodiment, the method comprises administering the ADC at 2.8 mg / kg body weight. In an embodiment, the method comprises administering the ADC at 3 mg / kg body weight. In an embodiment, the method comprises administering 3.5 mg / kg body weight. In an embodiment, the method comprises administering the ADC at 4 mg / kg body weight. In an embodiment, the method comprises administering 4.5 mg / kg body weight. In an embodiment, the method comprises administering the ADC at 5 mg / kg body weight. In an embodiment, the method comprises administering the ADC at 6 mg / kg body weight. In an embodiment, the method comprises administering the ADC at 7 mg / kg body weight. In an embodiment, the method comprises administering the ADC at 8 mg / kg body weight.

[0039]

[0015] In an embodiment, the method comprises administering the ADC intravenously (IV).

[0040]

[0016] In an embodiment, the method comprises administering the ADC every two weeks (Q2W), every three weeks (Q3W), or ADC every four weeks (Q4W). For example, the Docket No. 14887-WO

[0041] method comprises administering the ADC IV in a dosing schedule that is Q2W, Q3W, or Q4W. In an embodiment, the method comprises administering the ADC Q2W. In an embodiment, the method comprises administering the ADC Q3W. In an embodiment, the method comprises administering the ADC Q4W.

[0042]

[0017] In an embodiment of the method, the ADC treatment is administered Q3W. In various embodiments, the ADC treatment is administered Q3W at a dose of about 1 mg / kg to about 6 mg / kg body weight. For example, the ADC is administered Q3W at a dose of 2.8 mg / kg body weight. In an embodiment of the method, the ADC treatment is administered Q2W. In various embodiments, the ADC treatment is administered Q2W at a dose of about 1 mg / kg to about 6 mg / kg body weight. For example, the ADC is administered Q2W at a dose of 2 mg / kg body weight. In an embodiment of the method, the ADC treatment is administered Q4W. In various embodiments, the ADC treatment is administered Q4W at a dose of about 1 mg / kg to about 6 mg / kg body weight.

[0043]

[0018] In an embodiment, the method comprises a dosing cycle of administering the ADC, wherein the dosing cycle comprises about 14 days to about 28 days. In an embodiment, the method comprises administering the ADC Q3W. In an embodiment of the method, the dosing cycle comprises about 14 days to about 21 days. In an embodiment of the method, the dosing cycle comprises about 21 days to about 28 days. In an embodiment of the method, the dosing cycle comprises about 14 days. In an embodiment of the method, the dosing cycle comprises about 21 days. In an embodiment of the method, the dosing cycle comprises about 28 days. In an embodiment of the method, the dosing cycle is described in Example 3. In an embodiment of the method, the dosing cycle is a 21 -day cycle.

[0044]

[0019] In an embodiment of the method, the anti-CEACAM5 antibody or antigen binding portion thereof comprises a VH which comprises the amino acid sequence set forth in SEQ ID NOs: 4. In an embodiment of the method, the anti-CEACAM5 antibody comprises a VH which comprises the amino acid sequence set forth in SEQ ID NOs: 4.

[0045]

[0020] In an embodiment of the method, the anti-CEACAM5 antibody or antigen binding portion thereof comprises a VL which comprises the amino acid sequence set forth in SEQ ID NO: 11. In an embodiment of the method, the anti-CEACAM5 antibody compnses a VL which comprises the amino acid sequence set forth in SEQ ID NO: 11.

[0046]

[0021] In an embodiment of the method, the anti-CEACAM5 antibody or antigen binding portion thereof comprises a VH and a VL which comprise the amino acid sequence set Docket No. 14887-WO

[0047] forth in SEQ ID NOs: 4 and the amino acid sequence set forth in SEQ ID NO: 11, respectively. In an embodiment of the method, the anti-CEACAM5 antibody comprises a VH having the amino acid sequence set forth in SEQ ID NOs: 4 and a VL having the amino acid sequence set forth in SEQ ID NO: 11, respectively.

[0048]

[0022] In an embodiment of the method, the anti-CEACAM5 antibody or antigen binding portion thereof comprises an IgGl constant region, IgG2 constant region, IgG3 constant region, IgG4 constant region, or a variant thereof. For example, the anti-CEACAM5 antibody comprises an IgGl antibody. In an embodiment of the method, the anti- CEACAM5 antibody or antigen binding portion thereof comprises an lgG1.3f constant region.

[0049]

[0023] In an embodiment of the method, the anti-CEACAM5 antibody is a human, humanized, or chimeric antibody.

[0050]

[0024] In an embodiment of the method, the anti-CEACAM5 antibody or antigen binding portion thereof comprising a heavy chain which comprises the amino acid sequence set forth in SEQ ID NOs: 6. In an embodiment of the method, the anti-CEACAM5 antibody or antigen binding portion thereof comprises a VH and a VL which comprise the amino acid sequence set forth in SEQ ID NOs: 4 and the amino acid sequence set forth in SEQ ID NO: 11, respectively. In an embodiment of the method, the anti- CEACAM5 antibody comprises a VH and a VL which comprise the amino acid sequence set forth in SEQ ID NOs: 4 and the amino acid sequence set forth in SEQ ID NO: Ir respectively.

[0051]

[0025] In an embodiment of the method, the anti-CEACAM5 antibody or antigen binding portion thereof comprises a light chain which comprises the amino acid sequence set forth in SEQ ID NO: 13. In an embodiment of the method, the anti-CEACAM5 antibody comprises a light chain which comprises the amino acid sequence set forth in SEQ ID NO: 13.

[0052]

[0026] In an embodiment of the method, the anti-CEACAM5 antibody or antigen binding portion thereof comprising a heavy chain and a light chain which comprise the amino acid sequence set forth in SEQ ID NOs: 6 and the amino acid sequence set forth in SEQ ID NO: 13. respectively. In an embodiment of the method, the anti-CEACAM5 antibody comprises a heavy chain and a light chain which comprise the amino acid sequence set forth in SEQ ID NOs: 6 and the amino acid sequence set forth in SEQ ID NO: 13, respectively. In various embodiments, the antibody comprises four polypeptide Docket No. 14887-WO

[0053] chains comprising two heavy chains and two light chains. In various embodiments, each heavy' chain comprises the amino acid sequence of SEQ ID NO: 6 and each light chain comprises the amino acid sequence of SEQ ID NO: 13. In various embodiments, each heavy chain consists of the amino acid sequence of SEQ ID NO: 6 and each light chain consists of the amino acid sequence of SEQ ID NO: 13.

[0054]

[0027] In an embodiment of the method, the antibody, or antigen binding portion thereof, comprises a bispecific molecule comprising the anti-CEACAM5 antibody or antigen binding portion thereof and a second binding region that binds to another antigen.

[0055]

[0028] In an embodiment of the method, the antibody, or antigen binding portion thereof, comprises a multi-specific molecule comprising the anti-CEACAM5 antibody, or antigen binding portion thereof and at least two binding regions, each of which binds other antigens.

[0056]

[0029] In an embodiment, the method comprises administering the ADC (comprising the anti-CEACAM5 antibody) at a dose. In various embodiments, the method comprises administering the ADC at a dose of 0.5 mg / kg body weight. In an embodiment, the method comprises administering the ADC at a dose of 1 mg / kg body weight. In an embodiment, the method comprises administering the ADC at a dose of 2 mg / kg body weight. In an embodiment, the method comprises administering the ADC at a dose of 2.5 mg / kg body weight. In an embodiment, the method comprises administering the ADC at a dose of 2.8 mg / kg body weight. In an embodiment, the method comprises administering the ADC at a dose of 3 mg / kg body weight. In an embodiment, the method comprises administering 3.5 mg / kg body weight. In an embodiment, the method comprises administering the ADC at a dose of 4 mg / kg body weight. In an embodiment, the method comprises administering 4.5 mg / kg body weight. In an embodiment, the method comprises administering the ADC at a dose of 5 mg / kg body weight. In an embodiment, the method comprises administering the ADC at a dose of 6 mg / kg body weight. In an embodiment, the method comprises administering the ADC at a dose of 7 mg / kg body weight. In an embodiment, the method comprises administering the ADC at a dose of 8 mg / kg body weight.

[0057]

[0030] An aspect of the disclosure provides a method of treating a CEACAM5 -expressing cancer in a subject in need thereof, wherein the method comprises administering a therapeutically effective dose of an anti-VEGF treatment and an ADC treatment;

[0058] wherein, Docket No. 14887-WO

[0059] (i) the anti-VEGF treatment comprises administering an anti-VEGF treatment to the subject on about day 1 of a dosing cycle; and

[0060] (ii) the ADC treatment comprises administering an ADC having the formula (I):

[0061]

[0062] or a pharmaceutically acceptable salt, a stereoisomer, or a solvate thereof, n ranges from 4 to 8; represents that the configuration of the double bond may be E or Z wherein AB is an anti-CEAC AM5 antibody or antigen binding portion thereof comprising (a) a VH comprising CDR1, CDR2, and CDR3 regions comprising the amino acid sequences set forth in SEQ ID NOs: 1, 2, and 3, respectively, and a VL comprising CDR1, CDR2, and CDR3 regions comprising the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively. In various embodiments, the ADC is formula (I).

[0063]

[0031] Another aspect of the disclosure provides a method of treating cancer in a subject in need thereof, wherein the method comprises administering a therapeutically effective dose of an anti-VEGF treatment and an ADC treatment; wherein,

[0064] (i) the anti-VEGF treatment comprises administering an anti-VEGF treatment to the subject on about day 1 of a dosing cycle; and

[0065] (ii) the ADC treatment comprises administering an ADC having the structure:

[0066]

[0067] Docket No. 14887-WO

[0068] or a pharmaceutically acceptable salt, a stereoisomer, or a solvate thereof, ' / vvvrepresents that the configuration of the double bond may be E or Z wherein AB is an anti-CEACAM5 antibody or antigen binding portion thereof comprising (a) a VH comprising CDR1, CDR2, and CDR3 regions comprising the amino acid sequences set forth in SEQ ID NOs: 1, 2, and 3, respectively, and a VL comprising CDR1, CDR2, and CDR3 regions comprising the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively.

[0069] In various embodiments, the ADC is ADC001. In an embodiment of the method, the anti-VEGF treatment is bevacizumab. In an embodiment, the method comprises administering the bevacizumab at a dose of about 5 mg / kg to about 7.5 mg / kg body weight. In an embodiment, the method comprises administering the bevacizumab at a dose of 5 mg / kg body weight. In an embodiment, the method comprises administering the bevacizumab at a dose of 7.5 mg / kg body weight. In an embodiment, the method comprises administering 5 mg / kg bevacizumab Q2W. In an embodiment, the method comprises administering 7.5 mg / kg body weight bevacizumab Q3W.

[0070]

[0032] In an embodiment, the method comprises administering the ADC at a dose. In an embodiment of the method, the ADC is administered at a dose ranging from about 0.5 mg / kg body weight to about 8 mg / kg body weight. In an embodiment of the method, the ADC is administered at a dose ranging from about 1 mg / kg body weight to about 6 mg / kg body weight. In an embodiment of the method, the ADC is administered at a dose ranging from about 1 mg / kg body weight to about 4 mg / kg body weight.

[0071]

[0033] In an embodiment, the method comprises administering the ADC at a dose. In an embodiment of the method, the ADC is administered at a dose ranging from about 0.5 mg / kg body weight to about 1 mg / kg body weight. In an embodiment of the method, the ADC is administered at a dose ranging from about 1 mg / kg body weight to about 2 mg / kg body weight. In an embodiment of the method, the ADC is administered at a dose ranging from about 2 mg / kg body weight to about 3 mg / kg body weight. In an embodiment, the ADC is administered at about 2.8 mg / kg body weight. In an embodiment of the method, the ADC is administered at a dose ranging from about 3 mg / kg body weight to about 4 mg / kg body weight. In an embodiment of the method, the ADC is administered at a dose ranging from about 4 mg / kg body weight to about 5 mg / kg body weight. In an embodiment of the method, the ADC is administered at a dose ranging from about 5 mg / kg body weight to about 6 mg / kg body weight. In an embodiment of the method, the ADC is administered at a dose ranging from about 6 mg / kg body weight to

[0072]

[0073] Docket No. 14887-WO

[0074] about 7 mg / kg body. In an embodiment of the method, the ADC is administered at a dose ranging from about 7 mg / kg body weight to about 8 mg / kg body.

[0075]

[0034] In an embodiment, the method comprises administering the ADC at a dose. In an embodiment, the method comprises administering the ADC at a dose of about 0.5 mg / kg body weight. In an embodiment, the method comprises administering the ADC at a dose of about 1 mg / kg body weight. In an embodiment, the method comprises administering the ADC at a dose of about 2 mg / kg body weight. In an embodiment, the method comprises administering the ADC at a dose of about 2.5 mg / kg body weight. In an embodiment, the ADC is administered at a dose of about 2.8 mg / kg body weight. In an embodiment, the method comprises administering the ADC at a dose of about 3 mg / kg body weight. In an embodiment, the method comprises administering the ADC a dose of at about 4 mg / kg body weight. In an embodiment, the method comprises administering the ADC at a dose of about 5 mg / kg body weight. In an embodiment, the method comprises administering the ADC at a dose of about 6 mg / kg body weight. In an embodiment, the method comprises administering the ADC at a dose of about 7 mg / kg body weight. In an embodiment, the method comprises administering the ADC at a dose of about 8 mg / kg body weight. For example, the dose of ADC administered by the method is about 7.8 mg / kg body weight. In an embodiment of the method, the ADC is administered at a dose of 7.8 mg / kg body weight.

[0076]

[0035] In an embodiment, the method comprises administering the ADC Q2W,

[0077] Q3W, or Q4W. In an embodiment, the method comprises administering the ADC Q3W.

[0078]

[0036] In an embodiment, the method comprises administering the ADC IV in a dosing schedule that is Q2W, Q3W, or Q4W. In an embodiment, the method comprises administering the ADC IV in a dosing schedule that is Q3W.

[0079]

[0037] In an embodiment, the method comprises administering the ADC on a dosing cycle that is either Q2W. Q3W, or Q4W. In certain embodiments, the dosing cycle comprises a 21 -day cycle or a 28-day cycle. In an embodiment of the method, administering the ADC on the Q3W dosing cycle comprises administering the ADC on Day 1 of the 21- day cycle. In an embodiment of the method, administering the ADC on the Q2W dosing cycle comprises administering the ADC on Day 1 and Day 15 in a 28-day cycle. In an embodiment of the method, the administering the ADC on Q4W dosing cycle comprises administering the ADC on Day 1 of the 28-day cycle.

[0080]

[0081] Docket No. 14887-WO

[0082]

[0038] In an embodiment of the method, the ADC treatment is administered Q3W. In various embodiments, the ADC treatment is administered Q3W at a dose of about 1 mg / kg to about 6 mg / kg body weight. For example, the ADC is administered Q3W at a dose of 2.8 mg / kg body weight. In an embodiment of the method, the ADC treatment is administered Q2W. In various embodiments, the ADC treatment is administered Q2W at a dose of about 1 mg / kg to about 6 mg / kg body weight. For example, the ADC is administered Q2W at a dose of 2 mg / kg body weight. In an embodiment of the method, the ADC treatment is administered Q4W. In various embodiments, the ADC treatment is administered Q4W at a dose of about 1 mg / kg to about 6 mg / kg body weight.

[0083]

[0039] In an embodiment of the method, the anti-CEACAM5 antibody or antigen binding portion thereof comprises a VH which comprises the amino acid sequence set forth in SEQ ID NOs: 4. In an embodiment of the method, the anti-CEACAM5 antibody comprises a VH which comprises the amino acid sequence set forth in SEQ ID NOs: 4.

[0084]

[0040] In an embodiment of the method, the anti-CEACAM5 antibody or antigen binding portion thereof comprises a VL which comprises the amino acid sequence set forth in SEQ ID NO: 11. In an embodiment of the method, the anti-CEACAM5 antibody comprises a VL which comprises the amino acid sequence set forth in SEQ ID NO: 11.

[0085]

[0041] In an embodiment of the method, the anti-CEACAM5 antibody or antigen binding portion thereof comprising a VH and a VL which comprise the amino acid sequence set forth in SEQ ID NOs: 4 and the amino acid sequence set forth in SEQ ID NO: 11, respectively. In an embodiment of the method, the anti-CEACAM5 antibody comprises a VH and a VL which comprise the amino acid sequence set forth in SEQ ID NOs: 4 and the amino acid sequence set forth in SEQ ID NO: 11 , respectively.

[0086]

[0042] In an embodiment of the method, the anti-CEACAM5 antibody or antigen binding portion thereof comprises an IgGl constant region, IgG2 constant region, IgG3 constant region, IgG4 constant region, or a variant thereof. For example, the anti-CEACAM5 antibody or antigen binding portion thereof comprises an IgGl antibody. In an embodiment of the method, the anti-CEACAM5 antibody or antigen binding portion thereof comprises an IgG1.3f constant region.

[0087]

[0043] In an embodiment of the method, the anti-CEACAM5 antibody is a human, humanized, or chimeric antibody.

[0088]

[0044] In an embodiment of the method, the anti-CEACAM5 antibody or antigen binding portion thereof comprising a heavy chain which comprises the amino acid sequence set

[0089]

[0090] Docket No. 14887-WO

[0091] forth in SEQ ID NO: 6. In an embodiment of the method, the anti-CEACAM5 antibody comprises a heavy chain which comprises the amino acid sequence set forth in SEQ ID NO: 6.

[0092]

[0045] In an embodiment of the method, the anti-CEACAM5 antibody or antigen binding portion thereof comprising a light chain which comprises the amino acid sequence set forth in SEQ ID NO: 13. In an embodiment of the method, the anti-CEACAM5 antibody comprises a light chain which comprises the amino acid sequence set forth in SEQ ID NO: 13.

[0093]

[0046] In an embodiment of the method, the anti-CEACAM5 antibody or antigen binding portion thereof comprising a heavy chain and a light chain which comprise the amino acid sequence set forth in SEQ ID NO: 6 and the amino acid sequence set forth in SEQ ID NO: 13, respectively. In an embodiment of the method, the anti-CEACAM5 antibody comprises a heavy chain and a light chain which comprise the amino acid sequence set forth in SEQ ID NO: 6 and the amino acid sequence set forth in SEQ ID NO: 13, respectively.

[0094]

[0047] In various embodiments, the antibody comprises four polypeptide chains comprising two heavy chains and two light chains. In various embodiments, each heavy chain comprises the amino acid sequence of SEQ ID NO: 6 and each light chain comprises the amino acid sequence of SEQ ID NO: 13. In various embodiments, each heavy chain consists of the amino acid sequence of SEQ ID NO: 6 and each light chain consists of the amino acid sequence of SEQ ID NO: 13.

[0095]

[0048] In an embodiment, the method comprises administering the ADC at a dose. In an embodiment, the method comprises administering the ADC at a dose of 0.5 mg / kg body weight. In an embodiment, the method comprises administering the ADC at a dose of 1 mg / kg body weight. In an embodiment, the method comprises administering the ADC at a dose of 2 mg / kg body weight. In an embodiment, the method comprises administering the ADC at a dose of 2.5 mg / kg body weight. In an embodiment, the method comprises administering the ADC at a dose of 2.8 mg / kg body weight. In an embodiment, the method comprises administering the ADC at a dose of 3 mg / kg body weight. In an embodiment, the method comprises administering the ADC at a dose of 3.5 mg / kg body weight. In an embodiment, the method comprises administering the ADC at a dose of 4 mg / kg body weight. In an embodiment, the method comprises administering the ADC at a dose of 4.5 mg / kg body weight. In an embodiment, the

[0096]

[0097] Docket No. 14887-WO

[0098] method comprises administering the ADC at a dose of 5 mg / kg body weight. In an embodiment, the method comprises administering the ADC at a dose of 6 mg / kg body weight. In an embodiment, the method comprises administering the ADC at a dose of 7 mg / kg body weight. In an embodiment, the method comprises administering the ADC at a dose of 8 mg / kg body weight. For example, the dose of ADC administered by the method is about 7.8 mg / kg body weight. In an embodiment of the method, the ADC is administered at a dose of 7.8 mg / kg body weight.

[0099]

[0049] In an embodiment of the method, the bevacizumab is administered IV. In an embodiment, the method comprises administenng the bevacizumab at a dose. In an embodiment of the method, the bevacizumab is administered at a dose of 5 mg / kg body weight. In an embodiment of the method, the bevacizumab is administered at a dose of 7.5 mg / kg body weight.

[0100]

[0050] In an embodiment of the method, the bevacizumab is administered Q2W or Q3W.

[0101]

[0051] In an embodiment of the method, the 7.5 mg / kg body weight of bevacizumab is administered Q3W. In an embodiment of the method, the 5.0 mg / kg body weight of bevacizumab is administered Q2W.

[0102]

[0052] In an embodiment of the method, the ADC treatment is administered Q3W. In various embodiments, the ADC treatment is administered Q3W at a dose of about 1 ing / kg to about 6 mg / kg body weight. For example, the ADC is administered Q3W at a dose of 2.8 mg / kg body weight. In an embodiment of the method, the ADC treatment is administered Q2W. In various embodiments, the ADC treatment is administered Q2W at a dose of about I mg / kg to about 6 mg / kg body weight. For example, the ADC is administered Q2W at a dose of 2 mg / kg body weight. In an embodiment of the method, the ADC treatment is administered Q4W. In various embodiments, the ADC treatment is administered Q4W at a dose of about 1 mg / kg to about 6 mg / kg body weight.

[0103]

[0053] In an embodiment, the method comprises administering the ADC on a dosing cycle that is either Q2W, Q3W or Q4W. In certain embodiments, the dosing cycle comprises a 21 -day cycle or about a 28-day cycle. In an embodiment of the method, administering the ADC on the Q3W dosing cycle comprises administering the ADC on Day 1 of 21- day cycle in Q3W schedule. In an embodiment of the method, administering the ADC on the Q2W dosing cycle comprises administering the ADC on Day 1 and Day 15 in the 28-day cycle. In an embodiment of the method, the administering the ADC on Q4W dosing cycle comprises administering the ADC on Day 1 of the 28-day cycle.

[0104]

[0105] Docket No. 14887-WO

[0106]

[0054] In an embodiment of the method, the bevacizumab is administered IV. In an embodiment of the method, the bevacizumab is administered at a dose of 5 mg / kg body weight. In an embodiment of the method, the bevacizumab is administered at a dose of 7.5 mg / kg body weight.

[0107]

[0055] In an embodiment of the method, the bevacizumab is administered Q2W or Q3W.

[0108]

[0056] In an embodiment of the method, the 7.5 mg / kg body weight of bevacizumab is administered Q3W. In an embodiment of the method, the 5.0 mg / kg body weight of bevacizumab is administered Q2W.

[0109]

[0057] In an embodiment of the method, ADC and / or bevacizumab are administered for as long as clinical benefit is observed or until unmanageable toxicity or disease progression occurs.

[0110]

[0058] In an embodiment of the method, the ADC and bevacizumab are administered for at least 104 weeks total.

[0111]

[0059] In an embodiment of the method, the ADC and / or bevacizumab are formulated for intravenous administration. In an embodiment of the method, the ADC is formulated for IV administration. In an embodiment of the method, the bevacizumab is formulated for IV administration.

[0112]

[0060] In an embodiment of the method, the ADC and bevacizumab are administered sequentially to the subject.

[0113]

[0061] In an embodiment of the method, the ADC and bevacizumab are administered sequentially to the subject during the induction phase.

[0114]

[0062] In an embodiment of the method, the ADC is administered before the bevacizumab is administered.

[0115]

[0063] In an embodiment of the method, the bevacizumab is administered before the ADC is administered.

[0116]

[0064] In an embodiment of the method, the ADC and the bevacizumab are administered concurrently in separate compositions.

[0117]

[0065] In an embodiment of the method, the ADC and the bevacizumab are admixed as a single composition for concurrent administration.

[0118]

[0066] In an embodiment of the method, the ADC and bevacizumab are administered within about 30 minutes to about 60 minutes of each other.

[0119]

[0067] In an embodiment of the method, the cancer is a solid tumor cancer. In an embodiment of the method, the cancer is selected from the group consisting of non-

[0120]

[0121] Docket No. 14887-WO

[0122] small cell lung cancer (NSCLC), gastric cancer (GC), and colorectal carcinoma (CRC). For example, the cancer is CRC.

[0123]

[0068] In an embodiment of the method, the CRC is metastatic, recurrent, or unresectable CRC. In an embodiment of the method, the CRC comprises adenocarcinoma histology treated with at least one line of systemic cancer therapy in the metastatic or adjuvant setting.

[0124]

[0069] In an embodiment of the method, the cancer is NSCLC. In an embodiment of the method, the NSCLC is histologically confirmed NSCLC meeting stage criteria for Stage 1IIB, Stage IV, or recurrent disease.

[0125]

[0070] In an embodiment of the method, the NSCLC has progressed on at least 2 prior lines of therapy.

[0126]

[0071] An aspect of the disclosure provides a kit for treating a subject afflicted with a CEACAM5-expressing cancer, the kit comprising:

[0127] (a) a dosage range of about 0.5 mg / kg body weight to about 8 mg / kg body weight an antibody drug conjugate (ADC) having the formula (I):

[0128]

[0129] or a pharmaceutically acceptable salt, a stereoisomer, or a solvate thereof, wherein n is 4 to 8;

[0130] represents that the configuration of the double bond may be E or Z; AB is an anti- CEACAM5 antibody or antigen binding portion thereof comprising (a) a VH comprising CDR1, CDR2, and CDR3 regions comprising the amino acid sequences set forth in SEQ ID NOs: 1, 2, and 3, respectively, and a VL comprising CDR1, CDR2, and CDR3 regions comprising the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10, respectively ; (b) optionally, a anti-VEF treatment, for example the anti-VEGF treatment comprising a dosage ranging from about 5 mg / kg body weight to about 7.5 mg / kg body weight of bevacizumab; and (c) instructions for treating cancer as described herein.

[0131]

[0072] Another aspect of the disclosure provides a kit for treating a subject afflicted with a

[0132]

[0133] Docket No. 14887-WO

[0134] CEACAM5-expressing cancer, the kit comprising:

[0135] (a) a dosage range of about 0.5 mg / kg body weight to about 8 mg / kg body weight an antibody drug conjugate (ADC) having the structure:

[0136]

[0137] ADC001,

[0138] or a pharmaceutically acceptable salt, a stereoisomer, or a solvate thereof, wherein represents that the configuration of the double bond may be E or Z; AB is an anti-CEACAM5 antibody or antigen binding portion thereof comprising (a) a VH comprising CDR1, CDR2, and CDR3 regions comprising the amino acid sequences set forth in SEQ ID NOs: 1, 2, and 3, respectively, and a VL comprising CDR1, CDR2, and CDR3 regions comprising the amino acid sequences set forth in SEQ ID NOs: 8, 9. and 10. respectively: (b) optionally, a anti- VEF treatment, for example the anti-VEGF treatment comprising a dosage ranging from about 5 mg / kg body weight to about 7.5 mg / kg body weight of bevacizumab; and (c) instructions for treating cancer as described herein. In various embodiments, the ADC is ADC001.

[0139]

[0073] In an embodiment of the kit, the anti-CEACAM5 antibody or antigen binding portion thereof comprises a VH which comprises the amino acid sequence set forth in SEQ ID NOs: 4. In an embodiment of the kit, the anti-CEACAM5 antibody comprises a VH which comprises the amino acid sequence set forth in SEQ ID NOs: 4.

[0140]

[0074] In an embodiment of the kit, the anti-CEACAM5 antibody or antigen binding portion thereof comprises a VL which comprises the amino acid sequence set forth in SEQ ID NO: 11. In an embodiment of the kit, the anti-CEACAM5 antibody comprises a VL which comprises the amino acid sequence set forth in SEQ ID NO: 11.

[0141]

[0075] In an embodiment of the kit, the anti-CEACAM5 antibody or antigen binding portion thereof comprises a VH and a VL which comprise the amino acid sequence set forth in SEQ ID NOs: 4 and the amino acid sequence set forth in SEQ ID NO: 11, respectively.

[0142]

[0143] Docket No. 14887-WO

[0144] In an embodiment of the kit, the anti-CEAC AM5 antibody comprises a VH and a VL which comprise the amino acid sequence set forth in SEQ ID NOs: 4 and the amino acid sequence set forth in SEQ ID NO: 11, respectively.

[0145]

[0076] In an embodiment of the kit, the anti-CEACAM5 antibody or antigen binding portion thereof comprising a heavy chain which comprises the amino acid sequence set forth in SEQ ID NOs: 6. In an embodiment of the kit, the anti-CEACAM5 antibody or antigen comprises a heavy chain which comprises the amino acid sequence set forth in SEQ ID NOs: 6.

[0146]

[0077] In an embodiment of the kit, the anti-CEACAM5 antibody or antigen binding portion thereof comprising a light chain which comprises the amino acid sequence set forth in SEQ ID NO: 13. In an embodiment of the kit, the anti-CEACAM5 antibody comprises a light chain which comprises the amino acid sequence set forth in SEQ ID NO: 13.

[0147]

[0078] In an embodiment of the kit, the anti-CEACAM5 antibody or antigen binding portion thereof comprising a heavy chain and a light chain which comprise the amino acid sequence set forth in SEQ ID NOs: 6 and the amino acid sequence set forth in SEQ ID NO: 13, respectively. In an embodiment of the kit, the anti-CEACAM5 antibody or antigen comprises a heavy chain and a light chain which comprise the amino acid sequence set forth in SEQ ID NOs: 6 and the amino acid sequence set forth in SEQ ID NO: 13, respectively. In various embodiments, the antibody comprises four polypeptide chains comprising two heavy chains and two light chains. In various embodiments, each heavy chain comprises the amino acid sequence of SEQ ID NO: 6 and each light chain comprises the amino acid sequence of SEQ ID NO: 13.

[0148]

[0079] In various embodiments of the kit, the ADC is formulated for IV administration.

[0149] BRIEF DESCRIPTION OF THE DRAWINGS

[0150]

[0080] FIGs. 1A-1D are a set of graphs showing the CEACAM5 expression in CDX tumors relative to human tumors quantified by IHC H-score (FIG. 1A) and the in vivo efficacy of ADC001 across 3 CRC CDX models NCI-H508 (H-score 242), CL40 (H-score 139) and LSI 80 (H-score 79) (FIG. IB, FIG. 1C, and FIG. ID).

[0151]

[0081] FIGs. 2A-2C are a set of graphs show ing in vivo efficacy of ADC001 in CDX models (NCI-H508 FIG.2A; Lsl80 FIG. 2B) and a PDX model (CTG-0835 (FIG. 2C) after a

[0152]

[0153] Docket No. 14887-WO

[0154] single intravenous injection with either 3 mg / kg of the ADC or twice weekly IV injection of bevacizumab at 20 mg / kg for 3 weeks, or combination of ADC 001 and bevacizumab.

[0155]

[0082] FIG. 3 is a study design schema for the Phase l / 2a first-in-human study of ADC001 as a monotherapy and a combination therapy with bevacizumab.

[0156] DETAILED DESCRIPTION

[0157]

[0083] The present disclosure provides methods of administenng an ADC of the formula (I) treating cancer.

[0158]

[0084] Definitions

[0159]

[0085] In order for the following detailed description to be readily understood, certain terms are first defined. Additional definitions are provided throughout.

[0160]

[0086] As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. The use of "or" or "and" means "and / or" unless stated otherwise. Furthermore, use of the term "including" as well as other forms, such as "include," "includes," and "included," is not limiting.

[0161]

[0087] The term "about" as used herein when referring to a measurable value such as an amount, a temporal duration and the like, encompasses variations of up to ± 10% from the specified value. Unless otherwise indicated, all numbers expressing e.g., quantities of ingredients or properties (e.g., molecular weight, reaction conditions) described herein are to be understood as being modified by the term "about".

[0162]

[0088] As used herein, “and / or” is to be taken as specific disclosure of each of the two specified features or components with or without the other. Thus, the term “and / or” as used in a phrase such as “A and / or B” includes “A and B,” “A or B,” “A” alone, and “B” alone. Likewise, the term “and / or” as used in a phrase such as “A, B, and / or C” encompasses each of the following: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A alone; B alone; and C alone.

[0163]

[0089] As used herein, recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. For example, if a concentration range is stated as 1% to 50%, it is intended that values such as 2% to

[0164]

[0165] Docket No. 14887-WO

[0166] 40%, 10% to 30%, or 1% to 3% are expressly enumerated in this specification. These are only examples of what is specifically intended, and all possible combinations of numerical values between and including the lowest value and the highest value enumerated are to be considered to be expressly stated in this disclosure.

[0167]

[0090] The term '‘between” as used in a phrase as such '‘between A and B” or ‘'between A-B” refers to a range including both A and B.

[0168]

[0091] The term “antibody” as used to herein includes whole antibodies and any antigen binding portions (i.e., “antigen-binding portions”) or single chains thereof. An “antibody” refers, in one aspect, to a glycoprotein comprising at least two heavy (H) chains and two light (L) chains inter-connected by disulfide bonds, or an antigen binding portion thereof. Each heavy chain is comprised of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. In certain naturally occurring antibodies, the heavy chain constant region is comprised of three domains, CHI, CH2, and CH3. In certain naturally occurring antibodies, each light chain is comprised of a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region is comprised of one domain, CL. The VH and VL regions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDR), interspersed with regions that are more conserved, termed framework regions (FR). Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminus to carboxy -terminus in the following order: FR1, CDR1, FR2, CDR2, FR3. CDR3, FR4. The variable regions of the heavy and light chains contain a binding domain that interacts with an antigen. The constant regions of the antibodies may mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g, effector cells) and the first component (Clq) of the classical complement system.

[0169]

[0092] Antibodies typically bind specifically to their cognate antigen with high affinity, reflected by a dissociation constant (Ko) of 10'5to 10'11M or less. Any KD greater than about 10'4M is generally considered to indicate nonspecific binding. As used herein, an antibody that "binds specifically" to an antigen refers to an antibody that binds to the antigen and substantially identical antigens with high affinity, which means having a KD of IO'7M or less, 10'8M or less, 1 x 10'9M or less, 1 x 1010M or less, or 1 x 10'11M or less. In some aspects, the antibody specifically binds to an antigen with a KD between

[0170]

[0171] Docket No. 14887-WO

[0172] ICT8M and IO'10M or between 10'9M and IO'11M, but does not bind with high affinity to unrelated antigens.

[0173]

[0093] An “antibody” according to the present disclosure includes, but is not limited to, naturally and non-naturally occurring antibodies, monoclonal antibodies, polyclonal antibodies, chimeric antibodies, humanized antibodies, human antibodies, nonhuman antibodies, bivalent antibodies, bispecific antibodies, multispecific antibodies, single chain antibodies, diabodies, and nanobodies.

[0174]

[0094] An “isolated antibody,” as used herein, refers to an antibody which is substantially free of other antibodies having different antigenic specificities.

[0175]

[0095] The phrase “antigen binding portion” of an antibody, as used herein, refers to one or more fragments of an antibody that retain the ability to specifically bind to an antigen (e.g., human and / or cynomolgus CEACAM5). It has been shown that the antigenbinding function of an antibody can be performed by fragments of a full-length antibody. Examples of binding fragments of an antibody include (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CHI domains; (ii) a F(ab')2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) an Fd fragment consisting of the VH and CHI domains; (iv) an Fv fragment consisting of the VL and VH domains of a single arm of an antibody, (v) a dAb fragment (Ward et al., (1989) Nature 341:544-546), which consists of a VH domain; and (vi) an isolated complementarity determining region (CDR) or (vii) a combination of two or more isolated CDRs which may optionally be joined by a synthetic linker. Furthermore, although the two domains of the Fv fragment, VL and VH, are coded for by separate genes, they can be joined, using recombinant methods, by a synthetic linker that enables them to be made as a single protein chain in which the VL and VH regions pair to form monovalent molecules (known as single chain Fv (scFv); see e.g, Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883). Such single chain antibodies are also intended to be encompassed within the term “antigen-binding portion” of an antibody. Antigenbinding portions can be produced by recombinant DNA techniques, or by enzymatic or chemical cleavage of intact immunoglobulins.

[0176]

[0096] Antibody fragments within the scope of the present invention also include F(ab’)2 fragments which may be produced by enzy matic cleavage of an IgG by, for example, pepsin. Fab fragments may be produced by. for example, reduction of F(ab’)2

[0177]

[0178] Docket No. 14887-WO

[0179] with dithiothreitol or mercaptoethylamine. A Fab fragment is a VL-CL chain appended to a VH-CH1 chain by a disulfide bridge. A F(ab’)2 fragment is two Fab fragments which, in turn, are appended by two disulfide bridges. The Fab portion of an F(ab')2 molecule includes a portion of the Fc region between which disulfide bridges are located.

[0180]

[0097] As used herein, “isotype” refers to the antibody class (e.g., IgG (including IgGl, IgG2, IgG3, and IgG4), IgM, IgA (including IgAl and IgA2), IgD, and IgE antibody) that is encoded by the heavy chain constant region genes of the antibody.

[0181]

[0098] An antibody may be from any of the commonly known isotypes, including but not limited to IgA, secretory IgA, IgG and IgM. The IgG isotype is divided in subclasses in certain species: IgGl, IgG2, IgG3 and IgG4 in humans, and IgGl, IgG2a, IgG2b and IgG3 in mice. Immunoglobulins, e.g, IgGl, exist in several allotypes, which differ from each other in at most a few amino acids.

[0182]

[0099] As used herein, the term “allotype” refers to naturally occurring variants within a specific isotype group, where the variants differ in a few amino acids. Anti-CEACAM5 antibodies described herein can be of any allotype. Antibodies referred to herein as “IgG1.3f’ are IgGl antibodies of the allotype “f,” i.e., having 214R. 356E and 358M according to the EU index. A triple mutant (L234A, L235E, G237A) IgG1.3f variant comprises an amino acid sequence set forth in SEQ ID NO: 17.

[0183] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSW TVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAEGAPSVFLFPPKPKDTLMISR TPEVTCVWDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVS NKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO : 17 )

[0184]

[0100] The mutation of these residues would eliminate or decrease the binding of the antibodies to Fey receptors and / or Clq and thus reduce activator efficacy of the Fc domain of the IgGl component of an antibody.

[0185]

[0101] As used herein, the term "hypervariable region" (sometimes referred to as the “variable region”) refers to the amino acid residues of an antibody that are responsible for antigen-binding. The hypervariable region comprises amino acid residues from a "complementarity determining region" or "CDR" (e.g. residues 24-34 (CDRL1), 50-56 (CDRL2) and 89-97 (CDRL3) in the light chain variable domain and residues 31-35 (CDRH1), 50-65 (CDRH2) and 95-102 (CDRH3) in the heavy chain variable domain;

[0186]

[0187] Docket No. 14887-WO

[0188] Kabat et al., (1991) Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Sendee, National Institutes of Health, Bethesda, Md.) and / or those residues from a "hypervariable loop" (i.e. residues 26-32 (LI), 50-52 (L2) and 91-96 (L3) in the light chain variable domain and 26-32 (Hl), 53-55 (H2) and 96-101 (H3) in the heavy chain variable domain; Chothia and Lesk, (1987) J. Mol. Biol. 196: 901-917).

[0189]

[0102] As used herein, the term "framework" or "FR" residues refers to those variable domain residues other than the hypervariable region residues defined herein as CDR residues. The residue numbering above relates to the Kabat numbering system and does not necessarily correspond in detail to the sequence numbering in the accompanying Sequence Listing. Amino acid residues in antibodies can also be defined using other numbering systems, such as Chothia, enhanced Chothia, IMGT, Kabat / Chothia composite, Honegger (AHo), Contact, or any other conventional antibody numbering scheme.

[0190]

[0103] The term '‘acceptor human framework’’ refers to a framework comprising the amino acid sequence of a light chain variable domain (VL) framework, or a heavy chain variable domain (VH) framework derived from a human immunoglobulin framework or a human consensus framework. An acceptor human framework ‘'derived from” a human immunoglobulin framework or a human consensus framework may have the same amino acid sequence as the naturally occurring human immunoglobulin framework or human consensus framework, or it may have amino acid sequence changes compared to wild-type naturally occurring human immunoglobulin framework or human consensus framework. In some aspects, the number of amino acid changes are 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1. In some aspects, the VL acceptor human framework is identical in sequence to the VL human immunoglobulin framework sequence or human consensus framework sequence.

[0191]

[0104] An "Fc region,” “Fc domain.” or "Fc" refers to the C-terminal region of the heavy chain of an antibody. Thus, an Fc region comprises the constant region of an antibody excluding the first constant region immunoglobulin domain (e.g., CHI or CL).

[0192]

[0105] An "effector function" refers to the interaction of an antibody Fc region with an Fc receptor or ligand, or a biochemical event that results therefrom. Exemplaiy "effector functions" include Clq binding, complement dependent cytotoxicity (CDC), Fc receptor binding, FcyR-mediated effector functions such as ADCC and antibody dependent cell- mediated phagocytosis (ADCP), and downregulation of a cell surface receptor (e.g., the

[0193]

[0194] Docket No. 14887-WO

[0195] B cell receptor; BCR). Such effector functions generally require the Fc region to be combined with a binding domain (e.g., an antibody variable domain).

[0196]

[0106] The term “epitope’' or “antigenic determinant"’ refers to a site on an antigen (e.g. , human CEACAM5) to which an immunoglobulin or antibody specifically binds.

[0197] Epitopes can be formed both from contiguous amino acids (usually a linear epitope) or noncontiguous amino acids juxtaposed by tertiary folding of the protein (usually a conformational epitope). Epitopes formed from contiguous amino acids are typically , but not always, retained on exposure to denaturing solvents, whereas epitopes formed by tertiary folding are typically lost on treatment with denaturing solvents. An epitope ty pically includes at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or 22 amino acids in a unique spatial conformation.

[0198]

[0107] The term “monoclonal antibody,” as used herein, refers to an antibody that displays a single binding specificity and affinity for a particular epitope or a composition of antibodies in which all antibodies display a single binding specificity and affinity’ for a particular epitope. Accordingly, the term “human monoclonal antibody” refers to an antibody or antibody composition that display (s) a single binding specificity and which has variable and optional constant regions derived from human germline immunoglobulin sequences. In one aspect, human monoclonal antibodies are produced by a hybridoma which includes a B cell obtained from a transgenic non-human animal, e.g., a transgenic mouse, having a genome comprising a human heavy chain transgene and a light chain transgene fused to an immortalized cell. Monoclonal antibodies include chimeric antibodies, human antibodies, and humanized antibodies and may occur naturally or be produced recombinantly.

[0199]

[0108] The monoclonal antibodies herein also include camelized single domain antibodies.

[0200] See. e.g, Muyldermans et al. (2001) Trends Biochem. Sci. 26:230; Reichmann et al. (1999) J. Immunol. Methods 231:25; WO 94 / 04678; WO 94 / 25591; U.S. Pat. No. 6,005,079, which are hereby incorporated by reference in their entireties). In one aspect, provided herein are single domain antibodies comprising two VH domains with modifications such that single domain antibodies are formed.

[0201]

[0109] The term “recombinant antibody,” refers to antibodies that are prepared, expressed, created or isolated by recombinant means, such as (a) antibodies isolated from an animal (e.g., a mouse) that is transgenic or transchromosomal for immunoglobulin genes (e.g., human immunoglobulin genes) or a hybridoma prepared therefrom, (b)

[0202]

[0203] Docket No. 14887-WO

[0204] antibodies isolated from a host cell transformed to express the antibody, e.g., from a transfectoma, (c) antibodies isolated from a recombinant, combinatorial antibody library (e.g., containing human antibody sequences) using phage display, and (d) antibodies prepared, expressed, created or isolated by any other means that involve splicing of immunoglobulin gene sequences (e.g, human immunoglobulin genes) to other DNA sequences. Such recombinant antibodies may have variable and constant regions derived from human germline immunoglobulin sequences. In certain aspects, however, such recombinant human antibodies can be subjected to in vitro mutagenesis and thus the amino acid sequences of the VH and VL regions of the recombinant antibodies are sequences that, while derived from and related to human germline VH and VL sequences, may not naturally exist within the human antibody germline repertoire in vivo.

[0205]

[0110] A "human" antibody refers to an antibody having variable regions in which both the framework and CDR regions are derived from human germline immunoglobulin sequences. Furthermore, if the antibody contains a constant region, the constant region also is derived from human germline immunoglobulin sequences. Also encompassed are antibodies derived from human germline immunoglobulin sequences that include normal somatic hypermutations which alter the germline immunoglobulin sequences relative to the wild-type germline immunoglobulin sequences.

[0206] [Hl] A "humanized" antibody refers to an antibody in which some, most or all of the amino acids outside the CDR domains of a non-human antibody are replaced with corresponding amino acids derived from human immunoglobulins. In one aspect of a humanized form of an antibody, some, most or all of the amino acids outside the CDR domains have been replaced with amino acids from human immunoglobulins, whereas some, most or all amino acids within one or more CDR regions are unchanged. Any additions, deletions, insertions, substitutions or modifications of amino acids are permissible as long as they do not abrogate the ability of the antibody to bind to a particular antigen. A "humanized" antibody may retain an antigenic specificity' similar to that of the original antibody.

[0207]

[0112] The term “fully human antibody” refers to an antibody that comprises human immunoglobulin protein sequences only. A fully human antibody may contain murine carbohydrate chains if produced in a mouse, in a mouse cell, or in a hybridoma derived

[0208]

[0209] Docket No. 14887-WO

[0210] from a mouse cell. Similarly, “mouse antibody” refers to an antibody which comprises mouse immunoglobulin sequences only.

[0211]

[0113] A "chimeric antibody" refers to an antibody in which the variable regions are derived from one or more species and the constant regions are derived from another species, such as an antibody in which the variable regions are derived from a mouse antibody and the constant regions are derived from a human antibody. See U.S. Pat. No.

[0212] 4,816,567; and Morrison et al., (1984) Proc. Natl. Acad. Sci. USA 81: 6851-6855.

[0213]

[0114] A "domain antibody" or “nanobody” is an immunologically functional immunoglobulin fragment containing only the variable region of a heavy chain or the variable region of a light chain. In some instances, two or more Vu regions are covalently joined with a peptide linker to create a bivalent domain antibody. The two Vu regions of a bivalent domain antibody may target the same or different antigens.

[0214]

[0115] A "bivalent antibody" comprises two antigen binding sites. In some instances, the two binding sites have the same antigen specificities. However, bivalent antibodies may be bispecific.

[0215]

[0116] A “bispecific” or “bifunctional antibody” is an artificial hybrid antibody having two different heavy / light chain pairs and two different binding sites. Bispecific antibodies can be produced by a variety of methods including fusion of hybridomas or linking of Fab' fragments. See, e.g., Songsivilai & Lachmann, Clin. Exp. Immunol. 79:315-321 (1990); Kostelny et al., J. Immunol. 148, 1547-1553 (1992). Bifunctional antibodies include, for example, heterodimeric antibody conjugates (e.g.. two antibodies or antibody fragments joined together with each having different specificities), antibody / cell surface-binding molecule conjugates (e.g., an antibody conjugated to a non- antibody molecule such as a receptor), and hybrid antibodies (e.g., an antibody having binding sites for two different antigens).

[0216]

[0117] A “multispecific antibody” is an antibody (e.g, bispecific antibodies, tri-specific antibodies) that recognizes two or more different antigens or epitopes.

[0217]

[0118] As used herein, the term "single-chain Fv" or "scFv" antibody refers to antibody fragments comprising the VH and VL domains of antibody, wherein these domains are present in a single polypeptide chain. Generally, the Fv polypeptide further comprises a polypeptide linker. For a review of scFvs, see Pluckthun (1994) THE PHARMACOLOGY OF MONOCLONAL ANTIBODIES, vol. 113, Rosenburg and Moore eds. Springer-Verlag, New York, pp. 269-315.

[0218]

[0219] Docket No. 14887-WO

[0220]

[0119] The term “binds to the same epitope" is used with reference to two or more antibodies that bind to the same segment or same segments of amino acid residues. Techniques for determining whether antibodies bind to the same epitope may be determined by epitope mapping methods described herein. Other methods involve monitoring the binding of the antibody to antigen fragments (e.g.. proteolytic fragments) or to mutated variations of the antigen where loss of binding due to a modification of an amino acid residue within the antigen sequence is often considered an indication of an epitope component, such as alanine scanning mutagenesis (Cunningham & Wells (1985) Science 244:1081), yeast display of mutant target sequence variants, or analysis of chimeras. In addition, computational combinatorial methods for epitope mapping can also be employed. These methods rely on the ability' of the antibody of interest to affinity' isolate specific short peptides from combinatorial phage display peptide libraries. Antibodies having the same VH and VL or the same CDR1, 2 and 3 sequences are expected to bind to the same epitope.

[0221]

[0120] Antibodies that “compete with another antibody for binding to a target” refer to antibodies that inhibit (partially or completely ) the binding of another antibody to the target. Whether two antibodies compete with each other for binding to a target, i.e., whether and to what extent one antibody inhibits the binding of the other antibody to a target, may7be determined using known binding competition experiments involving surface plasmon resonance (SPR) and bio-layer interferometry' (BLI). In certain aspects, an antibody competes with and inhibits binding of another antibody to a target by at least 50%, 60%, 70%, 80%, 90% or 100%. The level of inhibition or competition may be different depending on which antibody is the “blocking antibody” (i.e., the antibody that when combined with an antigen blocks another immunologic reaction with the antigen). Competition assays can be conducted as described, for example, in Ed Harlow and David Lane, Cold Spring Harb. Protoc. 2006; doi: 10.1101 / pdb.prot4277 or in Chapter 11 of “Using Antibodies” by Ed Harlow7and David Lane, Cold Spring Harbor Laboratory7Press, Cold Spring Harbor, NY, USA 1999. Competing antibodies bind to the same epitope, an overlapping epitope, or to adjacent epitopes (e.g., as evidenced by steric hindrance). Two antibodies “cross-compete” if antibodies block each other both ways by at least 50%, i.e., regardless of whether one or the other antibody is contacted first with the antigen in the competition experiment.

[0222]

[0223] Docket No. 14887-WO

[0224]

[0121] Competitive binding assays for determining whether two antibodies compete or crosscompete for binding include competition for binding to cells expressing CEACAM5, e.g., by flow cytometry . Other methods include surface plasmon resonance (SPR) (e.g.. BIACORE®), solid phase direct or indirect radioimmunoassay (RIA), solid phase direct or indirect enzy me immunoassay (EIA), sandwich competition assay (see Stahli et al.. Methods in Enzymology 9:242 (1983)); solid phase direct biotin-avidin EIA (see Kirkland et al., J. Immunol. 137:3614 (1986)); solid phase direct labeled assay, solid phase direct labeled sandwich assay (see Harlow and Lane, Antibodies: A Laboratory’ Manual, Cold Spnng Harbor Press (1988)); solid phase direct label RIA using 1-125 label (see Morel et al., Mol. Immunol. 25(1):7 (1988)); solid phase direct biotin-avidin EIA (Cheung et al., Virology 176:546 (1990)); and direct labeled RIA. (Moldenhauer et al., Scand. J. Immunol. 32:77 (1990)).

[0225]

[0122] As used herein, the terms "specific binding,’" "selective binding,” "selectively binds,” and “specifically binds,” refer to antibody binding to an epitope on a predetermined antigen. Typically, the antibody (i) binds with an equilibrium dissociation constant (Kjy) of approximately less than 10'7M, such as approximately less than 10 "8M, 10’9M or IO'10M or even lower when determined by, e.g.. surface plasmon resonance (SPR) using a predetermined antigen as the analyte and the antibody as the ligand, or Scatchard analysis of binding of the antibody to antigen positive cells, and (ii) binds to the predetermined antigen with an affinity that is at least two-fold greater than its affinity for binding to a non-specific antigen (e.g., BSA, casein) other than the predetermined antigen or a closely-related antigen. Any KD greater than about 10'4M is generally considered to indicate nonspecific binding.

[0226]

[0123] The term "kassoc" or “ka,” as used herein, refers to the association rate of a particular antibody-antigen interaction, whereas the term "kdis" or “kd,” as used herein, refers to the dissociation rate of a particular antibody-antigen interaction. The term “KD,” as used herein, is intended to refer to the dissociation constant, which is obtained from the ratio of kd to ka(i.e. kd / ka) and is expressed as a molar concentration (M). KD values for antibodies can be determined using methods well established in the art. A preferred method for determining the KD of an antibody is by using surface plasmon resonance, preferably using a biosensor system such as a BIACORE® ® system or flow cytometry' and Scatchard analysis, or bio-layer interferometry.

[0227]

[0228] Docket No. 14887-WO

[0229]

[0124] The term “EC50” or “IC50” in the context of an in vitro or in vivo assay using an antibody or immunoconjugate refers to the concentration of an antibody that induces a response that is 50% of the maximal response, i.e., halfway between the maximal response and the baseline. In pharmacology, the potency of a compound is expressed as the half-maximal effective concentration (EC50), which refers to the concentration of a drug that induces a response halfway between the baseline and maximum. While expressing the potency of a compound by its EC50 value makes sense in a clinical context, it is counterintuitive in the context of bioactivity -guided purification, as the potency of a compound is inversely related to its EC50 value, and the most potent compound is the one with the lowest EC50. Half-maximal inhibitory concentration (IC50) is the most widely used and informative measure of a drug's efficacy. It indicates how much drug is needed to inhibit a biological process by half, thus providing a measure of potency of an antagonist drug in pharmacological research.

[0230]

[0125] As used herein, the term ‘‘linked” refers to the association of two or more molecules.

[0231] The linkage can be covalent or non-covalent. The linkage also can be genetic (i.e., recombinantly fused). Such linkages can be achieved using a wide variety of art recognized techniques, such as chemical conjugation and recombinant protein production.

[0232]

[0126] As used herein, the term “conjugate” is used with reference to an immunoconjugate or antibody drug conjugate comprising an anti-CEACAM5 antibody or antigen binding portion thereof described herein linked to a cytotoxic or therapeutic drug described herein.

[0233]

[0127] The term "linker," as used herein, refers to a chemical moiety comprising a covalent bond and / or any chain of atoms that may be used to covalently attach e.g., a drug to the antibody. Linkers are known in the art and include e.g., disulfide groups, thioether groups, acid labile groups, photolabile groups, peptidase labile groups and esterase labile groups. Conjugation of an antibody of the present disclosure with cytotoxic drugs or other grow th inhibitory agents may be performed e.g. using a variety’ of bifunctional protein coupling agents including but not limited to N-succinimidyl pyridyldithiobutyrate (SPDB), butanoic acid 4-[(5-nitro-2-pyridinyl)dithio]-2,5-dioxo-l - pyrrolidinyl ester (nitro-SPDB), 4-(Pyridin-2-yldisulfanyl)-2-sulfo-butync acid (sulfo- SPDB), N- succinimidyl (2 -pyridyldithio) propionate (SPDP), succinimidyl (N- maleimidomethyl) cyclohexane- 1 -carboxylate (SMCC), iminothiolane (IT),

[0234]

[0235] Docket No. 14887-WO

[0236] bifunctional derivatives of imidoesters (such as dimethyl adipimidate HCL), active esters (such as disuccinimidyl suberate), aldehydes (such as glutaraldehyde), bis-azido compounds (such as bis (p-azidobenzoyl)-hexanediamine), bis-diazonium derivatives (such as bis-(p-diazoniumbenzoyl)-ethylenediamine), diisocyanates (such as toluene 2,6-diisocyanate), and bis-active fluorine compounds (such as 1 ,5-difluoro-2,4- dinitrobenzene). For example, a ricin immunotoxin can be prepared as described in Vitettaet al (1987). Carbon labeled 1-isothiocyanatobenzyl methyldiethylene triaminepentaacetic acid (MX-DTPA) is an exemplary’ chelating agent for conjugation of radionucleotide to an antibody (WO 94 / 11026).

[0237]

[0128] In certain aspects, the linker is a "cleavable linker," which may facilitate release of the cytotoxic drug or other growth inhibitory’ agent inside of or in the vicinity of a cell, e.g., a tumor cell. In some aspects, the linker is a linker cleavable in an endosome of a mammalian cell. For example, an acid-labile linker, a peptidase-sensitive linker, an esterase labile linker, a photolabile linker or a disulfide-containing linker (see e.g., U.S.

[0238] Patent No. 5,208,020) may be used.

[0239]

[0129] The term “cytotoxic moiety ’' or sometimes “pay load,” refer to a chemical or biochemical moiety that is conjugated to the anti-CEACAM5 antibody described herein via a linker.

[0240]

[0130] In some aspects, the cytotoxic moiety is a camptothecin derivative, e.g., exatecan.

[0241]

[0131] The term “nucleic acid molecule,” as used herein, is used with reference to DNA molecules and RNA molecules. A nucleic acid molecule may be single-stranded or double-stranded, and may be a cDNA.

[0242]

[0132] The term “isolated nucleic acid molecule,” as used herein in reference to nucleic acids encoding antibodies or antibody fragments (e.g., Vn, VL, CDR3), is intended to refer to a nucleic acid molecule in which the nucleotide sequences are essentially free of other genomic nucleotide sequences, e.g, those encoding antibodies that bind antigens other than CEACAM5, yvhich other sequences may naturally flank the nucleic acid in human genomic DNA.

[0243]

[0133] The term “vector,” as used herein, is intended to refer to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. One type of vector is a “plasmid,” which refers to a circular double stranded DNA loop into which additional DNA segments may be ligated. Another type of vector is a viral vector, wherein additional DNA segments may be ligated into the viral genome. Certain

[0244]

[0245] Docket No. 14887-WO

[0246] vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors). Other vectors (e.g, non-episomal mammalian vectors) can be integrated into the genome of a host cell upon introduction into the host cell, and thereby are replicated along with the host genome. Moreover, certain vectors are capable of directing the expression of genes to which they are operatively linked. Such vectors are referred to herein as “recombinant expression vectors” (or simply, “expression vectors”). In general, expression vectors of utility in recombinant DNA techniques are often in the form of plasmids. In the present specification, “plasmid” and “vector” may be used interchangeably as the plasmid is the most commonly used form of vector. However, also included are other forms of expression vectors, such as viral vectors (e.g, replication defective retroviruses, adenoviruses and adeno-associated viruses), which serve equivalent functions.

[0247]

[0134] Also provided are “conservative sequence modifications” of the sequences set forth herein, e.g., amino acid sequence modifications which do not abrogate the binding of the antibody encoded by the nucleotide sequence or containing the amino acid sequence, to the antigen. Such conservative sequence modifications include conservative nucleotide and amino acid substitutions, as well as nucleotide and amino acid additions and deletions. For example, modifications can be introduced into a sequence by standard techniques known in the art, such as site-directed mutagenesis and PCR-mediated mutagenesis. Conservative amino acid substitutions include ones in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g, aspartic acid, glutamic acid), uncharged polar side chains (e.g, glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), nonpolar side chains (e.g, alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), beta-branched side chains (e.g, threonine, valine, isoleucine) and aromatic side chains (e.g, tyrosine, phenylalanine, tryptophan, histidine). Thus, a predicted nonessential amino acid residue in an anti- CEACAM5 antibody is preferably replaced with another ammo acid residue from the same side chain family. Methods of identifying nucleotide and amino acid conservative substitutions which do not eliminate antigen binding are well-known in the art (see, e.g.,

[0248]

[0249] Docket No. 14887-WO

[0250] Brummell et al., Biochem. 32:1180-1187 (1993); Kobayashi et al. Protein Eng.

[0251] 12(10):879-884 (1999); and Burks et al. Proc. Natl. Acad. Sci. USA 94:412-417 (1997)). Alternatively, in another aspect, mutations can be introduced randomly along all or part of an anti-CEACAM5 antibody coding sequence, such as by saturation mutagenesis, and the resulting modified anti-CEACAM5 antibodies can be screened for binding activity.

[0252]

[0135] For nucleic acids, the term “substantial homology'’ indicates that two nucleic acids, or designated sequences thereof, when optimally aligned and compared, are identical, with appropriate nucleotide insertions or deletions, in at least about 80% of the nucleotides, usually at least about 80% to 85%, 85% to 90% or 90% to 95%, and more preferably at least about 98% to 99.5% of the nucleotides. Alternatively, substantial homology7exists when the segments will hybridize under selective hybridization conditions, to the complement of the strand. For polypeptides, the term “substantial homology” indicates that two polypeptides, or designated sequences thereof, when optimally aligned and compared, are identical, with appropriate amino acid insertions or deletions, in at least about 80% of the amino acids, usually at least about 80% to 85%, 85% to 90%, 90% to 95%, and more preferably at least about 98% to 99.5% of the amino acids.

[0253]

[0136] The percent identity between two sequences is a function of the number of identical positions shared by the sequences (z.e., % homology = # of identical positions / total # of positions x 100), considering the number of gaps, and the length of each gap, which need to be introduced for optimal alignment of the two sequences. The comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm, as described in the non-limiting examples below.

[0254]

[0137] The percent identity7between two nucleotide sequences can be determined using the GAP program in the GCG software package (available at http: / / www.gcg.com), using a NWSgapdna.CMP matrix and a gap weight of 40, 50, 60, 70, or 80 and a length weight of 1, 2, 3, 4, 5, or 6. The percent identity between two nucleotide or two amino acid sequences can also be determined using the algorithm of E. Meyers and W. Miller (CABIOS, 4:11-17 (1989)) which has been incorporated into the ALIGN program (version 2.0), using a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty7of 4. In addition, the percent identity between two amino acid sequences can be determined using the Needleman and Wunsch (J. Mol. Biol. (48):444-453

[0255]

[0256] Docket No. 14887-WO

[0257] (1970)) algorithm which has been incorporated into the GAP program in the GCG software package (available at www.gcg.com), using either a Blossum 62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a length weight of 1, 2. 3, 4, 5. or 6.

[0258]

[0138] The nucleic acid and protein sequences described herein can further be used as a “query sequence” to perform a search against public databases to, for example, identify related sequences. Such searches can be performed using the NBLAST and XBLAST programs (version 2.0) of Altschul, et al. (1990) J. Mol. Biol. 215:403-10. BLAST nucleotide searches can be performed with the NBLAST program, score = 100, wordlength = 12 to obtain nucleotide sequences homologous to the nucleic acid molecules described herein. BLAST protein searches can be performed with the XBLAST program, score = 50, wordlength = 3 to obtain amino acid sequences homologous to the protein molecules described herein. To obtain gapped alignments for comparison purposes, Gapped BLAST can be utilized as described in Altschul et al., (1997) Nucleic Acids Res. 25(17):3389-3402. When utilizing BLAST and Gapped BLAST programs, the default parameters of the respective programs (e.g., XBLAST and NBLAST) can be used. See www.ncbi.nlm.nih.gov.

[0259]

[0139] The term “recombinant host cell” (or simply “host cell”), as used herein, is intended to refer to a cell that comprises a nucleic acid that is not naturally present in the cell and may be a cell into which a recombinant expression vector has been introduced. It should be understood that such terms are intended to refer not only to the particular subject cell but also to the progeny of such a cell. Because certain modifications may occur in succeeding generations due to either mutation or environmental influences, such progeny may not, in fact, be identical to the parent cell, but are still included within the scope of the term “host cell” as used herein.

[0260]

[0140] The term “inhibition,” as used herein, refers to any statistically significant decrease in biological activity, including partial and full blocking of the activity. For example, “inhibition” can refer to a statistically significant decrease of about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% in biological activity.

[0261]

[0141] The term “immunotherapy,” as used herein, refers to the treatment of a subject afflicted with, or at risk of contracting or suffering a recurrence of, a disease by a

[0262]

[0263] Docket No. 14887-WO

[0264] method comprising inducing, enhancing, suppressing or otherwise modifying an immune response.

[0265]

[0142] The terms “immunostimulating therapy” and “immunostimulatory therapy,” as used herein, refer to a therapy that results in an increase (e.g., inducing or enhancing) an immune response in a subject for, e.g., treating cancer.

[0266]

[0143] As used herein, “immune cell” refers to the subset of blood cells known as white blood cells, which include mononuclear cells such as lymphocytes, monocytes, macrophages, and granulocytes.

[0267]

[0144] As used herein, “abnormal” is used in the context of the activity or level or expression of a molecule which is outside of the normal activity or expression level ( g overexpressed) as compared to e.g., a control sample or reference sample exhibiting a normal activity / expression profile. The term “normal” is used herein in the context of the activity or level of expression of a protein found in a population of healthy, gender- and age-matched subjects. The minimal size of this healthy population may be determined using standard statistical measures, e.g., the practitioner could consider the incidence of the disease in the general population and the level of statistical certainty desired in the results. Preferably, the normal range of activity, level or expression of a biomarker is determined from a population of subjects (e.g., at least five, ten or twenty subjects), more preferably from a population of at least forty or eighty subjects, and even more preferably from more than 100 subjects.

[0268]

[0145] “T effector” (“Teff”) cells refer to T cells (e.g, CD4+ and CD8+ T cells) with cytolytic activities as well as T helper (Th) cells, which secrete inflammatory cytokines and activate and direct other immune cells but does not include regulatory T cells (Treg cells).

[0269]

[0146] As used herein, "administering" refers to the physical introduction of an anti- CEACAM5 ADC alone or in combination with another therapeutic agent (bevacizumab) to a subject, using any of the various methods and delivery systems known to those skilled in the art. Preferred routes of administration for ADCs described herein alone or in combination with bevacizumab include intravenous, intraperitoneal, intramuscular, subcutaneous, spinal or other parenteral routes of administration, for example by injection or infusion. The phrase "parenteral administration" as used herein means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intraperitoneal, intramuscular,

[0270]

[0271] Docket No. 14887-WO

[0272] intraarterial, intrathecal, intra-lymphatic, intralesional, intracapsular, intra-orbital, intracardiac, intradermal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural and intrastemal injection and infusion, as well as in vivo electroporation. Alternatively, an antibody described herein can be administered via a non-parenteral route, such as a topical, epidermal or mucosal route of administration, for example, intranasally, orally, vaginally, rectally, sublingually or topically. Administration can also be performed, for example, once, a plurality of times, and / or over one or more extended periods.

[0273]

[0147] As used herein, "cancer" refers to a broad group of diseases characterized by the uncontrolled growth of abnormal cells in the body. Unregulated cell division may result in the formation of malignant tumors or cells that invade neighboring tissues and may metastasize to distant parts of the body through the lymphatic system or bloodstream, and includes a variety of cancers, including but not limited to e.g., carcinomas, melanomas, sarcomas, leukemias, lymphomas, germ cell tumors, and blastomas.

[0274] Exemplary cancers for treatment include cancers of the brain, bladder, breast, cervix, colon, head and neck, kidney, lung, non-small cell lung, mesothelioma, ovary', prostate, stomach and uterus, leukemia, and medulloblastoma.

[0275]

[0148] As used herein, the term '‘small molecule drug” refers to a molecular entity, often organic or organometallic, that is not a polymer, that has medicinal activity, and that has a molecular weight less than about 2 kilodaltons (kDa), less than about 1 kDa, less than about 900 daltons (Da), less than about 800Da or less than about 700Da. The term encompasses most medicinal compounds termed “drugs” other than protein or nucleic acids, although a small peptide or nucleic acid analog can be considered a small molecule drug. Examples include chemotherapeutic anticancer drugs and enzy matic inhibitors. Small molecule drugs can be derived synthetically, semi-synthetically (i.e. , from naturally occurring precursors), or biologically.

[0276]

[0149] The terms “treat,” “treating,” and “treatment,” as used herein, refer to any type of intervention or process performed on, or administering an active agent (e.g., ADC comprising an anti-CEACAM5 antibody or antigen binding portion thereof described herein linked to a cytotoxic moiety via a linker as described herein) to, the subject with the objective of preventing, reversing, alleviating, ameliorating, inhibiting, or slowing down or preventing the progression, development, severity or recurrence of a symptom, complication, condition or biochemical indicia associated with a disease. Treatment

[0277]

[0278] Docket No. 14887-WO

[0279] can be of a subject having a disease or a subject who does not have a disease (e.g., for prophylaxis).

[0280]

[0150] As used herein, “adjunctive’' or “combined’' administration (co-administration) includes administration of an anti-CEACAM5 ADC (comprising an anti-CEACAM5 antibody or antigen binding portion thereof described herein linked to a cytotoxic moiety described herein via a linker) and bevacizumab in the same or different dosage form, or combined administration in separate dosages concurrently or sequentially. Thus, an anti-CEACAM5 ADC and bevacizumab can be simultaneously administered in a single formulation or formulated for separate administration and are administered concurrently or sequentially.

[0281]

[0151] ‘ ‘Combination” therapy, as used herein, means administration of two or more therapeutic agents in a coordinated fashion, and includes, but is not limited to, concurrent and sequential dosing. Specifically, combination therapy encompasses both co-administration (e.g., administration of a co-formulation or simultaneous administration of separate therapeutic compositions) and serial or sequential administration, provided that administration of one therapeutic agent is conditioned in some way on administration of another therapeutic agent. For example, one therapeutic agent may be administered only after a different therapeutic agent has been administered and allowed to act for a prescribed period of time. (See, e.g., Kohrt et al. (2011) Blood 117:2423). For example, the ADC (comprising an anti-CEACAM5 antibody linked to a cytotoxic moiety described herein via a linker) can be administered first followed by (e.g, immediately followed by) the administration of bevacizumab. or vice versa. In one aspect, the ADC is administered prior to administration of bevacizumab. In another aspect, the ADC is administered, for example, a few minutes (e.g., within about 30 minutes) or at least one hour of the bevacizumab. Such concurrent or sequential administration preferably results in both the ADC and bevacizumab being simultaneously present in treated patients.

[0282]

[0152] The administration of effective amounts of the ADC (comprising an anti-CEACAM5 antibody or antigen binding portion thereof described herein linked to a cytotoxic moiety described herein via a linker) alone, or the ADC combined with bevacizumab, according to any of the methods provided herein, can result in at least one therapeutic effect, including, for example, reduced tumor growth or size, reduced number of indicia of cancer (e.g., metastatic lesions) appearing over time, complete remission, partial

[0283]

[0284] Docket No. 14887-WO

[0285] remission, or stable disease. For example, the methods of treatment may produce a comparable clinical benefit rate (CBR = complete remission (CR)+ partial remission (PR) + stable disease (SD) lasting > 6 months) better than that achieved without administration of the ADC, or than that achieved with administration of any one of the ADC and the second agent, e.g., the improvement of clinical benefit rate is about 20% 20%, 30%, 40%, 50%, 60%, 70%, 80% or more.

[0286]

[0153] As used herein, the terms "inhibit" and "block" {e.g, with regard to inhibition / blocking of CEACAM5 binding or functional activity) are used interchangeably and encompass both partial and complete inhibition / blocking by the anti-CEACAM5 antibody or fragment thereof in the ADC, or other inhibition / blocking of a functional activity by a therapeutic agent. The degree of inhibition may be at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 100% (i.e., 2-fold or 2x), 3-fold, 5-fold or 10-fold relative to a control antibody or reference antibody. Additionally, the degree of inhibition may be between 20%-95%, 20%-80%, 20%-50%, 40%-95%, 40%- 80%, 40%-60%, 50%-90%, 50%-70%, 75%-95%, 75%-85%, 2-fold to 20-fold, 2-fold to 10-fold, 2-fold to 5-fold, 4-fold to 12-fold, or 4-fold to 8-fold.

[0287]

[0154] The term “effective dose” or “effective dosage” is defined as an amount sufficient to achieve or at least partially achieve a desired effect. A "therapeutically effective amount" or "therapeutically effective dosage" of a drug (e.g., ADC (comprising anti- CEACAM5 antibody or antigen binding portion thereof described herein linked to a cytotoxic moiety described herein via a linker)) is any amount of the drug or therapeutic agent that, when used alone or in combination with another therapeutic agent, promotes disease regression evidenced by a decrease in severity of disease symptoms, an increase or therapeutic agent in frequency and duration of disease symptom-free periods, or a prevention of impairment or disability due to the disease affliction. A therapeutically effective amount or dosage of a drug or therapeutic agent includes a " prophy lactically effective amount" or a "prophy lactically effective dosage", which is any amount of the drug or therapeutic agent that, when administered alone or in combination with another therapeutic agent to a subject at risk of developing a disease or of suffering a recurrence of disease, inhibits the development or recurrence of the disease. The ability of a therapeutic agent to promote disease regression or inhibit the development or recurrence of the disease can be evaluated using a variety of methods known to the skilled practitioner, such as in human subjects during clinical trials, in animal model systems

[0288]

[0289] Docket No. 14887-WO

[0290] predictive of efficacy in humans, or by assaying the activity of the agent in in vitro assays.

[0291]

[0155] By way of example, for the treatment of solid tumors, a therapeutically effective amount or dosage of the drug or therapeutic agent (e.g., ADC (comprising anti- CEACAM5 antibody or antigen binding portion thereof described herein linked to a cytotoxic moiety described herein via a linker), alone or optionally in combination with bevacizumab, inhibits tumor cell growth by at least about 20%, by at least about 30% by at least about 40%, by at least about 50%, by at least about 60%. by at least above 70%, by at least about 80%, or by at least about 90% relative to untreated subjects. In some aspects, a therapeutically effective amount or dosage of the drug or therapeutic agent completely inhibits cell grow th or tumor grow th, i. e. , inhibits cell growth or tumor growth by 100%. The ability of a compound or therapeutic agent, including an antibody, to inhibit tumor growth can be evaluated using the assays described herein. Alternatively, this property of a composition comprising the compound or therapeutic agent can be evaluated by examining the ability of the composition to inhibit cell growth; such inhibition can be measured in vitro by assays known to the skilled practitioner.

[0292]

[0156] The term '‘patient” includes human and other mammalian subjects that receive either prophylactic or therapeutic treatment.

[0293]

[0157] As used herein, the term “subject” includes any human or non-human animal. For example, the methods and compositions described herein can be used to treat a subject having cancer. The term “non-human animal” includes all vertebrates, e.g., mammals and non-mammals, such as non-human primates, sheep, cats, dogs, cows, chickens, amphibians, and reptiles.

[0294]

[0158] The term “sample” refers to tissue, bodily fluid, or a cell (or a fraction of any of the foregoing) taken from a patient or a subject. Normally, the tissue or cell will be removed from the patient, but in vivo diagnosis is also contemplated. In the case of a solid tumor, a tissue sample can be taken from a surgically removed tumor and prepared for testing. In certain embodiments, a core needle biopsy is performed using a hollow7needle to remove tissue samples from the cancer. In the case of lymphomas and leukemias, lymphocytes, leukemic cells, or lymph tissues can be obtained (e.g., leukemic cells from blood) and appropriately prepared. Other samples, including e.g,

[0295]

[0296] Docket No. 14887-WO

[0297] urine, tears, serum, plasma, cerebrospinal fluid, feces, sputum, and cell extracts can also be useful for particular cancers.

[0298]

[0159] The terms “detection” or “detected”, as used herein refer to qualitative and / or quantitative detection (measuring levels) with or without reference to a control.

[0299]

[0160] The term “diagnosing”, as used herein, means the determination of the nature of a medical condition intended to identify a pathology' which affects the subject from a number of collected data.

[0300]

[0161] As used herein, "comprising" is synonymous with "including," "containing," "having" or "characterized by," and is inclusive or open-ended and does not exclude additional, unrecited elements or method steps. As used herein, "consisting of excludes any element, step, or ingredient not specified in the claim element. As used herein, "consisting essentially of does not exclude materials or steps that do not materially affect the basic and novel characteristics of the claim. In each instance herein any of the terms "comprising," "consisting essentially of," and "consisting of may be optionally replaced with either of the other two terms, thus describing alternative aspects of the scope of the subject matter. The invention illustratively described herein suitably may¬ be practiced in the absence of any element or elements, limitation or limitations which is not specifically disclosed herein.

[0301]

[0162]

[0302]

[0163] As used herein, the term "stereoisomer" refers to isomers of identical constitution that differ in the arrangement of their atoms in space. Enantiomers and diastereomers are examples of stereoisomers. Geometric isomers are also examples of stereoisomers. The term "enantiomer" refers to one of a pair of molecular species that are mirror images of each other and are not superimposable. The term "diastereomer" refers to stereoisomers that are not mirror images. The term "racemate" or "racemic mixture" refers to a composition composed of equimolar quantities of two enantiomeric species, wherein the composition is devoid of optical activity. Geometric isomers of C=C double bonds can also be present in the ADCs, and all such stable isomers are contemplated in the present invention. Cis- and trans- (or E- and Z-) geometric isomers of the ADCs of the present invention are described and may be isolated as a mixture of isomers or as separated isomeric forms.

[0303]

[0164] Various aspects described herein are described in further detail in the following subsections.

[0304]

[0305] Docket No. 14887-WO

[0306]

[0165] The present disclosure provides an antibody drug conjugate comprising an anti- CEACAM5 antibody or antigen binding portion thereof described herein which is linked or conjugated via a phosphorus (V) moiety (also denoted as “P5"’) and a linker to a cytotoxic moiety, i.e., camptothecin or derivatives and analogs thereof.

[0307]

[0166] In some aspects, the present disclosure provides a method of preparing an ADC of the formula (I), comprising reacting Compound A with a thiol containing compound, AB- (SH)n, wherein AB is the anti-CEACAM5 antibody or antigen binding portion thereof disclosed herein, to obtain the ADC of the formula (I):

[0308]

[0309] (I), or a pharmaceutically acceptable salt or solvate thereof, wherein n is as defined above. Methods of selective bioconjugation reaction of ethynylphosphonami deates with cysteine containing compounds have been described in WO2018041985A1 (published March 8, 2018), W02019170710A2 (published September 12, 2019), WO2022223783A1 (published October 27, 2022), W02023083900A1 (published May- 19, 2023), WO2023083919A1 (published May 19, 2023), each of which is incorporated herein by reference.

[0310]

[0168] The number of cytotoxic moieties linked to the antigen binding moiety of a CEACAM5- ADC (drug-to-antibody ratio: DAR) can vary and will be limited only by the number of available attachments sites on the antigen binding moiety and the number of agents linked to a single linker.

[0311]

[0169] The DAR value can vary with the nature of the antigen binding moiety (e.g., any antibody or the antigen-binding portion thereof described herein) and the drug used along with the experimental conditions used for the conjugation (DAR, reaction time, nature of the solvents and / or cosolvents). Thus, the contact between the antibody and the drug may in an ADC lead to a mixture comprising several conjugates differing from one another by different drug-to-antibody ratios and may further include free antibodies

[0312]

[0313] Docket No. 14887-WO

[0314] and / or aggregates. The DAR that is determined is thus a mean value. DARs may be analyzed by UV spectrometry, monomer content may be analyzed by SEC-HPLC, and free drug content may be analyzed by RP-HPLC.

[0315]

[0170] In some aspects, a linker will link a single cytotoxic moiety to the antigen binding moiety (e.g.. any antibody or the antigen-binding portion thereof described herein) of a conjugate. In some aspects where the conjugate includes more than one cytotoxic moiety, each moiety7may be the same or different. As long as the conjugate does not exhibit unacceptable levels of aggregation under the conditions of use and / or storage, conjugates with DARs of twenty, or even higher, are contemplated. In some aspects, the conjugates described herein may have a DAR in the range of about 1-10, 2-10, 1-8, 2-8, 1-6, 2-6, 1-4, or 2-4. In some specific aspects, the conjugate may have a DAR of 2, 3. 4 or 5. In some aspects, the DAR is 6. In some aspects, the DAR is 7. In some aspects, the DAR is 8. In some aspects, the DAR is 9. In some aspects, the DAR is 6 or 7. In some aspects, the DAR is 7, 7.5, or 8. In some aspects, the DAR is 7-8.

[0316]

[0171] In some aspects, an ADC of the present disclosure has the following structure:

[0317]

[0318]

[0173] or a pharmaceutically acceptable salt thereof, wherein represents that the configuration of the double bond may be E or Z; AB is the antibody, or antigen binding portion thereof that binds CEACAM5. comprising a VH comprising CDR1, CDR2, and CDR3 regions comprising the amino acid sequences set forth in SEQ ID NOs: 1, 2, and 3, respectively, and a VL comprising CDR1, CDR2, and CDR3 regions comprising the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10; a VH and a VL. In certain embodiments, the AB comprises a VH and VL which comprise the amino acid sequences set forth in SEQ ID NOs: 4 and 11, respectively. In certain embodiments, the AB comprises a heavy chain and a light chain which comprise the amino acid sequence

[0319]

[0320] Docket No. 14887-WO

[0321] set forth in SEQ ID NO: 6 and the amino acid sequence set forth in SEQ ID NO: 13, respectively.

[0322]

[0174] The anti-CEAC AM5 antibodies useful for the ADC of the disclosure can be defined by particular structural features.

[0323]

[0175] As used herein, the terms “carcinoembryonic antigen-related cell adhesion molecules’’ and "CEACAM5" are used interchangeably with reference to human CEACAM5 or cynomolgus (Macaca fascicular is) CEACAM5, unless the context clearly dictates otherwise. The human CEACAM5 precursor polypeptide (with signal peptide) contains the amino acid sequence set forth in the SEQ ID NO: 15 (GenBank: AAH34671.1), which is shown below.

[0324]

[0325]

[0176] The cynomolgus CEACAM5 precursor polypeptide (with signal peptide) contains the amino acid sequence set forth in SEQ ID NO: 16 (NCBI: XM 005589434.3). The amino acid sequence and nucleic acid sequences of human CEACAM5 and cynomolgus CEACAM5 are shown below.

[0326]

[0327] Docket No. 14887-WO

[0328]

[0329]

[0177] The term “CEACAM5” further includes counterparts from other species and other naturally occurring allelic, splice variants, and processed forms thereof, unless the context clearly dictates otherwise.

[0330]

[0178] In some aspects, the isolated anti-CEACAM5 antibody (e.g, recombinant humanized, chimeric, or human antibody) or antigen binding portion thereof (e.g., useful for conjugation to a cytotoxic moiety described herein to produce an ADC and useful for being the antibody in the ADC) is found in Table 3. The anti-CEACAM5 antibody or antigen binding portion thereof binds to and internalizes into CEACAM5-expressing cells. Thus, the anti-CEACAM5 antibody or antigen binding portion is useful for the ADC (comprising the antibody or antigen binding portion thereof linked to a cytotoxic moiety) by effectively delivering the cytotoxic moiety to kill cells, for example cancer cells.

[0331]

[0179] Anti-CEACAM5 antibodies useful for the ADC include all known forms of antibodies and other protein scaffolds with antibody-like properties. For example, the antibody can be a monoclonal antibody, a humanized antibody, a human antibody, a bispecific antibody, an immunoconjugate, a chimeric antibody, or a protein scaffold with antibody-like properties, such as fibronectin or ankyrin repeats. The antibody also can be a Fab, F(ab’)2, scFv, AFFIBODY, avimer, nanobody, single chain antibody, or a domain antibody. The antibody also can have any isotype or allotype, including any of the following isotypes: IgGl, IgG2, IgG3, IgG4, IgM, IgAl, IgA2, secretory IgA (SIgA). IgD, IgE, and allotypes thereof. Full-length antibodies can be prepared from VH and Vi. sequences using standard recombinant DNA techniques and nucleic acid encoding the desired constant region sequences to be operatively linked to the variable region sequences.

[0332]

[0333] Docket No. 14887-WO

[0334]

[0180] In an embodiment of the method, the antibody, or antigen binding portion thereof, comprises a bispecific molecule comprising the anti-CEACAM5 antibody or antigen binding portion thereof and a second binding region that binds to another antigen.

[0335]

[0181] In an embodiment of the method, the antibody, or antigen binding portion thereof, comprises a multispecific molecule comprising the anti-CEACAM5 antibody, or antigen binding portion thereof and at least two binding regions, each of which binds other antigens.

[0336]

[0182] In some aspects, the anti-CEACAM5 antibody, or antigen binding portion thereof, which binds human CEACAM5 and is useful for conjugation to the cytotoxic moiety for producing the ADC has at least one amino acid mutation compared to an amino acid sequence described in Table 3.

[0337]

[0183] In some aspects, the anti-CEACAM5 antibody, or antigen binding portion thereof, useful for the ADC specifically binds to CEACAM5 with a KD less than 1 X 10‘6M. In some aspects, the anti-CEACAM5 antibody, or antigen binding portion thereof, useful for the ADC specifically binds to CEACAM5 with a KD less than 1 X 10'7M. In some aspects, the anti-CEAC AM5 antibody, or antigen binding portion thereof, useful for the ADC specifically binds to CEACAM5 with a D less than 1 X 10’8M. In some aspects, the anti-CEACAM5 antibody, or antigen binding portion thereof, useful for the ADC specifically binds to CEACAM5 with a KD less than 5 X 10'9M. In some aspects, the anti-CEACAM5 antibody, or antigen binding portion thereof, useful for the ADC specifically binds to CEACAM5 with a KD less than 1 X 10'9M. In some aspects, the anti-CEACAM5 antibody, or antigen binding portion thereof, useful for the ADC specifically binds to CEACAM5 with aKn less than 5 X 10‘10M.

[0338]

[0184] Antibody engineering of variable regions

[0339]

[0185] In some aspects, the ADC or its components (e.g, anti-CEACAM5 antibody or antigen binding portion thereof) are engineered with modifications to framework residues within the variable domains of the parental antibody, e.g.. to improve the properties of the antibody or antigen binding portion thereof. Typically, such framework modifications are made to decrease the immunogenicity of the anti- CEACAM5 antibodies or antigen binding portions thereof. This is usually accomplished by replacing non-CDR residues in the variable domains (i.e.. framework residues) in a parental (e.g., rodent) antibody with analogous residues from the immune repertoire of the species in which the antibody is to be used, e.g., human residues in the

[0340]

[0341] Docket No. 14887-WO

[0342] case of human therapeutics. Such an antibody is referred to as a "humanized" antibody. In some cases, it is desirable to increase the affinity, or alter the specificity of an engineered (e.g., humanized) antibody. One approach is to "back-mutate" one or more framework residues to the corresponding germline sequence. More specifically, an antibody that has undergone somatic mutation can contain framework residues that differ from the germline sequence from which the antibody is derived. Such residues can be identified by comparing the antibody framework sequences to the germline sequences from which the antibody is derived. Another approach is to revert to the original parental (e.g, rodent) residue at one or more positions of the engineered (e.g, humanized) antibody, e.g., to restore binding affinity that may have been lost in the process of replacing the framework residues. (See, e.g, U.S. Patent No. 5,693,762, U.S. Patent No. 5,585,089 and U.S. Patent No. 5,530,101.)

[0343]

[0186] In certain aspects, the anti-CEACAM5 antibodies and antigen binding portions thereof in an ADC are engineered (e.g. humanized) to include modifications in the framework and / or CDRs to improve their properties. Such engineered changes can be based on molecular modeling. A molecular model for the variable region for the parental (non-human) antibody sequence can be constructed to understand the structural features of the antibody and used to identify potential regions on the antibody that can interact with the antigen. Conventional CDRs are based on alignment of immunoglobulin sequences and identifying variable regions. Kabat et al., (1991) Sequences of Proteins of Immunological Interest, Kabat, et al , National Institutes of Health. Bethesda, Md. ; 5thed.; NIH Publ. No. 91-3242: Kabat (1978) Adv. Prot. Chem.

[0344] 32:1-75; Kabat, et al., (1977) .7 Biol. Chem. 252:6609-6616. Chothia and coworkers carefully examined conformations of the loops in crystal structures of antibodies and proposed hypervariable loops. Chothia, et al., (1987) J Mol. Biol. 196:901-917 or Chothia, et al., (1989) Nature 342:878-883. There are variations between regions classified as “CDRs” and “hypervariable loops”. Later studies (Raghunathan et al., (2012) J. Mol Recog. 25, 3, 103-113) analyzed several antibody-antigen crystal complexes and observed that the antigen binding regions in antibodies do not necessarily conform strictly to the “CDR” residues or “hypervariable” loops. The molecular model for the variable region of the non-human antibody can be used to guide the selection of regions that can potentially bind to the antigen. In practice, the potential antigen binding regions based on model differ from the conventional “CDR”s

[0345]

[0346] Docket No. 14887-WO

[0347] or “hyper variable” loops. Commercial scientific software such as MOE (Chemical Computing Group) can be used for molecular modeling. Human frameworks can be selected based on best matches with the non-human sequence both in the frameworks and in the CDRs. For FR4 (framework 4) in VH, VJ regions for the human germlines are compared with the corresponding non-human region. In the case of FR4 (framework 4) in VL, J-kappa and J-Lambda regions of human germline sequences are compared with the corresponding non-human region. Once suitable human frameworks are identified, the CDRs are grafted into the selected human frameworks. In some cases, certain residues in the VL-VH interface can be retained as in the non-human (parental) sequence. Molecular models can also be used for identifying residues that can potentially alter the CDR conformations and hence binding to antigen. In some cases, these residues are retained as in the non-human (parental) sequence. Molecular models can also be used to identify solvent exposed amino acids that can result in unwanted effects such as glycosylation, deamidation and oxidation. Developability filters can be introduced early on in the design stage to eliminate / minimize these potential problems.

[0348]

[0187] Another type of framework modification involves mutating one or more residues within the framework region, or even within one or more CDR regions, to remove T cell epitopes to thereby reduce the potential immunogenicity of the antibody. This approach is also referred to as "deimmunization" and is described in further detail in U.S. Patent No. 7,125,689. In certain aspects, one or more glycosylation sites in either the light or heavy chain immunoglobulin variable regions, such as the framework regions, may be modified or removed to reduce immunogenicity. In particular aspects, it will be desirable to change certain amino acids containing exposed side-chains to another amino acid residue in order to provide for greater chemical stability of the final antibody, so as to avoid deamidation or isomerization. The deamidation of asparagine may occur on NG, DG, NG, NS. NA, NT, QG or QS sequences and result in the creation of an isoaspartic acid residue that introduces a kink into the polypeptide chain and decreases its stability (isoaspartic acid effect). Isomerization can occur at DG, DS, DA or DT sequences. In certain aspects, the antibodies provided herein do not contain deamidation or asparagine isomerism sites. For example, an asparagine (Asn) residue may be changed to Gin or Ala to reduce the potential for formation of isoaspartate at any Asn-Gly sequences, particularly within a CDR.

[0349]

[0350] Docket No. 14887-WO

[0351]

[0188] A similar problem may occur at an Asp-Gly sequence. Reissner and Aswad (2003) Cell. Mol. Life Sei. 60:1281. Isoaspartate formation may debilitate or completely abrogate binding of an antibody to its target antigen. See, Presta (2005) J. Allergy’ Clin. Immunol. 116:731 at 734.

[0352]

[0189] In various aspects, the asparagine is changed to glutamine (Gin). It may also be desirable to alter an amino acid adjacent to an asparagine (Asn) or glutamine (Gin) residue to reduce the likelihood of deamidation, which occurs at greater rates when small amino acids occur adjacent to asparagine or glutamine. See. Bischoff & Kolbe (1994) J. Chromatog. 662:261. In addition, any methionine residues (typically solvent exposed Met) in CDRs may be changed to Lys, Leu, Ala, or Phe or other amino acids in order to reduce the possibility that the methionine sulfur would oxidize, which could reduce antigen-binding affinity and also contribute to molecular heterogeneity in the final antibody preparation. Id. Additionally, in order to prevent or minimize potential scissile Asn-Pro peptide bonds, it may be desirable to alter any Asn-Pro combinations found in a CDR to Gin-Pro, Ala-Pro, or Asn- Ala. Antibodies with such substitutions are subsequently screened to ensure that the substitutions do not decrease the affinity or specificity of the antibody for CEACAM5, or other desired biological activity to unacceptable levels. See Table 1 for exemplary stabilizing CDR variants.

[0353]

[0190] Table 1. Exemplary stabilizing CDR variants

[0354]

[0355]

[0191] Pharmaceutical Compositions

[0356]

[0192] Also provided herein are pharmaceutical compositions comprising an ADC as disclosed herein and a carrier (e.g., pharmaceutically acceptable carrier). Such compositions are useful for various therapeutic applications, such as cancer treatment.

[0357]

[0358] Docket No. 14887-WO

[0359]

[0193] In some aspects, the pharmaceutical compositions may further include other compounds, drugs, and / or agents for various therapeutic applications. Such compounds, drugs, and / or agents can include, for example, an anti-cancer agent, a chemotherapeutic agent, an immunosuppressive agent, an immunostimulatory agent, an immune checkpoint inhibitor, and / or an anti-inflammatory agent. Exemplary compounds, drugs, and agents that can be formulated together or separately with the ADC described in the next section.

[0360]

[0194] As used herein, "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible. In some aspects, the carrier is suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal or epidermal administration (e.g., by injection or infusion). Depending on the route of administration, the active compound, i.e., antibody, immunoconjugate, or bispecific molecule, may be coated in a material to protect the compound from the action of acids and other natural conditions that may inactivate the compound.

[0361]

[0195] The pharmaceutical compounds described herein may include one or more pharmaceutically acceptable salts. A "pharmaceutically acceptable salt" refers to a salt that retains the desired biological activity of the parent compound and does not impart any undesired toxicological effects (see e.g., Berge, S.M., et al. (1977) J. Pharm. Sci.

[0362] 66: 1-19). Examples of such salts include acid addition salts and base addition salts. Acid addition salts include those derived from nontoxic inorganic acids, such as hydrochloric, nitric, phosphoric, sulfuric, hydrobromic, hydroiodic, phosphorous and the like, as well as from nontoxic organic acids such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxy alkanoic acids, aromatic acids, aliphatic and aromatic sulfonic acids and the like. Base addition salts include those derived from alkaline earth metals, such as sodium, potassium, magnesium, calcium and the like, as well as from nontoxic organic amines, such as N,N'- dibenzyl ethylenedi amine, N-methylglucamine, chloroprocaine, choline, diethanolamine, ethylenediamine, procaine and the like.

[0363]

[0196] A pharmaceutical composition described herein may also include a pharmaceutically acceptable antioxidant. Examples of pharmaceutically acceptable antioxidants include: (1) water soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; (2) oil-soluble antioxidants,

[0364]

[0365] Docket No. 14887-WO

[0366] such as ascorbyl palmitate, butylated hydroxyanisole (BHA), but lated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol, and the like; and (3) metal chelating agents, such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

[0367]

[0197] Examples of suitable aqueous and nonaqueous carriers that may be employed in the pharmaceutical compositions described herein include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate. These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of presence of microorganisms may be ensured both by sterilization procedures, supra, and by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents, such as sugars, sodium chloride, and the like into the compositions. In addition, prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents which delay absorption such as aluminum monostearate and gelatin.

[0368]

[0198] Pharmaceutically acceptable carriers include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. Except insofar as any media or agent is incompatible with the active compound, use thereof in the pharmaceutical compositions described herein is contemplated. A pharmaceutical composition may comprise a preservative or may be devoid of a preservative. Supplementary active compounds can be incorporated into the compositions.

[0369]

[0199] A composition described herein can be administered via one or more routes of administration using one or more of a variety of methods. The route and / or mode of administration can vary depending upon the desired results. Routes of administration for the ADC described herein include e.g., intravenous, intramuscular, intradermal, intraperitoneal, subcutaneous, spinal or other parenteral routes of administration, for example by injection or infusion. The phrase "parenteral administration" as used herein means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal,

[0370]

[0371] Docket No. 14887-WO

[0372] transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural and intrastemal injection and infusion.

[0373]

[0200] Alternatively, an ADC described herein can be administered via a non-parenteral route, such as a topical, epidermal or mucosal route of administration, for example, intranasally, orally, vaginally, rectally, sublingually or topically.

[0374]

[0201] Uses and Methods

[0375]

[0202] The ADC alone or in combination with bevacizumab described herein have numerous in vitro and in vivo utilities as described herein.

[0376]

[0203] In one aspect, provided herein is a method of treating cancer comprising administering to a subject in need thereof an ADC in a therapeutically effective dose or amount so that the growth of a cancerous tumor is inhibited or reduced and / or that regression and / or that prolonged survival is achieved.

[0377]

[0204] In some aspects, the ADC described herein may be administered in combination with additional cytotoxic or therapeutic agent(s), for example as described herein.

[0378]

[0205] Cancers that express CEACAM5 whose grow th may be inhibited using the ADC described herein include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia More particular examples of such cancers include, but are not limited to, basal cell carcinoma, biliary tract cancer; bladder cancer; bone cancer; brain and CNS cancer; breast cancer (e.g. estrogen-receptor positive breast cancer HER2- positive breast cancer; triple negative breast cancer); cancer of the peritoneum; cervical cancer; cholangiocarcinoma; choriocarcinoma; colon and rectum cancer; connective tissue cancer; cancer of the digestive system; endometrial cancer; esophageal cancer; eye cancer; cancer of the head and neck; gastric cancer (including gastrointestinal cancer); glioblastoma; liver cancer (e.g., hepatocellular carcinoma; hepatoma); intraepithelial neoplasm; kidney or renal cancer; larynx cancer; leukemia; lung cancer (e.g.. small-cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung, and squamous carcinoma of the lung); lymphoma including Hodgkin's and non-Hodgkin's lymphoma; melanoma; myeloma; neuroblastoma; oral cavity7cancer (e.g., lip, tongue, mouth, and pharynx); ovarian cancer; pancreatic cancer; prostate cancer; retinoblastoma; rhabdomyosarcoma; rectal cancer; cancer of the respiratory system; salivary7gland carcinoma; sarcoma; skin cancer; squamous cell cancer; teratocarcinoma; testicular cancer; thyroid cancer; uterine or endometrial cancer; cancer of the urinary7

[0379]

[0380] Docket No. 14887-WO

[0381] system; vulval cancer; as well as other carcinomas and sarcomas; as well as B-cell lymphoma (including low grade / fbllicular non-Hodgkin's lymphoma (NHL); small lymphocytic (SL) NHL; intermediate grade / fbllicular NHL; intermediate grade diffuse NHL; high grade immunoblastic NHL; high grade lymphoblastic NHL; high grade small non-cleaved cell NHL; bulky disease NHL; mantle cell lymphoma; AIDS-related lymphoma; and Waldenstrom's Macroglobulinemia); chronic lymphocytic leukemia (CLL); acute lymphoblastic leukemia (ALL); Hairy cell leukemia; chronic myeloblasts leukemia; and post-transplant lymphoproliferative disorder (PTLD), as well as abnormal vascular proliferation associated with phakomatoses, edema (such as that associated with brain tumors), tumors of primitive origins and Meigs' syndrome.

[0382]

[0206] Additional cancers which express CEACAM5 and can be treated using the ADC described herein include metastatic pancreatic cancer, metastatic adenocarcinoma of the pancreas, stomach cancer, fibrotic cancer, glioma, malignant glioma, diffuse intrinsic pontine glioma, recurrent childhood brain neoplasm renal cell carcinoma, clear-cell metastatic renal cell carcinoma, metastatic castration resistant prostate cancer, stage IV prostate cancer, metastatic melanoma, malignant melanoma, recurrent melanoma of the skin, melanoma brain metastases, malignant melanoma of head and neck, squamous cell non-small cell lung cancer, metastatic breast cancer, follicular lymphoma, advanced B- cell NHL, HL including diffuse large B-cell lymphoma (DLBCL), multiple myeloma, chronic myeloid leukemia, adult acute myeloid leukemia in remission, adult acute myeloid leukemia with Inv(16)(pl3. Iq22), CBFB-MYH11, adult acute myeloid leukemia with t(16: 16) (p 13.1 :q22), CBFB-MYH11, adult acute myeloid leukemia with t(8:21)(d22:q22), RUNX1-RUNX1T1, adult acute myeloid leukemia with t(9:ll)(p22:q23), MLLT3-MLL, adult acute promyelocytic leukemia with

[0383] tO15: 17)(q22:ql2), PML-RARA, alkylating agent-related acute myeloid leukemia, Richter's syndrome, adult glioblastoma, adult gliosarcoma, recurrent glioblastoma, recurrent childhood rhabdomyosarcoma, recurrent Ewing sarcoma / peripheral primitive neuroectodermal tumor, recurrent neuroblastoma, recurrent osteosarcoma, colorectal cancer, MSI positive colorectal cancer, MSI negative colorectal cancer, nasophary ngeal nonkeratinizing carcinoma, recurrent nasopharyngeal undifferentiated carcinoma, cervical adenocarcinoma, cervical adenosquamous carcinoma; cervical squamous cell carcinoma, recurrent cervical carcinoma, anal canal squamous cell carcinoma, metastatic anal canal carcinoma, recurrent anal canal carcinoma, recurrent head and

[0384]

[0385] Docket No. 14887-WO

[0386] neck cancer, squamous cell of head and neck, head and neck squamous cell carcinoma (HNSCC), ovarian carcinoma, colon cancer, advanced GI cancer, gastric adenocarcinoma, gastroesophageal junction adenocarcinoma, bone neoplasms, soft tissue sarcoma, bone sarcoma, thymic carcinoma, urothelial carcinoma. Merkel cell carcinoma, recurrent Merkel cell carcinoma, mycosis fungoides, Sezary syndrome, neuroendocrine cancer, nasopharyngeal cancer, basal cell skin cancer, squamous cell skin cancer, dermatofibrosarcoma trotuberans, glioma, mesothelioma, myelodysplastic syndromes (MDS). myelofibrosis (MF), myeloproliferative neoplasms, and acute myeloid leukemia (AML).

[0387]

[0207] Cancers may be, e.g., metastatic or primary cancers; desmoplastic or non- desmoplastic cancers; or recurrent cancers.

[0388]

[0208] In some aspects, the cancer is resistant to checkpoint inhibitor(s). In some aspects, the cancer is intrinsically refractory or resistant (e.g., resistant to a PD-1 pathway inhibitor, PD-1 pathway inhibitor, or CTLA-4 pathway inhibitor). In some aspects, the resistance or refractory state of the cancer is acquired. In some aspects, the ADC described herein can be used in combination with checkpoint inhibitors to overcome resistance of the cancer to the checkpoint inhibitors. In some aspects, the ADC described herein can be used to treat tumors with a mesenchymal and / or EMT signature together with checkpoint inhibitors in combination or sequentially with agents that induce a mesenchymal phenotype, such as MAPK pathway inhibitors.

[0389]

[0209] In some aspects, the ADCs described herein are used to enhance the viability of immune cells ex vivo, e.g.. in adoptive NK cell transfer. Accordingly, in some aspects. ADCs are used in combination with adoptively transferred NK cells to treat cancer. In some aspects, the ADC described herein are used to treat tumors with MHC loss or MHC down-regulation, as monotherapy or in combination with NK activating or enhancing treatment.

[0390]

[0210] Combination therapy

[0391] [2H] The ADCs described herein can be used in combination with bevacizumab or additional various treatments or agents known in the art for the treatment of disease or condition, as described herein.

[0392]

[0393] Docket No. 14887-WO

[0394]

[0212] In some aspects, a method of treating cancer comprises administering to a subject in need thereof a therapeutically effective amount of an ADC described herein in combination with bevacizumab.

[0395]

[0213] In some aspects, the ADC described herein may also be administered with a standard of care treatment, or another treatment, such as radiation, surgery, or chemotherapy.

[0396]

[0214] Kits

[0397]

[0215] Also provided are kits comprising an ADC described herein alone or in combination with bevacizumab and instructions for use.

[0398]

[0216] In some aspects, the kits comprise the ADC in unit dosage form, such as in a single dose vial or a single dose pre-loaded syringe, optionally contained in a single vial or container, along with e.g., instructions for use in treating a cancer as described herein.

[0399]

[0217] The present disclosure is further illustrated by the following examples, which should not be construed as further limiting. The contents of all figures and all references, Genbank sequences, issued patents, and published patent applications cited throughout this disclosure are expressly incorporated herein by reference.

[0400]

[0218] Table 2. Abbreviations used in Examples

[0401]

[0402]

[0403] Docket No. 14887-WO

[0404]

[0405]

[0406] Docket No. 14887-WO

[0407]

[0408]

[0409] Docket No. 14887-WO

[0410]

[0411]

[0412] Docket No. 14887-WO

[0413]

[0414]

[0415] Docket No. 14887-WO

[0416]

[0417]

[0418] Docket No. 14887-WO

[0419]

[0420] Docket No. 14887-WO

[0421]

[0422]

[0219] Example 1. Generation of ADC001

[0423]

[0220] This example describes the conjugation of antibody MAB1 antibody (amino acid and nucleic acid sequence listed in Table 3) to Compound A’ to generate ADC001. Table 4 lists the non-MABl materials used for the conjugation method.

[0424]

[0221] Table 3. Summary table of amino acid sequences and nucleic acid sequences for MAB1

[0425]

[0426] Docket No. 14887-WO

[0427]

[0428]

[0429] Docket No. 14887-WO

[0430]

[0431] Docket No. 14887-WO

[0432]

[0433] Table 4. Non-Mab conjugation reagents and consumables

[0434]

[0435]

[0436] (Compound A’)

[0437]

[0222] Firstly, each mAb was buffer-exchanged into P5 conjugation buffer (50 mM Tris, 100 mM NaCl, 1 mM EDTA, pH 8.3 at 25°C) and adjusted to a concentration of 10 mg / ml. Next. Zeba Spin desalting columns were utilized. The columns were equilibrated with P5 conjugation buffer according to the manufacturers' instructions. Recover}7-yields were usually > 95%.

[0438]

[0223] For the mAb stock solution with a concentration below 10 mg / ml, concentration and buffer exchange (diafiltration) were performed using protein concentrator spin columns (Amicon Ultra) according to the manufacturers' instructions. The spin columns are rinsed with P5 conjugation buffer prior to application of the mAb.

[0439]

[0224] After buffer exchange, the mAb concentration was measured with a nanophotometer.

[0440] P5 conjugation buffer was utilized as the blank. Finally, the mAb concentration was adjusted to 10 mg / ml with P5 conjugation buffer.

[0441]

[0225] Next the mAb was transferred into amber plasticware. The conjugation reaction was performed protected from light. Conjugation with Compound A’ was performed with a molar ratio of 7 eq. TCEP and 10 eq. of Compound A’ per 1 equivalent of MAB 1 mAb.

[0442]

[0443] Docket No. 14887-WO

[0444] Typically, a 40 mM Compound A’ stock in DMSO and a 10 mM TCEP working solution were used. The 40 mM linker-payload stock solution was thawed. A fresh 10 mM TCEP working solution was prepared by combining 20 pl of 0.5 M TECP-HC1 pH 7.0 with 980 pl of P5 conjugation buffer. Both reagents were vortexed prior to use.

[0445]

[0226] The calculated amount of TCEP working solution was added to the solution of the MAB1 mAb and mixed by gentle swirling. The calculated amount of Compound A’ stock was added immediately afterwards. The mixture was then incubated overnight at 23°C in amber 50 ml tubes and spun at a speed of 300 rpm.

[0446]

[0227] Conjugation efficiency was evaluated by LC-MS analysis. The conjugate samples were diluted to 1 mg / mL in 100 mM Tris pH 7.5. 20 pl of sample was reduced by adding 2 pl of 0.5M dithiothreitol (DTT) or TCEP. The samples were analyzed by LC- MS using an Agilent 1290 Infinity UPLC system coupled to a 6530 Accurate-Mass Q- TOF mass spectrometer. The analytical column (Waters Inc., BEH C4 column. 1.7 um, 2.1 mm x 50 mm) was equilibrated at 60°C. The mobile phase consisted of 0.1% formic acid in water (phase A) and 0.1% formic acid in acetonitrile (phase B). The system was operated at a flow rate of 200 pl / min. The gradient condition was as follows: 0-2 min., held at 27% B; 2-9 min., slow ramp from 27-37% B; 9-9.5 min., linear ramp from 37- 90% B; 9.5-12.3 min., held at 90% B. The MS settings were as follows: polarity = positive, capillary voltage = 4.2 kV, sample cone = 4d V, source offset = 15 V, source temperature = 140°C, desolvation temperature = 325°C. The data acquisition range was 900-3200 m / z. Deconvolution was done using Agilent MassHunter Walkup software. If any unconjugated mAb was still present, another 1.4 equivalents of TCEP and 2 equivalents of Compound A’ were added followed by incubation for 2-4 hours. The process yielded DAR8 ADCs.

[0447]

[0228] Example 2. Pre-clinical analysis of ADC001

[0448]

[0229] ADC001 was designed as a potentially best-in-class ADC comprised of a CEACAM5 targeting human IgGl conjugated to exatecan via the Tubutecan plasma-stable linker. ADC001 selectively binds CEACAM5 on the tumor cell surface and delivered the payload exatecan intracellularly to induce DNA-damage and antitumor activity alone or in combination with anti-VEGF (bevacizumab) in multiple murine CDX and PDX models. In heterogenous tumor settings, ADC001 induced cellular growth inhibition and tumor regressions via the secondary7mechanism of bystander kill effect imparted by

[0449]

[0450] Docket No. 14887-WO

[0451] the high cellular permeability of exatecan. The results of the nonclinical toxicology studies with ADC001 suggested an acceptable safety profile and supported evaluation of this agent in a FIH clinical trial.

[0452]

[0230] ADC001 Relevant Nonclinical Pharmacology, Pharmacokinetics, and Toxicology

[0231] ADC001 is an ADC comprised of a CEACAM5-targeting human IgGl conjugated to the TOPO-I inhibitor exatecan (DAR8) via a plasma-stable valine-citrulline cleavable linker. ADC001 selectively binds the membrane-proximal B3 domain of CEACAM5 and efficiently delivers exatecan intracellularly via lysosomal-mediated liberation by cathepsin-B. ADC001 induced DNA-damage. demonstrated by the induction of the DNA-damage-response pathway pharmacodynamic biomarkers pKAPl, pCHKl and H2Ax 24-hours post-treatment in vitro, with subsequent antigen-dependent cellular growth inhibition across cell lines expressing CEACAM5 from 25,000 to 150,000 receptors per cell. In heterogenous tumor settings. ADC001 also induced cellular growth inhibition on antigen negative cells within close proximity to CEACAM5+ cells via the secondary’ mechanism of bystander kill effect.

[0453]

[0232] FIGs. 1A-1D are a set of graphs showing in vivo efficacy of ADC001 in CRC CDX models (NCI-H508 FIG. IB; CL-40 FIG. 1C; and Lsl80 FIG. ID). (FIG. 1A) H-score for CEACAM5 in tumors from NCI-H508, CL-40, and Lsl80 compared to the CEACAM5 H-score in a human CRC TMA, and (FIGs. 1B-D) after a single bolus IV injection of BMS-986490 at 1, 3, or 10 mg / kg body weight (FIG. IB) or at 0.3, 1, 3, or 10 mg / kg body weight (FIG. 1C and FIG.1D) are shown. ADC001 demonstrated potent anti-tumor activity following single administration of 1.0, 3.0 or 10.0 mg / kg doses in multiple cell line derived xenograft gastric (MKN45), pancreatic (BxPC3), and CRC (NCI-H508, CL-40, Lsl80) tumor models and 2 patient-derived xenograft CRC tumor models (CTG-0899, CTG-0069). In a CRC PDX mouse clinical trial, 3.0 and 10.0 mg / kg doses of ADC001 resulted in disease control rates of 76% and 94%, respectively. Efficacy of ADCOOlwas observed across a broad range of CEACAM5 expression levels and with both MSS and MSLhigh genetic phenotypes. CEACAM5 expression in these CDX and PDX tumor models are representative of the spectrum of CEACAM5 expression in CRC patient population. Furthermore, in the MKN45 CDX model, tumor exposure of both ADC001 and the released payload, exatecan, increased within the first 72 hours and was sustained up to 240 hours post treatment.

[0454] Pharmacodynamic biomarkers for induction of DNA damage-response pathways

[0455]

[0456] Docket No. 14887-WO

[0457] downstream of TOPO-I inhibition were sustained for 336 hours post dose. Additionally, in the CRC CDX model NCI-H508 and CRC PDX model CTG-0835, ADCOOlin combination with anti-VEGF bevacizumab demonstrated superior efficacy to either agent as monotherapy. PK analysis of ADC001 supports a 3.0 mg / kg dose and Q3W dosing frequency in human patients.

[0458]

[0233] Mice with NCI-H508 tumors (n =7 per group) (A), Lsl80 tumors (n = 7 per group) (B), or CTG-0835 tumors (n = 5 per group) (C) were treated with a single bolus IV injection of ADC001 at 3 mg / kg, or twice weekly IP injection of bevacizumab at 20 mg / kg for 3 weeks, or combination of ADCOOland bevacizumab. Tumor volume was measured over time after tumor implant was indicated. Results are presented as mean values with error bars representing SEM. Statistical analysis was performed using student t test. **P < 0.01, ***P < 0.001. Data in FIG. 2A, FIG. 2B and FIG. 2C show the antitumor efficacy of ADC001 as a monotherapy and also in combination with bevacizumab in CRC CDX tumor models NCI-H508 and Lsl80, and PDX Model CTG- 0835.

[0459]

[0234] ADC001 linker stability7and payload release were evaluated in serum from C57BL / 6 mouse, cynomolgus monkey, and human, in plasma from rat and human, and in liver lysosomes from rat and human. ADC001 was stable in serum with minimal payload release (< 0.1% after 7-day incubation at 37°C). In liver lysosomal (rat and human) incubations with ADC001, exatecan was the only product formed in these incubations. These data clearly show that the ADC was stable in serum and the payload was released in the lysosome.

[0460]

[0235] Following a single IV dose of ADC001 (1 , 3, and 10 mg / kg) to the MKN45 or BxPC3 xenografted mice, ADC 001 demonstrated a dose-dependent exposure increase with low CLT, low Vss, and a long T-HALF. In a separate tissue-distribution study in MKN45 xenografted mice (3 mg / kg and 10 mg / kg), the tissue to plasma ratio of tADC was 0.11 for liver and 0.40 to 0.48 for tumor. The exposure of released payload (exatecan) in tumor was high with Cmax of 53 nM (23 ng / mL) and AUC(0-360h) of 10.3 pM»h (4.5 pg*h / mL) at 3 mg / kg and Cmax of 204 nM (89 ng / mL) and AUC(0-240h) of 39 pM*h (17 pg»h / mL) at 10 mg / kg. respectively. However, exatecan was not detected in plasma or liver. These results indicate target-mediated tumor distribution and drug disposition.

[0461]

[0236] Pharmacokinetics of ADC001 in cynomolgus monkeys exhibited PK characteristics ty pical of ADC in monkey7. Following IV administration of ADC001 to monkeys at 3.0,

[0462]

[0463] Docket No. 14887-WO

[0464] 10.0, and 30.0 mg / kg every 3 weeks (Q3W), the systemic exposures to total ADC (i.e., ADC001) and total antibody increased with increasing doses in a dose-proportional manner. Elimination of total ADC was slow with low clearance (0.007 ml / min / kg) and long T-HALF (4 - 6 days). The formation of anti-drug antibody (ADA) against ADC001 was detected in some animals (1 / 3 monkey at 3.0 mg / kg and 2 / 3 monkeys at 10.0 mg / kg from Day 8 to Day 43) and had minimal impact on systemic exposures.

[0465]

[0237] The human PK of ADC001 was predicted based on the PK profdes observed in cynomolgus monkeys. The predicted total plasma (or serum) clearance and Vss in humans for ADC001 were 0.0065 ml / min / kg and 3.65 L (0.052 L / kg), respectively. The predicted T-HALF was 4.4 days.

[0466]

[0238] Exatecan permeability, distribution, metabolism, and excretion were evaluated in a series of in vitro and in vivo preclinical studies. Based on in vitro data, exatecan is a highly permeable compound. The plasma protein binding is high in human (97%), similar to monkey (96%), higher than dog (86%) and rat (93%). Exatecan is metabolized mainly by cytochrome P450 (CYP) 3A4 / 5 and to a lesser extent by CYP1A2 leading to formation of Ml and M2, respectively. In plasma, Ml accounted for approximately 4% of total exposure. In urine, Ml / parent ratio was at approximately 2:1. Exatecan is a substrate of the efflux transporter P-gp. The principal route of excretion (IV dosing) in preclinical species was the fecal excretion (70 to 90%), whereas 10 to 25% of the dose was recovered in the urine. The results of the previously discussed animal studies with ADC001 showed that this ADC treatment had a strong anti-tumor efficacy as a monotherapy or a combination therapy with bevacizumab. In addition, data showed that the ADC treatment had an acceptable safety profile and supported evaluation of this agent in a FIH clinical trial in solid tumors.

[0467]

[0239] Example 3. A Phase l / 2a Study of ACD001 with or without bevacizumab in human participants with advanced solid tumors

[0468]

[0240] ADC001 is a humanized, IgGl ADC targeting CEACAM5 for tumor-directed delivery' of exatecan, a TOPO-I inhibitor. This Example describes a Phase l / 2a, FIH study of ADC001 in participants with select advanced solid tumors known or likely to express CEACAM5.

[0469]

[0241] Overall Design

[0470]

[0471] Docket No. 14887-WO

[0472]

[0242] Investigators are performing a Phase l / 2a, multi-center, open-label, first in human study of ADC001 as a monotherapy and in combination with bevacizumab in participants with select advanced solid tumors known to express CEACAM5. Initially, the study will conduct dose escalation and expansion of ADCOOlas monotherapy (Part 1A and Part 2 A) and in combination with bevacizumab (Part IB and Part 2B) in participants with CRC. Based on retrospective analysis of CEACAM5 expression by IHC and review of the totality of clinical data generated in CRC participants (Part 1 A and Part 2A). the Sponsor may open enrollment to other CE AC AM5 -expressing tumor types including NSCLC (Part 2A-NSCLC) and gastric cancer (Part 2A-GC). FIG. 3 is a study design schema for this study.

[0473]

[0243] Objectives and Endpoints

[0474]

[0244] The primary objective of Part 1 A, IB of the study is to determine the safety and tolerability of ADC001 up to MAD or MTD alone and in combination with bevacizumab in participants with advanced solid tumors to determine RDE and schedule of ADC01 alone and in combination with bevacizumab in participants with advanced solid tumors. Endpoints include analysis of the incidence of AEs, SAEs, AEs meeting protocol-defined DLT criteria, and AEs leading to discontinuation and deaths.

[0475]

[0245] The primary objectives of Part 1 A, IB, 2 A, 2B include defining the MTD, MAD and / or RDE. Endpoints include incidence of AEs, SAEs, AEs meeting protocol-defined DLT criteria, and AEs leading to discontinuation and deaths.

[0476]

[0246] Secondary objectives of Part 1A, IB include to characterize the PK profile of ADC001. total antibody, and exatecan following IV administration alone and in combination with bevacizumab in participants with advanced solid tumors. Endpoints include summary measures of PK parameters, such as, but not limited to, AUC(tau), Ctrough, Cmax, and Tmax, of ADC001, total antibody, and exatecan in monotherapy and in combination with bevacizumab.

[0477]

[0247] Secondary objectives of Part 2A, and Part 2B include evaluating the preliminary efficacy of ADC001 alone and in combination with bevacizumab in participants with advanced solid tumors. Endpoints include ORR, per RECIST vl.l by investigator.

[0478]

[0248] Number of Participants

[0479]

[0249] Part 1 A (ADC001 Monotherapy Dose Escalation): the maximum sample size is up to approximately 60 participants. Part IB (ADC001 Dose Escalation in Combination with Bevacizumab): the maximum sample size is up to approximately 60 participants.

[0480]

[0481] Docket No. 14887-WO

[0482]

[0250] Part 2A (ADC001 Monotherapy Dose Expansion in Participants with mCRC):

[0483] participants will be assigned to one of 2 treatment arms, each arm consisting of approximately 20 response evaluable participants (maximum up to 40). If additional participants are enrolled, then up to 80 participants will be treated in Part 2A.

[0484]

[0251] Part 2A-NSCLC / GC (ADC001 Monotherapy Dose Expansion in Participants with NSCLC and Gastric Cancer): at least one dose regimen will be evaluated in approximately 20 response evaluable participants (maximum up to 40) each with NSCLC (Part 2A-NSCLC) and gastric cancer (Part 2A-GC) to explore clinical activity, and additionally \ to further evaluate safety, tolerability, efficacy, pharmacodynamics, or subgroups of interest. If additional participants are enrolled (to a maximum of approximately 40 evaluable participants per tumor type), then up to 80 participants will be treated in Part 2A-NSCLC / GC.

[0485]

[0252] Part 2B (ADC001 Dose Expansion in Combination with Bevacizumab): Participants with CRC will be assigned to one of 2 treatment arms, each arm consisting of approximately 20 (maximum up to 40) response evaluable participants. If additional participants are enrolled, then up to 80 participants will be treated in Part 2B.

[0486]

[0253] The estimated number of evaluable participants with CRC across applicable study parts (Parts 1 A, IB, 2A, and 2B) will be up to a maximum of 280 and additional 80 participants with NSCLC and GC (in 2A-NSCLC and 2A-GC).

[0487]

[0254] Study Population

[0488]

[0255] Key Inclusion Criteria:

[0489] • Participants with a histological or cytological confirmed diagnosis of an advanced solid tumor as defined below and includes.

[0490] • Parts 1A, IB, 2A and 2B: Metastatic, recurrent, or unresectable CRC with adenocarcinoma histology treated with at least 1 line (no more than 4 lines for Part 2A) of systemic cancer therapy in the metastatic or adjuvant setting.

[0491] • Part 2A-NSCLC (participants with NSCLC) NSCLC with non-squamous histology7, who have progressed on at least 2 prior lines of therapy, are ineligible for, or intolerant to approved therapy(ies) known to provide clinical benefit for the condition of the participant. Participants must have received and progressed on or after anti-PD-(L)l therapy, if available.

[0492] • Part 2A-GC: Participants with advanced, or metastatic gastric or GEJ adenocarcinoma (including signet cell histology) who have progressed on at least 1 prior line of therapy,

[0493]

[0494] Docket No. 14887-WO

[0495] are ineligible for, or intolerant of approved therapy(ies) known to provide clinical benefit for the condition of the participant. Participants must have at least 1 lesion that meets the definition of measurable disease by RECIST vl.l.

[0496] • ECOG performance status of 0 or 1.

[0497]

[0256] Key Exclusion Criteria:

[0498]

[0257] Key exclusion criteria include: previously received therapy targeting CEACAM5; previously received ADCs with TOPO-1 inhibitor or tubulin inhibitor payloads; participants with a known history of anaphylactic reactions to irinotecan and / or bevacizumab (if received previously), and Grade > 3 ILD / Pneumonitis.

[0499]

[0258] Study Intervention Administration and Duration

[0500]

[0259] ADC001 is being investigated in this Phase l / 2a open label study in participants with selected advanced solid tumors. ADC001 will be administered intravenously IV on Day 1 of each cycle for up to 35 cycles (alternatively day 1 and / or day 15 in Q2W or Q4W schedule for up to 26 cycles). Each cycle will be 21 days for Q3W schedule and / or 28 days for Q2W or Q4W schedules. Study will include treatment with ADC001 for up to 104 weeks (approximately 2 years) and survival follow-up for up to 104 eeks (approximately 24 months) after the last dose of study intervention. Participants may be enrolled in the study for up to approximately 4 years (inclusive of screening, treatment, and follow-up periods).

[0501]

[0260] Statistical Methods

[0502]

[0261] Parts 1 A and IB will be guided by the BOIN method, where > 3 participants are treated per dose cohort. The selected target DLT rate is 25%, the escalation boundary is 22.4%, and the de-escalation boundary is 32.2%. The primary endpoints are to assess safety based on incidence of AEs, SAEs, AEs leading to discontinuation, and deaths, to assess safety up to MAD / MTD to determine RDE. In addition, the selected tolerable dose levels of ADC001 monotherapy (Part 1A) and ADC001 in combination with bevacizumab (Part IB) will be determined using the DLT rate within the 21-day (or 28- day with alternate dose schedule) window7among DLT-evaluable participants in Parts 1A and IB. The incidence of DLTs along with overall assessment of safety, PK and PD data will be calculated and compared to the actions recommended by the BOIN design to guide the dose escalation / stay / de-escalation decision for the ADC001 monotherapy (Part 1 A) and for the ADC001 in combination with bevacizumab (Part IB).

[0503]

[0504] Docket No. 14887-WO

[0505]

[0262] In Part 2, a Bayesian continuous safety monitoring framework will be utilized to monitor toxicity and detect safety signals affecting conduct of the study.

[0506]

Claims

1. Docket No. 14887-WO2.CLAIMS1. A method of treating cancer in a subject in need thereof, wherein the method comprises4.administering a therapeutically effective dose of an antibody drug conjugate (ADC) having the structure:

6.

7. ADC001,8.or a pharmaceutically acceptable salt, a stereoisomer, or a solvate thereof, wherein AB is an anti-CEAC AM5 antibody or antigen binding portion thereof comprising (a) a heavy chain variable region (VH) comprising complementarity determining region (CDR) 1 , CDR2, and CDR3 regions comprising the amino acid sequences set forth in SEQ ID NOs: 1, 2, and 3, respectively, and a light chain variable region (VL) comprising CDR1 , CDR2, and CDR3 regions comprising the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10.

2. The method of claim 1, wherein the anti-CEAC AM5 antibody or antigen-binding portion thereof is administered at a dose ranging of the ADC from about 0.5 mg / kg body weight to about 8 mg / kg body weight of the ADC.

3. The method of claim 2 comprising administering the ADC at a dose of about 1 mg / kg body weight.

4. The method of claim 2 comprising administering the ADC at a dose of about 2 mg / kg body weight.

5. The method of claim 2 comprising administering the ADC at a dose of about 2.5 mg / kg or 2.8 mg / kg body weight.

6. The method of claim 2 comprising administering the ADC at a dose of about 3 mg / kg body weight.

7. The method of claim 2 comprising administering the ADC at a dose of about 3.5 mg / kg body weight.

16.

17. Docket No. 14887-WO8. The method of claim 2 comprising administering the ADC at a dose of about 4 mg / kg body weight.

9. The method of claim 2 comprising administering the ADC at a dose of about 4.5 mg / kg body weight.

10. The method of claim 2 comprising administering the ADC at a dose of about 5 mg / kg body weight.

11. The method of claim 2 comprising administering the ADC at a dose of about 6 mg / kg body weight.

12. The method of claim 2 comprising administering the ADC at a dose of about 7 mg / kg body weight.

13. The method of claim 2 comprising administering the ADC at a dose of about 8 mg / kg body weight.

14. The method of any of claims 1-13, wherein the method comprises administering the ADC every two weeks (Q2W), every7three weeks (Q3W), or ADC every four weeks (Q4W).

15. The method of claim 14, wherein the method comprises administering the ADC Q3W.

16. The method of any one of claims 1-15, wherein the method comprises administering the ADC intravenously (IV).

17. The method of any one of claims 1-16, wherein a dosing cycle of administering the ADC comprises about 14 to about 28 days.

18. The method of any of claims 1-17, wherein the anti-CEACAM5 antibody or antigen binding portion thereof comprises a heavy chain variable region (VH) which comprises the amino acid sequence set forth in SEQ ID NOs: 4.

19. The method of any of claims 1-17, wherein the anti-CEACAM5 antibody or antigen binding portion thereof comprises a VL which comprises the amino acid sequence set forth in SEQ ID NO: 11.

20. The method of any of claims 1-17, wherein the anti-CEACAM5 antibody or antigen binding portion thereof comprises a VH and a VL which comprise the amino acid sequence set forth in SEQ ID NOs: 4 and the amino acid sequence set forth in SEQ ID NO: 11, respectively.

21. The method of claims 1-20, wherein the anti-CEACAM5 antibody or antigen binding portion thereof comprises an IgGl constant region, IgG2 constant region, IgG3 constant region, IgG4 constant region, or a variant thereof.

33.

34. Docket No. 14887-WO22. The method of claim 21, wherein the anti-CEACAM5 antibody comprises an IgGl antibody.

23. The method of claim 21 or 22, wherein the anti-CEACAM5 antibody or antigen binding portion thereof comprises an IgG1.3f constant region.

24. The ADC of any of claims 1-23, wherein the anti-CEACAM5 antibody is a human, humanized, or chimeric antibody.

25. The method of any of claims 1-24, wherein the anti-CEACAM5 antibody or antigen binding portion thereof comprising a heavy chain which comprises the amino acid sequence set forth in SEQ ID NOs: 6.

26. The method of any of claims 1 -24, wherein the anti-CEAC AM5 antibody or antigen binding portion thereof comprising a light chain which comprises the amino acid sequence set forth in SEQ ID NO: 13.

27. The method of any of claims 1-24. wherein the anti-CEAC AM5 antibody or antigen binding portion thereof comprises a heavy chain and a light chain which comprise the amino acid sequence set forth in SEQ ID NOs: 6 and the amino acid sequence set forth in SEQ ID NO: 13, respectively.

28. The method of any one of claims 1-27, wherein the antibody, or antigen binding portion thereof, comprises a bispecific molecule comprising the anti-CEACAM5 antibody or antigen binding portion thereof and a second binding region that binds to another antigen.

29. The method of any one of claims 1-28, wherein the antibody, or antigen binding portion thereof, comprises a multispecific molecule comprising the anti-CEACAM5 antibody, or antigen binding portion thereof and at least two binding regions, each of which binds other antigens.

30. The method of any of claims 1-29 comprising administering the ADC at a dose of 1 mg / kg body weight.

31. The method of any of claims 1-29 comprising administering the ADC at a dose of 2 mg / kg body weight.

32. The method of any of claims 1-29 comprising administering the ADC at a dose of 2.5 mg / kg body weight.

33. The method of any of claims 1-29 comprising administering the ADC at a dose of 3 mg / kg body weight.

34. The method of any of claims 1-29 comprising administering the ADC at a dose of 4 mg / kg body weight.

48.

49. Docket No. 14887-WO35. The method of any of claims 1-29 comprising administering the ADC at a dose of 5 mg / kg body weight.

36. The method of any of claims 1-29 comprising administering the ADC at a dose of 6 mg / kg body weight.

37. A method of treating cancer in a subject in need thereof, wherein the method comprises administering a therapeutically effective dose of an anti-VEGF treatment and an antibody drug conjugate (ADC) treatment; wherein53.(i) the anti-VEGF treatment comprises administering about 5 mg / kg body weight to about 7.5 mg / kg body weight of bevacizumab to the subject on about day 1 of a dosing cycle; and54.(ii) the ADC treatment comprises administering an ADC having the structure:

56.

57. ADC001,58.or a pharmaceutically acceptable salt, a stereoisomer, or a solvate thereof, wherein AB is an anti-CEACAM5 antibody or antigen binding portion thereof comprising (a) a heavy chain variable region (VH) comprising complementarity determining region (CDR)l, CDR2, and CDR3 regions comprising the amino acid sequences set forth in SEQ ID NOs: 1, 2, and 3, respectively, and a light chain variable region (VL) comprising CDR1, CDR2, and CDR3 regions comprising the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10.

38. The method of claim 37, wherein the anti-CEACAM5 antibody or antigen-binding portion thereof is administered at a dose ranging of the ADC from about 0.5 mg / kg body weight to about 8 mg / kg body weight of the ADC.

39. The method of claim 38 comprising administering about 1 mg / kg body weight of the ADC40. The method of claim 38 comprising administering about 2 mg / kg body weight of the ADC.

63.

64. Docket No. 14887-WO41. The method of claim 38 comprising administering about 2.5 mg / kg or 2.8 mg / kg body weight of the ADC.

42. The method of claim 38 comprising administering about 3 mg / kg body weight of the ADC.

43. The method of claim 38 comprising administering the ADC at a dose of about 3.5 mg / kg body weight.

44. The method of claim 38 comprising administering about 4 mg / kg body weight of the ADC.

45. The method of claim 38 comprising administering the ADC at a dose of about 4.5 mg / kg body weight.

46. The method of claim 38 comprising administering about 5 mg / kg body weight of the ADC treatment.

47. The method of claim 38 comprising administering about 6 mg / kg body weight of the ADC treatment.

48. The method of any of claims 37-47, wherein the method comprises administering the ADC every two weeks (Q2W), every three weeks (Q3W), or ADC every' four weeks (Q4W).

49. The method of claim 48, wherein the method comprises administering the ADC Q3W.

50. The method of any one of claims 37-49, wherein the method comprises administering the ADC intravenously (IV).

51. The method of any one of claims 37-50, wherein a dosing cycle of the ADC treatment comprises about 14 days to about 28 days.

52. The method of any of claims 37-51, wherein the anti-CEACAM5 antibody or antigen binding portion thereof comprises a VH which comprises the amino acid sequence set forth in SEQ ID NOs: 4.

53. The method of any of claims 37-51, wherein the anti-CEACAM5 antibody or antigen binding portion thereof comprises a VL which comprises the amino acid sequence set forth in SEQ ID NO: 11.

54. The method of any of claims 37-51, wherein the anti-CEACAM5 antibody or antigen binding portion thereof comprising a VH and a VL which comprise the amino acid sequence set forth in SEQ ID NOs: 4 and the amino acid sequence set forth in SEQ ID NO: 11, respectively.

80.

81. Docket No. 14887-WO55. The method of claims 37-54, wherein the anti-CEACAM5 antibody or antigen binding portion thereof comprises an IgGl constant region, IgG2 constant region, IgG3 constant region, IgG4 constant region, or a variant thereof.

56. The method of claim 55, comprising an IgGl antibody.

57. The method of claim 55 or claim 56, comprising an IgG1.3f constant region.

58. The method of any of claims 37-57, wherein the anti-CEACAM5 antibody is a human, humanized, or chimeric antibody .

59. The method of any of claims 37-58, anti-CEACAM5 antibody or antigen binding portion thereof comprising a heavy chain which comprises the amino acid sequence set forth in SEQ ID NOs: 6.

60. The method of any of claims 37-58, anti-CEACAM5 antibody or antigen binding portion thereof comprising a light chain which comprises the amino acid sequence set forth in SEQ ID NO: 13.

61. The method of any of claims 37-58, anti-CEACAM5 antibody or antigen binding portion thereof comprising a heavy chain and a light chain which comprise the amino acid sequence set forth in SEQ ID NOs: 6 and the amino acid sequence set forth in SEQ ID NO: 13, respectively.

62. The method of any one of claims 37-61, wherein the antibody, or antigen binding portion thereof, comprises a bispecific molecule comprising the anti-CEACAM5 antibody or antigen binding portion thereof and a second binding region that binds to another antigen.

63. The method of any one of claims 37-61, wherein the antibody, or antigen binding portion thereof, comprises a multispecific molecule comprising the anti-CEACAM5 antibody, or antigen binding portion thereof and at least two binding regions, each of which binds other antigens.

64. The method of any of claims 37-63 comprising administering the ADC at a dose of 1 mg / kg body weight.

65. The method of any of claims 37-63 comprising administering the ADC at a dose of 2 mg / kg body weight.

66. The method of any of claims 37-63 comprising administering the ADC at a dose of 2.5 mg / kg or 2.8 mg / kg body weight.

67. The method of any of claims 37-63 comprising administering the ADC at a dose of 3 mg / kg body weight.

96.

97. Docket No. 14887-WO68. The method of any of claims 37-63 comprising administering the ADC at a dose of about 3 mg / kg body weight.

69. The method of any of claims 37-63 comprising administering the ADC at a dose of 4 mg / kg body weight.

70. The method of any of claims 37-63 comprising administering the ADC at a dose of about 4.5 mg / kg body weight.

71. The method of any of claims 37-63 comprising administering the ADC at a dose of 5 mg / kg body weight.

72. The method of any of claims 37-63 comprising administering the ADC at a dose of 6 mg / kg body weight.

73. The method of any of claims 37-63 comprising administering the ADC at a dose of 7 mg / kg body weight.

74. The method of any of claims 37-63 comprising administering the ADC at a dose of 8 mg / kg body weight.

75. The method of any of claims 37-74, wherein the bevacizumab is administered intravenously.

76. The method of any of claims 37-75, wherein the bevacizumab is administered at a dose of 5 mg / kg body weight.

77. The method of any of claims 37-75, wherein the bevacizumab is administered at a dose of 7.5 mg / kg body weight.

78. The method of any of claims 37-75, wherein the bevacizumab is administered every two weeks (Q2W), or every three weeks (Q3W).

79. The method of any of claims 37-75, wherein 7.5 mg / kg body weight of bevacizumab is administered Q3W.

80. The method of any of claims 37-75, wherein 5.0 mg / kg body weight of bevacizumab is administered Q2W.111.81 The method of claim 79, wherein the ADC treatment is administered Q3W.

82. The method of claim 80, w herein the ADC treatment is administered Q2W.

83. The method of claim 80, wherein the ADC treatment is administered Q4W.

84. The method of any of claims 37-83, wherein the ADC and bevacizumab are administered for as long as clinical benefit is observed or until unmanageable toxicity or disease progression occurs.

116.

117. Docket No. 14887-WO85. The method of any of claims 37-84, wherein the ADC and bevacizumab are administered for at least 104 weeks total.

86. The method of any claims 37-85, wherein the ADC and bevacizumab are formulated for intravenous administration.

87. The method of any of claims 37-86, wherein the ADC and bevacizumab are administered sequentially to the subject.

88. The method of any of claims 87, wherein the ADC and bevacizumab are administered sequentially to the subject during the induction phase.

89. The method of any of claims 37-88, wherein the ADC is administered before the bevacizumab is administered.

90. The method of any of claims 37-88, wherein the bevacizumab is administered before the ADC is administered.

91. The method of any of claims 37-88, wherein the ADC and the bevacizumab are administered concurrently in separate compositions.

92. The method of any of claims 37-88, wherein the ADC and the bevacizumab are admixed as a single composition for concurrent administration.

93. The method of any of claims 37-88, wherein the ADC and bevacizumab are administered within about 30 minutes to about 60 minutes of each other.

94. The method of any of claims 1-93, wherein the cancer is a solid tumor cancer.

95. The method of any one of claims 1-93, wherein the cancer is selected from the group consisting of non-small cell lung cancer (NSCLC). gastric cancer, and colorectal carcinoma (CRC).

96. The method of claim 95, wherein the cancer is CRC.

97. The method of claim 96, wherein the CRC is metastatic, recurrent, or unresectable CRC.

98. The method of claim 96 or 97, wherein the CRC comprises adenocarcinoma histology treated with at least one line of systemic cancer therapy in the metastatic or adjuvant setting.

99. The method of claim 95, wherein the cancer is NSCLC.

100. The method of claim 99, wherein the NSCLC is histologically confirmed NSCLC meeting stage criteria for Stage IIIB, Stage IV, or recurrent disease.

101. The method of claim 99 or 100, wherein the NSCLC has progressed on at least 2 prior lines of therapy.

102. A kit for treating a subject afflicted with a cancer, the kit comprising:

137.

138. Docket No. 14887-WO139.(a) a dosage range of about 05 mg / kg body weight to about 8 mg / kg body weight an antibody drug conjugate (ADC) having the structure:

141.

142. ADC001,143.or a pharmaceutically acceptable salt, a stereoisomer, or a solvate thereof, wherein144.AB is an anti-CEACAM5 antibody or antigen binding portion thereof comprising (a) a heavy chain variable region (VH) comprising complementarity determining region (CDR)l, CDR2, and CDR3 regions comprising the amino acid sequences set forth in SEQ ID NOs: 1, 2, and 3, respectively, and a light chain variable region (VL) comprising CDR1, CDR2, and CDR3 regions comprising the amino acid sequences set forth in SEQ ID NOs: 8, 9, and 10;145.(b) optionally, a dosage ranging from about 5 mg / kg body weight to about 7.5 mg / kg body weight of bevacizumab; and146.(c) instructions for treating cancer as described in any of claims 1-101.

103. The kit of claim 102, wherein the anti-CEACAM5 antibody or antigen binding portion thereof comprises a VH which comprises the amino acid sequence set forth in SEQ ID NOs: 4.

104. The kit of claim 102, wherein the anti-CEACAM5 antibody or antigen binding portion thereof comprises a VL which comprises the amino acid sequence set forth in SEQ ID NO: 11.

105. The kit of claim 102. wherein the anti-CEACAM5 antibody or antigen binding portion thereof comprises a VH and a VL which comprise the amino acid sequence set forth in SEQ ID NOs: 4 and the amino acid sequence set forth in SEQ ID NO: 11, respectively.

106. The kit of any of claims 102-105, wherein the anti-CEACAM5 antibody or antigen binding portion thereof comprising a heavy chain which comprises the amino acid sequence set forth in SEQ ID NOs: 6.

152.

153. Docket No. 14887-WO107. The kit of any of claims 102-105, wherein the anti-CEACAM5 antibody or antigen binding portion thereof comprising a light chain which comprises the amino acid sequence set forth in SEQ ID NO: 13.

108. The kit of any of claims 102-105, wherein the anti-CEACAM5 antibody or antigen binding portion thereof comprising a heavy chain and a light chain which comprise the amino acid sequence set forth in SEQ ID NOs: 6 and the amino acid sequence set forth in SEQ ID NO: 13, respectively.

109. The kit of any of claims 102-108, wherein the ADC is formulated for intravenous administration.158.

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