WO2017160568A1

WO2017160568A1 - Combination therapy comprising the cdk4/6 inhibitor necitumumab and the egfr inhibitor abemaciclib for use in treating cancer

Info

Publication number: WO2017160568A1
Application number: PCT/US2017/021475
Authority: WO
Inventors: Richard P. BECKMANN; Maciej W. GIL; Ruslan D. NOVOSIADLY
Original assignee: Eli Lilly And Company
Priority date: 2016-03-16
Filing date: 2017-03-09
Publication date: 2017-09-21

Abstract

The present invention provides medicaments for use in treating cancer, in particular non-small cell lung cancer, colorectal cancer, head and neck cancer, or pancreatic carcinoma, comprising a CDK inhibitor, preferably abemaciclib, or a pharmaceutically acceptable salt thereof, in combination with an anti-EGFR antibody, preferably necitumumab.

Description

COMBINATION THERAPY COMPRISING THE CDK4/6 INHIBITOR NECITUMUMAB AND THE EGFR INHIBITOR ABEMACICLIB FOR USE IN TREATING CANCER

The present invention relates to the field of medicine. More preferably, the present invention relates to combinations of an inhibitor of cyclin dependent kinases 4 and 6 (CDK4/6) with an antibody against human epidermal growth factor receptor (EGFR), preferably, combinations of abemaciclib with necitumumab, and to methods of using the combinations to treat cancer, particularly lung cancer, more particularly, non-small cell lung cancer (NSCLC), as well as colorectal cancer, head and neck cancer, and pancreatic carcinoma.

Overexpression of EGFR has been reported in numerous human malignant conditions, including cancers of the bladder, brain, head and neck, pancreas, lung, breast, ovary, colon, prostate, and kidney. In many of these conditions, the overexpression of EGFR correlates or is associated with poor prognosis of the patients.

Lung cancer ranks as one of the most common causes of death due to cancer in both men and women throughout the world. The two main types of lung cancer are small cell lung cancer and NSCLC. NSCLC makes up approximately 85% or more of lung cancer cases. Treatment can involve surgery, chemotherapy, radiation therapy, biologic therapy, targeted therapy, immunotherapy, as well as selected combinations of these treatments. Unfortunately, a cure for NSCLC still remains elusive and there exists a need for more and different therapies that may prove to be effective in treating NSCLC.

Abemaciclib (LY2835219), [5-(4-ethyl-piperazin- l-ylmethyl)-pyridin-2-yl]-[5- fluoro-4-(7-fluoro-3-isopropyl-2-methyl-3H-benzoimidazol-5-yl)-pyrimidin-2-yl]-amine, is a CDK inhibitor that targets the cell cycle by inhibiting CDK4 and CDK6, with antineoplastic activities. Abemaciclib, including salt forms, and methods of making and using this compound including for the treatment of cancer and more preferably for the treatment of

NSCLC are disclosed in WO2010/075074. Furthermore, clinical investigations are ongoing for the compound as a single agent or in various combinations in patients with a variety of indications including NSCLC. The present invention provides for abemaciclib in various aspects disclosed herein. Necitumumab (IMC-11F8) is a recombinant human IgGl monoclonal antibody that binds to the extracellular domain III of the human EGFR and blocks interaction between EGFR and its ligands. Necitumumab (including its variable regions) and methods of making and using this antibody including for the treatment of neoplastic diseases such as solid and non-solid tumors are disclosed in WO2005/090407. Furthermore, necitumumab has been approved by the U.S. FDA and European Medical Authority for treating metastatic squamous NSCLC in combination with gemcitabine and cisplatin. The present invention provides for necitumumab in various aspects disclosed herein.

Novel methods of use of the combination of abemaciclib, or a pharmaceutically acceptable salt thereof, and necitumumab to treat a variety of cancers including squamous NSCLC, non-squamous NSCLC, colorectal cancer, head and neck cancer, and pancreatic carcinoma are presented herein. Broad combinations of a 'CDK inhibitor' with a compound capable of inhibiting ErbB, and in some cases EGFR, have been contemplated pre-clinically (e.£.,WO2012/123889 and WO2008/122779 which are both broadly directed to 'CDK inhibitors'). However, CDK inhibitors are a broad class of compounds and there is no teaching specific for using a CDK inhibitor selective for CDK4 and CDK6 with an EGFR inhibitor.

The present invention provides a combination therapy of abemaciclib, or a pharmaceutically acceptable salt thereof, and necitumumab for treating cancer, preferably squamous NSCLC, non-squamous NSCLC, colorectal cancer, head and neck cancer, and pancreatic carcinoma, more preferably non-squamous NSCLC and squamous NSCLC and most preferably non-squamous NSCLC, that provides enhanced and/or unexpected beneficial therapeutic effects from the combined activity of the agents. Additionally, the use of necitumumab in non-squamous NSCLC is particularly unexpected based on the results of the INSPIRE trial, which studied Portrazza® in non-squamous non-small cell lung cancer population, and the reference of these data in both the US and EP labels of Portrazza® ("Limitation of Use" and "Warnings and Precautions" section respectively).

Accordingly, in some aspects, the present invention provides a method of treating cancer in a patient, wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma, comprising administering to the patient in need of such a treatment an effective amount of necitumumab in combination with an effective about of abemaciclib, or a pharmaceutically acceptable salt thereof.

The present invention also provides for a method of treating cancer in a patient, wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma, comprising administering to the patient in need of such a treatment, a combination of necitumumab and abemaciclib, or a pharmaceutically acceptable salt thereof, wherein necitumumab is administered at a dose of 800 mg, and wherein abemaciclib is administered at a dose of 50 mg to 200 mg. More particularly, necitumumab is administered at a dose of 800 mg on days 1 and 8 of each 3- week cycle by intravenous infusion. More particularly, abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered twice a day. Yet more particularly, abemaciclib, or a pharmaceutically acceptable salt thereof, is administered at a dose of 50 mg. Yet more particularly, abemaciclib, or a pharmaceutically acceptable salt thereof, is administered at a dose of 100 mg. Yet more particularly, abemaciclib, or a pharmaceutically acceptable salt thereof, is administered at a dose of 150 mg. Yet more particularly, abemaciclib, or a pharmaceutically acceptable salt thereof, is administered at a dose of 200 mg. Even more particularly, the combination of necitumumab and abemaciclib, or a pharmaceutically acceptable salt thereof, is administered simultaneously, separately, or sequentially.

The present invention also provides for necitumumab for use in simultaneous, separate, or sequential combination with abemaciclib, or a pharmaceutically acceptable salt thereof, in the treatment of cancer, wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma. More particularly, necitumumab is administered at a dose of 800 mg by intravenous infusion. More particularly, abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered at a dose of 50 mg to 200 mg. More particularly,

necitumumab is administered at a dose of 800 mg by intravenous infusion and wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered at a dose of 50 mg to 200 mg. More particularly, necitumumab is administered at a dose of 800 mg on days 1 and 8 of each 3 -week cycle by intravenous infusion and wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered at a dose of 50 mg to 200 mg twice a day.

The present invention also provides for abemaciclib, or a pharmaceutically acceptable salt thereof, for use in simultaneous, separate, or sequential combination with necitumumab for the treatment of cancer, wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered at a dose of 50 mg to 200 mg and wherein necitumumab is administered at a dose of 800 mg by intravenous infusion, and wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma. More particularly, abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered at a dose of 50 mg to 200 mg twice a day and wherein necitumumab is administered at a dose of 800 mg by intravenous infusion on days 1 and 8 of each 3-week cycle. Yet more particularly, abemaciclib, or a pharmaceutically acceptable salt thereof, is administered at a dose of 50 mg. Yet more particularly, abemaciclib, or a pharmaceutically acceptable salt thereof, is administered at a dose of 100 mg. Yet more particularly, abemaciclib, or a pharmaceutically acceptable salt thereof, is administered at a dose of 150 mg. Yet more particularly, abemaciclib, or a pharmaceutically acceptable salt thereof, is administered at a dose of 200 mg.

The present invention also provides for use of necitumumab in the manufacture of a medicament for the treatment of cancer, wherein necitumumab is administered at a dose of 800 mg by intravenous infusion in simultaneous, separate, or sequential combination with abemaciclib, or a pharmaceutically acceptable salt thereof, wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered at a dose of 50 mg to 200 mg and wherein the cancer is selected from the group consisting of non- small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma.

The present invention also provides for use of necitumumab in the manufacture of a medicament for the treatment of cancer, wherein necitumumab is administered at a dose of 800 mg on days 1 and 8 of each 3-week cycle by intravenous infusion in separate, or sequential combination with abemaciclib, or a pharmaceutically acceptable salt thereof, wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered at a dose of 50 mg to 200 mg twice a day, and wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma.

The present invention also provides for use of abemaciclib in the manufacture of a medicament for the treatment of cancer, wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered at a dose of 50 mg to 200 mg in simultaneous, separate, or sequential combination with necitumumab, wherein necitumumab is

administered at a dose of 800 mg by intravenous infusion, and wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma.

The present invention further provides for use of abemaciclib in the manufacture of a medicament for the treatment of cancer, wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered at a dose of 50 mg to 200 mg twice a day in separate, or sequential combination with necitumumab, wherein necitumumab is

administered at a dose of 800 mg by intravenous infusion on days 1 and 8 of each 3-week cycle by intravenous infusion, and wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma.

The present invention provides for a kit comprising necitumumab and abemaciclib, or a pharmaceutically acceptable salt thereof, for the treatment of cancer, wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma. More particularly, the amount of necitumumab is 800 mg, and the amount of abemaciclib is 50 mg to 200 mg. Yet more particularly, the amount of abemaciclib, or a pharmaceutically acceptable salt thereof, is 50 mg. Yet more particularly, the amount of abemaciclib, or a pharmaceutically acceptable salt thereof, is 100 mg. Yet more particularly, the amount of abemaciclib, or a pharmaceutically acceptable salt thereof, is 150 mg. Yet more particularly, the amount of abemaciclib, or a pharmaceutically acceptable salt thereof, is 200 mg.

The present invention also provides a kit, comprising a pharmaceutical composition, comprising necitumumab with one or more pharmaceutically acceptable carriers, diluents, or excipients, and a pharmaceutical composition, comprising abemaciclib, or a pharmaceutically acceptable salt thereof, with one or more pharmaceutically acceptable carriers, diluents, or excipients for the treatment of cancer, and wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma. More particularly, the amount of necitumumab is 800 mg, and wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is 50 mg to 200 mg. Yet more particularly, the amount of abemaciclib, or a pharmaceutically acceptable salt thereof, is 50 mg. Yet more particularly, the amount of abemaciclib, or a pharmaceutically acceptable salt thereof, is 100 mg. Yet more particularly, the amount of abemaciclib, or a pharmaceutically acceptable salt thereof, is 150 mg. Yet more particularly, the amount of abemaciclib, or a pharmaceutically acceptable salt thereof, is 200 mg.

The present invention also provides a kit comprising necitumumab and abemaciclib, or a pharmaceutically acceptable salt thereof, wherein necitumumab is formulated at a final concentration of 16 mg/mL in a formulation of 10 mM citrate, 40 mM sodium chloride, 133 mM glycine, 50 mM mannitol, 0.01% polysorbate-80, pH6.0; and wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is supplied as capsules or tablets of 50 mg of abemaciclib per capsule or tablet.

The present invention also provides a combination of necitumumab and abemaciclib, or a pharmaceutically acceptable salt thereof, for simultaneous, separate, or sequential use in the treatment of cancer, wherein the cancer is selected from the group consisting of non- small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma.

More particularly, necitumumab is administered at a dose of 800 mg by intravenous infusion. More particularly, abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered at a dose of 50 mg to 200 mg. Yet more particularly, abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered at a dose of 50 mg. Yet more particularly, abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered at a dose of 100 mg. Yet more particularly, abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered at a dose of 150 mg. Yet more particularly, abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered at a dose of 200 mg.

In a preferred aspect of the invention relating to the various methods disclosed above, necitumumab for the use of disclosed above, abemaciclib for the use of disclosed above, the various kits disclosed above, and/or the various combinations disclosed above, the non-small cell lung cancer is squamous.

In a preferred aspect of the invention relating to the various methods disclosed above, necitumumab for the use of disclosed above, abemaciclib for the use of disclosed above, the various kits disclosed above, and/or the various combinations disclosed above, the non-small cell lung cancer is non-squamous.

In a preferred aspect of the invention relating to the various methods disclosed above, necitumumab for the use of disclosed above, abemaciclib for the use of disclosed above, the various kits disclosed above, and/or the various combinations disclosed above, the head and neck cancer is squamous.

One aspect of the present invention is treating cancer. In a preferred aspect, the cancer is NSCLC, colorectal cancer, head and neck cancer, or pancreatic carcinoma. In a more preferred aspect, the NSCLC is non-squamous or squamous. In a most preferable aspect the NSCLC is non-squamous. In a preferred aspect the head and neck cancer is squamous.

The present invention also contemplates the following non-limiting list of aspects, which are further described elsewhere herein:

In some aspects, the compounds and/or medicaments are administered

simultaneously, separately, or sequentially with one or more anti-tumor agents selected from the group consisting of cisplatin, carboplatin, dacarbazine, liposomal doxorubicin, docetaxel, cyclophosphamide, doxorubicin, gemcitabine, navelbine, eribulin, paclitaxel, paclitaxel protein-bound particles for injectable suspension, ixabepilone, capecitabine, and FOLFOX (leucovorin, fluorouracil, and oxaliplatin), FOLFIRI (leucovorin, fluorouracil, and irinotecan).

In some aspects, the cancer is NSCLC, colorectal cancer, head and neck cancer, or pancreatic carcinoma, stomach, skin and ovary.

In a preferred aspect, the present invention also provides for a method of treating cancer in a patient, wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma, comprising administering to the patient in need of such a treatment, a combination of necitumumab and abemaciclib, or a pharmaceutically acceptable salt thereof, wherein necitumumab is administered at a dose of 800 mg, and wherein abemaciclib is administered at a dose of 150 mg.

In a preferred aspect, the present invention also provides for a method of treating cancer in a patient, wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma, comprising administering to the patient in need of such a treatment, a combination of necitumumab and abemaciclib, or a pharmaceutically acceptable salt thereof, wherein necitumumab is administered at a dose of 800 mg on days 1 and 8 of each 3 -week cycle by intravenous infusion, and wherein abemaciclib is orally administered twice a day at a dose of 150 mg.

In a preferred aspect, the present invention also provides for necitumumab for use in simultaneous, separate, or sequential combination with abemaciclib, or a pharmaceutically acceptable salt thereof, in the treatment of cancer, wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma, wherein necitumumab is administered at a dose of 800 mg by intravenous infusion, and wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered at a dose of 150 mg.

In a preferred aspect, the present invention also provides for necitumumab for use in simultaneous, separate, or sequential combination with abemaciclib, or a pharmaceutically acceptable salt thereof, in the treatment of cancer, wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma wherein necitumumab is administered at a dose of 800 mg on days 1 and 8 of each 3-week cycle by intravenous infusion, and wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered twice a day at a dose of 150 mg.

In a preferred aspect, the present invention also provides for use of necitumumab in the manufacture of a medicament for the treatment of cancer, wherein necitumumab is administered at a dose of 800 mg by intravenous infusion in simultaneous, separate, or sequential combination with abemaciclib, or a pharmaceutically acceptable salt thereof, wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered at a dose of 150 mg and wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma.

In a preferred aspect, the present invention also provides for use of necitumumab in the manufacture of a medicament for the treatment of cancer, wherein necitumumab is administered at a dose of 800 mg on days 1 and 8 of each 3 -week cycle by intravenous infusion in separate, or sequential combination with abemaciclib, or a pharmaceutically acceptable salt thereof, wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered at a dose of 150 mg twice a day, and wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma.

In a preferred aspect, the present invention also provides for use of abemaciclib in the manufacture of a medicament for the treatment of cancer, wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered at a dose of 150 mg in simultaneous, separate, or sequential combination with necitumumab, wherein necitumumab is administered at a dose of 800 mg by intravenous infusion, and wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma.

In a preferred aspect, the present invention also provides for use of abemaciclib in the manufacture of a medicament for the treatment of cancer, wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered at a dose of 150 mg twice a day in separate, or sequential combination with necitumumab, wherein necitumumab is administered at a dose of 800 mg by intravenous infusion on days 1 and 8 of each 3-week cycle by intravenous infusion, and wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma.

In a preferred aspect, the present invention also provides for a kit comprising necitumumab and abemaciclib, or a pharmaceutically acceptable salt thereof, for the treatment of cancer, wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma wherein the amount of necitumumab is 800 mg, and the amount of abemaciclib is 150 mg.

In a preferred aspect, the present invention also provides for a kit, comprising a pharmaceutical composition, comprising necitumumab with one or more pharmaceutically acceptable carriers, diluents, or excipients, and a pharmaceutical composition, comprising abemaciclib, or a pharmaceutically acceptable salt thereof, with one or more

pharmaceutically acceptable carriers, diluents, or excipients for the treatment of cancer, and wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma wherein the amount of necitumumab is 800 mg, and wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is 150 mg to 200 mg.

In a preferred aspect, the present invention also provides for a combination of necitumumab and abemaciclib, or a pharmaceutically acceptable salt thereof, for simultaneous, separate, or sequential use in the treatment of cancer, wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma wherein necitumumab is administered at a dose of 800 mg by intravenous infusion and wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered at a dose of 150 mg.

In a preferred aspect, the present invention also provides for a combination of necitumumab and abemaciclib, or a pharmaceutically acceptable salt thereof, for simultaneous, separate, or sequential use in the treatment of cancer, wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma wherein necitumumab is administered at a dose of 800 mg on days 1 and 8 of each 3-week cycle by intravenous infusion and wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered at a dose of 150 mg twice a day.

"Cyclin-dependent kinases" or "CDKs" drive cell cycle progression and control transcriptional processes. The dysregulation of multiple CDK family members occurs commonly in human cancer; in particular, the cyclin D-CDK4/6-retinoblastoma protein (RB)-INK4 axis is universally disrupted, facilitating cancer cell proliferation and prompting interest in targeting CDK4/6 as an anticancer strategy. Most agents that have been tested inhibit multiple cell cycle and transcriptional CDKs and have carried toxicity. However, several selective and potent inhibitors of CDK4/6 have recently entered clinical trial.

Abemaciclib (LY2835219), [5-(4-ethyl-piperazin- l-ylmethyl)-pyridin-2-yl]-[5- fluoro-4-(7-fluoro-3-isopropyl-2-methyl-3H-benzoimidazol-5-yl)-pyrimidin-2-yl]-amine, is a CDK inhibitor that targets the cell cycle by inhibiting CDK4 and CDK6, with antineoplastic activities. The invention provides for abemaciclib in various aspects disclosed herein.

Abemaciclib is a CDK4 and CDK6 inhibitor having the following structure

and molecular formula of C27H32F2N8. Abemaciclib and salts thereof are disclosed

WO2010/075074. "Epidermal growth factor receptor" or "EGFR" is a member of the ErbB

(erythroblastic leukemia viral oncogene homolog) family of receptor tyrosine kinases. EGFR activation occurs in response to ligand stimulation and/or genetic alterations of the EGFR gene, such as somatic mutations, amplifications, or deletions. Activated EGFR induces downstream signaling through the MAPK (mitogen-activated protein kinases), PI3K/AKT (phosphoinositide 3-kinase/v-Akt murine thymoma viral oncogene), and PLCy

(phospholipase Cy) signal transduction pathways that mediate cell proliferation, cell survival, and cell migration, respectively, thereby contributing to neoplastic transformation and tumor growth.

Necitumumab is a recombinant IgGl human monoclonal antibody designed to bind and block the ligand binding site of EGFR. As used herein, the term "necitumumab" is also known as Portrazza®, IMC-11F8, and CAS registry number 906805-06-9. The invention provides for necitumumab in various aspects disclosed herein. Necitumumab is an antibody specific for human EGFR and comprises a heavy chain variable region (VH) having the amino acid sequence:

QVQLQES GPGLVKPS QTLS LTCTVS GGS IS S GD YYWS WIRQPPGKGLEWIG YIYYS GS TDYNPSLKSRVTMSVDTSKNQFSLKVNSVTAADTAVYYCARVSIFGVGTFDYWGQ GTLVTVSS (SEQ ID NO: l) and a light chain variable region (VL) having the amino acid sequence:

EIVMTQS P ATLS LS PGERATLS CR AS QS VS S YLA W YQQKPGQ APRLLIYD AS NRATGI P ARFS GS GS GTDFTLTIS S LEPEDF A V Y YCHQ YGS TPLTFGGGTKAEIK (SEQ ID NO:2). WO2005/090407.

As used herein, the terms "treating," "to treat," or "treatment" refers to restraining, slowing, stopping, reducing, or reversing the progression or severity of an existing symptom, disorder, condition, disease, or cancer.

As used herein, the term "patient" refers to a mammal, preferably a human.

As used herein, the terms "cancer" and "cancerous" refers to or describes the physiological condition in patients that is typically characterized by unregulated cell growth. Included in this definition are benign and malignant cancers. Examples of cancer include, but are not limited to, prostate cancer, breast cancer, triple negative breast cancer, colon cancer, lung cancer, NSCLC, head and neck cancer, melanoma, gastric cancer, pancreatic cancer, ovarian cancer, renal cell carcinoma, and hepatic cancer.

Provided herein are methods for treating NSCLC using necitumumab and

abemaciclib, or a pharmaceutically acceptable salt thereof. There are three main subtypes of NSCLC: squamous cell carcinoma, adenocarcinoma, and large cell (undifferentiated) carcinoma. Other subtypes include adenosquamous carcinoma and sarcomatoid carcinoma. Non-squamous NSCLC is a group of genetically diverse tumors that may include genetic aberrations such as KRAS, EGFR, ALK translocation, and the ROS 1 fusion. Targeted therapies are only currently approved for treatment of non-squamous NSCLC with EGFR mutations and ALK translocation genetic aberrations whereas no targeted agents have been approved for treatment of EGFR wild-type/ ALK translocation-negative non-squamous NSCLC.

As used herein, the term "kit" refers to a package comprising at least two separate containers wherein a first container contains abemaciclib, or a pharmaceutically acceptable salt thereof, and a second container contains necitumumab. A "kit" may also include instructions to administer all or a portion of the contents of these first and second containers to a cancer patient.

A potential advantage of the combination treatments of the invention is the possibility of producing marked and/or prolonged anti-cancer effects in a patient with an acceptable safety profile including acceptable tolerability, toxicities and/or adverse events , so that the patient benefits from the combination treatment method overall. The efficacy of the combination treatment of the invention can be measured by various endpoints commonly used in evaluating cancer treatments, including but not limited to, tumor regression, tumor weight or size shrinkage, time to progression, duration of survival, progression free survival, overall response rate, duration of response, and quality of life. The therapeutic agents used in the invention may cause inhibition of metastatic spread without shrinkage of the primary tumor, or may simply exert a tumoristatic effect. Because the invention relates to the use of a unique combination of anti-tumor agents, various approaches to determining efficacy of any particular combination therapy of the present invention can be optionally employed, including, for example, cell-cycle dependent biomarkers measurement/visualization, and measurement of response through radiological imaging.

As used herein, the term Complete Response (CR) is defined as the disappearance of all target lesions. Any pathological lymph nodes (whether target or non-target) must have reduction in short axis to <10 mm. Tumor marker results must have normalized.

As used herein, the term Partial Response (PR) is defined as at least a 30% decrease in the sum of diameter of target lesions, taking as reference the baseline sum diameters.

As used herein the term Progressive Disease (PD) is defined as at least a 20% increase in the sum of the diameters of target lesions, taking as reference the smallest sum on study (including the baseline sum if that is the smallest). In addition to the relative increase of 20%, the sum must also demonstrate an absolute increase of at least 5 mm. The appearance of 1 or more new lesions is also considered progression. For equivocal findings of progression (for example, very small and uncertain new lesions; cystic changes or necrosis in existing lesions), treatment may continue until the next scheduled assessment. If at the next scheduled assessment, progression is confirmed, the date of progression should be the earlier date when progression was suspected.

As used herein, the term Stable Disease (SD) is defined as neither sufficient shrinkage to qualify for PR nor sufficient increase to qualify for PD, taking as reference the smallest sum diameters while on study.

As used herein, the term Not Evaluable (NE) is define as when an incomplete radiologic assessment of target lesions is performed or there is a change in the method of measurement from baseline that impacts the ability to make a reliable evaluation of response. As used herein, the term Progression-Free Survival (PFS) is defined as the time from the date of enrollment until the date of radiographically documented PD or death due to any cause, whichever is earlier.

As used herein, the term Overall Survival (OS) is defined as the time from the date of study enrollment to the date of death from any cause.

As used herein, the term Objective Response Rate is defined as the proportion of patients achieving a best overall response of PR or CR.

As used herein, the term Disease Control Rate (DCR) is defined as the proportion of patients achieving a best overall response of SD, PR, or CR. As used herein, the term Overall Response Rate is based on each patient's best objective response will be determined for all patients evaluable via the Response Evaluation Criteria in Solid Tumors (RECIST) v 1.1 criteria. The Overall Response Rate (%) will be calculated as the number of patients with best objective response of CR or PR divided by the number of patients with measurable disease at baseline. The best objective response for a given patient will be based on objective responses determined from data obtained up to: progression, the last evaluable assessment in the absence of progression, or initiation of subsequent anticancer therapy. Patients for whom an objective response cannot be determined or for who the best objective response is NE will be considered non-responders. The Overall Response Rate will be summarized along with the 95% Clopper Pearson confidence interval.

As used herein, the term "effective amount" refers to the amount or dose of abemaciclib, or a pharmaceutically acceptable salt thereof, and/or to the amount or dose of necitumumab which, upon single or multiple dose administration to the patient, provides an effective response in the patient under diagnosis or treatment. It is also understood that a combination therapy of the present invention is carried out by administering abemaciclib, or a pharmaceutically acceptable salt thereof, together with necitumumab in any manner which provides effective levels of abemaciclib or a pharmaceutically acceptable salt thereof, and necitumumab in the body. An effective amount can be readily determined by the attending diagnostician, as one skilled in the art, by the use of known techniques and by observing results obtained under analogous circumstances. In determining the effective amount for a patient, a number of factors are considered by the attending diagnostician, including, but not limited to: the species of patient; its size, age, and general health; the specific disease or disorder involved; the degree of or involvement or the severity of the disease or disorder; the response of the individual patient; the particular compound administered; the mode of administration; the bioavailability characteristics of the preparation administered; the dose regimen selected; the use of concomitant medication; and other relevant circumstances.

As used herein, the term "effective response" of a patient or a patient's

"responsiveness" to treatment with a combination of agents and similar wording refers to the clinical or therapeutic benefit imparted to a patient upon co-administration of abemaciclib, or a pharmaceutically acceptable salt thereof, and necitumumab. Such benefit includes any one or more of: extending survival (including OS and PFS); resulting in an objective response (including a CR or a PR); or improving signs or symptoms of cancer, etc.

Abemaciclib, or a pharmaceutically acceptable salt thereof, is generally effective over a wide dosage range in the combination of the present invention. For example, dosages per day normally fall within the range of about 50 mg to about 400 mg twice per day or every 12 hours, preferably about 50 mg to about 200 mg twice per day, more preferably about 150 mg to about 200 mg twice per day, and most preferably about 150 mg. In addition, necitumumab is generally effective over a wide dosage range in the combination of the present invention. For example, dosages per 21-day cycle normally fall within the range of about 400 to 1000 mg with dosages on 2 or 3 days of that cycle, alternatively with dosages one time a week or once every two weeks, preferably about 400 to 1000 mg on Day 1, Day 8, and Day 15 of each 21 -day cycle, more preferably about 600 to 900 mg on Day 1 and Day 8 of each 21 -day cycle, and most preferably about 800 mg on Day 1 and Day 8 of each 21 -day cycle. In some instances dosage levels below the lower limit of the aforesaid ranges for abemaciclib, or a pharmaceutically acceptable salt thereof, and necitumumab, may be more than adequate, while in other cases smaller or still larger doses may be acceptably employed, and therefore the above dosage range is not intended to limit the scope of the invention in any way. When given in combination with necitumumab, for example, over a 21 -day cycle, abemaciclib, or a pharmaceutically acceptable salt thereof, is administered daily within the range of 50 mg to about 400 mg twice per day, preferably about 50 mg to about 200 mg twice per day, more preferably about 150 mg to about 200 mg twice per day, and most preferably about 150 mg twice per day of a 21 -day cycle and necitumumab is administered within the range of 400 to 1000 mg with dosages on 2 or 3 days of a 21-day cycle, alternatively with dosages one time a week or once every two weeks, preferably about 400 to 1000 mg on Day 1, Day 8, and Day 15 of a 21 -day cycle, more preferably about 600 to 900 mg on Day 1 and Day 8 of a 21 -day cycle, and most preferably about 800 mg on Day 1 and Day 8 of a 21-day cycle. Additional 21 -day cycles can be utilized as needed for treatment of the patient in need thereof.

The free base compound, abemaciclib, is preferred. However, it will be understood by the skilled reader that abemaciclib is capable of forming salts. LSN281354, a

monomesylate salt of abemaciclib, is an example of such a salt that can be used in preclinical studies. Abemaciclib can react with any of a number of inorganic and organic acids to form pharmaceutically acceptable acid addition salts. Such pharmaceutically acceptable acid addition salts and common methodology for preparing them are well known in the art. See, e.g., P. Stahl, et al, HANDBOOK OF PHARMACEUTICAL SALTS: PROPERTIES, SELECTION AND USE, (VCHA/Wiley-VCH, 2002); L.D. Bighley, S.M. Berge, D.C. Monkhouse, in "Encyclopedia of Pharmaceutical Technology' . Eds. J. Swarbrick and J.C. Boylan, Vol. 13, Marcel Dekker, Inc., New York, Basel, Hong Kong 1995, pp. 453-499; S.M. Berge, et al., "Pharmaceutical Salts", Journal of Pharmaceutical Sciences, Vol 66, No. 1, January 1977.

Abemaciclib, or a pharmaceutically acceptable salt thereof, and necitumumab are preferably formulated as pharmaceutical compositions administered by any route which makes the compounds bioavailable. The route of administration may be varied in any way, limited by the physical properties of the drugs and the convenience of the patient and the caregiver. Preferably, abemaciclib, or a pharmaceutically acceptable salt thereof, is administered orally. Alternatively, abemaciclib, or a pharmaceutically acceptable salt thereof, is formulated for parenteral administration, such as intravenous or subcutaneous administration. Preferably, necitumumab is formulated for parenteral administration, such as intravenous or subcutaneous administration, more preferably for intravenous administration. Such pharmaceutical compositions and processes for preparing same are well known in the art. (See, e.g. , Remington: The Science and Practice of Pharmacy (D.B. Troy, Editor, 21st Edition, Lippincott, Williams & Wilkins, 2006).

Dosage regimens may be adjusted to provide the optimum desired response (e.g. , a therapeutic response). Treatment dosages may be titrated using routine methods known to those of skill in the art to optimize safety and efficacy. Dosing schedules, for intravenous (i.v.) or non-intravenous administration, localized or systemic, or combinations thereof, will typically range from a single bolus dosage or continuous infusion to multiple administrations per day (e.g. , every 4-6 hours), or as indicated by the treating physician and the patient's condition.

Dosing in nonclinical murine models can guide initial clinical dosing for human patients. For necitumumab, the dose of 20 mg/kg twice weekly by intraperitoneal injection to mice showed a statistically significant decrease in tumor growth, while the decrease at 6 mg/kg was not statistically significant. From this information a minimum target serum concentration that would be needed for efficacy in cancer patients was determined. The serum concentrations in cancer patients were above this minimum target serum concentration after the 800 mg intravenous dose on day 1 and day 8 of a 21 -day cycle. For abemaciclib, an oral dose of 50 mg/kg once daily in mice achieved plasma concentrations that would yield efficacy in cancer patients. S. Tate et ah , Clin Cancer Res, July 15, 2014 20:3763. The plasma concentrations in cancer patients were above this target plasma concentration after repeated twice daily oral dosing of 200 mg by day 28 of a 28-day cycle.

As used herein, the phrase "in combination with" refers to the administration of abemaciclib, or a pharmaceutically acceptable salt thereof, with necitumumab

simultaneously. As used herein, the phrase "in combination with" also refers to the administration of abemaciclib, or a pharmaceutically acceptable salt thereof, with

necitumumab sequentially in any order. As used herein, the phrase "in combination with" also refers to the administration of abemaciclib, or a pharmaceutically acceptable salt thereof, with necitumumab in any combination thereof. Abemaciclib can be administered prior to administration of necitumumab. Abemaciclib can be administered at the same time as administration of necitumumab. Abemaciclib can be administered subsequent to

administration of necitumumab. Abemaciclib can be administered prior to, at the same time as, or subsequent to administration of necitumumab, or in some combination thereof.

Where necitumumab is administered at repeated intervals (e.g. during a standard course of treatment), abemaciclib can be administered prior to each administration of necitumumab. Where necitumumab is administered at repeated intervals (e.g. during a standard course of treatment), abemaciclib can be administered at the same time as each administration of necitumumab. Where necitumumab is administered at repeated intervals (e.g. during a standard course of treatment), abemaciclib can be administered subsequent to each administration of necitumumab. Where necitumumab is administered at repeated intervals (e.g. during a standard course of treatment), abemaciclib can be administered prior to, at the same time as, or subsequent to, each administration of necitumumab or some combination thereof. Where necitumumab is administered at repeated intervals (e.g. during a standard course of treatment), abemaciclib can be administered at different intervals in relation to therapy with necitumumab. Where necitumumab is administered at repeated intervals (e.g. during a standard course of treatment), abemaciclib can be administered in a single or series of dose(s) prior to, at any time during, or subsequent to the course of treatment with necitumumab. Where necitumumab is administered at repeated intervals (e.g. during a standard course of treatment), abemaciclib can be administered in a single dose prior to, at any time during, or subsequent to the course of treatment with necitumumab. Where necitumumab is administered at repeated intervals (e.g. during a standard course of treatment), abemaciclib can be administered in a single dose prior to the course of treatment with necitumumab. Where necitumumab is administered at repeated intervals (e.g. during a standard course of treatment), abemaciclib can be administered in a single dose at any time during the course of treatment with necitumumab. Where necitumumab is administered at repeated intervals (e.g. during a standard course of treatment), abemaciclib can be administered in a single dose subsequent to the course of treatment with necitumumab.

Where necitumumab is administered at repeated intervals (e.g. during a standard course of treatment), abemaciclib can be administered in a series of doses prior to the course of treatment with necitumumab. Where necitumumab is administered at repeated intervals (e.g. during a standard course of treatment), abemaciclib can be administered in a series of doses subsequent to the course of treatment with necitumumab. Where necitumumab is administered at repeated intervals (e.g. during a standard course of treatment), abemaciclib can be administered in a series of doses subsequent to the course of treatment with necitumumab.

The combination of necitumumab and abemaciclib may further be administered simultaneously, separately, or sequentially with gemcitabine and cisplatin.

The following examples and assays further illustrate the present invention.

The following assay results illustrate the unexpected improvement of the combination of necitumumab and abemaciclib in mouse xenograft models, patient derived xenograft models and cell line panel screenings.

The following assays and clinical study designs further illustrate the invention, but should not be construed to limit the scope of the invention in any way.

Abemaciclib can be made, for example, according to the disclosure in WO

2010/075074. Necitumumab can be made, for example, according to the disclosure in WO2005/090407.

Antitumor Effects of Abemaciclib in Combination with Necitumumab in an A549

Xenograft Mouse Model

To evaluate the efficacy of abemaciclib in combination with necitumumab, monotherapies and combinations are tested in an A549 Xenograft mouse model, which serves as a model for human NSCLC with an activating KRAS mutation.

The human NSCLC A549 line is grown in RPMI 1640 medium+10% fetal bovine serum. Sub-confluent cells are harvested with trypsin and rinsed twice with growth medium without serum. For subcutaneous tumors, growth is initiated by subcutaneous injection of 5 x 10⁶ cells in a 1: 1 mixture of Hank's balanced salt solution (HBSS) and Matrigel® (BD Bioscience, Franklin Lakes, NJ) in the rear flank of each subject animal. When mean tumors volumes reach approximately 150-200 mm in size, the animals are randomized by tumor size and body weight by randomization techniques well known in the art and placed into their respective treatment groups using 8 animals per group.

LSN2813542, a monomesylate salt of abemaciclib, is formulated on a weekly basis in 1% hydroxyethyl cellulose (HEC) in 25 mM phosphate buffer (PB) at a pH of 2 and stored at 4 °C. The monomesylate salt of abemaciclib is administered by oral gavage once daily for 28 days at doses of 50 mg/kg or 75 mg/kg using 0.2 mL /dose. Necitumumab is formulated in 0.9% injectable saline and is administered twice weekly for 4 weeks by intraperitoneal injection at a dose of 20 mg/kg. Animals in the combination therapy groups are given either 50 mg/kg or 75 mg/kg of the monomesylate salt of abemaciclib and 20 mg/kg of

necitumumab according to the schedules described above for monotherapy. The vehicle control group is given both the 1% HEC and 0.9% saline vehicles according to the schedules for the monomesylate salt of abemaciclib and necitumumab, respectively.

Tumor volume (V) is estimated by using the formula: V = 0.536L x W where L = larger of measured diameter and W = smaller of perpendicular diameter. Relative changes in tumor volume (%T/C) are calculated using the tumor volume measurements taken on the last day of dosing (study day 54) with the monomesylate salt of abemaciclib, whereas the baseline tumor volume is the volume recorded on or just prior to first day of dosing (study day 26). %T/C values are calculated using the formula %T/C = 100 x ΔΤ/AC, whereby T = mean tumor volume of the compound treated group, ΔΤ = mean tumor volume of the compound treated group minus the mean tumor volume on the baseline day, C = mean tumor volume of the control (vehicle) group, and AC = mean tumor volume of the control group minus the mean tumor volume on the baseline day. Tumor growth inhibition is observed in those instances where the calculated values for %T/C are less than 100% whereby greater inhibition results in smaller %T/C values. In experiments performed essentially as described, significant inhibition of the growth of the A549 xenograft tumors was observed in the monotherapies, including treatments with 50 mg/kg or 75 mg/kg of the monomesylate salt of abemaciclib and 20 mg/kg of necitumumab (Table 1). However, the greatest inhibition was seen in the group treated with 50 mg/kg of the monomesylate salt of abemaciclib in combination with 20 mg/kg of necitumumab.

Treatment of tumor-bearing mice with either 50 or 75 mg/kg of the monomesylate salt of abemaciclib resulted in a significant inhibition of the growth of the A549 xenograft tumors, whereby the %AT/C values determined at the end of the dosing period (day 54) were approximately 19% and 6%, respectively. These values were statistically significant from each other (p = 0.014), and from the vehicle-treated group (p < 0.001; Table 1), indicating that both doses were efficacious. Similarly monotherapy with necitumumab at 20 mg/kg resulted in significant inhibition in tumor growth. This treatment resulted in an observed %AT/C at the end of the treatment period (day 54) of 49% which was statistically significant when compared to the vehicle control group (p < 0.001; Table 1). Individual animal responses for the two monotherapy groups of the monomesylate salt of abemaciclib showed that treatment with 75 mg/kg as compared to 50 mg/kg resulted in a better DCR and a better response rate (RR). Specifically the DCR, which is the combined frequency of animals achieving SD, a PR or CR, observed for the 75 mg/kg group was 7/8 (87.5%) versus 4/8 (50%) for the group treated with 50 mg/kg. Likewise the RR, which is the combined frequency of animals achieving a either a PR or CR, was also better for the 75 mg/kg group as indicated by the response rates observed on day 54 of 4/8 (50%) and 0/8 (0%) for the 75 and 50 mg/kg groups, respectively. The DCR and RR for animals treated with necitumumab alone were 1/8 (12.5%) and 0/8 (0%), respectively.

Combining either 50 mg/kg or 75 mg/kg of the monomesylate salt of abemaciclib with necitumumab resulted in significant growth inhibition as compared to vehicle controls (p < 0.001). In particular, measurement of the A549 tumors at the end of the dosing period (day 54) for the combination group treated with 50 mg/kg the monomesylate salt of abemaciclib showed a near-complete growth inhibition with a %AT/C value of approximately 3% as compared to the %TA C values of 19% and 49% for monotherapy with the monomesylate salt of abemaciclib and necitumumab, respectively (Table 1). This effect observed for the combination was statistically significant as compared to either of the monotherapy groups (p < 0.002; Table 1). Combination of necitumumab with 50 mg/kg of the monomesylate salt of abemaciclib improved both the DCR and RR as compared to monotherapy resulting in a DCR of 8/8 (100%) for the combination versus 4/8 (50%) and 1/8 (12.5%) for monotherapy with the monomesylate salt of abemaciclib and necitumumab, respectively. Likewise the RR on day 54 for the combination was 2/8 (25%) versus 0/8 (0%) for both of the monotherapy groups.

Although significant antitumor efficacy was observed with the combination of necitumumab with 75 mg/kg the monomesylate salt of abemaciclib, this effect was not significantly different from the overall efficacy that was achieved with the monomesylate salt of abemaciclib alone. Assessment at the end of the dosing period (study day 54) showed that the combination of 75 mg/kg of the monomesylate salt of abemaciclib with necitumumab resulted in a %AT/C value of 4% which was significantly better than the %AT/C value of 49% (p < 0.001) for necitumumab, but not significantly better than the 6% (p = 0.695) observed for monotherapy with 75 mg/kg of the monomesylate salt of abemaciclib (Table 1). Accordingly the disease control rates (DCRs) for the combination group and the group receiving monotherapy with 75 mg/kg of the monomesylate salt of abemaciclib were also similar such that the DCRs observed on study day 54 were 7/8 (87.5%) and 8/8 (100%) for monotherapy and the combination therapy, respectively. The preclinical definitions of DCR, RR, SD, PR, and CR, used in this example may vary from the clinical definitions previously set forth. Table 1: Growth Inhibition of Subcutaneous A549 Xenografts by the Monomesylate Salt of Abemaciclib and Necitumumab

NA means not applicable. Antitumor Effects of Abemaciclib in Combination with Necitumumab in Patient

Derived Xenograft Mouse Models

To evaluate the efficacy of abemaciclib in combination with necitumumab, monotherapies and combinations are tested in patient-derived xenograft (PDX) tumor models of squamous non-small cell lung carcinoma (SqNSCLC).

These models are established from viable human tumor tissues that are serially passaged in immunocompromised female mice with limited frequency. Following essentially Migliardi et al., these PDXs are used to perform a large-scale in vivo screen to model inter-patient response heterogeneity with a "one animal per model per treatment" approach (l x l x l). Subcutaneous implants are performed unilaterally or bilaterally on the flank of animals using tumor fragments harvested from host animals implanted from as specific passage of the PDX tumor model. Animals are randomized into treatment groups and dosed when tumors volumes reach on or near 180-250 mm . Mice are enrolled on a rolling base and treated with necitumumab or the monomesylate salt (LSN 2813542) of abemaciclib in monotherapy and in combination without pre- selection on the basis of their genetic or proteomic, or growth latency/doubling time. The formulation and dosing regimen are listed in Table 2.

Table 2: Treatment Arms and Dosing Schedule

i.p. means intraperitoneal injection

p.o. means oral administration

s.c. means Subcutaneous injection

Combination treatment groups are given both compounds according to the schedule described for the monotherapy, while the vehicle treatment group receives a combination of both articles according to the schedule for each individual arm.

Tumor size and body weight are measured twice per week and tumor volume is calculated according to the formula:

Tumor volume = (a+b ) x 0.5 where "a" represents the largest of measured diameter and "b" the perpendicular tumor diameter. Relative changes in tumor volume (%AT/C) are calculated by dividing the absolute tumor volume taken at the end of the dosing period (on or near day 28), whereas the baseline tumor volume is the volume recorded on the first day of dosing (study day 0). %AT/C values are calculated using the formula:

%AT/C = AT/AC x 100

where T = mean tumor volume of the compound treated group, ΔΤ = mean tumor volume of the compound treated group minus the tumor volume on the baseline day, C = mean tumor volume of the control (vehicle) group, and AC = mean tumor volume of the control group minus the mean tumor volume on the baseline day. If ΔΤ was <0, then a tumor regression value is calculated instead of %AT/C whereby, %Regression = 100 x ΔΤ/Tini_ti_ai such that i_bi_al = the grand mean of the tumor volume for all the treatment groups.

The primary endpoint for tumor volume data, per animal, is best median response. To obtain this value, percent response is calculated at each measured time point for each treated animal relative to the mean of the vehicle controls. For tumor volumes above baseline, the percent response is Delta T/C, or ^ x 100%. For tumor volumes at or below baseline, the percent response is regression, or ^^{'* * " '} x 10¾%. Starting after two weeks of treatment, a moving window of three consecutive measurements is applied to the percent response vs. time data. The median percent response is recorded for each window, and the lowest of these medians is reported as the best median response.

To make pairwise comparisons between treatments, percent response results (Delta T/C or regression) is classified into one of three categories: PD (>10%), SD (-49% to +10%), or Regression (<= - 50%). For each pair of treatments, the number of models is counted for both treatments in the same category, one in a better category, or the other in a better category. McNemar's test is applied after combining Stable Disease and Regression into one category. McNemar's test is a categorical version of a paired t-test. The time to reach one doubling (event-free survival) is determined for each animal using linear interpolation of Log tumor volume vs time between consecutive measurements. A tumor that did not double is recorded as a right-censored value using the last day measured. Kaplan-Meier analysis is used to compare pairs of treatments and to plot the results. Hazard ratios are also calculated between each pair of treatments, and statistical significance is evaluated using Long Rank Test.

The results of this study (Table 3) based on the experiments performed essentially as described, suggest that the necitumumab/abemaciclib combination provides a benefit over either monotherapy as exemplified by the increased number of animals that achieved tumor regressions or stasis (15 vs 7, p=0.013 vs. necitumumab and 15 vs. 7, p=0.046 for

LSN2813542; McNemar's test). Median event-free survival (EFS) was also better for the combination (Table 4; mEFS = 34.3 vs 20.0 vs 14.2 days for the combination, the monomesylate salt of abemaciclib and necitumumab groups, respectively). In addition, the combination treatment was efficacious in 8 of 22 squamous NSCLC models that were intrinsically resistant (e.g. displayed PD) to gemcitabine/cisplatin (Table 5).

Table 3: Antitumor Response in Mice Bearing SqNSCLC PDX Tumors Treated with Single Agent Necitumumab or the Monomesylate Salt (LSN 2813542) of Abemaciclib or Necitumumab/the Monomesylate Salt (LSN 2813542) of Abemaciclib Combination

Necitumumab

Necitumumab LSN2813542

+LSN2813542 best best best

Response Response Response

Model dT/C% or dT/C% or dT/C% or

category category category regr% regr% regr%

LUN025 87.7 PD 70.1 PD -20.6 SD

LUN026 54.6 PD 35.3 PD -79.9 Regression

LUN035 168.5 PD 88.3 PD 18.5 PD

LUN038 67.5 PD 9.9 SD -3.8 SD

LUN051 -100.0 Regression 40.6 PD -100.0 Regression LUN112 -26.1 SD 5.1 SD 57.7 PD

LUN139 26.1 PD 60.2 PD -48.7 SD

LUN143 27.5 PD 24.4 PD -42.7 SD

LUN148 122.5 PD 59.4 PD 66.0 PD

LUN150 -66.5 Regression 18.6 PD -60.8 Regression

LUN167 -22.7 SD -11.5 SD 29.4 PD

LUN178 56.1 PD 211.8 PD 52.7 PD

LUN186 70.3 PD 120.2 PD -2.5 SD

LUN210 85.5 PD 19.6 PD 27.8 PD

LUN216 37.5 PD 102.4 PD 66.7 PD

LUN262 133.9 PD 49.3 PD 60.3 PD

LUN268 27.8 PD -0.5 SD -4.5 SD

LUN272 40.4 PD 12.0 PD 22.2 PD

LUN278 -75.5 Regression 80.0 PD -59.7 Regression

LUN296 68.0 PD 34.7 PD 49.5 PD

LXFE 1066 74.9 PD 39.7 PD 37.2 PD

LXFE 1422 91.6 PD 58.6 PD 186.7 PD

LXFE 211 60.0 PD -1.7 SD -58.9 Regression

LXFE 2159 45.3 PD 25.3 PD -34.5 SD

LXFE 2162 38.7 PD 61.6 PD 10.1 PD

LXFE 2190 64.2 PD 116.5 PD 77.7 PD

LXFE 2198 47.2 PD 58.1 PD 79.9 PD

LXFE 2220 199.0 PD 45.6 PD 31.3 PD

LXFE 2226 144.9 PD 35.8 PD 25.8 PD

LXFE 2232 56.4 PD 79.4 PD 51.2 PD

LXFE 2257 79.6 PD 50.5 PD 88.3 PD

LXFE 2276 85.8 PD -0.2 SD 18.0 PD

LXFE 397 340.6 PD 74.7 PD 165.1 PD

LXFE 409 210.3 PD 0.2 SD 13.5 PD

LXFE 470 275.0 PD 32.7 PD 25.7 PD

LXFE 597 -18.4 SD 72.6 PD -58.8 Regression

LXFE 646 111.3 PD 16.8 PD -100.0 Regression

LXFE 690 -100.0 Regression 15.9 PD NE NE

LXFE 772 107.5 PD 127.0 PD 60.3 PD

LXFE 937 58.5 PD 104.0 PD -37.5 SD

NE means that the animals did not meet our pre- specified criteria for having sufficient data to reliably calculate an efficacy result. Table 4: Event-Free Survival in Mice Bearing SqNSCLC PDX Tumors Treated with Single Agent Necitumumab or the Monomesylate Salt (LSN 2813542) of Abemaciclib or Necitumumab/the Monomesylate Salt (LSN 2813542) of Abemaciclib Combination

Table 5: Antitumor Response in Mice Bearing SqNSCLC PDX Tumors Treated with

A p=0.042 and p=0.006 vs necitumumab and

LSN2813542 monotherapy, respectively (Long Rank Test)

Gemcitabine/Cisplatin or Necitumumab/the Monomesylate Salt (LSN 2813542) of

Abemaciclib Combination

Necitumumab + LSN2813542 Gem/Cis

Model best dT/C% Response best dT/C% Response

or regr% category or regr% category

LUN148 66.0 PD -100.0 Regression

LUN210 27.8 PD -100.0 Regression

LUN026 -79.9 Regression -86.9 Regression

LUN051 -100.0 Regression -78.5 Regression

LUN112 57.7 PD -77.3 Regression

LUN178 52.7 PD -40.1 SD

LXFE 409 13.5 PD -40.0 SD

LUN272 22.2 PD -34.3 SD

LUN035 18.5 PD -33.3 SD

LUN296 49.5 PD -7.9 SD

LUN268 -4.5 SD -4.9 SD LUN150 -60.8 Regression -4.2 SD

LUN278 -59.7 Regression 1.7 SD

LXFE 211 -58.9 Regression 1.8 SD

LXFE 1422 186.7 PD 3.8 SD

LUN216 66.7 PD 7.2 SD

LXFE 2162 10.1 PD 12.9 PD

LXFE 2220 31.3 PD 13.5 PD

LUN262 60.3 PD 17.1 PD

LXFE 2159 -34.5 SD 24.2 PD

LUN025 -20.6 SD 26.3 PD

LXFE 1066 37.2 PD 27.8 PD

LUN143 -42.7 SD 28.1 PD

LXFE 2232 51.2 PD 30.1 PD

LUN186 -2.5 SD 34.8 PD

LXFE 690 NE NE 36.9 PD

LXFE 2226 25.8 PD 38.9 PD

LUN139 -48.7 SD 44.2 PD

LXFE 2257 88.3 PD 66.2 PD

LXFE 597 -58.8 Regression 84.1 PD

LXFE 397 165.1 PD 99.7 PD

LXFE 937 -37.5 SD 100.2 PD

LXFE 646 -100.0 Regression 109.7 PD

LXFE 2190 77.7 PD 110.7 PD

LXFE 2198 79.9 PD 117.9 PD

LXFE 470 25.7 PD 130.0 PD

LXFE 772 60.3 PD 177.2 PD

LXFE 2276 18.0 PD 197.0 PD

LUN038 -3.8 SD NE NE

LUN167 29.4 PD NE NE

NE means that the animals did not meet our pre- specified criteria for having sufficient data to reliably calculate an efficacy result.

According, these data suggest that the necitumumab/abemaciclib combination provides a benefit over either monotherapy as exemplified by increased number of animals that achieved tumor regressions or stasis. Antitumor Effects of Abemaciclib in Combination with Necitumumab Across a Diverse Panel of Cancer Cell Lines

To evaluate the efficacy of abemaciclib in combination with necitumumab, monotherapies and combinations of the monomesylate salt (LSN2813542) of abemaciclib and necitumumab are tested across a panel of cancer cell lines of diverse genetics and histology types including lung, colorectal, stomach, skin, ovary, prostate, and pancreas cancer.

The monomesylate salt (LSN2813542) of abemaciclib is tested as a single agent across a large panel of cancer cell lines to identify the determinants of response. Inhibition of cell growth, after a duration of two cell doublings calculated for the vehicle control (DMSO), is measured using CellTiterGlo® (Promega Corp) and statistical methods are used to identify genes whose expression significantly correlates with resistance to abemaciclib.

In the analysis of the experiments performed essentially as described, it was shown that expression levels of EGFR correlate with LSN 2813542 IC₅₀ (FDR < 0.000002 from testing expression of 17331 genes), i.e. EGFR expression predicts resistance to abemaciclib. These data suggested that the combination of an EGFR inhibitor, such as necitumumab, could sensitize cancer cells to abemaciclib by targeting a resistance factor. The

monomesylate salt (LSN2813542) of abemaciclib and necitumumab are tested in

combination and their efficacy is compared to the single agent data across a panel of 497 cancer cell lines of diverse genetics and histology type. In these studies necitumumab is used at a fixed dose of 20 μg/mL and LSN2813542 is tested in a 10 point titration. An IC₅₀ is calculated from these data for the combination and the single agent LSN2813542. Table 6 shows the 191 cell lines where the IC50 of abemaciclib in the presence of 20 μg/mL necitumumab is less than 2 uM. These cell lines represent the cancer types that are predicted to most likely to benefit from this combination. Table 7 shows the 32 cell lines where the combination unexpectedly showed the best evidence of better activity than the single agents alone at concentrations of abemaciclib that are clinically achievable. In Table 7 the 32 cell lines where (1) the ratio of the combination IC₅₀ to LSN2813542 single agent IC₅₀ is 0.75 or better, (2) the necitumumab activity in the presence of the lowest dose of LSN2813542 (InM) was less than 35% and (3) the absolute IC₅₀ of the combination was less than ΙμΜ are listed. This list includes lung, colorectal, stomach, skin, ovary, prostate, and pancreas cancer.

All cell lines are obtained from commercial sources and are cultured in conditions recommended by vendors (ATCC, JCRB, SIBS, ECACC, RIKEN, CLS, LONZA, AllCells, ChemPartner, CASTCC, HSRRB, DSMZ, ICLC, SNU). Cell line histology and site of origin annotation is derived from the vendor or the COSMIC cancer cell line database (www.cancer.sanger.ac.uk). Prior to use, cell lines are tested for mycoplasma. Cell line authenticity is confirmed by STR-based DNA finger printing and multiplex PCR (IDEXX- Radil). For mycoplasma-free cultures with authentic STR fingerprints, growth curves are determined to establish average population doubling time in the absence of drug treatment for each cell line. Cell density is optimized to ensure robust, logarithmic cell growth for the duration of compound exposure.

Cell proliferation Assay: Cells are recovered from frozen stocks and cultured for 1-2 passages in T-75 or T-175 cell culture flasks. The day before profiling, cells are seeded in complete media in 384-well white-walled clear bottom microtiter plates at the predetermined optimal density for each cell line. Sixteen hours after plating, compound/antibody is added. CellTiter-Glo® (Promega Corporation, Madison, WI) reagents are prepared according to the manufacturer's protocol. Two doubling times after compound addition, CellTiter-Glo® reagent is added to each well. Plates are incubated at room temperature for 10 minutes before recording with a luminescence plate reader (EnSpire® Multimode Plate Reader and En Vision® Multilabel Plate Reader, Perkin Elmer, 940 Winter St., Waltham, Massachusetts 02451). IC₅₀ is determined by fitting a standard 4-parameter logistic model to the

luminescent measurements from 10 point titrations.

Statistical Methods: Regression and ANOVA models are applied to test the significance of the association between genomic features and efficacy. LoglO Likelihood Ratio (LOD) and P-value are computed by comparing each gene model to the null model. False discovery rate (FDR) is computed using Benjamini Hochberg method.

Genomic Data: Gene mutation, copy number and expression data is compiled from public domain data sets from COSMIC (www.cancer.sanger.ac.uk) and CCLE

(w w w .bro adinstitue . org/ccle/home) .

Table 6: IC50 Ratios of Necitumumab and the Monomesylate Salt (LSN 2813542) of Abemaciclib as Compared to the Monomesylate Salt (LSN 2813542) of Abemaciclib

Monotherapy in Various Tumor Types

Cell LSN2813542 Necitumumab LSN2813542 IC50 Cell Line Source

Line IC50 (2(^g/mL) + (combo):

% inhibition Necitumumab IC50

(2(^g/mL) (single)

IC50

NCI- 3.077 46.400 0.001 0.000 Lung

H292

DoTc2- 0.378 57.250 0.001 0.003 Cervix

4510

NCI- 2.234 65.803 0.001 0.000 Lung

HI 623 ( Adenoc arcinoma)

NCI- 1.699 60.404 0.001 0.001 Lung

H2073 (Adenoc arcinoma)

RMG-I 1.356 57.219 0.001 0.001 Ovary

SW1463 0.069 41.838 0.012 0.173 Large Intestine

NCI- 2.075 41.275 0.024 0.012 Lung

H1648 (Adenoc arcinoma)

JeKo-1 0.040 -21.549 0.048 1.190 Haematopoietic and Lymphoid Tissue

(Blood NHL)

SK- 0.958 36.687 0.080 0.084 Lung (Squamous)

MES-1

NCI- 0.764 35.200 0.100 0.131 Lung (Squamous)

H1869

NCI- 0.169 52.885 0.102 0.604 Lung

H1975 (Adenoc arcinoma) MCFlOa 0.474 38.626 0.112 0.237 Breast (Normal)

HCC827 1.516 31.987 0.113 0.074 Lung

(Adenoc arcinoma)

Kasumi- 0.160 -3.540 0.122 0.765 Haematopoietic

6 and Lymphoid

Tissue (Blood AML)

YCC-6 1.114 36.580 0.142 0.127 Stomach

Z-138 0.086 -10.270 0.144 1.669 Haematopoietic and Lymphoid Tissue (Blood NHL)

NCI- 0.345 13.188 0.147 0.426 Lung (SCLC)

H2286

IA-LM 0.215 -4.158 0.184 0.855 Lung

(Adenoc arcinoma)

CHP- 0.304 3.972 0.189 0.623 Autonomic

212 Ganglia

(Neuroblastoma)

NCI- 0.571 17.500 0.190 0.333 Lung

H358 (Adenoc arcinoma)

LoVo 0.553 8.249 0.192 0.347 Large Intestine

Reh 0.248 -12.388 0.198 0.799 Haematopoietic and Lymphoid Tissue (Blood ALL)

DMS 53 0.143 43.604 0.226 1.577 Lung (SCLC)

D283 0.278 9.900 0.230 0.827 CNS

Med Medulloblastoma

BCP-1 0.281 8.124 0.231 0.823 Haematopoietic and Lymphoid Tissue (Blood ALL)

Caki-2 2.313 33.473 0.245 0.106 Kidney (Renal)

Pfeiffer 0.290 -16.690 0.262 0.902 Haematopoietic and Lymphoid Tissue (Blood NHL)

PvEC-1 0.283 -1.713 0.279 0.984 Haematopoietic and Lymphoid Tissue (Blood NHL) LIXC- 0.247 -0.103 0.289 1.173 Liver

012

TALL- 1 0.227 -2.583 0.307 1.357 Haematopoietic and Lymphoid Tissue (Blood ALL)

CCRF- 0.500 -9.564 0.313 0.625 Haematopoietic CEM and Lymphoid

Tissue (Blood ALL)

Daudi 0.288 -9.366 0.322 1.118 Haematopoietic and Lymphoid Tissue (Blood NHL)

F-36P 0.335 9.509 0.329 0.981 Haematopoietic and Lymphoid Tissue (Blood RAEB)

TT 0.277 4.057 0.335 1.209 Thyroid

(Squamous)

HuGl-N 0.859 8.073 0.366 0.427 Stomach

NCI- 0.666 12.356 0.424 0.637 Lung (NSCLC) HI 944

KE-97 0.512 -6.100 0.430 0.840 Stomach

LUDLU 0.835 8.413 0.446 0.534 Lung (Squamous) -1

QGP-1 0.878 3.490 0.464 0.528 Pancreas

(Somatostatinoma)

MMAC- 0.494 6.883 0.465 0.940 Skin (Melanoma) SF

GA- 0.485 -1.200 0.480 0.990 Haematopoietic 10(Clon and Lymphoid e4) Tissue (Blood

NHL)

ACHN 0.324 6.566 0.518 1.597 Kidney (Renal)

P30/OH 0.496 1.399 0.519 1.045 Haematopoietic K and Lymphoid

Tissue (Blood ALL)

CA46 0.620 -6.500 0.520 0.839 Haematopoietic and Lymphoid Tissue (Blood NHL)

NCI- 0.742 2.661 0.523 0.705 Lung

H1792 (Adenoc arcinoma)

NCI- 1.665 4.200 0.540 0.324 Lung

H2122 (Adenoc arcinoma)

NCI- 4.081 16.978 0.561 0.138 Lung

H1734 (Adenoc arcinoma)

KASUM 0.522 0.128 0.602 1.154 Haematopoietic

11 and Lymphoid

Tissue (Blood AML)

PA-1 0.534 -7.796 0.612 1.144 Ovary

YCC-11 0.795 -0.289 0.625 0.786 Stomach

G-401 0.365 11.500 0.640 1.756 Kidney

(Rhabdoid)

RL95-2 1.224 31.900 0.640 0.523 Endometrium

NCI- 1.122 4.854 0.649 0.578 Lung

H1355 (Adenoc arcinoma)

22RV1 1.396 7.591 0.657 0.471 Prostate

(Adenoc arcinoma)

D341Me 0.522 -7.817 0.667 1.278 CNS

d (MeduUoblastoma)

Nb2-l l 0.501 -3.099 0.685 1.367 Haematopoietic and Lymphoid Tissue (Blood Lymphoma)

NCI- 2.765 6.286 0.685 0.248 Lung

H1395 (Adenoc arcinoma)

YCC-10 0.622 -0.997 0.697 1.120 Stomach

NH-6 0.673 11.108 0.720 1.070 Autonomic

Ganglia

(Neuroblastoma)

RKN 0.325 -11.932 0.736 2.265 Ovary (Soft

Tissue)

RS4;11 0.831 8.500 0.740 0.890 Haematopoietic and Lymphoid Tissue (Blood ALL)

NCI- 1.265 0.359 0.749 0.592 Lung

H2405 (Adenoc arcinoma)

VA-ES- 2.884 29.700 0.760 0.264 Bone (Epithelioid BJ Sarcoma)

Gp2d 1.089 12.361 0.776 0.713 Large Intestine

CV-1 1.365 19.702 0.778 0.570 Kidney (NS)

Hs746T 1.086 -0.700 0.790 0.727 Stomach

NCI- 0.838 -4.332 0.796 0.949 Lung (SCLC) H524

SNU-C1 0.901 -1.383 0.799 0.887 Large Intestine

OCUM- 0.786 -13.467 0.802 1.021 Stomach

1

NCI- 2.196 5.925 0.819 0.373 Lung

H2085 (Adenoc arcinoma)

C2BBel 2.277 29.628 0.825 0.362 Large Intestine

Loucy 0.898 -9.737 0.829 0.923 Haematopoietic and Lymphoid Tissue (Blood ALL)

OVMA 1.935 34.000 0.830 0.429 Ovary

NA

BxPC-3 0.971 8.332 0.845 0.870 Pancreas

(Adenoc arcinoma)

HCCC- 0.987 16.126 0.846 0.857 Liver

9810

Capan-1 1.170 7.799 0.855 0.731 Pancreas

(Adenoc arcinoma)

GCT 1.344 -8.100 0.870 0.647 Soft Tissue

NCI- 1.179 -2.635 0.877 0.744 Lung

H650 (Adenoc arcinoma)

KG-1 0.636 -8.823 0.882 1.385 Haematopoietic and Lymphoid Tissue (Blood AML)

HCC193 0.848 -6.118 0.886 1.045 Breast (TNBC) 7

LIXC- 1.916 21.293 0.900 0.470 Liver

011

NCCIT 0.773 4.400 0.900 1.164 Testis

A-431 2.334 21.500 0.920 0.394 Skin (Melanoma)

NCI- 2.021 9.990 0.941 0.465 Pleura

H2052 (Mesothelioma)

NCI- 0.798 -7.739 0.943 1.182 Lung (SCLC) H209 MLMA 1.146 3.320 0.952 0.831 Haematopoietic and Lymphoid Tissue (Blood NHL)

A2780 1.026 2.342 0.986 0.961 Ovary

HMV-II 1.023 -9.689 0.993 0.970 Skin (Melanoma)

NCI- 1.065 3.583 1.007 0.945 Lung (SCLC)

H2171

THP-1 1.230 -2.395 1.008 0.820 Haematopoietic and Lymphoid Tissue (Blood AML)

A549 1.355 -2.792 1.016 0.750 Lung (NSCLC)

MDA- 1.172 -4.332 1.020 0.871 Breast (HER2)

MB-453

NCI- 1.603 21.443 1.029 0.642 Lung

H1573 (Adenoc arcinoma)

SW620 1.031 3.937 1.033 1.002 Large Intestine

SK- 1.541 1.462 1.040 0.675 Skin (Melanoma)

MEL-3

Toledo 1.160 3.600 1.040 0.896 Haematopoietic and Lymphoid Tissue (Blood NHL)

U251 1.168 -7.359 1.050 0.899 CNS

LNCaP 0.875 0.803 1.055 1.206 Prostate clone (Adenoc arcinoma)

FGC

T98G 1.209 3.621 1.058 0.875 CNS

(Glioblastoma)

NCI- 4.433 21.971 1.073 0.242 Lung

H2126 (Adenoc arcinoma)

T.T 1.208 8.168 1.077 0.892 Thyroid

(Squamous)

COR- 0.852 -9.610 1.087 1.276 Lung (SCLC)

L279

Hela 1.137 -12.328 1.087 0.956 Cervix

SK-LU- 0.996 0.773 1.094 1.098 Lung

1 (Adenoc arcinoma)

NUGC- 1.927 29.704 1.142 0.593 Stomach

3 HMCB 0.999 -2.536 1.146 1.147 Skin (Melanoma)

BEL740 1.289 71.058 1.157 0.898 Liver

5

RPMI66 0.786 8.700 1.160 1.476 Haematopoietic 66 and Lymphoid

Tissue (Blood Hodgkin

Lymphoma)

769-P 1.693 12.800 1.170 0.691 Kidney (Renal)

Hs940.T 0.688 -3.900 1.170 1.702 Skin (Melanoma)

NCI- 1.715 -11.315 1.174 0.684 Lung (SCLC) H2107

RPMI 1.761 8.632 1.186 0.673 Haematopoietic

8226 and Lymphoid

Tissue (Blood Multiple

Myeloma)

A-704 1.302 6.857 1.187 0.912 Kidney (Renal)

HGC-27 1.552 -4.700 1.190 0.767 Stomach

RT112- 1.548 10.066 1.233 0.796 Urinary Tract (NS) 84

Caov-3 1.430 4.432 1.251 0.875 Ovary

HT1376 5.273 21.104 1.259 0.239 Urinary Tract

(Transitional Cell Carcinoma)

NCI- 1.768 -3.966 1.264 0.715 Lung

H1703 (Adenoc arcinoma)

NCI- 1.932 7.363 1.271 0.658 Lung (Squamous) H2170

PF-382 0.929 -9.697 1.276 1.374 Haematopoietic and Lymphoid Tissue (Blood ALL)

RD-ES 1.206 2.499 1.279 1.060 Bone (Ewings

Sarcoma)

NCI- 1.279 3.441 1.292 1.010 Lung (SCLC) H2227

RERF- 2.118 1.021 1.302 0.615 Lung

LC-MS (Adenoc arcinoma)

HFF-1 1.611 4.579 1.303 0.809 Skin (Melanoma

Normal) NCI- 2.367 -6.000 1.310 0.554 Lung (SCLC) H1876

SNU-1 0.969 -6.151 1.318 1.360 Stomach

Hs852.T 1.561 5.057 1.331 0.853 Skin (Melanoma)

DMS 79 1.973 16.826 1.347 0.683 Lung (SCLC)

HLF 1.285 -12.437 1.356 1.055 Liver

COLO- 1.115 2.913 1.362 1.221 Large Intestine 205

SK-N- 1.145 -4.540 1.370 1.196 Autonomic AS Ganglia

(Neuroblastoma)

MDA- 1.766 -3.900 1.370 0.776 Breast (TNBC) MB-157

HLE 1.737 3.207 1.381 0.795 Liver

MOLT- 1.701 -4.933 1.393 0.819 Haematopoietic 4 and Lymphoid

Tissue (Blood ALL)

CHL-1 1.001 -7.705 1.395 1.394 Skin (Melanoma)

NCI- 2.086 -1.300 1.410 0.676 Lung (SCLC) H1048

SK- 2.779 -8.400 1.420 0.511 Skin (Melanoma) MEL-5

HuVEC 1.480 -19.910 1.425 0.962 Umbilical Vein

Endothelial Cell (Normal)

Hs 294T 2.355 -5.081 1.426 0.605 Skin (Melanoma)

HCC156 1.446 -13.720 1.434 0.992 Breast (HER2) 9

786-0 1.816 15.646 1.446 0.797 Kidney (Renal)

MX-1 1.763 -1.266 1.460 0.828 Breast (TNBC)

SK- 1.687 -7.909 1.465 0.869 Skin (Melanoma)

MEL-24

ATN-1 0.933 0.159 1.465 1.571 Haematopoietic and Lymphoid Tissue (Blood ALL)

SW1417 2.616 5.446 1.473 0.563 Large Intestine

Ramos.2 1.420 -3.700 1.480 1.042 Haematopoietic G6.4C10 and Lymphoid

Tissue (Blood NHL)

LS411N 2.322 -3.785 1.491 0.642 Large Intestine

TOV- 2.953 14.900 1.510 0.511 Ovary

21G

HL-60 1.623 -0.765 1.550 0.955 Haematopoietic and Lymphoid Tissue (Blood AML)

C8166 1.823 4.480 1.555 0.853 Haematopoietic and Lymphoid Tissue (Blood ALL)

NCI- 1.026 1.700 1.560 1.521 Lung (SCLC) H82

HCC114 1.462 14.139 1.560 1.067 Breast (TNBC)

3

UACC- 1.561 1.935 1.567 1.004 Breast

812

A101D 1.547 -9.954 1.580 1.022 Skin (Melanoma)

Caki-1 1.027 15.521 1.584 1.542 Kidney (Renal)

NCI- 2.057 29.514 1.595 0.775 Lung

H1666 (Adenoc arcinoma)

C3A 1.466 4.937 1.609 1.097 Liver

DU145 4.594 19.806 1.609 0.350 Prostate

(Adenoc arcinoma)

AN3 CA 1.610 3.616 1.610 1.000 Endometrium

HCT116 1.413 3.100 1.610 1.139 Large Intestine

Panc03. 1.383 1.200 1.620 1.171 Pancreas

27 (Adenoc arcinoma)

SK- 1.565 -10.300 1.640 1.048 Lung (Squamous) MES-1

LIXC- 1.215 21.235 1.655 1.362 Liver

002

NB 16 1.451 -7.895 1.662 1.145 Autonomic

Ganglia

(Neuroblastoma)

MGC80- 2.327 -0.100 1.690 0.726 Stomach

3

RT4 2.247 13.766 1.692 0.753 Urinary Tract

Transitional Cell (Carcinoma) G-361 1.347 16.200 1.700 1.262 Skin (Melanoma)

NCI- 1.876 -0.504 1.717 0.915 Lung (SCLC)

H510A

NAMA 1.899 -8.114 1.726 0.909 Haematopoietic

LWA and Lymphoid

Tissue (Blood NHL)

TGBC1 1.271 5.233 1.744 1.372 Biliary Tract

TKB

KLE 1.796 13.374 1.750 0.974 Endometrium

DB 1.372 -0.863 1.759 1.282 Haematopoietic and Lymphoid Tissue (Blood NHL)

CW-2 1.422 4.994 1.761 1.238 Large Intestine

CAL 27 1.669 -0.454 1.779 1.066 Upper

Aerodigestive Tract (Squamous)

MDCK 1.663 -2.600 1.810 1.088 Kidney (NS)

FaDu 2.549 20.816 1.814 0.711 Upper

Aerodigestive Tract (Squamous)

U266 1.674 1.176 1.839 1.098 Haematopoietic and Lymphoid Tissue (Blood Multiple

Myeloma)

NCI- 2.085 3.500 1.840 0.882 Lung

H2023 (Adenoc arcinoma)

SW-13 2.101 -12.796 1.847 0.879 Adrenal Gland

SNU- 3.885 8.073 1.855 0.477 Stomach

638

EB 1 1.738 -0.737 1.861 1.071 Haematopoietic and Lymphoid Tissue (Blood NHL)

T24 2.230 -9.251 1.874 0.840 Urinary Tract

Transitional Cell (Carcinoma)

KATO 1.804 -20.211 1.895 1.051 Stomach

III ARH-77 2.530 1.480 1.902 0.752 Haematopoietic and Lymphoid Tissue (Blood Multiple Myeloma)

MM1S 1.478 -9.897 1.917 1.297 Haematopoietic and Lymphoid Tissue (Blood Multiple Myeloma)

NIH:OV 4.033 8.360 1.918 0.476 Ovary

CAR-3

NCI- 2.724 -10.935 1.918 0.704 Lung (SCLC)

H2029

SK- 2.808 -0.300 1.920 0.684 Kidney (NS)

NEP-1

Pane 2.302 -8.820 1.929 0.838 Pancreas (NS)

04.03

MKN- 2.768 11.824 1.996 0.721 Stomach

45

Table 7: IC50 Ratios of Necitumumab and the Monomesylate Salt (LSN 2813542) of Abemaciclib as Compared to the Monomesylate Salt (LSN 2813542) of Abemaciclib

Monotherapy in Various Tumor Types

Cell LSN2813542 Necitumumab LSN2813542 + IC50 Cell Line Source Line IC50 (2(^g/mL) Necitumumab (combo)

% inhibition (2(^g/mL)

IC50 IC50

(single)

HCC8 1.516 31.987 0.113 0.074 Lung

27 ( Adenoc arcinoma)

Caki- 2.313 33.473 0.245 0.106 Kidney

2

NCI- 4.081 16.978 0.561 0.138 Lung

H1734 (Adenoc arcinoma) NCI- 2.765 6.286 0.685 0.248 Lung

H1395 (Adenoc arcinoma)

VA- 2.884 29.700 0.760 0.264 Bone (Epithelioid ES-BJ sarcoma)

NCI- 1.665 4.200 0.540 0.324 Lung

H2122 (Adenoc arcinoma)

NCI- 0.571 17.500 0.190 0.333 Lung

H358 (Adenoc arcinoma)

LoVo 0.553 8.249 0.192 0.347 Large Intestine

2.277 29.628 0.825 0.362 Large Intestine

C2BB

el

NCI- 2.196 5.925 0.819 0.373 Lung

H2085 (Adenoc arcinoma)

A-431 2.334 21.500 0.920 0.394 Skin (Melanoma)

NCI- 0.345 13.188 0.147 0.426 Lung (SCLC) H2286

0.859 8.073 0.366 0.427 Stomach

HuGl- N

1.935 34.000 0.830 0.429 Ovary

OVM

ANA

NCI- 2.021 9.990 0.941 0.465 Pleura

H2052 (Mesothelioma)

LIXC- 1.916 21.293 0.900 0.470 Liver

011

22RV1 1.396 7.591 0.657 0.471 Prostate

(Adenoc arcinoma)

RL95- 1.224 31.900 0.640 0.523 Endometrium 2

QGP-1 0.878 3.490 0.464 0.528 Pancreas

(S omato s tatinoma)

LUDL 0.835 8.413 0.446 0.534 Lung (Squamous) U-l

CV-1 1.365 19.702 0.778 0.570 Kidney

NCI- 1.122 4.854 0.649 0.578 Lung

H1355 (Adenoc arcinoma)

NCI- 1.265 0.359 0.749 0.592 Lung

H2405 (Adenoc arcinoma)

CHP- 0.304 3.972 0.189 0.623 Autonomic 212 Ganglia (Neuroblastoma)

CCRF- 0.500 -9.564 0.313 0.625 Haematopoietic

CEM and Lymphoid

Tissue

NCI- 0.666 12.356 0.424 0.637 Lung (NSCLC)

HI 944

GCT 1.344 -8.100 0.870 0.647 Soft Tissue

NCI- 0.742 2.661 0.523 0.705 Lung

H1792 ( Adenoc arcinoma)

Gp2d 1.089 12.361 0.776 0.713 Large Intestine

Hs 1.086 -0.700 0.790 0.727 Stomach

746T

Capan- 1.170 7.799 0.855 0.731 Pancreas

1 (Adenoc arcinoma)

NCI- 1.179 -2.635 0.877 0.744 Lung

H650 (Adenoc arcinoma)

Accordingly, the data suggests that the combination of necitumumab and abemaciclib provides better activity than the single agents at concentrations of abemaciclib that are clinically achievable in numerous cancers including but not limited to lung, colorectal, stomach, skin, ovary, prostate, and pancreas cancer.

A Study of Abemaciclib in Combination with Necitumumab

for Patients with Stage IV NSCLC

Clinical Study Design

A single-arm, multicenter Phase lb study (hereinafter "Study") with an expansion cohort to investigate necitumumab in combination with abemaciclib in approximately 70 patients with Stage IV NSCLC (American Joint Committee on Cancer Staging Manual, 7th edition). The study consists of 2 parts:

• Part A: Dose-escalation part with increasing doses of abemaciclib (100 mg, 150 mg, or 200 mg every 12 hours [Q12H] on Days 1 to 21) to determine a recommended dose range for abemaciclib that may be safely administered in combination with a fixed regimen of necitumumab 800 mg on Days 1 and 8 every 21 days in patients with Stage IV NSCLC.

• Part B (expansion cohort): Dose confirmation of abemaciclib in combination with a fixed regimen of necitumumab 800 mg on Days 1 and 8 every 21 days and exploration of clinical antitumor activity.

Approximately 50 patients will be enrolled in Part B: approximately 25 patients with squamous histology and approximately 25 patients with nonsquamous histology.

Patients will be treated until PD, toxicity requiring cessation, protocol

noncompliance, or withdrawal of consent. Patients who are on study therapy at study completion may continue to receive study therapy in the extension phase until they meet the discontinuation criteria. Study completion is expected to be approximately 5 months after the last patient has been enrolled.

Study Objectives

The primary objectives of the study are (i) to determine the dose-limiting toxicity

(DLT) of abemaciclib at doses up to 200 mg when combined with necitumumab 800 mg, in patients with Stage IV NSCLC as measured by the number of patients with a DLT in Cycle 1 and (ii) to evaluate the efficacy of necitumumab in combination with abemaciclib in terms of PFS rate at 3 months in patients with Stage IV NSCLC.

The secondary objectives of the study are (i) to investigate the safety profile as assessed by clinical and laboratory significant events of necitumumab in combination with abemaciclib; (ii) to determine the overall response rate (ORR); (iii) to determine the pharmacokinetics (PK) of necitumumab and abemaciclib; (iv) to determine the

immunogenicity of necitumumab; and (v) to demonstrate the safety, efficacy, and feasibility of necitumumab in combination with abemaciclib at the recommended dose.

The exploratory objective of the study is to correlate biomarkers with clinical outcomes, including, but not limited to, KRAS mutation assessment, EGFR protein expression, and/or other biomarkers associated with the disease pathobiology, the cell cycle, EGFR pathway, and/or the mechanism of action of the therapeutic molecules.

Trial Drugs:

Abemaciclib is an orally available CDK4 and CDK6 inhibitor supplied as capsules

(50 mg/capsule) for oral administration. Abemaciclib may also be provided in a tablet form (50 mg/tablet) for oral administration. Necitumumab is a sterile, preservative-free, I.V. infusion supplied at a final concentration of 16 mg/mL (800 mg/50 mL) contained in single- use vials, in a formulation of 10 mM citrate, 40 mM sodium chloride, 133 mM glycine, 50 mM mannitol, 0.01% polysorbate-(TWEEN®)-80, pH 6.0.

Claims

WE CLAIM:

1. A method of treating cancer in a patient, wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma, comprising administering to the patient in need of such a treatment an effective amount of necitumumab in combination with an effective about of abemaciclib, or a pharmaceutically acceptable salt thereof.

2. A method of treating cancer in a patient, wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma, comprising administering to the patient in need of such a treatment, a combination of necitumumab and abemaciclib, or a pharmaceutically acceptable salt thereof, wherein necitumumab is administered at a dose of 800 mg, and wherein abemaciclib is administered at a dose of 50 mg to 200 mg.

3. The method of claim 2, wherein necitumumab is administered at a dose of 800 mg on days 1 and 8 of each 3-week cycle by intravenous infusion.

4. The method of claim 2, wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered twice a day.

5. The method of claim 2, wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is administered at a dose of 50 mg.

6. The method of claim 2, wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is administered at a dose of 100 mg.

7. The method of claim 2, wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is administered at a dose of 150 mg.

8. The method of claim 2, wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is administered at a dose of 200 mg.

9. The method of claim 1 or 2, wherein the combination of necitumumab and

abemaciclib, or a pharmaceutically acceptable salt thereof, is administered simultaneously, separately, or sequentially.

10. Necitumumab for use in simultaneous, separate, or sequential combination with abemaciclib, or a pharmaceutically acceptable salt thereof, in the treatment of cancer, wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma.

11. Necitumumab for the use of claim 10, wherein necitumumab is administered at a dose of 800 mg by intravenous infusion.

12. Necitumumab for the use of claims 10 or 11, wherein abemaciclib, or a

pharmaceutically acceptable salt thereof, is orally administered at a dose of 50 mg to 200 mg.

13. Necitumumab for the use of any one of claims 10 to 12, wherein necitumumab is administered at a dose of 800 mg by intravenous infusion and wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered at a dose of 50 mg to 200 mg.

14. Necitumumab for the use of any one of claims 10 to 13, wherein necitumumab is administered at a dose of 800 mg on days 1 and 8 of each 3 -week cycle by intravenous infusion and wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered at a dose of 50 mg to 200 mg twice a day.

15. Abemaciclib, or a pharmaceutically acceptable salt thereof, for use in

simultaneous, separate, or sequential combination with necitumumab for the treatment of cancer, wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered at a dose of 50 mg to 200 mg and wherein necitumumab is administered at a dose of 800 mg by intravenous infusion, and wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma.

16. Abemaciclib, or a pharmaceutically acceptable salt thereof, for the use of claim 15, wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered at a dose of 50 mg to 200 mg twice a day and wherein necitumumab is administered at a dose of 800 mg by intravenous infusion on days 1 and 8 of each 3-week cycle.

17. Abemaciclib, or a pharmaceutically acceptable salt thereof, for the use of claim 15 or 16, wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is administered at a dose of 50 mg.

18. Abemaciclib, or a pharmaceutically acceptable salt thereof, for the use of claim 15 or 16, wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is administered at a dose of 100 mg.

19. Abemaciclib, or a pharmaceutically acceptable salt thereof, for the use of claim 15 or 16, wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is administered at a dose of 150 mg.

20. Abemaciclib, or a pharmaceutically acceptable salt thereof, for the use of claim 15 or 16, wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is administered at a dose of 200 mg.

21. A kit comprising necitumumab and abemaciclib, or a pharmaceutically acceptable salt thereof, for use in the treatment of cancer, wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma.

22. The kit for the use of claim 21, wherein the amount of necitumumab is 800 mg, and the amount of abemaciclib, or a pharmaceutically acceptable salt thereof, is 50 mg to 200 mg.

23. The kit for the use of claim 21, wherein the amount of abemaciclib, or a

pharmaceutically acceptable salt thereof, is 50 mg.

24. The kit for the use of claim 21, wherein the amount of abemaciclib, or a

pharmaceutically acceptable salt thereof, is 100 mg.

25. The kit for the use of claim 21, wherein the amount of abemaciclib, or a

pharmaceutically acceptable salt thereof, is 150 mg.

26. The kit for the use of claim 21, wherein the amount of abemaciclib, or a

pharmaceutically acceptable salt thereof, is 200 mg.

27. A kit, comprising a pharmaceutical composition, comprising necitumumab with one or more pharmaceutically acceptable carriers, diluents, or excipients, and a pharmaceutical composition, comprising abemaciclib, or a pharmaceutically acceptable salt thereof, with one or more pharmaceutically acceptable carriers, diluents, or excipients for use in the treatment of cancer, wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma.

28. The kit for the use of claim 27, wherein the amount of necitumumab is 800 mg, and wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is 50 mg to 200 mg.

29. The kit for the use of claim 28, wherein the amount of abemaciclib, or a

pharmaceutically acceptable salt thereof, is 50 mg.

30. The kit for the use of claim 28, wherein the amount of abemaciclib, or a

pharmaceutically acceptable salt thereof, is 100 mg.

31. The kit for the use of claim 28, wherein the amount of abemaciclib, or a

pharmaceutically acceptable salt thereof, is 150 mg.

32. The kit for the use of claim 28, wherein the amount of abemaciclib, or a

pharmaceutically acceptable salt thereof, is 200 mg.

33. A kit comprising necitumumab and abemaciclib, or a pharmaceutically acceptable salt thereof, wherein necitumumab is formulated at a final concentration of 16 mg/mL in a formulation of 10 mM citrate, 40 mM sodium chloride, 133 mM glycine, 50 mM mannitol, 0.01% polysorbate-80, pH6.0; and wherein

abemaciclib, or a pharmaceutically acceptable salt thereof, is supplied as capsules or tablets of 50 mg of abemaciclib per capsule or tablet.

34. The method of claims 1-9, necitumumab for use of claims 10-14, abemaciclib for use of claims 15-20, the kit for the use of claims 21-26, and the kit for the use of claims 27-32, wherein the non-small cell lung cancer is squamous.

35. The method of claims 1-9, necitumumab for use of claims 10-14, abemaciclib for use of claims 15-20, the kit for the use of claims 21-26, and the kit for the use of claims 27-32, wherein the non-small cell lung cancer is non-squamous.

36. The method of claims 1-9, necitumumab for use of claims 10-14, abemaciclib for use of claims 15-20, the kit for the use of claims 21-26, and the kit for the use of claims 27-32, wherein the head and neck cancer is squamous.

37. A method of treating cancer in a patient, wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma, comprising administering to the patient in need of such a treatment, a combination of necitumumab and abemaciclib, or a pharmaceutically acceptable salt thereof, wherein necitumumab is administered at a dose of 800 mg, and wherein abemaciclib is administered at a dose of 150 mg.

38. The method of claim 37, wherein necitumumab is administered on days 1 and 8 of a 3-week cycle by intravenous infusion.

39. The method of claim 37, wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered twice a day.

40. The method of claim 37, wherein the combination of necitumumab and

41. Necitumumab for use in simultaneous, separate, or sequential combination with abemaciclib, or a pharmaceutically acceptable salt thereof, in the treatment of cancer, wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma, wherein necitumumab is administered at a dose of 800 mg, and wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is administered at a dose of 150 mg.

42. Necitumumab for the use of claim 41, wherein necitumumab is administered on days 1 and 8 of a 3-week cycle by intravenous infusion, and wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered twice a day.

43. Abemaciclib, or a pharmaceutically acceptable salt thereof, for use in

simultaneous, separate, or sequential combination with necitumumab for the treatment of cancer, wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is administered at a dose of 150 mg, wherein necitumumab is

administered at a dose of 800 mg, and wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma.

44. Abemaciclib, or a pharmaceutically acceptable salt thereof, for the use of claim 43, wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is orally administered twice a day, and wherein necitumumab is administered by intravenous infusion on days 1 and 8 of a 3 -week cycle.

45. A kit comprising necitumumab and abemaciclib, or a pharmaceutically acceptable salt thereof, for use in the treatment of cancer, wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma, and wherein the amount of necitumumab is 800 mg, and the amount of abemaciclib, or a pharmaceutically acceptable salt thereof, is 50, 100, or 150 mg.

46. A kit, comprising a pharmaceutical composition, comprising necitumumab with one or more pharmaceutically acceptable carriers, diluents, or excipients, and a pharmaceutical composition, comprising abemaciclib, or a pharmaceutically acceptable salt thereof, with one or more pharmaceutically acceptable carriers, diluents, or excipients for use in the treatment of cancer, wherein the cancer is selected from the group consisting of non-small cell lung cancer, colorectal cancer, head and neck cancer, and pancreatic carcinoma, wherein the amount of necitumumab is 800 mg, and wherein abemaciclib, or a pharmaceutically acceptable salt thereof, is 50, 100, or 150 mg.

47. A kit comprising necitumumab and abemaciclib, or a pharmaceutically acceptable salt thereof, wherein necitumumab is formulated at a final concentration of 16 mg/mL in a formulation of 10 mM citrate, 40 mM sodium chloride, 133 mM glycine, 50 mM mannitol, 0.01% polysorbate-80, pH6.0; and wherein

abemaciclib, or a pharmaceutically acceptable salt thereof, is supplied as capsules or tablets of 50 mg, 100 mg, or 150 mg of abemaciclib per capsule or tablet.

48. The method of claims 37-40, necitumumab for use of claims 41-42, abemaciclib for use of claims 43-44, the kit for the use of claim 45, the kit for the use of claim 46, and the kit for the use of claim 47, wherein the non-small cell lung cancer is squamous.

49. The method of claims 37-40, necitumumab for use of claims 41-42, abemaciclib for use of claims 43-44, the kit for the use of claim 45, the kit for the use of claim 46, and the kit for the use of claim 47, wherein the non-small cell lung cancer is non-squamous.

50. The method of claims 37-40, necitumumab for use of claims 41-42, abemaciclib for use of claims 43-44, the kit for the use of claim 45, the kit for the use of claim 46, and the kit for the use of claim 47, wherein the head and neck cancer is squamous.