WO2017205213A1

WO2017205213A1 - Combination therapy of abemaciclib and immune checkpoint modulators for use in the treatment of cancer

Info

Publication number: WO2017205213A1
Application number: PCT/US2017/033594
Authority: WO
Inventors: Richard Paul Beckmann; David Arlen SCHAER
Original assignee: Eli Lilly And Company
Priority date: 2016-05-23
Filing date: 2017-05-19
Publication date: 2017-11-30

Abstract

The present invention relates to a combination of LY3300054 and abemaciclib, or a pharmaceutically acceptable salt thereof, and to methods of using these combinations to treat certain disorders, in particular any cancer that is sensitive to CDK4/6 inhibitors for example, non-small cell lung cancer (NSCLC), breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkins lymphoma such as mantle cell lymphoma, and tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient.

Description

COMBINATION THERAPY OF ABEMACICLIB AND IMMUNE CHECKPOINT MODULATORS FOR USE IN THE TREATMENT OF CANCER

The present invention relates to a combination of an anti-Programmed cell Death 1 Ligand 1 (PD-L1) antibody, LY3300054, and abemaciclib and to methods of using the combination to treat certain disorders, in particular any cancer that is sensitive to CDK4/6 inhibitors for example, non-small cell lung cancer (NSCLC), breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6- Rb-E2F pathway), and additional tumors which display immune inflammation after a treatment with abemaciclib in a patient.

The combination described herein may provide a beneficial effect, for example, in the treatment of a cancer, such as an enhanced anti-cancer effect, reduced toxicity and/or reduced adverse effects. For example, LY3300054, abemaciclib, or a pharmaceutically acceptable salt thereof, or all of the above, may be administered at a lower dosage, or for reduced duration than would be required to achieve the same therapeutic effect compared to a monotherapy dose. In addition, combination treatments as compared to monotherapy may lead to higher overall response rates, increased progression-free survival and/or increased overall survival.

According to an aspect of the present invention, there is presented a method of treating a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6- Rb-E2F pathway), and additional susceptible tumors in a patient, comprising

administering to the patient an effective amount of LY3300054 and the abemaciclib or the salt thereof, wherein initial administration of the abemaciclib or the salt thereof is administered to the patient without the LY3300054 for at least 24 hours and the abemaciclib or the salt thereof is then administered to the patient in combination with the LY3300054. Preferably the abemaciclib or the salt thereof is administered orally at 150 mg approximately every 12 hours. Preferably the abemaciclib or the salt thereof is administered orally at 100 mg approximately every 12 hours. Preferably the LY3300054 is administered at 70 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 700 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1000 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1400 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at a dose level needed to achieve constant saturation of available PD-L1 target protein by intravenous infusion during the course of combination therapy. Preferably the abemaciclib or the salt thereof is administered alone for 2 weeks of a first 28-day cycle and then starting on Day 15 of the first 28-day cycle and thereafter on Days 1 and 15 of subsequent cycles, the abemaciclib or the salt thereof is administered in combination with the LY3300054. Preferably on days the abemaciclib or the salt thereof and the LY3300054 are administered on the same day, the abemaciclib or the salt thereof is administered approximately 2 hours after starting the administration of the LY3300054.

The following table illustrates additional preferences for the above method. These preferences may be combined with one another as further preferences as illustrated for instance in the paragraph after this table. Table 1

A: Compound B: Cancer C: Antibody D: Initial administration

of abemaciclib, or the

salt thereof

1 Abemaciclib, or NSCLC LY3300054 Initial administration for at

a least 1-28 days

pharmaceutically

acceptable salt

thereof

2 breast Initial administration for a

time necessary to induce

stable disease or partial

response as determined by

RECIST criteria or as

determined by treating

physician

3 pancreatic Initial administration for a

period of time which

disease progression occurs

as measured by RECIST

criteria or as determined

by treating physician 4 mantle cell Initial administration for a

lymphoma time necessary to induce

intra-tumor immune

inflammation

5 tumors which Initial administration for a

display time necessary to increase immune expression of immune inflammation inflammatory genes

after a

treatment with

abemaciclib

or a salt

thereof

6 tumors which Initial administration of

display the abemaciclib or the salt alterations in thereof to the patient is for cell cycle a period of time necessary,

control prior to initiation of the

(CyclinD- combination, that the

CDK4/CDK6- subsequent combination

Rb-E2F therapy of the abemaciclib pathway) or the salt thereof with

LY3300054 induces intra- tumor immune

inflammation.

Further preferred: Al, Bl, CI, and Dl; Al, B2, CI, and Dl; Al, B3, C2, and Dl; Al, B4, CI, and Dl; Al, B5, CI, and Dl; Al, B6, CI, and Dl; Al, Bl, CI, and D2; Al, B2, CI, and D2; Al, B3, CI, and D2; Al, B4, CI, and D2; Al, B5, CI, and D2; Al, B6, CI, and D2; Al, Bl, CI, and D3; Al, B2, CI, and D3; Al, B3, CI, and D3; Al, B4, CI, and D3; Al, B5, CI, and D3; Al, B6, CI, and D3; Al, Bl, CI, and D4; Al, B2, CI, and D4; Al, B3, CI, and D4; Al, B4, CI, and D4; Al, B5, CI, and D4; Al, B6, CI, and D4; Al, Bl, CI, and D5; Al, B2, CI, and D5; Al, B3, CI, and D5; Al, B4, CI, and D5; Al, B5, CI, and D5; Al, B6, CI, and D5; Al, Bl, CI, and D6; Al, B2, CI, and D6; Al, B3, CI, and D6; Al, B4, CI, and D6; Al, B5, CI, and D6; and Al, B6, CI, and D6;

In the above aspects and preferences, optionally, the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof and the LY3300054.

In the above aspects and preferences, optionally, the administration of the abemaciclib or the salt thereof is stopped after initiation of the combination with the LY3300054 without administering the abemaciclib or the salt thereof. According to another aspect of the present invention, there is presented a kit comprising LY3300054 and abemaciclib, or a pharmaceutically acceptable salt thereof for the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin' s lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD- CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient.

According to another aspect of the present invention, there is presented a kit comprising LY3300054 and abemaciclib, or a pharmaceutically acceptable salt thereof, for the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin' s lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD- CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient.

For the aforementioned kits, preferably, the kit comprises abemaciclib, or a pharmaceutically acceptable salt thereof, and LY3300054. Preferably, the kit comprises the abemaciclib or the salt thereof and the LY3300054 for the treatment of NSCLC.

Preferably, the kit comprises the abemaciclib or the salt thereof and the LY3300054 for the treatment of breast cancer. Preferably, the kit comprises the abemaciclib or the salt thereof, and the LY3300054 for the treatment of pancreatic cancer. Preferably, the kit comprises the abemaciclib or the salt thereof and the LY3300054 for the treatment of mantle cell lymphoma. Preferably, the kit comprises the abemaciclib or the salt thereof and the LY3300054 for the treatment of tumors which display immune inflammation after a treatment with the abemaciclib or the salt thereof. Preferably, the kit comprises the abemaciclib or the salt thereof and the LY3300054 for the treatment of tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway).

Preferably, the kit comprises the abemaciclib or the salt thereof and the LY3300054 for the treatment of additional susceptible tumors in a patient.

According to another aspect of the present invention, there is presented a combination of LY3300054 and abemaciclib, or a pharmaceutically acceptable salt thereof, for simultaneous, separate, or sequential use in the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin' s lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors wherein initial administration of the abemaciclib or the salt thereof is administered to the patient without the LY3300054 for at least 24 hours and the abemaciclib or the salt thereof is then administered to the patient in simultaneous, separate, or sequential combination with the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof, and/or the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped after initiation of the combination with the LY3300054, and administration of the LY3300054 is continued without administering the abemaciclib or the salt thereof.

Preferably the abemaciclib or the salt thereof is administered orally at 150 mg approximately every 12 hours. Preferably the abemacicilib or the salt thereof is administered orally at 100 mg approximately every 12 hours. Preferably the LY3300054 is administered at 70 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 700 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1000 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1400 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at a dose level needed to achieve constant saturation of available PD-L1 target protein by intravenous infusion during the course of combination therapy. Preferably the abemaciclib or the salt thereof is administered alone for 2 weeks of a first 28-day cycle and then starting on Day 15 of the first 28-day cycle and thereafter on Days 1 and 15 of subsequent cycles, the abemaciclib or the salt thereof is administered in combination with the LY3300054. Preferably on days the abemaciclib or the salt thereof and the LY3300054 are administered on the same day, the abemaciclib or the salt thereof is administered approximately 2 hours after starting the administration of the LY3300054.

According to another aspect of the present invention, there is presented a combination comprising LY3300054 and abemaciclib, or a pharmaceutically acceptable salt thereof, for simultaneous, separate, or sequential use in the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein initial administration of the abemaciclib or the salt thereof is administered to the patient without the LY3300054 for at least 1 to 28 days and the abemaciclib or the salt thereof is then administered to the patient in simultaneous, separate, or sequential combination with the LY3300054.

Optionally, the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof and/or the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped after initiation of the combination with the LY3300054, and administration of the LY3300054 is continued without administering the abemaciclib or the salt thereof. Preferably the abemaciclib or the salt thereof is administered orally at 150 mg approximately every 12 hours. Preferably the abemaciclib or the salt thereof is administered orally at 100 mg approximately every 12 hours.

Preferably the LY3300054 is administered at 70 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 700 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1000 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1400 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at a dose level needed to achieve constant saturation of available PD-L1 target protein by intravenous infusion during the course of combination therapy. Preferably the abemaciclib or the salt thereof is administered alone for 2 weeks of a first 28-day cycle and then starting on Day 15 of the first 28-day cycle and thereafter on Days 1 and 15 of subsequent cycles, the abemaciclib or the salt thereof is administered in combination with the LY3300054. Preferably on days the abemaciclib or the salt thereof and the LY3300054 are administered on the same day, the abemaciclib or the salt thereof is administered approximately 2 hours after starting the administration of the LY3300054.

According to another aspect of the present invention, there is presented a combination comprising LY3300054 and abemaciclib, or a pharmaceutically acceptable salt thereof, for simultaneous, separate, or sequential use in the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein initial administration of the abemaciclib or the salt thereof is administered to the patient without the LY3300054 for a time necessary to induce stable disease or partial response as determined by RECIST criteria or as determined by treating physician and the abemaciclib or the salt thereof is then administered to the patient in simultaneous, separate, or sequential combination with the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof and/or the LY3300054. Optionally, the

administration of the abemaciclib or the salt thereof is stopped after initiation of the combination with the LY3300054, and administration of the LY3300054 is continued without administering the abemaciclib or the salt thereof. Preferably the abemaciclib or the salt thereof is administered orally at 150 mg approximately every 12 hours.

Preferably the abemaciclib or the sat thereof is administered orally at 100 mg

approximately every 12 hours. Preferably the LY3300054 is administered at 70 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 700 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1000 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1400 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at a dose level needed to achieve constant saturation of available PD-L1 target protein by intravenous infusion during the course of combination therapy. Preferably the abemaciclib or the salt thereof is administered alone for 2 weeks of a first 28-day cycle and then starting on Day 15 of the first 28-day cycle and thereafter on Days 1 and 15 of subsequent cycles, the abemaciclib or the salt thereof is administered in combination with the LY3300054. Preferably on days the abemaciclib or the salt thereof and the LY3300054 are administered on the same day, the abemaciclib or the salt thereof is administered approximately 2 hours after starting the administration of the LY3300054.

According to another aspect of the present invention, there is presented a combination comprising LY3300054 and abemaciclib, or a pharmaceutically acceptable salt thereof, for simultaneous, separate, or sequential use in the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein initial administration of the abemaciclib or the salt thereof is administered to the patient without the LY3300054 for a period of time which disease progression occurs as measured by RECIST criteria or as determined by treating physician and the abemaciclib or the salt thereof is then administered to the patient in simultaneous, separate, or sequential combination with the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof and/or the LY3300054. Optionally, the

administration of the abemaciclib or the salt thereof is stopped after initiation of the combination with the LY3300054 and administration of the LY3300054 is continued without administering the abemaciclib or the salt thereof. Preferably the abemaciclib or the salt thereof is administered orally at 150 mg approximately every 12 hours.

Preferably the abemaciclib or the salt thereof is administered orally at 100 mg approximately every 12 hours. Preferably the LY3300054 is administered at 70 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 700 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1000 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1400 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at a dose level needed to achieve constant saturation of available PD-L1 target protein by intravenous infusion during the course of combination therapy. Preferably the abemaciclib or the salt thereof is administered alone for 2 weeks of a first 28-day cycle and then starting on Day 15 of the first 28-day cycle and thereafter on Days 1 and 15 of subsequent cycles, the abemaciclib or the salt thereof is administered in combination with the LY3300054. Preferably on days the abemaciclib or the salt thereof and the

LY3300054 are administered on the same day, the abemaciclib or the salt thereof is administered approximately 2 hours after starting the administration of the LY3300054.

According to another aspect of the present invention, there is presented a combination comprising LY3300054 and abemaciclib, or a pharmaceutically acceptable salt thereof, for simultaneous, separate, or sequential use in the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein initial administration of the abemaciclib or the salt thereof, is administered to the patient without the LY3300054 for a time necessary to induce intra-tumor immune inflammation and the abemaciclib or the salt thereof, is then administered to the patient in simultaneous, separate, or sequential combination with the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof and/or the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped after initiation of the combination with the LY3300054, and administration of the LY3300054 is continued without administering the abemaciclib or the salt thereof. Preferably the abemaciclib or the salt thereof is administered orally at 150 mg approximately every 12 hours. Preferably the abemaciclib or the salt thereof is administered orally at 100 mg approximately every 12 hours. Preferably the LY3300054 is administered at 70 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 700 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1000 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1400 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at a dose level needed to achieve constant saturation of available PD-L1 target protein by intravenous infusion during the course of combination therapy. Preferably the abemaciclib or the salt thereof is administered alone for 2 weeks of a first 28-day cycle and then starting on Day 15 of the first 28-day cycle and thereafter on Days 1 and 15 of subsequent cycles, the abemaciclib or the salt thereof is administered in combination with the LY3300054. Preferably on days the abemaciclib or the salt thereof and the

According to another aspect of the present invention, there is presented a combination comprising LY3300054 and abemaciclib, or a pharmaceutically acceptable salt thereof, for simultaneous, separate, or sequential use in the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein initial administration of the abemaciclib or the salt thereof is administered to the patient without the LY3300054 for a time necessary to increase expression of immune inflammatory genes and the abemaciclib or the salt thereof is then administered to the patient in simultaneous, separate, or sequential combination with the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the compound or the salt thereo, and/or the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped after initiation of the combination with the LY3300054, and administration of the LY3300054 is continued without administering the abemaciclib or the salt thereof.

Preferably the abemaciclib or the salt thereof is administered orally at 150 mg approximately every 12 hours. Preferaby the abemaciclib or the salt thereof is administered orally at 100 mg approximately every 12 hours. Preferably the LY3300054 is administered at 70 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 700 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1000 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1400 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at a dose level needed to achieve constant saturation of available PD-L1 target protein by intravenous infusion during the course of combination therapy. Preferably the abemaciclib or the salt thereof is administered alone for 2 weeks of a first 28-day cycle and then starting on Day 15 of the first 28-day cycle and thereafter on Days 1 and 15 of subsequent cycles, the abemaciclib or the salt thereof is administered in combination with the LY3300054. Preferably on days the abemaciclib or the salt thereof and the LY3300054 are administered on the same day, the abemaciclib or the salt thereof is administered approximately 2 hours after starting the administration of the LY3300054.

According to another aspect of the present invention, there is presented a combination comprising LY3300054 and abemaciclib, or a pharmaceutically acceptable salt thereof, for simultaneous, separate, or sequential use in the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein initial administration of the abemaciclib or the salt thereof is administered to the patient without the LY3300054 for a period of time necessary, prior to initiation of the combination, that the subsequent combination therapy of the abemaciclib or the salt thereof with the LY3300054 induces intra-tumor immune inflammation and the abemaciclib or the salt thereof, is then administered to the patient in simultaneous, separate, or sequential combination with the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof, is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof and/or the LY3300054. Optionally, the

According to another aspect of the present invention, there is presented an anti-

PD-1 antibody which is LY3300054 for use in simultaneous, separate, or sequential combination with abemaciclib, or a pharmaceutically acceptable salt thereof, in the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin' s lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-

CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein the initial administration of the abemaciclib or the salt thereof is administered to the patient without the LY3300054 for at least 24 hours and the abemaciclib or the salt thereof, is then administered to the patient in simultaneous, separate, or sequential combination with the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof and/or the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped after initiation of the combination with the LY3300054, and administration of the LY3300054 is continued without administering the abemaciclib or the salt thereof. Preferably the abemaciclib or the salt thereof is administered orally at 150 mg approximately every 12 hours. Preferably the abemaciclib or the salt thereof is administered orally at 100 mg approximately every 12 hours. Preferably the LY3300054 is administered at 70 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 700 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1000 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1400 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at a dose level needed to achieve constant saturation of available PD-L1 target protein by intravenous infusion during the course of combination therapy. Preferably the abemaciclib or the salt thereof is administered alone for 2 weeks of a first 28-day cycle and then starting on Day 15 of the first 28-day cycle and thereafter on Days 1 and 15 of subsequent cycles, the abemaciclib or the salt thereof is administered in combination with the LY3300054. Preferably on days the abemaciclib or the salt thereof and the

According to another aspect of the present invention, there is presented

LY3300054 for use in simultaneous, separate, or sequential combination with abemaciclib, or a pharmaceutically acceptable salt thereof, in the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein the initial administration of the abemaciclib or the salt thereof is administered to the patient without the LY3300054 for at least 1 to 28 days and the abemaciclib or the salt thereof is then administered to the patient in simultaneous, separate, or sequential combination with the LY3300054.

According to another aspect of the present invention, there is LY3300054 for use in simultaneous, separate, or sequential combination with abemaciclib, or a

pharmaceutically acceptable salt thereof, in the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin' s lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein the initial administration of the abemaciclib or the salt thereof is administered to the patient without the LY3300054 for a time necessary to induce stable disease or partial response as determined by RECIST criteria or as determined by treating physician and the abemaciclib or the salt thereof is then administered to the patient in simultaneous, separate, or sequential combination with the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof and/or the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof, is stopped after initiation of the combination with the LY3300054, and administration of the LY3300054 is continued without administering the abemaciclib or the salt thereof. Preferably the abemaciclib or the salt thereof is administered orally at 150 mg approximately every 12 hours.

According to another aspect of the present invention, there is presented

LY3300054 for use in simultaneous, separate, or sequential combination with abemaciclib, or a pharmaceutically acceptable salt thereof, in the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein the initial administration of the abemaciclib or the salt thereof is administered to the patient without the LY3300054 for a period of time which disease progression occurs as measured by RECIST criteria or as determined by treating physician and the abemaciclib or the salt thereof is then administered to the patient in simultaneous, separate, or sequential combination with the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof and/or the LY3300054. Optionally, the

According to another aspect of the present invention, there is presented

LY3300054 for use in simultaneous, separate, or sequential combination with

abemaciclib, or a pharmaceutically acceptable salt thereof, in the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein the initial administration of the abemaciclib or the salt thereof is administered to the patient without the LY3300054 for a time necessary to induce intra-tumor immune inflammation and the abemaciclib or the salt thereof is then administered to the patient in simultaneous, separate, or sequential combination with the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof, and/or the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped after initiation of the combination with the LY3300054, and administration of the LY3300054 is continued without administering the abemaciclib or the salt thereof. Preferably the abemaciclib or the salt thereof is administered orally at 150 mg approximately every 12 hours. Preferably the abemaciclib or the salt thereof is administered orally at 100 mg approximately every 12 hours. Preferably the LY3300054 is administered at 70 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 700 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1000 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1400 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at a dose level needed to achieve constant saturation of available PD-L1 target protein by intravenous infusion during the course of combination therapy. Preferably the abemaciclib or the salt thereof is administered alone for 2 weeks of a first 28-day cycle and then starting on Day 15 of the first 28-day cycle and thereafter on Days 1 and 15 of subsequent cycles, the abemaciclib or the salt thereof is administered in combination with the LY3300054. Preferably on days the abemaciclib or the salt thereof and the

According to another aspect of the present invention, there is presented

LY3300054 for use in simultaneous, separate, or sequential combination with abemaciclib, or a pharmaceutically acceptable salt thereof, in the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein the initial administration of the abemaciclib or the salt thereof is administered to the patient without the LY3300054 for a time necessary to increase expression of immune inflammatory genes and the abemaciclib or the salt thereof, is then administered to the patient in simultaneous, separate, or sequential combination with the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof and/or the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped after initiation of the combination with the LY3300054, and administration of the LY3300054 is continued without administering the abemaciclib or the salt thereof.

Preferably the abemaciclib or the salt thereof is administered orally at 150 mg approximately every 12 hours. Preferably the abemaciclib or the salt thereof is administered orally at 100 mg approximately every 12 hours. Preferably the LY3300054 is administered at 70 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 700 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1000 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1400 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at a dose level needed to achieve constant saturation of available PD-L1 target protein by intravenous infusion during the course of combination therapy. Preferably the abemaciclib or the salt thereof is administered alone for 2 weeks of a first 28-day cycle and then starting on Day 15 of the first 28-day cycle and thereafter on Days 1 and 15 of subsequent cycles, the abemaciclib or the salt thereof is administered in combination with the LY3300054. Preferably on days the abemaciclib or the salt thereof and the LY3300054 are administered on the same day, the abemaciclib or the salt thereof is administered approximately 2 hours after starting the administration of the LY3300054..

According to another aspect of the present invention, there is presented

LY3300054 for use in simultaneous, separate, or sequential combination with abemaciclib, or a pharmaceutically acceptable salt thereof, in the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein the initial administration of the abemaciclib or the salt thereof, is administered to the patient without the LY3300054 for a period of time necessary, prior to initiation of the combination, that the subsequent combination therapy of the abemaciclib or the salt thereof with the LY3300054 induces intra-tumor immune inflammation and the abemaciclib or the salt thereof is then administered to the patient in simultaneous, separate, or sequential combination with the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof and/or the LY3300054. Optionally, the

Preferably the abemaciclib or the salt thereof is administered orally at 100 mg approximately every 12 hours. Preferably the LY3300054 is administered at 70 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 700 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1000 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1400 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at a dose level needed to achieve constant saturation of available PD-L1 target protein by intravenous infusion during the course of combination therapy. Preferably the abemaciclib or the salt thereof is administered alone for 2 weeks of a first 28-day cycle and then starting on Day 15 of the first 28-day cycle and thereafter on Days 1 and 15 of subsequent cycles, the abemaciclib or the salt thereof is administered in combination with the LY3300054. Preferably on days the abemaciclib or the salt thereof and the LY3300054 are administered on the same day, the abemaciclib or the salt thereof is administered approximately 2 hours after starting the administration of the LY3300054.

According to another aspect of the present invention, there is presented abemaciclib, or a pharmaceutically acceptable salt thereof, for use in simultaneous, separate, or sequential combination with LY3300054 in the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein the initial administration of the abemaciclib or the salt thereof is administered without the LY3300054 for at least 24 hours and the abemaciclib or the salt thereof is then administered in simultaneous, separate, or sequential combination with the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof, is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof, and/or the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped after initiation of the combination with the LY3300054, and administration of the LY3300054 is continued without administering the abemaciclib or the salt thereof.

Preferably the abemaciclib or the salt thereof is administered orally at 150 mg approximately every 12 hours. Preferably the abemaciclib or the salt thereof is administered orally at 100 mg approximately every 12 hours. Preferably the LY3300054 is administered at 70 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 700 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1000 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1400 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at a dose level needed to achieve constant saturation of available PD-L1 target protein by intravenous infusion during the course of combination therapy. Preferably the abemaciclib or the salt thereof is administered alone for 2 weeks of a first 28-day cycle and then starting on Day 15 of the first 28-day cycle and thereafter on Days 1 and 15 of subsequent cycles, the abemaciclib or the salt thereof is administered in combination with the LY3300054. Preferably on days the abemaciclib or the salt thereof and the LY3300054 are administered on the same day, the abemaciclib or the salt thereof is administered approximately 2 hours after starting the administration of the LY3300054.

According to another aspect of the present invention, there is presented abemaciclib, or a pharmaceutically acceptable salt thereof, for use in simultaneous, separate, or sequential combination with LY3300054 in the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein the initial administration of the abemaciclib or the salt thereof is administered without the LY3300054 for at least 1 to 28 days and the abemaciclib or the salt thereof is then administered in simultaneous, separate, or sequential combination with the LY3300054. Optionally, the administration of the compouund or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof and/or the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped after initiation of the combination with the LY3300054, and administration of the LY3300054 is continued without administering the abemaciclib or the salt thereof.

According to another aspect of the present invention, there is presented abemaciclib, or a pharmaceutically acceptable salt thereof, for use in simultaneous, separate, or sequential combination with LY3300054 in the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein the initial administration of the abemaciclib or the salt thereof, is administered without the LY3300054 for a time necessary to induce stable disease or partial response as determined by RECIST criteria or as determined by treating physician and the abemaciclib or the salt thereof is then administered in simultaneous, separate, or sequential combination with the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof and/or the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof, is stopped after initiation of the combination with the LY3300054, and administration of the LY3300054 is continued without administering the abemaciclib or the salt thereof.

According to another aspect of the present invention, there is presented abemaciclib, or a pharmaceutically acceptable salt thereof, for use in simultaneous, separate, or sequential combination with LY3300054 in the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein the initial administration of the abemaciclib or the salt thereof is administered without the LY3300054 for a period of time which disease progression occurs as measured by RECIST criteria or as determined by treating physician and the abemaciclib or the salt thereof, is then administered in simultaneous, separate, or sequential combination with the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof and/or the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped after initiation of the combination with the LY3300054, and administration of the LY3300054 is continued without administering the abemaciclib or the salt thereof.

According to another aspect of the present invention, there is presented abemaciclib, or a pharmaceutically acceptable salt thereof, for use in simultaneous, separate, or sequential combination with LY3300054 in the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein the initial administration of the abemaciclib or the salt thereof is administered without the LY3300054 for a time necessary to induce intra- tumor immune inflammation and the abemaciclib or the salt thereof is then administered in simultaneous, separate, or sequential combination with the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof, and/or the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped after initiation of the combination with the LY3300054, and administration of the LY3300054 is continued without administering the abemaciclib or the salt thereof. Preferably the abemaciclib or the salt thereof is administered orally at 150 mg approximately every 12 hours. Preferably the abemaciclib or the salt thereof is administered orally at 100 mg approximately every 12 hours. Preferably the LY3300054 is administered at 70 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 700 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1000 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1400 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at a dose level needed to achieve constant saturation of available PD-L1 target protein by intravenous infusion during the course of combination therapy. Preferably the abemaciclib or the salt thereof is administered alone for 2 weeks of a first 28-day cycle and then starting on Day 15 of the first 28-day cycle and thereafter on Days 1 and 15 of subsequent cycles, the abemaciclib or the salt thereof is administered in combination with the LY3300054. Preferably on days the abemaciclib or the salt thereof and the LY3300054 are administered on the same day, the abemaciclib or the salt thereof is administered approximately 2 hours after starting the administration of the LY3300054.

According to another aspect of the present invention, there is presented abemaciclib, or a pharmaceutically acceptable salt thereof, for use in simultaneous, separate, or sequential combination with LY3300054 in the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein the initial administration of the abemaciclib or the salt thereof is administered without the LY3300054 for a time necessary to increase expression of immune inflammatory genes and the abemaciclib or the salt thereof is then administered in simultaneous, separate, or sequential combination with the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof, and/or the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped after initiation of the combination with the LY3300054, and administration of the LY3300054 is continued without administering the abemaciclib or the salt thereof. Preferably the abemaciclib or the salt thereof is administered orally at 150 mg approximately every 12 hours. Preferably the abemaciclib or the salt thereof is administered orally at 100 mg approximately every 12 hours. Preferably the LY3300054 is administered at 70 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 700 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1000 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1400 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at a dose level needed to achieve constant saturation of available PD-L1 target protein by intravenous infusion during the course of combination therapy. Preferably the abemaciclib or the salt thereof is administered alone for 2 weeks of a first 28-day cycle and then starting on Day 15 of the first 28-day cycle and thereafter on Days 1 and 15 of subsequent cycles, the abemaciclib or the salt thereof is administered in combination with the LY3300054. Preferably on days the abemaciclib or the salt thereof and the LY3300054 are administered on the same day, the abemaciclib or the salt thereof is administered approximately 2 hours after starting the administration of the LY3300054.

According to another aspect of the present invention, there is presented abemaciclib, or a pharmaceutically acceptable salt thereof, for use in simultaneous, separate, or sequential combination with LY3300054 in the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein the initial administration of the abemaciclib or the salt thereof is administered without the LY3300054 for a period of time necessary, prior to initiation of the combination, that the subsequent combination therapy of the abemaciclib or the salt thereof with the LY3300054 induces intra-tumor immune inflammation and the abemaciclib or the salt thereof is then administered in simultaneous, separate, or sequential combination with the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof, and/or the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped after initiation of the combination with the LY3300054, and administration of the LY3300054 is continued without administering the abemaciclib or the salt thereof.

According to another aspect of the present invention, there is presented the use of LY3300054 in the manufacture of a medicament for the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein the LY3300054 is administered in simultaneous, separate, or sequential combination with abemaciclib, or a pharmaceutically acceptable salt thereof, after an initial administration of the abemaciclib or the salt thereof is administered without the LY3300054 for at least 24 hours. Optionally, the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof and/or the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped after initiation of the combination with the LY3300054, and administration of the LY3300054 is continued without administering the abemaciclib or the salt thereof.

According to another aspect of the present invention, there is presented the use of LY3300054 in the manufacture of a medicament for the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein the LY3300054 is administered in simultaneous, separate, or sequential combination with abemaciclb, or a pharmaceutically acceptable salt thereof, after an initial administration of the abemaciclib or the salt thereof is administered without the LY3300054 for at least 1 to 28 days. Optionally, the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof and/or the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped after initiation of the combination with the LY3300054, and administration of the LY3300054 is continued without administering the abemaciclib or the salt thereof. Preferably the abemaciclib or the salt thereof is administered orally at 150 mg approximately every 12 hours. Preferably the abemaciclib or the salt thereof is administered orally at 100 mg approximately every 12 hours. Preferably the LY3300054 is administered at 70 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 700 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1000 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1400 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at a dose level needed to achieve constant saturation of available PD-L1 target protein by intravenous infusion during the course of combination therapy. Preferably the abemaciclib or the salt thereof is administered alone for 2 weeks of a first 28-day cycle and then starting on Day 15 of the first 28-day cycle and thereafter on Days 1 and 15 of subsequent cycles, the abemaciclib or the salt thereof is administered in combination with the LY3300054. Preferably on days the abemaciclib or the salt thereof and the LY3300054 are administered on the same day, the abemaciclib or the salt thereof is administered approximately 2 hours after starting the administration of the LY3300054.

According to a another aspect of the present invention, there is presented the use of LY3300054 in the manufacture of a medicament for the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein the LY3300054 is administered in simultaneous, separate, or sequential combination with abemaciclib, or a pharmaceutically acceptable salt thereof, after an initial administration of the abemaciclib or the salt thereof is administered without the LY3300054 for a time necessary to induce stable disease or partial response as determined by RECIST criteria or as determined by treating physician. Optionally, the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof and/or the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped after initiation of the combination with the LY3300054, and administration of the LY3300054 is continued without administering the abemaciclib or the salt thereof. Preferably the abemaciclib or the salt thereof is administered orally at 150 mg approximately every 12 hours. Preferably the abemaciclib or the salt thereof is administered orally at 100 mg approximately every 12 hours.

According to another aspect of the present invention, there is presented the use of

LY3300054 in the manufacture of a medicament for the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein the LY3300054 is administered in simultaneous, separate, or sequential combination with abemaciclib, or a pharmaceutically acceptable salt thereof, after an initial administration of the abemaciclib or the salt thereof is administered without the LY3300054 for a period of time which disease progression occurs as measured by RECIST criteria or as determined by treating physician.

Optionally, the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof and/or the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped after initiation of the combination with the LY3300054, and administration of the LY3300054 is continued without administering the abemaciclib or the salt thereof. Preferably the abemaciclib or the salt thereof is administered orally at 150 mg approximately every 12 hours. Preferably the abemaciclib is administered orally at 100 mg approximately every 12 hours. Preferably the

LY3300054 is administered at 70 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 700 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1000 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1400 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at a dose level needed to achieve constant saturation of available PD-L1 target protein by intravenous infusion during the course of combination therapy. Preferably the abemaciclib or the salt thereof is administered alone for 2 weeks of a first 28-day cycle and then starting on Day 15 of the first 28-day cycle and thereafter on Days 1 and 15 of subsequent cycles, the abemaciclib or the salt thereof is administered in combination with the LY3300054. Preferably on days the abemaciclib or the salt thereof and the LY3300054 are administered on the same day, the abemaciclib or the salt thereof is administered approximately 2 hours after starting the administration of the LY3300054.

According to another aspect of the present invention, there is presented the use of LY3300054 in the manufacture of a medicament for the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein the LY3300054 is administered in simultaneous, separate, or sequential combination with abemaciclib, or a pharmaceutically acceptable salt thereof, after an initial administration of the abemaciclib or the salt thereof is administered without the LY3300054 for a time necessary to induce intra-tumor immune inflammation. Optionally, the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof and/or the LY3300054. Optionally, the

According to another aspect of the present invention, there is presented the use of LY3300054 in the manufacture of a medicament for the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein the LY3300054 is administered in simultaneous, separate, or sequential combination with abemaciclib, or a pharmaceutically acceptable salt thereof, after an initial administration of the abemaciclib or the salt thereof is administered without the LY3300054 for a time necessary to increase expression of immune inflammatory genes. Optionally, the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof and/or the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped after initiation of the combination with the LY3300054, and administration of the LY3300054 is continued without administering the abemaciclib or the salt thereof. Preferably the abemaciclib or the salt thereof is administered orally at 150 mg approximately every 12 hours.

Preferably the abemaciclib or the salt thereof is administered orally at 100 mg for approximately 12 hours. Preferably the LY3300054 is administered at 70 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is

administered at 700 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1000 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1400 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at a dose level needed to achieve constant saturation of available PD-L1 target protein by intravenous infusion during the course of combination therapy. Preferably the abemaciclib or the salt thereof is administered alone for 2 weeks of a first 28-day cycle and then starting on Day 15 of the first 28-day cycle and thereafter on Days 1 and 15 of subsequent cycles, the abemaciclib or the salt thereof is administered in combination with the LY3300054. Preferably on days the abemaciclib or the salt thereof and the

According to another aspect of the present invention, there is presented the use of LY3300054 in the manufacture of a medicament for the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein the LY3300054 is administered in simultaneous, separate, or sequential combination with abemaciclib, or a pharmaceutically acceptable salt thereof, after an initial administration of the abemaciclib or the salt thereof is administered without the LY3300054 for a period of time necessary, prior to initiation of the combination, that the subsequent combination therapy of the abemaciclib or the salt thereof with the LY3300054 induces intra-tumor immune inflammation. Optionally, the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof and/or the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped after initiation of the combination with the LY3300054, and administration of the LY3300054 is continued without administering the abemaciclib or the salt thereof. Preferably the abemaciclib or the salt thereof is administered orally at 150 mg approximately every 12 hours. Preferably the abemaciclib or the salt thereof is administered orally at 100 mg approximately every 12 hours. Preferably the LY3300054 is administered at 70 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 700 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1000 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1400 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at a dose level needed to achieve constant saturation of available PD-L1 target protein by intravenous infusion during the course of combination therapy. Preferably the abemaciclib or the salt thereof is administered alone for 2 weeks of a first 28-day cycle and then starting on Day 15 of the first 28-day cycle and thereafter on Days 1 and 15 of subsequent cycles, the abemaciclib or the salt thereof is administered in combination with the LY3300054. Preferably on days the abemaciclib or the salt thereof and the LY3300054 are administered on the same day, the abemaciclib or the salt thereof is administered approximately 2 hours after starting the administration of the LY3300054. According to another aspect of the present invention, there is presented the use of abemaciclib, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control

(CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional tumors which display immune inflammation after a treatment with the compound or a salt thereof in a patient wherein the abemaciclib or the salt thereof is administered in simultaneous, separate, or sequential combination with LY3300054 after an initial administration of the abemaciclib or the salt thereof is administered without the LY3300054 for at least 24 hours. Optionally, the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof and/or the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped after initiation of the combination with the LY3300054, and administration of the LY3300054 is continued without administering the abemaciclib or the salt thereof. Preferably the abemaciclib or the salt thereof is administered orally at 150 mg approximately every 12 hours. Preferably the abemaciclib or the salt thereof is administered orally at 100 mg approximately every 12 hours. Preferably the LY3300054 is administered at 70 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 700 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1000 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1400 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at a dose level needed to achieve constant saturation of available PD-L1 target protein by intravenous infusion during the course of combination therapy. Preferably the abemaciclib or the salt thereof is administered alone for 2 weeks of a first 28-day cycle and then starting on Day 15 of the first 28-day cycle and thereafter on Days 1 and 15 of subsequent cycles, the abemaciclib or the salt thereof is administered in combination with the LY3300054. Preferably on days the abemaciclib or the salt thereof and the LY3300054 are administered on the same day, the abemaciclib or the salt thereof is administered approximately 2 hours after starting the administration of the LY3300054.

According to another aspect of the present invention, there is presented the use of abemaciclib, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein the abemaciclib or the salt thereof is administered in simultaneous, separate, or sequential combination with LY3300054 after an initial administration of the abemaciclib or the salt thereof is administered without the LY3300054 for at least 1 to 28 days.

According to another aspect of the present invention, there is presented the use of abemaciclib, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein the compound or salt thereof is administered in simultaneous, separate, or sequential combination with LY3300054 after an initial administration of the abemaciclib or the salt thereof is administered without the LY3300054 for a time necessary to induce stable disease or partial response as determined by RECIST criteria or as determined by treating physician. Optionally, the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof and/or the LY3300054. Optionally, the

According to another aspect of the present invention, there is presented the use of abemaciclib, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control

(CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein the abemaciclib or the salt thereof is administered in simultaneous, separate, or sequential combination with LY3300054 after an initial administration of the abemaciclib or the salt thereof is administered without the LY3300054 for a period of time which disease progression occurs as measured by RECIST criteria or as determined by treating physician. Optionally, the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof and/or the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped after initiation of the combination with the LY3300054, and administration of the LY3300054 is continued without administering the abemaciclib or the salt thereof. Preferably the abemaciclib or the salt thereof is administered orally at 150 mg approximately every 12 hours. Preferably the abemaciclib or the salt thereof is administered orally at 100 mg approximately every 12 hours.

According to another aspect of the present invention, there is presented the use of abemaciclib, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein the abemaciclib or the salt thereof is administered in simultaneous, separate, or sequential combination with LY3300054 after an initial administration of the abemaciclib or the salt thereof is administered without the LY3300054 for a time necessary to induce intra-tumor immune inflammation. Optionally, the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof and/or the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped after initiation of the combination with the LY3300054, and administration of the LY3300054 is continued without administering the compound, or the salt thereof. Preferably the abemaciclib or the salt thereof is administered orally at 150 mg approximately every 12 hours. Preferably the abemaciclib or the salt thereof is administered orally at 100 mg approximately every 12 hours. Preferably the LY3300054 is administered at 70 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 700 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1000 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1400 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at a dose level needed to achieve constant saturation of available PD-L1 target protein by intravenous infusion during the course of combination therapy. Preferably the abemaciclib or the salt thereof is administered alone for 2 weeks of a first 28-day cycle and then starting on Day 15 of the first 28-day cycle and thereafter on Days 1 and 15 of subsequent cycles, the abemaciclib or the salt thereof is administered in combination with the LY3300054. Preferably on days the abemaciclib or the salt thereof and the

According to another aspect of the present invention, there is presented the use of abemaciclib, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma such as mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein the medicament is administered in simultaneous, separate, or sequential combination with an antibody which is LY3300054 after an initial administration of the abemaciclib or the salt thereof is administered without the LY3300054 for a time necessary to increase expression of immune inflammatory genes. Optionally, the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate adverse effects of the abemaciclib or the salt thereof and/or the LY3300054. Optionally, the administration of the abemaciclib or the salt thereof is stopped after initiation of the combination with the LY3300054, and administration of the LY3300054 is continued without administering the abemaciclib or the salt thereof. Preferably the abemaciclib or the salt thereof is administered orally at 150 mg approximately every 12 hours. Preferably the abemaciclib or the salt thereof is administered orally at 100 mg approximately every 12 hours. Preferably the LY3300054 is administered at 70 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 700 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1000 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1400 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at a dose level needed to achieve constant saturation of available PD-L1 target protein by intravenous infusion during the course of combination therapy. Preferably the abemaciclib or the salt thereof is administered alone for 2 weeks of a first 28-day cycle and then starting on Day 15 of the first 28-day cycle and thereafter on Days 1 and 15 of subsequent cycles, the abemaciclib or the salt thereof is administered in combination with the LY3300054. Preferably on days the abemaciclib or the salt thereof and the LY3300054 are administered on the same day, the abemaciclib or the salt thereof is administered approximately 2 hours after starting the administration of the LY3300054.

(CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein the abemaciclib or the salt thereof is administered in simultaneous, separate, or sequential combination with an antibody which is LY3300054 after an initial administration of the abemaciclib or the salt thereof is administered without the

LY3300054 for at least 24 hours. Preferably the abemaciclib or the salt thereof is administered orally at 150 mg approximately every 12 hours. Preferably the abemaciclib or the salt thereof is administered at 100 mg approximately every 12 hours. Preferably the LY3300054 is administered at 70 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 700 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1000 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1200 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at 1400 mg once every two weeks by intravenous infusion. Preferably the LY3300054 is administered at a dose level needed to achieve constant saturation of available PD-L1 target protein by intravenous infusion during the course of combination therapy. Preferably the abemaciclib or the salt thereof is administered alone for 2 weeks of a first 28-day cycle and then starting on Day 15 of the first 28-day cycle and thereafter on Days 1 and 15 of subsequent cycles, the abemaciclib or the salt thereof is administered in combination with the LY3300054. Preferably on days the abemaciclib or the salt thereof and the LY3300054 are administered on the same day, the abemaciclib or the salt thereof is administered approximately 2 hours after starting the administration of the LY3300054.

For the above aspects, preferably, the compound or salt thereof is in combination with the LY3300054 for the treatment of NSCLC. Preferably, abemaciclib or the salt thereof is in combination with the LY3300054 for the treatment of breast cancer.

Preferably, the abemaciclib or the salt thereof is in combination with the LY3300054 for the treatment of pancreatic cancer. Preferably, the abemaciclib or the salt thereof is in combination with the LY3300054 for the treatment of mantle cell lymphoma. Preferably, the abemaciclib or the salt thereof is in combination with the LY3300054 for the treatment of tumors which display immune inflammation after a treatment with the abemaciclib or the salt thereof. Preferably, the abemaciclib or the salt thereof is in combination with the LY3300054 for the treatment of tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway).

As used herein, the term "anti" when used with a target and the term antibody, such as "anti"-PD-l antibody means an antibody or antibodies that block interactions between the target and its ligands.

As used herein, the term "initial administration" means the first time that abemaciclib, or a pharmaceutically acceptable salt thereof, is administered to a patient as part of the present invention.

As used herein, the term "additional susceptible tumors" means tumors, which display immune inflammation after a treatment with abemaciclib or the salt thereof, in a patient.

As used herein, "LY3300054" is an antibody that binds human PD-L1 (SEQ ID NO: 1), comprising a light chain (LC) and a heavy chain (HC), wherein the light chain comprises a light chain variable region (LCVR) and the heavy chain comprises a heavy chain variable region (HCVR), and wherein the LCVR comprises light chain

complementarity determining regions LCDR1, LCDR2, and LCDR3, where the amino acid sequence of LCDR1 is SGSSSNIGSNTVN (SEQ ID NO: 5), the amino acid sequence of LCDR2 is YGNSNRPS (SEQ ID NO: 6), and the amino acid sequence of LCDR3 is QSYDSSLSGSV (SEQ ID NO: 7), and wherein the HCVR comprises heavy chain complementarity determining regions HCDR1, HCDR2, and HCDR3, where the amino acid sequence of HCDR1 is KASGGTFS S YAIS (SEQ ID NO: 2), the amino acid sequence of HCDR2 is GIIPIFGTANYAQKFQG (SEQ ID NO: 3), and the amino acid sequence of HCDR3 is ARSPDYSPYYYYGMDV (SEQ ID NO: 4), respectively.

In some embodiments of LY3300054, the LY3300054 binds to human PD-L1, and comprises a light chain (LC) and a heavy chain (HC), wherein the light chain comprises a light chain variable region (LCVR) and the heavy chain comprises a heavy chain variable region (HCVR), wherein the amino acid sequence of the LCVR is SEQ ID NO: 9, and the amino acid sequence of the HCVR is SEQ ID NO: 8. In some embodiments of LY3300054, the LY3300054 binds to human PD-L1, comprising a light chain (LC) and a heavy chain (HC), wherein the amino acid sequence of the LC is SEQ ID NO; 10 and the HC has the amino acid sequence given in SEQ ID NO: 11. In an embodiments of LY3300054, the LY3300054, comprises two light chains and two heavy chains, wherein each light chain has the amino acid sequence given in SEQ ID NO: 11, and each heavy chain has the amino acid sequence given in SEQ ID NO: 10.

As used herein, the term "light chain variable region" or "LCVR" means a portion of a light chain of an antibody molecule that includes amino acid sequences of CDRs and FRs.

As used herein, the term "heavy chain variable region" "HCVR" means a portion of a heavy chain of an antibody molecule that includes amino acid sequences of CDRs and FRs.

As used herein, the terms "complementarity determining region" and

"CDR", mean the non-contiguous antigen combining sites found within the variable region of LC and HC polypeptides of an antibody or an antigen-binding fragment thereof. These particular regions have been described by others including Kabat, et al., Ann. NY Acad. Sci. 190:382-93 (1971); Kabat et al., J. Biol. Chem. 252:6609- 6616 (1977); Kabat, et al., Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91- 3242 (1991); Chothia, et al., J. Mol. Biol. 196:901-917 (1987); MacCallum, et al., J. Mol. Biol., 262:732-745 (1996); and North, et al., J. Mol. Biol., 406, 228-256 (2011), where the definitions include overlapping or subsets of amino acid residue when compared against each other.

The CDRs are interspersed with regions that are more conserved, termed framework regions ("FR"). Each LCVR and HCVR is composed of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FRl, CDR1, FR2, CDR2, FR3, CDR3, FR4. The three CDRs of the light chain are referred to as "LCDR1, LCDR2, and LCDR3" and the three CDRs of the heavy chain are referred to as "HCDR1, HCDR2, and HCDR3." The CDRs contain most of the residues which form specific interactions with the antigen. The numbering and positioning of CDR amino acid residues within the LCVR and HCVR regions is in accordance with known conventions (e.g., Kabat (1991), Chothia (1987), and/or North (2011)). In different embodiments of the invention, the FRs of the LY3300054 may be identical to the human germline sequences, or may be naturally or artificially modified.

As used herein, the term "KD" means the equilibrium dissociation constant of a particular antibody- antigen or antibody fragment-antigen interaction.

As used herein, the term "binds" means the affinity of an antibody for human PD- Ll is intended to mean, unless indicated otherwise, a KD of less than about 1 xlO ⁶ M, preferably, less than about 1 x 10^~9 M as determined by common methods known in the art, including by use of a surface plasmon resonance (SPR) biosensor at 37°C essentially as described herein.

As used herein, the term "kit" means 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 LY3300054. 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.

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

As used herein, the term "patient" means to a mammal, preferably a human. As used herein, the term "cancer" means or describes the physiological condition in patients that is typically characterized by unregulated cell proliferation. Included in this definition are benign and malignant cancers.

As used herein, the term "primary tumor" means the original cancer and not a metastatic lesion located in another tissue, organ, or location in the subject's body.

As used herein, the term "effective amount" means the amount or dose of abemaciclib, or a pharmaceutically acceptable salt thereof, and the amount or dose of LY3300054 which provides an effective response in the patient under diagnosis or treatment. 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 refers to the clinical or therapeutic benefit imparted to a patient upon administration of abemaciclib, or a pharmaceutically acceptable salt thereof, and LY3300054.

As used herein, the term "abemaciclib" means a CDK4/6 inhibitor disclosed in WO2010/075074 with the following structure:

The free base of abemaciclib is preferred. However, it will be understood by the skilled reader that 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, et al.,

Encyclopedia of Pharmaceutical Technology, 453-499 (1995); S.M. Berge, et al., Journal of Pharmaceutical Sciences, 66, 1, (1977). The hydrochloride and mesylate salts are preferred salts. The mesylate salt is an especially preferred salt.

Abemaciclib or pharmaceutically acceptable salts thereof, may be prepared by a variety of procedures known in the art (e.g., see WO2010/075074).

Dosages per day of abemaciclib, or a pharmaceutically acceptable salt thereof, normally fall within the range of 50 mg to 200 mg twice daily, more preferably 100-150 mg twice daily. Most preferably 100 mg twice daily. Also most preferable 150 mg twice daily.

It is understood that a combination therapy of the present invention is carried out by administering LY3300054 together with a compound that is abemaciclib, or a pharmaceutically acceptable salt thereof, in any manner which provides effective levels of the LY3300054 and/or the compound or salt thereof.

It is understood that the time necessary to induce intra-tumor immune

inflammation or the time necessary to increase expression of immune inflammatory genes can be measured many ways, for example, by RNA gene expression analysis methods, such as PCR, quantitative PCR, QUANTIGENE®, nanostring, RNA-sequencing, gene microarray and Modeplex, preferably nanostring.

The route of administration of the abemaciclib, or the salt thereof or LY3300054 may be varied in any way, limited by the physical properties of the drugs and the convenience of the patient and the caregiver. Preferably, the LY3300054 is formulated for parenteral administration, such as intravenous or subcutaneous administration.

Preferably, compound and the salt thereof are formulated for oral or parenteral administration, including intravenous or subcutaneous administration.

In particular, the compound or the salts thereof may be formulated into a tablet or capsule. Such pharmaceutical compositions and processes for preparing same are well known in the art. (See, e.g., Remington: The Science and Practice of Pharmacy, L.V. Allen, Editor, 22^nd Edition, Pharmaceutical Press, 2012). The antibody or pharmaceutical compositions comprising the same may be administered by parenteral routes (e.g., subcutaneous and intravenous). An antibody of the present invention may be

administered to a patient alone with pharmaceutically acceptable carriers, diluents, or excipients in single or multiple doses. Pharmaceutical compositions of the present invention can be prepared by methods well known in the art (e.g., Remington: The Science and Practice of Pharmacy, 22^nd ed. (2012), A. Loyd et al., Pharmaceutical Press) and comprise an antibody, as disclosed herein, and one or more pharmaceutically acceptable carriers, diluents, or excipients.

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, overall 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, may induce shrinkage of the primary tumor, or may simply exert a tumoristatic effect. Because the invention relates to the use of a combination of unique anti-tumor agents, novel approaches to determining efficacy of any particular combination therapy of the present invention can be optionally employed, including, for example, measurement of plasma or urinary markers of angiogenesis and/or cell cycle activity, tissue-based biomarkers for angiogenesis and/or cell cycle activity, and measurement of response through radiological imaging. Experimental Rationale for Examples 1-5

To study the combination potential of CDK4/6 inhibitors, such as abemaciclib, with inhibitors of a co-inhibitory immune checkpoint and/or activators of co- stimulatory immune checkpoints (Checkpoint Immunotherapy) , such as an anti-PD-Ll antibody that blocks interaction with its ligands (anti-PD-Ll, or PD-Ll therapy), monotherapy and combination therapy experiments are performed in the CT26 syngeneic tumor model. The CT26 syngeneic tumor model is a chemically induced murine tumor from Balb/C mice for which gene expression analysis has predicted it to be sensitive to cell cycle inhibitors (Castle, et al , BMC genomics 2014). This tumor harbors genetic disruptions in cell cycle control such as CDKN2a deletion and expresses a functional Retinoblastoma protein (Rb), making it potentially sensitive to growth inhibition by abemaciclib (Finn et al. Breast Cancer Research 2009, 11:R77).

CT26 is also a well-known model for immunotherapy. Studies have shown it to be sensitive to inhibitors of a co-inhibitory immune checkpoint including, CTLA-4, PD-1, PD-Ll blockade, and activators of co-stimulatory immune checkpoint of CD137, GITR, and OX40. Established sensitivity of CT26, and well known range of therapeutic efficacy of immunotherapeutic intervention make it an appropriate model for screening the activity of checkpoint immunotherapies (Grosso and Jure-Kunkel, Cancer Immunity Vol. 13, p. 5, Jan 2013) (Duraiswamy et al, Cancer Research, June 15, 2013 73;3591) .

CT26 is a murine model, and accordingly, antibodies used in the CT26 model must bind the mouse target protein. To model co-inhibitory immune checkpoint therapy, in particular blockade of PD-Ll/PD-1 mediated inhibition, anti-murine PD-Ll clone, 178G7 (Known herein in the examples as: anti-PD-Ll antibody or anti-PD-Ll therapy or anti-PD-Ll), is produced. The HC of 178G7 is SEQ ID NO: 14, and the LC of 178G7 is SEQ ID NO: 15. 178G7 blocks PD-1 binding to PD-Ll. In particular, 178G7 is a surrogate for LY3300054, as both 178G7 and LY3300054 block binding of PD-1 to PD- Ll, and PD-Ll to CD80. 178G7 competes with previously identified surrogate antibody 10F.9G2 known to block PD-Ll/PD-1 interaction and is a known surrogate for anti- human antibodies in the clinic (Eppihimer et al. Microcirculation 2002:9(2): 133).

Example 1 CT26 demonstrates sensitivity to CDK4/6 inhibitors

To determine if CT26 displays in vitro sensitivity to CDK4/6 inhibitors, CT26 cells (American Type Culture Collection) are plated 1000 cells/well in 100 media (RPMI 1640+10% FBS) in a 96 well plate. Cells are cultured for 4 hours alone at 37°C and then abemaciclib, palbociclib or vehicle control (dimethyl sulfoxide (DMSO)) are added in duplicate wells at final concentrations from 20 μΜ to 0.001 μΜ (ten point titration in 1:3 dilution steps, Table 2). Cells are cultured for 96 hours at 37°C and then assessed for cell viability using CELLTITER-GLO® (Promega). A volume of

CELLTITER-GLO® reagent (lyophilized CELLTITER-GLO® substrate reconstructed with CELLTITER-GLO® buffer) equal to the medium in the well, is then added to each well of the 96-well plate. Samples are placed onto an orbital shaker to induce cell lysis and then incubated at room temperature for 10 minutes. Luminescence is then recorded on a Perkin Elmer Wallac Victor 2 plate reader. Percent viability inhibition is shown in Table 2 with viability calculated according to the formula: (l-(mean luminosity of treated sample/mean luminosity of untreated control)) x 100. Mean values are then used to calculate a 50% inhibitory concentration (IC50) for growth or viability inhibition using four parameter logistic curve fit formula. Calculation of IC50 shows that abemaciclib inhibits CT26 cell growth or viability with an IC50 of 2.7 μΜ and palbociclib with an IC50 of 4.5 μΜ.

Table 2: In Vitro Viability Inhibition Of CT26 Cells

The above example demonstrates that growth of CT26 cancer cells can be effectively inhibited by CDK4/6 inhibitors and thereby represents a model to test activity of these inhibitors in combination with checkpoint immunotherapy. Example 2

Abemaciclib inhibits CT26 tumor growth as monotherapy and induces intra tumor immune inflammation

To determine the effects of abemaciclib on the growth of CT26 syngeneic tumors, CT26 tumor cells are cultured for implantation in medium (RPMI1640; 10% fetal bovine serum; 1 mM sodium pyruvate; 2 mM L-glutamine). Cells are passaged twice a week before harvesting and then Balb/C mice are implanted in the rear flank with 1 x 10⁶ CT26 cells in phosphate buffered saline. Tumors are allowed to grow for 6 days to between 50- 100 mm³ in size before treatment. At 6 days following implantation (study day 6) groups of 15 animals are treated with either vehicle (1% hydroxy ethyl cellulose (HEC) in 25 mM phosphate buffer, pH 2 ) 25 mg/kg of abemaciclib, 50 mg/kg of abemaciclib or 75 mg/kg of abemaciclib. Treatment is administered orally, once a day for 21 days. 5 animals per group remain in the study and are followed until the end of the study as noted below, to measure the durability of efficacy compared to control. To investigate intra-tumor effects on immune inflammation resulting from treatment, subgroups of 5 animals from each treatment group are sacrificed on study day 13 and study day 20 for mechanism of action analysis (MO A). Tumors and spleens are collected from each of these 5 animals and are analyzed for percentage of immune cell subsets listed below by flow cytometry and RNA is prepared from a portion of the remaining tumor to measure immune gene expression during therapy.

If tumor burden becomes greater than 2500 mm³ animals are sacrificed due to progressive disease, or at lower volume if the size of the tumor suggests that they will surpass 2500 mm³ before the next scheduled measurement in accordance with animal use policies. On study day 84, any mice that achieved complete regression (tumor volume < 25 mm³) of their tumors are rechallenged with CT26 tumors on the opposite flank of the original tumor injection site. Tumor growth of the secondary challenge tumor is then followed for 22 days to measure the acquisition of immunologic memory. Tumor growth analysis

Shown in Table 3, abemaciclib treatment resulted in a dose dependent inhibition of tumor growth showing that CT26 model is sensitive to CDK4/6 inhibition. Tumor volume compared to control are analyzed at day 19, at the point where 10 animals are present in each group. Ratio of tumor volume compared to control (%T/C) is calculated using the formula %T/C = 100 x T/C, whereby T = 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 C = mean tumor volume of the control group minus the mean tumor volume on the baseline day. The baseline day is the nearest day just prior to the initiation of treatment or the first measurement day of the study. Tumor volume data is then analyzed with a two-way repeated measures analysis of variance by time and treatment, the response was analyzed using the log transformation of tumor volume. Predefined pairwise comparisons of treated group(s) to control group for the indicated time point were conducted.

Treatment resulted in a dose dependent tumor growth inhibition at all doses tested (25 mg/kg %T/C= 58%, 50 mg/kg %T/C=38.9%, 75 mg/kg %T/C=27.9% ) whereby tumor growth inhibition observed at both 50 mg/kg and 75 mg/kg were statistically significantly from the control (Table 3). From the 5 animals which remained in the study for each group, after collection of MOA animals, abemaciclib caused complete regression (CR=tumor volume < 25 mm³) in 2 of the 5 remaining animals in the 75 mg/kg group. No CRs were achieved in other groups. The CR animals were re- challenged by implanting additional CT26 tumor cells on the opposite flank. These animals resisted this secondary challenge as indicated by the inability of these newly implanted CT26 cells to grow to form measurable tumors.

Table 3: Monotherapy Efficacy Of Abemaciclib Compared To Control In CT26 Tumors.

-52-

%T/C= Mean tumor volume of treatment group / mean tumor volume of control , CR= # of animals reaching complete regression of tumor volume below 25 mm³

Cell subset frequency analysis by fluorescence-activated cell sorting (FACS)

Spleens and Tumors are collected from MOA animals on study day 13 and study day 20 as described above and processed for FACS analysis by preparing a single cell suspension in tissue culture medium. Single cell suspensions are then incubated with fluorescently labeled antibodies to identify immune markers; CD3, CD4, CD8, CD l ib, CD 19, CD45, FoxP3, and a fixable viability dye (Affymetrix). Labeled single cell suspensions of tissues are then collected individually from each mouse are then analyzed using a flow cytometer. Data is collected and then analyzed using Flowjo (Treestar) to measure the percentage of total live lymphocytes (Live, CD45+) in each tissue collected from each animal. Immune cell subsets are then measured as a percentage of CD45+ cells.

Shown in Table 4, monotherapy with abemaciclib caused minor but significant alterations of immune cell subset frequency in spleen at both study day 13 and study day 20, with most significant changes centered around a reduction of myeloid cells

(CD45+CDl lb+) and increase in T cells (CD45+,CD3+).

Table 4: Intra-Splenic Immune Cell Frequencies

-53-

-54-

*= P < 0.05 compared to vehicle control (one way Anova, fishers LSD)

Intra-tumor immune gene expression

To investigate the effects of abemaciclib treatment on the intra-tumor microenvironment, tumor tissue is snap frozen in liquid N2 after isolation from mice at the MOA timepoints. Total RNA is isolated from lysed samples using the MAGMAX™ 96 Total RNA isolation kit (Life Technologies). Snap-frozen tumors are lysed by adding 500 μί, of prepared lysis/binding buffer (MAGMAX™ 96 Total RNA isolation kit) with a single stainless steel beads (5 mm ) and then homogenized on a TissueLyser (Qiagen) for 2 minutes at 25Hz. 200 μί. of lysate, along with 120 μί. of 100% isopropanol and 40 μΐ_^ of prepared magnetic beads mix is then transferred to a 96 deep well processing plate. A series of washes and a DNase incubation step are processed on the MAGMAX™ Express -96 Deep Well Magnetic Particle Processor. RNA is eluted from the magnetic beads with 100 μί. of elution buffer after a final wash step. RNA concentration is determined spectrophotometrically to determine optical density at 260 and 280 nm and is then diluted to the final concentration of 25 ng/uL. Tissue expression of immune related genes (Table 6) is then measured using QUANTIGENE® 2.0 plexassay (Affymetrix).

Table 6 - Immune Related Gene Probe Set

500 ng of total RNA from samples being analyzed is added in duplicate to individual wells of a 96-well hybridization plate containing QUANTIGENE® magnetic capture beads, QUANTIGENE® probesets, and QUANTIGENE® blocking reagent for a final volume of 100 μΐ,. The hybridization plate is then sealed and incubated at 54°C while agitated at 600 rpm for 16 hours. Samples are then transferred to a 96-well magnetic plate and beads are then sequentially hybridized first with QUANTIGENE® Pre- Amplifier Probe, then, with QUANTIGENE® Amplifier Probe and finally with

QUANTIGENE® Label Probe (1 hour at 50°C for each step, with washing between each step). Strep tavidin-conjugated R-Phycoerythrin (SAPE, Affymetrix) is added after the last hybridization step and incubated for 30 minutes at room temperature. Samples are then analyzed on the FLEXMAP 3D® Luminex instrument (ThermoFisher, Waltham, MA) and level of RNA detection is determined by measuring the mean fluorescence intensity (MFI) level of Phycoerythrin detected. MFI data is then then converted into relative gene expression for each gene (Normalized Adjusted Net MFI) using a quality control analysis script developed in-house to perform the following calculations for each gene measure for each sample by calculating the "Net MFI" for each gene for each sample, (Sample MFI - background MFI of blank well). Next the lower limit of detection (LLOD) is determined (background MFI + 3 standard deviation (SD)). The LLOD is then used to calculate the "Adjusted Net MFI" (if MFI > LLOD, than "Adjusted Net MFI" = "Net MFI", if MFI < LLOD, than "Adjusted Net MFI" = LLOD - background). Relative gene expression is then calculated by normalizing for each gene's adjusted net MFI to the geometric mean of the MFI of selected housekeeping genes (HKG) including Hptrl, Gusb, Rpsl8 and Ppib shown in Table 6 (Adjusted Net

MFI/geometric mean HKG MFI) multiplied by a scaling factor of 100. Fold change in gene expression for each gene for each sample in each treatment group is then determined compared to the control group using the following formula (Normalized Adjusted Net MFI Treatment sample/ Mean Normalized adjusted Net MFI), fold change values <1 = negative fold change, fold change values 1< = positive fold change. Statistical analysis of the mean fold change for each gene of each treatment group analyzed is then performed compared to control using two-way ANOVA tests to determine the significance of the difference between groups. Fold change of abemaciclib treated compared to vehicle control treated mice is shown in Table 7.

Although abemaciclib targets CDK4/6, unexpectedly, treatment also causes a dose dependent increase in immune inflammation as indicated by increases in the expression of multiple immune related genes compared to untreated animals. Genes that are significantly changed include genes encoding T cell activation markers [IFNy (Ifng), granzyme B (Gzmb)], co-stimulatory/co-inhibitory receptors and/or ligands [CD 137 (Tnfrsf9), PD-L1 (Cd274), PD-L2 (Pdcdllg2), TIM-3 (Havcr2)], chemokines [MIPlcc (Ccl3), ΜΙΡΙβ (Ccl4), RANTES (Ccl5), GRO (Cxcll )] and immunosuppressive enzymes (Idol ) that might be indicative of IFNy-induced T cell activation and inflammatory response. Abemaciclib monotherapy is associated with transient downregulation of arginase (Argl) in CT26 tumors.

Table 7: Abemaciclib Causes Dose And Time Dependent Induction Of Intra-Tumor Immune Inflammation

change values 1< = positive

Data presented in this example, and Example 3, 4& 5 below, supports that CT26 is a murine tumor that is sensitive to CDK4/6 inhibition as indicated by the dose dependent suppression of tumor growth observed following treatment with abemaciclib (Example 2, 3,4,5) or palbociclib (Example 5). The sensitivity of CT26 to abemaciclib demonstrates that this is an appropriate model for determining similar activity of CDK4/6 inhibitors in human tumors. This data also supports the conclusion that abemaciclib monotherapy causes changes in the intra-tumor immune microenvironment leading to increases in inflammatory immune gene expression and increase expression of immune activation markers. The dose dependent increase in intra- tumoral inflammatory signature which correlated with immune mediated complete regression at the highest doses of abemaciclib suggested that abemaciclib in addition to being cell cycle inhibitor, also has the potential to be a modulator of intra-tumor immune function.

Example 3

Concurrent dosing of abemaciclib with PD-Ll checkpoint immunotherapy results in equivalent anti-tumor efficacy compared to abemaciclib monotherapy

Abemaciclib is combined concurrently from the initiation of therapy with anti-PD- Ll checkpoint immunotherapy to see if it can improve abemaciclib monotherapy. This would mimic a potential clinical setting where an anti-PD-Ll antibody therapy would be used to improve the response to abemaciclib. The concurrent combination efficacy experiment in CT26 tumors is run essentially as described in Example 2, with the following modifications:

To determine the ability of an anti-PD-Ll checkpoint immunotherapy to enhance the efficacy of abemaciclib, 50 mg/kg of abemaciclib is tested in combination with an anti-PD-Ll antibody; Starting 6 days after tumor challenge, groups of 5-10 mice are treated with either vehicle, 50 mg/kg abemaciclib for 21 days in monotherapy, anti-PD- Ll antibody therapy alone (178G7 flat dosed 500 μg/dose 1 time per week, for 3 doses sufficient to saturate PD-Ll for 3 weeks), or a concurrent combination of 50 mg/kg abemaciclib for 21 days combined with 3 weekly doses of the anti-PD-Ll antibody therapy Tumors are followed as described above in Example 2.

Tumor growth analysis

As shown in Table 8, abemaciclib treatment caused a inhibition of tumor growth similar to showing that CT26 model is sensitive to CDK4/6 inhibition. Relative changes in tumor volume compared to control are analyzed at day 24, at the point where the most animals remained in all groups for proper statistical comparison. Treatment with abemaciclib caused a significant tumor growth inhibition in monotherapy compared to control (%T/C=22.3%). Anti-PD-Ll antibody monotherapy treatment also resulted in a growth delay compared to control, which trended toward significance (%T/C=56.1). Concurrent combination of abemaciclib with the anti-PD-Ll antibody caused a significant growth delay compared to control (%T/C=25.4%) however this was not significantly different from abemaciclib monotherapy. No CRs were achieved in this study

Table 8: Concurrent Combination Efficacy Of Abemaciclib And An Anti-PD-Ll Antibody Compared To Control In CT26 Tumors.

In the above example, concurrent dosing from the initiation of therapy of both abemaciclib and the anti-PD-Ll results in improved effacacy compared to PD-L1 monotherapy, but there was no apparent difference in the anti-tumor response compared to abemaciclib alone. This data demonstrates that abemaciclib can be added to PD- Ll/PD-1 blockade therapy without reducing the efficacy of PD-Ll/PD-1 blockade, and supports that cell cycle inhibitors can be combined with immunotherapy checkpoint blockade.

Example 4

Short term overlapped combination of abemaciclib with PD-L1 checkpoint immunotherapy results in increased anti-tumor efficacy compared to monotherapy

To determine if it is possible to counteract potential adverse effects on T cells, and provide sufficient time for the positive immune effects of abemaciclib to occur, abemaciclib is combined either sequentially, or with short concurrent overlap with checkpoint immunotherapy to see if this could potentially result in a synergistic effect.

To determine the ability of abemaciclib to enhance or synergize with checkpoint immunotherapy, in this case an anti-PD-Ll antibody, a lower dose of 50 mg/kg abemaciclib is chosen to enhance the detection of a possible combinatorial effect. The combination efficacy experiment in CT26 tumors is run essentially as described in Example 2, with the following modifications; Groups of 9-10 mice are treated, starting 6 days after tumor challenge with vehicle, 50 mg/kg abemaciclib for 14 days in

monotherapy, 50 mg/kg abemaciclib for 14 days followed by an anti-PD-Ll antibody therapy sufficient to saturate PD-L1 for 3 weeks (antibody clone 178G7 as described above, flat dosed 500 μg/dose 1 time per week, for 3 doses) starting on day 21,

(sequential combination), or 50 mg/kg abemaciclib treatment starting on day 6 for a total 14 days, which is given for 7 days as a monotherapy prior to the initiation of combination therapy with an anti-PD-Ll antibody (178G7 flat dosed 500 μg/dose 1 time per week, for 3 doses) beginning on day 13 (1 week overlap combo), in combination with an anti-PD- Ll antibody alone beginning on day 6 (178G7 flat dosed 500 μg/dose 1 time per week, for 3 doses). Tumors are followed as described above in Example 2 with the study completed on study day 64.

Tumor growth analysis

To measure the median survival of animal after therapy, Kaplan- Meier analysis method is performed on the tumor growth data to obtain log-rank p- values for the comparison of groups (Table 9). A parametric survival analysis method is used to test the interaction effect of each combination groups to determine synergy, additivity, less than additivity, or antagonism in survival.

For Kaplan-Meier analysis method: The time to sacrifice is determined for each animal. If the final tumor volume is 25 mm³ or less, the animal is considered a complete responder (CR) and the time is recorded as right-censored. Predicted median survival times and standard errors are derived from this model. A t-test for interaction effect is performed on the median survival times to compare the combination group versus monotherapies to determine synergy versus additive responses.

Shown in Table 9, abemaciclib combination treatment resulted in a significant, regimen specific increase in median survival as compared to abemaciclib monotherapy or 178G7 monotherapy. 14 days of abemaciclib monotherapy caused a measureable inhibition in tumor growth, and anti-PD-Ll antibody monotherapy only caused a trend towards increased time of survival as compared to control, however this was not statistically significant. Sequential combination treatment with an anti-PD-Ll antibody and abemaciclib resulted in significant increase in time of survival as compared to control, but was not significantly different from the monotherapies. The one week overlap combination group showed the most tumor growth inhibition and the greatest increase in survival compared to every other group, with 2 animals achieving complete responses. The increase in survival seen with 1 week overlap combination was determine to be statistically significant compared to controls, monotherapies, and additionally was shown to be synergistic as compared to the responses seen with monotherapy abemaciclib or anti-PD-Ll antibody therapy.

Table 9: Kaplan Kaplan-Meier , Parametric Survival Analysis Of Tumor Survival For Combination Efficacy Of Abemaciclib With Anti-PD-Ll Antibody In CT26 Tumors

In the above example, abemaciclib demonstrated combinatorial potential with checkpoint immunotherapy. Both sequential therapy and pretreatment with abemaciblib before a short overlap of combination therapy demonstrated superior tumor growth inhibition as compared to anti-PD-Ll antibody monotherapy group. The 1 week overlap combination showed singinifant increase in survival compared to abemacilcib monotherapy, and the combination was found synergistic compared to the monotherapy groups.

Example 5

Immune modulating potential of Abemaciclib enhances the efficacy of CDK4/6 inhibitor combinations with checkpoint immunotherapy in a dose dependent manner.

Example 4 demonstrates that a short overlap of abemaciclib therapy in combination with checkpoint immunotherapy could enhance tumor growth inhibition. Additionally, this example demonstrated that the scheduling of abemaciclib relative to anti-PD-Ll antibody therapy may be an important aspect for achieving optimal anti- tumor efficacy whereby pretreatment with abemaciclib followed by an overlap of combination therapy with anti-PD-Ll antibody therapy and abemaciclib may be superior to schedules that dose abemaciclib and anti-PD-Ll antibody in a completely sequential and non-overlapping fashion. To extend on this discovery, further studies are initiated aimed at evaluating whether better combination efficacy could be achieved by increasing the dose of abemaciclib. Since abemaciclib clinical dosing is continuous, without stopping , it is also important to understand if a longer combination overlap, after an initial phase of abemaciclib monotherapy, would also enhance checkpoint immunotherapy. Finally, in this example, we compare the efficacy of abemaciclib to palbociclib, to understand if other CDK4/6 inhibitors have similar effects in combination with checkpoint immunotherapy. To address these questions, mice are implanted with CT26 tumors as described in Examples 2, 3 and 4 with the following modifications as described below.

To determine monotherapy efficacy, starting after 6 days of tumor growth, groups of 10 mice are treated with vehicle control, anti-PD-Ll antibody, 50 mg/kg of abemaciclib, 75 mg/kg of abemaciclib, or 75 mg/kg of palbociclib. Both abemaciclib and palbociclib are dosed for 28 days to evaluate the effects of a longer duration of monotherapy therapy and compare to extended combination therapy.

To determine the efficacy of a 1 week combination overlap (1 week overlap), groups of 10 mice are treated using the schedule described in Example 4, and are given once-daily monotherapy with either 50 mg/kg of abemaciclib, 75 mg/kg of abemaciclib, or 75 mg/kg of palbociclib for 14 days, starting 6 days after tumor implantation and then given these same doses of the CDK4/6 inhibitors for 3 additional weeks in combination with once-weekly anti-PD-Ll antibody therapy starting on study day 13.

To determine the efficacy of extended combination overlap (extended combo) groups of 10 mice are treated once-daily with either 50 mg/kg of abemaciclib, 75 mg/kg of abemaciclib, or 75 mg/kg of palbociclib for 28 days, starting 6 days after tumor implantation with anti-PD-1 antibody therapy once-weekly starting on day 13, with the last of 3 doses given on study day 27, 7 days before the cessation of treatment with the CDK4/6 inhibitors.

In parallel to the efficacy study, groups of 5 animals are treated with the same monotherapy abemaciclib and palbociclib doses and combination therapies as described above and sacrificed for MOA analysis on study day 20, 24 hours after the last dose of abemaciclib and palbociclib, and before dosing with the anti-PD-Ll antibody where applicable. An additional 5 mice are treated with one dose of the anti-PD-Ll antibody in monotherapy on study day 13 to serve as an MOA control for the combination groups and collected at study day 20. Pooled spleens, tumors and plasma are collected for MOA analysis as described in Example 2.

Measurements of body weights and tumor volumes from all animals in the above groups are captured routinely during the course of the experiment as described above in Example 2. Primary tumor growth is followed until study day 69 at which point any animal demonstrating a complete efficacy response is rechallenged on the opposite flank with CT26 as described in Example 2. The secondary challenge phase is then followed for two weeks to assess for possible tumor formation.

Primary tumor growth analysis

To quantitate the responses seen, statistical analysis is performed as described in

Example 3. Shown in Table 9, all combinations demonstrated improved median survival as compared to controls, and anti-PD-Ll antibody monotherapy. The interaction effect of 50 mg/kg abemaciclib with anti-PD-Ll antibody therapy was determined to be synergistic for both 1 week overlap and extended combination groups. Both combination groups were also found to improve survival compared to abemaciclib 50 mg/kg monotherapy with 3/10 CRs.

Combination of 75 mg/kg abemaciclib, with the anti-PD-Ll antibody resulted in synergistic interaction effect for extended combo. The 1 week overlap combo was better than additive and a trended towards a significant synergistic effect. Additionally the combination of 75 mg/kg of abemaciclib with the anti-PD-Ll antibody resulted in CRs of 3/10 and 5/9 for the 1 week overlap and extended combo respectively, compared to 1/10 for 75 mg/kg abemaciclib monotherapy. Although the CR rates for the combination groups increased as compared to the monotherapy treatment at 75 mg/kg, the difference in median tumor survival at day 69 was not significant for the 1 week overlap combo, but did show a trend to significance for the extended combo.

Combination of 75 mg/kg palbociclib, with the anti-PD-Ll antibody resulted in synergistic interaction effect compared to the monotherapies in the 1 week overlap combo, and a trend towards significance synergistic effect in the extended combo.

Combination of 75 mg/kg of palbociclib with the anti-PD-Ll antibody resulted in CRs of 2/10 and 1/10 for the 1 week overlap and extended combo respectively, compared to no CRs for 75 mg/kg palbociclib monotherapy. The increase in tumor survival with the combination groups was not found to be significantly different from the monotherapy.

Also shown in Table 10 all mice that achieved CRs of their primary tumors rejected secondary rechallenge with CT26 showing that combination therapy regimens resulted in acquisition of immunologic memory which prevented the outgrowth of tumors. Mice that were partial responders at the time of rechallenge were unable to reject a secondary challenge.

Table 10: Kaplan Kaplan-Meier , Parametric Survival Analysis Of Tumor Survival For

Combination Efficacy Of Abemaciclib In CT26 Tumors

Extended combo

75 mg kg

palbociclib + anti-

34 2 2/2 <0.001 0.011 0.284 0.022 PD-L1 antibody

1 week overlap

75 mg kg

palbociclib + anti-

49.5 1 1/1 <0.001 0.003 0.137 0.078 PD-L1 antibody

Extended combo

Intra-tumor gene expression

To investigate the effects of abemaciclib treatment on the intra-tumor

microenvironment, tumor samples are prepared and processed as described in Example 2 with the following modification. In this case RNA is also analyzed for potential cell cycle inhibition resulting from monotherapy or combination treatments by analyzing for changes in the expression of various cell cycle target genes as listed in Table 11.

Table 11 - Cell Cycle Related Gene Probe List

^;HKGs, house-keeping genes

In this example, shown in Table 12, QUANTIGENE® analysis of samples from mice treated with single agent abemaciclib suggests an increase in expression of genes encoding co-stimulatory/co-inhibitory receptors and/or ligands [PD-L1 (Cd274), PD-L2 (Pdcdllg2)] and chemokines [MIPlcc (Ccl3), ΜΙΡΙβ (Ccl4), GRO (Cxcll)] that might be indicative of an induced inflammatory response. Combination of abemaciclib + anti-PD- Ll antibody resulted in marked T cell/inflamed phenotype exemplified by upregulation of 20 immune-related genes encoding leukocyte-specific [CD45 (Ptprc)], T cell-specific [CD3 (Cd3e), CD8 (Cd8bl), CD4 (Cd4)], NK cell-specific [NKG2D (Klrkl)], dendritic cell-specific [CDl lc (Itgax)], T cell activation [IFNy (Ifng)] markers, co-inhibitory receptors/ligands [PD-1/PD-L1/PD-L2 (Pdcdl, Cd274, Pdcdllg2), LAG3 (Lag3), VISTA (Vista)], immunosuppressive enzymes [IDO (Idol), iNOS (Nos2)], cytokines/chemokines [interleukin 10 (1110), TNFoc (Tnf), ΜΙΡΙα (Ccl3), ΜΙΡΙβ (Ccl4), RANTES (Ccl5), GRO (Cxcll)].

Tumors treated with anti-PD-Ll antibody monotherapy or palbociclib

monotherapy did not display a pattern of increased immune inflammatory genes.

Combination of palbociclib + anti-PD-Ll antibody produced a small signature of immune inflammation [IFNy, IDO, CD86, and PD-L2] consistent with an observable increase in combination efficacy. However, the extent of increased immune inflammation was not to the levels reached with abemaciclib combinations.

Although abemaciclib monotherapy could alter the expression of cell cycle related genes, unexpectedly, the combination of abemaciclib with an anti-PD-Ll antibody seems to enhance this effect detected in monotherapy (Table 13). In particular, treatment with 75 mg/kg of abemaciclib alone resulted in the suppression of a number of genes known to facilitate the progression of cells from the Gl phase into the S-phase or later phases of the cell cycle [ MCM7, RRM2, Topo2A, CDK2, CDK4, CCNA2, CCNB1, AURA, and MKI67]. In particular, the transcription of many of these genes is regulated by the E2F family of transcription factors whose activity is stimulated by the actions of CDK4/6. Thus the suppression of these particular genes is indicative of inhibiting CDK4/6 in the tumors. Notably increased expression of the genes for the D-cyclins (e.g. CCND2, CCND3) in the abemaciclib-treated tumors occurs concomitant with the suppression of the E2F-dependent genes. This observation is also consistent with the induction of a Gl arrest since arrest in this phase is often associated with an increase in expression of proteins such as the D-cyclins, which facilitate the progression of cells through Gl and into later phases of the cell cycle. The changes observed resulting from treatment with abemaciclib at 75 mg/kg were further enhanced during combination treatment with anti- PD-Ll therapy indicating that the combination treatments enhanced the cell cycle inhibition observed with abemaciclib alone. Similar qualitative trends in gene expression are observed during treatment with palbociclib, but the qualitative changes observed with palbociclib either alone or in combination therapy did not reach the levels observed with abemaciclib. Table 12: Combination Of Abemaciclib And Anti-PD-LI Antibody Enhances Induction Of Intra-Tumor Immune Inflammation Compared To Monotherapy And

Palbociclib Groups On Study Day 20

Two-way ANOVA, p- value < 0.05 - FC, fold change, : fold change values <1 = negative fold change, fold change values 1< = positive

Table 13: Combination Of Abemaciclib And Anti-PD-Ll Antibody Enhances

Suppression Of Intra-Tumor Cell Cycle Gene Expression Compared To Monotherapy And Palbociclib Groups

Mcm7 0.61 0.00000 Cdk2 0.70 0.00000 Edc3 0.90 0.00020

Rbll 0.73 0.00000 Cdk4 0.77 0.00000 Dnajcl4 1.08 0.00030

Ccna2 0.60 0.00010 E2f5 0.71 0.00000 Cdk4 0.88 0.00280

Ccnbl 0.61 0.00010 Mcm7 0.52 0.00000 Cdknla 0.77 0.00300

Pten 1.23 0.00020 Mki67 0.60 0.00000 Ccnbl 0.72 0.00430

Rrm2 0.60 0.00020 Rbll 0.66 0.00000 Rbll 0.85 0.00450

Aurka 0.63 0.00040 Rrm2 0.52 0.00000 Aurka 0.72 0.00630

Cdkn2c 0.58 0.00040 Aurka 0.59 0.00010 Mki67 0.78 0.02180

Mki67 0.68 0.00060 Cdkn2c 0.54 0.00010 Ccna2 0.76 0.02260

E2f5 0.82 0.00100 Top2a 0.60 0.00010 E2f4 0.92 0.04000

E2f2 0.49 0.00120 Edc3 0.91 0.00050

Dnajcl4 1.07 0.00200 Dnajcl4 1.08 0.00070

Cdk2 0.79 0.00220 Pten 1.20 0.00070

Top2a 0.69 0.00230 E2f2 0.49 0.00120

E2f4 0.90 0.01180 E2f4 0.87 0.00230

Ccndl 0.85 0.01460 Foxml 0.71 0.00490

Cdkn2a 0.62 0.01720 E2fl 0.85 0.00650

Foxml 0.76 0.02260 Ccndl 0.83 0.00710

Sdha 0.94 0.04230 Ccnel 0.75 0.01270

Cdkn2a 0.62 0.01610

Cdknla 0.83 0.03000

Sdha 0.94 0.04980

Two-way ANOVA, p-value < 0.05 - FC, fold change, : fold change values <1 = negative fold change, fold change values 1< = positive

Data presented in this example supports the following conclusions related to combination of CDK4/6 inhibitors with checkpoint immunotherapy: Pretreatment in mouse models for 1 week with CDK4/6 inhibitors such as abemaciclib or palbociclib, prior to initiation of checkpoint immunotherapy increases the response of the anti-PD-Ll antibody as compared to regimens that do not include a period with a CDK4/6 inhibitor alone. Combination therapy of CDK4/6 inhibitors with the anti-PD-Ll antibody increases the response rate and durability of CDK4/6 inhibitor therapy alone. Both one week overlap and extended combo therapies improve responses compared to

monotherapies. Increased efficacy in the one week overlap combination groups demonstrates that CDK4/6 inhibitor therapy can be discontinued during a combination treatment and still result in enhanced benefit. Increased and/or equivalent efficacy of extended combination therapy groups compared to 1 week overlap combination groups shows that CDK4/6 inhibitors can be given concurrently during the entire course of checkpoint immunotherapy after a period of CDK4/6 inhibitor monotherapy. All mice that achieved complete responses were able to resist rechallenge showing that combination therapy regimens resulted in the acquisition of immunologic memory, and the CDK4/6 inhibitor monotherapy, or CDK4/6 monotherapy prior to combination therapy does not prevent generation of immunologic memory in the combination groups. Data presented in this example supports the following conclusions related to combination of abemaciclib with checkpoint immunotherapy: Increasing the dose of abemaciclib increases the efficacy of abemaciclib combinations and durability of response. Abemaciclib combination therapy enhances immune activation in combination with an anti-PD-Ll antibody leading to improved anti-tumor activity.

Antibody expression and purification

The polypeptides of the variable regions of the heavy chain and light chain, the complete heavy chain and light chain amino acid sequences of LY3300054, and the nucleotide sequences encoding the same, are listed below in the section entitled "Amino Acid and Nucleotide Sequences." In addition, the SEQ ID NOs for the light chain, heavy chain, light chain variable region, and heavy chain variable region of LY3300054 are shown in Table 14.

The antibodies of the present invention, including, but not limited to, LY3300054 can be made and purified essentially as follows. An appropriate host cell, such as HEK 293 or CHO, can be either transiently or stably transfected with an expression system for secreting antibodies using an optimal predetermined HC:LC vector ratio or a single vector system encoding both HC and LC. Clarified media, into which the LY3300054 has been secreted, may be purified using any of many commonly-used techniques. For example, the medium may be conveniently applied to a MabSelect column (GE Healthcare), or KappaSelect column (GE Healthcare) for Fab fragment, that has been equilibrated with a compatible buffer, such as phosphate buffered saline (pH 7.4). The column may be washed to remove nonspecific binding components. The bound antibody may be eluted, for example, by pH gradient (such as 20 mM Tris buffer pH 7 to 10 mM sodium citrate buffer pH 3.0, or phosphate buffered saline pH 7.4 to 100 mM glycine buffer pH 3.0). Antibody fractions may be detected, such as by sodium dodecyl sulfate polyacrylamide gel (SDS-PAGE), and then may be pooled. Further purification is optional, depending on the intended use. The antibody may be concentrated and/or sterile filtered using common techniques. Soluble aggregate and multimers may be effectively removed by common techniques, including size exclusion, hydrophobic interaction, ion exchange, multimodal, or hydroxyapatite chromatography. The purity of the LY3300054fter these chromatography steps is greater than 95%. The product may be immediately frozen at - 70°C or may be lyophilized.

Table 14: SEQ ID NOs

Binding kinetics and affinity

The kinetics and equilibrium dissociation constant (KD) for human PD-L1 is determined for antibodies of the present invention using surface plasmon resonance (Biacore).

Immobilization of antibodies of the present invention as ligand on to sensor chip surface is performed at 25 °C. Soluble human PD-Ll-Fc fusion protein (and in some cases, cynomolgus monkey PD-Ll-Fc fusion proteins) is injected as analyte at concentrations ranging from 0.0123 nM - 9 nM. The analysis is performed at 37°C. The contact time for each sample is 180 seconds at 30 μΐνηιίηυΐε. The dissociation time was 240-1500 seconds. The immobilized surface is regenerated for 18 seconds with 0.95 M NaCl / 25 mM NaOH at 30 μυηιίηυίε, and then stabilized for 30 seconds. Binding kinetics are analyzed using the Biacore T200 Evaluation software (Version 3.0). Data are referenced to a blank flow cell, and the data are fit to a 1:1 binding model.

In experiments performed essentially as described in this assay, LY3300054 binds to human PD-L1 with a K_D of 82 pM.

Table 15: Binding By SPR Of LY3300054

ELISA analysis: LY3300054 binds to recombinant PD-L1 The ability for antibodies of the present invention to bind human PD-Ll can be measured with an ELISA assay. For the PD-Ll binding assay, a 96-well plate (NUNC®) is coated with human PD-Ll -Fc (R&D Systems) overnight at 4°C. Wells are blocked for 2 hours with blocking buffer (PBS containing 5% nonfat dry milk). Wells are washed three times with PBS containing 0.1% TWEEN®-20. Anti-PD-Ll antibody or control IgG (100 μί) is then added and incubated at room temperature for 1 hour. After washing, the plate is incubated with 100 of goat anti-human IgG F(ab')2-HRP conjugate (Jackson Immuno Research) at room temperature for 1 hour. The plates are washed and then incubated with 100 of 3,3', 5,5'-tetra-methylbenzidine. The absorbance at 450 nm is read on a microplate reader. The half maximal effective concentration (EC50) is calculated using GraphPad Prism 6 software.

In experiments performed essentially as described in this assay, LY3300054 binds to human PD-Ll with an EC50 of 0.11 nM. LY3300054 retains its binding activities after 4 weeks under all three temperature conditions, 4°C, 25°C and 40°C. LY3300054 showed a similar binding activity to PD-Ll as S70 and 2.14H90PT.

Flow cytometric analysis: LY3300054 binds to cell surface PD-Ll

The ability for antibodies of the present invention to bind to cell surface expressed human PD-Ll can be measured with a flow cytometric assay. MDA-MB 231 cells (PD- LI -positive human breast adenocarcinoma cell line) are added to a 96 well U-bottom plate at 1.5xl0⁵ cells per well in 200 μΐ, staining buffer and incubated at 4°C for 30 minutes. Plate are centrifuged at 1200 rpm for 5 minutes and supernatant removed. 100 μΐ_^ of antibody-biotin (serially diluted by 1:4 starting from 10 μg/mL) is added. A total of 6 serial dilutions are evaluated. After incubation at 4°C for 30 minutes, cells are washed twice with DPBS. 100 μΐ, of detection buffer containing 5 μΐ, streptavidin-PE is added. After incubation at 4°C for 30 more minutes, plate is centrifuged and washed twice with DPBS. Cells are re-suspended in 200 μΐ, DPBS for FACS analysis.

In experiments performed essentially as described in this assay, LY3300054 binds to cell surface PD-Ll on MDA-MB231 cells in a dose dependent manner with an EC50 of 0.14 nM. ELISA analysis: LY3300054 blocks the interaction of PD-Ll with PD-1

The ability for antibodies of the present invention to block PD-Ll binding to PD- 1 can be measured in an ELISA assay. For the receptor-ligand blocking assay, varying amounts (of anti-PD-Ll antibody or control IgG are mixed with a fixed amount of biotinylated PD-Ll -Fc fusion protein (100 ng/well) and incubated at room temperature for 1 hour. The mixture is transferred to 96- well plates pre-coated with PD-l-Fc (1 μg/mL) and then incubated at room temperature for an additional 1 hour. After washing, streptavidin HRP conjugate is added, and the absorbance at 450 nm is read. IC50 represents the LY3300054 concentration required for 50% inhibition of PD-Ll binding to PD-1.

In experiments performed essentially as described in this assay, LY3300054 blocks the interaction of PD-Ll with PD-1 with an IC50 of 0.95 nM. LY3300054 retains its blocking activities after 4 weeks under all three temperature conditions, 4°C, 25 °C and 40°C. LY3300054 demonstrates a similar ability to block PD-Ll interaction with PD-1 as S70 and 2.14H9OPT.

ELISA analysis: LY3300054 blocks the interaction of PD-Ll with B7-1

Human PD-Ll also binds to B7-1. The ability for antibodies of the present invention to block PD-Ll binding to B7-1 can be measured in an ELISA assay. The procedure for PD-L1/B7-1 blocking assay is similar to the PD-Ll/PD-1 blocking assay, except that the plates are coated with ^g/ml B7-1-Fc (R&D Systems). The antibody concentration required for 50% inhibition of PD-Ll binding to PD-1 (IC50) is calculated using GraphPad prism 6 software.

In experiments performed essentially as described in this assay, LY3300054 blocks the interaction of PD-Ll with B7-1 with an IC50 of 2.4 nM. LY3300054 shows a similar ability to block the PD-Ll interaction with the B7-1 receptor as S70 and

2.14H90PT. SEQUENCE LISTING

Amino Acid and Nucleotide Sequences

SEQ ID NO: 1 (human PD-L1)

MRIFAVFIFMTYWHLLNAFTVTVPKDLYVVEYGSNMTIECKFPVEKQLDLAALIV YWEMEDKNIIQFVHGEEDLKVQHSSYRQRARLLKDQLSLGNAALQITDVKLQDA GVYRCMISYGGADYKRITVKVNAPYNKINQRILVVDPVTSEHELTCQAEGYPKA EVIWTSSDHQVLSGKTTTTNSKREEKLFNVTSTLRINTTTNEIFYCTFRRLDPEENH TAELVIPELPLAHPPNERTHLVILGAILLCLGVALTFIFRLRKGRMMDVKKCGIQD TNSKKQSDTHLEET

SEQ ID NO: 2 (HCDR1 of LY3300054)

KASGGTFSSYAIS

SEQ ID NO: 3 (HCDR2 of LY3300054)

GIIPIFGTANYAQKFQG SEQ ID NO: 4 (HCDR3 of LY3300054)

ARSPDYSPYYYYGMDV

SEQ ID NO : 5 (LCDR 1 of LY3300054)

SGSSSNIGSNTVN

SEQ ID NO : 6 (LCDR2 of LY3300054)

YGNSNRPS

SEQ ID NO : 7 (LCDR3 of LY3300054)

QSYDSSLSGSV

SEQ ID NO: 8 (HCVR of LY3300054) QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIF GTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARSPDYSPYYYYG MDVWGQGTTVTVSS SEQ ID NO: 9 (LCVR of LY3300054)

QSVLTQPPSASGTPGQRVTISCSGSSSNIGSNTVNWYQQLPGTAPKLLIYGNSNRP SGVPDRFSGSKSGTSASLAISGLQSEDEADYYCQSYDSSLSGSVFGGGIKLTVLG

SEQ ID NO: 10 (HC of LY3300054)

QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIF GTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARSPDYSPYYYYG MDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVD KRVEPKSCDKTHTCPPCPAPEAEGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSH EDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYK CKVSNKALPSSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHE ALHNH YTQKSLS LS PGK SEQ ID NO : 11 (LC of LY3300054)

QSVLTQPPSASGTPGQRVTISCSGSSSNIGSNTVNWYQQLPGTAPKLLIYGNSNRP SGVPDRFSGSKSGTSASLAISGLQSEDEADYYCQSYDSSLSGSVFGGGIKLTVLGQ PKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPAECS

SEQ ID NO: 12 (DNA of HC of LY3300054)

CAGGTCCAGCTGGTCCAGTCAGGGGCCGAGGTCAAAAAGCCAGGGTCATCTG TCAAAGTGTCTTGTAAGGCATCCGGGGGCACATTTTCCAGCTACGCTATCTCC TGGGTGAGACAGGCACCAGGGCAGGGTCTGGAGTGGATGGGCGGAATCATTC CCATCTTCGGGACCGCCAACTACGCTCAGAAGTTTCAGGGAAGGGTCACTATT ACCGCCGACAAAAGCACATCTACTGCTTATATGGAGCTGTCTAGTCTGAGGTC TGAAGATACCGCAGTGTACTATTGCGCCCGGAGTCCCGACTATAGCCCTTACT ATTACTATGGCATGGATGTCTGGGGCCAGGGAACCACAGTGACAGTCTCATC CGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCA CCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGA ACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACC TTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGAC CGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCAC AAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACA AAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGAGGGGGCACCGTC AGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCC CTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAA GTTCAACTGGTATGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCG CGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCC TGCACCAAGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAA AGCCCTCCCATCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCC CGAGAACC AC AGGTGTAC ACCCTGCCCCCATCCCGGGAGGAGATGACCAAG A ACCAAGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCC GTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCT CCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATTCCAAGCTCACCGTGGA CAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAG GCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGCAAA

SEQ ID NO: 13 (DNA of LC of LY3300054)

CAGTCCGTCCTGACACAGCCACCCTCAGCCTCTGGCACCCCTGGGCAGCGAGT GACAATCTCTTGTTCTGGGAGTTCCTCAAATATTGGTAGTAACACCGTGAATT GGTACCAGCAGCTGCCCGGCACAGCACCTAAGCTGCTGATCTATGGAAACTC AAATAGGCCATCCGGAGTCCCCGACCGGTTCTCTGGTAGTAAATCAGGCACTT CCGCCAGCCTGGCTATTAGCGGGCTGCAGTCTGAGGACGAAGCCGATTACTA TTGCCAGTCTTACGATTCCAGCCTGTCTGGAAGTGTGTTTGGCGGAGGGATCA AGCTGACCGTCCTGGGCCAGCCTAAGGCTGCCCCCTCGGTCACTCTGTTCCCG CCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAG TGACTTCTACCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCC GTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAG TACGCGGCCAGCAGCTACCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACA GAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAG TGGCCCCTGCAGAATGCTCT SEQ ID NO: 14 (HC 178G7)

QVQLQQSGADLAKPGSSVKISCKASGYNFNSYYINWIKQTTGQGLEYIGYINTVS GTTKYSEKFKGKATLTVDKSSSTAFMQLSSLTPDDSAVYYCARGTIVLDDYWGQ GVKVTVSSAETTAPSVYPLAPGTALKSNSMVTLGCLVKGYFPEPVTVTWNSGAL SSGVHTFPAVLQSGLYTLTSSVTVPSSTWPSQTVTCNVAHPASSTKVDKKIVPRN CGGDCKPCICTGSEVSSVFIFPSKPKDVLTITLTPKVTCVVVDISQDDPEVHFSWFV DDVEVHTAQTRPPEEQFNSTFRSVSELPILHQDWLNGRTFRCKVTSAAFPSPIEKTI SKPEGRTQVPHVYTMSPTKEEMTQNEVSITCMVKGFYPPDIYVEWQMNGQPQEN YKNTPPTMDTDGSYFLYSKLNVKKEKWQQGNTFTCSVLHEGLHNHHTEKSLSHS PGK

SEQ ID NO: 15 (LC 178G7)

DIVMTQTPSSQAVSAGEKVTMSCKSSQSLLYNEKKKNYLAWYQQKPGQSPKLLI YWASTRESGVPDRFLGSGSGTDFTLTINSVQAEDLAVYYCQQSYDFPRTFGGGTK LELKRADAAPTVSIFPPSTEQLATGGASVVCLMNNFYPRDISVKWKIDGTERRDG VLDSVTDQDSKDSTYSMSSTLSLTKADYESHNLYTCEVVHKTSSSPVVKSFNRNE C

Claims

WE CLAIM:

1. A method of treating cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin's lymphoma, mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD- CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient, comprising administering to the patient an effective amount of LY3300054 and abemaciclib, or a pharmaceutically acceptable salt thereof, wherein initial administration of the abemaciclib or the salt thereof is administered to the patient without the

LY3300054 for at least 24 hours and the abemaciclib or the salt thereof is then administered to the patient in combination with the LY3300054.

2. The method according to claim 1 wherein the cancers are selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin' s lymphoma, and mantle cell lymphoma.

3. The method according to either claim 1 or 2 wherein the initial administration is for at least 1-28 days.

4. The method according to either claim 1 or 2 wherein the initial administration is for a time necessary to induce stable disease or partial response as determined by RECIST criteria or as determined by a treating physician.

5. The method according to either claim 1 or 2 wherein the initial administration is for a period of time which disease progression occurs as measured by

RECIST criteria or as determined by treating physician.

6. The method according to either claim 1 or 2 wherein the initial administration is for a time necessary to induce intra-tumor immune inflammation.

7. The method according to either claim 1 or 2 wherein the initial administration is for a time necessary to increase expression of immune inflammatory genes.

8. The method according to either claim 1 or 2 wherein the initial administration of the abemaciclib or the salt thereof to the patient is for a period of time necessary, prior to initiation of the combination, so that the subsequent combination of the abemaciclib or the salt thereof with the LY3300054 induces intra-tumor immune inflammation.

9. The method according to any one of claims 1 to 8 wherein optionally the administration of the abemaciclib or the salt thereof is stopped for a period of time determined by a treating physician to alleviate side effects of the abemaciclib or the salt thereof, and/or the LY3300054.

10. The method according to any one of claims 1 to 8 wherein optionally the administration of the abemaciclib or the salt thereof is stopped after initiation of the combination with the LY3300054, and administration of the LY3300054 is continued without administering the abemaciclib or the salt thereof.

11. The method according to any one of claims 1 to 10 wherein the abemaciclib or the salt thereof is administered orally at 150 mg approximately every 12 hours.

12. The method according to any one of claims 1 to 10 wherein the abemaciclib or the salt thereof is administered orally at 100 mg approximately every 12 hours.

13. The method according to any one of claims 1 to 12 wherein the

LY3300054 is administered at 70 mg once every two weeks by intravenous infusion.

14. The method according to any one of claims 1 to 12 wherein the

LY3300054 is administered at 200 mg once every two weeks by intravenous infusion.

15. The method according to any one of claims 1 to 12 wherein the

LY3300054 is administered at 700 mg once every two weeks by intravenous infusion.

16. The method according to any one of claims 1 to 12 wherein the

LY3300054 is administered at 1000 mg once every two weeks by intravenous infusion.

17. The method according to any one of claims 1 to 12 wherein the

LY3300054 is administered at 1200 mg once every two weeks by intravenous infusion.

18. The method according to any one of claims 1 to 12 wherein the

LY3300054 is administered at 1400 mg once every two weeks by intravenous infusion.

19. The method according to any one of claims 1 to 12 wherein the

LY3300054 is administered at a dose level needed to achieve constant saturation of available PD-Ll target protein by intravenous infusion during the course of combination therapy.

20. The method according to any one of claims 1 to 19 wherein the abemaciclib or the salt thereof is administered alone for 2 weeks of a first 28-day cycle and then starting on Day 15 of the first 28-day cycle and thereafter on Days 1 and 15 of subsequent cycles, the abemaciclib or the salt thereof is administered in combination with the LY3300054.

21. The method according to claim 20 wherein on days the abemaciclib and the LY3300054 are administered on the same day, the abemaciclib or the salt thereof is administered approximately 2 hours after starting the administration of the LY3300054.

22. A combination comprising LY3300054 and abemaciclib, or a

pharmaceutically acceptable salt thereof, for simultaneous, separate, or sequential use in the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin' s lymphoma, mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6- Rb-E2F pathway), and additional susceptible tumors in a patient wherein initial administration of the abemaciclib or the salt thereof is administered to the patient without the LY3300054 for at least 24 hours and the abemaciclib or the salt thereof is then administered to the patient in simultaneous, separate, or sequential combination with the LY3300054.

23. LY3300054 for use in simultaneous, separate, or sequential combination with a compound which is abemaciclib, or a pharmaceutically acceptable salt thereof, in the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin' s lymphoma, mantle cell lymphoma, additional tumors which display alterations in cell cycle control (CyclinD- CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein the initial administration of the abemaciclib or the salt thereof is administered to the patient without the LY3300054 for at least 24 hours and the abemaciclib or the salt thereof is then administered to the patient in simultaneous, separate, or sequential combination with the LY3300054.

24. Abemaciclib, or a pharmaceutically acceptable salt thereof, for use in simultaneous, separate, or sequential combination with LY3300054 in the treatment of a cancer selected from the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin' s lymphoma, mantle cell lymphoma, tumors which display alterations in cell cycle control (CyclinD-CDK4/CDK6-Rb-E2F pathway), and additional susceptible tumors in a patient wherein the initial administration of the abemaciclib or the salt thereof is administered without the LY3300054 for at least 24 hours and the abemaciclib or the salt thereof is then administered in simultaneous, separate, or sequential combination with the LY3300054.

25. The combination, the LY3300054, or the abemaciclib or the salt thereof, for use according to any one of claims 22 to 24 wherein the cancer is selected form the group consisting of NSCLC, breast cancer, melanoma, colon cancer, pancreatic cancer, non-Hodgkin' s lymphoma, and mantle cell lymphoma.

26. The combination, the LY3300054, or the abemaciclib or the salt thereof, for use according to any one of claims 22 to 25 wherein the initial administration is for at least 1-28 days.

27. The combination, the LY3300054, or the abemaciclib or the salt thereof, for use according to any one of claims 22 to 25 wherein the initial administration is for a time necessary to induce stable disease or partial response as determined by RECIST criteria or as determined by a treating physician.

28. The combination, the LY3300054, or the abemaciclib or the salt thereof, for use according to any one of claims 22 to 25 wherein the initial administration is for a period of time which disease progression occurs as measured by RECIST criteria or as determined by treating physician.

29. The combination, the LY3300054, or the abemaciclib or the salt thereof, for use according to any one of claims 22 to 25 wherein the initial administration is for a time necessary to induce intra-tumor immune inflammation.

30. The combination, the LY3300054, or the abemaciclib or the salt thereof, for use according to any one of claims 22 to 25 wherein the initial administration is for a time necessary to increase expression of immune inflammatory genes.

31. The combination, the LY3300054, or the abemaciclib or the salt thereof, for use according to any one of claims 22 to 25 wherein the initial administration of the abemaciclib or the salt thereof to the patient is for a period of time necessary, prior to initiation of the combination, that the subsequent combination of the abemaciclib or the salt thereof with the LY3300054 induces intra-tumor immune inflammation.

32. The combination, the LY3300054, or the abemaciclib or the salt thereof, for use according to any one of claims 22 to 31 wherein the abemaciclib or the salt thereof is administered orally at 150 mg approximately every 12 hours.

33. The combination, the LY3300054, or the abemaciclib or the salt thereof, for use according to any one of claims 22 to 31 wherein the abemaciclib or the salt thereof is administered orally at 100 mg approximately every 12 hours.

34. The combination, the LY3300054, or the abemaciclib or the salt thereof, for use according to any one of claims 22 to 33 wherein the LY3300054 is administered at 70 mg once every two weeks by intravenous infusion.

35. The combination, the LY3300054, or the abemaciclib or the salt thereof, for use according to any one of claims 22 to 33 wherein the LY3300054 is administered at 200 mg once every two weeks by intravenous infusion.

36. The combination, the LY3300054, or the abemaciclib or the salt thereof, for use according to any one of claims 22 to 33 wherein the LY3300054 is administered at 700 mg once every two weeks by intravenous infusion.

37. The combination, the LY3300054, or the abemaciclib or the salt thereof, for use according to any one of claims 22 to 33 wherein the LY3300054 is administered at 1000 mg once every two weeks by intravenous infusion.

38. The combination, the LY3300054, or the abemaciclib or the salt thereof, for use according to any one of claims 22 to 33 wherein the LY3300054 is administered at 1200 mg once every two weeks by intravenous infusion.

39. The combination, the LY3300054, or the abemaciclib or the salt thereof, for use according to any one of claims 22 to 33 wherein the LY3300054 is administered at 1400 mg once every two weeks by intravenous infusion.

40. The combination, the LY3300054, or the abemaciclib or the salt thereof, for use according to any one of claims 22 to 33 wherein the LY3300054 is administered at a dose level needed to achieve constant saturation of available PD-Ll target protein by intravenous infusion during the course of combination therapy. .

41. The combination, the LY3300054, or the abemaciclib or the salt thereof, for use according to any one of claims 22 to 40 wherein the abemaciclib or the salt thereof is administered alone for 2 weeks of a first 28-day cycle and then starting on Day 15 of the first 28-day cycle and thereafter on Days 1 and 15 of subsequent cycles, the abemaciclib or the salt thereof is administered in combination with the LY3300054.

42. The combination, the LY3300054, or the abemaciclib or the salt thereof, for use according to claim 41 wherein on days the abemaciclib and the LY3300054 are administered on the same day, the abemaciclib or the salt thereof is administered approximately 2 hours after starting the administration of the LY3300054