Therapeutic compositions, combinations, and methods of use

Abstract

This invention relates to methods comprising administering a FAK inhibitor and an immunotherapeutic agent such as anti-PD-1 or anti-PD-L1; that are useful in the treatment of abnormal cell growth, such as cancer, in mammals, especially humans.

Claims

1. A method for treating a human subject suffering from cancer, comprising administering an effective amount of a FAK inhibitor selected from VS-6063 and VS-4718, or a pharmaceutically acceptable salt thereof, in combination with an anti-PD-1 antibody, wherein the cancer is selected from the group consisting of mesothelioma, neurofibromatosis, renal cancer, lung cancer, non-small cell lung cancer, liver cancer, thyroid cancer, ovarian cancer, breast cancer, schwannoma, meningioma, schwannomatosis, acoustic neuroma, adenoid cystic carcinoma, ependymoma, and ependymal tumors.

2. The method of claim 1, wherein the cancer is solid tumor.

3. The method of claim 1, wherein the FAK inhibitor is administered orally.

4. The method of claim 3, wherein the FAK inhibitor is administered at about 100 mg to about 2000 mg.

5. The method of claim 3, wherein the FAK inhibitor is VS-6063 and the FAK inhibitor is administered at about 200 mg to about 600 mg twice a day.

6. The method of claim 3, wherein the FAK inhibitor is VS-4718 and the FAK inhibitor is administered at about 300 mg to about 500 mg once a day.

7. The method of claim 6, wherein the FAK inhibitor is VS-4718 and the FAK inhibitor is administered at about 200 mg to about 400 mg twice a day.

8. The method of claim 1, wherein the anti-PD-1 antibody is administered parenterally.

9. The method of claim 8, wherein the anti-PD-1 antibody is selected from the group consisting of: nivolumab, AMP-224, pembrolizumab, and pidilizumab.

10. The method of claim 1, wherein the subject has been previously treated with a chemotherapeutic agent or with radiation therapy.

11. The method of claim 1, wherein the subject has failed conventional or standard cancer treatment.

12. The method of claim 1, wherein the subject has failed first-line treatment.

13. The method of claim 1, wherein the anti-PD-1 antibody is nivolumab.

14. The method of claim 1, wherein the anti-PD-1 antibody is pembrolizumab.

15. The method of claim 1, wherein the anti-PD-1 antibody is AMP-224.

16. The method of claim 1, wherein the anti-PD-1 antibody is pidilizumab.

17. The method of claim 1, wherein the cancer is non-small cell lung cancer.

18. The method of claim 1, wherein the cancer is ovarian cancer.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIGS. 1A-1B show an exemplary effect of FAK Inhibitors as compared to other Tyrosine Kinase

(2) Inhibitors on T-cell proliferation.

(3) FIG. 2 shows an exemplary effect of FAK Inhibitor VS-4718 alone and in combination with anti-PD-1 on the survival of a colorectal cancer model.

(4) FIGS. 3A-3D show an exemplary effect of blockers of the immune checkpoint pathway and correlation with biomarkers of the immune response.

(5) FIG. 4 shows an exemplary enhancement of anti-tumor efficacy of co-stimulatory antibodies by FAK inhibitors.

(6) FIG. 5 shows an exemplary effect of FAK inhibitors on CD8+ T cells.

(7) FIGS. 6A-6B show exemplary effect of FAK inhibitors on T cell exhaustion markers.

EXAMPLES

(8) The disclosure is further described in the following examples, which do not limit the scope of the claims.

Example 1. FAK/PYK2 Inhibition Enhances Efficacy of Immune Checkpoint Inhibition

(9) The small molecule FAK/PYK2 inhibitor VS-6063 is shown to inhibit monocyte-derived macrophages, decreases IL-6 and IL-8 production from macrophages in vitro, and reduces tumor-associated macrophages in xenograft models. Additionally, in contrast to other protein kinase inhibitors, such as the SRC inhibitor dasatinib and the MEK inhibitor trametinib which potently impair the proliferation of CD8+ cytotoxic T-cells, VS-4718 and VS-6063 stimulate proliferation of CD8+ cytotoxic T-cells (FIG. 1). Primary human CD8+ T-cells isolated from healthy donor PBMCs were incubated in the presence of anti-CD.sup.3/anti-CD28 coated beads with increasing concentrations of VS-4718 or VS-6063 for 72-hours and assayed for BrdU incorporation as a measure of new DNA synthesis. Both FAK inhibitors dose-dependently increased CD8+ T-cell proliferation.

(10) Based on the inhibition of tumor-associated macrophages and enhancement of CD8+ T-cells, potentiation of FAK/PYK2 inhibitors on the anti-tumor efficacy of an anti-PD-1 monoclonal antibody in syngeneic mouse tumor models was investigated. Mice bearing established syngeneic MC38 colorectal tumors were treated with VS-4718 for 5 days before combination treatment with anti-PD1 antibody along with continued VS-4718 administration. Combination of VS-4718 with anti-PD1 extended the median overall survival to 42 days relative to 21, 25 and 28 day median overall survival with vehicle control, single agent anti-PD-1 and single agent VS-4718, respectively (FIG. 2). Moreover, on day 56, 30% of mice treated with the VS-4718/anti-PD-1 combination were still surviving compared to no surviving mice in the vehicle, single agent VS-4718 and single agent anti-PD-1 groups.

(11) FAK kinase inhibitor or FAK genetic ablation each induced full tumor regression in a Squamous cell carcinoma model through an immune mechanism, suggested by an increase in CD8+ and CD4+ T-cells, and a decrease in T-regs.

(12) The general pattern of immune cell changes in response to FAK inhibitors emerged across syngeneic models of skin, pancreatic, lung, and breast cancers. Increased cytotoxic T-cells were observed in tumor (for example, the CD8+ T-cell population). A decrease in immune cell populations that suppress anti-tumor immune response (for example, T-regs, M2 tumor-associated macrophages, MDSCs) was also observed. A pattern of an increase in PDL1-High tumor cells and an increase in PD-1 and cytotoxic T-cells was also seen. The data suggests synergy between FAK inhibitors and immune checkpoint antibodies (anti-PD-1 anti-CTLA-4) in multiple tumor models.

Example 2. Efficacy of FAK Inhibitors with Anti-PD-1 and Correlation with Biomarkers

(13) Mice bearing syngeneic MC38 tumors were randomized once tumors reached 50-100 mm.sup.3 and treated with either vehicle, VS-4718 (75 mg/kg, BID, p.o. through end of experiment), anti-PD1 (clone RMP1-14, 10 mg/kg i.p. on days 1, 4, 8, 11) or VS-4718+anti-PD1. Median tumor volume over the days elapsed is shown in FIG. 3A. The combination of VS-4718 and anti-PD1 improved anti-tumor efficacy in syngeneic MC38 colorectal tumor-bearing mice.

(14) Syngeneic MC38 tumors were randomized once tumors reached 50-100 mm.sup.3 and then treated with either vehicle, VS-4718 (50 mg/kg, BID), anti-PD1 (clone RMP1-14, 5 mg/kg on days 1, 4, 8, 11) or VS-4718+anti-PD1 for 12 days at which point tumors were processed live for flow cytometry (FIG. 3B-D). VS-4718 and anti-PD1 combination in MC38 tumors decreased Tregs and increased CD8+ T cells.

(15) Syngeneic MC38 tumors were randomized once tumors reached 50-100 mm.sup.3 and then treated with either vehicle, VS-4718 (75 mg/kg, BID, po), anti-4-1BB (clone LOB12.3, 10 mg/kg i.p. on days 1, 4, 8, 11) or VS-4718+anti-4-1BB. Mean tumor volume is plotted over time in FIG. 4. The combination of VS-4718 and anti-4-1BB improves anti-tumor efficacy in syngeneic MC38 colorectal tumor-bearing mice.

(16) CD8+ T cells were isolated from fresh, healthy human PBMCs by negative immunomagnetic bead separation. Purified CD8+ T cells were plated on CD3-coated plates in the presence or absence of VS-4718, VS-6063, or GSK2256098 for 72 hours. Assay wells were pulsed with BrdU for the last 3-4 hours of culture and subjected to a BrdU-incorporation assay for the determination of actively proliferating cells. Data shown in FIG. 5 are presented as fold-change vs. DMSO control wells.

(17) CD8+ T cells isolated from fresh, healthy human PBMCs by negative immunomagnetic bead separation were plated on anti-CD3 coated plates in the presence of VS-4718 (FIG. 6A) or VS-6063 (FIG. 6B) for 72 hours and then harvested and stained with anti-LAG3 or anti-PD-1 for flow cytometric analysis.

(18) FAK inhibitors change the tumor immune balance to potentiate efficacy of various immuno-oncology agents. FAK inhibitor combination substantially enhances anti-tumor efficacy of anti-PD-1 or anti-4-1BB vs. each immuno-oncology antibody alone. FAK inhibitor+anti-PD-1 combination decreases Tregs and increases CD8+ T cells in MC38 tumors. FAK inhibitors increase CD8+ T cell proliferation, decrease CD8+ T cell exhaustion markers, and increase T cell-mediated tumor cell killing in vitro.