Pharmaceutical combination

Abstract

To provide a pharmaceutical combination for treating diseases and / or disorders related to the Wnt pathway.SOLUTION: The present invention provides a pharmaceutical combination comprising a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof and an anti-PD-1 antibody or a pharmaceutically acceptable salt or solvate thereof.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates generally to the field of oncology. In particular, the present invention relates to a pharmaceutical combination comprising a compound of formula (I) and an anti-PD-1 antibody for treating diseases and / or disorders associated with the Wnt pathway. [Background technology]

[0002] Reference in this specification to any prior publication (or information derived therefrom) or to any known matter is not, and should not be, taken as an admission or acknowledgment, or any form of suggestion, that the prior publication (or information derived therefrom) or known matter forms part of the common general knowledge in the field to which this specification pertains.

[0003] Cancers are typically treated with chemotherapy and / or radiation therapy. Although such therapies are often effective in destroying a significant amount of tumor cells, they often leave some tumor cells resistant to the treatment. These resistant cells can proliferate and thus form new tumors that are resistant to the treatment. As a result, the regular use of known combinations of chemotherapy drugs has given rise to multidrug resistant ("MDR") tumor cells.

[0004] The mechanism of proliferative diseases such as tumors is multifactorial. For example, research over the last 40 years has led to the recognition that cytotoxic (or antiproliferative) agents include antimetabolites that interfere with microtubule assembly, alkylating agents that can cross-link DNA, platinum agents that can interfere with DNA by blocking DNA replication, antitumor antibiotics, topoisomerase inhibitors, etc. In the treatment of such diseases, drugs with different mechanisms may be combined with beneficial effects, including effective treatment of MDR tumor cells, and may also be combined to minimize undesirable side effects such as cytotoxicity (i.e., combination therapy). The problem here is that not all known antiproliferative agents provide useful or beneficial effects in combination, and therefore research in many laboratories is currently focused on developing new useful antiproliferative combination partners.

[0005] It is therefore generally desirable to provide a pharmaceutical combination that overcomes or ameliorates one or more of the above mentioned problems. Summary of the Invention

[0006] The present invention relates to a) Formula (I) having the following structure:

[0007] [ka]

[0008] or a pharma- ceutically acceptable salt or solvate thereof, b) an anti-PD-1 antibody molecule, or a pharma- ceutically acceptable salt or solvate thereof; The present invention provides a pharmaceutical combination comprising:

[0009] In one embodiment, the anti-PD-1 antibody molecule is pembrolizumab. Disclosed herein is a method of treating a proliferative disease or disorder associated with the Wnt pathway in a subject, the method comprising administering to the subject a pharmaceutical combination defined herein.

[0010] In certain embodiments, the proliferative disease or condition is colorectal cancer or endometrial cancer. Disclosed herein is a pharmaceutical combination as defined herein for use in the treatment of a proliferative disease or disorder associated with the Wnt pathway in a subject.

[0011] Disclosed herein is the use of a pharmaceutical combination as defined herein in the manufacture of a medicament for the treatment of a proliferative disease or disorder associated with the Wnt pathway in a subject.

[0012] Aspects of the present invention will now be described, by way of non-limiting example only, with reference to the accompanying drawings, in which: [Brief description of the drawings]

[0013] [Figure 1-1] FFPE tumor tissue from patient 202-101 with a tumor of unknown primary site (presumed cholangiocarcinoma) was prepared from paired biopsies pre- and post-C1D17 treatment. Sections were assayed via IHC / OPAL and analyzed on a Vectra3 (Perkin Elmer). Percentages of CD8+ vs. FOXP3+ cells and CD4+ vs. FOXP3+ cells are plotted using counts from three high power fields measured on single stained sections. [Figure 1-2] FFPE tumor tissue from patient 202-101 with a tumor of unknown primary site (presumed cholangiocarcinoma) was prepared from paired biopsies pre- and post-C1D17 treatment. Sections were assayed via IHC / OPAL and analyzed on a Vectra3 (Perkin Elmer). Percentages of CD8+ vs. FOXP3+ cells and CD4+ vs. FOXP3+ cells are plotted using counts from three high power fields measured on single stained sections. [Figure 2-1] Nanostring analysis of FFPE and frozen tumor tissues from patient 202-101 (T4-T3) with tumor of unknown primary site (probable cholangiocarcinoma) and MSS CRC102-102 (T2-T1) patient (with RSPO gene fusion) using a Pan-Cancer immune panel. (A) Grouped analysis showing similar pre- and post-treatment changes in both samples. (B) Most up- and down-regulated genes common to both cancer types. (C and D) Most up- and down-regulated genes in CRC patient with RSPO fusion and tumor of unknown primary site (presumed cholangiocarcinoma), respectively. Red gene symbols indicate genes up- or down-regulated in both patients. [Figure 2-2] Nanostring analysis of FFPE and frozen tumor tissues from patient 202-101 (T4-T3) with tumor of unknown primary site (probable cholangiocarcinoma) and MSS CRC102-102 (T2-T1) patient (with RSPO gene fusion) using a Pan-Cancer immune panel. (A) Grouped analysis showing similar pre- and post-treatment changes in both samples. (B) Most up- and down-regulated genes common to both cancer types. (C and D) Most up- and down-regulated genes in CRC patient with RSPO fusion and tumor of unknown primary site (presumed cholangiocarcinoma), respectively. Red gene symbols indicate genes up- or down-regulated in both patients. [Figure 2-3]Nanostring analysis of FFPE and frozen tumor tissues from patient 202-101 (T4-T3) with tumor of unknown primary site (probable cholangiocarcinoma) and MSS CRC102-102 (T2-T1) patient (with RSPO gene fusion) using a Pan-Cancer immune panel. (A) Grouped analysis showing similar pre- and post-treatment changes in both samples. (B) Most up- and down-regulated genes common to both cancer types. (C and D) Most up- and down-regulated genes in CRC patient with RSPO fusion and tumor of unknown primary site (presumed cholangiocarcinoma), respectively. Red gene symbols indicate genes up- or down-regulated in both patients. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0014] The present invention relates to a) Formula (I) having the following structure:

[0015] [ka]

[0016] or a pharma- ceutically acceptable salt or solvate thereof, b) an anti-PD-1 antibody molecule, or a pharma- ceutically acceptable salt or solvate thereof; The present invention provides a pharmaceutical combination comprising:

[0017] Without wishing to be bound by theory, the present invention is based on the discovery that the compound of formula (I) is compatible with anti-PD-1 antibody molecules as a combination therapy for treating cancer, such as microsatellite stable (MSS) colorectal cancer or endometrial cancer. The inventors have found that the compound of formula (I) (Compound A) leads to an increase in infiltrating lymphocytes in tumors and inhibits FOXP 3+ CD8 for T cells +They found that the anti-PD-1 antibody increased the proportion of T cells in the tumor. In addition, nanostring analysis showed that genes such as PDCD1 and IFNG were highly upregulated, while other genes such as the checkpoint kinase ATM and the chemokine CCL15 were highly downregulated. The increase in tumor-infiltrating lymphocytes (TILs) and the change in the tumor environment suggest that the tumor has changed from an immunologically "cold" tumor to an immunologically "hot" tumor, which can be further treated with anti-PD-1 antibody molecules.

[0018] In the present disclosure, the term "pharmaceutical combination" refers to a non-fixed combination. The term "non-fixed combination" refers to a combination of active ingredients, such as a compound of formula (i), i.e., formula (I) having the following structure:

[0019] [ka]

[0020] or a pharma- ceutically acceptable salt or solvate thereof and the anti-PD-1 antibody molecule, or a pharma- ceutically acceptable salt or solvate thereof, are administered together to a patient, either simultaneously as separate entities, or sequentially without any specific time limit, such administration resulting in therapeutically effective levels of the two compounds in the body of said patient.

[0021] The term "combination" or "in combination with" is not intended to imply that the therapies or therapeutic agents must be administered simultaneously and / or formulated for delivery together, although these delivery methods are within the scope described herein. The therapeutic agents in the combination may be administered simultaneously with, prior to, or subsequent to one or more other additional therapies or therapeutic agents. The therapeutic agents or treatment protocols may be administered in any order. Generally, each agent will be administered at a dose and / or time schedule determined for that agent. It will be further understood that the additional therapeutic agents used in the combination may be administered together as different compositions or separately. In general, it is expected that the additional therapeutic agents used in the combination will be used at levels that do not exceed the levels at which they are used individually. In some embodiments, the levels used in combination will be lower than the levels used individually.

[0022] For example, the anti-PD-1 antibody molecule may be administered prior to administration of the compound of Formula (I). The anti-PD-1 antibody molecule may be administered after administration of the compound of Formula (I). The compound of Formula (I) may be administered simultaneously with the anti-PD-1 antibody molecule. In one embodiment, the compound of Formula (I) is administered once daily (e.g., orally) while the anti-PD-1 antibody molecule is administered once every 21 days (e.g., by intravenous infusion).

[0023] The combination partners described herein may be synergistically active while reducing side effects caused by Wnt signaling pathway inhibition, such as reduced bone mineral density. The term "pharmaceutically acceptable salt" may be formed, for example, as an acid addition salt, preferably with organic or inorganic acids. Suitable inorganic acids are, for example, halogen acids, such as hydrochloric acid. Suitable organic acids are, for example, carboxylic or sulfonic acids, such as fumaric or methanesulfonic acid. For isolation or purification purposes, it is also possible to use pharmaceutically unacceptable salts, for example, picrates or perchlorates. For therapeutic use, only pharmaceutically acceptable salts or free compounds are employed (applicable in the form of pharmaceutical preparations), and therefore, these are preferred. All references to free compounds herein should be understood to also refer to the corresponding salts, where necessary and convenient. The salts of the inhibitors described herein are preferably pharmaceutically acceptable salts, and suitable counterions forming pharmaceutically acceptable salts are known in the art.

[0024] The term "pharmaceutical acceptable" means suitable for use in contact with the tissues of human and animals without undue toxicity, irritation, allergic response, or other problem or complication. It refers to compounds, substances, compositions, and / or dosage forms that are associated with a reasonable benefit / risk ratio.

[0025] The term "inhibition" or "inhibitor" includes a decrease in a particular parameter, e.g., activity, of a given molecule, e.g., an immune checkpoint inhibitor, such as an anti-PD-1 antibody molecule. For example, at least 5%, 10%, 20%, 30%, 40% or more inhibition of activity, e.g., PD-1 or PD-L1 activity, is encompassed by the term. Thus, inhibition need not be 100%.

[0026] The term "antibody molecule" refers to a protein that comprises at least one immunoglobulin variable domain sequence, e.g., an immunoglobulin chain or fragment thereof. The term "antibody molecule" includes, for example, monoclonal antibodies (including full-length antibodies having an immunoglobulin Fc region). An antibody molecule includes a full-length antibody, a full-length immunoglobulin chain, or an antigen-binding or functional fragment of a full-length antibody or full-length immunoglobulin chain. An antibody molecule can also be a multispecific antibody molecule, e.g., comprising a plurality of immunoglobulin variable domain sequences, a first immunoglobulin variable domain sequence of the plurality having binding specificity for a first epitope and a second immunoglobulin variable domain sequence of the plurality having binding specificity for a second epitope.

[0027] The antibody molecule may be derivatized or conjugated to another functional molecule (e.g., another peptide or protein). Methods of derivatization include, but are not limited to, the addition of a fluorescent moiety, a radioactive nucleotide, a toxin, an enzyme, or an affinity ligand such as biotin. Thus, the antibody molecule of the present invention is intended to include derivatized and otherwise modified forms of the antibodies described herein, including immunoadhesion molecules. For example, the antibody molecule may be functionally conjugated (by chemical coupling, genetic fusion, non-covalent association, or otherwise) to one or more other molecular entities, such as another antibody (e.g., a bispecific antibody or diabody), a detectable agent, a cytotoxic agent, a pharmaceutical agent, and / or a protein or peptide that can mediate the association of the antibody or antibody portion with another molecule (such as a streptavidin core region or a polyhistidine tag).

[0028] By "binding fragment" or "antigen-binding fragment" of an antibody, it is intended to refer to any peptide, polypeptide, or protein that retains the ability to bind to the antibody's target (also commonly referred to as the antigen). In certain embodiments, such an "antigen-binding fragment" is selected in the group consisting of Fv, scFv (sc is single chain), Fab, F(ab')2, Fab', scFv-Fc fragments or diabodies, or any fragment whose half-life will be extended by chemical modification such as the addition of poly(alkylene) glycols such as poly(ethylene)glycol ("pegylation") (pegylated fragments referred to as Fv-PEG, scFv-PEG, Fab-PEG, F(ab')2-PEG, or Fab'-PEG) ("PEG" is poly(ethylene)glycol), or by incorporation into liposomes. In one embodiment, an "antigen-binding fragment" will be constituted by or comprise a subsequence of the variable heavy or variable light chain of the antibody from which it is derived sufficient to retain the same binding specificity for a target as the antibody from which it is derived and sufficient affinity, for example at least equal to 1 / 100, and as a further example at least equal to 1 / 10, of the affinity of the antibody from which it is derived.

[0029] The anti-PD-1 antibody molecule may be an anti-PD-1 antibody known in the art. In one embodiment, the anti-PD-1 antibody is MDX-1106 (also known as MDX-1106-04, ONO-4538, BMS-936558, or nivolumab). In one embodiment, the anti-PD-1 antibody is nivolumab (CAS Registry Number: 946414-94-4). Nivolumab (also known as BMS-936558 or MDX1106) ;Bristol-Myers Squibb) is a fully human IgG4 monoclonal antibody that specifically blocks PD-1. Nivolumab (clone 5C4) and other human monoclonal antibodies that specifically bind to PD-1 are disclosed in U.S. Patent No. 8,008,449 and WO2006 / 121168. Lambrolizumab (also called pembrolizumab or MK03475; Merk) is a humanized IgG4 monoclonal antibody that specifically binds to PD-1. Pembrolizumab is disclosed in U.S. Patent No. 8,354,509 and WO2009 / 114335. In some embodiments, the anti-PD-1 antibody is pidilizumab. Pidilizumab (CT-011; Cure Tech) is a humanized IgGlk monoclonal antibody that binds to PD1. Pidilizumab and other humanized anti-PD-1 monoclonal antibodies are disclosed in WO2009 / 101611. In one embodiment, the anti-PD-1 antibody is PDR-001, a humanized monoclonal antibody described in WO2015 / 112900. Other anti-PD1 antibodies are disclosed in U.S. Patent No. 8,609,089, U.S. Patent Application Publication No. 2010028330, and / or U.S. Patent Application Publication No. 20120114649. Other anti-PD-1 antibodies include AMP514 (Amplimmune).

[0030] The anti-PD-1 antibody molecule may be selected from nivolumab (Opdivo), pembrolizumab (Keytruda), pidilizumab, or PDR-001. In one embodiment, the anti-PD-1 antibody molecule is pembrolizumab.

[0031] It is understood that the anti-PD-1 antibody molecule, i.e., the anti-PD-1 antibody molecule of the present disclosure, may have additional conservative or non-essential amino acid substitutions that do not significantly affect its function.

[0032] Disclosed herein is a method of treating a proliferative disease or disorder associated with the Wnt pathway in a subject, comprising administering to said subject a pharmaceutical combination as defined herein. The pharmaceutical combination can be administered for a time and under conditions sufficient to treat the proliferative disease or disorder associated with the Wnt pathway in the subject.

[0033] The term "treatment" includes, for example, the therapeutic administration of a combination of a Wnt inhibitor or a pharma- ceutically acceptable salt thereof and an anti-PD-1 antibody molecule or a pharma- ceutically acceptable salt thereof as described herein to a warm-blooded animal, particularly a human, in need of such treatment, with a view to curing said disease or to effect regression of disease or delay of disease progression. The terms "treat", "treating" or "treatment" of any disease or disorder refer to ameliorating said disease or disorder (e.g., delaying or inhibiting or alleviating the onset of said disease or at least one of its clinical symptoms), preventing or delaying the onset or onset or progression of said disease or disorder.

[0034] The term "patient" or "subject" refers to a warm-blooded animal. In the most preferred embodiment, the subject or patient is a human. This may be a human who has been diagnosed and is in need of a treatment as disclosed herein for a disease or disorder.

[0035] The method defined herein may include administering an effective amount of the pharmaceutical combination to a subject in need thereof. The term "effective amount" as defined herein means administering an amount of the drug to an individual in need thereof, either in a single dose or as part of a series that is effective for the induction, treatment, or prevention thereof. The effective amount will vary according to the health and physical condition of the individual being treated, the taxonomic group of the individual being treated, the formulation of the combination, the evaluation of the medical condition, and other relevant factors. It is expected that the amount will fall within a relatively broad range that can be determined through routine trials.

[0036] The terms "cancer" and "cancerous" refer to or refer to a physiological condition in a mammal that is typically characterized in part by uncontrolled cell proliferation. As used herein, the term "cancer" refers to non-metastatic and metastatic cancers, including early and late cancers. The term "pre-cancerous" refers to a condition or growth that typically precedes or progresses to cancer. "Non-metastatic" refers to a cancer that is benign or remains at the primary site and has not invaded the lymphatic or vascular system or tissues other than the primary site. Generally, a non-metastatic cancer is any cancer that is a stage 0, I, or II cancer, and sometimes a stage III cancer. "Early cancer" refers to a cancer that is not invasive or metastatic or is classified as a stage 0, I, or II cancer. The term "late cancer" generally refers to stage III or stage IV cancer, but can also refer to stage II cancer or a sub-stage of stage II cancer. Those skilled in the art will appreciate that the classification of stage II cancer as either early or late cancer depends on the particular type of cancer. Illustrative examples of cancer include, but are not limited to, breast cancer, prostate cancer, ovarian cancer, cervical cancer, pancreatic cancer, colorectal cancer, lung cancer, hepatocellular carcinoma, gastric cancer, liver cancer, bladder cancer, cancer of the urinary tract, thyroid cancer, renal cancer, carcinoma, melanoma, brain cancer, non-small cell lung cancer, squamous cell carcinoma of the head and neck, uterine cancer, multiple myeloma, rectal cancer, and esophageal cancer.

[0037] In certain embodiments, the proliferative disease or disorder is associated with upregulation of the Wnt pathway. In one embodiment, the proliferative disease or condition is selected from the group consisting of cancer, fibrosis, stem cell and diabetic retinopathy, rheumatoid arthritis, psoriasis, and myocardial infarction.

[0038] In one embodiment, the proliferative disease or condition is selected from the group consisting of cervical cancer, colon cancer, colorectal cancer, breast cancer, bladder cancer, uterine cancer, head and neck cancer, gastric cancer, lung cancer, ovarian cancer, prostate cancer, thyroid cancer, non-small cell lung cancer, chronic lymphocytic leukemia, mesothelioma, melanoma, pancreatic adenocarcinoma, basal cell carcinoma, osteosarcoma, hepatocellular carcinoma, Wilms' tumor, medulloblastoma, pulmonary fibrosis, liver fibrosis, skin fibrosis, renal fibrosis, stem cell and diabetic retinopathy, rheumatoid arthritis, psoriasis, and myocardial infarction.

[0039] In one embodiment, the proliferative disease or condition is lung cancer, prostate cancer, melanoma, osteosarcoma, colorectal cancer, or endometrial cancer. In one embodiment, the proliferative disease or condition is colorectal cancer.In one embodiment, the proliferative disease or condition is microsatellite-stable colorectal cancer.

[0040] In one embodiment, the proliferative disease or condition is endometrial cancer. The administration of the compound of formula (I) can vary and can be at intervals of minutes, hours, days, weeks, months, or years, or continuously over any one of these time periods. In one embodiment, the method comprises administering the compound of formula (I) to the subject once every day. In one embodiment, the method comprises administering the compound of formula (I) once every two days (i.e., every other day). In one embodiment, the method comprises administering the compound of formula (I) once every third, fourth, fifth, sixth, or seventh day. In one embodiment, the method comprises administering the compound of formula (I) two or three times per day. The method may comprise orally administering the compound of formula (I) to the subject. The compound of formula (I) may be administered in a single dose or in a series of doses. The compound of formula (I) may be administered as individual units or a combination of individual units.

[0041] The administration schedule of the anti-PD-1 antibody molecule may be, for example, once a week for two weeks to three weeks. The period may vary from once a week, to once every four weeks. In one embodiment, the method comprises administering an anti-PD-1 antibody molecule to the subject once every 21 days. The method may comprise administering an anti-PD-1 antibody molecule to the subject by intravenous infusion. The method may comprise administering an anti-PD-1 antibody molecule to the subject by intravenous injection.

[0042] The compound of formula (I) is administered in an amount of about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 9 mg, about 10 mg, about 11 mg, about 12 mg, about 13 mg, about 14 mg, about 15 mg, about 16 mg, about 17 mg, about 18 mg, about 19 mg, about 20 mg, about 21 mg, about 22 mg, about 23 mg, about 24 mg, about 25 mg, about 26 mg, about 27 mg, about 28 mg , about 29 mg, about 30 mg, about 31 mg, about 32 mg, about 33 mg, about 34 mg, about 35 mg, about 36 mg, about 37 mg, about 38 mg, about 38 mg, about 39 mg, about 40 mg, about 41 mg, about 42 mg, about 43 mg, about 44 mg, about 45 mg, about 46 mg, about 47 mg, about 48 mg, about 49 mg, or about 50 mg. In one embodiment, the compound of formula (I) is administered to the subject at a dose of about 24 mg. In one embodiment, the compound of formula (I) is administered to the subject at a dose of about 16 mg. In one embodiment, the compound of formula (I) is administered to the subject at a dose of about 8 mg. In one embodiment, the compound of formula (I) is administered to the subject at a dose of about 4 mg.

[0043] The anti-PD-1 antibody molecule may be administered at a dose of about 10 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 190 mg, about 200 mg, about 210 mg, about 220 mg, about 230 mg, about 240 mg, about 250 mg, about 260 mg, about 270 mg, about 280 mg, about 290 mg, about 300 mg, about 310 mg, about 320 mg, about 330 mg, about 340 mg, about 350 mg, about 360 mg, about 370 mg, about 380 mg, about 390 mg, or about 400 mg. In one embodiment, the anti-PD-1 antibody molecule is administered to said subject at a dose of about 200 mg.

[0044] Unless otherwise stated or clear from the context, the term "about" as used herein is understood to be within the normal tolerance in the art, for example, within two standard deviations of the mean. About can be understood as within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% of the stated value. Unless otherwise clear from the context, all numerical values ​​provided herein can be modified by the term about.

[0045] The appropriate dosage and administration regimen can be determined by the attending physician and can depend on the severity of the condition as well as the general age, health and weight of the patient being treated.For example, the administration schedule (such as the administration schedule of the compound of formula (I)) can be changed during the course of treatment, such as increasing or decreasing the dosage or frequency during the course of treatment.The administration schedule can also include a break in administration.

[0046] In one embodiment, the method further comprises administering a bone protective agent, which may be denosumab or zoledronic acid or both. The antibody molecule may be administered by various methods known in the art, but for many therapeutic applications, the preferred route / mode of administration is intravenous injection or infusion. Therapeutic compositions for intravenous injection or infusion should usually be sterile and stable under the conditions of manufacture and storage. The composition may be formulated as a solution, microemulsion, dispersion, liposome, or other ordered structure suitable for high antibody concentration. Sterile injectable solutions are prepared by combining the active compound (i.e., antibody or antibody portion) in the required amount in a suitable solvent with one or a combination of ingredients, as appropriate. It can be prepared by sterilization by filtration. Generally, dispersions are prepared by blending the active compound in a sterile solvent containing a basic dispersion medium and other required ingredients. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred method of preparation is vacuum drying and freeze-drying, which obtains a powder of the active ingredient and any additional desired ingredients from a previously sterile-filtered solution thereof. The proper fluidity of the solution can be maintained, for example, by the use of a coating agent such as lecithin, by maintaining the required particle size in the case of dispersions, and by the use of surfactants. Prolonged absorption of injectable compositions can be achieved by including in the composition an agent that delays absorption, for example, monostearate salts and gelatin.

[0047] It will be understood that the route and / or mode of administration will vary according to the desired result.For example, the active compound can be prepared with a carrier that will prevent the rapid release of the compound, such as a controlled release formulation, including implants, transdermal patches, and microencapsulated delivery systems.Biodegradable biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid can be used.Many methods for preparing such formulations are patented or generally known to those skilled in the art (e.g., Sustained and Controlled Release Drug Delivery Systems, edited by JR Robinson, Marcel Dekker, Inc., New York, 1978).

[0048] Disclosed herein are pharmaceutical combinations as defined herein for use as medicaments. Disclosed herein is a pharmaceutical combination as defined herein for use in the treatment of a proliferative disease or disorder associated with the Wnt pathway in a subject.

[0049] Disclosed herein is the use of a pharmaceutical combination as defined herein in the manufacture of a medicament for the treatment of a proliferative disease or disorder associated with the Wnt pathway in a subject.

[0050] Disclosed herein is the use of a compound of formula (I), or a pharma- ceutically acceptable salt or solvate thereof, in the manufacture of a medicament for the treatment of a proliferative disease or disorder associated with the Wnt pathway in a subject, wherein the medicament is administered to a subject in need thereof together with an anti-PD-1 antibody molecule, or a pharma- ceutically acceptable salt or solvate thereof.

[0051] Disclosed herein is the use of an anti-PD-1 antibody molecule, or a pharma- ceutically acceptable salt or solvate thereof, in the manufacture of a medicament for the treatment of a proliferative disease or disorder associated with the Wnt pathway in a subject, wherein the medicament is administered to a subject in need thereof together with a compound of Formula (I), or a pharma- ceutically acceptable salt or solvate thereof.

[0052] Provided herein is a pharmaceutical composition comprising the pharmaceutical combination disclosed herein.In one embodiment, a pharmaceutical composition is provided comprising a compound of formula (I) and an anti-PD-1 antibody.In one embodiment, a pharmaceutical composition is provided comprising a compound of formula (I) and an anti-PD-1 antibody.

[0053] The compositions may contain any suitable carrier, diluent, or excipient, including all conventional solvents, dispersion media, fillers, solid carriers, coating agents, antifungal and antibacterial agents, skin penetration agents, surfactants, isotonic and absorption agents, etc. It will be appreciated that the compositions of the present invention may also contain other auxiliary physiologically active agents.

[0054] The carrier must be pharma- ceutically "acceptable" in the sense of being compatible with the other ingredients of the composition and not harmful to the subject. The composition includes carriers suitable for oral, rectal, nasal, topical (including buccal and sublingual), vaginal, or parenteral (including subcutaneous, intramuscular, intravenous, and intradermal) administration. The composition may conveniently be presented in unit dosage form and may be prepared by any method known in the art of pharmacy. Such methods include the step of mixing the active ingredient with the carrier which constitutes one or more accessory ingredients. In general, the compositions are prepared by uniformly and intimately admixing the active ingredient with liquid carriers or finely divided solid carriers, or both, and then, if necessary, shaping the product.

[0055] Compositions of the present invention suitable for oral administration may be presented as discrete units such as capsules, sachets, or tablets, each containing a predetermined amount of the active ingredient, as a powder or granules, as a solution or suspension in an aqueous or non-aqueous liquid, or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion. The active ingredient may also be presented as a bolus, electuary, or paste.

[0056] Tablets may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the active ingredient in a free-flowing form such as a powder or granules, optionally mixed with a binder (e.g., an inert diluent, a preservative, a disintegrant (e.g., sodium starch glycolate, cross-linked polyvinylpyrrolidone, cross-linked sodium carboxymethylcellulose), a surfactant, or a dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of powdered compound moistened with an inert liquid diluent. The tablets may be optionally coated or scored, and may be formulated to provide sustained or controlled release of the active ingredient therein, for example, using hydroxypropylmethylcellulose in various proportions that provide the desired release profile. Tablets may be optionally enteric coated to provide release in parts of the digestive tract other than the stomach.

[0057] Compositions suitable for topical administration in the mouth include lozenges comprising the active ingredient in a flavored base, usually sucrose and acacia or tragacanth; pastilles comprising the active ingredient in an inert base such as gelatin and glycerin or sucrose and acacia; and mouthwashes comprising the active ingredient in a suitable liquid carrier.

[0058] Compositions suitable for topical administration to the skin may contain the compound dissolved or suspended in any suitable carrier or base and may be in the form of a lotion, gel, cream, paste, ointment, etc. Suitable carriers include mineral oil, propylene glycol, polyoxyethylene, polyoxypropylene, emulsifying wax, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol, and water. Transdermal patches may also be used to administer the compounds of the invention.

[0059] Compositions for rectal administration may be presented as a suppository with a suitable base including, for example, cocoa butter, glycerin, gelatin, or polyethylene glycol. Compositions suitable for vaginal administration may be presented as pessaries, tampons, creams, gels, pastes, foams, or spray formulations containing, in addition to the active ingredient, such carriers as are known in the art to be appropriate.

[0060] Compositions suitable for parenteral administration include aqueous and non-aqueous isotonic sterile injection solutions which may contain antioxidants, buffers, bactericides, and solutes which render the composition isotonic with the blood of the intended recipient, and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents. The compositions may be presented in unit-dose or multi-dose sealed containers, for example, ampoules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example, water for injections, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules, and tablets as described above.

[0061] Preferred unit dosage compositions are those containing a daily dose or unit, daily sub-dose, as herein above recited, or an appropriate fraction thereof, of the active ingredient.

[0062] In addition to the active ingredients specifically mentioned above, it should be understood that the compositions of the present invention may contain other agents conventional in the art considering the type of composition in question, for example, those suitable for oral administration may contain additional agents such as binders, sweeteners, thickeners, flavorings, disintegrants, coatings, preservatives, lubricants, and / or time delay agents. Suitable sweeteners include sucrose, lactose, glucose, aspartame, or saccharin. Suitable disintegrants include corn starch, methylcellulose, polyvinylpyrrolidone, xanthan gum, bentonite, alginic acid, or agar. Suitable flavorings include peppermint oil, oil of wintergreen, cherry, orange, or raspberry flavoring. Suitable coatings include polymers or copolymers of acrylic acid and / or methacrylic acid and / or their esters, waxes, fatty alcohols, zein, shellac, or gluten. Suitable preservatives include sodium benzoate, vitamin E, alpha-tocopherol, ascorbic acid, methylparaben, propylparaben, or sodium bisulfite. Suitable lubricants include magnesium stearate, stearic acid, sodium oleate, sodium chloride, or talc. Suitable time delay agents include glyceryl monostearate or glyceryl distearate.

[0063] Throughout this specification and the following description, unless the context otherwise requires, the word "comprise" and variations such as "comprises" and "comprising" will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.

[0064] As used in the subject specification, the singular forms "a," "an," and "the" include plural aspects unless the context clearly dictates otherwise. Thus, for example, reference to a "formulation" includes two or more formulations in addition to a single formulation, reference to an "agent" includes two or more agents in addition to a single agent, reference to "the disclosure" includes single and multiple aspects taught by that disclosure, etc. Aspects taught and enabled herein are encompassed by the term "invention." All aspects are enabled within the scope of the invention.

[0065] Those skilled in the art will understand that the invention described herein is susceptible to variations and modifications other than those specifically described. It is to be understood that the invention includes all such variations and modifications that fall within its spirit and scope. The invention also includes all steps, features, compositions, and compounds mentioned or suggested herein, individually or collectively, and any and all combinations of any two or more of said steps or features.

[0066] Without being limited thereto, the present invention includes the following aspects. [Aspect 1] a) Formula (I) having the following structure:

number

[0067] Example 1 Preparation of Compounds of Formula (I) Synthesis of amines Suzuki Method A: A stirred solution of aryl halide (1 equiv.), boronic acid (1.5 equiv.), and sodium carbonate (2 equiv.) in toluene (0.08 M) and water (0.32 M) was degassed with argon for 15 min. Tetrakis(triphenylphosphine)palladium(0) (0.05 equiv.) was added to the reaction mixture and the reaction mixture was heated to reflux for 16 h. After completion of the reaction of the starting material, the reaction mixture was concentrated, water was added to the reaction mixture and extracted with ethyl acetate. The combined organic layers were washed with brine, dried with Na2SO4, and concentrated under reduced pressure. The crude compound was purified by column chromatography to give the purified product.

[0068] Synthesis of 6-phenylpyridazin-3-amine.

[0069] [ka]

[0070] 1H NMR (400 MHz, DMSO-d6) δ (ppm): 7.96-7.94 (d, J=8 Hz, 2H), 7.82-7.80 (d, J=9.2 Hz, 1H), 7.48-7.35 (m, 3H), 6.86-6.84 (m, 1H), 6.64(br s, 2H).LC-MS: Pure m / z 172.0 (M+H), purity 82%.

[0071] Amide coupling method A: To a stirred solution of commercially available 2-(1,3-dimethyl-2,6-dioxo-2,3-dihydro-1H-purin-7(6H)-yl)acetic acid, 1 (1 eq.) in dichloromethane (0.01 M), HATU (1.3 eq.), triethylamine (1.5 eq.), and the respective amine (1 eq.) were added. The reaction mixture was stirred at room temperature. Immediately after completion of the reaction, water was added to the reaction mixture and the mixture was extracted with dichloromethane. The organic layer was washed with brine solution, dried over anhydrous Na2SO4, and concentrated under reduced pressure to give the crude product. The crude product was further purified by column chromatography.

[0072] 2-(1,3-dimethyl-2,6-dioxo-1,2,3,6-tetrahydro-7H-purin-7-yl)-N-(6-phenylpyridazin-3-yl)acetamide (also known as 2-(1,3-dimethyl-2,6-dioxo-2,3-dihydro-1H-purin-7(6H)-yl)-N-(6-phenylpyridazin-3-yl)acetamide).

[0073] [ka]

[0074] 1 H NMR (400 MHz, DMSO-d6) δ (ppm): 11.77 (s, 1H), 8.32-8.25 (m, 2H), 8.12-8.10 (m, 3H), 7.57-7.49 (m, 3H), 5.37 (s, 2H), 3.46 (s, 3H), 3.19 (s, 3H).LC-MS : m / z 390.1 (M+H) with 99% purity.

[0075] Example 2 The single-agent activity of Compound A increased cell surface PD-1 expression and enhanced upregulation of genes that make tumors "hotter."

[0076] FFPE tumor tissue from patient 202-101 with a tumor of unknown primary site (probable cholangiocarcinoma) was prepared from paired pre- and post-C1D17 biopsies. Sections were evaluated via IHC / OPAL™ and analyzed on a Vectra3 (Perkin Elmer). Cells in each high-power field at 400x magnification were counted on the Vectra3 and the mean ± SD from a minimum of n=3 different fields was graphed. Numbers obtained from single stainings were used to determine FOXP3 + CD8 on cells + Cells and FOXP3 + CD4 on cells + The percentage of cells was plotted (Figure 1).

[0077] As a single agent, Compound A altered the expression levels of immune environment genes, making tumors hotter. Nanostring analysis of FFPE and frozen tumor tissues from patient 202-101 (T4-T3) with tumor of unknown primary site (probable cholangiocarcinoma) and MSS CRC102-102 (T2-T1) patient (with RSPO gene fusion) using a pan-cancer immunopanel. (Figure 2A) Grouped analysis showed similar pre- and post-treatment changes in both specimens. (Figure 2B) The most up- and down-regulated genes common to both cancer types are shown. (Figure 2C and Figure 2D) The most up- and down-regulated genes in a CRC patient with RSPO fusion and a patient with unknown primary tumor (probable cholangiocarcinoma), respectively, are shown. Red gene symbols indicate genes that were up- or downregulated in both patients.

[0078] Example 3 Dose Escalation Study A 3+3 dose escalation study of Compound A with standard dose pembrolizumab will be conducted first. In this study, various doses of Compound A (e.g., 8 mg, 16 mg, or 24 mg) will be administered simultaneously with pembrolizumab (e.g., 200 mg) to determine the recommended dose (RD) of Compound A for dose expansion.

[0079] Example 4 Dose Expansion Study Main purpose To evaluate the safety of Compound A administered orally every other day at a recommended dose (RD) of 16 mg (which may be adjusted to 8 mg or 24 mg depending on the results of a dose escalation study) as a single agent until disease progression and then in combination with pembrolizumab (a PD-1 inhibitor) in two groups of patients with advanced or metastatic or unresectable solid malignancies that are refractory, intolerant, or unsuitable for available treatments according to the treating physician, as measured by adverse event (AE) monitoring, imaging assessments, 12-lead electrocardiogram readings, Eastern Cooperative Oncology Group performance status, clinical laboratory results, and vital sign measurements.

[0080] To evaluate the tolerability of Compound A administered orally every other day at RD 16 mg as a single agent and in combination with pembrolizumab (a PD-1 inhibitor). Tolerability assessments were performed in two groups (Group 1 and Group 2 [subgroups 2a, 2b, and 2c]) of patients with advanced or metastatic or unresectable solid malignancies who are refractory to, intolerant of, or unsuitable for available treatments according to the treating physician. This will be accompanied by monitoring for fatigue and gastrointestinal symptoms, and immune-related side effects, if applicable. Patient population Compound A (compound of formula (I)) at the recommended dose as a single agent until progression and then in combination with pembrolizumab (a PD-1 inhibitor) with or without bone-protective treatment (denosumab and then zoledronic acid if denosumab fails to respond). Patients with advanced or metastatic or unresectable solid malignancies that are refractory, intolerant or unsuitable to available treatments according to the treating physician will be divided into two groups.

[0081] Group 1 will consist of up to 20 patients with MSS colorectal cancer (CRC) who will be preselected for the presence of one of four different recurrent gene fusions involving R-spondin (RSPO)2 or RSPO3 (fusions A-D) using the D3 RSPO gene fusion test.

[0082] Group 2 will consist of up to 45 patients divided into three subgroups. - Subgroup 2a (N=15): Patients with MSS-CRC who will be preselected for the absence of all four different recurrent gene fusions involving RSPO2 or RSPO3 (fusions A-D) using the D3 RSPO gene fusion test; - Subgroup 2b (N=15): Patients with MSS ovarian cancer who will be recruited without preselection (because of additional mutations), and - Subgroup 2c (N=15): Patients with MSS endometrial cancer who will be recruited without preselection (due to additional mutations). Study design Preliminary efficacy and safety data will be collected during treatment with the recommended dose of 16 mg of compound A administered orally once every other day as a single agent with or without bone-protective treatment (denosumab and then zoledronic acid if denosumab fails to respond) and in combination with pembrolizumab (a PD-1 inhibitor) after progression in an open-label, non-randomized study. Completion of two cycles (6 weeks) of compound A monotherapy is recommended before adding pembrolizumab treatment. However, completion of a minimum of one cycle (3 weeks) of compound A monotherapy before adding pembrolizumab is acceptable. Patients who do not complete the minimum period of one cycle (21 days) of compound A monotherapy for reasons other than safety may be replaced. The actual dosing schedule and frequency of bone-protective treatment will be based on observed serum levels of a bone resorption biomarker (β-CTX). period Single Agent Treatment Period (Dose Expansion) - Compound A administered until disease progression per RECIST version 1.1, initiation of new cancer therapy, or completion of 12 cycles (1 cycle: 21 days) of treatment (if patient did not enter the pembrolizumab combination treatment period).

[0083] Combination Treatment Period - Compound A and pembrolizumab administered until disease progression according to iRECIST, a new cancer therapy is initiated, or 12 cycles (1 cycle: 21 days) of treatment are completed. In either the single agent or combination treatment period, patients who have completed 12 cycles (1 cycle: 21 days) of treatment may continue treatment until disease progression, unacceptable toxicity, patient withdrawal, or loss of clinical benefit, as agreed with the sponsor. Investigational drug, dose, and route of administration (dose expansion) All patients will receive Compound A at the recommended dose of 16 mg as a single agent until disease progression, and then in combination with pembrolizumab until disease progression or loss of clinical benefit. Pembrolizumab will be administered at a dose of 200 mg (administered intravenously over 30 minutes). Compound A will be administered at 8 mg every other day for 21 days at 1 mg / kg / day. If not tolerated, Compound A may be reduced to 8 mg every other day at any time during the study. Bone protection treatment, dosage, and route of administration: Denosumab will be administered SC every 28 days at a starting dose of 120 mg if serum levels of β-CTX are above 1000 pg / mL. If after 1 week of denosumab (SC) treatment, β-CTX levels do not fall below 1000 pg / mL, denosumab will be discontinued and zoledronic acid will be initiated at a dose of 4 mg intravenously every 3-4 weeks. If after administration of denosumab and zoledronic acid (sequentially), β-CTX levels do not fall below 1000 pg / mL within 3 weeks (3 consecutive assessments), the patient will be discontinued from the study. During denosumab or zoledronic acid treatment, patients will additionally receive calcium ≥ 1000 mg orally daily and vitamin D (at least 400 IU) orally daily, based on the investigator's clinical judgment.

Claims

1. (a) Formula (I) having the following structure: 【Chemistry 1】 or a pharma- ceutically acceptable salt or solvate thereof, (b) an anti-PD-1 antibody molecule, or a pharma- ceutically acceptable salt or solvate thereof; A pharmaceutical combination comprising:

2. The combination of claim 1, wherein the anti-PD-1 antibody molecule is pembrolizumab or a fragment thereof.

3. 2. The pharmaceutical combination of claim 1 for use in the treatment of a proliferative disease or disorder associated with the Wnt pathway in a subject.

4. 4. The combination of claim 3, wherein the proliferative disease or condition is cancer, fibrosis, stem cell and diabetic retinopathy, rheumatoid arthritis, psoriasis, or myocardial infarction.

5. 5. The combination of claim 3 or claim 4, wherein the proliferative disease or condition is lung cancer, prostate cancer, melanoma, osteosarcoma, colorectal cancer, or endometrial cancer.

6. 5. The combination of claim 3 or claim 4, wherein the proliferative disease or condition is selected from the group consisting of cervical cancer, colon cancer, breast cancer, bladder cancer, head and neck cancer, gastric cancer, lung cancer, ovarian cancer, prostate cancer, thyroid cancer, non-small cell lung cancer, chronic lymphocytic leukemia, mesothelioma, melanoma, pancreatic adenocarcinoma, basal cell carcinoma, osteosarcoma, hepatocellular carcinoma, Wilms' tumor, medulloblastoma, pulmonary fibrosis, liver fibrosis, skin fibrosis, renal fibrosis, stem cell and diabetic retinopathy, rheumatoid arthritis, psoriasis, and myocardial infarction.

7. 7. The combination according to any one of claims 3 to 6, wherein the compound of formula (I) is administered to the subject once daily.

8. The combination of any one of claims 3 to 7, wherein the anti-PD-1 antibody molecule is administered to the subject once every 21 days.

9. 9. The combination according to any one of claims 3 to 8, wherein the compound of formula (I) is administered orally to the subject.

10. The combination of any one of claims 3 to 9, wherein the anti-PD-1 antibody molecule is administered to the subject by intravenous infusion.

11. 11. The combination according to any one of claims 3 to 10, wherein the compound of formula (I) is administered to the subject in a dose of about 8 mg, about 16 mg, or about 24 mg.

12. The combination of any one of claims 3 to 11, wherein the anti-PD-1 antibody molecule is administered to the subject at a dose of about 200 mg.

13. 13. Use of the pharmaceutical combination of claim 1 in the manufacture of a medicament for the treatment of a proliferative disease or disorder associated with the Wnt pathway in a subject.

14. 14. The use according to claim 13, wherein the proliferative disease or condition is cancer, fibrosis, stem cell and diabetic retinopathy, rheumatoid arthritis, psoriasis, or myocardial infarction.

15. 15. The use according to claim 13 or claim 14, wherein the proliferative disease or condition is lung cancer, prostate cancer, melanoma, osteosarcoma, colorectal cancer, or endometrial cancer.

16. 15. The use of claim 13 or claim 14, wherein the proliferative disease or condition is selected from the group consisting of cervical cancer, colon cancer, breast cancer, bladder cancer, head and neck cancer, gastric cancer, lung cancer, ovarian cancer, prostate cancer, thyroid cancer, non-small cell lung cancer, chronic lymphocytic leukemia, mesothelioma, melanoma, pancreatic adenocarcinoma, basal cell carcinoma, osteosarcoma, hepatocellular carcinoma, Wilms' tumor, medulloblastoma, pulmonary fibrosis, liver fibrosis, skin fibrosis, renal fibrosis, stem cell and diabetic retinopathy, rheumatoid arthritis, psoriasis, and myocardial infarction.

17. 17. The use according to any one of claims 13 to 16, wherein the compound of formula (I) is administered to the subject once daily.

18. The use of any one of claims 13 to 17, wherein the anti-PD-1 antibody molecule is administered to the subject once every 21 days.

19. 19. The use according to any one of claims 13 to 18, wherein the compound of formula (I) is administered orally to the subject.

20. The use according to any one of claims 13 to 19, wherein the anti-PD-1 antibody molecule is administered to the subject by intravenous infusion.

21. 21. The use according to any one of claims 13 to 20, wherein the compound of formula (I) is administered to the subject at a dose of about 8 mg, about 16 mg, or about 24 mg.

22. 22. The use of any one of claims 13 to 21, wherein the anti-PD-1 antibody molecule is administered to the subject at a dose of about 200 mg.

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