HK40000167A - Combination of a chromene compound and a second active agent - Google Patents

Description

Combination of a chromene compound and a second active agent

Cross Reference to Related Applications

This application is based on the benefit of 35U.S. c. § 119(e) claiming U.S. application No. 62/276,713, filed 2016 at 8/1 and U.S. application No. 62/277,225, filed 2016 at 11/1. The contents of application nos. 62/276,713 and 62/277,225 are incorporated by reference herein in their entirety.

FIELD

The present disclosure generally relates to a combination of a pharmaceutically acceptable chromene compound and a second compound, pharmaceutical compositions comprising said combination, and methods of treating an individual by administering said combination. More particularly, the disclosure relates to combinations comprising a class of deuterated and non-deuterated chromene compounds and a second active compound, and methods of preventing and treating various cancers.

Background

This section provides background information related to the present disclosure, which is not necessarily prior art.

Nonsteroidal anti-inflammatory drugs (NSAIDs) have been demonstrated to treat and prevent various cancers. Non-selective NSAIDs inhibit both COX-1 and COX-2. COX-2 contributes to carcinogenesis by increasing prostanoid production, inhibiting apoptosis, promoting angiogenesis, and regulating inflammation and immune function. COX-2 inhibitors may be effective in treating various cancers.

Some selective COX-2 inhibitors contain chromene structural features. Chromene-based COX-2 inhibitors have selectivity and analgesic potency similar to those of the diaryl heterocyclic coxib compounds. However, unlike the diaryl heterocyclic coxib-based drugs, chromene-based COX-2 inhibitors do not damage the kidney, thereby reducing the likelihood of hypertension.

Urine PGE-M is the major urinary metabolite of PGE2 and can be used as an index for systemic PGE2 production. Both non-selective NSAIDs and COX-2 selective inhibitors inhibit PGE-M levels. Given that the anti-tumor effects of NSAIDs (e.g., COX-2 inhibitors) are dependent on the reduction of PGE2 production by targeting COX-2, urinary PGE-M serves as a valuable intermediate marker for the pharmacological activity of NSAIDs in the prevention and treatment of cancer. PGE-M is a potent biomarker for predicting the effect of COX-2 inhibitors in patients with COX-2 overexpression dependent cancers (Wang et al, Cancer Prev.Res., 2013).

WO 03/015608 describes methods of treating or preventing cancer using protein kinase inhibitors that can bind to COX-2 inhibitors.

US 2004/0127470 describes methods of treating neoplastic disorders using a combination of a COX-2 inhibitor and an EGFR antagonist.

WO2013/189121 reports novel deuterated benzopyran compounds having anti-inflammatory and anti-tumor effects.

Zhang et al (ACS med. chem.lett.2015) describes a method of treating lung cancer in vitro using erlotinib covalent linkers linked to different NSAIDs.

SUMMARY

This section provides a general summary of the disclosure, and is not a comprehensive disclosure of its full scope or all of its features.

Provided herein are combinations of a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, wherein formula (I) is

Wherein M is selected from H and alkyl;

z is selected from CF₃、CF₂H and C₂F₅；

Each R is¹、R²、R³And R⁴Independently selected from the group consisting of H, alkyl, aralkyl, deuterated alkyl, deuterated aralkyl, deuterated alkoxy, deuterated cycloalkyl, deuteron, deuteroaryloxy, deuteroheteroaryloxy, deuteroarylalkoxy, deuteroheteroarylalkoxy, deuterohaloalkoxy, deuteroamino, deuterosulfonamido, sulfonamido, cycloalkyl, cycloalkenyl, halogen, haloalkyl, alkoxy, haloalkoxy, alkylthio, haloalkylthio, pentafluorosulfanyl, hydroxyalkyl, trialkylsilyl, alkynyl, and alkenyl; and

wherein the second compound is selected from the group consisting of a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, an OX-40 agonist, a CD137 agonist, a LAG-3 inhibitor, an IDO inhibitor, a bispecific protein, an EGFR inhibitor, a HER2 inhibitor, and an immunostimulatory therapeutic agent.

In another embodiment, a pharmaceutical composition is provided comprising a therapeutically effective amount of a combination of a compound of formula (II) and a second compound.

In another embodiment, there is provided a method of treating cancer comprising administering to a subject in need thereof a therapeutically effective amount of a compound of formula (I) and a second compound in combination.

Brief Description of Drawings

FIG. 1 shows tumor volume (mm) for compound A01, erlotinib, or a combination thereof in mice inoculated with colorectal cancer cells³) The function of (1).

Figure 2 shows the effect of compound a01, an anti-PD 1 antibody, or a combination thereof on tumor rejection in mice inoculated with colon cancer cells.

FIG. 3 shows a CD8⁺Linear regression in which the growth of T cells is associated with a decrease in tumor volume.

FIG. 4 shows CD8 of Compound A01, an anti-PD 1 antibody, or a combination thereof against mice injected with colon cancer cells⁺Effects on the T cell level.

Detailed description of the invention

A. Definition of

The term "deuterium" as used herein means a single deuterium atom wherein the deuterium radical is attached to a carbon to form a deuterated compound.

As used herein, the terms "alkyl" and "alkylene" refer to branched and straight chain saturated aliphatic hydrocarbon groups having the indicated number of carbon atoms. For example, "C₁-C₅C in alkyl₁-C₅"is meant to include groups having 1, 2,3, 4, or 5 carbon atoms in a straight or branched chain arrangement. For example, "C₁-C₅The alkyl group "specifically includes methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, pentyl and the like.

The term "cycloalkyl" means a monocyclic saturated aliphatic hydrocarbon group having the specified number of carbon atoms. For example, "cycloalkyl" includes cyclopropyl, methyl-cyclopropyl, 2-dimethyl-cyclobutyl, 2-ethyl-cyclopentyl and cyclohexyl.

As used herein, the terms "alkenyl" and "alkenylene" refer to branched and straight chain unsaturated or partially unsaturated hydrocarbon groups having the specified carbon number and at least one carbon-carbon double bond. The term "cycloalkenyl" means a monocyclic unsaturated or partially unsaturated aliphatic hydrocarbon group having the specified number of carbon atoms and at least one carbon-carbon double bond.

As used herein, the terms "alkynyl" and "alkynylene" refer to branched and straight chain unsaturated or partially unsaturated hydrocarbon groups having the specified carbon number and at least one carbon-carbon triple bond.

The term "alkoxy" as used herein denotes a cyclic or acyclic alkyl group of the indicated number of carbon atoms attached through an oxygen bridge. Thus, "alkoxy" encompasses the above definitions of alkyl and cycloalkyl.

The term "aryl" as used herein means any stable monocyclic or bicyclic carbocyclic ring of up to 7 atoms in each ring, wherein at least one ring is aromatic. Examples of such aryl elements include phenyl, naphthyl, tetrahydronaphthyl, indanyl, and biphenyl.

The term "heteroaryl" as used herein means a stable monocyclic or bicyclic ring of up to 7 atoms in each ring, wherein at least one ring is aromatic and contains 1 to 4 heteroatoms selected from O, N and S. Heteroaryl groups within the scope of this definition include, but are not limited to: acridinyl, carbazolyl, cinnolinyl, quinoxalinyl, pyrazolyl, indolyl, benzotriazolyl, furanyl, thienyl, benzothienyl, benzofuranyl, quinolinyl, isoquinolinyl, oxazolyl, isoxazolyl, indolyl, pyrazinyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, and tetrahydroquinoline.

The term "halo" or "halogen" as used herein includes chloro, fluoro, bromo and iodo.

The present disclosure includes compounds of formula (I) in free form, as well as pharmaceutically acceptable salts and stereoisomers thereof. Some specific compounds exemplified herein are protonated salts of amine compounds. The term "free form" refers to the non-salt form of the chromene compound. Encompassed pharmaceutically acceptable salts include not only the salts exemplified for the specific compounds described herein, but also all typical pharmaceutically acceptable salts of the compounds of formula (I) in free form. The free form of the particular salt compound described can be isolated using techniques known in the art.

Pharmaceutically acceptable salts of the present disclosure can be synthesized from compounds of the present disclosure containing a basic or acidic moiety by conventional chemical methods.

When a compound of the present disclosure is acidic, suitable "pharmaceutically acceptable salts" refer to salts prepared from pharmaceutically acceptable non-toxic bases including inorganic and organic bases.

It should be noted that the compounds of the present disclosure may be internal salts or zwitterions, as under physiological conditions, the deprotonated acidic moiety (e.g., carboxyl group) in the compound may be anionic, and this electronic charge may be balanced internally with the cationic charge of the protonated or alkylated basic moiety (e.g., quaternary nitrogen atom).

The term "combination" as used herein means the use of both the chromene compound of formula (I) or formula (II) and the second compound described herein. The combination includes co-presentation of the two compounds, e.g., a kit or co-packaged product, and use by the patient of the two compounds to obtain or prescribe separately. The chromene compound of formula (I) or formula (II) in the combination may be administered to the patient before, concomitantly with, after or in alternation with the second compound.

Immune checkpoint proteins are components of the immune system that can be used to stimulate or inhibit immune signals (e.g., T cell activation signals) in general. Non-limiting examples of stimulatory immune checkpoint proteins include CD27, CD40, OX40, GITR, CD137, CD28, HVEM, and ICOS. Non-limiting examples of inhibitory immune checkpoint proteins include adenosine A_2AReceptors, B7-H3, B7-H4, BTLA, CTLA-4, IDO, KIR, LAG-3, PD-1, PD-L1, TIM-3 and VISTA (C10 or f 54).

An immune checkpoint modulating compound is any compound that modulates the activity of an immune checkpoint protein, such as an antibody, a small molecule, a biologic, or a polysaccharide. Modulation of an immune checkpoint protein may include the activity of the compound as an agonist, antagonist, allosteric effector, or any effect from binding to an immune checkpoint protein or protein ligand that alters the general biological function of the immune checkpoint protein. Many cancers express high levels of inhibitory immune checkpoint proteins in order to circumvent the detection of T cells and other immune system components. Blocking an inhibitory immune checkpoint protein (e.g., PD-1) using an antagonist antibody enhances the immune response that detects and destroys cancer cells. Conversely, activation of a stimulatory immune checkpoint protein (e.g., OX-40) using an agonist antibody enhances the immune response that recognizes and destroys cancer cells.

B. Chromene compounds

The present disclosure relates to chromene compounds having the structure represented by formula (I) or formula (II). The compound and the pharmaceutically acceptable salt thereof disclosed by the application can be applied to preparation of anti-inflammatory and analgesic drugs and drugs for treating or preventing tumors.

In one embodiment, the present disclosure provides a combination comprising a chromene compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof, and a second compound:

wherein M is selected from H and alkyl; z is selected from-CF₃、-CF₂H and-C₂F₅(ii) a Each R is¹、R²、R³And R⁴Independently selected from the group consisting of H, alkyl, aralkyl, deuterated alkyl, deuterated aralkyl, deuterated alkoxy, deuterated cycloalkyl, deuteron, deuteroaryloxy, deuteroheteroaryloxy, deuteroarylalkoxy, deuteroheteroarylalkoxy, deuterohaloalkoxy, deuteroamino, deuterosulfonamido, sulfonamido, cycloalkyl, cycloalkenyl, halo, haloalkyl, alkoxy, haloalkoxy, alkylthio, haloalkylthio, pentafluorosulfanyl, hydroxyalkyl, trialkylsilyl, alkynyl and alkenyl.

When any variable (e.g., R)¹Z, etc.) in any componentWhen occurring more than one time, its definition at each occurrence is independent of every other occurrence. Also, combinations of substituents and variables are only allowed when these combinations result in stable compounds. The line drawn into the ring system from a substituent indicates that the bond shown may be attached to any substitutable ring atom. If the ring system is polycyclic, it means that the bond is only attached to any suitable carbon atom on the adjacent ring. It is to be understood that substituents and substitution patterns on the compounds of the present invention can be selected by one of ordinary skill in the art to provide compounds that are chemically stable and can be readily synthesized from readily available starting materials by techniques known in the art as well as those methods described below.

The chromene compound of formula (I) can be prepared by using the following reaction, in addition to methods which have been disclosed in the literature or well-proven in experimental procedures. Thus, the following synthetic schemes are merely illustrative and are not intended to limit the compounds or any particular substituents. The number of substituents in a scheme need not correspond to the number defined in the claims. Furthermore, for clarity, compounds of formula (I) or formula (II) showing single substitution may allow compounds with multiple substituents.

In one embodiment, when Z ═ CF₃When the chromene of formula (I) can be prepared by reaction of salicylaldehyde (prepared from the corresponding phenol; see WO 2013/189121; CN 102757417; CN 103044477; and CN 103012350; each of which is incorporated herein by reference) with ethyl 4,4, 4-trifluorocrotonate according to procedures described in the literature (i.e., U.S. patent No. 6,034,256), or when Z ═ CF₂CF₃When using 4,4,5,5, 5-pentafluoro-but-2-enoic acid ethyl ester (CAS # [37759-78-7 ])]) To prepare the compound. Alternatively, where Z ═ CF is prepared by the reaction of salicylaldehyde with 4,4, 4-trifluorocrotonaldehyde and a chiral catalyst followed by oxidation according to the procedure described in ACS med₃The chiral chromenic acid of (1). Preparation of the compound wherein Z-CF is prepared by a similar method using 4,4,5,5, 5-pentafluoropent-2-enal₂CF₃The 4,4,5,5, 5-pentafluoropent-2-enalPrepared from 4,4,5,5, 5-pentafluoropent-2-en-1-ol using the same procedure described below for the preparation of 4,4, 4-trifluorocrotonaldehyde (INT-03).

In various embodiments, the compounds of formula (I) have at least one deuterated substituent, e.g., deuterated alkyl, deuterated cycloalkyl, and deuteron. In one embodiment, R¹、R²、R³And R⁴At least one of which is deuterated alkyl, deuterated cycloalkyl or deuteron. In some embodiments, the chromene compound in the combination has the structure depicted in formula (II), or a pharmaceutically acceptable salt or stereoisomer thereof, or a prodrug molecule thereof:

wherein X is selected from O, S and NR^a；

R^aSelected from H, C₁-C₃Alkyl radical, C₃-C₆Cycloalkyl, C substituted by one or two halogens₁-C₃Alkyl, and aryl;

r is selected from carboxyl, amido, alkyl sulfonyl, C₁-C₃C substituted by cyclocarbonyl or aryl₁-C₃Cyclic carbonyl and C₁-C₃Alkoxycarbonyl groups, and alkoxycarbonyl groups;

R¹selected from haloalkyl, alkyl, aralkyl, phenyl, and cycloalkyl;

R²one or more selected from the following groups: hydrogen, deuterium, halogen, alkyl, deuterated alkyl, aralkyl, deuterated aralkyl, alkoxy, deuterated alkoxy, aryloxy, deuterated aryloxy, heteroaryloxy, deuterated heteroaryloxy, arylalkoxy, deuterated arylalkoxy, heteroarylalkoxy, deuterated heteroarylalkoxy, halogenated alkoxy, deuterated halogenated alkoxy, amino, deuterated amino, sulfonamido, pentafluorosulfanylAnd deuterated sulfonamido; and

n is an integer selected from 1, 2 and 3.

R³Is deuterated alkyl.

Specific embodiments of the chromene compounds of formula (I) include:

table 1: deuterated compounds

Table 2: non-deuterated compounds

In another embodiment, the chromene compound of formula (II):

wherein X is O, R^aSelected from H, C₁-C₃Alkyl radical, C₃-C₆Cycloalkyl, C substituted by one or two halogens₁-C₃Alkyl, and aryl; n is an integer selected from 1, 2 and 3; r is selected from carboxyl and alkoxycarbonyl; r¹Selected from haloalkyl, alkyl, aralkyl, and cycloalkyl; each R is²Independently selected from deuterium, halogen, alkyl, deuterated alkyl, aralkyl, deuterated aralkyl, haloalkyl, deuterated haloAlkyl, alkoxy, deuterated alkoxy, aryloxy, deuterated aryloxy, heteroaryloxy, deuterated heteroaryloxy, arylalkoxy, deuterated arylalkoxy, heteroarylalkoxy, deuterated heteroarylalkoxy, haloalkoxy, deuterated haloalkoxy, amino, deuterated amino, sulfonamido, and deuterated sulfonamido; and R is³Is deuterated alkyl. In another embodiment, position 7 is unsubstituted; r is carboxyl or C₁-C₃An alkoxycarbonyl group; r¹Is a haloalkyl group; and combinations thereof. In another embodiment, n is 1 or 2; r is carboxyl or C₁-C₃An alkoxycarbonyl group; r¹Is haloalkyl, cycloalkyl or phenyl; r²Is deuterium, halogen, alkyl, deuterated alkyl, haloalkyl, deuterated haloalkyl, alkoxy, deuterated alkoxy, alkylamino, deuterated alkylamino, alkylated sulfonamido, and alkylated deuterated sulfonamido; or a combination thereof, and R²At least one of the substitutions is in position 6.

C. Second compound

The chromene compound of formula (I) is combined with a second compound. The second compound is a small molecule, drug, peptide, antibody, or pharmaceutical agent. In one embodiment, the second compound can be a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, an OX-40 agonist, a CD137 agonist, a LAG-3 inhibitor, an IDO inhibitor, a bispecific protein, an EGFR inhibitor, a HER2 inhibitor, or an immunostimulatory therapeutic.

In one embodiment, the second compound is a PD-1 inhibitor. PD-1 inhibitors are immune checkpoint modulators that act on the immune checkpoint protein programmed cell death protein 1, also known as cluster of differentiation 279(CD 279). PD-1 is present on immune cells and is commonly used as an "off-switch" to prevent T cell activation. This inhibitory function is activated when PD-1 binds to PD-L1, and PD-L1 is present on many tumors. In some embodiments, the PD-1 inhibitor is selected from the group consisting of nivolumab (nivolumab), pidilizumab, parbolizumab (pembrolizumab), AMP-224(CAS #1422184-00-6), AMP-514(MEDI0680, CAS #1642374-69-3), STI-A1110, TSR-043, and AUNP-12(AUR-012, Aurigene-012, Aurigene NP-12).

In one embodiment, the second compound is a PD-L1 inhibitor. PD-L1 inhibitors are immune checkpoint modulators that act on the immune checkpoint protein programmed death ligand 1, also known as cluster of differentiation 274(CD274) or B7 homolog 1 (B7-H1). High expression of PD-L1 has been shown to be associated with increased tumor aggressiveness and lower survival rates, as upregulation of PD-L1 in tumors may allow tumors to evade the immune system. This phenomenon occurs through the binding of PD-L1 to PD-1, and the PD-L1 inhibitor prevents this by binding to PD-L1. In some embodiments, the PD-L1 inhibitor is selected from RG 7446, BMS-936559(MDX 1105, CAS #1422185-22-5), MSB0010718C, STI-a1010, avizumab (avelumab), alezumab (atezolizumab), and covaptumab (durvalumab).

In one embodiment, the second compound is a CTLA-4 inhibitor. CTLA-4 inhibitors are immune checkpoint modulators that act on the immune checkpoint protein, cytotoxic T-lymphocyte-associated protein 4, also known as cluster of differentiation 152(CD 152). CTLA-4 checkpoint proteins are expressed in activated T cells and tregs, and binding of CTLA-4 to CD80 or CD86 inhibits immune function. CTLA-4 can act by either competing against CD28 or by preventing CD28 from binding to CD80 or CD86, which elicit an immunostimulatory effect. CTLA-4 can function by capturing and removing CD80 and CD86 from antigen presenting cells. In some embodiments, the CTLA-4 inhibitor is selected from ipilimumab (ipilimumab) and tremelimumab.

In one embodiment, the second compound is an OX-40 agonist. OX-40 agonists are immune checkpoint modulators that act on the immune checkpoint protein, tumor necrosis factor receptor superfamily member 4(TNFRSF4), also known as cluster of differentiation 134(CD134) and OX-40. OX-40 is a secondary costimulatory immune checkpoint protein, expressed 24 to 72 hours after immune activation. As OX-40L binds to OX-40 receptors on T cells, preventing T cell death, and increasing cytokine production, OX-40 is critical to maintaining the immune response over the first few days until the memory response. In some embodiments, the OX40 agonist is selected from the group consisting of anti-OX 40, TIM3 antibody, and immitune IMP 701.

In one embodiment, the second compound is a CD137 agonist. CD137 agonists are immune checkpoint modulators and include any compound that binds to the CD137 protein, including but not limited to antibodies and small molecules. CD137 is also known as tumor necrosis factor receptor superfamily member 9(TNFRSF9), 4-1BB, and is introduced by lymphocyte activation (ILA), which causes stimulation of the immune system. CD137 may be expressed by activated T cells, but is expressed to a greater extent on CD8 than on CD 4T cells. In addition, CD137 expression was found on dendritic cells, B cells, follicular dendritic cells, natural killer cells, granulocytes, and vascular wall cells at sites of inflammation. In some embodiments, the CD137 agonist is selected from urelumab and utomicumab.

In one embodiment, the second compound is a LAG-3 inhibitor. LAG-3 inhibitors are immune checkpoint modulators and include any compound that binds LAG-3 protein and prevents its inhibitory effect on the immune system, including but not limited to antibodies and small molecules. The primary ligand of LAG-3 is MHC class II, which binds this ligand with higher affinity than CD 4. The protein down-regulates cell proliferation, activation and homeostasis of T cells in a similar manner to CTLA-4 and PD-1, and has been reported to have an important role in Treg suppressive function. LAG-3 also helps to convert CD8⁺T cells are maintained in a tolerogenic state and, in cooperation with PD-1, help to maintain CD8 depletion during chronic viral infection. In some embodiments, the LAG-3 inhibitor is BMS-986016(CAS # 1683572-29-3).

In one embodiment, the second compound is an IDO inhibitor. IDO inhibitors are immune checkpoint modulators and include any compound that binds to indoleamine 2, 3-dioxygenase and prevents it from sending inhibitory signals to the immune system, including but not limited to antibodies and small molecules. IDO can allow tumor cells to evade the immune system by consuming L-Trp in the microenvironment of the cell. There are a wide range of human cancers, such as prostate, colorectal, pancreatic, cervical, gastric, ovarian, head, lung, etc., that overexpress human IDO (hIDO). In some embodiments, the IDO inhibitor is selected from GDC-0919(CAS #1402836-58-1), indoximod, 1-methyl-D-tryptophan (NSC-721782), NLG919(CAS #1402836-58-1) epacadostat, and norharpagne.

In one embodiment, the second compound is a bispecific protein containing at least two domains to bind at least two targets in order to reduce distance or elicit a biological response or both. In another embodiment, the activity of the bispecific protein elicits an enhanced immune response by stimulating the response or preventing the inhibitory effect, or both. Bispecific proteins can bind to epitopes on immune cells, including but not limited to T cells or Natural Killer (NK) cells, as well as tumor cells. The action of the bispecific protein may cause an enhancement of the immune response due to the proximity of immune cells to tumor cells. The activity of bispecific proteins can also elicit immune responses due to inhibition of inhibitory checkpoint proteins or other immunosuppressive signals. The activity of bispecific proteins can also elicit an immune response due to activation of signals that elicit enhanced immune cell activity. In some embodiments, the bispecific protein is selected from ALT-801(CAS #1188450-53-4) and MEDI-565(AMG 211, BIIB-024, CAS # 1419574-83-6).

In one embodiment, the second compound is an EGFR inhibitor. EGFR inhibitors are compounds that prevent activation, upregulation, or overexpression of EGFR (which can be achieved in a variety of ways). Compounds that act on other proteins and also prevent activation, upregulation, or overexpression of EGFR are also considered EGFR inhibitors. EGFR upregulation or overexpression is caused by mutations in genes that produce uncontrolled cell division. EGFR upregulation or overexpression is associated with a variety of cancers, including but not limited to lung squamous cell carcinoma, anal carcinoma, glioblastoma, and epithelial tumors of the head and neck. In some embodiments, the EGFR inhibitor is selected from butkitabine (brigitinib), gefitinib (gefitinib), erlotinib (icotinib), neratinib (neratinib), afatinib (afatinib), dacomitinib (dacomitinib), cetuximab (cetuximab), erlotinib (erlotinib), flaveridine (flavididol), zalutumumab (zalutumumab), cetuximab (necitumumab), lidocaine (lidocaine), matuzumab (amatsumacumab), ocandib (osiritertinib), panitumumab (panitumumab), PD 393(CAS #194423-15-9), lapatinib (lapatinib), vandetanib (vatanib), rinpidotinib (huertinib), huertinib (ciclopin-max), and EGFR-vaegf.

In one embodiment, the second compound is a HER2 inhibitor. HER2 is also known as CD340, ERBB2 or HER 2/neu. HER2 is an oncogene that can activate a variety of cellular pathways, including the MAPK, PI3K/Akt, phospholipase C, PKC, and STAT pathways. Signal transduction by HER2 protein promotes cell proliferation and inhibits apoptosis. Inhibition of HER2 will decrease proliferation and increase apoptosis. HER2 inhibitors include small molecules, HER2 antagonists, inhibitory peptides, and anti-HER 2 antibodies. In some embodiments, the HER2 inhibitor is selected from ado-trastuzumab emtansine, trastuzumab (trastuzumab), and pertuzumab (pertuzumab).

In one embodiment, the second compound is an immunostimulatory therapeutic agent. An immunostimulatory therapeutic is any compound that acts as a stimulator of a stimulatory pathway or an inhibitor of an inhibitory pathway to modulate the activity of the immune system, such as an antibody, small molecule, biologic, or polysaccharide, which may differ from the previously described molecules and classes of molecules. The mechanism of action of these therapeutic agents may include activity as agonists, antagonists, allosteric effectors, enzymes, or any activity that results in an enhanced efficacy of the immune system against cancer. Immunostimulatory therapeutics may inhibit B7-H3, inhibit NKG2A, bind to phosphatidylserine, bind to CD27 to stimulate immune response resistance, antagonize the adenosine a2 receptor, or act by unknown mechanisms. In some embodiments, the immunostimulatory therapeutic agent is selected from vidapentan, varliumab, monelizumab, KAHR-102, BGB324(R-428, CAS #1037624-75-1), enoblizumab, liriluzumab, bavituximab (bavituximab), pidilizumab, BL-8040(CAS #664334-36-5), GDC-0919(NLG-919, RG607, CAS #1402836-58-1), IGN-311(CAS #1354846-06-2), elotuzumab (elotuzumab), bornauzumab (blinatumomab), salizumab, plerixafort (plerixafor), ganitumab, pexodartinib, trabeider, and galiniservib.

D. Combination of

The chromene compounds of formula (I) described herein are used in combination with a second compound known to be useful in the treatment or amelioration of an analogous disease, for example cancer. In combined administration, the second compound may be administered by the route of administration and in the usual dosage, simultaneously with or sequentially with the compound of formula (I). The chromene compound may be administered before or after the second compound. When the chromene compound of formula (I) is used simultaneously with the second compound, a pharmaceutical composition comprising the chromene compound of formula (I), the second compound and optionally one or more additional drugs may be used. Combination therapy also includes therapies in which the chromene compound of formula (I) and the second compound are administered on an overlapping schedule. The chromene compound of formula (I) when used in combination with the second compound may be used at a lower dosage than when the compound of formula (I) is used alone.

In one embodiment, the chromene compound of formula (I) is combined with a PD-1 inhibitor. In another embodiment, the chromene compound of formula (I) is deuterated and is combined with an inhibitor of PD-1. In another embodiment, one or more of compounds a01-a10 is combined with a PD-1 inhibitor. In another embodiment, the PD-1 inhibitor in combination with the deuterated chromene compound of formula (I) is nivolumab, pidilizumab, palbociclizumab, AMP-224, AMP-514, STI-A1110, TSR-043, AMP-514, or AUNP-12.

In another embodiment, the chromene compound of formula (I) is non-deuterated and is combined with an inhibitor of PD-1. In another embodiment, one or more of compounds B01-B11 is combined with a PD-1 inhibitor. In another embodiment, the PD-1 inhibitor in combination with the deuterated chromene compound of formula (I) is nivolumab, pidilizumab, palbociclizumab, AMP-224, AMP-514, STI-A1110, TSR-043, AMP-514, or AUNP-12.

In one embodiment, the chromene compound of formula (I) is combined with a PD-L1 inhibitor. In another embodiment, the chromene compound of formula (I) is deuterated and is combined with an inhibitor of PD-L1. In another embodiment, one or more of compounds a01-a10 is combined with a PD-L1 inhibitor. In another embodiment, the PD-L1 inhibitor in combination with the deuterated chromene compound of formula (I) is RG 7446, BMS-936559, MSB0010718C, STI-a1010, avizumab, atezolizumab or doxazozumab.

In another embodiment, the chromene compound of formula (I) is non-deuterated and is combined with an inhibitor of PD-L1. In another embodiment, one or more of compounds B01-B11 is combined with a PD-L1 inhibitor. In another embodiment, the PD-L1 inhibitor in combination with the deuterated chromene compound of formula (I) is RG 7446, BMS-936559, MSB0010718C, STI-a1010, avizumab, atezolizumab or doxazozumab.

In one embodiment, the chromene compound of formula (I) is combined with a CTLA-4 inhibitor. In another embodiment, the chromene compound of formula (I) is deuterated and is combined with an inhibitor of CTLA-4. In another embodiment, one or more of compounds a01-a10 is combined with a CTLA-4 inhibitor. In another embodiment, the CTLA-4 inhibitor in combination with the deuterated chromene compound of formula (I) is ipilimumab or tremelimumab.

In another embodiment, the chromene compound of formula (I) is non-deuterated and is combined with an inhibitor of CTLA-4. In another embodiment, one or more of compounds B01-B11 is combined with a CTLA-4 inhibitor. In another embodiment, the CTLA-4 inhibitor in combination with the deuterated chromene compound of formula (I) is ipilimumab or tremelimumab.

In one embodiment, the chromene compound of formula (I) is combined with an OX-40 agonist. In another embodiment, the chromene compound of formula (I) is deuterated and is combined with an OX-40 agonist. In another embodiment, one or more of compounds A01-A10 is combined with an OX-40 agonist. In another embodiment, the OX-40 agonist in combination with the deuterated chromene compound of formula (I) is an anti-OX 40, TIM3 antibody or imutune IMP 701.

In another embodiment, the chromene compound of formula (I) is non-deuterated, and is combined with an OX-40 agonist. In another embodiment, one or more of compounds B01-B11 is combined with an OX-40 agonist. In another embodiment, the OX-40 agonist in combination with the deuterated chromene compound of formula (I) is an anti-OX 40, TIM3 antibody or immuttoneimp 701.

In one embodiment, the chromene compound of formula (I) is combined with a CD137 agonist. In another embodiment, the chromene compound of formula (I) is deuterated, and is combined with a CD137 agonist. In another embodiment, one or more of compounds a01-a10 is combined with a CD137 agonist. In another embodiment, the CD137 agonist in combination with the deuterated chromene compound of formula (I) is urelumab or utomicilumab.

In another embodiment, the chromene compound of formula (I) is non-deuterated and is combined with a CD137 agonist. In another embodiment, one or more of compounds B01-B11 is combined with a CD137 agonist. In another embodiment, the CD137 agonist in combination with the deuterated chromene compound of formula (I) is urelumab or utomicilumab.

In one embodiment, the chromene compound of formula (I) is combined with a LAG-3 inhibitor. In another embodiment, the chromene compound of formula (I) is deuterated, and is combined with an inhibitor of LAG-3. In another embodiment, one or more of compounds a01-a10 is combined with a LAG-3 inhibitor. In another embodiment, the LAG-3 inhibitor in combination with the deuterated chromene compound of formula (I) is BMS-986016.

In another embodiment, the chromene compound of formula (I) is non-deuterated and is combined with an inhibitor of LAG-3. In another embodiment, one or more of compounds B01-B11 is combined with a LAG-3 inhibitor. In another embodiment, the LAG-3 inhibitor in combination with the deuterated chromene compound of formula (I) is BMS-986016.

In one embodiment, the chromene compound of formula (I) is combined with an IDO inhibitor. In another embodiment, the chromene compound of formula (I) is deuterated, and is combined with an inhibitor of IDO. In another embodiment, one or more of compounds a01-a10 is combined with an IDO inhibitor. In another embodiment, the IDO inhibitor in combination with the deuterated chromene compound of formula (I) is GDC-0919, indoximod, 1-methyl-D-tryptophan, NLG919, epacadostat, or norharman.

In another embodiment, the chromene compound of formula (I) is non-deuterated and is combined with an inhibitor of IDO. In another embodiment, one or more of compounds B01-B11 is combined with an IDO inhibitor. In another embodiment, the IDO inhibitor in combination with the deuterated chromene compound of formula (I) is GDC-0919, indoximod, 1-methyl-D-tryptophan, NLG919, epacadostat, or norharman.

In one embodiment, the chromene compound of formula (I) is combined with a bispecific protein. In another embodiment, the chromene compound of formula (I) is deuterated, and is combined with a bispecific protein. In another embodiment, one or more of compounds a01-a10 is combined with a bispecific protein. In another embodiment, the bispecific protein in combination with the deuterated chromene compound of formula (I) is ALT-801 or MEDI-565.

In another embodiment, the chromene compound of formula (I) is non-deuterated and is combined with a bispecific protein. In another embodiment, one or more of compounds B01-B11 is combined with a bispecific protein. In another embodiment, the bispecific protein in combination with the deuterated chromene compound of formula (I) is ALT-801 or MEDI-565.

In one embodiment, the chromene compound of formula (I) is combined with an EGFR inhibitor. In another embodiment, the chromene compound of formula (I) is deuterated, and is combined with an inhibitor of EGFR. In another embodiment, one or more of compounds a01-a10 is in combination with an EGFR inhibitor. In another embodiment, the EGFR inhibitor in combination with the deuterated chromene compound of formula (I) is bregtabine, gefitinib, erlotinib, lenatinib, afatinib, dacomitinib, cetuximab, erlotinib, fradaxin, zalutumumab, tolituzumab ozogamicin, lidocaine, matuzumab, oxitizumab, panitumumab, PD168393, lapatinib, vandetanib, ridopiumut, canertinib, HuMAX-EGFR or CimaVax-EGF.

In another embodiment, the chromene compound of formula (I) is non-deuterated and is combined with an inhibitor of EGFR. In another embodiment, one or more of compounds B01-B11 is combined with an EGFR inhibitor. In another embodiment, the EGFR inhibitor in combination with the deuterated chromene compound of formula (I) is bregtabine, gefitinib, erlotinib, lenatinib, afatinib, dacomitinib, cetuximab, erlotinib, fradaxin, zalutumumab, tolituzumab ozogamicin, lidocaine, matuzumab, oxitizumab, panitumumab, PD168393, lapatinib, vandetanib, ridopiumut, canertinib, HuMAX-EGFR or CimaVax-EGF.

In one embodiment, the chromene compound of formula (I) is combined with a HER2 inhibitor. In another embodiment, the chromene compound of formula (I) is deuterated, and is combined with an inhibitor of HER 2. In another embodiment, one or more of compounds a01-a10 is in combination with a HER2 inhibitor. In another embodiment, the HER2 inhibitor in combination with the deuterated chromene compound of formula (I) is ado-trastuzumab emtansine, trastuzumab, or pertuzumab.

In another embodiment, the chromene compound of formula (I) is non-deuterated and is combined with an inhibitor of HER 2. In another embodiment, one or more of compounds B01-B11 is in combination with a HER2 inhibitor. In another embodiment, the HER2 inhibitor in combination with the deuterated chromene compound of formula (I) is ado-trastuzumab emtansine, trastuzumab, or pertuzumab.

In one embodiment, the chromene compound of formula (I) is combined with an immunostimulatory therapeutic agent. In another embodiment, the chromene compound of formula (I) is deuterated, and is combined with an immunostimulatory therapeutic agent. In another embodiment, one or more of compounds a01-a10 is combined with an immunostimulatory therapeutic agent. In another embodiment, the immunostimulatory therapeutic agent in combination with the deuterated chromene compound of formula (I) is vidapentan, varliumab, monelizumab, KAHR-102, BGB324, enoblizumab, liriluzumab, bazedoximab, pidilizumab, BL-8040, GDC-0919, IGN-311, erlotinzumab, bornatemab, samalizumab, plerixafort, ganitumab, pexodartinib, trabedersen, and galuninsertib.

In another embodiment, the chromene compound of formula (I) is non-deuterated, and is combined with an immunostimulatory therapeutic agent. In another embodiment, one or more of compounds B01-B11 is combined with an immunostimulatory therapeutic agent. In another embodiment, the immunostimulatory therapeutic agent in combination with the deuterated chromene compound of formula (I) is vidapentan, varliumab, monelizumab, KAHR-102, BGB324, enoblizumab, liriluzumab, bazedoximab, pidilizumab, BL-8040, GDC-0919, IGN-311, erlotinzumab, bornatemab, samalizumab, plerixafort, ganitumab, pexodartinib, trabedersen, and galuninsertib.

In particular embodiments, compound a01 is combined with one of the following: erlotinib, palbocicluzumab, nivolumab, atezolizumab, ipilimumab, avizumab, druvalumab, trastuzumab, cetuximab, pertuzumab or panitumumab. In specific embodiments, compound a01 is combined with erlotinib.

The combinations in this application can be used with other conventional anti-inflammatory agents that are commercially available or under development, for example, drugs such as steroidal anti-inflammatory drugs, non-steroidal anti-inflammatory drugs, iNOS inhibitors, LTB4 receptor stimulators, and LTA4 hydrolase inhibitors, in order to enhance anti-inflammatory and analgesic effects; alternatively, it can be used in combination with antibiotics, alkylating drugs, antimetabolites, hormonal drugs, immunological drugs, interferon drugs, and some other drugs, in order to enhance the therapeutic or inhibitory effect on tumors.

E. Administration and dosage range

The compounds of formula (I), the second compounds, and combinations thereof of the present disclosure may be administered alone or as a pharmaceutical combination with pharmaceutically acceptable excipients for mammals (e.g., humans), for example, by oral, subcutaneous, intraperitoneal, intravenous, rectal, topical, ocular, pulmonary, nasal, and parenteral administration, based on standard pharmaceutical techniques.

In one embodiment, the chromene compound of formula (I) in the combination of the present disclosure is present in a therapeutically effective dose. In one embodiment, a therapeutically effective dose is an amount sufficient to cause a reduction in urinary PGE-M of at least 70% when compared to healthy controls or baseline standards. Urine PGE-M can be determined by conventional means such as enzyme-linked immunosorbent assay (ELISA) or mass spectrometry. The healthy control can be an individual who does not have cancer. The baseline standard may be obtained by determining the urinary PGE-M of the patient prior to initiation of the combined treatment with the chromene compound of formula (I) and the second compound. US 2012/0016002 describes a method of determining urine PGE-M of an individual and is incorporated herein by reference in its entirety.

In one embodiment, the dose of the chromene compound of formula (I) is from about 0.1 mg/kg/day to about 100 mg/kg/day. The dose may be administered as a single daily dose, or as two, three, four or more times per day, or in a sustained release form.

In one embodiment, the amount of the second compound is present in a therapeutically effective amount. In another embodiment, the therapeutically effective amount of the second compound is from about 0.01 mg/kg/day to about 250 mg/kg/day. In another embodiment, the therapeutically effective amount of the second compound is less when administered in combination with the compound of formula (I) than when administered alone. Specific therapeutically effective amounts of the second compound are disclosed in table 3.

Table 3: therapeutically effective dose of the second compound

F. Cancer treatment:

the combinations of the present disclosure are useful for treating cancer. In one embodiment, a method of treating cancer comprises administering to an individual in need thereof a therapeutically effective amount of a combination of a chromene compound of formula (I) and a second compound described herein. In particular embodiments, the cancer is selected from melanoma, non-small cell lung cancer, colorectal cancer, head and neck cancer, renal cell carcinoma, lymphoma, urothelial cancer, merkel cell carcinoma, pancreatic cancer, breast cancer, gastric cancer, intestinal cancer, endometrial cancer, hepatobiliary cancer, urinary tract cancer, brain cancer, skin cancer, glioblastoma, prostate cancer, and ovarian cancer. In particular embodiments, the cancer is colorectal cancer, gastric cancer, non-small cell lung cancer, breast cancer, pancreatic cancer, prostate cancer, or head and neck squamous cell carcinoma.

In another embodiment, the method further comprises determining expression of Major Histocompatibility Complex (MHC) class I in the cancer of the individual, and administering a combination of the chromene compound of formula (I) and a second compound when the cancer shows positive expression of MHC class I. MHC class I expression can be classified as high, low or negative, with high and low expression considered "positive" MHC class I expression. MHC class I expression can be quantified by Immunohistochemical (IHC) analysis or other clinical assays. The expression "high" and "low" can be determined by one of ordinary skill in the art. Soluble MHC class I polypeptide-related sequences A (sMICA), sMICB, soluble UL 16-binding protein (sULBP) -1, sULBP-2, sULBP-3, and sULBP-4 were measured using a custom multiplex bead array (R & D System). The bead-based assay was analyzed using the Luminex-based Bio-Plex system (BIO-RAD). For more information on the method of determining the level of soluble MHC class I polypeptide-associated chain a (sMICA) see Koguchi et al Cancer res.2015. The presence of MHCI type protein expression, as determined by standard methods by one skilled in the art (e.g., a clinical pathologist), is a predictor of positive response to the claimed combination therapy. See Simpson et al Gut 2010.

In another embodiment, the method further comprises determining the expression of PD-L1 in the cancer of the individual, and administering to the individual a combination of the chromene compound of formula (I) and a second compound when the cancer shows positive expression of PD-L1. PD-L1 can be determined by methods conventional in the art (e.g., IHC assay). In one embodiment, a tumor may be considered positive for PD-L1 expression if stained for 150% or more of the tumor cells of PD-L.

In another embodiment, the method further comprises determining the level of intratumoral T cells in the cancer of the individual, and administering to the individual a combination of the chromene compound of formula (I) and the second compound when the cancer shows an elevated intratumoral T cell level. Greater than 15T cells/mm in colorectal cancer, assessed by the standard method of IHC staining, according to Simpson et al, Gut 2010, in colorectal cancer²Are considered to be high or elevated intratumoral T cell levels. According to Dieci Annals of Oncology 2015, in breast cancer, if there are greater than or equal to 50% intratumoral T cells (It-TIL) or stromal T cells (Str-TIL) according to the method disclosed in Salgado et al Ann Oncol (2015), the situation is defined as elevated or high-containing intratumoral T cells, also known as Tumor Infiltrating Lymphocytes (TIL).

In another embodiment, the method further comprises determining the subject's urine PGE-M level, and administering to the subject a combination of the chromene compound of formula (I) and the second compound when the urine PGE-M level is elevated. Urine PGE-M levels are considered "elevated" when they are at least 1.5 times the Upper Limit of Normal (ULN). For males, elevated urinary PGE-M levels will be >15ng/mg creatinine (ULN 10ng/mg creatinine). For women, elevated urinary PGE-M levels will be >9ng/mg creatinine (ULN 6ng/mg creatinine). In another embodiment, the urinary PGE-M level is measured in an individual having a cancer selected from colorectal cancer, non-small cell lung cancer, breast cancer, gastric cancer, pancreatic cancer, prostate cancer, or head and neck squamous cell carcinoma. In another embodiment, the colorectal cancer, non-small cell lung cancer, breast cancer, gastric cancer, pancreatic cancer, prostate cancer, or head and neck squamous cell carcinoma is at stage III or stage IV.

In another embodiment, the method further comprises determining the level of microsatellite instability (MSI) in the individual and administering a combination of the chromene compound of formula (I) and a second compound when the MSI is high, low or stable. MSI can be determined by means conventional in the art, e.g., by PCR-based assays for specific DNA repeats or IHC assays for mismatch repair (MMR) proteins (see, e.g., Vilar et al, nat. rev. Clin. oncol, 2010; Bupathi et al, j. gastrointest. oncol, 2016; Dudley et al, Clin Cancer res.2016; Sinicrope et al, Clin. gastroentol. hepatol. 2016; and Kautto et al, Oncotarget, 2016). In one embodiment, high levels of MSI may be defined as instability at two or more loci, or instability at > 30% of loci in a labeled large panel (panel); low levels of MSI can be defined as instability of one locus, or instability of 10-30% of loci in a larger panel; and microsatellite stability can be defined as no instability at any locus, or instability at < 10% of loci in a larger panel. In another embodiment, the method further comprises determining microsatellite instability when the cancer is colorectal cancer, gastric cancer, endometrial cancer, ovarian cancer, cancer of the liver and biliary tract, cancer of the urinary tract, cancer of the brain, or skin cancer.

In another embodiment, the method further comprises determining CD8 according to standard flow cytometry methods^+/The proportion of FOXP3 expressing cells, and when CD8⁺/FOXP3 ratio>A combination of a chromene compound of formula (I) and a second compound is administered at 1 (which is reported to be predictive of excellent clinical outcome for ovarian and urothelial cancers, Preston et al PLoS one.2013 and Baras et al Oncoimmunology 2016).

In a specific embodiment, the method of treating lung cancer comprises administering a combination of compound a01 and erlotinib.

In particular embodiments, the method of treating colorectal cancer comprises administering a combination of compound a01 and palivizumab.

In a specific embodiment, the method of treating melanoma comprises administering a combination of compound a01 and palivizumab.

In a specific embodiment, the method of treating colorectal cancer comprises administering a combination of compound a01 and atezolizumab.

In a specific embodiment, the method of treating lung cancer comprises administering a combination of compound a01 and atezolizumab.

G. The pharmaceutical composition comprises:

the pharmaceutical composition containing the chromene compound of formula (I), the second compound, or a combination thereof may be in a form suitable for oral administration, for example, a tablet, a troche, an aqueous or oily suspension, a dispersible powder or dispersible granules, an emulsion, a hard or soft capsule, or a syrup or elixir. Compositions for oral administration may be prepared according to any method known to the art for the manufacture of pharmaceutical compositions, and such compositions may contain one or more excipients or agents selected from the group consisting of sweetening agents, flavoring agents, coloring agents and preserving agents in order to provide pharmaceutically elegant and palatable preparations. Tablets contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients which are suitable for the manufacture of tablets. These excipients may be, for example, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, for example, microcrystalline cellulose, croscarmellose sodium, corn starch or alginic acid; binding agents, for example starch, gelatin, polyvinylpyrrolidone or acacia; and lubricating agents, for example, magnesium stearate, stearic acid or talc. The tablets may be uncoated or they may be coated by known techniques to mask the unpleasant taste of the drug or to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a water soluble taste masking material such as hydroxypropylmethyl cellulose or hydroxypropyl cellulose may be used, or a time delay material such as ethyl cellulose or cellulose acetate butyrate may be used.

The dosage of the tablet may be 0.1mg/tab, 0.2mg/tab, 0.25mg/tab, 0.5mg/tab, 1mg/tab, 2mg/tab, 5mg/tab, 10mg/tab, 25mg/tab, 50mg/tab, 100mg/tab and 250 mg/tab. Other forms of dosage, such as capsules, may be similarly referred to.

Formulations for oral use may also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin; or as soft gelatin capsules wherein the active ingredient is mixed with a water-soluble carrier, for example polyethylene glycol, or an oil medium, for example peanut oil, liquid paraffin, or olive oil.

Aqueous suspensions contain the active material in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients include suspending agents, for example, sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia; dispersing or wetting agents may be a naturally-occurring phosphatide, for example lecithin, or condensation products of an alkylene oxide with fatty acids, for example polyoxyethylene stearate, or condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example polyethylene sorbitan monooleate. The aqueous suspension may also contain one or more preservatives, for example ethyl or n-propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents, such as sucrose, saccharin or aspartame.

oily suspensions may be formulated by suspending the active ingredient in a vegetable oil, for example arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin.

Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water provide the active ingredient in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents have been exemplified by those mentioned above. Additional excipients, for example sweetening, flavoring and coloring agents, may also be present. These compositions may be preserved by the addition of an antioxidant such as ascorbic acid.

The pharmaceutical compositions of the present disclosure may also be in the form of oil-in-water emulsions. The oily phase may be a vegetable oil, for example olive oil or arachis oil, or a mineral oil, for example liquid paraffin, or a mixture of such oils. Suitable emulsifiers may be naturally occurring phospholipids, such as soy lecithin; and esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan monooleate; and condensation products of the said partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate. The emulsion may also contain sweeteners, flavoring agents, preservatives and antioxidants.

Syrups and elixirs may be formulated with sweetening agents, for example glycerol, propylene glycol, sorbitol or sucrose. Such formulations may also contain a demulcent, a preservative, a flavoring agent, a coloring agent and an antioxidant.

The pharmaceutical compositions may be in the form of a sterile injectable aqueous solution. Among the acceptable carriers and solvents, water, ringer's solution and isotonic sodium chloride solution may be used.

The sterile injectable preparation may also be a sterile injectable oil-in-water microemulsion in which the active ingredient is dissolved in an oil phase. For example, the active ingredient may be first dissolved in a mixture of soybean oil and lecithin. The oil solution is then introduced into a mixture of water and glycerol and processed to form a microemulsion.

Injectable solutions or microemulsions may be introduced into the bloodstream of a patient by means of a local bolus injection. Alternatively, it may be advantageous to apply the solution or microemulsion in a manner that maintains a constant circulating concentration of the compound of the present invention. To maintain such a constant concentration, a continuous intravenous delivery device may be used. One embodiment of such a device is Deltec CADD-PLUS^TMModel 5400 intravenous pump.

The pharmaceutical compositions may be in the form of sterile injectable aqueous or oleaginous suspensions for intramuscular and subcutaneous administration. Such suspensions are formulated according to known techniques using those suitable dispersing or wetting agents and suspending agents which have been mentioned above. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent, for example as a solution in 1, 3-butanediol. In addition, nonvolatile oils are commonly used as solvents or suspending media. For this purpose, any bland fixed oil may be employed including synthetic mono-or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables.

The chromene compound of formula (I), the second compound, or a combination thereof may also be administered in the form of a suppository for rectal administration of the drug. These compositions can be prepared by mixing the drug with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Such materials include cocoa butter, glycerogelatin, hydrogenated vegetable oils, mixtures of polyethylene glycols of various molecular weights, and fatty acid esters of polyethylene glycols.

For topical use, creams, ointments, jellies, solutions or suspensions, etc., containing the chromene compound of formula (I), the second compound, or a combination thereof are used (for this use, topical use shall include mouthwashes and rinses).

The chromene compound of formula (I), the second compound, and combinations thereof of the present disclosure may be administered in intranasal form by topical use of suitable intranasal vehicles and delivery devices, or in transdermal routes by use of those transdermal patch forms well known to those of ordinary skill in the art. For administration in the form of a transdermal delivery system, of course, the dosage administration will be continuous rather than intermittent throughout the dosage regimen. The compounds and combinations of the present disclosure may also be delivered as suppositories using the following bases; such as cocoa butter, glycerogelatin, hydrogenated vegetable oils, mixtures of polyethylene glycols of various molecular weights, and fatty acid esters of polyethylene glycols.

When a compound of the present disclosure is administered to a human subject, the daily dosage will typically be determined by the prescribing physician, and the dosage will typically vary with the age, weight, sex, and response of the individual patient, as well as the severity of the patient's symptoms.

H. Metabolites and prodrugs:

combinations of the present disclosure also include combinations of the chromene compound of formula (I) and metabolites and/or prodrugs of the second compound. In one embodiment, the combination comprises a metabolite or prodrug of a chromene compound and a second compound. In another embodiment, the combination comprises a chromene compound and a metabolite or prodrug of the second compound. In another embodiment, the combination comprises a metabolite or prodrug of the chromene compound and a metabolite or prodrug of the second compound.

Examples

Example 1: combined anti-tumor effects in colon cell carcinoma

This study evaluated the effect of (S) -6-bromo-8-trideuteromethyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid (compound a01), a selective cyclooxygenase 2(COX-2) inhibitor, in combination with erlotinib on tumor growth in an HT-29 xenografted murine colon cancer model.

HT-29 cells were cultured in medium for one week. After digestion, cells were centrifuged at 800-1500rpm for 3-5 minutes. Cells were washed with PBS and centrifuged again under the same conditions. Cells were then suspended in PBS until the final concentration was 12.5x10⁷Individual cells/mL. mu.L of HT-29 cell suspension was injected subcutaneously into the left forelimb axilla (2.5X 10)⁶One HT-29 cell/mouse).

Implantation of 2.5 × 10 to 48 CB17SCID male mice⁶HT29 cells. Using calipers and the equation V π × a b²The growth of the tumors was calculated for each animal. Once the tumor reaches about 75mm³The mice were then randomly assigned to one of 6 treatment groups: vehicle, 1mg/kg Compound A01, 1mg/kg Compound A01+50mg/kg erlotinib, 10mg/kg Compound A01, 10mg/kg Compound A01+50mg/kg erlotinib, 50mg/kg erlotinib. Compound A01 was dissolved in 2% DMSO, 4% ethanol, 4% castor oil, and 90% ddH₂And (4) in O.

Compound a01 and erlotinib were administered daily by oral gavage, starting immediately after randomization and lasting for 22 days. Mouse weight and tumor volume measurements were performed on days 0, 3, 6, 9, 12, 15, 18, 21, and 22 after randomization. Tumor tissue was collected from each animal on day 24 to determine compound a01, erlotinib, and PGE2 levels.

All animals developed tumors. There were no differences in body weight between treatment groups during the study (Table 6)

Table 6: body weight (g)

The combination of 1mg/kg compound a01+50mg/kg erlotinib slowed tumor growth by 66% when compared to vehicle-treated mice. The combination of 10mg/kg compound a01+50mg/kg erlotinib slowed tumor growth by 60% when compared to vehicle-treated mice. Compound a01 alone produced 51% inhibition of tumor growth compared to vehicle, whereas erlotinib alone produced 38% inhibition of tumor growth when compared to vehicle (table 7, figure 1).

Table 7: tumor volume (mm) of treatment group³)

The oral dose of erlotinib affected the plasma concentrations of compound a01 at 2 and 6 hours post-administration (table 8).

Table 8: plasma concentrations of Compound A01 and erlotinib (mean. + -. SD,. mu.g/L)

Addition of erlotinib increased the intratumoral level of compound a01 by 1.5-fold when compared to the amount of intratumoral compound a01 in the absence of erlotinib alone (table 9).

Table 9: intratumoral concentrations of Compound A01 and erlotinib (mean. + -. SD, ug/g)

Treatment group	Compound A01	Erlotinib
			Media	0	0
Compound A011 mg/kg	1478.5±652.1
			Compound A011 mg/kg + erlotinib-50 mg/kg	2243.5±887.1	13225±3017.7
Compound A0110 mg/kg	9720±2758.7
			Compound A0110 mg/kg + erlotinib-50 mg/kg	14745±4758	7800±3299
Erlotinib-50 mg/kg		<80

Compound a01 alone or in combination with erlotinib inhibited PGE2 levels within tumors by 66-100% (table 10).

Table 10: intratumoral concentration of PGE2 (mean. + -. SD, ug/g)

Treatment group	PGE2
		Media	246.7±104.3
Compound A011 mg/kg	14.01±30.3
		Compound A011 mg/kg + erlotinib-50 mg/kg	85.3±110.9
Compound A0110 mg/kg	0
		Compound A0110 mg/kg + erlotinib-50 mg/kg	12.8±18.3
Erlotinib-50 mg/kg	160.6±89.5

For slowing tumor growth, Compound A01 alone was superior to 50mg/kg erlotinib. However, the combination of compound a01+ erlotinib showed the greatest inhibition of tumor growth compared to each individual treatment when compared to vehicle. When combined with erlotinib, there was little difference between compound a01 administered at 1mg/kg or 10 mg/kg.

Example 2: combined anti-tumor effects in colon cell carcinoma

This experiment evaluated the effect of compound a01 in combination with erlotinib on tumor growth in a CT26 xenografted murine colon cancer model.

Ct26.wt cells were cultured under standard conditions. The cells were harvested with trypsin and washed with PBS. Will 10⁵Individual cells were injected in a total volume of 100 μ Ι _ into the right flank of 6-week-old female BALB/c mice. Tumor size was quantified as the average of the longest diameter and its perpendicular diameter. From day 0 onwards, mice received celecoxib (celecoxib), compound a01, or vehicle (10% DMSO/50% PEG 400/40% water) orally (per o.s.) daily. anti-PD-1 monoclonal antibody (clone RMP1-14, BioXCell) was administered intraperitoneally at 200 μ g/mouse from day 5 to day 9 (when the mean tumor diameter was about 5 mm) after tumor cell inoculation, followed by a maximum of six injections every 3 to 4 days.

mice receiving combination treatment with a COX-2 inhibitor (compound A01 or celecoxib) and an anti-PD 1 antibody completely rejected tumors up to 50 days post-vaccination, in contrast, monotherapy (celecoxib alone, compound A01, or anti-PD 1 antibody) did not prevent tumor growth (FIG. 2), no treatment regimen caused weight loss₂Intratumoral levels of (a).

Example 3: combining effects on immune response to colon cell carcinoma

Female BALB/c mice were inoculated with 10 as in example 2⁵CT26 colorectal cancer cells. COX-2 inhibitor (celecoxib or compound a01) or vehicle was administered orally daily. anti-PD 1 antibody (200 μ g/mouse) was administered intraperitoneally on days 9 and 14 post-inoculation. Tumors were analyzed on day 16 (7 days after the first anti-PD 1 administration).

On day 16, there was no difference in tumor size or weight between groups. Monotherapy or combination therapy with compound a01 had CD45 compared to vehicle-treated mice⁺Cells (leukocytes), CD3⁺、CD8⁺、CD8⁺IFNγ⁺、CD4⁺IFNγ⁺、CD8⁺TNFα⁺And CD4⁺TNFα⁺Increased penetration of cells into the tumor. Administration of anti-PD 1 antibody alone or in combination with a COX-2 inhibitor increases intratumoral CD4⁺Foxp3⁺The number of cells. Treatment with Compound A01 alone increased CD8⁺T cells with Foxp3⁺Proportion of T cells. Treatment of celecoxib alone did not increase CD8⁺T cells with Foxp3⁺Proportion of T cells. For GR-1⁺Bone marrow-derived suppressor cells (MDSCs) had no apparent effect. Combination therapy increased splenic CD4⁺IFNγ⁺The number of cells.

Example 4: effect of the combination of chromene and anti-PD-L1 antibody on tumor growth and immune response

This experiment evaluated the effect of immune response to treatment with compound a01 alone and in combination with an anti-PD-L1 antibody in a murine colon cancer model.

CT26 murine colon cancer cells were prepared as described in example 2. Female Envigo BALB/c (BALB/cAnNHsd) was inoculated subcutaneously at high axillary elevations (just below the forelimb) with 5X10 suspended in PBS (200. mu.L)⁵And CT26 cells. Mice were divided into seven groups: vehicle (group 1), antibody control (rat IgG2b isotype) (group 2), anti-mPD-L1 (clone 10F9G2) (group 3), Compound A01 (group 4), Compound A01+Antibody control (group 5), Compound A01+ anti-mPD 1-L1 (group 6), and 1mg/kg Compound A01+ anti-mPD-L1 (group 7). The antibodies were administered by intraperitoneal injection on days 3, 6, 9, 12 and 16 after inoculation. Compound a01 was administered orally daily. Mice were euthanized 2 hours after the final dose on day 16 and samples were prepared for testing.

Treatment with compound a01 slowed tumor growth compared to vehicle and isotype control, or treatment with anti-mPD-L1 slowed tumor growth (all doses were 10mg/kg unless indicated).

Table 11: effect of Compound A01 and anti-mPD-L1 on tumor growth (mm)³)

1mg/kg Compound A01

After euthanasia, tumors were removed and processed to recover viable cells. The red blood cells were removed. Cells are stained for surface markers and, where appropriate, permeabilized for intracellular staining. Cells were washed and suspended in flow cytometry staining buffer and analyzed by flow cytometry.

Table 12 shows the effect of Compound A01(10mg/kg, unless otherwise indicated) and anti-mPD-L1 (10mg/kg) on the immune response. Compound A01 alone and in combination with anti-mPD-L1 increased intratumoral CD4⁺CD8^-Percentage of T cells (for total CD 45)⁺For cells). Both Compound A01 and anti-mPD-L1 increased intratumoral CD8⁺CD4^-Percentage of T cells (for total CD 45)⁺For cells). Compound A01 reduced Ki67⁺CD8⁺And PD-1⁺CD8⁺Level of T cells (for total CD 8)⁺CD4^-For T cells). Treatment for Treg (CD 25)⁺Foxp3⁺) And MDSC (CD11 b)⁺Ly6G⁺) As a total of CD45⁺Of cellsPercentage) has the least effect. Combined treatment improved CD8⁺Ratio of T cells to tregs. FIG. 3 shows CD8⁺The percentage of cells correlates with tumor regression. FIG. 4 shows a CD8⁺Statistical differences between groups.

Table 12: effect of Compound A01 and anti-mPD-L1 on immune response in tumors

1mg/kg Compound A01

Example 5: effect of Compound A01 on HT-29 tumor growth and intratumoral PGE2

This study compared the effect of compound a01 and celecoxib on the effect of tumor PGE-2 in a HT-29 xenograft murine colon cancer model.

HT-29 cells were prepared and implanted by standard methods using the same procedure as in example 1.

Implantation of 2.5X10 into 42 CB17SCID mice⁶And HT-29 cells. Once the tumor reached 75mm³The mice were then randomly assigned to one of six treatment groups: vehicle, 10mg/kg celecoxib, 0.1mg/kg Compound A01, 0.3mg/kg Compound A01, 1mg/kg Compound A01, 3mg/kg Compound A01, 10mg/kg Compound A01. Compound a01 was dissolved in 2% DMSO, 4% ethanol, 4% castor oil, and 90% ddH 2O.

Compound a01 and celecoxib were administered daily by oral gavage, starting immediately after randomization and lasting 22 days. Mouse weight and tumor volume measurements were performed on days 0, 3, 6, 9, 12, 15, 18, 21, and 22 after randomization. After 22 days of dosing, blood was collected from each animal at 2 and 6 hours post-dose. The concentration of compound a01 in plasma was measured. Tumor tissue was collected from each animal on day 24 to determine compound a01, celecoxib, and PGE2 levels.

Table 13 shows the effect of 10mg/kg celecoxib or various doses of compound a01 on tumor volume. Table 14 shows the mean intratumoral PGE-2 plasma concentrations after treatment with Compound A01 or celecoxib.

Table 13: tumor volume (mm) of treatment group³)

Table 14: tumor PGE-2 plasma concentrations in treatment groups

Example 6: effect of Compound A01 on CT26 tumor growth and intratumoral PGE2

This experiment evaluated the efficacy of compound a01 compared to celecoxib, as well as their effect on prostaglandin E2(PGE2) levels and T cell inhibition in Balc/c female mice with ct26.wt murine colon cancer. In addition, analysis of the concentration of compound a01 in blood and tumors was performed.

CT26 murine colon cancer cells were prepared as described in example 2. Female Harlan Balb/c mice (BALB/cAnNHsd) were inoculated subcutaneously at high axillary elevations (just below the forelimb) with 5X10 cells suspended in PBS (200. mu.L)⁵And CT26 cells. When the mean tumor burden of all animals was about 79mm on day 8³All mice were classified into study groups based on caliper estimation of tumor burden (group mean range, 75-83 mm)³). The mice were divided into 10 groups: vehicle control (group 1), 30mg/kg CompoundSubstance A01 (group 2), 10mg/kg Compound A01 (group 3), 3mg/kg Compound A01 (group 4), 1mg/kg Compound A01 (group 5), 0.3mg/kg Compound A01 (group 6), 30mg/kg celecoxib (group 7), and 10mg/kg celecoxib (group 8).

Table 15 shows the effect of 10mg/kg celecoxib or various doses of compound a01 on tumor volume. Table 16 shows the mean intratumoral PGE-2 plasma concentrations after treatment with Compound A01 or celecoxib.

Table 15: CT26 tumor volume (mm) of treatment group³)

Table 16: CT26 tumor PGE-2 plasma concentration (ng/g) of treatment group

Treatment group	Average PGE-2 concentration	Standard deviation of
			Medium (group 1)	1180	610
Compound A0130 mg/kg (group 2)	150	60
			Compound A011 mg/kg (group 5)	278	141
Compound A010.3mg/kg (group 6)	670	300
			Celecoxib 10mg/kg (group 8)	1140	690

Example 7: effect of Compound A01 and anti-PD-L1 on CT26 tumor growth and intratumoral PGE2

This experiment evaluated the effect of treatment with compound a01 alone or in combination with an anti-PD-L1 antibody on PGE-2 plasma concentrations in a murine colon cancer model.

CT26 murine colon cancer cells were prepared as described in example 4. Female EnvigobaLB/c (BALB/cAnNHsd) mice were inoculated with 5X10 cells as described in example 4⁵And CT26 cells. The mice were then divided into control and treatment groups, with compound a01 treated either alone or in combination with anti-PD-L1 antibody. Tumor size and tumor PGE-2 plasma concentration were determined.

The embodiments mentioned above are merely illustrative of various aspects of the disclosure. These embodiments should not be considered as limiting the patent in any way. It should be noted that various changes and modifications can be made by those skilled in the art without departing from the concept of the present invention, and those changes and modifications should fall within the scope of the present invention. Accordingly, the scope of the invention is limited by the claims.

All documents mentioned are incorporated herein by reference as if written herein. When introducing elements of the present disclosure or the exemplary embodiments thereof, the articles "a", "an", "the" and "the" mean that there are one or more of the elements. The terms "comprising," "including," and "having" are intended to be inclusive and mean that there may be additional elements other than the listed elements. While the invention has been described with respect to specific embodiments, the details of these embodiments are not to be construed as limiting.

Claims (37)

1. A combination, comprising:

a compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof, and a second compound,

wherein M is selected from H and alkyl;

z is selected from CF₃、CF₂H and C₂F₅；

Each R is¹、R²、R³And R⁴Independently selected from the group consisting of H, alkyl, aralkyl, deuterated alkyl, deuterated aralkyl, deuterated alkoxy, deuterated cycloalkyl, deuteron, deuteroaryloxy, deuteroheteroaryloxy, deuteroarylalkoxy, deuteroheteroarylalkoxy, deuterohaloalkoxy, deuteroamino, deuterosulfonamido, sulfonamido, cycloalkyl, cycloalkenyl, halogen, haloalkyl, alkoxy, haloalkoxy, alkylthio, haloalkylthio, pentafluorosulfanyl, hydroxyalkyl, trialkylsilyl, alkynyl, and alkenyl; and

wherein the second compound is selected from the group consisting of a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, an OX-40 agonist, a CD137 agonist, a LAG-3 inhibitor, an IDO inhibitor, a bispecific protein, an EGFR inhibitor, a HER2 inhibitor, and an immunostimulatory therapeutic.

2. A combination according to claim 1, wherein R of formula (I)¹、R²、R³And R⁴At least one of which is selected from the group consisting of deuterated, deuterated alkyl, and deuterated cycloalkyl.

3. A combination according to claim 2 wherein R is²Is not H.

4. The combination of claim 3, wherein the compound of formula (I) is selected from:

(S) -6, 8-di-trideuteromethyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -6-bromo-8-trideuteromethyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -8-pentadeuterated ethyl-6- (trifluoromethoxy) -2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -6-chloro-8-trideuteromethyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -6-chloro-5, 7-di-trideuteromethyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -6-bromo-5, 7-di-trideuteromethyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -6-trideuteromethyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -8-chloro-6-trideuteromethyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -8-Trideuteromethyl-6- (pentafluorosulfanyl) -2- (trifluoromethyl) -2H-chromene-3-carboxylic acid, and

(S) -8-trideuteromethyl-6- (trifluoromethoxy) -2- (trifluoromethyl) -2H-chromene-3-carboxylic acid.

5. The combination of claim 2, wherein the second compound is:

PD-1 inhibitors selected from the group consisting of nivolumab, pidilizumab, pabollizumab, AMP-224, AMP-514, STI-A1110, TSR-043, AMP-514, and AUNP-12;

a PD-L1 inhibitor selected from RG 7446, BMS-936559, MSB0010718C, STI-A1010, avizumab, atlizumab and dulvacizumab;

a CTLA-4 inhibitor selected from ipilimumab or tremelimumab;

an OX-40 agonist selected from anti-OX 40, TIM3 antibody and Immuture IMP 701;

a CD137 agonist selected from urelumab and utomicilumab;

LAG-3 inhibitor BMS-986016;

an IDO inhibitor selected from GDC-0919, indoximod, 1-methyl-D-tryptophan, NLG919, epacadostat, and norharpagne;

an EGFR inhibitor selected from butitabine, gefitinib, erlotinib, neratinib, afatinib, dacomitinib, cetuximab, erlotinib, fradaxin, zalutumab, tolituzumab, lidocaine, matuzumab, oxitizumab, panitumumab, PD168393, lapatinib, vandetanib, ridapepimut, canertinib, HuMAX-EGFR and CimaVax-EGF;

a HER2 inhibitor selected from the group consisting of ado-trastuzumab emtansine, trastuzumab, and pertuzumab; and

an immunostimulatory therapeutic agent selected from vidapentan, varliumab, monelizumab, KAHR-102, BGB324, enoblizumab, lirilumab, bazedoximab, pidilizumab, BL-8040, GDC-0919, IGN-311, erlotinzumab, bornauzumab, samalizumab, plerixafu, ganitumab, pexodartinib, trabedersen, and galuninisertib.

6. The combination of claim 4, wherein the second compound is:

PD-1 inhibitors selected from the group consisting of nivolumab, pidilizumab, pabollizumab, AMP-224, AMP-514, STI-A1110, TSR-043, AMP-514, and AUNP-12;

a PD-L1 inhibitor selected from RG 7446, BMS-936559, MSB0010718C, and STI-A1010, avilamumab, atlizumab, and dulvacizumab;

a CTLA-4 inhibitor selected from ipilimumab or tremelimumab;

an OX-40 agonist selected from anti-OX 40, TIM3 antibody and Immuture IMP 701;

a CD137 agonist selected from urelumab and utomicilumab;

LAG-3 inhibitor BMS-986016;

an IDO inhibitor selected from GDC-0919, indoximod, 1-methyl-D-tryptophan, NLG919, epacadostat, and norharpagne;

a bispecific protein selected from ALT-801 and MEDI-565;

an EGFR inhibitor selected from butitabine, gefitinib, erlotinib, neratinib, afatinib, dacomitinib, cetuximab, erlotinib, fradaxin, zalutumab, tolituzumab, lidocaine, matuzumab, oxitizumab, panitumumab, PD168393, lapatinib, vandetanib, ridapepimut, canertinib, HuMAX-EGFR and CimaVax-EGF;

a HER2 inhibitor selected from the group consisting of ado-trastuzumab emtansine, trastuzumab, and pertuzumab; and

an immunostimulatory therapeutic agent selected from vidapentan, varliumab, monelizumab, KAHR-102, BGB324, enoblizumab, lirilumab, bazedoximab, pidilizumab, BL-8040, GDC-0919, IGN-311, erlotinzumab, bornauzumab, samalizumab, plerixafor, ganitumab, pexodartinib, trabedersen and galuninisertib.

7. The combination of claim 6, wherein the second compound is selected from erlotinib, palbociclumab, nivolumab, atuzumab, ipilimumab, avizumab, dovacizumab, trastuzumab, cetuximab, pertuzumab, and panitumumab.

8. The combination of claim 6, wherein the compound of formula (I) is (S) -6-bromo-8-trideuteromethyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid and the second compound is selected from erlotinib, palbociclumab, nivolumab, altuzumab, ipilimumab, avizumab, dutuzumab, trastuzumab, cetuximab, pertuzumab, and panitumumab.

9. The combination of claim 8, wherein the second compound is erlotinib, cetuximab, trastuzumab, or pertuzumab.

10. The combination of claim 8, wherein the compound of formula (I) is (S) -6-bromo-8-trideuteromethyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid and the second compound is pabulizumab, nivolumab, altuzumab, ipilimumab, doxitumumab, or avizumab.

11. The combination of claim 9, wherein the second compound is erlotinib.

12. The combination of claim 10, wherein the second compound is palivizumab, nivolumab, or atuzumab.

13. A combination according to claim 1 wherein each R of formula (I)¹、R²、R³And R⁴Independently selected from the group consisting of H, alkyl, aralkyl, cycloalkyl, cycloalkenyl, halo, haloalkyl, alkoxy, haloalkoxy, alkylthio, haloalkylthio, pentafluorosulfanyl, hydroxyalkyl, trialkylsilyl, alkynyl and alkenyl.

14. The combination of claim 13 of claim 1, wherein R¹Is H, and R²Selected from halogen, haloalkoxy and pentafluorosulfanyl, and R⁴Selected from H, alkyl, alkenyl, alkynyl and halogen.

15. The combination of claim 14, wherein the compound of formula (I) is selected from:

(S) -6, 8-dichloro-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -6-chloro-8-methyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -6-bromo-8-methyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -6, 8-dimethyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -8-methyl-6- (trifluoromethoxy) -2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -8-ethyl-6- (trifluoromethoxy) -2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -6-chloro-5, 7-dimethyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -7- (tert-butyl) -6-chloro-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -6-pentafluorosulfanyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -6-pentafluorosulfanyl-8-methyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid, and

(S) -6-pentafluorosulfanyl-8-ethyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid.

(S) -6-pentafluorosulfanyl-8-ethyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid.

16. The combination of claim 13, wherein the second compound is:

PD-1 inhibitors selected from the group consisting of nivolumab, pidilizumab, pabollizumab, AMP-224, AMP-514, STI-A1110, TSR-043, AMP-514, and AUNP-12;

a PD-L1 inhibitor selected from RG 7446, BMS-936559, MSB0010718C, STI-A1010, avizumab, atlizumab and dulvacizumab;

a CTLA-4 inhibitor selected from ipilimumab or tremelimumab;

an OX-40 agonist selected from anti-OX 40, TIM3 antibody and Immuture IMP 701;

a CD137 agonist selected from urelumab and utomicilumab;

LAG-3 inhibitor BMS-986016;

an IDO inhibitor selected from GDC-0919, indoximod, 1-methyl-D-tryptophan, NLG919, epacadostat, and norharpagne;

a bispecific protein selected from ALT-801 and MEDI-565;

an EGFR inhibitor selected from butitabine, gefitinib, erlotinib, neratinib, afatinib, dacomitinib, cetuximab, erlotinib, fradaxin, zalutumab, tolituzumab, lidocaine, matuzumab, oxitizumab, panitumumab, PD168393, lapatinib, vandetanib, ridapepimut, canertinib, HuMAX-EGFR and CimaVax-EGF;

a HER2 inhibitor selected from the group consisting of ado-trastuzumab emtansine, trastuzumab, and pertuzumab; and

an immunostimulatory therapeutic agent selected from vidapentan, varliumab, monelizumab, KAHR-102, BGB324, enoblizumab, lirilumab, bazedoximab, pidilizumab, BL-8040, GDC-0919, IGN-311, erlotinzumab, bornauzumab, samalizumab, plerixafor, ganitumab, pexodartinib, trabedersen and galuninisertib.

17. The combination of claim 15, wherein the second compound is:

PD-1 inhibitors selected from the group consisting of nivolumab, pidilizumab, pabollizumab, AMP-224, AMP-514, STI-A1110, TSR-043, AMP-514, and AUNP-12;

a PD-L1 inhibitor selected from RG 7446, BMS-936559, MSB0010718C, STI-A1010, avizumab, atlizumab and dulvacizumab;

a CTLA-4 inhibitor selected from ipilimumab or tremelimumab;

an OX-40 agonist selected from anti-OX 40, TIM3 antibody and Immuture IMP 701;

a CD137 agonist selected from urelumab and utomicilumab;

LAG-3 inhibitor BMS-986016;

an IDO inhibitor selected from GDC-0919, indoximod, 1-methyl-D-tryptophan, NLG919, epacadostat, and norharpagne;

a bispecific protein selected from ALT-801 and MEDI-565;

an EGFR inhibitor selected from butitabine, gefitinib, erlotinib, neratinib, afatinib, dacomitinib, cetuximab, erlotinib, fradaxin, zalutumab, tolituzumab, lidocaine, matuzumab, oxitizumab, panitumumab, PD168393, lapatinib, vandetanib, ridapepimut, canertinib, HuMAX-EGFR and CimaVax-EGF;

a HER2 inhibitor selected from the group consisting of ado-trastuzumab emtansine, trastuzumab, and pertuzumab; and

an immunostimulatory therapeutic agent selected from vidapentan, varliumab, monelizumab, KAHR-102, BGB324, enoblizumab, lirilumab, bazedoximab, pidilizumab, BL-8040, GDC-0919, IGN-311, erlotinzumab, bornauzumab, samalizumab, plerixafor, ganitumab, pexodartinib, trabedersen and galuninisertib.

18. The combination of claim 17, wherein the second compound is selected from erlotinib, palbociclumab, nivolumab, atuzumab, ipilimumab, avizumab, dovacizumab, trastuzumab, cetuximab, pertuzumab, and panitumumab.

19. The combination of claim 18, wherein the second compound is erlotinib, cetuximab, trastuzumab, or pertuzumab.

20. The combination of claim 18, wherein the second compound is palivizumab, nivolumab, atuzumab, ipilimumab, dulvacizumab, avizumab.

21. A pharmaceutical composition comprising a therapeutically effective amount of the combination of claim 1 and at least one pharmaceutically acceptable excipient.

22. The pharmaceutical composition of claim 21, wherein the compound of formula (I) is selected from:

(S) -6, 8-di-trideuteromethyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -6-bromo-8-trideuteromethyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -8-pentadeuterated ethyl-6- (trifluoromethoxy) -2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -6-chloro-8-trideuteromethyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -6-chloro-5, 7-di-trideuteromethyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -6-bromo-5, 7-di-trideuteromethyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -6-trideuteromethyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -8-chloro-6-trideuteromethyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -8-Trideuteromethyl-6- (pentafluorosulfanyl) -2- (trifluoromethyl) -2H-chromene-3-carboxylic acid, and

(S) -8-trideuteromethyl-6- (trifluoromethoxy) -2- (trifluoromethyl) -2H-chromene-3-carboxylic acid; and

wherein the second compound is selected from:

PD-1 inhibitors selected from the group consisting of nivolumab, pidilizumab, pabollizumab, AMP-224, AMP-514, STI-A1110, TSR-043, AMP-514, and AUNP-12;

a PD-L1 inhibitor selected from RG 7446, BMS-936559, MSB0010718C, STI-A1010, avizumab, atelizumab and covaptumab;

a CTLA-4 inhibitor selected from ipilimumab or tremelimumab;

an OX-40 agonist selected from anti-OX 40, TIM3 antibody and Immuture IMP 701;

a CD137 agonist selected from urelumab and utomicilumab;

LAG-3 inhibitor BMS-986016;

an IDO inhibitor selected from GDC-0919, indoximod, 1-methyl-D-tryptophan, NLG919, epacadostat, and norharpagne;

a bispecific protein selected from ALT-801 and MEDI-565;

an EGFR inhibitor selected from butitabine, gefitinib, erlotinib, neratinib, afatinib, dacomitinib, cetuximab, erlotinib, fradaxin, zalutumab, tolituzumab, lidocaine, matuzumab, oxitizumab, panitumumab, PD168393, lapatinib, vandetanib, ridapepimut, canertinib, HuMAX-EGFR and CimaVax-EGF;

a HER2 inhibitor selected from the group consisting of ado-trastuzumab emtansine, trastuzumab, and pertuzumab; and

an immunostimulatory therapeutic agent selected from vidapentan, varliumab, monelizumab, KAHR-102, BGB324, enoblizumab, lirilumab, bazedoximab, pidilizumab, BL-8040, GDC-0919, IGN-311, erlotinzumab, bornauzumab, samalizumab, plerixafor, ganitumab, pexodartinib, trabedersen and galuninisertib.

23. The pharmaceutical composition of claim 22, wherein the compound of formula (I) is (S) -6-bromo-8-trideuteromethyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid and the second compound is erlotinib, palbociclumab or astuzumab.

24. The pharmaceutical composition of claim 21, wherein the compound of formula (I) is selected from:

(S) -6, 8-dichloro-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -6-chloro-8-methyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -6-bromo-8-methyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -6, 8-dimethyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -8-methyl-6- (trifluoromethoxy) -2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -8-ethyl-6- (trifluoromethoxy) -2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -6-chloro-5, 7-dimethyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -7- (tert-butyl) -6-chloro-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -6-pentafluorosulfanyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid,

(S) -6-pentafluorosulfanyl-8-methyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid, and

(S) -6-pentafluorosulfanyl-8-ethyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid;

and the second compound is selected from:

PD-1 inhibitors selected from the group consisting of nivolumab, pidilizumab, pabollizumab, AMP-224, AMP-514, STI-A1110, TSR-043, AMP-514, and AUNP-12; this section provides background information related to the present disclosure, which is not necessarily prior art.

A PD-L1 inhibitor selected from RG 7446, BMS-936559, MSB0010718C, STI-A1010, avizumab, atlizumab and dulvacizumab;

a CTLA-4 inhibitor selected from ipilimumab or tremelimumab;

an OX-40 agonist selected from anti-OX 40, TIM3 antibody and Immuture IMP 701;

a CD137 agonist selected from urelumab and utomicilumab;

LAG-3 inhibitor BMS-986016;

an IDO inhibitor selected from GDC-0919, indoximod, 1-methyl-D-tryptophan, NLG919, epacadostat, and norharpagne;

a bispecific protein selected from ALT-801 and MEDI-565;

an EGFR inhibitor selected from butitabine, gefitinib, erlotinib, neratinib, afatinib, dacomitinib, cetuximab, erlotinib, fradaxin, zalutumab, tolituzumab, lidocaine, matuzumab, oxitizumab, panitumumab, PD168393, lapatinib, vandetanib, ridapepimut, canertinib, HuMAX-EGFR and CimaVax-EGF;

a HER2 inhibitor selected from the group consisting of ado-trastuzumab emtansine, trastuzumab, and pertuzumab; and

an immunostimulatory therapeutic agent selected from vidapentan, varliumab, monelizumab, KAHR-102, BGB324, enoblizumab, lirilumab, bazedoximab, pidilizumab, BL-8040, GDC-0919, IGN-311, erlotinzumab, bornauzumab, samalizumab, plerixafor, ganitumab, pexodartinib, trabedersen and galuninisertib.

25. A method of treating cancer, comprising:

administering to an individual in need thereof a therapeutically effective amount of a combination according to claim 1.

26. The method of claim 25, wherein the combination comprises a therapeutically effective amount of the compound of formula (I) that causes a reduction of PGE-M in urine of at least 70%.

27. The method of claim 25, further comprising determining the expression of PD-L1 in the cancer of the individual.

28. The method of claim 25, further comprising determining the level of urinary PGE-M levels in the individual.

29. The method of claim 28, wherein the individual has colorectal cancer, non-small cell lung cancer, breast cancer, gastric cancer, pancreatic cancer, prostate cancer, or head and neck squamous cell carcinoma.

30. The method of claim 25, wherein the cancer is selected from melanoma, non-small cell lung cancer, colorectal cancer, head and neck cancer, renal cell carcinoma, urothelial cancer, merkel cell carcinoma, pancreatic cancer, breast cancer, gastric cancer, intestinal cancer, endometrial cancer, hepatobiliary cancer, urinary tract cancer, brain cancer, skin cancer, glioblastoma, prostate cancer, and ovarian cancer.

31. The method of claim 30, wherein the dose of the compound of formula (I) is from about 0.1 mg/kg/day to about 100 mg/kg/day and the dose of the second compound is from about 0.01 mg/kg/day to about 250 mg/kg/day.

32. The method of claim 31, wherein the dose of the second compound is reduced relative to when the second compound is administered alone.

33. The method of claim 25, wherein the compound of formula (I) is (S) -6-bromo-8-trideuteromethyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid, the second compound is erlotinib, and the cancer is lung cancer.

34. The method of claim 25, wherein the compound of formula (I) is (S) -6-bromo-8-trideuteromethyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid, the second compound is pabollizumab, and the cancer is colorectal cancer.

35. The method of claim 25, wherein the compound of formula (I) is (S) -6-bromo-8-trideuteromethyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid, the second compound is pabollizumab, and the cancer is melanoma.

36. The method of claim 25, wherein the compound of formula (I) is (S) -6-bromo-8-trideuteromethyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid, the second compound is atezumab, and the cancer is colorectal cancer.

37. The method of claim 25, wherein the compound of formula (I) is (S) -6-bromo-8-trideuteromethyl-2- (trifluoromethyl) -2H-chromene-3-carboxylic acid, the second compound is atlizumab, and the cancer is lung cancer.